EPA/635/R-14/352
Preliminary Materials
www. ep a. gov/ir is
Preliminary Materials for the Integrated Risk Information System (IRIS)
Toxicological Review of Butyl Benzyl Phthalate (BBP)
(CASRN 85-68-7)
September 2014
NOTICE
This document is comprised of preliminary materials. This information is distributed solely for
the purpose of pre-dissemination review under applicable information quality guidelines. It has
not been formally disseminated by EPA. It does not represent and should not be construed to
represent any Agency determination or policy. It is being circulated for review of its technical
accuracy and science policy implications.
National Center for Environmental Assessment
Office of Research and Development
U.S. Environmental Protection Agency
Washington, DC
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
DISCLAIMER
This document is comprised of preliminary materials for review purposes only. This
information is distributed solely for the purpose of pre-dissemination review under applicable
information quality guidelines. It has not been formally disseminated by EPA. It does not represent
and should not be construed to represent any Agency determination or policy. Mention of trade
names or commercial products does not constitute endorsement or recommendation for use.
This document is a draft for review purposes only and does not constitute Agency policy.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
CONTENTS
PREFACE ix
1. INTRODUCTION 1-1
1.1.BBP IN THE ENVIRONMENT 1-1
1.1.1. Production and Use 1-1
1.1.2. Environmental Fate 1-1
1.1.3. Human Exposure Pathways 1-2
1.2.SCOPE OF THE ASSESSMENT 1-3
2. METHODS FOR IDENTIFYING AND SELECTING STUDIES 2-1
2.1.DRAFT LITERATURE SEARCH AND SCREENING STRATEGY 2-1
2.2.SELECTION OF CRITICAL STUDIES IN EARLY STAGES OF DRAFT DEVELOPMENT 2-16
2.2.1. General Approach 2-16
2.2.2. Exclusion of Studies 2-17
2.3.STUDY CHARACTERISTICS THAT WILL BE CONSIDERED IN THE FUTURE EVALUATION AND
SYNTHESIS OF THE CRITICAL EPIDEMIOLOGICAL STUDIES FOR BBP 2-18
2.4.STUDY CHARACTERISTICS THAT WILL BE CONSIDERED IN THE FUTURE EVALUATION AND
SYNTHESIS OF THE CRITICAL EXPERIMENTAL STUDIES FOR BBP 2-33
3. PRELIMINARY EVIDENCE TABLES AND EXPOSURE-RESPONSE ARRAYS 3-1
3.1. DATA EXTRACTION FOR EPIDEMIOLOGICAL AND EXPERIMENTAL STUDIES: PREPARATION
OF PRELIMINARY EVIDENCE TABLES 3-1
3.2.EPIDEMIOLOGICAL STUDIES 3-2
3.2.1. Sexual Differentiation Measures 3-2
3.2.2. Male Reproductive Effects in Humans 3-6
3.2.3. Male Pubertal Development in Humans 3-9
3.2.4. Semen Parameters and Infertility 3-11
3.2.5. Female Reproductive Effects in Humans 3-16
3.2.6. Female Pubertal Development in Humans 3-17
3.2.7. Gynecological Conditions in Humans 3-20
3.2.8. Pregnancy Related Outcomes 3-24
3.2.9. Immune Effects in Humans 3-28
3.2,10.Thyroid Effects in Humans 3-40
3.2,11.Pulmonary Function in Humans 3-42
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
3,2,12.Neurodevelopmental Effects in Humans 3-43
3.2,13.Obesity Effects in Humans 3-47
3,2,14.Diabetes Effects in Humans 3-52
3.2,15.Cardiovascular Effects in Humans 3-56
3,2,16.Cancer Effects in Humans 3-58
3.3.EXPERIMENTAL STUDIES 3-59
3.3.1. Male Reproductive Effects 3-59
3.3.2. Female Reproductive Effects 3-82
3.3.3. Developmental Effects 3-107
3.3.4. Liver Effects 3-125
3.3.5. Kidney Effects 3-139
3.3.6. Pancreatic Effects 3-156
3.3.7. Hematopoietic Effects 3-162
3.3.8. Thyroid Effects 3-179
3.3.9. Immune Effects 3-186
3.3,lO.Neurological Effects 3-188
3.3,11.Other Toxicity Effects 3-189
3,3,12.BBP Metabolite Studies 3-196
3.4. PRELIMINARY MECHANISTIC INFORMATION FOR BBP 3-215
4. REFERENCES 4-1
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
TABLES
Table 2-1. Database search strategy for BBP 2-2
Table 2-2. Summary of additional search strategies for BBP 2-2
Table 2-3. Inclusion criteria used to identify animal studies of health-related endpoints,
supporting data, or secondary literature 2-9
Table 2-4. Summary of search terms: targeted epidemiology search 2-10
Table 2-5. Inclusion criteria used to identify epidemiology studies of health-related endpoints 2-11
Table 2-6. Summary of additional search strategies for epidemiology studies of phthalate
exposure in relation to health-related endpoints 2-13
Table 2-7. Primary source epidemiological studies examining health effects of BBP 2-13
Table 2-8. General and outcome-specific considerations for BBP study evaluation 2-30
Table 2-9. Questions and relevant experimental information for the evaluation of experimental
animal studies 2-34
Table 3-1. Evidence pertaining to BBP and sexual differentiation effects in humans 3-2
Table 3-2. Evidence pertaining to BBP and reproductive hormones in adult men 3-6
Table 3-3. Evidence pertaining to BBP and the timing of male puberty or sex hormones in boys 3-9
Table 3-4. Evidence pertaining to BBP and semen parameters or infertility in adult men or
couples 3-11
Table 3-5. Evidence pertaining to BBP and reproductive hormones in adult women 3-16
Table 3-6. Evidence pertaining to BBP and timing of female puberty or sex hormones in girls 3-17
Table 3-7. Evidence pertaining to BBP and gynecological conditions in humans 3-20
Table 3-8. Evidence pertaining to BBP and pregnancy outcomes in humans 3-24
Table 3-9. Evidence pertaining to BBP and allergy/immune effects in humans 3-28
Table 3-10. Evidence pertaining to BBP and asthma/wheezing and hypersensitivity in humans 3-35
Table 3-11. Evidence pertaining to BBP and thyroid hormones in humans 3-40
Table 3-12. Evidence pertaining to BBP and pulmonary function in humans 3-42
Table 3-13. Evidence pertaining to BBP and neurodevelopmental effects in humans 3-43
Table 3-14. Evidence pertaining to BBP and obesity in humans 3-47
Table 3-15. Evidence pertaining to BBP and diabetes/insulin resistance in humans 3-52
Table 3-16. Evidence pertaining to BBP and cardiovascular disease risk factors in humans 3-56
Table 3-17. Evidence pertaining to BBP and cancer in humans 3-58
Table 3-18. Evidence pertaining to male reproductive puberty effects and indicators of
reproductive development following oral exposure to BBP 3-59
Table 3-19. Evidence pertaining to male reproductive toxicity following oral exposure to BBP:
Alterations in hormone concentrations, mating, and sperm decrements 3-64
Table 3-20. Evidence pertaining to male reproductive toxicity following oral exposure to BBP:
Histopathological changes and malformations in adults and offspring 3-69
Table 3-21. Evidence pertaining to male reproductive toxicity following oral exposure to BBP:
Decrease in androgen-dependent tissue weights 3-75
Table 3-22. Evidence pertaining to female reproductive toxicity following oral exposure to BBP 3-82
Table 3-23. Evidence pertaining to pregnancy outcomes following oral exposure to BBP:
Measures of embryotoxicity 3-97
Table 3-24. Evidence pertaining to developmental effects following oral exposure to BBP:
Teratogenicity 3-107
Table 3-25. Evidence pertaining to developmental effects following oral exposure to BBP:
offspring body weight 3-116
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Table 3-26. Evidence pertaining to liver effects in animals following oral and inhalation exposure
toBBP 3-125
Table 3-27. Evidence pertaining to kidney effects in animals following oral and inhalation
exposure to BBP 3-139
Table 3-28. Evidence pertaining to pancreatic effects in animals following oral and inhalation
exposure to BBP 3-156
Table 3-29. Evidence pertaining to hematopoietic effects in animals following oral and
inhalation exposure to BBP 3-162
Table 3-30. Evidence pertaining to thyroid effects in animals following oral exposure to BBP 3-179
Table 3-31. Evidence pertaining to immune effects in animals following oral exposure to BBP 3-186
Table 3-32. Evidence pertaining to neurological effects in animals following oral exposure to
BBP 3-188
Table 3-33. Evidence pertaining to other toxicity effects in animals following oral exposure to
BBP 3-189
Table 3-34. Evidence pertaining to toxicity effects in animals following exposure to BBP
metabolites 3-196
Table 3-35. Summary of mechanistic endpoints evaluated following BBP administration 3-216
FIGURES
Figure 1-1. Chemical structure of BBP 1-1
Figure 2-1. Literature search approach for BBP 2-8
Figure 3-1. Exposure-response array of male reproductive puberty effects and indicators of
reproductive development following oral exposure to BBP 3-63
Figure 3-2. Exposure-response array of male reproductive toxicity following oral exposure to
BBP: alterations in hormone concentrations, mating, and sperm decrements 3-68
Figure 3-3. Exposure-response array of male reproductive toxicity following oral exposure to
BBP: external and internal malformations 3-74
Figure 3-4. Exposure-response array of male reproductive toxicity following oral exposure to
BBP: decrease in androgen-dependent tissue weights 3-81
Figure 3-5. Exposure response array of female reproductive toxicity following oral exposure to
BBP: weights and pregnancy outcomes 3-95
Figure 3-6. Exposure response array of other female reproductive parameters following oral
exposure to BBP 3-96
Figure 3-7. Exposure-response array of pregnancy outcomes following oral exposure to BBP 3-105
Figure 3-8. Exposure-response array of fetal measures following oral exposure to BBP 3-106
Figure 3-9. Exposure-response array of developmental effects following oral exposure to BBP:
teratogenicity 3-114
Figure 3-10. Exposure-response array of developmental effects following oral exposure to BBP:
malformations 3-115
Figure 3-11. Exposure-response array of developmental effects following oral exposure to BBP:
fetal body weight 3-123
Figure 3-12. Exposure-response array of developmental effects following oral exposure to BBP:
pup weight 3-124
Figure 3-13. Exposure-response array of liver weight effects following oral exposure to BBP 3-137
Figure 3-14. Exposure-response array of liver histopathological effects following oral exposure
to BBP 3-138
Figure 3-15. Exposure-response array of kidney weight effects following oral exposure to BBP 3-154
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Figure 3-16. Exposure-response array of kidney histopathological effects following oral
exposure to BBP 3-155
Figure 3-17. Exposure-response array of pancreatic effects following oral exposure to BBP 3-161
Figure 3-18. Exposure-response array of hematopoietic effects following oral exposure to BBP:
spleen and thymus weights 3-177
Figure 3-19. Exposure-response array of hematopoietic histopathological effects following oral
exposure to BBP 3-178
Figure 3-20. Exposure-response array of thyroid effects following oral exposure to BBP 3-185
Figure 3-21. Exposure response array of other health effects following oral exposure to BBP 3-195
Figure 3-22. Summary of in vivo or in vitro mechanistic data by mechanistic category following
oral exposure to BBP 3-217
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
ABBREVIATIONS
ADME absorption, distribution, metabolism, MBP
and excretion MBzP
AGO anogenital distance MCPP
ALT alanine aminotransferase MDI
ANOVA analysis of variance MECPP
B B P butyl benzyl phthalate
BMI body mass index MEHHP
BP blood pressure
BPA bisphenolA MEHP
BW body weight MEOHP
CASRN Chemical Abstracts Service Registry MEP
Number MGH
CCCEH Columbia Center for Children's MIBP
Environmental Health M M E F
CERHR Center for the Evaluation of Risks to MMP
Human Reproduction MOA
CI confidence interval MW
Con A Concanavalin A NCEA
DBF dibutyl phthalate
DEP di-ethyl phthalate NHANES
DEHP di(2-ethylhexyl)phthalate
DHEAS dehydroepiandrosterone NHS
DIBP diisobutyl phthalate NIOSH
DINP diisononyl phthalate
DNA deoxyribonucleic acid NRC
DPP dipentyl phthalate NTP
EPA Environmental Protection Agency OR
FEVi forced expiratory volume in 1 second ORD
FSH follicle stimulating hormone PAH
FVC forced vital capacity PCB
GD gestational day PCO
E2 estradiol PCOS
feNO fractional exhaled nitric oxide PDI
GGT gamma glutamyl transferase PEF
HOMA homeostatic model assessment PND
HOMA-IR homeostatic model assessment of PNW
insulin resistance PPS
HERO Health and Environmental Research PVC
Online RfD
HOME Health Outcomes and Measures of the SD
Environment SE
IgE immunoglobulin E SFF
ICC intra-class correlation coefficient SHBG
IL interleukin T3
IRIS Integrated Risk Information System T4
IQR interquartile range TSCATS
ISAAC International Study of Asthma and
Allergies in Children TSH
LABC levator ani bulbocavernosus VOC
LH luteinizing hormone WHO
LOD level of detection
LOQ level of quantification
m-RNA messenger ribonucleic acid
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monobutyl phthalate
monobenzyl phthalate
mono-(3-carboxypropyl) phthalate
mental delay index
mono(2-ethyl-5-carboxypentyl)
phthalate
mono-(2-ethyl-5-
hydroxyhexyl)phthalate
mono-(2-ethylhexyl) phthalate
mono-(2-ethyl-5-oxohexyl) phthalate
monoethyl phthalate
Massachusetts General Hospital
monoisobuyl phthalate
maximal midexpiratory flow
monomethyl phthalate
mode of action
molecular weight
National Center for Environmental
Assessment
National Health and Nutrition
Examination Survey
Nurses Health Study
National Institute for Occupational
Safety and Health
National Research Council
National Toxicology Program
odds ratio
Office of Research and Development
polycyclic aromatic hydrocarbon
polychlorinated biphenyl
polycystic ovarian morphology
polycystic ovarian syndrome
psychomotor delay index
peak expiratory flow
postnatal day
postnatal week
preputial separation
polyvinyl chloride
reference dose
standard deviation
standard error
Study for Future Families
sex-hormone binding globulin
triiodothyronine
thyroxine
Toxic Substances Control Act Test
Submissions
thyroid stimulating hormone
volatile organic compound
World Health Organization
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
2 PREFACE
3 This draft document presents preliminary materials for an assessment of butyl benzyl
4 phthalate (BBP) prepared by the United States Environmental Protection Agency's (EPA) Integrated
5 Risk Information System (IRIS) Program. These preliminary materials include a planning and
6 scoping summary, information on the approaches used to identify pertinent literature, results of the
7 literature search, approaches for selection of studies for hazard identification, presentation of
8 critical studies in evidence tables and exposure-response arrays, and mechanistic information for
9 BBP. This material is being released for public review and comment prior to a public meeting,
10 providing an opportunity for the IRIS Program to engage in early discussions with stakeholders and
11 the public on data that may be used to identify adverse health effects and characterize dose-
12 response relationships.
13 The planning and scoping summary includes information on the uses of BBP, occurrence of
14 BBP in the environment, and the rationale and scope for the development of the assessment This
15 information is responsive to recommendations in the 2009 National Research Council (NRC) report
16 Science and Decisions: Advancing Risk Assessment (NRC, 2009) related to planning and scoping in
17 the risk assessment process.
18 The preliminary materials are also responsive to the NRC 2011 report Review of the
19 Environmental Protection Agency's Draft IRIS Assessment of Formaldehyde fNRC. 20111 The IRIS
20 Program's implementation of the NRC recommendations is following a phased approach that is
21 consistent with the NRC's "Roadmap for Revision" as described in Chapter 7 of the formaldehyde
22 review report The NRC stated that "the committee recognizes that the changes suggested would
23 involve a multi-year process and extensive effort by the staff at the National Center for
24 Environmental Assessment and input and review by the EPA Science Advisory Board and others."
25 Phase 1 of implementation has focused on a subset of the short-term recommendations, such as
26 editing and streamlining documents, increasing transparency and clarity, and using more tables,
27 figures, and appendices to present information and data in assessments. Phase 1 also focused on
28 assessments near the end of the development process and close to final posting. Phase 2 of
29 implementation is focused on assessments that are in the beginning stages of assessment
30 development The IRIS BBP assessment is in Phase 2 and represents a significant advancement in
31 implementing the NRC recommendations. In the development of this assessment many of the
32 recommendations are being implemented in full, while others are being implemented in part.
33 Achieving full and robust implementation of certain recommendations will be an evolving process
34 with input and feedback from the public, stakeholders, and independent external peer review.
35 Phase 3 of implementation will incorporate the longer-term recommendations made by the NRC,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 including the development of a standardized approach to describe the strength of evidence for
2 noncancer effects.
3 In May 2014, the NRC released their report reviewing the IRIS assessment development
4 process. As part of this review, the NRC reviewed current methods for evidence-based reviews and
5 made several recommendations with respect to integrating scientific evidence for chemical hazard
6 and dose-response assessments. In their report, the NRC states that EPA should continue to
7 improve its evidence-integration process incrementally and enhance the transparency of its
8 process. The committee did not offer a preference but suggests that EPA consider which approach
9 best fits its plans for the IRIS process. The NRC recommendations will inform the IRIS Program's
10 efforts in this area going forward. This effort is included in Phase 3 of EPA's implementation plan.
11 The literature search strategy, which describes the processes for identifying scientific
12 literature, screening studies for consideration, and identifying primary sources of health effects
13 data, is responsive to NRC recommendations regarding the development of a systematic and
14 transparent approach for identifying the primary literature for analysis. The preliminary materials
15 also describe EPA's approach for the selection of critical studies to be included in the evidence
16 tables, as well as the approach for evaluating methodological features of studies that will be
17 considered in the overall evaluation and synthesis of evidence for each health effect. The
18 development of these materials is in response to the NRC recommendation to thoroughly evaluate
19 critical studies with standardized approaches that are formulated and based on the type of research
20 (e.g., observational epidemiology or animal bioassays). In addition, NRC recommendations for
21 standardized presentation of key study data are addressed by the development of the preliminary
22 evidence tables and preliminary exposure-response arrays for primary health effect information.
23 EPA welcomes all comments on the preliminary materials in this document, including the
24 following:
25 • the clarity and transparency of the materials;
26 • the approach for identifying pertinent studies;
27 • the selection of critical studies for data extraction to preliminary evidence tables and
28 exposure-response arrays;
29 • any methodological considerations that could affect the interpretation of or confidence in
30 study results; and
31 • any additional studies published or nearing publication that may provide data for the
32 evaluation of human health hazard or dose-response relationships
33 The preliminary evidence tables and exposure-response arrays should be regarded solely as
34 representing the data on each endpoint that have been identified as a result of the draft literature
35 search strategy. They do not reflect any conclusions as to hazard identification or dose-response
36 assessment.
37 After obtaining public input and conducting additional study evaluation and data
38 integration, EPA will revise these materials to support the hazard identification and dose-response
39 assessment in a draft Toxicological Review that will be made available for public comment
This document is a draft for review purposes only and does not constitute Agency policy.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
2 1. INTRODUCTION
3 This introduction contains a planning and scoping summary for the Integrated Risk
4 Information System (IRIS) assessment of butyl benzyl phthalate (BBP). The planning and scoping
5 summary includes information on the properties, sources, and uses of BBP, occurrence and fate of
6 BBP in the environment, potential for human exposure, and the rationale for the development of
7 this assessment
8 1.1. BBP IN THE ENVIRONMENT
9 1.1.1. Production and Use
10 BBP (Chemical Abstract Service Registry Number [CASRN] 85-68-7) is a plasticizer used in a
11 wide range of materials including polyvinyl chloride (PVC)-based flooring, other plastics, adhesives,
12 coatings for automobiles, polyvinyl and cellulose resins, organic intermediates, sealants, foams,
13 inks, car care products, and cosmetics (HSDB, 2009). Between 50 and 100 million pounds were
14 imported or manufactured in United States in 2012 (http://www.epa.gov/oppt/cdr/index.html).
15
16
17 Figure 1-1. Chemical structure of BBP (HSDB. 2009).
18 1.1.2. Environmental Fate
19 If released to air, BBP will exist in both the vapor and particulate phases in the atmosphere.
20 Vapor-phase BBP will be photolytically degraded with a half-life of about 1.5 days. Particulate-
21 phase BBP will be removed from the atmosphere by wet or dry deposition. Once in soil, BBP is
22 tightly absorbed given a high organic carbon partition coefficient (Koc). Binding to soil organic
23 material limits volatilization as a route of dissipation. Biodegradation in aerobic soil and water is
24 expected to occur over days or weeks. Anaerobic biodegradation rates are expected to be slower. If
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 released into water, BBP is expected to adsorb to suspended solids and sediment Measured
2 bioconcentration factors of 9.4-772 suggest that concentrations in aquatic organisms may vary, but
3 metabolism of the chemical diminishes the likelihood of accumulation [HSDB, 2009]. As noted by
4 Wormuthetal. [2006], the majority of phthalates that are found in the environment come from
5 slow release from plastics and other phthalate-containing articles. Certain waste streams, sludges,
6 and contaminated sites, however, may contain higher levels of phthalates than other sites.
7 1.1.3. Human Exposure Pathways
8 The manner that humans are exposed to phthalates, along with the magnitude of exposures,
9 has changed over time as the quantities and uses of phthalates have changed. Human exposure to
10 phthalates occurs mainly in occupational or household settings because they are used and released
11 from products in the home environment. Environmental concentrations of phthalates are typically
12 the highest in house dust and they may be present in food due to the use of phthalates in packaging
13 and food preparation materials. For most phthalates, food ingestion is the dominant pathway of
14 exposure, with dust exposures (ingestion and dermal contact] and inhalation also being important
15 in some circumstances. Infant and toddler exposures occur due to teething and playing with plastic
16 toys that contain phthalates [Wormuthetal.. 2006].
17 The presence of phthalates or their metabolites in a body matrix, such as blood or urine,
18 provides evidence of exposure to that chemical. The predominant metabolite of BBP in humans is
19 monobenzyl phthalate [MBzP]. Zota etal. [2014] evaluated the prevalence and temporal trends of
20 MBzP in urine samples collected as part of the National Health and Nutrition Examination Survey
21 [NHANES] conducted between 2001 and 2010. MBzP was found in more than 98% of the urine
22 samples for each time period, and MBzP levels decreased recently, starting at about 10.4 ng/mL in
23 the 2001-2002 cycle and dropping to about 7.0 ng/mL in the 2009-2010 cycle.
24 Intake exposures can be estimated on a pathway-basis by combining exposure media
25 concentrations and contact rates. Using this approach, Clark etal. [2011] determined a median
26 intake of BBP of between 0.5 and 1.5 |ig/kg-day for various lifestages as defined by the authors:
27 adults (20-70 years of age], teens (12-19 years of age], children (5-11 years of age], toddlers
28 (0.5-4 years of age], and infants (0-0.5 years of age]. Toddlers had the highest intake noted.
29 Pathways the authors assessed include ingestion of food, drinking water, dust/soil, and inhalation
30 of air. Ingestion of food accounted for more than half of the total exposure for all age groups except
31 infants, with the remainder primarily due to incidental ingestion of dust and a minor contribution
32 due to inhalation of indoor air. For both the formula- and breast-fed infants, ingestion of dust
33 accounted for approximately 94% of exposure, with ingestion of food comprising most of the
34 remainder. Ingestion of food represented approximately 60% of total exposure for the adults and
35 inhalation of spray paints comprised most of the remainder in the estimates by Wormuth et al.
36 [2006]. who determined total intakes of <0.5 [ig/kg-day, except for infants and toddlers, who had
37 intakes between 0.5 and 1.0 |ig/kg-day.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 Wittassek et al. [2011] reported median intakes of BBP in the range of 0.1-0.9 ug/kg-day based
2 on a literature survey or urinary biomonitoring data and intake estimates provided therein. Their
3 review included U.S. estimates generated using data from the NHANES 2001-2002 cycle to
4 ascertain exposures in the range of 0.7-0.9 ug/kg-day. Qianetal. [2014] used NHANES 2007-2008
5 data and found a median intake of 0.3 ug/kg-day and a 95th percentile intake of 1.7 ug/kg-day.
6 Christensen et al. [2014] combined the data from NHANES 2005-2008 and found similar results to
7 Qianetal. [2014]. with a median over that time span of 0.2 ug/kg-day and a 95th percentile intake
8 of 1.0 ug/kg-day.
9 1.2. SCOPE OF THE ASSESSMENT
10 The National Research Council has recommended that, "cumulative risk assessment based on
11 common adverse outcomes is a feasible and physiologically relevant approach to the evaluation of
12 the multiplicity of human exposures and directly reflects EPA's mission to protect human health"
13 [NRG, 2008, pll]. They envisioned facilitating the process by "defining the groups of agents that
14 should be included for a given outcome" [NRG, 2008, p!2]. In humans, the NRC cited results from
15 the NHANES that demonstrate exposure to multiple phthalates in most people [NRC, 2008, p23-
16 25]. Recent reports on human exposure to phthalates suggest that the indoor environment is
17 thought to contribute to over 60% of BBP exposure in children [CHAP. 2014. Appendix El. p. 35]
18 and 94% of exposure in infants [Clark etal.. 2011]. The unique exposure scenarios and potential
19 sensitivities of children contribute to the need for an assessment of phthalate toxicity. This IRIS
20 assessment will help to inform EPA programs and regions of the potentially unique vulnerabilities
21 of children to BBP exposure and enable future cumulative risk assessments that assess effects on
22 human health outcomes that might be associated with BBP and other phthalates. EPA's previous
23 IRIS assessment of BBP [U.S. EPA. 1993] included an oral reference dose (RfD) and qualitative
24 cancer assessment (classified as Group C, a possible human carcinogen]. Since that time, a number
25 of experimental animal and epidemiological studies have been published for BBP.
This document is a draft for review purposes only and does not constitute Agency policy.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
2 2.METHODS FOR IDENTIFYING AND SELECTING
3 STUDIES
4 The National Research Council [NRG. 2011] recommended that the U.S. Environmental
5 Protection Agency (EPA) develop a detailed search strategy utilizing a graphical display
6 documenting how initial search findings are narrowed to the final studies that are selected for
7 further evaluation on the basis of inclusion and exclusion criteria. Following these
8 recommendations, a literature search and screening strategy was applied to identify literature
9 related to characterizing the health effects of butyl benzyl phthalate (BBP). This strategy consisted
10 of a search of online scientific databases and other sources, casting a wide net in order to identify all
11 potentially pertinent studies. In subsequent steps, references were screened to exclude papers not
12 pertinent to an assessment of the health effects of BBP, and remaining references were sorted into
13 categories for further evaluation. Section 2.1 describes the literature search and screening strategy
14 in detail. The NRC [NRG. 2011] further recommended that after studies are identified for review by
15 utilizing a transparent search strategy, the next step is to summarize the details and findings of the
16 most pertinent studies in the evidence tables. The NRC suggested that such tables should provide a
17 link to the references, and include details of the study population, methods, and key findings. This
18 approach provides for a systematic and concise presentation of the evidence. The NRC also
19 recommended that the methods and findings should then be evaluated with a standardized
20 approach. The approach that was outlined identified standard issues for the evaluation of
21 epidemiological and experimental animal studies. Section 2.2 describes the approach taken for BBP
22 for selecting studies to be included in the preliminary evidence tables and exposure-response
23 arrays. Section 3 presents the selected studies in preliminary evidence tables and exposure-
24 response arrays, arranged by health effect.
25 2.1. DRAFT LITERATURE SEARCH AND SCREENING STRATEGY
26 The literature search for BBP was conducted in four online scientific databases, including
27 PubMed, Toxline, Web of Science, and the Toxic Substances Control Act Test Submissions (TSCATS]
28 database, in December 2012; the search was repeated in August 2013 and in April 2014. This
29 document is complete through April 2014. Additional updates will be performed at regular (e.g., 6-
30 month] intervals. The detailed search approach, including the search strings is presented in
31 Table 2-1. The search strings and search terms described for BBP captured studies using the parent
32 compound and metabolites. This search of online databases identified 1,105 citations (after
33 electronically eliminating duplicates]. The computerized database searches were also
34 supplemented by a manual search of citations from other regulatory documents (Table 2-2];
This document is a draft for review purposes only and does not constitute Agency policy.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
2
63 citations were obtained using these additional search strategies. In total, 1,166 citations were
identified using online scientific databases and additional search strategies.
Table 2-1. Database search strategy for BBP
Database
(search date)
Keywords3
PubMed
04/2014
08/2013
12/2012
"1-butyl 2-(phenylmethyl) ester l,2-Benzenedicarboxylicacid"[tw] OR "benzyl butyl ester
Phthalic acid"[tw] OR "Benzyl butyl phthalate"[tw] OR "Benzyl butylphthalate"[tw] OR "Benzyl
n-butyl phthalate"[tw] OR "Butyl benzyl phthalate"[tw] OR "Butyl phenylmethyl 1,2-
benzenedicarboxylate"[tw] OR "butyl phenylmethyl ester 1,2-Benzenedicarboxylic acid"[tw]
OR "Butylbenzyl phthalate"[tw] OR "n-Butyl benzyl phthalate"[tw] OR "Palatinol BB"[tw] OR
"Santicizer 160"[tw] OR "Sicol"[tw] OR "Unimoll BB"[tw] OR «"BBP"[tw] OR BzBP[tw]) AND
(phthalic OR phthalate OR phthalates))
Web of Science
04/2014
08/2013
12/2012
TS="l-butyl 2-(phenylmethyl) ester 1,2-Benzenedicarboxylic acid" OR TS="benzyl butyl ester
Phthalic acid" ORTS="Benzyl butyl phthalate" ORTS="Benzyl butylphthalate" ORTS="Benzyl
n-butyl phthalate" ORTS="Butyl benzyl phthalate" ORTS="Butyl phenylmethyl 1,2-
benzenedicarboxylate" ORTS="butyl phenylmethyl ester 1,2-Benzenedicarboxylic acid" OR
TS="Butylbenzyl phthalate" ORTS="n-Butyl benzyl phthalate" ORTS="Palatinol BB" OR
TS="Santicizer 160" OR TS="Sicol" OR TS="Unimoll BB" OR «TS="BBP" OR TS="BzBP") AND
(TS="phthalic" OR TS=phthalate*))
Toxline
04/2014
08/2013
12/2012
@OR+("l-butyl 2-(phenylmethyl) ester 1,2-Benzenedicarboxylic acid'Vbenzyl butyl ester
Phthalic acid"+"Benzyl butyl phthalate"+"Benzyl butylphthalate"+"Benzyl n-butyl
phthalate"+"Butyl benzyl phthalate"+"Butyl phenylmethyl l,2-benzenedicarboxylate"+"butyl
phenylmethyl ester 1,2-Benzenedicarboxylic acid'VButylbenzyl phthalate"+"n-Butyl benzyl
phthalate"+"Palatinol BB'VSanticizer 160"+"Sicol"+"Unimoll BB"+@term+@rn+85-68-7)
+@NOT+@org+pubmed+pubdart+crisp+tscats
TSCATS2
08/2013
85-68-7
4
5
6
7
aThe search strings and search terms described above captured studies using the parent compound and
metabolites.
Table 2-2. Summary of additional search strategies for BBP
Approach
used
Manual search
from reviews
conducted by
other
international
and federal
agencies
Source(s)
CPSC (2010). Toxicity review of butyl benzyl phthalate
(BBP).
ECJRC (2007). European Union risk assessment report
butyl benzyl phthalate.
Date
performed
06/2013
06/2013
Number of
additional
citations identified
1 citation
33 citations
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Electronic
forward
Search
through Web
of Science
Electronic
backward
Search
through Web
of Science
References
obtained
during the
assessment
process
Aso et al. (2005). A two generation reproductive toxicitv
study of butyl benzyl phthalatein rats. The Journal of
Toxicological Sciences, 30, 39-58.
Tyl et al. (2004). Reproductive toxicitv evaluation of
dietary butyl benzyl phthalate (BBP) in rats. Reproductive
Toxicology, 18, 241-264.
Nagao et al. (2000). Effect of butyl benzyl phthalate in
Sprague-Dawley rats after gavage administration: a two-
generation reproductive study. Reprod Toxicol 14(6): 513-
532.
Aso et al. (2005). A two generation reproductive toxicitv
study of butyl benzyl phthalatein rats. The Journal of
Toxicological Sciences, 30, 39-58.
Tyl et al. (2004). Reproductive toxicitv evaluation of
dietary butyl benzyl phthalate (BBP) in rats. Reproductive
Toxicology, 18, 241-264.
Nagao et al. (2000). Effect of butyl benzyl phthalate in
Sprague-Dawley rats after gavage administration: a two-
generation reproductive study. Reprod Toxicol 14(6): 513-
532.
BBP references obtained from submissions, full study
reports from HERO, or in previous assessment
06/2013
06/2013
06/2013
06/2013
06/2013
06/2013
08/2014
0 citations
0 citations
1 citation
0 citations
4 citations
3 citations
63 citations
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Background
Check
Searched a combination of CASRNs and synonyms on the
following databases:
ACGIH (http://www.acgih.org/home.htm)
ATSDR (http://www.atsdr.cdc.gov/substances/index.asp
CalEPA Office of Environmental Health Hazard Assessment
(http://www.oehha.ca.gov/risk.html)
OEHHAToxicity Criteria Database
(http://www.oehha.ca.gov/tcdb/index.asp)
Biomonitoring California-Priority Chemicals
(http://www.oehha.ca.gov/multimedia/biomon/pdf/Priori
tyChemsCurrent.pdf)
Biomonitoring California-Designated Chemicals
(http://www.oehha.ca.gov/multimedia/biomon/pdf/Desig
natedChemCurrent.pdf)
Cal/Ecotox Database
(http://www.oehha.ca.gov/scripts/cal ecotox/CHEMLIST.
ASP)
OEHHA Fact Sheets
(http://www.oehha.ca.gov/public info/facts/index.html)
Non-cancer health effects Table (RELs) and Cancer
Potency Factors (Appendix A and Appendix B)
(http://www.oehha.ca.gov/air/hot spots/index. html)
CPSC
(http://www.cpsc.gov)
eChemPortal
(http://www.echemportal.0rg/echemportal/participant/p
age.action?pagelD=9)
Environment Canada - Search entire site
(http://www.ec.gc. ca/default.asp?lang=En&n=ECD35C36)
Toxic Substances Managed Under CEPA
(http://www.ec.gc.ca/toxiques-
toxics/Default.asp?lang=En&n=98E80CC6-l)
Final Assessments (http://www.ec.gc.ca/lcpe-
cepa/default.asp?lang=En&xml=09F567A7-BlEE-lFEE-
73DB-8AE6C1EB7658)
Draft Assessments (http://www.ec.gc.ca/lcpe-
cepa/default.asp?lang=En&xml=6892C255-5597-C162-
95FC-4B905320F8C9)
EPA Acute Exposure Guideline Levels
(http://www.epa.gov/oppt/aegl/pubs/chemlist.htm)
EPA - IRISTrack/New Assessments and Reviews
(http://cfpub.epa.gov/ncea/iristrac/) to find dates
(http://www.epa.gov/ncea/iris/index.html) to find data
EPANSCEP
(http://www.epa.gov/ncepihom/)
EPA RfD/RfC and CRAVE meeting notes
EPA Science Inventory
(http://cfpub.epa.gov/si/)
(http://www.fda.gov/)
12/2012
7 citations added
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Federal Docket
(www.regulations.gov)
Health Canada First Priority List Assessments
(http://www.hc-sc.gc.ca/ewh-
semt/pubs/contaminants/psll-lspl/index-eng.php)
Health Canada Second Priority List Assessments
(http://www.hc-sc.gc.ca/ewh-
semt/pubs/contaminants/psl2-lsp2/index-eng.php)
IARC
(http://monographs.iarc.fr/htdig/search.html)
HER (TERA database)
(http://iter.ctcnet.net/publicurl/pub search list.cfm)
NAP -Search Site
(http://www.nap.edu/)
NCI
(http://www.cancer.gov)
NCTR
(http://www.fda.gov/AboutFDA/CentersOffices/OC/Office
ofScientificandMedicalPrograms/NCTR/default.htm)
National Institute for Environmental Health Sciences
(NIEHS)
http://www.niehs.nih.gov/
NICNAS (PEC only covered by eChemPortal)
(http://www.nicnas.gov.au/industry/aics/search.asp)
NIOSH
(http://www.cdc.gov/niosh/topics/)
NIOSHTIC2
(http://www2a.cdc.gov/nioshtic-2/)
NTP - RoC, status, results, and management reports
(http://ntpsearch.niehs.nih.gov/query.html)
OS HA
(http://www.osha.gov/dts/chemicalsampling/toc/toc che
msamp.html)
RTECS
http://www.ccohs.ca/search.html
(http://www.fda.gov/)
Federal Docket
(www.regulations.gov)
Health Canada First Priority List Assessments
(http://www.hc-sc.gc.ca/ewh-
semt/pubs/contaminants/psll-lspl/index-eng.php)
Health Canada Second Priority List Assessments
(http://www.hc-sc.gc.ca/ewh-
semt/pubs/contaminants/ps!2-lsp2/index-eng.php)
IARC
(http://monographs.iarc.fr/htdig/search.html)
HER (TERA database)
(http://iter.ctcnet.net/publicurl/pub search list.cfm)
This document is a draft for review purposes only and does not constitute Agency policy.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
NAP -Search Site
(http://www.nap.edu/)
NCI
(http://www.cancer.gov)
NCTR
(http://www.fda.gov/AboutFDA/CentersOffices/OC/Office
ofScientificandMedicalProRrams/NCTR/default.htm)
National Institute for Environmental Health Sciences
(NIEHS)
http://www.niehs.nih.gov/
NICNAS (PEC only covered by eChemPortal)
(http://www.nicnas.Rov.au/industry/aics/search.asp)
NIOSH
(http://www.cdc.Rov/niosh/topics/)
NIOSHTIC2
(http://www2a.cdc.Rov/nioshtic-2/)
NTP - RoC, status, results, and management reports
(http://ntpsearch.niehs.nih.Rov/query.html)
OS HA
(http://www.osha.Rov/dts/chemicalsamplinR/toc/toc che
msamp.html)
RTECS
http://www.ccohs.ca/search.html
FDA
(http://www.fda.Rov/)
Federal Docket
(www.reRulations.Rov)
Health Canada First Priority List Assessments
(http://www.hc-sc.RC.ca/ewh-
semt/pubs/contaminants/psll-lspl/index-enR.php)
Health Canada Second Priority List Assessments
(http://www.hc-sc.RC.ca/ewh-
semt/pubs/contaminants/psl2-lsp2/index-enR.php)
IARC
(http://monoRraphs.iarc.fr/htdiR/search.html)
HER (TERA database)
(http://iter.ctcnet.net/publicurl/pub search list.cfm)
NAP -Search Site
(http://www.nap.edu/)
NCI
(http://www.cancer.Rov)
NCTR
(http://www.fda.Rov/AboutFDA/CentersOffices/OC/Office
ofScientificandMedicalProRrams/NCTR/default.htm)
National Institute for Environmental Health Sciences
(NIEHS)
http://www.niehs.nih.Rov/
NICNAS (PEC only covered by eChemPortal)
(http://www.nicnas.Rov.au/industry/aics/search.asp)
NIOSH
(http://www.cdc.Rov/niosh/topics/)
This document is a draft for review purposes only and does not constitute Agency policy.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
NIOSHTIC2
(http://www2a.cdc.gov/nioshtic-2/)
NTP - RoC, status, results, and management reports
(http://ntpsearch.niehs.nih.gov/querv.html)
OS HA
(http://www.osha.gov/dts/chemicalsampling/toc/toc che
msamp.html)
RTECS
http://www.ccohs.ca/search.html
1
2
3
4
5
6
7
These citations were screened using the title, abstract, and in limited instances, full text for
pertinence to examining the health effects of BBP exposure. The citations were then screened using
inclusion criteria (Table 2-3) describing specific information to help identify primary source health
effect data, mechanistic and/or genotoxic data, as well as resources useful in preparation of the BBP
package. The process for screening the literature is described below and is shown graphically in
Figure 2-1.
8 • 99 references were identified as animal studies with health effects data and were
9 considered for data extraction to evidence tables and exposure-response arrays.
10 • 122 references were identified as supporting studies; of these, 12 were toxicokinetic
11 studies and 110 were mechanistic and genotoxicity studies.
12 • 142 references were identified as secondary literature (e.g., reviews and editorials, risk
13 assessments, and regulatory documents); these references were kept as additional
14 resources for development of the Toxicological Review.
15 • 782 references were excluded because these studies did not include primary source
16 data evaluating BBP in relation to any kind of toxicity or health endpoint, and did not
17 provide either supporting information (e.g., toxicokinetic or mechanistic/genotoxic
18 data) or secondary literature information (see Figure 2-1 for and Table 2-3 for inclusion
19 categories and criteria).
20 Note that some studies were identified as belonging to multiple categories. As a result, the
21 total number of studies in a given category may be less than the sum of the individual studies listed
22 in subcategories.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Database Searches
(see Table 2-i for keywords and limitsl
Pubmed
n=414
Web of
Science
n=528
Toxline
(incl. TSCATSJ
n=585
(After duplicates removed electronicaityf
n=1105
Phthalates -Epidemiological
Studies Search
|see Table 2-4 for keywords and limitsl
Primary Source Human Data
N=185
>ee Table 2-5 for Inclusion Criteria)
Additional Search
Strategies
(see Tasle 2-2 for
tr en 3d: and results!
Combined Dataset
(After all duplicates removed!
n=l,166
Manual Screening For Pertinence
{Title/Abstract/Full Text)
Excluded: No Primary Data on Toxic
Effects jn=782)
S3 Abstract Only
13S ^ot Chemical Specific
19 Manufacture/use
86 Chemical treatment, disposes,
remediation
123 Measurement Methods
20 Miscellaneous
122 Ecosystem Effects
130 Exposure levels
71 Fate and Transport
15 Chemical/Physical properties
12 Use in sample prep or assay
13 mixtures only
Other Studies:
Studies with Supporting Data (n=122j
12 Toxicottinetic
110 Mechanistic and Genotoxicity Studies
Secondary Literature (n=142)
100 Reviews, editorials
31 Regulatory documents
12 Risk assessments
Animal Primary
Source Health
Effects Studies
(n=99)
(See Table 2-3 for Inclusion
Criteria)
Human Primary
Source Health
Effects Studies
(n=81)
(See Table 2-7 for a listmg
of BBP specific
epidemiologies! studies)
2 Note: Studies containing multiple information categories were sorted into multiple tags. For this reason, the
3 subcategory numbers do not always add up to the category total.
4
5
Figure 2-1. Literature search approach for BBP.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 Table 2-3. Inclusion criteria used to identify animal studies of health-related
2 endpoints, supporting data, or secondary literature
Inclusion criteria3
Did the study evaluate effects of BBP or its metabolites known to be formed in humans?
Did the study evaluate effects in a tissue (organ) or cells derived from a tissue (organ)?
Did the study evaluate cellular, biochemical or molecular effects relevant to any mode of action?
or
Does the study include information from other agencies, risk assessments, or reviews that would aid in
the development of a toxicological review of BBP?
3
4 alf the answer is "no" to any of these criteria questions, the study was placed under "Excluded: No Primary Data on
5 Toxic Effects"
6
7 Six foreign language studies reporting pertinent evidence for hazard characterization
8 and/or dose-response were identified. These studies by Agramunt et al. [2011]: Lietal. [2004]:
9 Timofievskaya et al. [19881: Timofievskaya et al. [19801: Tyrkiel et al. [20071: and Zhuang et al.
10 [2008] were tagged under "kept for possible further review" (not shown in figure]. A translation
11 was requested for the study by Zhuang etal. [2008] as it is one of the two available studies
12 reporting endpoints considered relevant to neurological effects. The remaining foreign language
13 studies report evidence for effects already described in English language publications and
14 captured in the BBP draft evidence tables. They will be considered individually for translation and
15 inclusion in evidence tables during development of the draft assessment of the available evidence
16 of BBP-induced health effects.
17 Sixteen human studies were also identified from the initial literature search using the
18 search strings presented in Table 2-1. However, work being done concurrently on the development
19 of other phthalate preliminary materials revealed that this set of BBP epidemiology studies was
20 incomplete. Epidemiology studies frequently examine multiple compounds (e.g., metabolites of
21 several different phthalates]. The indexing terms and abstracts may not include a comprehensive
22 list of all of the specific phthalates examined, resulting in the inappropriate exclusion of studies and
23 the potential for introduction of bias in the selection process. Specifically, "negative" studies (i.e.,
24 studies that did not demonstrate an association between exposure and disease] are potentially
25 more likely to be missed than "positive" studies. This issue did not arise in the search process for
26 experimental (animal toxicology] studies, for which the test compound is virtually always identified
27 through search terms or key word searches of abstracts.
28 Another issue encountered in the development of the search and screening process for the
29 phthalate epidemiology studies relates to the duplication of efforts involved in the development of
30 EPA's health assessments for several individual phthalates (e.g., BBP, dibutyl phthalate [DBF],
31 diisobutyl phthalate [DIBP], di(2-ethylhexyl]phthalate [DEHP], di-ethyl phthalate [DEP], diisononyl
This document is a draft for review purposes only and does not constitute Agency policy.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 phthalate [DINP], and dipentyl phthalate [DPP]). In contrast to animal toxicology studies, most of
2 the epidemiology studies examine more than one phthalate, resulting in considerable overlap in the
3 sets of studies identified using individual-phthalate search terms. Full text screening of the same
4 studies identified in multiple searches results in an inefficient use of resources.
5 For these reasons, EPA developed a process for identifying epidemiological studies
6 evaluating phthalates by performing a single broad search to create a listing of epidemiological
7 studies of all phthalates mentioned above, from which the selection of studies examining potential
8 health effects of an individual phthalate could be drawn. This list records each of the phthalates
9 included in the study, based on information in the methods section of the paper, and the outcome(s)
10 examined. This literature search for epidemiological studies examining phthalates in relation to
11 health-related endpoints (from which the BBP studies were drawn) was conducted in PubMed, Web
12 of Science, and ToxNet databases in June 2013, using keywords and limits described in Table 2-4;
13 the search was updated in December 2013 and in June 2014. For this search, "phthalate" (and
14 related terms) rather than names of specific phthalates was used as the foundation of the search,
15 along with terms designed specifically to identify epidemiological studies. These terms were based
16 on terms used in previously identified epidemiology studies of six different phthalates.
17 Table 2-4. Summary of search terms: targeted epidemiology search
Database,
search date
June 2013 search
PubMed
06/2013
No date restriction
Web of Science
06/2013
No date restriction
ToxNet
06/2013
No date restriction
Merged
Reference Set
December 2013
search
Terms
(phthalate OR phthalates OR phthalic acid) AND
(human OR case-control OR pregnancy OR cohort OR
workers OR children OR survey)
(TS="phthalic acid" ORTS="phthalate" OR
TS="phthalates") AND (TS="humans" ORTS="human"
ORTS="case-control" ORTS="pregnancy" OR
TS="cohort" ORTS="workers" ORTS="child" OR
TS="children" OR TS="survey")
(phthalate OR phthalates OR phthalic acid) AND
(human OR case-control OR pregnancy OR cohort OR
workers OR children OR survey)
Merged dataset, with duplicates eliminated through
electronic screen
Epidemiology articles meeting inclusion criteria
PubMed
Web of Science
ToxNet
Merged Reference Set
Additional epidemiology articles meeting inclusion
criteria
Hits
Imported: 2,505
After duplicates deleted: 2,482
Imported: 1,840
After duplicates deleted: 1,836
Imported: 2,505
After duplicates deleted: 2,426
4,127
127
155
249
114
350
22
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
June 2014
search
PubMed
Web of Science
ToxNet (was not searched because no articles have
been found solely through this source in all the
previous searches)
Merged Reference Set
Additional epidemiology articles meeting inclusion
criteria
184
409
0
494
24
1
2 More than 4,000 citations were identified through this search. These were then screened
3 using inclusion criteria describing specific population (i.e., human), exposure measures,
4 comparison, and health effects (Table 2-5). Note that other studies obtained in the search, for
5 example mechanistic and pharmacokinetic studies, are excluded from consideration with respect to
6 the specific objective of this search (i.e., identification of epidemiology studies), but could be
7 included in other steps in the assessment Duplicate citations of the same article were excluded and
8 articles written in a language other than English were retained for subsequent review. Earlier
9 analyses that are updated in a subsequent paper (e.g., with a larger sample size) are not included as
10 a primary paper, but may be used as background material regarding study methods.
11 Table 2-5. Inclusion criteria used to identify epidemiology studies of health-
12 related endpoints
Inclusion criteria
Is the study population humans?
and
Is exposure to one or more phthalate (parent compound or metabolite(s)a...
- measured in air, dust, or biological tissue?
- based on knowledge of industrial hygiene (occupational settings)?
- based on knowledge of specific contamination sites or accidental exposure?
and
Does the study compare a health effect in higher versus lower or no exposure?
and
Does the study include a measure of one or more primary health effect endpoints relating to...
- sexual differentiation measures (e.g., male genital malformations, anogenital distance, gender-related
play behavior)
- male reproductive effects (e.g., steroidal and gonadotropin hormone levels, measures of male-
mediated infertility)?
- female reproductive effects (e.g., steroidal and gonadotropin hormone levels, measures of female-
mediated infertility, gynecological conditions)?
- pregnancy outcomes (e.g., birth weight, gestation age)?
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Inclusion criteria
- puberty (male and female) (e.g., timing of development, precocious puberty, gynecomastia)?
- neurodevelopment (infants and children) (e.g., standardized tests of reflexes, behavior, and
intelligence)?
- thyroid effects (e.g., thyroid stimulating hormone and thyroid hormones, subclinical and clinical thyroid
disease)?
- immune system effects (e.g., asthma, allergies, IgE levels, skin prick tests)?
- pulmonary function (e.g., standardized test of lung volume, diffusing capacity)?
- neurological effects (adults) (e.g., peripheral neuropathy, vision or hearing or other sensory tests)?
- liver effects (e.g., cholestasis, biomarkers of liver function)?
- kidney effects (e.g., end stage renal disease, biomarkers of kidney function)?
- diabetes and measures of insulin resistance?
- obesity (and other measures of adiposity)?
- cardiovascular disease (cause-specific incidence or mortality)?
- cardiovascular risk factors (e.g., triglyceride and lipid levels, blood pressure or hypertension)?
- cancer (cause-specific incidence or mortality)?
or
Does the study include a measure of one or more secondary health effect endpoints (to be considered
within context of mechanistic evidence) relating to...
- oxidative stress?
- inflammation?
- gene expression?
1
2 aFor BBP, metabolite the primary metabolite of interest is MBzP (monobenzyl phthalate).
3
4 One hundred and seventy-three epidemiological studies examining one or more phthalate
5 in relation to one or more endpoints were identified by the searches conducted through June 2014
6 (127 in the initial search, 22 in the December 2013 update and 24 in the June 2014 update)
7 (Figure 2-1). Other strategies were also used to supplement this broad search for epidemiology
8 studies of phthalates, resulting in the identification of 12 additional publications (Table 2-6), for a
9 total of 185 epidemiological studies. From this set of all of the epidemiological studies examining
10 any phthalate, 81 studies analyzed one or more health effects in relation to a measure of BBP
11 (Table 2-7).
This document is a draft for review purposes only and does not constitute Agency policy.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
2
Table 2-6. Summary of additional search strategies for epidemiology studies
of phthalate exposure in relation to health-related endpoints
Approach used
Testing and refinement of search terms based on terms used for the
identified articles within each category
Review of references cited in the identified list of epidemiology studies
("backward" search)
Electronic forward search through Web of Science of 1-3 studies within
each health endpoint category (early studies within each category
generally selected to maximize potential for citation in subsequent
publications)3
Date
performed
June 2014
July 2014
July 2014
Number of additional
citations identified
6
1
5
3
4
5
6
7
8
9
10
aThe following studies were used to conduct the forward searches (Trasande et al. (2013b); James-Todd et al.
(2012); Lind and Lind (2011); Boas et al. (2010); Cho et al. (2010); Engel et al. (2010); Lopez-Carrillo et al. (2010);
Wolff etal. (2010); Adibi et al. (2009); Chouetal. (2009); Hatch et al. (2008); Wolff etal. (2008); Meeker et al.
(2007); Stahlhut et al. (2007); Mauser et al. (2006); Reddyet al. (2006a); Jonssonetal. (2005); Swan etal. (2005);
Bornehagetal. (2004); Hoppin etal. (2004); Aschengrau et al. (1998); Heineman etal. (1992); Nielsen etal.
(1989); Nielsen etal. (1985))
11
12
Table 2-7. Primary source epidemiological studies examining health effects of
BBP
Outcome category
Sexual differentiation measures
(Table 3-1)
Male reproductive (semen
parameters, infertility, and
hormones)
(Tables 3-2 and 3-3)
Male pubertal development
(Table 3-4)
Reference3
Linetal. (2011a)
Main etal. (2006)
Suzuki etal. (2012)
Swan (2008)
Swan et al. (2010)
Buck Louis etal. (2014)
Mauser etal. (2006)
Mauser etal. (2007)
Joensen etal. (2012)
Jonsson etal. (2005)
Jurewicz et al. (2013)
Kranvogl et al. (2014)
Liu etal. (2012)
Meeker et al. (2009a)
Mendiola etal. (2011)
Mendiola etal. (2012)
Toshimaetal. (2012)
Tranfo et al. (2012)
Wirth et al. (2008)
Ferguson et al. (2014b)
Mieritzetal. (2012)
Mouritsen etal. (2013b)
BBP measure
MBzP (maternal urine)
MBzP (breast milk)
MBzP (maternal urine)
MBzP (maternal urine)
MBzP (maternal urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (maternal urine)
MBzP (urine)
MBzP (urine)
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Outcome category
Female pubertal development
(Table 3-5)
Female reproductive (infertility,
hormones, gynecological conditions)
(Tables 3-6 and 3-7)
Pregnancy-related outcomes (fetal
growth, preterm birth, pregnancy
loss)
(Table 3-8)
Allergy (rhinitis, eczema)
(Table 3-9)
Asthma
(Table 3-10)
Pulmonary Function (Table 3-11)
Reference3
Chen et al. (2013)
Chouetal. (2009)
Frederiksen et al. (2012)
Hart et al. (2013)
Lomenicketal. (2010)
Mouritsen etal. (2013b)
Buck Louis etal. (2013)
Hart et al. (2013)
Huang etal. (2010)
Itoh et al. (2009)
Reddy et al. (2006a)
Reddy et al. (2006b)
Sathyanarayana et al. (2014)
Upson etal. (2013)
Weuve et al. (2010)
Ferguson et al. (2014c)
Ferguson et al. (2014a)
Huang etal. (2014b)
Meeker et al. (2009b)
Philippatetal. (2012)
Suzuki etal. (2010)
Toft et al. (2012)
Wolff etal. (2008)
Ait Bamai etal. (2014)
Bornehag et al. (2004)
Callesen et al. (2014b)
Callesen et al. (2014a)
Hoppinetal. (2013)
Hsu etal. (2012)
Just etal. (2012b)
Kanazawa et al. (2010)
Kolariketal. (2008)
Sun etal. (2009)
Wang et al. (2014)
Ait Bamai etal. (2014)
Bertelsen et al. (2013)
Bornehag et al. (2004)
Callesen et al. (2014b)
Callesen et al. (2014a)
Hoppinetal. (2013)
Hsu etal. (2012)
Just etal. (2012a)
Kolariketal. (2008)
Sun etal. (2009)
Cakmak etal. (2014)
Hoppin (2004)
BBP measure
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (maternal serum)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (maternal serum)
MBzP (urine)
BBP (plasma)
BBP (plasma)
MBzP (maternal urine)
MBzP (urine)
MBzP (urine)
MBzP (maternal urine)
MBzP (maternal urine)
BBP (cord blood)
MBzP (maternal urine)
MBzP (maternal urine)
MBzP (maternal urine)
MBzP (maternal urine)
MBzP (maternal urine)
BBP (dust)
BBP (dust)
MBzP (urine)
BBP (dust)
MBzP (urine)
BBP (dust), MBzP (urine)
MBzP (maternal urine)
BBP (dust)
BBP (dust)
BBP (dust)
MBzP (maternal urine)
BBP (dust)
MBzP (urine)
BBP (dust)
MBzP (urine)
BBP (dust)
MBzP (urine)
BBP (dust), MBzP (urine)
MBzP (urine)
BBP (dust)
BBP (dust)
MBzP (urine)
MBzP (urine)
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Outcome category
Neurodevelopment
(Table 3-12)
Thyroid
(Table 3-13)
Obesity
(Table 3-14)
Diabetes and insulin resistance
(Table 3-15)
Other cardiovascular disease risk
factors (Table 3-16)
Cancer
(Table 3-17)
Reference3
Braunetal. (2014)
Chopra et al. (2014)
Kobrosly et al. (2014)
Tellez-Roio et al. (2013)
Whyatt et al. (2012)
Boas et al. (2010)
Dirtu et al. (2013)
Huang et al. (2007)
Meeker et al. (2007)
Buseretal. (2014)
Dirtu et al. (2013)
Hart et al. (2013)
Hatch et al. (2008)
Song et al. (2014)
Stahlhutetal. (2007)
Svensson et al. (2011)
Teitelbaum et al. (2012)
Huang etal. (2014a)
James-Todd et al. (2012)
Svensson et al. (2011)
Stahlhutetal. (2007)
Sunetal. (2014)
Trasandeetal. (2013a)
Shiue (2014)
Trasandeetal. (2013b)
Aschengrau etal. (1998)
Lopez-Carrillo et al. (2010)
BBP measure
MBzP (maternal urine)
MBzP (urine)
MBzP (maternal urine)
MBzP (maternal urine)
MBzP (maternal urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (maternal serum)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
MBzP (urine)
Work history
MBzP (urine)
1
2
3
4
5
The literature for both epidemiological and animal studies will be regularly monitored for
the publication of new studies. The documentation and results for this supplementary search can
be found on the Health and Environmental Research On-line (HERO) website1
[http://hero.epa.gov/BBP] and [http://hero.epa.gov/phthalates-humanstudies].
iHERO is a database of scientific studies and other references used to develop EPA's risk assessments aimed
at understanding the health and environmental effects of pollutants and chemicals. It is developed and
managed in EPA's Office of Research and Development (ORD) by the National Center for Environmental
Assessment (NCEA). The database includes more than 1,400,000 scientific articles from the peer-reviewed
literature. New studies are added continuously to HERO.
Note: The HERO database will be regularly updated as additional references are identified during assessment
development. Therefore, the numbers of references (by tag) displayed on the HERO webpage for BBP may
not match the numbers of references identified in Figure 2-1 (current through September 2014).
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l 2.2. SELECTION OF CRITICAL STUDIES IN EARLY STAGES OF DRAFT
2 DEVELOPMENT
3 2.2.1. General Approach
4 Each study retained following the literature search and screen was evaluated for aspects of
5 design, conduct, or reporting that could affect the interpretation of results and the overall
6 contribution to the synthesis of evidence for determination of hazard potential. Much of the key
7 information for conducting this evaluation can generally be found in the study's methods section
8 and in how the study results are reported. Importantly, this evaluation does not consider study
9 results or, more specifically, the direction or magnitude of any reported effects. For example,
10 standard issues for evaluation of experimental animal data identified by the NRC and adopted in
11 this approach include consideration of the species and sex of animals studied, dosing information
12 (dose spacing, dose duration, and route of exposure), endpoints considered, and the relevance of
13 the endpoints to the human endpoints of concern. Similarly, observational epidemiologic studies in
14 this approach for evaluation should consider the following:
15 • Approach used to identify the study population and the potential for selection
16 bias
17 • Study population characteristics and the generalizability of findings to
18 other populations
19 • Approach used for exposure assessment and the potential for information
20 bias, whether differential (nonrandom) or nondifferential (random)
21 • Approach used for outcome identification and any potential bias
22 • Appropriateness of analytic methods used
23 • Potential for confounding to have influenced the findings
24 • Precision of estimates of effect
25 • Availability of an exposure metric that is used to model the severity of
26 adverse response associated with a gradient of exposures
27
28 To facilitate the evaluation outlined above, evidence tables are constructed that
29 systematically summarize the important information from each study in a standardized tabular
30 format as recommended by the NRC (NRC. 2011). In general, the evidence tables include all studies
31 that inform the overall synthesis of evidence for hazard potential. At this early stage of study
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1 evaluation, the goal is to be inclusive. Exclusion of studies may unnecessarily narrow subsequent
2 analyses by eliminating information that might later prove useful. Premature exclusion might also
3 give a false sense of the consistency of results across the database of studies by unknowingly
4 reducing the diversity of study results. However, there may be situations in which the initial review
5 of the available data will lead to a decision to focus on a particular set of health effects and to
6 exclude others from further evaluation.
7 2.2.2. Exclusion of Studies
8 After the literature search was manually screened for pertinence, studies were excluded if
9 fundamental flaws were identified in their design, conduct, or reporting. The BBP experimental
10 animal database consists of studies designed to examine repeat-dose oral toxicity (including
11 chronic, subchronic, and short-term duration studies) and endpoint-specific toxicities (including
12 reproductive and developmental toxicity). All studies involved administration of BBP via oral or
13 inhalation routes. Acute or short-term studies are generally less pertinent for characterizing health
14 hazards associated with chronic exposure; there are 25 acute and short-term studies that are not
15 summarized in the preliminary evidence tables. In addition, studies using atypical exposure routes
16 (e.g., intraperitoneal or subcutaneous exposure) (4 studies), and studies that used a single high
17 dose (6 studies) when other multi-dose studies with similar endpoints were available, were also
18 not included in the preliminary evidence tables. Nevertheless, these studies will still be evaluated
19 as possible sources of supporting health effects information during assessment development Two
20 studies were identified that involved administration of very low doses (<1 ppm) of BBP. Following
21 the recommendations of a NTP-CERHR (2003) review, these studies were not included in the
22 evidence tables due to: (1) lack of dose-response data; (2) lack of analytical data on levels of BBP
23 in drinking water; (3) failure of the original laboratory to duplicate their findings; and (4) inability
24 of other reputable laboratories to duplicate the findings. In addition, five studies were not
25 summarized in the preliminary evidence tables because they presented data previously published
26 in other studies that are included in the preliminary evidence tables; four studies were not
27 summarized due to co-administration of other chemicals at the time of dosing; and one study was
28 not included in the preliminary evidence tables due to the presence of a respiratory infection
29 reported in the control colony. The remaining studies are all sources of health effects data that may
30 be used in the assessment The studies summarized in the evidence tables are considered the
31 "critical" studies from which the study methods and results are presented in preliminary evidence
32 tables and exposure-response arrays (Section 3).
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l 2.3. STUDY CHARACTERISTICS THAT WILL BE CONSIDERED IN THE
2 FUTURE EVALUATION AND SYNTHESIS OF THE CRITICAL
3 EPIDEMIOLOGICAL STUDIES FOR BBP
4 Several considerations will be used in EPA's evaluation of epidemiological studies of human
5 health effects of BBP. These considerations include aspects of the study design affecting the
6 internal or external validity of the results (e.g., population characteristics and representativeness,
7 exposure and outcome measures, confounding, data analysis), focusing on specific types of bias
8 (e.g., selection bias; information bias due to exposure misclassification), and other considerations
9 that could otherwise influence or limit the interpretation of the data. A study is externally valid if
10 the study results for the study population can be extrapolated to external target populations. An
11 internally valid study is free from different types of biases, and is a prerequisite for generalizing
12 study results beyond the study population. These issues are outlined in the Integrated Risk
13 Information System (IRIS) Preamble, and are described below.
14 Study Population
15 Evaluation of study population characteristics (including key socio-demographic variables
16 and study inclusion criteria) can be used to evaluate external validity (i.e., generalizability) and to
17 facilitate comparison of results across different study populations. Some aspects of the selection
18 process may also affect the interval validity of a study, resulting in a biased effect estimate.
19 The general considerations for evaluating issues relating to the study population include
20 adequate documentation of participant recruitment, including eligibility criteria and participation
21 rates, missing data, and loss to follow-up. This information is used to evaluate internal study
22 validity related to selection bias. Different types of selection bias that may occur include the
23 healthy worker effect, differential loss to follow up, Berkson's bias (relating to selection of
24 participants in hospital-based, case-control studies), and participation bias. It is important to note
25 that low participation rates, or differences in participation rates between exposed and non-exposed
26 groups or between cases and controls, is not evidence of selection bias. Rather, selection bias arises
27 from a differential pattern of participation with respect to both the exposure and the outcome, i.e.,
28 patterns of participation that would result in a biased effect estimate. An example of differential
29 participation would be when people with high levels of exposure and the outcome of interest are
30 more likely to participate than people with low levels of exposure and the outcome.
31 The available BBP studies have generally examined metabolites from many different
32 phthalates within the context of research on environmental exposures. Most of these studies rely
33 on objective exposure measures (e.g., biomonitoring data), some of which are collected prior to
34 onset of the outcomes being examined (e.g., in the prospective pregnancy cohort studies). Study
35 participants generally do not have knowledge of the study hypothesis or their exposure to BBP and
36 thus, knowledge of exposure or exposure level is unlikely to result in differential participation with
37 respect to outcomes. These study features should minimize the potential for selection bias.
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1 However, EPA will consider the possibility that a particular concern about the specific sources of
2 BBP, in conjunction with knowledge of specific health outcomes, may motivate people to participate
3 in a study or to continue participation throughout a follow-up period. In the absence of evidence
4 that any of these scenarios is likely to occur in a study, EPA will not consider selection bias as a
5 limitation of a study.
6 Exposure Considerations
7 General considerations for evaluating exposure include: (1) identifying how exposure can
8 occur (e.g., exposure sources, routes and media); (2) determining appropriate critical exposure
9 period(s) for the outcomes under study; (3) evaluating variability in the exposure metrics of
10 interest (e.g., temporal and spatial variability for environmental measures or inter-individual
11 variability for biomonitoring data) that can impact different types of exposure metrics (e.g.,
12 cumulative, average, or peak exposure); (4) determining if an appropriate analytical methodology
13 was employed (e.g., choice of biological matrix, sampling protocol, quantification approach);
14 (5) evaluating the choice of exposure surrogate evaluated (e.g., constituent chemical or group/
15 mixture); and (6) evaluating the classification of individuals into exposure categories. These six
16 considerations help determine the accuracy and precision of the exposure estimates, and the
17 likelihood of measurement error with respect to the exposure metrics used. Nondifferential
18 misclassification of exposure categories, for example, can also result from measurement error and
19 is expected to predominantly result in attenuated effect estimates (Blair etal.. 2007).
20 Some common sources of exposure to BBP include polyvinyl chloride (PVC) flooring, food,
21 and food packaging material (Zota etal.. 2014) with the primary route of exposure occurring
22 through ingestion and some exposure occurring via inhalation and dermal routes (see
23 Section 1.1.3). Thus, exposure to BBP is typically from multiple sources, and occurs episodically on
24 a daily basis. Exposure to BBP may be decreasing; a recent study of the U.S. general population
25 found that urinary concentrations of the BBP metabolite, MBzP, have decreased somewhat over
26 time and were 32% lower in 2009-2010 compared to 2001-2002 fZota etal.. 2014).
27 Urine provides an integrated measure of phthalate exposure from all sources.
28 Measurement of BBP metabolites, rather than the parent compound, is preferred because the
29 parent compound is metabolized very quickly and does not provide an accurate measure of
30 exposure. The simple monoester metabolite, MBzP, is the most commonly measured BBP
31 metabolite in epidemiologic studies. MBzP accounts for an estimated 73% of the urinary excretion
32 of BBP (Anderson et al., 2001). This value is based on data from a 24-person (all adults) controlled
33 dosing study (Koch etal.. 2012). EPA considers the use of MBzP to be a good proxy for total BBP
34 exposure.
35 Although urine measures are most commonly used in epidemiological studies of phthalate
36 exposure, measures in serum, semen, and breast milk have also been used. Studies examining BBP
37 metabolites in breast milk or serum have generally reported low levels of detection. One study in
38 Taiwan reported that MBzP above the limit of detection was found in 10% of breast milk samples
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1 from 30 women and 10% of the corresponding 30 cord blood samples. The correlation between
2 MBzP in maternal urine and breast milk was -0.27 and for maternal urine and cord blood was
3 -0.09 (Pearson correlation of log-transformed levels) [Lin etal.. 201 Ib). Hogbergetal. [2008]
4 reported that few breast milk (3 out of 42) samples in a study in Sweden had detectable MBzP
5 concentrations. Another study conducted among 60 men ages 18-26 years found that 10% of
6 serum samples and 18.6% of seminal plasma samples had MBzP concentrations above the limit of
7 detection [Frederiksenetal.. 2010). Correlation coefficients between MBzP measured in urine and
8 these other samples were not calculated because the detection rates were low [Frederiksenetal..
9 2010). The lower detection rate in tissues other than urine reduces EPA's confidence in BBP
10 metabolite measures in these biological matrices.
11 Given their first-order kinetics with half-lives on the order of hours (~5-12 hours for
12 MBzP) [Koch and Angerer, 2007], urinary phthalate metabolite concentrations peak shortly after
13 exposure. Thus, for single-time exposure scenarios (rather than multi-source, multiple time
14 exposure scenarios), urine sampled during this time of peak concentration could lead to
15 overestimates of average daily intake, and conversely, measurements made after concentrations
16 have peaked and declined could lead to underestimates of intake. One study conducted among
17 139 pregnant women in Puerto Rico found that sampling time was not a significant predictor of
18 urinary MBzP concentrations; that is, there was little difference in MBzP levels for women whose
19 samples were collected in early morning, morning, early afternoon, or evening time periods
20 (geometric mean specific gravity adjusted MBzP 4.5, 3.9, 4.2, and 4.7, respectively, for these four
21 periods, p = 0.74) [Cantonwine etal.. 2014). Urinary measures of BBP metabolite concentrations in
22 epidemiological studies are generally conducted using spot urine samples (i.e., collected at time of a
23 clinic or study examination visit) rather than at a specified time (e.g., first morning void) or in
24 24-hour urine samples. Although the time of sample collection described above may affect the
25 accuracy of an estimated intake for a single individual, studies of other phthalates (e.g., DEHP) have
26 demonstrated that on a group level, spot urine samples provide a reasonable approximation of
27 concentrations that would have been observed using full-day urine samples [Christensen et al..
28 2014] and that a single spot sample was reliable in ranking subjects according to tertile of MBzP
29 [Teitelbaum etal.. 2008). Based on this information, EPA does not consider the reliance on spot
30 urine samples for exposure estimation (including ranking of individuals into different BBP
31 categories) to be a major limitation for epidemiological studies. However because of the potential
32 for greater inaccuracy of estimates in the "tails" of the distribution, EPA will include additional
33 considerations (e.g., discussion of analysis of residuals, outliers) when evaluating analyses based on
34 use of BBP metabolites as continuous measures.
35 Another potential limitation of measurement of BBP metabolites in urine is the
36 reproducibility of phthalate metabolite concentrations over time; that is, how well does a single
37 measure reflect the key exposure metric (average, peak) for the critical exposure window of
38 interest For many short-lived chemicals, considerable temporal variability in exposure level is
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1 expected, and thus, repeated measures in the critical exposure window are preferred over a single
2 measurement Reproducibility is usually evaluated with the intraclass correlation coefficient (ICC),
3 a measure of the 'between-individual' variance divided by the total variance (between and within
4 individuals). A higher ICC indicates greater reproducibility (i.e., lower within-person variance). An
5 ICC of 0.64 for MBzP was reported in a study of 25 Hmong women ages 19-51 years with samples
6 collected 2-4 weeks apart (Pecketal., 2010). For MBzP measures in 46 women ages 35-49 years, a
7 moderate correlation was seen over a period of 2 days (ICC of 0.34 unadjusted, 0.53 creatinine-
8 adjusted) (Hoppinetal.. 2002). Similar values were seen in two studies in men with longer
9 sampling periods (approximately 3 months): in 33 men ages 18-22 years, the ICCs for MBzP in spot
10 urine samples were 0.38 (unadjusted) and 0.39 (osmolality-adjusted) in (Frederiksen et al., 2013),
11 and in 11 men with up to 9 spot urine samples collected on 3 consecutive days in each of 3 monthly
12 cycles, the ICC was 0.43 (Hauser et al., 2004). In studies of reproducibility of measures during
13 pregnancy, Cantonwine etal. (2014) reported ICCs of 0.37 and 0.41 (unadjusted and specific-
14 gravity adjusted) when comparing urine samples taken at approximately 18, 22, and 26 weeks of
15 gestation. ICCs of 0.35 and 0.28, respectively, were seen before pregnancy and in early pregnancy
16 (Braunetal.. 2012). and an ICC of approximately 0.65 was seen over a 6-week period in the last
17 trimester (Adibietal., 2008). Among women participating in the Nurses' Health Study (NHS) (in
18 2000-2001 for NHS and in 1996-1999 for NHS II), the ICC for samples collected 1-3 years apart
19 was 0.33 for all samples, and was 0.31 for first-morning samples (Townsendetal., 2013). Data for
20 children are sparse, limiting the ability to examine this source of uncertainty in this population: one
21 study evaluated variability in children aged 6-10 years old over a 6-month period (Teitelbaum et
22 al.. 2008) and reported ICCs of 0.47 (unadjusted) and 0.62 (creatinine-adjusted). The available data
23 highlight the value of repeated exposure measures collected during the appropriate critical period
24 for the outcome(s) under study.
25 Based on these studies, however, EPA does not consider the use of a single measurement to
26 be a major limitation in studies in adults in which the measure of exposure is closely aligned with
27 the relevant window(s) of exposure, if known, for the effect under study. EPA has greater
28 uncertainty, however, about measurements taken outside of the relevant time window (e.g., several
29 years after diagnosis, or the difference between first and third trimesters of pregnancy), and about
30 measurements taken in children.
31 Some studies present analyses using a combined "high molecular weight" phthalate
32 measure based on the summation of DEHP metabolites and MBzP. Because MBzP does not
33 represent a major contributor to this summation, EPA has not included data from these studies in
34 the BBP evidence tables.
35 EPA will also consider the potential for differential misclassification of biomarker measures
36 of exposure, for example in situations in which a health outcome (e.g., diagnosis with diabetes or
37 cancer) could lead to a behavioral change that result in a change in BBP exposure. This type of
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1 scenario adds an additional challenge to the interpretation of the BBP metabolites as valid
2 measures of exposure in a relevant time window(s) with respect to disease development
3 The distribution of exposure will also be considered in evaluating individual studies and
4 when comparing results among groups of studies. One consideration is the contrast of exposure
5 levels (i.e., the difference between "high" and "low"): a study with a very narrow contrast may not
6 have sufficient variability to detect an effect that would be seen over a broader range. Another
7 consideration is the absolute level of exposure, as different effect estimates may be expected in
8 studies examining different exposure levels even if they had similar exposure contrasts.
9 Primary Outcome Measures
10 The general considerations for evaluating issues relating to accuracy, reliability, and
11 biological relevance of outcomes include adequate length of follow-up to evaluate the outcomes of
12 interest, and use of appropriate ascertainment methods to classify individuals with regard to the
13 outcome (e.g., high sensitivity and specificity). With respect to continuous measures, such as
14 hormone concentrations or semen parameters, EPA will consider, in addition to assessing whether
15 reported parameters are outside normal physiological range, evidence of smaller changes in the
16 distribution of a parameter that may represent an effect on a population level [e.g., as is the case for
17 early childhood exposure to lead and decrements in intelligence as measured by IQ (U.S. EPA,
18 2013).
19 Issues relating to the assessment of the specific primary health effects are discussed below
20 and summarized in Table 2-8 at the end of Section 2.3.
21 Sexual differentiation
22 Cryptorchidism and hypospadias are two disorders of the development of the male
23 reproductive system. Cryptorchidism, or undescended testes, can be present at birth (congenital
24 Cryptorchidism) or can occur later during infancy and childhood (acquired Cryptorchidism).
25 Surgical correction (orchiopexy) is recommended in cases of Cryptorchidism that do not resolve
26 during infancy because long-term complications include impaired sperm production and increased
27 risk of testicular cancer (Virtanen et al.. 2007). Retractile testes can move back and forth between
28 the scrotum and the abdomen; this condition usually resolves by puberty and is not associated with
29 reproductive or other complications. Classification criteria for Cryptorchidism that involve
30 testicular positioning are commonly used in clinical research (Tohn Radcliffe Hospital
31 Cryptorchidism Study Group, 1988: Scorer, 1964). EPA will consider the definition used and age
32 range in interpreting studies of Cryptorchidism or related outcomes.
33 In animal toxicology studies, anogenital distance (AGD) is a routine marker to assess
34 endocrine disruption; this marker has only recently been adapted for use in epidemiological
35 studies. One study in adult men reported associations between decreased AGD and measures
36 relating to infertility (Eisenbergetal., 2011): most studies have used this measure in infants,
37 however, as a marker of endocrine environment during development. It is important to consider
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1 general size, in addition to sex, in the evaluation of AGD, for example by incorporating birth weight
2 or length (e.g., calculation of "anogenital index" by dividing AGD by weight). With regard to
3 reproducibility of this measure, a low degree of between-observer variability was found using a
4 standardized protocol and trained observers [Romano-Riqueraetal., 2007: Salazar-Martinez etal.,
5 2004]. Because of the importance of size and age in the interpretation of this measure, EPA has
6 greater confidence in studies with measures taken at birth or over a narrow age range and lesser
7 confidence in studies among a group spanning a larger age range.
8 Gender-related behaviors, as measured by the Pre-School Activities Inventory [Golombok
9 and Rust. 1993] or other scales, have also been examined in relation to direct or indirect measures
10 of fetal testosterone levels, including studies of BBP. This outcome measure has been examined in
11 studies of relatively rare genetic conditions (e.g.., congenital adrenal hyperplasia and complete
12 androgen insensitivity syndrome], as well as studies focusing on the normal variability seen in the
13 general population (reviewed in Hines. 2006]. EPA will consider evidence pertaining to the
14 reliability and validity of the Pre-School Activities Inventory in its evaluation of studies using this
15 scale.
16 Male and female reproductive outcomes
17 The BBP literature includes studies of reproductive and gonadotropin hormone levels in
18 men and studies of semen parameters that can be indicative of reduced fertility. The details of the
19 laboratory procedures, including information on the basic methods, level of detection, and
20 coefficient of variation, are important considerations for hormone assays and measures of semen
21 parameters. The World Health Organization (WHO] laboratory methods for analysis of sperm
22 counts and semen parameters (see, for example. WHO. 1999] are generally recognized as standards
23 in this field. EPA will consider studies that reference these methods, regardless of which revision
24 used, to be reliable measures.
25 Much of the focus of the research on male steroidal and gonadotropin hormones in the BBP
26 database concerns testosterone. One issue with respect to these measures is the estimation method
27 used for free testosterone. Based on the analysis by Vermeulen et al. (1999]. EPA will consider
28 estimates based on total testosterone divided by immunoassay-derived sex-hormone binding
29 globulin (SHBG] levels to be most reliable.
30 The BBP literature also includes studies of reproductive hormones in women. In addition to
31 the general considerations regarding hormone assays noted above, timing within a menstrual cycle
32 for studies of pre- and peri-menopausal women and timing with respect to gestational age for
33 studies of women during pregnancy are also be an important considerations for interpretation of
34 reproductive hormone concentrations.
35 Other female reproductive outcomes included in the BBP literature include endometriosis.
36 Endometriosis can be symptomless, or can lead to surgical intervention; it is often diagnosed as
37 part of a work-up for infertility. Variability in clinical presentation and in access and use of health
38 care services present considerable challenges to conducting epidemiological studies of this
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1 condition [Holt and Weiss. 2000). Confirmation of "case" and "control" status (i.e., presence or
2 absence of endometriosis) by ultrasound or clinical evaluation is recommended to reduce outcome
3 misclassification, and representation of the source population should be carefully considered.
4 Infertility is generally defined clinically and for research purposes as the inability to
5 conceive a clinically-recognized pregnancy after 12 months of intercourse of regular frequency
6 without use of contraceptives. Fecundity or fecundability are terms for the capacity for
7 reproduction. "Time to pregnancy" (i.e., the number of cycles of unprotected intercourse before
8 conception) has been used as a measure of fecundability in studies of environmental and
9 occupational exposures (Bairdetal.. 1986: Baird and Wilcox. 1985). Time to pregnancy is a
10 measure of a couple's fecundability, incorporating effects that can be manifested through the male
11 or female (or both). Considerations in time to pregnancy studies include the source of data (i.e.,
12 retrospective or prospective designs), and incorporation of information on "non-pregnancy
13 planners" (Weinberg et al.. 1994).
14 Timing of male and female puberty, and conditions of unusual pubertal development
15 Pubertal development in humans is often assessed using timing of peak height velocity
16 ("growth spurt") and secondary markers of sexual development Secondary markers for females
17 include breast development (thelarche) and pubic hair development (pubarche), and age at first
18 period (menarche). Secondary markers for males include gonadal development (gonadarche) and
19 pubic hair development, and age at first sperm emission (spermarche).
20 Evaluation of breast, pubic hair, and gonadal development is frequently performed using
21 the Tanner stages (Marshall and Tanner. 1970.1969). which places the individual in one of five
22 stages, ranging from pre-pubertal (stage 1) to adult maturation (stage 5). However, the process of
23 this staging is not straightforward, and is most reliable when performed by trained personnel
24 (rather than by the individual or a parent, for example) (Slough etal., 2013: Schlossberger et al.,
25 1992: Espelandetal., 1990). Age at menarche is considered to more reliable when assessed via
26 self-report (Koprowskietal.. 2001). although reliability may decrease with increasing time since
27 menarche (Cooper etal.. 2006). Additionally, hormone levels may sometimes be used to evaluate
28 pubertal development Individuals may vary widely in the timing of these developmental
29 milestones.
30 Several clinical syndromes are known to disrupt the timing and order of markers of
31 pubertal development Considerations in the diagnosis of either precocious or delayed puberty
32 include the diagnostic criteria used and the source of the information (e.g., whether collected from
33 medical records or from self- or parental report). For females, precocious puberty is usually
34 defined as the onset of puberty before the age of 8 years, while delayed puberty is usually defined
35 as the lack of pubertal development by the age of 13 years (Marshall and Tanner. 1969):
36 corresponding ages in male are before the age of 9 years for precocious puberty and lack of
37 pubertal development by the age of 14 years for delayed puberty (Marshall and Tanner, 1970).
38 Clinical evaluation would involve hormone assays to distinguish between gonadotropin dependent
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1 ("central"), gonadotropin independent ("peripheral"), or a combination of both (Traggiai and
2 Stanhope. 2003) forms of these conditions.
3 Pregnancy-related outcomes
4 Infant birth weight and gestational age are two outcomes commonly used in reproductive
5 epidemiology studies. EPA considers analyses of the various indices for both outcomes (fetal
6 growth and gestational age) to be informative with respect to hazard identification, but will
7 consider each separately as they address different issues. Gestational duration can be measured as
8 a continuous outcome or dichotomous outcome such as preterm birth. Preterm births include
9 infants delivered earlier than 37 gestational weeks, and those delivered earlier than 32 gestational
10 weeks are classified as very preterm births. Different measures of fetal growth restriction are often
11 examined in epidemiological studies. In addition to the continuous measure of birth weight,
12 another commonly used measure of fetal growth restriction is the categorical variable of low birth
13 weight (defined as <2,500 g). Small for gestational age (defined as birth weight less than the 10th
14 percentile for the gestational birth weight distribution) is considered a better measure of fetal
15 growth rate as it takes into consideration gestational duration, and would be preferred over a
16 measure of birth weight in a study that includes preterm births. Birth weight and gestational
17 duration can also be examined as continuous variables, often in analysis that excludes preterm or
18 low birth weight births, so that the focus of the analysis is on variability within the "normal" range.
19 EPA considers birth weight obtained from medical records to be a reliable source as this is a
20 very accurate and precise measurement Although more prone to measurement error than birth
21 weight measures, gestational age can be estimated from several approaches. Some of these include
22 ultrasonography, estimates based on date of last menstrual period based on maternal recall, or
23 from clinical examination based on antenatal or newborn assessments (which may include an
24 ultrasound). Menstrual dating of gestational age dependent on maternal recall of the last menstrual
25 period can be subject to considerable measurement error in some cases, so ultrasonography-based
26 estimates may be considered more accurate (Savitzetal.. 2002: Taipale and Hiilesmaa. 2001).
27 Expectant mothers can encounter pregnancy loss either through a stillbirth (fetal death
28 after 20 gestational weeks) or from a spontaneous abortion also known as a miscarriage (fetal
29 death during the first 20 gestational weeks). Pregnancy loss can occur even before a clinically
30 recognized pregnancy; early pregnancy (or "subclinical") loss, determined by measurement of
31 human chorionic gonadotropin, is very common, accounting for approximately 20% of pregnancies
32 (Wilcoxetal., 1988). Thus, complete ascertainment of pregnancy loss requires this type of
33 monitoring for subclinical loss.
34 Immune-related outcomes: allergy and asthma
35 Skin prick testing is a standard method for assessing atopy (allergic disease) used in some
36 epidemiologic studies. Other studies use an assessment protocol based on reported history of
37 symptoms (e.g., rhinitis, hay fever) or specific types of allergies. These can be considered
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1 complementary types of measures: skin prick tests provide information on a defined set of
2 potential antigens to which a person may be exposed, and symptom-based evaluations provide
3 information on experiences of individuals and the variety of exposures they encounter. Studies
4 comparing questionnaire responses with skin prick tests in children have reported relatively high
5 specificity (89-96%) and positive predictive value (69-77%) for self-reported history of pollen or
6 pet dander allergy or for answers to a combination of questions incorporating itchy eyes with nasal
7 congestion in the absence of a cold or flu (Braun-Fahrlander etal.. 1997: Dotterudetal.. 1995). The
8 validity was somewhat lower for a more restricted set of questions (nasal congestion in the absence
9 of a cold or flu; specificity 83%, positive predictive value 52%) (Braun-Fahrlander etal.. 1997).
10 Based on these data, EPA considers allergy history based only on rhinitis symptoms to have a
11 greater likelihood of outcome misclassification compared with those based on a combination of
12 symptoms.
13 Epidemiologic studies of asthma typically use a questionnaire-based approach to define
14 asthma based on symptoms relating to wheezing episodes or shortness of breath, reported history
15 of asthma attacks, or use of asthma medication, usually for a period defined as "current" or in the
16 past year. Much of this work is based upon the American Thoracic Society questionnaire (Ferris.
17 1978) or subsequent instruments that built upon this work, including the International Society of
18 Arthritis and Allergies in Children Questionnaire and the European Community Respiratory Health
19 Survey. These questionnaire-based approaches have been found to have an adequate level of
20 specificity and positive predictive value for use in etiologic research (Ravault and Kauffmann. 2001:
21 Pekkanen and Pearce. 1999: BurneyetaL 1989: Burney and Chinn. 19871 EPA considers
22 outcomes defined over a recent time period (e.g., symptoms in the past 12 months) to be more
23 relevant within the context of concurrent exposure measurements compared with outcomes
24 defined over a lifetime (e.g., ever had asthma).
25 Pulmonary function
26 The American Thoracic Society has published guidelines for equipment performance
27 requirements, validation, quality control, test procedures, and reference equations for each type of
28 spirometric measurement (Miller etal.. 2005). as well as the interpretation of testing results
29 (Pellegrino etal.. 2005). Lung function varies by race or ethnic origin, gender, age, and height, and
30 is best compared when normalized to the expected lung function based on these variables
31 (Pellegrino etal., 2005: Hankinsonetal., 1999). Some measures (e.g., forced expiratory volume in
32 1 second [FEVi] and peak expiratory flow [PEF]) exhibit diurnal variation (Chan-Yeung, 2000:
33 Lebowitz etal.. 1997): thus, time of day of the lung function measures should also be considered.
34 Neurodevelopment
35 With respect to neurodevelopmental outcomes, a major consideration is the assessment
36 tool(s) used by the study investigators; details of the assessment method, or references providing
37 this information, should be provided. In addition, EPA also looks for discussion of (or reference to)
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1 validation studies and the appropriateness of the tool for evaluation in the specific study population
2 (e.g., age range, language).
3 Thyroid
4 Thyroid-related endpoints examined in epidemiological studies of BBP include thyroid
5 hormones (triiodothyronine, T3, and thyroxine, T4) and thyroid stimulating hormone (TSH) (or
6 thyrotropin) produced by the pituitary.
7 As with other hormone assays, the details of the laboratory procedures, including
8 information on the basic methods, limit of detection, and coefficient of variation, are important
9 considerations for the hormone assays. Thyroid hormones are generally measured in serum,
10 although they may also be measured in dried blood spots, such as are collected from newborn
11 infants in screening for congenital hypothyroidism. Studies in older age groups have also shown a
12 very high correlation (r = 0.99) between thyroid hormone levels measured in dried blood spots and
13 levels in serum fHofmanetal.. 20031
14 With respect to thyroid hormones, time of day and season of sampling are two main
15 potential sources of variability. For example, serum TSH measured shortly after midnight may be
16 as much as twice as high as the value measured in late afternoon (Brabant etal., 1991: Weeke and
17 Gundersen, 1978). The evidence with respect to seasonal variability is mixed (Plasqui etal., 2003:
18 Nicolau etal.. 1992: Simonietal.. 1990: Behalletal.. 1984: Postmes etal.. 1974) and this effect is
19 likely to be smaller than that of time of day. The impact of these sources of variation will depend on
20 whether they are also related to BBP (i.e., whether BBP levels vary diurnally or seasonally). If this
21 is the case, failure to address these factors in the design or analysis could result in confounding of
22 the observed association, with the direction of this bias determined by the direction of the
23 association between these factors and BBP. If this is not the case, the lack of consideration of time
24 of day or seasonality would result in greater variability in the hormone measures, and would thus
25 result in more imprecise (but not biased) estimates was located. EPA has not found studies
26 examining seasonal variation in BBP levels. With respect to variability relating to time of day, as
27 noted previously, one study of 139 pregnant women in Puerto Rico reported little variation by
28 sampling time (early morning, morning, early afternoon, or evening) of specific gravity-adjusted
29 MBzP (Cantonwine etal.. 2014)). Based on these data, EPA does not consider the lack of
30 consideration of time of day or season in the analysis of thyroid outcomes to be a likely source of
31 bias, but recognizes the limited nature of the available data.
32 Obesity
33 Most of the study of obesity measures in the BBP database are based on body mass index
34 (BMI, calculated as kg/m2) or waist circumference using measurements taken as part of the data
35 collection protocol. BMI is highly correlated with body fat, and standardized cut-points have been
36 established for characterization of "normal" (BMI between 18.5 and 24.9 kg/m2), "overweight"
37 (BMI between 25.0 and 29.9 kg/m2), and "obese" (BMI > 30.0 kg/m2) categories. Waist
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 circumference is also highly correlated with body fat, and is a more direct measure of abdominal
2 obesity. EPA notes that use of self-reported weight (e.g., report of pre-pregnancy weight) would
3 not be considered to be as reliable as actual measurements.
4 Diabetes and measure of insulin resistance
5 In the BBP database, diabetes has been assessed by a variety of biomarkers of glucose and
6 insulin and by self-report of diabetes diagnosis. Oral glucose tolerance testing and glycosolated
7 hemoglobin (HbAlc) are used clinically and in epidemiological research [Selvinetal.. 2011). Self-
8 report of prevalent diabetes can have high sensitivity and specificity in comparison to diagnosed
9 diabetes based on validated medical record data [Oksanenetal., 2010: Leikauf and Federman,
10 2009]. The biomarker-based classifications, however, offer an added advantage of being able to
11 include undiagnosed disease. EPA will consider these points in assessing the reliability and validity
12 of the diabetes measures used in the studies. None of the currently available studies assessed
13 diabetes through cause of death data; sensitivity of diabetes assessed using cause of death data is
14 low, even if underlying and other contributing cause of death fields are included [Cheng etal..
15 20081
16 Insulin resistance, a marker of diabetes risk, can be measured using the homeostatic model
17 assessment (HOMA) method, a physiologically-based structural model, using fasting glucose and
18 insulin or C-peptide concentrations. HOMA is a validated tool for the estimation of insulin
19 resistance in epidemiology studies, and requires a single measurement of fasting glucose and
20 insulin [Wallace etal.. 2004). Although the mean of three samples taken at 5-minute intervals
21 results in a more precise estimate, insulin resistance estimated using a single baseline
22 measurement is well correlated with that using the mean of three measurements when used to
23 estimate a group mean. Therefore, EPA does not consider the use of a single measurement as an
24 input to the HOMA model to be a limitation.
25 Cancer
26 With respect to studies of cancer, EPA considers the source of the outcome data (e.g., cause
27 of death data, hospital cancer registry data, hospital discharge data, histopathology reports) in its
28 evaluation of the accuracy of the data. An additional issue is the validity of mortality data as a
29 representation of cancer incidence; mortality data for cancer types with a high survival rate may
30 underrepresent disease incidence, require additional considerations with respect to determining
31 appropriate time windows of exposure, and may lead to biased risk estimates if survival is related
32 to exposure.
33 Confounding
34 The general considerations for evaluating issues relating to potential confounding include
35 consideration of which factors may be potential confounders (i.e., those that are strongly related to
36 both the exposure and the outcome under consideration, and are not intermediaries on a causal
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1 pathway), adequate control for these potential confounders in the study design or analysis, and
2 where appropriate, quantification of the potential impact of mismeasured or unmeasured
3 confounders. Uncontrolled confounding by factors that are positively associated with both the
4 exposure (e.g., BBP) and health endpoint of interest, and those that are inversely associated with
5 both exposure and health endpoint, will result in an upward bias of the effect estimate.
6 Confounding by factors that are positively associated with exposure and inversely associated with
7 the health endpoint (or vice versa) will result in a downward bias of the effect estimate.
8 Potential confounding by other phthalates
9 Few studies have reported results of analyses evaluating the correlation between MBzP and
10 metabolites of other phthalates. In an analysis conducted by EPA of 5,109 samples from the
11 2003-2008 National Health and Nutrition Examination Survey (NHANES) participants aged
12 >6 years, the pairwise Spearman correlation coefficient between MBzP and monoethyl phthalate
13 (MEP) (the primary metabolite of DEP) was low (0.28). A more moderate correlation was seen
14 between MBzP and DEHP metabolites (correlations of approximately 0.5); higher correlations were
15 seen with monoisubutyl phthalate (MIBP) (the primary metabolite of DIBP, Spearman r = 0.58) and
16 with MBP (the primary metabolite of DBF, Spearman r = 0.70). Similar or somewhat lower
17 correlations were seen between MBzP and other phthalate metabolites in 463 men seen in an
18 infertility clinic (Hauser et al., 2006), in 319 pregnancy women (Whyatt et al., 2012), and in
19 600 reproductive age women in a study of endometriosis (Buck Louis et al.. 2013). EPA will
20 evaluate the potential for confounding by examining the similarity of the results seen with different
21 metabolites. Thus, for example, lack of adjustment for MBzP would not be considered a limitation
22 in a study in which an association was seen with MBzP that was not seen with MBP; however, this
23 lack of adjustment would be considered a limitation if an association of similar or higher
24 magnitude was seen for both of metabolites.
25 Potential confounding by demographic factors
26 Age, race/ethnicity, and sex are considered important explanatory factors for most types of
27 outcomes measured in epidemiological research. In NHANES 2009-2010 data, urinary MBzP levels
28 decreased with age (geometric means of 15.1, 8.54, and 5.94 [ig/g-creatinine, respectively, in ages
29 6-11,12-19, and >20 years) (CDC, 2013). Smaller differences were seen when comparing
30 distributions by sex (geometric means of 6.21 and 7.29 |ig/g-creatinine, respectively, in males and
31 females), and by ethnicity (geometric means of 7.53, 6.83, and 6.50 |ig/g-creatinine, respectively, in
32 Mexican Americans, non-Hispanic whites, and non-Hispanic blacks). EPA will consider these
33 differences in assessing the potential influence of demographic factors on observed effect estimates
34 for BBP.
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1 Potential confounding by other factors
2 Some of the health effects under consideration may have strong associations with other risk
3 factors. For example, smoking is associated with increased risk of low birth weight and preterm
4 births, and with infertility. Abstinence time is strongly related to sperm concentration measures.
5 In evaluating the potential for confounding by any of these factors, EPA will review evidence
6 pertaining to the strength and direction of its association with BBP (or its metabolites).
7 Data Analysis
8 The general considerations for evaluating issues relating to data analysis include adequate
9 documentation of statistical assumptions and analytic approach (including addressing skewness of
10 exposure or outcome variable and shape of exposure-response), consideration of sample size and
11 statistical power, and use of appropriate statistical methods for the study design.
12 One other issue specific too much of the BBP literature concerns the optimal approach to
13 addressing urinary volume or dilution in the analysis of spot urine or first morning void samples.
14 Options include use of creatinine- or specific gravity-adjusted metabolite concentrations, or use of
15 unadjusted concentrations. Although use of some kind of correction factor has been advocated for
16 studies of obesity (Goodman etal., 2014), a simulation study reported that creatinine-adjusted
17 exposure measures may produce biased effect estimates for outcomes that are strongly related to
18 factors affecting creatinine levels, of which obesity is a prime example (Christensenetal., 2014).
19 EPA recognizes the lack of consensus at this time, as well as the need for continued research into
20 the potential bias introduced by different analytic approaches. Based on current understanding of
21 this issue, EPA prefers results using unadjusted concentrations for outcomes strongly related to
22 creatinine levels; for other outcomes, EPA does not have a basis for preferring one type of analysis
23 over another.
24 Table 2-8. General and outcome-specific considerations for BBP study
25 evaluation
General considerations
Study population
Study population and setting: geographic area, site, time period, age and sex
distribution, other details as needed (may include race/ethnicity,
socioeconomic status)
Recruitment process; exclusion and inclusion criteria, knowledge of study
hypothesis; knowledge of exposure and outcome
Participation rates: total eligible; participation at each stage and for final
analysis group and denominators used to make these calculations
Length of follow-up, loss to follow-up
Comparability: participant characteristic data by group, data on non-
participants
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Exposure
• Biological matrix or target tissue/organ (e.g., urine, serum, semen, breast
milk)
• Level of detection (LOD) or level of quantitation (LOQ)
• Exposure distribution (e.g., central tendency, interquartile range), proportion
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Relevant exposure
timewindow(s)
• Up to 6 mo preceding semen sample collection
Infertility
Measures
Consideration of
confounding
Relevant exposure
time window(s)
• Definition, source of data
• Age, smoking, alcohol use, heavy metal exposure, radiation time (consider if
these are related to exposure)
• Time preceding or during attempt to become pregnant
Timing of puberty
Measures
Consideration of
confounding
Relevant exposure
time window(s)
• Source of data (e.g., self-report, physician assessment)
• Age, sex, ethnicity, body size, nutritional status (consider if these are related
to exposure)
• In utero? Up to 12 mo preceding transition from one stage to another
stage?
Gestational age
Measures
Consideration of
confounding
Relevant exposure
time window(s)
• Source of data and estimation procedure (ultrasound; last menstrual period
or clinical assessment)
• Smoking, pregnancy complications, assisted reproduction technologies
(consider if these are related to exposure)
• In utero
Birth weight
Measures
Consideration of
confounding
Relevant exposure
time window(s)
• Source of data (e.g., medical records, birth certificate)
• Gestational age, maternal age, ethnicity, infections, pregnancy complications
(e.g., pre-eclampsia), nutritional intake, smoking, alcohol/drug use, weight
gain during pregnancy; maternal height/BMI, heavy metal exposures
(consider if these are related to exposure)
• In utero
Immune - allergy and
asthma
Measures
Consideration of
confounding
Relevant exposure
time window(s)
• Number of allergens used in skin prick testing or allergen-specific IgE assay;
sensitivity/specificity of specific questions used in history assessment
• Age, family history (consider if these are related to exposure)
• For current conditions (e.g., asthma in past 12 mo): up to 12 mo preceding
outcome assessment
Respiratory (noncancer)
- pulmonary function
Measures
Consideration of
confounding
Relevant exposure
time window(s)
• Standard protocol
• Age, sex, height, smoking
• Up to 6 months preceding pulmonary function measures
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Ne urob ehavioral
Measures
Consideration of
confounding
Relevant exposure
time window(s)
Thyroid
Measures
Consideration of
confounding
Relevant exposure
time window(s)
Obesity
Measures
Consideration of
confounding
Relevant exposure
timewindow(s)
Diabetes and insulin
resistance
Measures
Consideration of
confounding
Relevant exposure
timewindow(s)
• Standardized assessment tool, validation studies for specific study
population (e.g., age group, geographic location)
• Blinding of assessor to exposure
• Age, sex, socioeconomic status
• In utero; early childhood
• Assay used and evidence from validation studies, if available
• Sensitivity/detection limits, coefficient of variation; number of samples
below LOD
• Time of day and season when samples for thyroid hormone (and TSH)
collected
• Age, sex, smoking, iodine, radiation exposure (consider if these are related
to exposure)
• Varies by lifestage (i.e., infants, children, adults)
• Source of data (e.g., measured or self-reported weight and height)
• Age, sex, ethnicity, caloric intake, physical activity (consider if these are
related to exposure)
• Not established (likely to be more than one, including in utero)
• Source of data (e.g., biomarkers of insulin or glucose, medical records,
report)
self-
• Age, sex, ethnicity
• Not established (likely to be more than one, including in utero)
2 2.4. STUDY CHARACTERISTICS THAT WILL BE CONSIDERED IN THE
3 FUTURE EVALUATION AND SYNTHESIS OF THE CRITICAL
4 EXPERIMENTAL STUDIES FOR BBP
5 Beyond the initial methodological screening described above in Section 2.2.2,
6 methodological aspects of a study's design, conduct, or reporting will be considered again in the
7 overall evaluation and synthesis of the pertinent data that will be developed for each health effect.
8 Some general questions that will be considered in evaluating experimental animal studies are
9 presented in Table 2-9. These questions are, for the most part, broadly applicable to all
10 experimental studies.
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1
2
Table 2-9. Questions and relevant experimental information for the
evaluation of experimental animal studies
Methodological
feature
Test animal
Experimental setup
Exposure
Endpoint evaluation procedures
Outcomes and data and reporting
Question(s) considered
Based on the endpoint(s) in question, are concerns raised regarding the
suitability of the species, strain, or sex of the test animals on study?
Are the timing, frequency and duration of exposure, as well as animal age
and experimental group allocation procedures/ group size for each endpoint
evaluation, appropriate for the assessed endpoint(s)?
Are the exposure conditions and controls informative and reliable for the
endpoint(s) in question, and are they sufficiently specific to the compound ol
interest?
Do the procedures used to evaluate the endpoint(s) in question conform to
established protocols, or are they biologically sound? Are they sensitive for
examination of the outcome(s) of interest?
Were data reported for all pre-specified endpoint(s) and study groups, or
were any data excluded from presentation/ analyses?
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Note: "Outcome" refers to findings from an evaluation (e.g., steatosis), whereas "endpoint" refers to the
evaluation itself (e.g., liver histopathology).
Evaluation of some specific methodological features identified in Table 2-9, such as
exposure, is likely to be relatively independent of outcome. Other methodological features, in
particular those related to experimental setup and endpoint evaluation procedures, are generally
outcome specific (i.e., reproductive and developmental toxicity). In general, experimental animal
studies will be compared against traditional assay formats (e.g., those used in guideline studies),
with deviations from the protocol evaluated in light of how the deviations could alter interpretation
of the outcome in question. A full evaluation of all critical studies will be performed as part of the
critical review and synthesis of evidence of hazard identification for each of the health endpoints
identified in the evidence tables presented in Section 3.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
2 3.PRELIMINARY EVIDENCE TABLES AND
3 EXPOSURE-RESPONSE ARRAYS
4 3.1. DATA EXTRACTION FOR EPIDEMIOLOGICAL AND EXPERIMENTAL
5 STUDIES: PREPARATION OF PRELIMINARY EVIDENCE TABLES
6 The evidence tables present data from studies related to a specific outcome or endpoint of
7 toxicity. At a minimum, the evidence tables include the relevant information for comparing key
8 study characteristics such as study design, exposure metrics, and dose-response information.
9 Evidence tables will serve as an additional method for presenting and evaluating the suitability of
10 the data to inform hazard identification for butyl benzyl phthalate (BBP) during the analysis of
11 hazard potential and utility of the data for dose-response evaluation. For each critical study
12 selected, key information on the study design, including characteristics that inform study quality,
13 and study results pertinent to evaluating the health effects from subchronic and chronic oral
14 exposure to BBP are summarized in preliminary evidence tables.
15 Epidemiological studies are presented first where each study per table is listed in reverse
16 chronological order. Animal studies are then presented where each study per health endpoint is
17 presented in order by duration, followed by species and strain. Finally, animal metabolite studies
18 are also presented as monobutyl phthalate (MBP) and monobenzyl phthalate (MBzP) are thought to
19 contribute to developmental toxicity. Inclusion of these studies may help to inform the hazard
20 identification for BBP. Most results are presented as the percent change from the control group; an
21 asterisk (*) indicates a result that has been calculated and reported by study authors to be
22 statistically significant compared to controls (p <0.05). Unless otherwise noted in a footnote, doses
23 presented in the animal evidence tables were those reported by the study authors.
24 The information in the preliminary evidence tables for BBP is also displayed graphically in
25 preliminary exposure-response arrays. In these arrays, a significant effect (indicated by a filled
26 circle) is based on statistical significance by the study authors. The complete list of references
27 considered in preparation of these materials can be found on the Health and Environmental
28 Research On-line (HERO) website at http://hero.epa.gov/BBP and http://hero.epa.gov/phthalates-
29 humanstudies.
30
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
l 3.2. EPIDEMIOLOGICAL STUDIES
2 3.2,1. Sexual Differentiation Measures
3 Table 3-1. Evidence pertaining to BBP and sexual differentiation effects in
4 humans
Reference and study design
Results
Anogenital distance (AGD)
Suzuki et al. (2012) (Japan)
Population: 111 male infants from birth cohort
study, time period not given
Outcome: AGD measured 1-3 d after birth (AGD1 to
anterior genitalia, mean 45.8 mm, 14.8 mm/kg; AGD2
to posterior genitalia, mean 20.3 mm, 6.6 mm/kg)
Exposure: Maternal urine samples, mean 29 wks of
gestation
MBzP in urine (ng/mL):
Median 75th percentile
Unadjusted 3.57 8.73
SG-adjusted 4.73 10.8
Analysis: Linear regression considering gestational
week, birth order, maternal age, maternal smoking
during pregnancy, maternal environmental tobacco
smoke exposure, maternal urinary daidzein (soy
isoflavone) and equol (a urinary metabolite of
daidzein) concentrations, and environmental tobacco
smoke (smoking status of husbands of nonsmoking
women) as potential confounders
Association between MBzP and AGD measures reported
as not statistically significant (quantitative results not
reported)
Swan (2008) (United States; Minnesota, Missouri,
California)
Population: 106 boys from birth cohort study (SFF),
2000-2002, mean age 12.8 mo (0-36 mo)
Outcome: AGD (to posterior genitalia) measured at
0-36 mo (mean 70.4 mm, 7.1 mm/kg)
Exposure: Maternal urine sample, 3rd trimester
MBzP in urine (ng/mL):
Median 75th percentile
Unadjusted 8.3 23.5
Analysis: Regression analysis using mixed model
adjusting for age and weight percentile
Related references: Swan et al. (2005) (exposure
data and analysis of smaller sample size with less
robust method of adjustment for variation by size)
Percent change in AGD per interquartile increase in MBzP
concentration (p-value)
MBzP
-0.4 (0.826)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Cryptorchidism or testicular position
Swan (2008) (United States; Minnesota, Missouri,
California)
Population: 106 boys from birth cohort study (SFF),
2000-2002, mean age 12.8 mo (0-36 mo)
Outcome: Incomplete testicular descent assessed at
clinical exam (10% prevalence)
Exposure: Maternal urine sample, 3rd trimester
MBzP in urine (ng/mL):
Median 75th percentile
Unadjusted 8.3 23.5
Analysis: Logistic regression, adjusting for age and
weight percentile
Related references: Swan et al. (2005) (exposure
data)
MBzP reported as not associated with testicular position
(quantitative results not reported)
Main et al. (2006) (Denmark. Finland)
Population: 62 cases, 68 controls from two
pregnancy cohorts, born 1997-2001, age 3 mo
Outcome: Cryptorchidism, at birth and/or 3 mo
Exposure: Breast milk samples collected 1-3 mo of
age
MBzP in breast milk (u.g/L), all samples:
Median (range)
Denmark 0.9 (0.2-14)
Finland 1.3 (0.4-26)
Analysis: Mann-Whitney U-test for comparison of
MBzP concentrations in boys with and without
Cryptorchidism
Median MBzP in breast milk (u.g/L)
Controls
1.20
(p >0.40)
Cases
1.25
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Infant hormone levels
Lin etal. (2011a) (Taiwan)
Population: 155 infants (81 boys, 74 girls) from birth
cohort, born 2000-2001
Outcome: Cord blood hormone levels
Exposure: Maternal urine sample 3rd trimester
MBzP in urine (percentile):
Median 75th 95th
Unadjusted (ng/mL) 8.85 15.1 40.3
Cr-adjusted (u.g/g Cr) 15.6 25.9 43.9
Analysis: Pearson correlation analysis and linear
regression adjusted for variables shown in the results
column
Pearson correlation coefficient (r) and regression
coefficient (P), log-MBzP (ng/g Cr) and cord blood
hormone level (regression adjusted for maternal age, BMI,
smoking habit, gestational age, parity, and use of
contraceptive drugs)
Boys
Free testosterone (ng/dL)
Estradiol (pg/mL)
Free testostone:estradiol ratio
Girls
Free testosterone (ng/dL)
Estradiol (pg/mL)
Free testostone:estradiol ratio
NR = not reported
*p<0.10; all other p-values>0.10
r p
0.05 NR
0.14 0.11
-0.03 -0.01
-0.18 NR
-0.20* 0.00
-0.10 0.10
Main et al. (2006) (Denmark. Finland)
Population: 130 male infants from two pregnancy
cohorts (cryptorchidism cases and controls combined
for this analysis), born 1997-2001, age 3 mo
Outcome: Serum steroidal and gonadotropin
hormone levels in infants, samples collected when
breast milk samples delivered to hospital
Exposure: Breast milk samples collected 1-3 mo of
age
MBzP in breast milk (ng/L), all samples:
Median (range)
Denmark 0.9 (0.2-14)
Finland 1.3 (0.4-26)
Analysis: Cases and controls combined for analysis
of association between metabolite concentration and
hormone analysis using partial Spearman correlation
coefficients adjusted for country of birth
Spearman correlation coefficient (p-value), MBzP (ng/L)
and serum hormone level (n = 96 boys)
SHBG (nmol/L)
Free testosterone (nmol/L)
Testosterone (nmol/L)
LH (IU/L)
FSH (IU/L)
0.188 (0.074)
-0.007 (0.951)
0.115 (0.271)
0.049 (0.643)
0.045 (0.668)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Gender-related play
Swan et al. (2010) (United States; Minnesota,
Missouri, California, Iowa)
Population: 145 children from birth cohort study
(SFF), 2000-2002 and 2002-2005 (Iowa), ages
4-7 yrs; second follow-up
Outcome: Gender-specific play based on Pre-School
Activities Inventory (24 items completed by parent or
caregiver; subscores of male-oriented items and
female-oriented items and a composite score
consisting of male summation minus the female
summation scores)
Exposure: Maternal urine sample, 3rd trimester
MBzP in urine (ng/mL); distribution not reported for
this analysis; EPA assumed similar distribution as
seen in Swan et al. (2005)
MBzP in urine (ng/mL):
Median 75th percentile
Unadjusted 8.3 23.5
Analysis: Regression analysis using Generalized
Linear Models, considering creatinine, sex and age of
child, maternal age, parental education, number of
same and opposite sex siblings, ethnicity, clinic
location, and parental attitude as potential covariates
Related references: Swan et al. (2005) (exposure
data)
log-MBzP reported as not associated with masculine or
composite activity score (quantitative results not
reported)
1
2
3
BMI = body mass index; FSH = follicle stimulating hormone; LH= luteinizing hormone; SFF = Study for Future
Families; SHBG = sex-hormone binding globulin
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.2.2. Male Reproductive Effects in Humans
2
3
Table 3-2. Evidence pertaining to BBP and reproductive hormones in adult
men
Reference and study design
Results
Jurewicz et al. (2013) (Poland)
Population: 269 men from infertility clinic with normal
sperm concentration (20-300 million/mL) or slight
oligozoospermia (15-20 million/mL); mean age 32 yrs;
time period not reported; MBzP measured in 268 samples
Outcome: Plasma testosterone, E2, and FSH
Exposure: Urine sample, collected at same time as
plasma sample
MBzP in urine:
Geometric mean (SD)
Unadjusted (u.g/L) 8.3 (3.2)
Cr-adjusted (u.g/g Cr) 6.9 (3.5)
Analysis: Linear regression, adjusting for variables shown
in results column
Regression coefficient (p-value) for increase in
hormone unit change in log = MBzP (adjusted for age,
smoking, medical history [mumps, cryptorchidism,
testes surgery, testes trauma], abstinence time, and
urinary creatinine)
Testosterone (ng/mL)
E2 (pg/mL)
FSH (IU/L)
-0.09 (0.52)
0.87(0.11)
0.26 (0.096)
Joensen et al. (2012) (Denmark)
Population: 881 men from general population, assessed
at military conscript exam*, 2007-2009, median age
19.1 yrs (5th-95th percentile: 18.4-22.0 yrs)
Outcome: Serum steroidal and gonadotropin hormones
Exposure: Urine sample, collected at same time as serum
sample for hormone analysis
MBzP in urine (ng/mL):
Median 95th percentile
Unadjusted 34 164
Analysis: Linear regression considering age, BMI,
smoking, alcohol consumption, time of blood sampling,
assay type, ethnicity, BMI squared, in utero exposure to
tobacco smoke, previous or current diseases, recent
fever, and recent use of medication as potential
covariates
*As reported by Ravnborg et al. (2011)
Results for individual phthalate metabolites
(including MBzP) reported as "few significant
associations" with free testosterone, estradiol, SHBG,
LH, inhibin-B, or FSH (quantitative results not
reported); analyses adjusted for age, BMI, smoking,
alcohol consumption, and time of blood sampling
(and assay type for inhibin-B only)
Mendiola et al. (2012) (United States; Minnesota,
Missouri, California, Iowa, Boston)
Populations: 425 fertile men with pregnant partners
enrolled in birth cohort study (SFF), 1999-2005; mean age
32 yrs; 425 men who were male partners of infertile
couples seeking evaluation (MGH); 2000-2004, mean age
36 yrs
Outcome: Serum steroidal and gonadotropin hormones
Exposure: Urine sample, collected at same time as serum
sample for hormone analysis
MBzP in urine (ng/mL):
Median 90th percentile
SFF: Unadjusted 12.5 49.8
Authors report "no associations between any
hormone levels [testosterone, estradiol, SHBG, LH,
inhibin-B, or FSH] and any urinary metabolites of
phthalates other than DEHP" [including MzBP]
(quantitative results not reported)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
MGH: Unadjusted 8.2 24.9
All: Unadjusted 9.8 41.2
Analysis: Pearson correlation coefficients of
log(10)-transformed MzBP and hormone measures
(bivariate analysis); linear regression considering age, age
square, BMI, smoking status, ethnicity, urinary creatinine
concentration (SFF models) or specific gravity (MGH
models), time of sample collection, time of collection
squared, and study center (SFF vs MGH) for each
population separately and for the pooled population
Related references: This is a pooled analysis of a study of
fertile men (Mendiola et al., 2011) and men from infertile
couples (Meeker etal., 2009a)
Mendiola et al. (2011) (United States; Minnesota,
Missouri, California, Iowa, New York)
Population: 425 fertile men with pregnant partners
enrolled in birth cohort study (SFF), 1999-2005, mean age
32yrs
Outcome: Serum steroidal and gonadotropin hormones
Exposure: Urine sample, collected at same time as serum
sample for hormone analysis; data reported in Mendiola
etal. (2012)
MBzP in urine (ng/mL):
Median 90th percentile
Unadjusted 12.5 49.8
Analysis: Pearson correlation coefficients of
log(10)-transformed MzBP and hormone measures
(univariate analysis); linear regression considering age,
age square, BMI, smoking status, ethnicity, urinary
creatinine concentration, time of sample collection, and
time of collection squared
Authors report "little or no association with
metabolites of phthalate other than DEHP" [including
MzBP] with testosterone, estradiol, SHBG, LH,
inhibin-B, or FSH (quantitative results not reported)
Meeker et al. (2009a) (United States; Boston)
Population: 425 men from subfertility clinic, 2000-2004;
mean age36yrs
Outcome: Serum steroidal and gonadotropin hormones
Exposure: Urine sample, collected at same time as serum
sample
MBzP in urine (ng/mL) (percentile):
Median 75th percentile 95th percentile
SG-adjusted 8.20 15.9 40.6
Analysis: Linear regression using untransformed
(testosterone, estradiol) or natural logarithm transformed
(free androgen index, FSH, LH) hormone levels;
considering age, BMI, smoking status, race, previous
infertility example, prior ability to impregnate partner,
and season and time of sample collection as potential
covariates
Related references: Duty et al. (2005)
Regression coefficient (95% Cl) for change in
hormone with IQR increase in adjusted MBzP
concentration (adjusted for age, BMI, smoking,
season and time of day sample was collected, and
[for testosterone and estradiol only] SHBG)
Untransformed hormone level (0.0 = no effect)
Testosterone (ng/dL) 4.58 (-7.91,17.0)
Estradiol (pg/mL) -0.21 (-1.53,1.09)
Inhibin B (pg/mL) 1.81 (-6.54,10.2)
Ln-transformed hormone level (1.0 = no effect)
Free androgen index 1.03 (0.99,1.07)
FSH (IU/L) 0.98 (0.92, 1.04)
LH (IU/L) 1.00 (0.95, 1.05)
SHBG (nmol/mL) 1.00 (0.95, 1.04)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Prolactin (ng/mL)
1.01 (0.96,1.06)
Jonsson et al. (2005) (Sweden)
Population: 234 men from general population, assessed
at military conscription exam in 2000; ages 18-21 yrs
Outcome: Serum steroidal and gonadotropin hormones
Exposure: Urine sample, collected at same time as serum
sample for hormone analysis
MBzP in urine (percentile):
Median 75th 95th
Unadjusted (ng/mL) 16 37 74
Adjusted (nmol/mmol Cr) 4.4 7.6 19
Analysis: Mean difference between high and low
quartiles
Mean difference (95% Cl), highest (>7.71 nmol/mmol
Cr) compared with lowest quartile of MBzP
(<1.10 nmol/mmol Cr)
Testosterone (nM)
Free testosterone (T/SHBG)
Estradiol (pM)
FSH (IU/L)
LH (IU/L)
-0.03 (-2.1,2.0)
0.06 (-0.05, 0.2)
0.7 (-5.3, 6.7)
0.1 (-0.5, 0.7)
0.4 (-0.2, 1.0)
1
2
3
Cl = confidence interval; DEHP = di(2-ethylhexyl)phthalate; E2 = estradiol; IQR = interquartile range; MGH =
Massachusetts General Hospital; SD = standard deviation
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.2.3. Male Pubertal Development in Humans
2
3
Table 3-3. Evidence pertaining to BBP and the timing of male puberty or sex
hormones in boys
Reference and study design
Ferguson et al. (2014b) (Mexico)
Population: 115 boys ages 8-14 yrs from a birth
cohort (Early Life Exposure in Mexico to
Environmental Toxicants, participants enrolled
during first trimester 1994-2004); follow up
initiated in 2010
Outcome: Adrenarche or puberty, based on
Tanner staging by physician (pubic hair stage >2;
genitalia stage >2 or testicular volume >3 mL);
serum hormone level
Exposure: Maternal urine sample (n = 107) from
third trimester or child's urine sample (n = 113)
collected at time of Tanner staging and serum
collection
Unadjusted MBzP in urine (ng/mL):
Median 95th percentile
Maternal sample 5.20 15.4
Child's sample 5.60 19.9
Analysis: Logistic regression for analysis of
puberty onset, adjusting for variables shown in
results column; linear regression for analysis of
hormone levels, considering age, BMI z-score,
socioeconomic status, and maternal smoking
potential covariates
Results
OR (95% Cl) for adrenarche or puberty per interquartile
increase in In-transformed MBzP (adjusted for child age, BMI
z-score, and urine specific gravity)
Exposure basis
Tanner stage
or testicular Maternal urine
volume (prenatal) Child urine
Pubic hair 0.27(0.08,0.94) 0.73(0.21,2.58)
(stage >2)
Genitalia 0.76(0.47,1.23) 1.71(0.78,3.76)
(stage >2)
Testicular 0.76(0.41,1.41) 2.17(0.80,5.87)
volume
(>3 mL)
Percent change (95% Cl) in serum hormone level per
interquartile increase in In-transformed MzBP (adjusted for
urine specific gravity, child age, and BMI z-score)
Exposure basis
Serum Maternal urine
hormone (prenatal) Child urine
Testosterone 3.82 (-18.4, 32.1) -23.5 (-47.3, 11.1)
Free -3.21 (-24.6, 24.3) -28.3 (-51.5, 6.04)
testosterone
SHBG 11.0 (2.33, 20.3) 7.77 (-5.56, 23.0)
DHEAS -3.35 (-14.0, 8.58) 8.49 (-9.56, 30.2)
Estradiol -1.18 (-8.36, 6.57) -10.2(-20.1, 0.96)
Inhibin B -4.81 (-12.8, 3.95) 9.50 (-4.40, 25.4)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Mouritsen etal. (2013b) (Denmark)
Population: 53 boys from population-based cohort
(COPENHAGEN Puberty Study), 2006-2010; age
llyrs
Outcome: Adrenarche or puberty, based on
Tanner staging by physician (pubarche = pubic hair
stage >2 and testicular volume >3 mL); serum
hormone level
Exposure: Urine sample, first morning sample;
data reported in Mouritsen et al. (2013a),
Supplemental Material
MBzP in urine (ng/mL):
Geometric mean Maximum
49 1,660
(based on larger sample of 84 boys)
Analysis: Two-tailed Mann-Whitney U-test for
comparisons between groups, comparing median
hormone levels and pubertal stage in "high" and
"low" phthalate groups (based on above or below
group mean excretion)
Median age (yrs) at development by MBzP level
Pubarche
(pubic hair
stage >2)
Testicular
volume >3 mL
Low
12.1
11.6
High
11.4
11
Median hormone concentration by MBzP level
Testosterone
(nmol/L)
DHEAS
(umol/L)
Adione
(nmol/L)
Estradiol
(pmol/L)
FSH (IU/L)
LH (IU/L)
Low
<0.23
2.14
1.46
High
<0.23
1.33
(p<0.05)
1.13
1.38
0.25
1.5
0.28
Mieritz et al. (2012) (Denmark)
Population: 38 boys with pubertal gynecomastia
and 190 age-matched controls drawn from
555 boys from population-based cohort
(COPENHAGEN Puberty Study), 2006-2008; ages
6-19 yrs
Outcome: Anthropometry, pubertal stage (pubic
hair and genital development), presence of
gynecomastia, and serum testosterone
Exposure: Urine sample, first morning sample
MBzP in urine (ng/mL):
Median 95th percentile
Groups 47.70 219.2
(boys without gynecomastia, all ages)
Analysis: Two-tailed Mann-Whitney U-test for
comparisons between groups; linear regression
with age adjustment for association with serum
testosterone; probit analysis with phthalate
concentrations divided in quartiles for analysis of
puberty timing
MBzP concentration (ng/mL) by group
Median
95th percentile
Group 1
(n = 38)
56.79
211.0
Group 2
(n = 189)
47.20
185.3
Group 3
(n = 517)
47.70
219.2
Group 1 = boys with palpable gynecomastia
Group 2 = boys without palpable gynecomastia (age-
matched)
Group 3 = boys without palpable gynecomastia (all ages)
No association between MBzP concentration and timing of
puberty or serum testosterone level (quantitative results not
reported)
1
2
3
OR= odds ratio
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.2.4. Semen Parameters and Infertility
2
3
Table 3-4. Evidence pertaining to BBP and semen parameters or infertility in
adult men or couples
Reference3 and Study Design
Results
Kranvogl et al. (2014) (Slovenia)
Population: 136 men from couples seeking
infertility treatment (mean age 36.2 yrs, range
24-54 yrs), 2012
Outcome: Semen analysis
Exposure: Urine sample, collected at same time
as semen sample
MBzP in urine
Unadjusted (u.g/L) 3.5 20.7
Cr-adjusted (u.g/g Cr) 2.9 15.5
Analysis: Spearman correlation
Spearman correlation coefficient, MBzP and sperm
parameters:
Sperm concentration
Sperm motility
(p >0.05 for both parameters)
-0.014
0.058
(Jurewicz et al. (2013)) (Poland)
Population: 269 men from infertility clinic with
normal sperm concentration (20-300 million/mL)
or slight oligozoospermia (15-20 million/mL);
mean age 32 yrs; time period not reported; MBzP
measured in 268 samples
Outcome: Semen analysis
Exposure: Urine sample, collected at same time
as semen sample
MBzP in urine:
Geometric mean (SD)
Unadjusted (u.g/L) 8.3 (3.2)
Cr-adjusted (u.g/g Cr) 6.9 (3.5)
Analysis: Linear regression, adjusting for
variables shown in results column
Regression coefficient (p-value) for change in sperm
parameter with unit change in log-MBzP (adjusted for age,
smoking, medical history [mumps, cryptorchidism, testes
surgery, testes trauma], abstinence time, and urinary
creatinine)
Log-transformed sperm concentration -0.07 (0.25)
(million/mL)
Sperm motility (%) 1.86 (0.10)
Abnormal sperm morphology (%) 1.17 (0.28)
DNA fragmentation index -0.05 (0.20)
Joensen et al. (2012) (Denmark)
Population: 881 men from general population,
assessed at military conscript exam*, 2007-2009,
median age 19.1 yrs (5th-95th percentile:
18.4-22.0 yrs)
Outcome: Semen analysis
Exposure: Urine sample, collected at same time
as semen sample
MBzP in urine (ng/mL):
Median 95th percentile
Unadjusted 34 164
Analysis: Linear regression, considering age,
BMI, smoking, alcohol consumption, ethnicity,
BMI squared, in utero exposure to tobacco
smoke, previous or current diseases, recent
fever, recent use of medication, abstinence time,
and time from ejaculation to analysis as potential
covariates
Results for individual phthalate metabolites (including MBzP)
reported as "few significant associations" with sperm volume,
count, or percentage progressively motile sperm (quantitative
results not reported; analyses adjusted for abstinence time
[volume, concentration, and count] or time from ejaculation
to analysis [progressively motile]; percent of morphologically
normal sperm was left unadjusted)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference3 and Study Design
Results
"As reported by Ravnborg et al. (2011)
Liu etal. (2012) (China)
Population: 97 men from subfertility clinic,
2009-2010; mean age 32 yrs
Outcome: Semen analysis; results dichotomized
above and below WHO reference values; n = 43
with normal semen parameters
Exposure: Urine sample, collected at same time
as semen sample
MBzP in urine:
Median 66th percentile
Unadjusted (ng/mL) 3 servings/wk), and
specific gravity
°Adjusted for specific gravity
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference3 and Study Design
Results
Analysis: Dichotomized outcomes (above and
below WHO reference values), MBzP
dichotomized at median or divided into tertiles;
age, education (3 levels), income (3 levels), race,
BMI (3 levels), current smoking status, and
alcohol use (2 levels) considered as potential
confounders; specific gravity also included in all
models
Results of tertile analysis not reported
Mauser et al. (2007) (United States; Boston)
Population: 379 male partners from subfertility
clinic, 2000-2004; mean age 36 yrs
Outcome: Sperm DNA damage assessed by
neutral comet assay
Exposure: Urine sample, collected at same time
as semen sample
MBzP in urine (ng/mL) (percentile):
Median 75th 95th
SG-adjusted 7.9 15.0 46.2
Analysis: Linear regression, considering age,
abstinence time, smoking status, and race as
potential covariates
Related reference: Duty et al. (2003b)
Regression coefficient (95% Cl) for DNA damage associated
with interquartile range increase in In-MBzP (adjusted for age
and smoking status)
Comet extent
(urn)
Tail distribution
(urn)
5.12(0.98,9.25) 2.49(0.82,4.13)
%DNAtail
0.11 (-1.56, 1.77)
Mauser et al. (2006) (United States; Boston)
Population: 443 male partners from subfertility
clinic 2000-2004; mean age 36 yrs
Outcome: Semen analysis; results dichotomized
above and below WHO reference values
Exposure: Urine sample, collected at same time
as serum sample for hormone analysis
MBzP in urine (ng/mL) (percentile):
Median 75th 95th
SG-adjusted 8.0 15.5 40.6
Analysis: Logistic regression, considering age,
race, BMI, abstinence time, and smoking as
potential covariates
Related references: (Mauser etal. (2005): Duty
et al. (2004): Duty et al. (2003a))
OR (95% Cl) by quartile of MBzP (ng/mL) (adjusted for age,
abstinence time, and smoking; comparison group = 210 men
without deficiencies on any of these three parameters)
MBzP
quartile
1 (low)
2
3
4 (high)
(trend p)
Sperm Sperm
concentration motility
<20xl06/mL <50% motile
1.0 (referent) 1.0 (referent)
1.1 (0.4, 2.6) 1.3 (0.7, 2.3)
1.1 (0.4, 2.5)
1.9 (0.8, 4.3)
(0.13)
1.3 (0.8, 2.3)
1.3 (0.7, 2.3)
(0.36)
Sperm morphology
<4% normal
1.0 (referent)
0.7 (0.3, 1.4)
0.9 (0.4, 1.7)
1.1(0.6,2.1)
(0.76)
Regression coefficient (95% Cl) for sperm motion parameters
by quartile of MBzP (ng/mL) (adjusted for age, smoking, and
abstinence time)
MBzP
(ng/mL)
quartile
1 (low)
2
Straight line
velocity
(u.m/s)
Curvilinear
velocity
(u.m/s)
1.0 (referent) 1.0 (referent)
0.66
(-2.01, 3.34)
0.11
(-2.59, 2.81)
1.44
(-3.10,5.99)
1.29
(-3.29, 5.88)
Linearity (%)
1.0 (referent)
-0.23
(-2.12, 1.66)
-1.13
(-3.04, 0.77)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference3 and Study Design
Results
4 (high)
(trend p)
-1.31
(-3.98,1.36)
(0.29)
-1.20
(-5.73, 3.34)
(0.60)
-0.69
(-2.58, 1.20)
(0.33)
MBzP quartile cut points: 0.04-4.2, 4.2-8.0, 8.0-15.3,
15.5-540.2 ng/mL
No interaction with PCBs
Jonsson et al. (2005) (Sweden)
Population: 234 men from general population,
assessed at military conscription exam in 2000;
ages 18-21 yrs
Outcome: Semen analysis
Exposure: Urine sample, collected at same time
as semen sample
MBzP in urine (percentile):
Median 75th 95th
Unadjusted (ng/mL) 16 37 74
Adjusted (nmol/mmol Cr) 4.4 7.6 19
Analysis: Mean difference between high and low
quartiles
Mean difference (95% Cl), highest (>7.71 nmol/mmol Cr)
compared with lowest (<1.10 nmol/mmol Cr) quartile MBzP
(positive difference indicates lower value in highest exposure
quartile)
Sperm concentration (x 106/mL) 7.2 (-16, 31)
Sperm motility (%) -4.3 (-10,1.6)
Sperm damage (chromatin integrity) -0.3 (-3.7, 3.1)
Infertility
Buck Louis et al. (2014) (United States; Michigan
and Texas)
Population: 501 couples discontinuing
contraception and attempting to achieve
pregnancy; recruited from 16 counties using
population sampling. Women's mean age
30.0 yrs, men's mean age 31.8 yrs; 2005-2009
Outcome: Time to pregnancy as assessed by
diaries recording intercourse and menstruation,
home-fertility monitoring to detect ovulation,
and home pregnancy tests
Exposure: Urine samples from both partners,
collected at enrollment (beginning of pregnancy
attempt)
Unadjusted MBzP in urine (ng/mL) among
couples achieving pregnancy:
Geometric mean (95% Cl)
Women 4.61(4.06-5.23)
Men 2.79(2.44-3.19)
Analysis: Fecundability ORs calculated using Cox
models, adjusting for variables shown in results
column
Fecundability OR (95% Cl) per unit increase in log-transformed
MBzP scaled by SD (adjusted for female age, difference in
couples' ages, research site, and both partners' urinary
creatinine, BMI, and serum cotinine; in addition, results for
exposure in each partner adjusted for exposure in the other
partner, and models accounted for left truncation or time off
contraception)
Women
Men
0.98 (0.81,1.20)
0.80 (0.67, 0.97)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference3 and Study Design
Results
Tranfo et al. (2012) (Italy)
Population: 56 infertile couples from assisted
reproduction center, 56 fertile couples (parents
of one or more children, living in same area),
time period not reported; mean age 39-40 yrs in
both groups
Outcome: Primary or secondary infertility as
assessed by WHO criteria (cause attributed to
males in 8/56 couples)
Exposure: Urine sample
MBzP in urine, fertile couples:
Median 95th percentile
Cr-adjusted (jjg/g Cr) 8.8 85.32
Analysis: Mann-Whitney U-test for comparison
of MBzP concentrations by group
MBzP concentration in urine (ng/g Cr) in fertile and infertile
couples
Median
95th percentile
Fertile
8.80
85.32
Infertile p-value
12.37 0.009
88.10
Sex-stratified comparison was similar for men and for women,
though the p-value was slightly higher than 0.05 (quantitative
results not reported)
1
2
3
DNA = deoxyribonucleic acid; LOD = level of detection; PCB = polychlorinated biphenyl; WHO = World Health
Organization
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.2,5. Female Reproductive Effects in Humans
2
3
Table 3-5. Evidence pertaining to BBP and reproductive hormones in adult
women
Reference and study design
Results
Maternal hormones during pregnancy
Sathvanaravana et al. (2014) (United States; Minnesota,
Missouri, California)
Population: 180 mothers from birth cohort (SFF), recruited
during pregnancy, 1999-2002
Outcome: Serum hormone levels, samples collected
during prenatal clinic visit
Exposure: Maternal urine sample, collected during 2nd or
3rd trimester
MBzP in urine (ng/mL):
Median 75th percentile
Unadjusted 11.0 38.6
Analysis: Linear regression, log-transformed MBzP, and
log-transformed hormone level
Regression coefficient (95% Cl) for change in
maternal log-transformed serum hormone level
with unit increase in log-transformed MBzP,
stratified by sex of fetus
Testosterone
(total)
Testosterone
(free)
Estradiol
Mothers with
male fetus
(n = 94)
0.06
(-0.07,0.19)
0.07
(-0.07, 0.21)
-0.03
(-0.12,0.07)
Mothers with
female fetus
(n = 86)
-0.13
(-0.26, 0.01)
-0.10
(-0.25, 0.04)
-0.10
(-0.23, 0.03)
Hart et al. (2013) (Australia)
Population: 123 mothers from birth cohort (Western
Australian Pregnancy Cohort), whose mothers were
recruited at 18 wks of gestation between 1989 and 1991
Outcome: Reproductive and gonadotropin hormone levels
in maternal serum collected at 18 and 34-36 wks of
gestation
Exposure: Maternal serum samples (n = 123) collected at
18 and 34-36 wks of gestation (combined aliquot from
both time periods)
MBzP in serum (ng/mL):
Median 90th percentile
MBzP 1.26 3.87
Analysis: Correlation between quartiles of serum MBzP
and log-transformed hormone levels
Correlation coefficient between log-transformed
maternal serum hormone level and quartiles of
MBzP in maternal serum
Androstene-
dione (nmol/L)
DHEAS (u.mol/L)
Testosterone
(pmol/L)
SHBG (nmol/L)
Free
testosterone
(pmol/L)
Free
testosterone
index
At 18 wks of
gestation
(n = 119)
-0.006
-0.057
-0.009
-0.123
0.037
0.053
At 34-36 wks of
gestation
(n = 114)
-0.045
-0.132
-0.063
-0.149
0.027
0.033
p >0.10 for all correlations
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.2,6. Female Pubertal Development in Humans
2
3
Table 3-6. Evidence pertaining to BBP and timing of female puberty or sex
hormones in girls
Reference and study design
Results
Precocious puberty or thelarche
Chenetal. (2013) (Taiwan)
Population: 71 girls with central precocious puberty
from pediatric endocrinology clinic and 29 controls
from schools recruited 2006-2009; mean ages 8.1 and
6.8yrs, respectively
Outcome: Premature puberty based on appearance
of thelarche, pubic hair or menarche before 8 yrs of
age; Tanner staging and serum levels of LH releasing
hormone used for additional classification
Exposure: Urine sample (child's), collected at same
time as clinical assessment
MBzP in urine of controls:
Mean (95% Cl)
Unadjusted (ng/mL) 2.45 (0.70,18.4)
Cr-adjusted (u.g/g Cr) 3.74 (0.95, 50.4)
Analysis: MBzP concentrations in cases and controls
compared with Mann-Whitney U-test
Mean (95% Cl) MBzP in cases and controls
Controls Cases
Unadjusted
(ng/mL)
Cr-adjusted
Cr)
2.45
(0.70, 18.4)
3.74
(0.95, 50.4)
6.22
(0.70, 167)
9.0
(1.14, 172)
(p-value)
(0.002)
(0.005)
Frederiksen et al. (2012) (Denmark)
Population: 24 girls with precocious puberty (n = 13
with central precocious puberty, n = 6 with early
normal puberty, n = 5 with premature thelarche) from
outpatient clinic, 2008-2009 and 184* age-matched
controls from population-based cohort (COPENHAGEN
Puberty Study), recruited from high schools
2006-2008; age 7.4-9.9 years
Outcome: Precocious puberty, early normal puberty,
or premature thelarche, based on Tanner staging by
physician
Exposure: Urine sample (child's), collected at clinical
evaluation
MBzP in urine (ng/mL), controls:
Median 95th percentile
Unadjusted 48 212
(based on larger sample of 725 controls)
Analysis: MBzP concentrations in cases and controls
compared with Mann-Whitney U-test
*Study reports number of controls inconsistently; text
reports 164 controls, while Table 4 reports 184
Median (range) MBzP (ng/mL) in cases and controls
Controls
54 (4.6-779)
Precocious
puberty
38(7.5-177)
(p-value)
(<0.05)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Lomenicketal. (2010) (United States, Ohio and
Kentucky)
Population: 28 girls with central precocious puberty,
28 age- and race-matched controls; all recruited from
pediatric endocrinology clinic, 2005-2008; mean age
7 yrs
Outcome: Central precocious puberty defined based
on clinical standards (appearance of physical
characteristics of puberty before 8 yrs of age, with
laboratory confirmation of central origin of breast
development); no cases had received medical
treatment prior to urine sample collection
Exposure: Urine sample (child's), collected at clinical
evaluation
MBzP in urine of controls:
MeaniSE
Unadjusted (ng/mL) 42.8 ± 8.8
Cr-adjusted (u.g/g Cr) 40.5 ± 7.1
Analysis: MBzP concentrations in cases and controls
compared with Wilcoxon rank-sum test
Mean ± SE MBzP in cases and controls
Central
precocious
Controls puberty (p-value)
Unadjusted 42.8 ± 8.8 40.2 ± 8.4 (0.81)
(ng/mL)
Cr-adjusted 40.5 ±7.1 50.6 ± 11.4 (0.92)
Cr)
Chou et al. (2009) (Taiwan)
Population: 30 girls with premature thelarche and
26 girls with central precocious puberty from pediatric
endocrinology clinic; 33 controls from school exams;
mean ages 6.7, 8.0, and 8.2 yrs, respectively, in the
groups, time period not reported
Outcome: Premature puberty based on appearance
of any physical characteristics of puberty before 8 yrs
of age
Exposure: Urine sample (child's) collected at same
time as clinical assessment
MBzP in urine (ng/mL), controls:
Mean ±SD
Unadjusted 3.3 ±6.1
Analysis: One-way ANOVA comparing MBzP
concentrations between groups
Unadjusted MBzP in urine; mean ± SD (ng/mL)
Central precocious Premature thelarche
puberty cases cases
Controls
3.3 ±6.1 12.7 ±33.0
p >0.3 for comparison with controls
7.4 ±9.4
Pubertal development (general population)
Hart et al. (2013) (Australia)
Population: 121 girls from birth cohort study
(Western Australian Pregnancy Cohort), whose
mothers were recruited at 18 wks of gestation
1989-1991; follow-up at ages 14-16 yrs
Outcome: Age at menarche
Exposure: Maternal serum samples (n = 123)
collected at 18 and 34-36 wks of gestation (combined
aliquot from both time periods)
MBzP in serum (ng/mL):
Median 90th percentile
Unadjusted 1.26 3.87
Authors reported no association between MBzP and age
at menarche (quantitative results not reported)
Authors reported no correlation between MBzP and
serum SHBG, FSH, total testosterone, free androgen
index, anti-Mullerian hormone, or inhibin B in
adolescents (quantitative results not reported by study
authors)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Analysis: Correlation between log-transformed MBzP
and age at menarche
Mouritsen etal. (2013b) (Denmark)
Population: 47 girls from population-based cohort
(COPENHAGEN Puberty Study), 2006-2010; age 10 yrs
Outcome: Adrenarche or puberty, based on Tanner
staging by physician (pubarche = breast stage >2 and
pubic hair stage >2); serum hormone level
Exposure: Urine sample, first morning sample; data
reported in Mouritsen et al. (2013a), Supplemental
Material
Geometric mean Maximum
MBzP in urine (ng/mL): 37 433
(based on larger sample of 84 girls)
Analysis: Two-tailed Mann-Whitney U-test for
comparisons between groups, comparing median
hormone levels and pubertal stage in "high" and "low"
phthalate groups (based on above or below group
mean excretion)
Median age (yrs) at development by MBzP level
Low High
10.8 10.8
Pubarche (pubic
hair stage >2)
Pubarche (breast
stage >2)
10.5
10.2
Median hormone concentration by MBzP level
Testosterone
(nmol/L)
DHEAS (u.mol/L)
Adione (nmol/L)
Estradiol (pmol/L)
FSH (IU/L)
LH (IU/L)
Low
<0.23
1.03
1.63
19
2.12
0.08
High
<0.23
0.83
1.3
20
1.82
0.11
Frederiksen et al. (2012) (Denmark)
Population: 725 healthy girls ages 5.6-19.1 yrs from
COPENHAGEN Puberty Study cohort, recruited from
high schools during 2006-2008
Outcome: Stage of breast or pubic hair development;
Serum steroid and gonadotropin hormones
Exposure: Urine sample (child's), collected at time of
pubertal stage assessment
MBzP in urine (ng/mL), all 725 participants:
Median 95th percentile
Unadjusted 48 212
Analysis: Probit analysis, results verified using Pool-
Adjacent-Violators algorithm
Mean age (95% Cl) (yrs) at entry into breast stage 2 or
pubic hair stage 2, by quartile of MBzP
MBzP Breast stage 2
quartile (n = 394)
l(low) 9.66(9.16,10.14)
2 9.92 (9.44, 10.40)
3 10.10(9.63,10.55)
4 (high) 10.06 (9.59, 10.54)
Pubic hair stage 2 (n
not reported)
10.96 (10.67, 11.27)
11.25 (10.93, 11.58)
10.95 (10.68, 11.24)
11.39 (11.08, 11.72)
Levels of FSH, LH, and estradiol were similar across MBzP
exposure groups (quantitative results not reported). A
lower prevalence of detectable testosterone was seen
with increasing MBzP quartile; however, the association
did not remain significant after a correction was applied
for skewed age distribution between quartiles
(quantitative results not reported)
1
2
ANOVA= analysis of variance; DHEAS = dehydroepiandrosterone; SE = standard error
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.2,7. Gynecological Conditions in Humans
2
3
Table 3-7. Evidence pertaining to BBP and gynecological conditions in
humans
Reference and study design
Results
Endometriosis
Buck Louis et al. (2013) (United States, California and Utah)
Population: 473 women undergoing laparoscopy or
laparotomy and 127 population age- and residence-matched
referents, 2007-2009; ages 18-44 yrs; confirmed cases of
endometriosis matched to women without endometriosis
within each cohort: operative cohort 190 cases,
238 controls; population cohort 14 cases, 127 controls
Outcome: Endometriosis confirmed by surgery (operative
cohort) or MRI (population cohort)
Exposure: Urine sample
MBzP in urine (ng/mL), unadjusted:
Geometric mean
Operative cohort-controls 7.82
Population cohort-controls 6.46
Analysis: Student's t-test or Wilcoxon test for continuous
data; logistic regression, adjusting for age, BMI, and
creatinine; sensitivity analyses conducted restricting cohort
to endometriosis stages 3 and 4 diagnoses or visually and
histologically confirmed endometriosis, and referent group
consisting of women with postoperative diagnosis of normal
pelvis
OR (95% Cl) for endometriosis per unit increase in
In-MBzP, by cohort (adjusted for age, BMI, and
creatinine)
Operative cohort
Population cohort
0.84 (0.65,1.07)
1.47 (0.76, 2.85)
Adjusted OR (95% Cl) for endometriosis per unit
increase in In-MBzP in operative cohort (sensitivity
analysis)
Endometriosis stage 3 and 4
(n = 339)
Visual/histological confirmed
endometriosis (n = 473)
Comparison with women with
postoperative diagnosis
normal pelvis (n = 320)
0.77(0.52,1.14)
1.02 (0.72, 1.42)
0.79 (0.59, 1.07)
Note: Concentrations were log transformed and
rescaled by their SDs for analysis
Upson et al. (2013) (United States, Washington)
Population: 92 incident endometriosis cases, 195 controls
frequency-matched on age, all members of a large health
care system and enrolled in Women's Risk of Endometriosis
Study, 1996-2001; ages 18-49 yrs
Outcome: Endometriosis confirmed by surgery; for each
case, reference date assigned by date of first visit for
symptoms leading to diagnosis; reference dates randomly
assigned to controls based on case distribution
Exposure: Urine sample, collected after enrollment
(2001-2002)
MBzP in urine, controls:
Median (interquartile range)
Unadjusted (ng/mL) 5.0 (2.0-11.5)
Analysis: Logistic regression (quartiles of exposure),
covariates considered based on directed acyclic graph; final
model adjusted for variables shown in results column
OR (95% Cl) for endometriosis by quartile MzBP
(adjusted for In-transformed urinary creatinine,
age, and reference year)
MBzP quartile (ng/mL)
1 (<2.0)
2 (2.0-4.0)
3 (5.0-11.5)
4 (>11.5)
(trend p-value)
OR (95% Cl)
1.0 (referent)
1.7 (0.8, 3.8)
1.5 (0.6, 4.0)
1.3 (0.4, 4.0)
(0.80)
Adjustment for education, smoking status, and
alcohol consumption did not alter the results
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Huang et al. (2010) (Taiwan)
Population: 28 endometriosis cases, 36 leiomyoma cases,
n = 16 adenomyosis cases, n = 29 controls recruited from
the laparotomy patients in medical center, 2005-2007;
mean ages ~38, 41, and 36 yrs, respectively
Outcome: Clinical diagnosis of endometriosis, leiomyoma,
or adenomyosis confirmed by pathology
Exposure: Urine sample
MBzP in urine, controls:
Median (range)
Unadjusted (ng/mL) 5.9 (2.1-26.2)
Cr-adjusted (u.g/g Cr) 8.9 (2.1-38.7)
Analysis: Logistic regression, considering age, BMI, and
GSTM1 polymorphism as covariates
OR (95% Cl) for case status by MBzP above
compared with below the median (for
endometriosis, adjusted for GSTM1 polymorphism
and BMI; for leiomyomas and adenomyosis,
adjusted for GSTM1 polymorphism and age)
Endometriosis Leiomyomata Adenomyosis
1.07
(0.35, 3.28)
1.40
(0.48, 4.05)
1.33
(0.29,6.13)
Weuve et al. (2010) (United States, NHANES)
Population: 87 endometriosis cases, 151 leiomyomata
cases, 1,020 controls from population-based survey
(NHANES), 1999-2004; ages 20-54 yrs, mean age ~36 yrs
Outcome: Self-reported diagnosis of endometriosis or
leiomyomata; median time since diagnosis, 9 yrs
Exposure: Urine sample, collected at time of survey
MBzP in urine, controls:
Geometric mean (SE)
Cr-adjusted (ng/mg Cr) 14.1(0.6)
Analysis: Logistic regression, adjusting for variables shown
in results column
OR (95% Cl) for gynecological condition by quartile
of MBzP (ng/mg Cr) (adjusted for age, race/
ethnicity, age at menarche, current pregnancy
status and current breast-feeding status)
MBzP
quartile
1 (low)
2
3
4 (high)
(trend p)
Endometriosis
1.0 (referent)
0.84 (0.37, 1.89)
1.17(0.47,2.94)
1.17(0.42,3.27)
(0.6)
Leiomyomata
1.0 (referent)
1.11(0.59,2.07)
1.16(0.64,2.13)
1.14(0.54,2.39)
(0.8)
Itoh et al. (2009) (Japan)
Population: 57 endometriosis cases, 80 controls; all seeking
evaluation for infertility
Outcome: Clinical diagnosis of endometriosis (American
Fertility Society stages II-IV) by laparoscopy; controls were
stages 0-1
Exposure: Urine sample
MBzP in urine, controls:
Median 75th percentile
Unadjusted (u.g/L) 3.2 6.5
Cr-adjusted (u.g/g Cr) 1.8 3.3
Analysis: Logistic regression, adjusting for variables shown
in the results column
OR for endometriosis by MBzP (u.g/g Cr), above
compared with below the median (adjusted for
menstrual regularity and average menstrual cycle
length)
OR (95% Cl) = 1.38 (0.65, 2.91)
Median MBzP in urine by stage of endometriosis
Endometriosis
stage
0
I
IV
(trend p)
Unadjusted
(ug/U
3.0
3.7
4.6
3.3
4.4
(0.06)
Cr-adjusted
Cr)
1.8
1.9
2.9
2.0
2.0
(0.37)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Reddvetal. (2006a) (India)
Population: 49 endometriosis cases, 38 gynecology patient
controls (group 1), 21 tubal sterilization controls (group 2),
time period not reported; mean age ~27 yrs
Outcome: Endometriosis based on laparoscopy (American
Fertility Society severity staging)
Exposure: Plasma sample
BBP in plasma (u.g/mL):
Mean ±SD
Control group 1 0.12 ±0.20
Control group 2 0.11 ±0.22
Analysis: Two-sample t-test for comparisons between
groups; correlation analysis for association with severity
(details not reported)
Plasma BBP, mean ± SD, |Jg/mL
Control 1 Control 2 Endometriosis
0.12 ±0.20 0.11 ±0.22 0.66 ±0.61
p < 0.0002 compared with either control group
BBP concentration positively correlated with
severity (r = 0.73, p <0.0001)
Reddvetal. (2006b) (India)
Population: 85 endometriosis cases, 135 tubal sterilization
controls, from subfertility clinic, 1999-2005; mean age
~31yrs
Outcome: Endometriosis based on laparoscopy (American
Fertility Society severity staging)
Exposure: Plasma sample
BBP in plasma (u.g/mL):
Mean ±SD
Controls 0.14 ±0.26
Analysis: ANOVAfor concentration comparisons across
stages
Plasma BBP, mean ± SD (u.g/mL), by stage of
endometriosis
Controls
Stage I
Stage II
Stage III
Stage IV
0.14 ±0.26
0.28 ±0.38
0.67 ±0.50
0.98 ±0.59
1.27 ±0.61
p <0.05 for difference between means
Polycystic ovarian syndrome
Hart et al. (2013) (Australia)
Population: 121 girls from birth cohort study (Western
Australian Pregnancy Cohort), whose mothers were
recruited at 18 wks of gestation between 1989 and 1991;
follow-up at ages 14-16 yrs
Correlation coefficient (p-value) between log-
transformed MBzP and parameter
Uterine volume (mL)
Ovarian volume (cm3)
Antral follicle count
r<0.20(p>0.17)
r<0.10(p>0.29)
r < -0.01 (p >0.25)
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Outcome: Uterine volume, ovarian volume, and antral
follicle count measured by ultrasound; PCO defined as
>1 ovary more than 10 cm3 or >12 follicles between 2 and
9 mm in diameter; PCOS defined either as (1) presence of at
least two of: polycystic ovarian morphology, clinical or
biochemical hyperandrogenism, or oligo-anovulation; or
(2) oligo-anovulatory menstrual cycles with either clinical or
biochemical hyperandrogenism; all clinical assessments
conducted on d 2-5 of menstrual cycle
Exposure: Maternal serum samples (n = 123) collected at
18 and 34-36 wks of gestation (combined aliquot from both
time periods)
MBzP in serum (ng/mL):
Median 90th percentile
MBzP 1.26 3.87
Analysis: Correlation between log-transformed MBzP and
uterine volume, ovarian volume, and antral follicle counts;
MBzP concentrations in PCO or PCOS cases and controls
compared calculated using t-tests or Mann-Whitney U-tests
Authors reported no association between MBzP
and polycystic ovarian syndrome using either
definition (quantitative results not reported)
1
2
3
NHANES = National Health and Nutrition Examination Survey; PCO = polycystic ovarian morphology;
PCOS = polycystic ovarian syndrome
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.2.8. Pregnancy Related Outcomes
2 Table 3-8. Evidence pertaining to BBP and pregnancy outcomes in humans
Reference and study design
Results
Fetal growth (birth weight, birth length, head circumference)
Huang etal. (2014b) (China)
Population: 207 women delivering at one hospital in
Chongqing between 2011 and 2012, aged 18-35 yrs,
with no history of tobacco or alcohol use; mean age
28 yrs
Outcome: Standard clinical measures at birth
Exposure: Cord blood sample
BBP in cord blood (ug/L):
Median 75th percentile 95th percentile
All samples (57.2)
(trend p-
value)
In (MBzP)
Birth weight Birth
0 (referent)
14
(-141,170)
-50
(-223, 123)
(0.43)
-23
(-71, 24)
length (cm)
0
(referent)
0.0
(-0.7, 0.7)
0.1
(-0.9, 0.7)
(0.88)
0.1
(-0.3, 0.2)
Head
circumference
(cm)
0 (referent)
-0.2
(-0.8, 0.4)
-0.3
(-0.9, 0.3)
(0.32)
0.0
(-0.2, 0.2)
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Suzuki etal. (2010) (Japan)
Population: 149 infants from birth cohort,
2005-2008
Outcome: Standard clinical measurements at birth
Exposure: Maternal urine sample, gestation wks
9-40 (mean ± SD = 29 ± 8 wks)
MBzP in urine:
Median 75th percentile
Unadjusted (ng/mL) 3.46 11.2
Cr-adjusted (mg/g Cr) 4.70 9.83
Analysis: Pearson's correlation analysis for
individual metabolites and high MW phthalates
(IMBzP, MEHP, MEHHP, and MEOHP molar
concentration)
Pearson's correlation coefficient between MBzP (mg/g Cr)
or high MW phthalate (molar concentration) and birth
outcome
High MW
phthalate (molar
Birth outcome MBzP (mg/g Cr) concentration)
Birth weight (g) 0.005
Birth length (cm) -0.030
Head -0.113
circumference (cm)
Gestational age 0.069
(wks)
p >0.5 for all correlations
-0.096
-0.064
-0.072
0.043
Wolff et al. (2008) (United States, New York City)
Population: 382 singleton live births without
medical complications from birth cohort (Mt. Sinai
Children's Environmental Health study), 1998-2002
Outcome: Standard clinical measurements at birth
Exposure: Maternal urine sample, third trimester
MBzP in urine (ng/mL):
Median 75th percentile
Unadjusted 22 50
Analysis: Linear regression, adjusting for variables
shown in results column
Regression coefficient (95% Cl) for change in birth
outcome with unit increase in In-MBzP (ng/mL) (adjusted
for race/ethnicity, infant sex, gestational age at delivery,
In-creatinine, prenatal smoking, prepregnancy BMI,
maternal education, and marital status)
Birth weight (g)
Birth length (cm)
Head circumference (cm)
1.4 (-34, 37)
0.20 (0.00, 0.40)
0.11 (-0.02, 0.25)
Restricted to observations with creatinine >20 mg/dL
Preterm birth (<37 wks) and gestational age
(Ferguson et al. (2014a): Ferguson et al. (2014c))
(United States; Boston)
Population: 130 cases, 352 controls from pregnancy
cohort (study of predictors of pre-eclampsia,
enrolled first trimester), 2006-2008; controls
randomly selected from those delivering >37 wks of
gestation; mean age 33 yrs
Outcome: Preterm birth (<37 wks of gestation;
gestational age estimated from first trimester
ultrasound)
Exposure: Maternal urine samples (one to four
samples at median 9.7,17.9, 26.0, and 35.1 wks of
gestation; last sampling period not included for
mothers who had already delivered)
SG-adjusted MBzP in urine (u.g/L), geometric mean
of visits 1-3:
Geometric mean 75th percentile
Controls 6.34 10.9
Cases 6.85 13.4
Analysis: Logistic regression, considering maternal
age, race/ethnicity, education level, health insurance
OR (95% Cl) for preterm birth per unit increase in
In-transformed MBzP (geometric mean of visits 1-3)
(adjusted for urine specific gravity, maternal age,
race/ethnicity, education level, and insurance provider
(Ferguson etal., 2014a)
All preterm
Spontaneous
preterm
1.09 (0.86,1.38)
1.41(1.02,1.95)
[Results weaker than those seen with DEHP metabolites]
OR (95% Cl) for preterm birth per unit increase in
In-transformed MBzP at each study visit (adjusted for urine
specific gravity, maternal age, race/ethnicity, education
level, and insurance provider) (Ferguson et al., 2014c)
Visit 1
Visit 2
Visit 3
Visit 4
1.02 (0.73,1.43)
1.07 (0.73, 1.55)
1.00 (0.68, 1.48)
1.02 (0.57, 1.84)
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
provider, BMI at first study visit, smoking status,
alcohol use, parity, use of assisted-reproductive
technology, and sex of infant as potential covariates;
additional analyses conducted for subgroup with
preterm labor or premature rupture of membranes
("spontaneous preterm," n = 57)
Ferguson et al. (2014c) provides the analysis based
on individual sample results for each of the four
visits
Huang etal. (2014b) (China)
Population: 207 women delivering at 1 hospital in
Chongqing between 2011 and 2012; aged 18-35 yrs
and with no history of tobacco or alcohol use; mean
age 28 yrs
Outcome: Preterm birth (<37 wks of gestation;
gestational age estimated from last menstrual
period)
Exposure: Cord blood sample
BBP in cord blood (u.g/L):
Median 75th percentile 95th percentile
All samples (
-------
Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Outcome: Preterm birth (<37 wks of gestation),
determined using maternal recall of last menstrual
period
Exposure: Maternal urine sample, third trimester
MBzP in urine, among term births
Median 75th percentile
Unadjusted 2.9 5.2
SG-adjusted (jjg/L) 3.2 7.8
Cr-adjusted (ng/g Cr) 4.6 9.1
Analysis: Logistic regression, considering maternal
age, pre-pregnancy BMI, parity, education, marital
status, infant's sex, and gestational age at urine
sample as potential covariates
Cr-unadjusted (ng/L)
SG-adjusted (ng/L)
Cr-adjusted (ng/g Cr)
2.5 (0.8, 8.5)
2.2 (0.7, 6.7)
2.2 (0.7, 6.7)
Wolff et al. (2008) (United States, New York City)
Population: 382 singleton live births without
medical complications from birth cohort (Mt. Sinai
Children's Environmental Health study), 1998-2002
Outcome: Standard clinical measurements at birth
Exposure: Maternal urine sample, third trimester
MBzP in urine (ng/mL):
Median 75th percentile
Unadjusted 22 50
Analysis: Linear regression, adjusting for variables
shown in results column
Regression coefficient (95% Cl) for change in gestational
age with unit increase in In-MBzP (ng/mL) (adjusted for
race/ethnicity, infant sex, gestational age at delivery,
In-creatinine, prenatal smoking, pre-pregnancy BMI,
maternal education, and marital status)
Gestational age (wks) 0.07 (-0.07, 0.22)
Restricted to observations with creatinine >20 mg/dL
Early pregnancy loss
Toft et al. (2012) (Denmark)
Population: 48 women with pregnancy loss, 80 with
pregnancies ending in a live birth from cohort of
couples planning first pregnancy, 1992-1994
Outcome: Any pregnancy loss (n = 48), early
(subclinical) embryonal loss (pregnancy identified by
elevation in human chorionic gonadotropin; n = 32)
or clinically-identified pregnancy loss (n = 16)
Exposure: Urine samples (one conception cycle, one
preconception cycle)
MBzP in urine (ng/mL), among live births:
Mean Maximum
Live birth 20.3 117
Analysis: Logistic regression, adjusting for variables
shown in results column
OR (95% Cl) for any pregnancy loss by tertile MBzP (ng/mL)
in the preconception cycle or conception cycle (adjusted
for age, BMI, smoking, alcohol and caffeine intake, and
MBzP in the other cycle)
MBzP Tertile
1 (low)
2
3 (high)
Preconception
cycle
1.0 (referent)
1.38(0.53,3.62)
0.59 (0.21, 1.65)
Conception cycle
1.0 (referent)
1.72 (0.63, 4.69)
2.10(0.74,5.88)
OR (95% Cl) for types of pregnancy loss by tertile MBzP
(ng/mL) in the conception cycle (adjusted for age, BMI,
smoking, alcohol and caffeine intake, and MBzP in the
preconception cycle)
Subclinical Clinical pregnancy
MBzP tertile pregnancy loss loss
1 (low) 1.0 (referent) 1.0 (referent)
2 2.39 (0.70, 8.22) 1.08 (0.25, 4.66)
3 (high) 3.11(0.87,11.09) 0.96(0.20,4.59)
1 MEHHP = mono-(2-ethyl-5-hydroxyhexyl)phthalate; MEHP = mono-(2-ethylhexyl) phthalate; MEOHP = mono-
2 (2-ethyl-5-oxohexyl) phthalate; MW = molecular weight
This document is a draft for review purposes only and does not constitute Agency policy.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.2.9. Immune Effects in Humans
2 Table 3-9. Evidence pertaining to BBP and allergy/immune effects in humans
Reference and study design
Results
Ait Bamai et al. (2014) (Japan)3
Population: Children (n = 122, ages
<15 yrs) and adults (n = 374, ages
>15 yrs) living in 148 detached dwellings
in which at least 25 mg of dust was
collected; 2006 follow-up of 2003
baseline survey
Outcome: Allergic condition assessed by
self-administered questionnaire (positive
response to: in the past 2 yrs have you
been seen at a hospital for allergic
rhinitis, allergic conjunctivitis, or atopic
dermatitis?); parents completed
questionnaires for children <6 yrs old)
Exposure: Dust samples
BBP in dust (u.g/g dust) (percentile):
Median 75th
Floor dust (n = 148) 1.9 3.9
Multi-surface dust (n = 120) 1.7 3.9
Analysis: Generalized linear mixed effects
model, considering gender, age strata
(<15, >15 yrs, smoking status (personal
and environmental tobacco smoke), furry
pets in home, signs of dampness, Der 1
(not defined by authors), other
phthalates dust, airborne fungi,
formaldehyde, total VOC, and building
characteristics as potential covariates
OR (95% Cl) for allergic condition by tertile of BBP in floor dust (u.g/g
dust) (adjusted for gender, age strata, smoking status, dampness
index, furry pets inside the home, Der 1, and sum of other phthalates)
BBP tertile Full sample Children Adults
Allergic rhinitis
1 (low) 1.0 (referent) 1.0 (referent) 1.0 (referent)
2 1.27(0.64,2.52) 1.90(0.63,5.75) 0.85(0.39,
1.86)
3 (high) 1.98 (0.98, 4.03) 3.04 (0.92, 10.0) 1.29 (0.60,
2.80)
(trend p-value) (0.058) (0.068) (0.51)
Allergic conjunctivitis
1 (low) 1.0 (referent) 1.0 (referent) 1.0 (referent)
2 0.65(0.23,1.83) 0.66(0.14,3.12) 0.64(0.19,
2.17)
3 (high) 1.40(0.56,3.49) 1.48(0.33,3.56) 1.34(0.47,
3.79)
(trend p-value) (0.46) (0.61) (0.59)
Atopic dermatitis
1 (low) 1.0 (referent) 1.0 (referent) 1.0 (referent)
2 3.69 (1.41, 9.68) 4.02 (1.01, 14.5) 3.40 (0.78,
14.8)
3 (high) 5.46(2.06, 6.55(1.71,25.3) 4.54(1.06,
14.48) 19.4)
(trend p-value) (0.001) (0.007)
p-value for age interaction >0.2 for all endpoints
(0.041)
Results for multisurface dust stratified by age indicated increased
adjusted ORs for allergic conjuctivitis in children; ORs (95% Cl) were
9.81 (1.06, 91.23) in BBP tertile 2 and 7.71 (0.85, 69.97) in tertile 3.
No other significantly increased adjusted ORs (either in the full sample
or stratified by age) were observed in analyses using BBP
measurements in multisurface dust.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Wang et al. (2014) (Taiwan)
Population: 218 children from birth
cohort, born 2004-2005; follow-up at
age 2 (n = 218) and age 5 (n = 191)
Outcome: Atopic dermatitis based on
ISAAC questionnaire (three questions-
itchy rash coming and going for at least
6 mo; if yes, itchy rash in last 12 mo; ever
diagnosed with atopic dermatitis by a
doctor?); total serum IgE
Exposure: Maternal urine sample, third
trimester; urine samples in children (ages
2 and 5 yrs)
Cr-adjusted MBzP in urine (ng/g Cr):
Geometric mean (SE)
At 3rd trimester 1.84(1.11)
Age 2 3.76(1.10)
Age 5 3.46 (1.08)
Analysis: Linear regression and logistic
regression of log transformed data,
considering sex, gestational age, parity,
maternal age, education and occupation,
diets and supplements during pregnancy,
family income, parental atopy, duration
of breast feeding, tobacco smoke
exposure, incense and carpets in home,
and fungi on house walls as potential
covariates
OR (95% Cl) for atopic dermatitis by quartile of MBzP (ng/g Cr)
(adjusted for gender, gestational age, maternal education, maternal
history of atopy, and prenatal environmental tobacco smoke
exposure)
MBzP quartile (|Jg/g
Cr)
1 (<1.9048)
2 (1.9048-4.4776)
3 (4.4776-8.2000)
4 (>8.2000)
Age 2 yrs
1.0 (referent)
1.10(0.44-2.69)
0.93 (0.37-2.31)
2.50(1.08-5.79)
Age 5 yrs
1.0 (referent)
1.12(0.43-2.88)
0.59 (0.22-1.61)
1.98 (0.81-4.87)
Regression coefficient (p-value) for log-serum total IgE at 2 yrs of age
according to log-urine phthalate metabolite concentrations at age
2 yrs (adjusted for gestational age, maternal education, maternal
history of atopy, and prenatal environmental tobacco smoke
exposure)
All children (n = 218)
Boys(n = 114)
Girls (n = 104)
0.008 (0.84)
0.016 (0.74)
-0.006 (0.96)
(Callesen et al. (2014a): Callesen et al.
(2014b)) (Denmark)3
Population: 81 rhinoconjunctivitis cases,
88 atopic dermatitis cases, and
242 healthy controls from population-
based survey (Danish Indoor
Environment and Children's Health); ages
3-5 yrs; 2008
Outcome: Allergic rhinoconjunctivitis or
atopic dermatitis based on clinical exam
and parent interview; allergic
rhinoconjunctivitis diagnosed by:
recurrence of at least two or more nasal
symptoms (pruritus, runny nose,
sneezing spells >20, nasal
stenosis/mouth breathing) and ocular
symptoms (itching, conjunctival injection,
or watery secretion in both eyes) when
exposed to allergens; atopic dermatitis
diagnosed by: presence of at least 3 of
4 major features and 3 of 23 minor
Median BBP in dust (ng/g), by case-control status based on clinical
examination, from Callesen et al. (2014a)
Home
Daycare
Weighted
average
Controls (n =
242)
3.9
15.4
7.8
Atopic
Rhinoconjunctivitis dermatitis
(n = 88)
(n = 81)
3.5
16.6
8.9
3.9
15.4
7.7
Similar results when based on case status defined by parent
questionnaire data (n = 56 rhinoconjunctivitis, n = 83 atopic
dermatitis)
OR (95% Cl) for rhinoconjunctivitis or atopic dermatitis (number of
cases and controls revised after reclassification of some cases and
controls during clinical examination and elimination of participants
with missing data on covariates) by quartile of MBzP in urine (ng/mL)
(adjusted for sex, breastfeeding <3 mo, smoking in the home, single
allergic predisposition, and social class), from Callesen et al. (2014b)
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
features; 70% of rhinoconjunctivitis and
50% of atopic dermatitis cases were IgE
positive based on 20 allergen tests
Exposure: BBP concentration in dust
samples from bedroom and daycare
centers (Callesen etal., 2014a); MBzP in
urine samples from subset of population
(76 with rhinoconjunctivitis, 81 with
atopic dermatitis, and 222 controls)
(Callesen etal., 2014b)
BBP in dust (u.g/g dust) among controls:
Median
Home 3.9
Daycare 15.4
Weighted* average 7.8
(*weighted by assumed time spent in
each environment)
MBzP in urine (ng/mL) of controls:
Median 95th percentile
Unadjusted 13.7 71.4
Analysis: Mann-Whitney U-test for
concentration comparisons between
groups; logistic regression for ORs,
considering sex, breastfeeding <3 mo,
antibiotic use, single allergic
predisposition, visible mold, visible
moisture, window condensation, cat or
dog in the home, pet avoidance, changed
cleaning habits, smoking in the home,
and social class as potential covariates
MBzP Rhinoconjunctivitis
quartile (71 cases, 216 controls)
1 (low) 1.0 (referent)
2 1.48 (0.68, 3.23)
3 0.89 (0.39, 2.01)
4 (high) 1.18 (0.56, 2.48)
Atopic dermatitis
(76 cases, 216 controls)
1.0 (referent)
1.23 (0.58, 2.63)
0.69 (0.31, 1.55)
1.43 (0.72, 2.88)
This document is a draft for review purposes only and does not constitute Agency policy,
3-30 DRAFT—DO NOT CITE OR QUOTE
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Hoppin et al. (2013)a (United States,
NHANES)
Population: 2,325 participants in
population-based survey (NHANES),
2005-2006; ages >6 yrs
Outcome: Self-administered
questionnaire current allergy symptoms
(hay fever, allergy, itchy rash, rhinitis) in
past year; allergic sensitization as
measured by serum IgE (19 allergen
specific IgEs, >0.35kU/L)
Exposure: Urine sample collected same
day as serum sample; data reported in
Unadjusted MBzP in urine (ng/L)
(percentile):
Median 75th 95th
Children 17.98 37.79 106.75
Adults 7.57 17.37 57.37
Analysis: Logistic regression, adjusting
for variables shown in results column and
sampling weights; separate analyses for
children (ages 6-17 yrs) and adults
(>17 yrs)
Prevalence and OR (95% Cl) for allergy symptoms and allergic
sensitization per unit change in log-transformed urinary MBzP level
(adjusted for age, race/ethnicity, gender, BMI, creatinine, and
cotinine)
Children (n = 779)
Hay fever (n = 3.6%
23)
Rhinitis (n = 188) 27.6%
IgE sensitization 46.1%
(any)
Adults (n = 1,546)
Hay fever (n = 7.4%
88)
Rhinitis (n = 498) 35.4%
IgE sensitization 44.0%
(any)
0.42 (0.22, 0.79)
1.02 (0.62, 1.67)
1.18(0.74,1.86)
1.68(1.09,2.59)
1.24(1.01,1.52)
1.41 (0.96, 2.06)
Authors reported that adjustment for poverty income ratio did not
alter ORs
Just et al. (2012b) (United States, New
York City)
Population: 376 children from birth
cohort (CCCEH), born 1999-2006; 376
completed at least 1 of 4 follow-ups in
yr 1 and 1 of 4 follow-ups in yr 2;
339 continued through 60 mo (4 follow-
ups between 24 and 60 mo)
Outcome: Mother's report of doctor-
diagnosis of eczema (telephone and in-
person interviews; early onset: reported
at or before age 24 mo; late onset: first
reported between 24 and 60 mo; total
serum IgE
Exposure: Maternal urine sample, third
trimester
MBzP in urine (ng/mL):
RR (95% Cl) for IQR increase in log-transformed MBzP among all
reporters (adjusted for specific gravity, sex, and race/ethnicity)
Eczema (by 24 mo)
1.52 (1.21,1.91)
OR (95% Cl) for interquartile increase in log-transformed MBzP among
consistent reporters of eczema
Eczema (by 24 mo)
Eczema (late onset)
1.91(1.23,2.97)
0.90 (0.51, 1.58)
Regression coefficient (95% Cl) for IQR increase in log MBzP
concentration and log total IgE in early onset eczema cases (adjusted
for specific gravity, sex, and race/ethnicity)
IgE (at 60 mo)
-0.14 (-0.41, 0.13)
Percentile
25th 75th
5.7 31.1
Geometric mean
Unadjusted 13.6
Analysis: Poisson and logistic regression,
considering sex, race/ethnicity, prenatal
exposure to tobacco smoke, maternal
age and education, marital status,
maternal self-report of asthma, and
maternal log total IgE as potential
covariates
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Hsu et al. (2012)a (Taiwan)
Population: 59 cases (48 with allergic
rhinitis, 36 with eczema), 42 controls,
ages 3-9 yrs, recruited through
kindergartens and day care centers,
2005-2006.
Outcome: Allergic rhinitis or eczema;
initial case/control status determined
through parent report of history; final
status determined by clinical
examination
Exposure: Settled dust samples from
child's major and minor activity rooms;
urine samples collected at clinical
examination
BBP in dust, all subjects:
Median 75th percentile
Dust (ng/g) 1.0 3.9
MBzP in urine, all subjects:
Median 75th percentile
Unadjusted (jjg/L) 4.8 11.8
Cr-adjusted (ng/g Cr) 5.1 12.9
Analysis: Logistic regression adjusting
for variables shown in the results column
OR (95% Cl) for allergic rhinitis or eczema by quartile of exposure
(adjusted for age, sex, presence of fever, medication use, parents'
smoking status, parents' allergy history, parents' education, and mo of
sampling)
BBP quartile,
dust (ng/gdust)
1 (0.08-1.00)
2 (1.00-1.00)
3 (1.01-3.88)
4 (3.89-40.16)
(trend p)
MBzP quartile,
urine (ng/g Cr)
1 (0.97-2.56)
2 (2.57-5.11)
3(5.12-12.87)
4(12.88-217.16)
(trend p)
Rhinitis
1.0 (referent)
1.0
2.04 (0.50, 8.33)
7.01(1.75,28.17)
0.006
Rhinitis
1.0 (referent)
1.27 (0.33, 4.84)
1.18(0.30,4.69)
2.31(0.55,9.70)
>0.05
Eczema
1.0 (referent)
1.0
2.00 (0.42, 9.58)
7.71(1.67,35.61)
0.011
Eczema
1.0 (referent)
2.48 (0.59, 10.50)
1.42 (0.30, 6.74)
2.27 (0.46, 11.26)
>0.05
OR for all cases (at least one among asthma, rhinitis, or eczema)
significantly elevated in highest quartile BBP in dust (OR = 5.82, 95%
Cl = 1.52, 22.32; trend p = 0.01)
Kanazawa et al. (2010) (Japan)
Population: 134 residents (41 dwellings),
including 33 reporting at least one
symptom and 101 with no reported
symptoms
Outcome: Self-reported "sick house
syndrome" symptoms (fatigue; feeling
heavy-headed; headache; nausea/
dizziness; difficulty concentrating;
itching, burning or irritation of the eyes;
irritated, stuffy, or runny nose; hoarse,
dry throat; cough; dry or flushed facial
skin; scaling/itching of the scalp or ears;
and dry, itching or red-skinned hands)
Exposure: Air and dust sample in
dwellings
BBP in room air (ng/m3):
Median Range
Total concentration <2.9 <2.9-26.6
BBP in dust (mg/kg):
Median Range
Multi-surface 2.4 <0.2-35.8
Floor 4.2 <0.2-52.1
OR (95% Cl) for mucosal symptoms per 10-fold increase in BBP
concentration (adjusted for age, gender, history of allergy, and time
spent at home; similar results with additional adjustment for moldy
odor and for condensation)
Multi-surface dust (mg/kg)
Floor dust (mg/kg)
1.9 (0.8-4.7)
1.7 (0.5-6.0)
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Analysis: Logistic regression, adjusting
for variables shown in the results column
Sun etal. (2009) (China)
Population: Cases of rhinitis (n = 89) or
eczema (n = 56) and controls (n = 331
and 118 for rhinitis and eczema analysis,
respectively), all students of Tianjin
University who had participated in a
cross-sectional study of allergic
symptoms and environmental factors;
2006-2007
Outcome: Self-reported symptoms from
questionnaire. Rhinitis = in past 12 mo,
had a problem with sneezing, or a runny,
or a blocked nose when not having a cold
or the flu, or sneezing, or a runny, or a
blocked nose, or itchy-watery eyes after
contact with furred animals or after
contact with pollen; eczema = in past
12 mo, had an itchy rash; controls
responded no to questions on
asthma/wheeze, rhinitis, and eczema
Exposure: Surface dust sample in dorm
rooms
BBP in dust (u.g/g):
Median 75th percentile
26.22 42.03
Analysis: Logistic regression for OR,
considering age, gender, passive
smoking, smoking, pet raising, atopy, and
building age as potential covariates;
Mann-Whitney U-test for comparison
between BBP concentrations of cases
and controls; t-test for comparisons
between log transformed concentrations
OR for rhinitis and eczema comparing BBP in dust (|Jg/g dust) above
and below the median (adjusted for age, gender, smoking, atopy, and
building age) reportedly did not reach statistical significance
(quantitative results not reported)
Median concentration BBP in dust (u.g/g dust)
Cases Control
Rhinitis 20.11 26.05
Eczema 19.40 22.51
p >0.35 for Mann-Whitney and t-tests
Kolarik etal. (2008) (Bulgaria)
Population: 100 cases, 77 controls from
population-based survey (ALLHOME
study), 2004-2005; ages 2-7 yrs
Outcome: Cases: positive response to
wheezing during the last 12 mo, rhinitis
during the last 12 mo, when not having a
cold, or itching rash eczema in the last
12 mo; controls: negative response to all
three questions and other questions on
history of wheezing, asthma, allergy
symptoms or diagnosis in past
Exposure: Surface dust samples from
children's bedrooms
Concentration BBP in dust (mg/g dust)
Median Mean
Controls
All cases
Rhinitis
Eczema
0.32
0.38
0.32
0.40
0.45
0.53
0.49
0.60
p-value for Dunnett test
(0.37)
(0.34)
(0.58)
(0.21)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
BBP in dust (mg/g):
Geometric mean
All homes 0.32
Analysis: Dust concentrations compared
between case and control homes overall,
and between cases with specific
symptoms in the preceding 12 mo and
controls, using Mann-Whitney U-test
(untransformed data) and Dunnett test
(log-transformed data)
Bornehag et al. (2004)a (Sweden)
Population: 198 cases, 202 controls
from population-based cohort
(Dampness in Buildings and Health
cohort) (n = 10,852), 2001-2002; ages
2-7 yrs
Outcome: Rhinitis or eczema (cases
report at least two incidents of rhinitis or
eczema in the preceding year, and at
follow-up 1.5 yrs later)
Exposure: Surface dust samples from
children's bedrooms
BBzP in dust (mg/g):
Median
All homes 0.135
Analysis: Logistic regression adjusting
for variables shown in results column
OR (95% Cl) for case status by quartile of BBP in dust (mg/g dust)
(adjusted for sex, age, smoking in home, type of building, construction
period, flooding during preceding 3 yrs, and DEHP in dust)
BBP quartile
(mg/g dust)
1 (<0.05)
2 (0.05-0.13)
3(0.13-0.25)
4 (0.25-45.55)
Rhinitis (n = 79)
1.0 (referent)
1.03 (0.44, 2.39)
1.23 (0.53, 2.88)
3.04 (1.34, 6.89)
Eczema (n = 115)
1.0 (referent)
0.84 (0.40, 1.76)
1.45 (0.71, 2.97)
2.56 (1.24, 5.32)
1
2
3
4
5
6
Additional results for this study are presented in the asthma table.
CCCEH = Columbia Center for Children's Environmental Health; ISAAC = International Study of Asthma and Allergies
in Children; IgE = immunoglobulin E; VOC = volatile organic compound
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
2
Table 3-10. Evidence pertaining to BBP and asthma/wheezing and
hypersensitivity in humans
Reference and study design
Results
Ait Bamai et al. (2014) (Japan)3
Population: Children (n = 122, ages <15 yrs)
and adults (n = 374, ages >15 yrs) living in
148 detached dwellings in which at least
25 mg of dust was collected; 2006 follow-up
of 2003 baseline survey
Outcome: Bronchial asthma assessed by self-
administered questionnaire (positive
response to: in the past 2 yrs have you been
seen at a hospital for bronchial asthma?);
parents completed questionnaires for
inhabitants <6 yrs old
Exposure: Dust samples
BBP in dust (ng/g dust) (percentile):
Median 75th
Floor dust (n = 148) 1.9 3.9
Multi-surface dust (n = 120) 1.7 3.9
Analysis: Generalized linear mixed effects
model, considering gender, age strata (<15,
>15 yrs), smoking status (personal and
environmental tobacco smoke), furry pets in
home, signs of dampness, Der 1 (not defined
by authors), other phthalates dust, airborne
fungi, formaldehyde, total VOC, and building
characteristic as potential covariates
OR (95% Cl) for bronchial asthma by tertile of BBP in floor dust
(Hg/g dust) (adjusted for gender, age strata, smoking status,
dampness index, furry pets inside the home, Der 1, and sum of
other phthalate dusts)
BBP tertile
1 (low)
2
3 (high)
(trend
p-value)
Full sample
1.0 (referent)
3.46 (0.82,
14.55)
2.97 (0.78,
11.35)
(0.11)
Children
1.0 (referent)
3.30 (0.57, 19.2)
2.98 (0.51, 17.4)
(0.23)
Adults
1.0 (referent)
3.63 (0.39,
34.2)
2.96 (0.29,
30.2)
(0.36)
p-value for age interaction = 0.95
No significantly increased adjusted ORs (either in the full sample or
stratified by age) were observed in analyses of bronchial asthma
using BBP measurements in multisurface dust
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
(Callesen et al. (2014a): Callesen et al.
(2014b))a (Denmark)
Population: 72 asthma cases, 242 healthy
controls group from population-based
survey (Danish Indoor Environment and
Children's Health); ages 3-5 yrs; 2008
Outcome: Asthma based on clinical exam
and parent interview. Asthma diagnosed by
recurrence of at least two of three
symptoms: cough, wheeze, and shortness of
breath within the previous 12 mo (symptoms
not triggered by respiratory infections); and
doctor diagnosis of asthma in combination
with ongoing treatment; 47% of asthma
cases were IgE positive based on 20 allergen
tests
Exposure: BBP concentration in dust samples
from bedroom and daycare centers (Callesen
et al., 2014a); MBzP in urine samples from
subset of population (68 with asthma and
222 controls) (Callesen etal., 2014b)
BBP in dust (ng/g), controls:
Median
Home 3.9
Daycare 15.4
Time-weighted 7.8
(weighted by assumed time spent in each
environment)
MBzP in urine (ng/mL) of controls:
Median 95th percentile
Unadjusted 13.7 71.4
Analysis: Mann-Whitney U-test for
concentration comparisons between groups;
logistic regression for ORs, considering sex,
breastfeeding <3 mo, antibiotic use, single
allergic predisposition, visible mold, visible
moisture, window condensation, cat or dog
in the home, pet avoidance, changed
cleaning habits, smoking in the home, and
social class as potential covariates
Median BBP in dust (ng/g), by case-control status based on clinical
examination, from Callesen et al. (2014a)
Home
Daycare
Time-weighted
(p >0.05 for all tests)
Controls (n = 242)
3.9
15.4
7.8
Asthma (n = 72)
2.9
18.3
8.2
Similar results when based on case status defined by parent
questionnaire data (n = 110 cases)
OR (95% Cl) for bronchial asthma (60 cases, 216 controls after
reclassification of some cases and controls during clinical
examination and elimination of participants with missing data on
covariates) by quartile of MBzP (urine sample), adjusting for sex,
breastfeeding <3 mo, smoking in the home, and single allergic
predisposition (Callesen et al., 2014b)
1 (low)
2
3
4 (high)
1.0 (referent)
1.18(0.55,2.55)
0.63 (0.26, 1.53)
1.11(0.51,2.44)
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Bertelsen et al. (2013) (Norway)
Population: 623 children from birth cohort
(Environment and Childhood Asthma study),
born 1992-1993; children with current
asthma over-sampled (follow-up
2001-2004); ages 10 yrs
Outcome: Current asthma (parental report
of history of asthma plus >1 of the following:
dyspnea, chest tightness, and/or wheezing in
previous 12 mo; use of asthma medications
in previous 12 mo; positive exercise
challenge test)
Exposure: First morning urine sample
(child's), collected at study examination
MBzP in urine (ng/L) (percentiles):
Median 75th 95th
Unadjusted 29.3 52.9 128.7
SG-adjusted 30.8 53.2 135.3
Analysis: Logistic regression, adjusting for
variables shown in the results column
OR (95% Cl) for current asthma by quartile of MBzP (ng/L)
(adjusted for urine specific gravity, sex, parental asthma, and
household income)
1: <16.9 (referent)
2: >16.9-29.2
3: >29.2-52.9
4: >52.9
1 (referent)
0.70 (0.39,1.3)
0.83 (0.45, 1.5)
1.3 (0.75, 2.4)
Increase in odds of current asthma per logio IQR MBzP
(95% Cl) = 1.2 (0.88, 1.5)
Hoppin et al. (2013)a (United States,
NHANES)
Population: 2,325 participants in
population-based survey (NHANES),
2005-2006; ages >6 yrs
Outcome: Self-administered questionnaire
(asthma, wheeze in past year)
Exposure: Urine sample collected same day
as serum sample
Unadjusted MBzP in urine (ng/L)
(percentile):
Median 75th 95th
Children 17.98 37.79 106.75
Adults 7.57 17.37 57.37
Analysis: Logistic regression, adjusting for
variables shown in results column and
sampling weights; separate analyses for
children (ages 6-17 yrs) and adults (>17 yrs)
Prevalence and OR (95% Cl) for asthma symptoms per unit change
in log-transformed urinary MBzP level (adjusted for age,
race/ethnicity, gender, BMI, creatinine, and cotinine)
Children (n = 779)
Asthma (n = 65)
Wheeze (n = 80)
Adults (n = 1,546)
Asthma (n = 116)
Wheeze (n = 219)
8.4% 1.06 (0.33,3.45)
10.7% 0.92 (0.35, 2.37)
7.4% 1.46 (1.01, 2.11)
16.6% 1.78 (1.22, 2.60)
Authors reported that adjustment for poverty income ratio did not
alter ORs
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Just et al. (2012a) (United States, New York)
Population: 244 children from birth cohort
(CCCEH), born 1999-2006; follow-up in
2006-2010, ages 4.9-9.1 yrs
Outcome: Measured feNO (1-3 measures
per child), measured seroatopy (specific IgE
to dust mite, cockroach, or mouse allergens,
>0.35 lll/mL), wheeze within past year or in
subsequent year (based on parent report at
feNO study visit and at the next study visit),
with additional information to model
wheezing phenotype
Exposure: Urine sample (child's), collected
at time of feNO measurement
MBzP in urine (ng/mL):
Geometric mean (95% Cl)
Unadjusted 24 (20-28)
Analysis: Generalized estimating equation
regression models adjusted for variables
shown in results column
Percent difference in feNO per unit increase in In-MBzP (ng/mL)
(adjusted for specific gravity, age, sex, race/ethnicity, time of day
of feNO collection, and ambient NO; similar results with additional
adjustment for seroatopy and MBP, MEP, and MEHHP)
% Difference (95% Cl)
6.8(1.1,12.9)
p- value
0.019
OR (95% Cl) for incident seroatopy (over 2-yr follow-up) by log unit
change in MBzP (adjusted for specific gravity, age at atopy
measure, time since phthalate measure, sex, and race/ethnicity)
1.08 (0.89,1.32)
OR (95% Cl) for wheeze in past year (at age 5 or 7 yrs) by log unit
change in MBzP (adjusted for specific gravity, sex, and
race/ethnicity)
Subjects in feNO
study only
Age 5 0.94 (0.72, 1.22)
n = 202
Age 7 1.12(0.81,1.54)
n = 161
Entire CCCEH
cohort
1.00 (0.83,1.22)
n = 350
1.07 (0.85, 1.34)
n = 289
Hsu et al. (2012)a (Taiwan)
Population: 9 cases, 42 controls, ages
3-9 yrs, recruited through kindergartens and
day care centers, 2005-2006.
Outcome: Initial case/control status
determined through parent report of history;
final status determined by clinical
examination.
Exposure: Settled dust samples from child's
major and minor activity rooms; urine
samples collected at clinical examination
BBP in dust, all subjects:
Median 75th percentile
Dust (ng/g) 1.0 3.9
MBzP in urine, all subjects:
Median 75th percentile
Unadjusted (ng/L) 4.8 11.8
Cr-adjusted (ng/g Cr) 5.1 12.9
Analysis: Logistic regression adjusting for
variables shown in the results column
OR (95% Cl) for asthma by quartile of exposure (adjusted for age,
sex, presence of fever, medication use, parents' smoking status,
parents' allergy history, parents' education, mo of sampling)
BBP quartile, dust (ng/g dust)
1 (0.08-1.00)
2 (1.00-1.00)
3 (1.01-3.88)
4 (3.89-40.16)
(trend p)
MBzP quartile, urine (ng/g Cr)
1 (0.97-2.56)
2 (2.57-5.11)
3(5.12-12.87)
4(12.88-217.16)
(trend p)
Asthma
1.0 (referent)
1.0
4.21 (0.35, 50.98)
3.54 (0.32, 39.06)
(>0.05)
Asthma
1.0 (referent)
Not reported
4.63 (0.15, 144.06)
68.52 (1.08, >999)
(0.03)
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Sun etal. (2009) (China)
Population: 233 cases of asthma/wheezing,
rhinitis, or eczema, and 194 controls, all
students of Tianjin University who had
participated in a cross-sectional study of
allergic symptoms and environmental
factors; 2006-2007
Outcome: Self-reported symptoms from
questionnaire. Asthma/wheezing = in past
12 months, have you had wheezing or
whistling the in the chest; have you had dry
cough at night for more than 2 wks, apart
from a cough associated with a cold or chest
infection; controls responded no to
questions on asthma/wheeze, rhinitis, and
eczema
Exposure: Surface dust sample in dorm
rooms
BBP in dust (ng/g):
Median 75th percentile
26.22 42.03
Analysis: Logistic regression for OR,
considering age, gender, passive smoking,
smoking, pet raising, atopy, and building age
as potential covariates; Mann-Whitney
U-test for comparison between BBP
concentrations of cases and controls; t-test
for comparisons between log transformed
concentrations
OR for asthma comparing BBP in dust (ng/g dust) above and below
the median (adjusted for age, gender, smoking, atopy, and building
age) reportedly did not reach statistical significance (quantitative
results not reported)
Median concentration BBP in dust (ng/g dust)
Cases
Wheezing 20.11
(p >0.5 by Mann Whitney or t-test)
Control
23.81
Bornehag et al. (2004)a (Sweden)
Population: 106 cases, 177 controls from
population-based cohort (Dampness in
Buildings and Health cohort); n = 10,852;
2001-2002; ages 2-7 yrs
Outcome: asthma/wheezing without a cold,
in the preceding year, and at follow-up
1.5 yrs later
Exposure: Surface dust samples from
children's bedrooms
BBP in dust (mg/g):
Median
All homes 0.135
Analysis: Logistic regression adjusting for
variables shown in results column
OR (95% Cl) for case status by quartile of BBP in dust (mg/g dust)
(adjusted for sex, age, smoking in home, type of building,
construction period, flooding during preceding 3 yrs, and DEHP in
dust)
BBP quartile (mg/g dust)
1 (<0.05)
2 (0.05-0.13)
3(0.13-0.25)
4 (0.25-45.55)
Asthma (n = 106)
1.0 (referent)
0.67 (0.33, 1.38)
0.88 (0.43, 1.80)
1.87 (0.92, 3.81)
1
2
3
4
5
Additional results for this study are presented in the allergy/immune table.
feNO = fractional exhaled nitric oxide; MEP = monoethyl phthalate
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.2,10. Thyroid Effects in Humans
2 Table 3-11. Evidence pertaining to BBP and thyroid hormones in humans
Reference and study design
Results
Dirtuetal. (2013) (Belgium)
Population: 152 overweight or obese adults from weight
loss cohort (ENDORUP) seen at weight management clinic,
43 age- and sex-matched controls from hospital staff and
other volunteers, enrolled 2009-2012; among obese/
overweight group, 65 received bariatric surgery and 87
received standard diet and lifestyle counseling; follow-up 3,
6, and 12 mo
Outcome: Serum thyroid hormone levels (details of blood
collection were not reported)
Exposure: Urine sample (24-hr)
MBzP in urine (ng/mL):
Median 75th percentile 90th percentile
Controls 6 11 20
Obese (at baseline) 8 16 25
Analysis: Linear regression, adjusting for variables shown
in results column
Regression coefficient (p-value) for change in
hormone level with unit change in In-MBzP
(adjusted for age, weight loss, and sex, or stratified
by sex) (0.0 = no effect)
-0.14
(0.37)
Full sample Men
Overweight/obese group
FreeT4 -0.10
(0.23)
TSH 0.11(0.19)
Referent group
FreeT4 0.30(0.06) 0.45(0.15) 0.21(0.29)
TSH 0.30(0.06) -0.11 0.44(0.02)
(0.75)
Women
-0.10(0.34)
0.03(0.83) 0.16(0.13)
Boas etal. (2010) (Denmark)
Population: 758 children from birth cohort study, born
1997-2001; examined 2006-2007, ages 4-9 yrs
Outcome: Serum thyroid hormone levels (nonfasting
sample)
Exposure: Urine sample (child's), collected same day as
serum samples
Unadjusted MBzP in urine (u.g/L):
Median 75th percentile
Boys 17 37
Girls 12 31
Cr-adjusted MBzP in urine (u.g/g Cr):
Median 75th percentile
Boys 26 49
Girls 20 42
Analysis: Linear regression, adjusting for variables shown
in the results column
Regression coefficient (p-value) for change in
hormone level with unit change in In-MBzP
(adjusted for sex and age) (0.0 = no effect)
Unadjusted MBzP
T3 -0.05 (0.016)
Free T3 -0.08 (0.032)
T4 -2.34 (0.026)
FreeT4 -0.19(0.059)
TSH -0.01 (0.47)
IGF-1 -0.01 (0.38)
IGFBP-3 -0.01 (0.27)
Cr-adjusted MBzP
-0.03 (0.27)
-0.02 (0.71)
-2.90 (0.027)
-0.32 (0.012)
0.00 (0.96)
0.00 (0.74)
0.00(0.91)
Similar patterns seen in analyses stratified by
gender. Units for hormone analyses were not
reported in the publication
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Meeker et al. (2007) (United States, Boston)
Population: 408 male partners from subfertility clinic,
2000-2004; mean (± SD) age 36 (± 5.3) yrs
Outcome: Serum thyroid hormone levels
Exposure: Urine sample, collected same day as serum
samples
MBzP in urine (ng/mL):
Median 75th percentile 95th percentile
SG-adjusted 8.16 15.7 42.4
Analysis: Linear regression, considering age, BMI, smoking
status, race, previous examination for infertility, prior
impregnation of partner, timing of blood and urine
samples, and time of day as potential covariates
Regression coefficient (95% Cl) for change in
hormone level per IQR change in SG-adjusted MBzP
(ng/mL, after back-transformation from In-MBzP)
(adjusted for age, BMI, current smoking, and time
of blood sample)
Untransformed hormone levels (0.0 = no effect)
Total T3 (ng/mL) 0.001 (-0.018, 0.021)
Free T4 (ng/dL) -0.017 (-0.046, 0.011)
Ln-transformed hormone levels (1.0 = no effect)
TSH (nlU/mL) 1.01 (0.94, 1.08)
Huang et al. (2007) (Taiwan)
Population: 76 pregnant women undergoing
amniocentesis due to age >35 yrs or abnormal a-
fetoprotein or p-hCG test, 2005-2006
Outcome: Serum thyroid hormone levels collected during
2nd trimester
Exposure: Urine sample, collected same day as serum
samples
MBzP in urine:
Median 75th percentile 95th percentile
Unadjusted (ng/mL) 0.9 0.9 33.4
Cr-adjusted (ng/g Cr) 3.7 6.0 24.0
Analysis: Spearman correlation analysis; linear regression,
adjusting for variables shown in results column
Spearman correlation coefficient between hormone
level and MBzP
Unadjusted Cr-adjusted
MBzP (ng/mL) MBzP (ng/g Cr)
T3 (ng/dL) -0.084 -0.075
T4(ng/dL) 0.034 0.040
Free T4 (ng/dL) -0.007 0.083
TSH(nlU/mL) -0.080 -0.113
All coefficients p >0.05
Adjusted regression coefficient (p-value) for change
in ln-T4 with change in In-MBzP (adjusted for age,
BMI, gestational age, and other phthalate
metabolites [MEP, MBP, MEHP, MMP])
T4 (nmole/L)
FreeT4 (pmole/L)
0.032 (0.224)
0.022 (0.232)
1
2
3
MMP = monomethyl phthalate; T3 = triiodothyronine; T4 = thyroxine; TSH = thyroid stimulating hormone
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.2,11. Pulmonary Function in Humans
2 Table 3-12. Evidence pertaining to BBP and pulmonary function in humans
Reference and study design
Results
Cakmak et al. (2014) (Canada)
Population: 3,147 participants* in
population-based survey (Canadian Health
Measures Survey), ages 6-49 yrs
Outcome: Pulmonary function based on
FVC and FEVi (expressed as percent of
values predicted based on age, height, and
sex)
Exposure: Urine sample collected at same
time as pulmonary function testing
MBzP in urine (u.g/g Cr), all participants:
Geometric mean (95%CI)
Cr-adjusted 16.4 (15.84-16.98)
Analysis: Linear regression, generalized
linear mixed models (weighted based on
sampling weights), considering BMI,
ethnicity, education, income, passive
smoking, current smoking, and ambient
conditions on day of lung function
measures (temperature, relative humidity,
barometric temperature, nitrogen dioxide,
ozone, and fine particulates (PIVh.s) as
potential covariates; stratified by age
(6-16,17-49 yrs) and sex
*Study reports number of participants
inconsistently; Table 3 reports
3,071 participants, while the Methods
section and all other data tables report
3,147 participants.
Change in pulmonary function (95% Cl) per interquartile range
increase in Cr-adjusted urinary MBzP (adjusted for age, sex,
smoking, fasting, income education, and
FEVi
FVC
FEVi/ FVC
All -0.9 (-1.6,-0.2) -0.6 (-1.2, 0.1) -0.5 (-0.9,-0.1)
participants (n
= 3,071)
By age
Age 6-16
(n = 1,642)
-0.6 (-1.5, 0.3) -0.7 (-1.6, 0.2) 0.0 (-0.5, 0.5)
Age 17-49 -0.5 (-1.5, 0.6) 0.4 (-0.6, 1.4) -0.8 (-1.4, -0.2)
(n = 1,505)
By sex
Male
(n = 1,555)
Female
(n = 1,592)
0.8 (-1.5, 3.1) 0.6 (-2.6, 3.6) -0.1 (-1.3, 1.1)
-0.3 (-2.1, 1.5) -0.6 (-2.4, 1.2) 0.1 (-0.8, 1.0)
Hoppin et al. (2004) (United States,
NHANES)
Population: 240 participants in
population-based survey (NHANES III),
1988-1994; ages 20-60 yrs
Outcome: FVC, FEVI, PEF, MMEF
Exposure: Urine sample, collected at time
of pulmonary function testing
Mean (SD) MBzP in urine:
Men Women
Unadjusted (ng/mL) 22 (3.0) 22 (2.9)
Cr-adjusted (ng/g Cr) 17 (2.5) 23 (2.4)
Analysis: Linear regression, stratified by
sex and adjusted for variables shown in
results column
Regression coefficient (SE) for change in pulmonary function
measure per interquartile range increase in MBzP (19.77 ng/g
creatinine) (adjusted for age, age squared, height, BMI, smoking,
and race)
P(SE)
FVC
FEVI
PEF
MMEF
Men
-74 (68)
-52 (56)
-226 (196)
-76(136)
Women
64 (63)
34 (54)
-153 (155)
-61 (120)
p >0.05 for all; results among nonsmokers only showed no
significant associations for either men or women
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.2,12. Neurodevelopmental Effects in Humans
2
3
Table 3-13. Evidence pertaining to BBP and neurodevelopmental effects in
humans
Reference and study design
Results
Neurobehavioral measures in school-aged children
Chopra et al. (2014) (United States, NHANES)
Population: 1,493 participants in population-
based survey (NHANES), 2001-2004, ages
6-15 yrs
Exposure: Urine sample collected same day as
NHANES exam
MBzP in urine (u.g/g Cr) (percentile):
Median 75th 95th
Cr-adjusted 24.7 48.7 96.3
Outcome: Attention deficit disorder or learning
disorder as reported by parent
Analysis: Logistic regression, considering age,
sex, race, household income, low birth weight,
health insurance coverage, routine source of
healthcare, mental health professional use in
past yr, child blood lead level, maternal age at
birth, and maternal smoking during pregnancy as
potential covariates
Geometric mean (95% Cl) Cr-adjusted MBzP in urine (ng/g Cr)
by diagnosis
Neither condition
(n = 1,262)
28.7
(26.6, 31.0)
(trend p-value = 0.14)
Attention
deficit
disorder
only (n = 56)
25.8
Learning
disorder
only
(n = 116)
28.8
Both
conditions
(n = 56)
46.6
(17.6,38.0) (22.3,37.3) (29.0,75.1)
OR (95% Cl) per 10-fold increase in Cr-adjusted log-
transformed MBzP (adjusted for sex, age, race, household
income, log-transformed blood lead, and maternal smoking
during pregnancy)
Attention deficit disorder only (n = 112) 1.5 (0.7, 3.4)
Learning disorder only (n = 173) 1.2 (0.6, 2.5)
Both conditions (n = 56) 2.0 (0.6, 6.3)
Authors reported no interaction between child's blood lead
and phthalate concentration (quantitative results not
reported)
Kobroslv et al. (2014) (United States; Minnesota,
Missouri, California, Iowa)
Population: 153 children (n = 76 girls, n = 77
boys) from birth cohort study (SFF), born
2000-2005, ages 6-10 yrs in 2010 follow-up
Outcome: Child Behavior Checklist completed by
parent
Exposure: Maternal urine sample, 3rd trimester
(mean 26.6 wks)
MBzP in urine (ng/mL):
Geometric mean (95% Cl)
Unadjusted 6.6 (5.3, 8.2)
Analysis: Linear regression, considering sex, age,
mother's education, urinary creatinine, family
stress measure, and race/ethnicity as potential
covariates
Regression coefficient (95% Cl) for change in raw score on
child behavior checklist per unit increase in In-transformed
MBzP (adjusted for sex, age, mother's education and urinary
creatinine, and family stress score)
Boys Girls
Anxiety/depression -0.06 (-0.25, 0.13) -0.20 (-0.39,-0.01)
Withdrawn 0.02 (-0.14, 0.17) -0.13 (-0.29, 0.02)
Somatic complaints 0.0 (-0.15,0.16) -0.08 (-0.24, 0.07)
Social problems* 0.06 (-0.10, 0.22) -0.14 (-0.30, 0.02)
-0.06 (-0.22, 0.11) -0.04 (-0.20, 0.12)
0.0 (-0.18, 0.19) -0.10 (-0.29, 0.08)
0.08 (-0.07, 0.23) -0.10 (-0.25, 0.05)
Thought problems
Attention problems
Rule-breaking
behavior
Aggressive
behavior
0.19 (-0.01, 0.40) 0.0 (-0.21, 0.20)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Internalizing
behavior
Externalizing
behavior
Total problems
-0.04 (-0.25, 0.18) -0.22 (-0.44, 0.0)
0.18 (-0.03, 0.40) -0.04 (-0.25, 0.17)
0.10 (-0.20, 0.40) -0.21 (-0.51, 0.10)
*Sex interaction p-value = 0.05; all other interaction p-values
>0.05
Neurobehavioral outcomes in infants and preschool-aged children
Braun et al. (2014) (United States)
Population: 175 children from birth cohort in
Ohio (HOME cohort, recruited during pregnancy,
2003-2006). Follow-up at ages 4-5 yrs
Outcome: Autistic behaviors based on Social
Responsiveness Scale completed by mother;
65 item scale, higher score = more autistic
behaviors
Exposure: Maternal urine samples, 16-26 wks of
gestation
MBzP in urine (ng/g Cr) (percentile):
Median 75th 95th
Cr-adjusted 11 17 48
Analysis: Semi-Bayesian hierarchical regression
model
Regression coefficient (95% Cl) for change in total score per
unit increase in log-transformed Cr-adjusted MBzP (adjusted
for maternal demographic and perinatal factors, depressive
symptoms, caregiving environment, and serum cotinine)
-0.8 (-2.9,1.3)
Tellez-Roio et al. (2013) (Mexico)
Population: 135 children from birth cohort
(Early Life Exposure in Mexico to Environmental
Toxicants cohort; mothers recruited during first
trimester, 1997-2003)
Outcome: Mental and psychomotor
development based on Bayley Scales of Infant
Development-ll (assessed by trained examiner,
videotaped for quality control assessment)
tested at 24, 30, and 36 mo of age
Exposure: Maternal urine sample, 3rd trimester
MBzP in urine (ng/mL):
Geometric mean (95% Cl)
SG-adjusted 3.54 (2.94, 4.26)
Analysis: Linear regression for longitudinal data,
stratified by sex and adjusted for variables
shown in results column
Related reference: Ettinger et al. (2009)
Regression coefficient (95% Cl) for change in
neurodevelopment score per unit increase in maternal In-
MBzP (adjusted for birthweight, breastfeeding practices,
weight-for-age, child's age, mother's age, mother's education,
and laboratory)
Total
sample
(n = 135)
Boys
(n = 64)
Girls (n = 71)
MDI
PDI
0.30 1.30 -0.72
(-1.11, 1.73) (-0.37, 2.97) (-2.45, 1.01)
0.10 1.79 -1.21
(-1.16,1.37) (0.14,3.45) (-3.31,0.88)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Whvattetal. (2012) (United States, New York
City)
Population: 297 children from birth cohort
(CCCEH), born 1999-2006; 3-yr follow-up, mean
age 36 mo (range 27-42 mo)
Outcome: Mental, psychomotor and behavioral
development at 3 yrs based on Bayley Scales of
Infant Development-ll (assessed by trained
examiners) and Child Behavior Checklist
(completed by parent)
Exposure: Maternal urine sample, 3rd trimester
MBzP in urine (ng/mL):
Geometric mean
Unadjusted 19.0
Analysis: Linear and logistic regression adjusting
for variables shown in results column; Wald test
used to detect sex differences
Regression coefficient (95% Cl) for change in
neurodevelopment score per unit increase in maternal In-
MBzP (adjusted for specific gravity, race/ethnicity, maternal
marital status and prenatal alcohol consumption, child's
gestational age and sex, and quality of care-taking
environment)
MDI
PDI
Boys (n = 140)
-0.45
(-2.23, 1.32)
-0.57
(-2.74, 1.60)
Girls (n = 157)
-1.07
(-2.48, 0.33)
-1.05
(-2.77, 0.67)
Adjusted OR (95% Cl) for risk of mental or psychomotor delay
(score <85) per In-unit increase in maternal In-MBzP (each
model adjusted for one or more of the following: specific
gravity, race/ethnicity, maternal marital status and prenatal
alcohol consumption, child's gestational age and sex, and
quality of care-taking environment)
Boys (n = 140) Girls (n = 157)
MDI
PDI
0.89
(0.64, 1.25)
0.96
(0.66, 1.39)
0.94
(0.66, 1.35)
1.25
(0.80, 1.95)
Regression coefficient (95% Cl) for change in neurobehavior
per unit increase in maternal In-MBzP (adjusted for specific
gravity; ethnicity; maternal IQ, demoralization, hardship,
satisfaction during pregnancy and prenatal exposure to PAH
and BPA; and child's sex and age at testing)
Boys (n = 129) Girls (n = 148)
Emotionally reactive
Anxious/depressed
Somatic complaints
Withdrawn behavior
Internalizing behavior
0.34
(-0.008, 0.69)
-0.05
(-0.46, 0.35)
-0.23
(-0.63,0.17)
0.24
(-0.09, 0.58)
0.29
(-0.83, 1.42)
0.26
(-0.05, 0.57)
0.51
(0.17,0.85)
0.42
(0.10,0.73)
0.61
(0.29, 0.93)
1.79
(0.88, 2.69)
Effect modification by gender observed for
anxious/depressed, somatic complaints, and internalizing
behavior (p-values of 0.035, 0.01, and 0.04, respectively).
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
OR (95% Cl) for child's score in the borderline or clinical range
(compared to normal) per unit increase in maternal In-MBP
(adjusted for specific gravity, maternal demoralization and
satisfaction during pregnancy, and child's sex and age at
testing)
Somatic complaints
Withdrawn behavior
Internalizing behavior
Borderline
0.83
(0.59,1.15)
0.79
(0.48, 1.28)
1.38
(1.01, 1.90)
Clinical
1.20
(0.78,1.86)
1.57
(1.07, 2.31)
1.43
(1.01, 1.90)
1
2
3
4
5
BPA = bisphenol A; FEV1= forced expiratory volume in 1 second; FVC = forced vital capacity; HOME = Health
Outcomes and Measures of the Environment; MDI = mental delay index; MMEF = maximal midexpiratory flow;
PAH = polycyclic aromatic hydrocarbon; PDI = psychomotor delay index; PEF = peak expiratory flow
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.2,13. Obesity Effects in Humans
2 Table 3-14. Evidence pertaining to BBP and obesity in humans
Reference and study design
Results
Buser et al. (2014) (United States, NHANES)
Population: Participants in population-based
survey (NHANES), 2007-2010, ages >6 yrs
[sample size not reported]
Outcome: BMI measured at exam; divided
into obese (BMI z-score >95th percentile in
children, BMI >30 in adults) and overweight
(BMI z-score 85th-95th percentiles in children,
BMI 25-29.9 in adults)
Exposure: Urine sample, collected at same
time as exam
Unadjusted MBzP in urine (ng/mL)
Geometric mean (SE):
Ages 6-19 yrs 11.94 (0.63)
Ages >20 yrs 5.88(0.25)
Analysis: Logistic regression, considering
age, race/ethnicity, sex, urinary creatinine,
poverty income ratio, calorie intake, and
serum cotinine as potential covariates in
analyses of ages 6-19 yrs; or age,
race/ethnicity, sex, education, diabetes,
alcohol consumption, cigarette smoking,
calorie intake, vigorous recreational
activities, urinary creatinine, and serum
cotinine as potential covariates in analyses of
ages >20 yrs)
OR (95% Cl) in children (6-19 yrs of age) for obesity or overweight
comparing highest quartile urinary MBzP (>27.58 ng/mL) with
lowest quartile (<5.66 ng/mL) (adjusted for age, race/ethnicity,
calorie intake, serum cotinine, urinary creatinine, and income
level)
Obese Overweight
All 2.15(0.80,5.57) 1.50(0.75,3.02)
Boys 3.99(1.20,13.23) 3.23(1.12,9.34)
Girls 0.84 (0.23, 3.06 ) 1.01 (0.45, 2.24)
OR (95% Cl) in adults (>20 yrs of age) for obesity or overweight
comparing highest quartile urinary MBzP (>143.04 ng/mL) with
lowest quartile (<2.66 ng/mL) (adjusted for age, gender,
race/ethnicity, calorie intake, recreational activity, serum cotinine,
education level, smoking status, alcohol intake, and diabetes)
All
Men
Women
Obese
1.09 (0.80,1.47)
0.97 (0.59, 1.58)
1.06 (0.61, 1.83)
Overweight
0.88 (0.64,1.21)
0.89 (0.55, 1.43)
0.78 (0.44, 1.37)
Song etal. (2014) (United States)
Population: 977 Controls from nested case-
control study of incident diabetes in the NHS
(n = 393, mean age 65.6 yrs, followed until
2010) and NHS II (n = 577, mean age 45.6 yrs,
followed until 2009)
Outcome: Change in body weight based on
self-reported data from biennial
questionnaires; self-reported body weights in
these cohorts of registered nurses was highly
accurate: a correlation coefficient of 0.96
was observed between self-reported weight
and measured weights among 184 NHS
participants
Exposure: Urine sample collected at
beginning of follow-up period (collected
2000-2001 for NHS; 1995-2000 for NHS II)
MBzP in urine (nmol/L):
Median by quartile
Unadjusted 20,47,90,252
Annual rate of weight change (95% Cl) by quartile urinary MBzP
(adjusted for cohort origin, age at sample collection, menopausal
status, smoking status, physical activity, alcohol use, alternative
healthy eating index score, caloric intake, baseline body weight,
and urinary creatinine levels)
MBzP quartile Annual rate of weight change in kg/yr (95% Cl)
(median
concentration,
nmol/L)
1 (20) 0.0 (referent)
2(47) 0.29(0.13,0.44)
3(90) 0.33(0.17,0.48)
4 (252) 0.42 (0.26, 0.57)
(trend p <0.001)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Analysis: Logistic regression, mixed-effect
models for prospective annual weight change
rate by quartile MBzP using product terms
between concentrations and year after
baseline; adjusting for variables shown in
results column
Hart et al. (2013) (Australia)
Population: 121 girls from birth cohort study
(Western Australian Pregnancy Cohort),
whose mothers were recruited at 18 wks of
gestation between 1989 and 1991; follow-up
at ages 14-16 yrs
Outcome: Offspring BMI (height and weight
measured at clinic visit on d 2-5 of menstrual
cycle)
Exposure: Maternal serum samples (n = 123)
collected at 18 and 34-36 wks of gestation
(combined aliquot from both time periods)
MBzP in serum (ng/mL):
Median 90th percentile
Unadjusted 1.26 3.87
Analysis: Correlation between log-
transformed MBzP and BMI
Authors reported no association between adolescent BMI (either
as absolute value or as age- and gender-adjusted z-score) and any
phthalate metabolite in maternal serum (r = -0.10-0.04,
p = 0.345-0.931)
Dirtuetal. (2013) (Belgium)
Population: 152 overweight or obese adults
from weight loss cohort (ENDORUP) seen at
weight management clinic, 43 age- and sex-
matched controls from hospital staff and
other volunteers, enrolled 2009-2012;
among obese/overweight group, 65 received
bariatric surgery and 87 received standard
diet and lifestyle counseling; follow-up 3, 6,
and 12 mo
Outcome: Waist circumference measured at
each follow-up visit
Exposure: Urine sample (24-hr sample)
MBzP, in urine (ng/mL) (percentile):
Median 75th 90th
Controls 6 11 20
Obese 8 16 25
(at baseline)
Analysis: Linear regression, adjusting for
variables shown in results column; treatment
of repeated urinary phthalate measures was
not specified
Regression coefficient (p-value) for change in waist circumference
with unit change in In-MBzP (adjusted for age, weight loss, and sex,
or stratified by sex) (0.0 = no effect)
Overweight/ obese
group
Referent group
Full sample
0.12(0.16)
Men
0.09 (0.56)
Women
0.08 (0.45)
-0.11(0.48) 0.08(0.77) -0.08(0.67)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Teitelbaum et al. (2012) (United States, New
York City)
Population: 387 children (80 boys, 307 girls)
in child development cohort (Growing Up
Healthy Study), 2004-2008; Hispanic and
black), 6-8 yrs at enrollment
Outcome: BMI and waist circumference
measured 1 yr after enrollment; normal
weight = BMI <85th percentile (n = 2,284);
overweight = BMI >85th percentile (n = 578)
Exposure: Urine sample, collected at
enrollment
Cr-adjusted phthalates in urine (u.g/g Cr),
median:
MBzP Ihigh MW phthalates
Boys 49.6 356.0
Girls 34.0 326.6
High molecular weight phthalate metabolites
included MECPP, MEHHP, MEOHP, MEHP,
and MBzP.
Analysis: Linear regression, considering sex,
age at baseline, sedentary hours, metabolic
equivalent hours, caloric intake, race,
ethnicity, season of urine collection, family
income, and parent education as potential
covariates; restricted to children with
creatinine >10 mg/dL
Full sample results, regression coefficient (95% Cl) for change in
body metric per unit change in In-MBzP (ng/g Cr) (adjusted for
creatinine, age, sex, sedentary hours, metabolic equivalent hours,
Hispanic ethnicity, caloric intake, season, and parental education
level)
BMI (kg/m2) -0.50 (-1.51, -0.51)
Waist circumference (cm) 0.12 (-0.91, -1.14)
Svensson et al. (2011) (Mexico)
Population: 182 women; healthy controls
without diabetes from case-control study of
breast cancer, 2007-2008; mean age 54 yrs
Outcome: BMI, waist circumference, and
waist:height ratio
Exposure: First morning urine sample
collected at time of clinical evaluation
Cr-adjusted MBzP in urine (ng/g Cr):
Geometric mean (SD)
No diabetes 7.0(2.9)
Analysis: Spearman correlation coefficient
Related references: Lopez-Carrillo et al.
(2010)
Spearman correlation coefficient between anthropometric
measure and In-MBzP in urine (ng/g Cr)
BMI (kg/m2)
Waist circumference (cm)
Waist/height ratio
(p >0.05 for all parameters)
0.0059
-0.0063
0.0883
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Hatch et al. (2008) (United States, NHANES)
Population: 4,369 (2,251 males, 2,118
females) participants in population-based
survey (NHANES), 1999-2002; ages 6-80 yrs
Outcome: BMI, waist circumference
(measured)
Exposure: Urine sample, collected at time of
obesity measurement
MBzP in urine (ng/g Cr):
Range of geometric means in different age-
sex groups = 10-35
Analysis: Linear regression, adjusting for
variables shown in results column; separate
analyses by sex-age group (ages 6-11, 12-19,
20-59, 60-80 yrs)
Results
Regression coefficient (95% Cl) for change in body metric per
quartile increase in unadjusted MBzP (ng/L), by age (age,
creatinine, height, race/ethnicity, socioeconomic status, fat intake,
dairy intake, fruit and vegetable intake, physical activity, TV/video
and computer use, and smoking status, and for women,
menopausal status, parity)
MBzP 6-11 yrs 12-19 yrs 20-59 yrs 60-80 yrs
quartile p p p p
Waist circumference, males
l(low) 1.0 1.0 1.0 1.0
(referent) (referent) (referent) (referent)
2 2.52 2.14 1.27 -0.11
(-1.71,6.74) (-0.99,5.28) (-1.34,3.87) (-3.65,3.43)
3 2.42 1.36 4.87 -1.84
(-1.43,6.27) (-1.47,4.19) (2.18,7.56) (-5.61,1.93)
4 (high) 0.55 3.10 6.63 -3.18
(-3.31,4.40) (-0.67,6.88) (3.42,9.84) (-7.64,1.29
(trend p) (0.85) (0.15) (<0.0001) (0.09)
Waist circumference, females
l(low) 1.0 1.0 1.0 1.0
(referent) (referent) (referent) (referent)
2 1.69 2.48 3.55 -1.33
(-1.63,5.02) (-0.68,5.64) (0.51,6.59) (-5.24,2.59)
3 1.33 0.59 2.08 -2.18
(-1.75, 4.41) (-2.86, 4.05) (-1.62, 5.79) (-6.26, 1.91)
4 (high) -0.50 1.46 3.18 -2.41
(-3.66, 2.66) (-3.06, 5.98) (-0.90, 7.26) (-6.65, 1.84)
(trend p) (0.65) (0.74) (0.29) (0.24)
BMI, males
l(low) 1.0 1.0 1.0 1.0
(referent) (referent) (referent) (referent)
2 1.05 0.60 0.47 -0.35
(-0.60, 2.71) (-0.65, 1.86) (-0.53, 1.48) (-1.60, 0.89)
3 1.09 0.21 1.70 -1.27
(-0.36, 2.54) (-0.85, 1.27) (0.65, 2.76) (-2.97, 0.42)
4 (high) -0.13 0.84 2.35 -1.59
(-1.53,1.28) (-0.47,2.15) (1.04,3.65) (-3.43,0.24)
(trend p) (0.80) (0.3) (0.0002) (0.06)
BMI, females
l(low) 1.0 1.0 1.0 1.0
(referent) (referent) (referent) (referent)
2 0.52 1.42 1.26 -0.67
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
4 (high)
(trend p)
(-0.89,1.92) (-0.07,2.91) (-0.11,2.62) (-2.13,0.80)
0.68 0.53 0.78 -0.72
(-0.50, 1.85) (-0.96, 2.01) (-1.01, 2.56) (-2.85, 1.40)
-0.18 0.84 0.82 -0.73
(-1.43, 1.08) (-0.97, 2.65) (-1.26, 2.90) (-2.67, 1.22)
(0.80) (0.59) (0.62) (0.49)
Stahlhut et al. (2007) (United States,
NHANES)
Population: 1,451 men in population-based
survey (NHANES), 1999-2002; ages >18 yrs;
excluded if taking insulin, oral hypoglycemic
agents, or sex hormone agonists/antagonists
Outcome: Waist circumference (measured)
Exposure: Urine sample, collected at time of
obesity measurement
MBzP in urine (ng/g Cr):
Median
Cr-adjusted 14.2
Analysis: Linear regression, adjusting for
variables shown in results column
Regression coefficient per unit increase in In-MBzP (adjusted for
age, age-squared, race/ethnicity, fat intake, calorie intake, physical
activity level, smoking exposure based on cotinine, urinary
creatinine, glomerular filtration rate, serum ALT, and GGT)
B ± SE (p-value)
Waist 1.09 ±0.36
circumference (0.005)
(n = 1,292)
Increase in waist circumference began in 3rd quartile of exposure
(data shown graphically)
1
2
3
ALT = alanine aminotransferase; GGT = gamma glutamyl transferase; NHS = Nurses' Health Study
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
2
3
3.2.14. Diabetes Effects in Humans
Table 3-15. Evidence pertaining to BBP and diabetes/insulin resistance in
humans
Reference and study design
Results
Diabetes diagnosis
Sun et al. (2014) (United States)
Population: 971 incident diabetes cases
and 970 controls from among participants
in the NHS (394 cases and 393 controls,
mean age 65.6 yrs, 2000-2008) and NHS II
(577 cases and 577 controls, mean age
45.6 yrs, 1996-2007)
Outcome: Incident type 2 diabetes assessed
in biennial follow-up questionnaires.
Confirmed based on: (a) self-report of
elevated fasting glucose >7.0 mmol/L,
random plasma glucose >11.1 mmol/L, or
plasma glucose >11.1 mmol/L and at least
one symptom (excessive thirst, polyuria,
weight loss, or hunger); (b) no symptoms
but elevated glucose on two separate
occasions; or (c) treatment with insulin or
oral hypoglycemic medication
Exposure: Urine sample, collected at
beginning of follow-up period (2000-2002
for NHS; 1996-2001 for NHS II)
Unadjusted MBzP in urine (ng//L):
Median by quartile
NHS I 3.5, 7.2, 13.4, 31.8
NHS II 8.8,17.2,33.3,87.1
Analysis: Conditional logistic regression,
adjusting for variables shown in results
column
OR (95% Cl), highest compared with lowest quartile MBzP, adjusting
for matching factors including age at sample collection, race, fasting
status, time of sample collection, menopausal status, use of
hormone replacement therapy (NHS II only), urinary creatinine
levels, BMI, smoking status, postmenopausal hormone use (NHS
only), oral contraceptive (NHS II only), physical activity, alcohol use,
family history of diabetes, history of hypercholesterolemia or
hypertension, and alternative healthy eating index score
MBzP quartile
1 (low)
2
3
4 (high)
(p-value for trend)
NHS
1 (referent)
0.91(0.55,1.51)
0.85 (0.51, 1.40)
0.82 (0.48, 1.43)
(0.54)
NHS II
1 (referent)
0.85 (0.50,1.44)
1.08 (0.62, 1.86)
1.14 (0.65, 2.01)
(0.44)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
James-Todd et al. (2012) (United States,
NHANES)
Population: 215 cases, 1,235 controls from
population-based survey (NHANES),
2001-2008; women ages 20-79 yrs
Outcome: Positive response to, "Other
than during pregnancy, have you ever been
told by a doctor or health professional that
you have diabetes or sugar diabetes?"
Exposure: Urine sample, collected at time
of survey
MBzP in urine (units not reported):
Geometric mean
Unadjusted 9.7
(based on larger sample of 2,350 women)
Analysis: Logistic regression, adjusting for
variables shown in the results column
OR (95% Cl) for diabetes by quartile of MBzP (adjusted for urinary
creatinine, age, race/ethnicity, education, poverty status, fasting
time, total caloric intake, total fat intake, smoking status, and
physical activity; little change with additional adjustment for BMI
and waist circumference)
MBzP quartile
1 (low)
2
3
4 (high)
1.0 (referent)
0.78 (0.41-1.49)
1.80(1.16-2.81)
1.96 (1.11-3.47)
Svensson et al. (2011) (Mexico)
Population: 221 women with diabetes,
182 healthy without diabetes from case-
control study of breast cancer, 2007-2008;
mean age 54 yrs
Outcome: Self-reported diabetes
Exposure: First morning urine samples
MBzP in urine (ng/g creatinine):
Geometric mean (SD)
No diabetes 7.0(2.9)
Diabetes 3.8(3.9)
Analysis: Logistic regression, adjusted for
variables shown in the results column (age
and waist-height ratio not found to be
potential confounders)
OR (95% Cl) per unit increase in In-MBzP (adjusted for creatinine
and education)
0.74 (0.55,1.00)
Markers of insulin resistance
Huang etal. (2014a) (United States,
NHANES)
Population: 3,083 participants in
population-based survey (NHANES),
2001-2008; ages 12-<80 yrs; self-reported
non-diabetic, non-pregnant participants
Outcome: Fasting blood glucose; fasting
insulin; HOMA-IR
Exposure: Urine sample at time of clinical
exam
Cr-adjusted MBzP in urine (ng/g Cr):
Median change (95% Cl) in biomarker for diabetes by quartile of
MBzP (adjusted for age, gender, race/ethnicity, fasting time, urinary
creatinine, total caloric intake, triglycerides, education, poverty,
and smoking status)
MBzP
quartile
1 (low)
2
4 (high)
Fasting glucose
referent
-0.30 (-1.48,
0.87)
-0.06 (-1.25,
1.13)
-0.24 (-1.49,
1.02)
Fasting insulin
referent
0.77(0.16,1.39)
1.09 (0.39, 1.79)
1.44 (0.50, 2.38)
HOMA-IR
referent
0.21 (0.06,
0.37)
0.26 (0.09,
0.44)
0.37(0.15,
0.59)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Median 75th percentile
Men 10.4 19.5
Women 13.4 23.8
Analysis: Logistic regression, adjusting for
variables shown in the results column
(p-value for
trend)
(0.7058)
(0.0070)
(0.0028)
Trasande et al. (2013a) (United States,
NHANES)
Population: 760 participants in the
2003-2008 NHANES, 12-19 yrs old
Outcome: HOMA-IR, calculated as fasting
glucose (mmol/L) multiplied by fasting
insulin (u.U/mL divided by 22.5
Exposure: Urine sample, collected at same
time as insulin resistance measurements.
ZHigh MW phthalates in urine (u.M):
Median 75th percentile
Unadjusted 0.50 0.98
ZHigh MW phthalates = sum of MBzP,
MCPP, MEHP, MECPP, MEHHP, MEOHP;
urinary concentration of MBzP alone not
reported
Analysis: HOMA-IR assessed as continuous
or categorical variable; categorical analysis
used cut point of 4.39, reflecting >2 SD
above the mean HOMA-IR for normal
weight adolescents with normal fasting
glucose in NHANES 1999-2002. Linear and
logistic regression analyses, adjusting for
variables shown in results column. HOMA-
IR and urinary phthalate measures natural-
log transformed for analysis.
OR (95% Cl) for insulin resistance and In-urinary metabolite
concentration (u.M), adjusted for urinary creatinine, BMI category,
continuous age, race/ethnicity, caregiver education, poverty-
income ratio, gender, serum cotinine, and caloric intake
Ln-MBzP
Ln-Zhigh MW phthalates
1.26 (0.97, 1.63)
1.45(1.13,1.87)
Regression coefficient (95% Cl) for increase in In-HOMA-IR per unit
increase in In-urinary metabolite concentration (u.M), adjusted for
urinary creatinine, BMI category, continuous age, race/ethnicity,
caregiver education, poverty-income ratio, gender, serum cotinine,
and caloric intake.
Ln-MBzP
Ln-Zhigh MW phthalates
0.02 (-0.08, 0.13)
0.26(0.13,0.40)
James-Todd et al. (2012) (United States,
NHANES)
Population: 2,092 women without history
of diabetes with various measures of insulin
resistance from population-based survey
(NHANES), 2001-2008; women age
20-79 yrs
Outcome: (Among women without history
of diabetes, fasting blood glucose (FBG)
(n = 985), HOMA-IR (n = 971), glycosolated
hemoglobin Ale (n = 2,092)
Exposure: Urine sample, collected at time
of survey
Among women without diabetes, difference (from first quartile) in
median value (95% Cl) of glucose and insulin parameters by quartile
of MBzP (Model 1 adjusted for urine creatinine, age, race/ethnicity,
education level, poverty status, fasting time, total caloric intake,
total fat intake, smoking status, and physical activity; Model 2 also
adjusted for BMI and waist circumference)
MBzP Quartile Model 1
Fasting glucose (mg/dL)
1 (low) (referent)
2 0.00 (-1.70,1.70)
3 -1.13 (-3.24, 0.98)
4 (high) -2.27 (-4.76, 0.21)
Ln (HOMA)
1 (low) (referent)
Model 2
(referent)
0.77 (-1.11, 2.64)
-1.08 (-3.34, 1.18)
-2.80 (-5.32, -0.28)
(referent)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
MBzP in urine (units not reported):
Geometric mean
Unadjusted 9.7
Analysis: Logistic regression, adjusting for
variables shown in the results column
2
3
4 (high)
Ale (%)
1 (low)
2
3
4 (high)
0.09 (-0.07, 0.25)
0.13 (-0.02, 0.28)
(referent)
0.01 (-0.04, 0.06)
0.00 (-0.05, 0.05)
-0.03 (-0.09, 0.03)
-0.01 (-0.12, 0.11)
0.06 (-0.07, 0.19)
(referent)
-0.01 (-0.05, 0.04)
-0.03 (-0.08, 0.01)
-0.03 (-0.09, 0.02)
Stahlhut et al. (2007) (United States,
NHANES)
Population: 1,451 men in population-
based survey (NHANES), 1999-2002; ages
>18yrs; excluded if taking insulin, oral
hypoglycemic agents, or sex hormone
agonists/antagonists
Outcome: HOMA-IR
Exposure: Urine sample, collected at time
of obesity measurement
MBzP in urine:
Median
Cr-adjusted (u.g/g Cr) 14.2
Analysis: Linear regression, adjusting for
variables shown in results column
Regression coefficient per unit increase in In-MBzP (adjusted for
age, age-squared, race/ethnicity, fat intake, calorie intake, physical
activity level, smoking exposure based on cotinine, urinary
creatinine, glomerular filtration rate, serum ALT, and GGT)
B ± SE (p-value)
0.061 ± 0.022 (0.005)
1
2
3
4
HOMA-IR = homeostasis model assessment of insulin resistance; MCPP = mono-(S-carboxypropyl) phthalate;
MECPP = mono(2-ethyl-5-carboxypentyl) phthalate
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
2
3
3.2,15. Cardiovascular Effects in Humans
Table 3-16. Evidence pertaining to BBP and cardiovascular disease risk
factors in humans
Reference and study design
Results
Shiue (2014) (United States, NHANES)
Population: 2,489 participants in
population-based survey (NHANES),
2011-2012; ages >20 yrs
Outcome: High blood pressure (BP) (systolic
blood pressure >140 mmHg and diastolic
blood pressure >90 mmHg)
Exposure: Urine sample collected at time of
clinical exam
MBzP in urine (units not given):
Mean ±SD
Normal BP 11.21 ± 19.74
High BP 16.91 ±29.84
Analysis: Survey-weighted logistic
regression, adjusting for variables shown in
results column; t-test for comparison
between concentrations
OR (95% Cl) for high blood pressure per unit increase in log-
transformed MBzP (adjusted for urinary creatinine, age, sex,
ethnicity, BMI, and sampling weights)
1.40(1.15,1.69)
Mean ± SD MBzP in urine (units not given) in participants with
normal and high blood pressure
Normal BP (n = 2,180)
High BP (n = 309)
*p <0.001
11.21 ±19.74
16.91 ±29.84*
Trasande et al. (2013b) (United States,
NHANES)
Population: 2,447 children in population-
based survey (NHANES), 2003-2008; ages
8-19 yrs old
Outcome: Systolic BP and diastolic BP
z-score (based on CSC norms, sex, and age);
prehypertension (BP >90th percentile for
age/height/sex); fasting serum triglycerides
(n = 906; high = >100 mg/dL); nonfasting
high density cholesterol (HDL; n = 2,555;
low = <40 mg/dL)
Exposure: Urine sample, collected at time
of BMI measurement
IHigh MW phthalates in urine (nM):
Geometric mean
BP <90th percentile 0.541
BP >90th percentile 0.509
IHigh MW phthalates = sum of MECPP,
MCPP, MEHHP, MEOHP, MEHP, and MBzP
Changes in z-score (95% Cl) per unit increase in In-phthalates
(adjusted for sex, caloric intake, television watching,
poverty:income, parental education, serum cotinine, urinary
creatinine, BMI, race/ethnicity, and age)
Systolic BP
Diastolic BP
Triglycerides
HDL
IHigh MW
phthalates
0.04 (-0.002, 0.08)
0.004 (-0.04, 0.04)
-0.28 (-2.55, 2.06)
0.42 (-0.31, 1.15)
MBzP
0.03 (-0.02, 0.08)
0.03 (-0.02, 0.09)
not reported
not reported
OR (95% Cl) for BP >90th percentile per unit increase in In-
phthalates
BP >90th percentile
High triglycerides
IHigh MW
phthalates
0.94 (0.82,1.09)
1.06 (0.90, 1.24)
MBzP
1.12 (0.87, 1.44)
not reported
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Analysis: Logistic regression for pre-
hypertension (BP >90th percentile)
classification; linear regression for systolic
BP and diastolic BP z-score and triglycerides
and HDL as continuous variable; all models
adjusted for variables shown in results
column
Results
Low HDL 0.93 (0.80, 1.07) not reported
Interactions with covariates examined in supplemental analyses;
stratified analyses showed a statistically significant association
between Ihigh MW phthalates and systolic BP for gender (males),
age (children), race/ethnicity (Hispanics), cotinine level (low and
high), and BMI (<85th percentile)
1
2
3
BP = blood pressure; HDL = high density lipoprotein
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
2
3.2,16. Cancer Effects in Humans
Table 3-17. Evidence pertaining to BBP and cancer in humans
Reference and study design
Results
Lopez-Carrillo et al. (2010) (Mexico)
Population: 233 incident cases, 221 population
controls matched by age and residency, >18 yrs
of age, >1 yr in study area, 2007-2008; mean
age 53 yrs; participation rates: 94.8% of cases
and 99.5% of controls
Outcome: Histologically-confirmed breast
cancer
Exposure: Urine sample (for cases, urine
collected on average 2 mo after diagnosis, but
before treatment)
MBzP in urine, controls:
Geometric mean
Cr-adjusted (ng/g Cr) 6.27
Analysis: Logistic regression, adjusting for
variables shown in results column
Geometric mean (95% Cl) MBzP in urine (ng/g Cr), all subjects
and by menopausal status
All
Pre-menopause
Post-menopause
*p <0.05
Controls
6.27 (5.38, 7.31)
7.22 (5.67, 9.20)
5.84(4.80,7.10)
Cases
5.43 (4.81, 6.13)
5.29* (4.42, 6.34)
5.51(4.68,6.49)
OR (95% Cl) for breast cancer, by tertile of MBzP (adjusted for
current age, age at menarche, parity, menopausal status, and
other phthalate metabolites)
MBzP tertile
(Mg/g Cr)
1(0-5.18)
2 (5.19-10.79)
3 (10.8-259)
(trend p)
Full Pre-
sample menopause
1.0 (referent) 1.0 (referent)
0.60
(0.37, 0.98)
0.46
(0.27, 0.79)
(0.008)
0.36
(0.15,0.89)
0.22
(0.08, 0.61)
(0.006)
Post-
menopause
1.0 (referent)
0.71
(0.38, 1.30)
0.61
(0.31, 1.19)
(0.169)
Urinary MBzP was inversely associated with breast cancer risk
Aschengrau etal. (1998) (United States,
Massachusetts)
Population: 261 incident cases, 753 population
controls of similar age and race; both cases and
controls were permanent residents of five Cape
Cod towns
Outcomes: Breast Cancer diagnosis
(1983-1986)
Exposure: Occupational exposure to
xenoestrogens (including phthalates) based on
self-reported full-time jobs held since age 18;
exposure determined for each job using the
NIOSH Occupational Exposure Survey Database,
chemical production and usage information,
and the expert judgment of a certified industrial
hygienist
Analysis: Logistic regression, adjusting for
variables shown in results column
Incidence (%) of females with probable exposure to BBP
Cases Controls
26/261 (10.0%) 100/753 (13.3%)
OR (95% Cl) for breast cancer and probable occupational
exposure to BBP (adjusted for age at diagnosis or index year,
vital status at the interview, family history of breast cancer, age
at first birth, and personal history of prior breast cancer and
benign breast disease)
Any BBP exposure
BBP-only exposure
BBP+other xenoestrogens
0.7 (0.4, 1,2)
0.9 (0.3, 2.9)
0.7 (0.4, 1.2)
3
4
NIOSH = National Institute for Occupational Safety and Health
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
l 3.3. EXPERIMENTAL STUDIES
2 3.3.1. Male Reproductive Effects
3 Table 3-18. Evidence pertaining to male reproductive puberty effects and
4 indicators of reproductive development following oral exposure to BBP
Reference and study design
Tvl et al. (2004)
Rat (CD); 30 FO and 30 Fl parental
rats/sex/dose
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
Results3
PPS or AGO (percent change compared to control)
mg/kg-day
Fl age at PPS
Fl age at PPS adjusted
for body weight
Fl neonatal AGO
F2 neonatal AGO
0 50
0 1
0 0.2
0 -2
0 0
250 750
-1 11*
-1 11*
-8* -17*
-3 -14*
Nipple retention (number per male or percentage of males)
Fl nipples (number per
male)
percent ofFl males
with at least one
nipple on PNDs 11-13
percent of Fl males
with at least one
areolae PNDs 11-13
areolae (number per
Fl)
percent ofF2 males
with at least one
nipple on PNDs 11-13
F2 number of
nipples/male
percent ofF2 males
with at least one
areola on PNDs 11-13
F2 number of areolae
0 0
0 0
3 0
0 0
0 0
0 0
2 5
0.05 0.12
0 0.72*
0 19*
1 32*
0 1.29*
0 16*
0 0.51*
5 72*
0.19 3.14*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Hotchkissetal. (2004)
Rat (Sprague-Dawley); 6 pregnant
females/dose
0, 500 mg/kg-day
Gavage
CDs 14-18
Gray et al. (2000)
Rat (Sprague-Dawley);
13-19 pregnant females/dose
0, 750 mg/kg-day
Gavage
GDsl4-PND3
Nagao et al. (2000)
Rat (Sprague-Dawley); 25/sex /dose
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
Results3
Nipple retention (number per male rat)
mg/kg-day 0 500
areolae (number per 0 1.1
Fl neonatal male)
nipples (number per Fl 0 1
adult male)
AGO (percent change compared to control)
neonatal AGDC 0% -13*
adult AGO0 0% -2
PPS or AGO (percent change compared to control)
mg/kg-day 0 750
litter mean age at PPS 0 3
AGO 0 -26*
Nipple retention (number or percentage per neonatal male)
nipples (number per 0 5.1*
neonatal male)
percent of neonatal 0 70*
males with areolae
PPS (% incidence)
incomplete PPS due to 0/19 9/46* (20%)
genital malformation
PPS or AGO (percent change compared to control)
mg/kg-day 0 20 100 500
AGO at birth 00-4 -8*
age at PPS 0 0.5 0.2 3*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Asoetal. (2005)
Rat (Crj:CD(SD)IGS); 24/sex/dose
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
Ema and Mivawaki (2002)
Rat (Wistar); 16 pregnant
females/dose
0, 250, 500, 1,000 mg/kg-day
Gavage
CDs 15-17
Ahmad et al. (2014)
Rat (Albino); PO, female (6/group)
0, 4, 20, 100 mg/kg
Gavage
GD 14 to parturition
Results3
PPS (% incidence)
mg/kg-day 0 100
Fl complete PPS 23/24 17/24
(96%) (71%)
200
22/24
(92%)
400
14/24*
(58%)
AGO (percent change compared to control)
Absolute change
Fl, AGO, males 0 1
F2, AGO, males 0 -12*
-2
-8
-3
-14*
Relative change
Fl, AGO, males 0 0
F2, AGO, males 0 -8*
-1
-8*
-2
-12*
AGO (percent change compared to control)
mg/kg-day 0 250
AGDC 0 -1
AGO adjusted for BW° 0 -4
500
-20*
-21*
1,000
-35*
-32*
AGO or developmental milestones (percent change compared to control)
mg/kg-day 0 4
20
100
AGO
Fl male AGO PND 25 0 -1
Fl male AGO PND 5 0 -6
-1
-6
-2
-8
Developmental milestones
Fl male eye opening NR NR
Fl male fur formation NR NR
Fl male pinna NR NR
detachment
Fl male testis descend - 0
NR
NR
NR
1
NR
NR
NR
0
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results3
TNO U998a)
PPS (percent change compared to control)
Rat (Wistar); PO, female (28/group)
0,100,1,000, 3,000 u.g/L (equivalent
to 0.016, 0.171, 0.489 mg/kg-day,
average of reported intake over
premating, gestation, and lactation)
Drinking water
FO females: 2 weeks prior to mating,
through mating, gestation, and
lactation; FO males: during mating;
Fl animals were not treated after
weaning
mg/kg-day
0.015
0.147
0.442
Fl male PPS
NR
NR
NR
NR
1
2
3
4
5
6
7
8
9
*Statistically different from controls (p <0.05) as reported by study authors.
aPercent change compared to control calculated as 100 x ((treated value - control value) 4- control value).
b Calculated as follows: [% in diet x intake food/water (mg)] -f body weight (kg) = mg/kg-day.
°Values reported by the study authors were estimated from published graphs using "Grab It!", a Microsoft Excel
based free software application used to digitizes data from image files. Publisher: www.datatrendsoftware.com.
BW = body weight; GD = gestation day; PND = postnatal day; PPS = preputial separation; NR = not reported
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
c
cr
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cT
•a
Aso etal., 2005; 2-gen; complete PPS; S-
D rats
Aso etal., 2005; 2-gen; Fl AGD; S-D rats
Aso etal., 2005; 2-gen; F2 AGD S-D rats
Hotchkiss et al., 2004; CD 14-18;
areolae number, Nipples, Adult AGD S-D
rats
Hotchkiss et al., 2004; GD14-18;
neonatal AGD S-D rats
Nagao et al., 2000; AGD at birth, age at
PPS S-D rats
Gray et al., 2000; GD 14-PND3; litter
mean age at PPS
Gray et al., 2000; GD 14-PND3; AGD,
Nipples/neonatal male, Perfect of
neonatal males with areolae,...
Tyl et al., 2004; 2-gen; Fl age at PPS F2
neonatal AGD; Fl and F2 numberof
nipples, numberofareolae
S Tyl et al., 2004; 2-gen; Fl neonatal AGD
Sumneret al., 2009; retained nipples,
areolae, decreased AGD
Ahmad, 2014; Fl male AGD PND25 and
PND5, eye opening, fur formation, pinna
detachment, and testis descent
TNO, 1998a; Fl male preputial
separation
Ema and Miyawaki, 2002; AGD, AGD
adjusted for body weight
B-BO
-mm
BB€J
= statistically significant
not statistically significant
0.01 0.1 1 10 100 100010000
Dose (mg/kg-day)
2
3
Figure 3-1. Exposure-response array of male reproductive puberty effects and
indicators of reproductive development following oral exposure to BBP.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
2
3
Table 3-19. Evidence pertaining to male reproductive toxicity following oral
exposure to BBP: Alterations in hormone concentrations, mating, and sperm
decrements
Reference and study design
Gotz et al. (2001)
Rat (Wistar (Crl:WI));
10-15 pregnant females/group
0, 10 mg/L BBP to pregnant females
during the whole pregnancy and
during lactation
Drinking water
NTP (1997b)
Rat (F344); 15 males/dose
0, 300, 2,800, 25,000 ppm
0, 20, 200, 2,200 mg/kg-day
Diet
10-week modified mating study
NTP (1997b)
Rat (F344); 15 males/dose
0, 300, 900, 2,800, 8,300,
25,000 ppm
0, 30, 60, 180, 550, "high"
mg/kg-dayb
Diet
26 weeks
Tvl et al. (2004)
Rat (CD); 30 FO and 30 Fl parental
rats/sex/dose
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-day
Diet
Multigenerational study
Results3
Mating behavior (percent change compared to control)
mg/L 0 10
% male-typical 0 -39*
mounting behavior
Sperm parameters (percent change compared to control)
mg/kg-day 0 20 200 2,200
sperm motility 0 -8 -3 Not
measured
epididymal sperm 0 -13 -30 -100*
concentration
abnormal epididymal 0 -13 -3 Not
sperm measured
Note: Percentages of motile and abnormal sperm were not measured at
the high dose due to an absence of sperm.
Sperm parameters (percent incidence)
mg/kg-day 0 30 60 180 550 'High'
seminiferous tubule 000 00 100*
hypospermia
epididymal 000 00 100*
hypospermia
Sperm count (percent change compared to control)
mg/kg-day 0 50 250 750
Fl epididymal sperm 0 4 2 -21*
count
Mating, fertility, and sperm parameters (raw percentages)
Fl mating index (%) 96.7 96.7 93.3 70.0*
Fl fertility index (%) 100 96.6 92.9 81.0*
Fl motile sperm (%) 68.6 74.0 71.7 52.1*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Hotchkissetal. (2004)
Rat (Sprague-Dawley); 6 pregnant
females/dose
0, 500 mg/kg-day
Gavage
CDs 14-18
Gray et al. (2000)
Rat (Sprague-Dawley);
13-19 pregnant females/dose
0, 750 mg/kg-day
Gavage
GD14-PND3
Nagao et al. (2000)
Rat (Sprague-Dawley); 25/sex/dose
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
Results3
Fl progressively 57.3 61.2 60.1 42.1*
motile sperm (%)
Fetal hormones (percent change compared to control)
mg/kg-day 0 500
testicular testosterone 0 -45*
production
testicular testosterone 0 -35*
concentration
fetal whole-body 0 -71*
testosterone
concentration
testicular 0 -33*
progesterone
production
Note: Data shown graphically, but magnitude of change (percent change
from control) reported by study authors in text.
Adult hormones (percent change compared to control)
mg/kg-day 0 750
adult serum 0 26
testosterone
Note: Study authors indicate that sperm production and caudal sperm
numbers were significantly affected by BBP exposure although data are not
shown.
Mating, fertility, and sperm parameters (raw percentages)
mg/kg-day 0 20 100 500
FO mating index (%) 96 96 96 100
FO fertility index (%) 91.7 83.3 95.8 96
FO sperm motility (%) 96 94 94 95
FO progressively 93 80 78 81
motile sperm (%)
Fl mating index (%) 100 94.7 90.9 91.7
Fl fertility index (%) 77.3 77.8 95 77.3
Fl adult males sperm 95 96 97 88
motility (%)
Fl adult males 83 83 85 77
progressively motile
sperm (%)
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Asoetal. (2005)
Rat (Crj:CD(SD)IGS); 24 mated pairs
of FO parents/dose
0, 100, 200, or 400 mg/kg-day
Gavage
Multigenerational study
Results3
Testosterone and sperm concentration
control)
FO serum testosterone
FO sperm
concentration
Fl weanling males
serum testosterone
Fl adult males serum
testosterone
Fl adult males sperm
concentration
0
0
0
0
0
Testosterone and sperm count (percent
mg/kg-day
FO serum testosterone
FO sperm in testes
0
0
0
(percent change compared to
3
0
33
23
-9
-23
-5
-11
8
-4
-46*
-2
0
-44*
-9
change compared to control)
100
200
Not evaluated
6
-8
400
-42
-2
Mating, fertility, and sperm parameters (raw percentages)
FO mating index (%)
FO fertility index (%)
FO epididymal sperm
motility (%)
Fl mating index (%)
Fl fertility index (%)
100
83.3
71
91.3
76.2
91.7
86.4
65
91.7
95.5
95.7
90.9
74
83.3
85
95.8
91.3
58
83.3
65
Epididymal sperm (percent incidence)
Fl spermatozoa
decreased in
epididymal lumina
0
4
8
13
Note: Spermatozoa decreased in the epididymal lumina of FO males at
400 mg/kg-day also (quantitative data not reported).
Fl sperm in testes,
caudal epididymis
epididymal sperm
motility and
abnormalities
Not affected
No treatment-related effect
authors
observed
by study
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Howdeshell et al. (2008)
Rat (Sprague-Dawley); 4-9 pregnant
females/dose
0, 100, 300, 600, 900 mg/kg-day
Gavage
CDs 8-18
Results3
Fetal testicular testosterone (percent change compared to control)
mg/kg-day 0 100 300 600
0 6 -22* -66*
900
-90*
1
2
3
4
5
6
7
8
*Statistically different from controls (p <0.05) as reported by study authors.
aPercent change compared to control calculated as 100 x ((treated value - control value) -f control value).
bThe high-dose group corresponds to 25,000 ppm BBP; a reliable estimate of dose could not be calculated. The
study authors estimated doses for all but the high-dose group based on measured body weights and food
consumption. Food consumption was not measured in the 25,000 ppm BBP group due to excessive scattering of
feed and because the mean body weight of this group was 30% lower than controls.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Gray ct at, 2000; GD 14-PND3; adult
serum T
Sumner ct a I., 2009; juvenile serum T,
testosterone
llowdcshcll et al., 2008G; D 8-18;
fetal T; S-D rats
Aso ct al., 2005; FO&F1 mating index,
fertility index; FO serum T, sperm,
cpididymal sperm motility; Fl...
Hotchkiss et al., 2004; GD 14-18;
tcsticular T & progesterone
production, testicular & fetal whole.
Nagao ct al., 2000; 2-gcn; adult Fl&FO
scrum T
Nagao ct al., 2000; 2-gcn; adult Fl&FO
mating, fertility index, sperm motility,
sperm coccnlration, Fl Weanling...
NTP, 1997b; 26wks; hypospcrmia in
seminiferous tubule & cpididymis
F344 rats
Tyl et al., 2004; 2-gcn; cpididymal
sperm count, sperm motility, mating
& fertility index CD rats
NTP, 1997b; lOwks; hypospermia in
epididymisF344 rats
Bctz, 2013 ;malc 17-beta cstradiol
Ahmad, 2014; GD 14-parturition; Fl
male daily sperm production, sperm
count, scrum T albino rats
Ahmad, 2014; GD 14-parturition; Fl
male motile sperm, sperm
abnormalities/count albino rats
TNO, 1998a; Fl male sperm
parameters
Q-B-Q
Q B-
D D D
Q B-
B-B
Q B Q
Q B •
B B—Q
0.001 0.01
0.1
• = statistically significant
:j - not statistically significant
10
100 1000 10000
Dose (mg/kg-day)
2
3
4
Figure 3-2. Exposure-response array of male reproductive toxicity following
oral exposure to BBP: alterations in hormone concentrations, mating, and
sperm decrements.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
2
3
Table 3-20. Evidence pertaining to male reproductive toxicity following oral
exposure to BBP: Histopathological changes and malformations in adults and
offspring
Reference and study design
NTP (1997b)
Rat (F344); 15 males/dose
0, 300, 2,800, 25,000 ppm
0, 20, 200, 2,200 mg/kg-day
Diet
10-week modified mating study
NTP (1997b)
Rat (F344); 15/dose
0, 300, 900, 2,800, 8,300,
25,000 ppm
0, 30, 60, 180, 550, "high"
mg/kg-daya
Diet
26 weeks
Tvl et al. (2004)
Rat (CD); 30 FO and 30 Fl parental
rats/sex/dose
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-daya
Diet
Multigenerational study
Results
Testes and epididymal histopathology (percent incidence)
mg/kg-day (FO males) 0 20 200
atrophic seminiferous 070
tubules
seminiferous tubule, 000
giant cells
seminiferous tubule 000
necrosis
epididymal 070
hypospermia
chronic inflammation 000
of epididymal tail
epididymal tail detritus 000
2,200
100*
67*
20
100*
27*
73*
Testes and epididymal histopathology (percent incidence)
mg/kg-day 0 30 60 180 550
atrophic seminiferous 00070
tubules
seminiferous tubule, 00000
giant cells
epididymal tail detritus 00070
High
100*
33*
87*
Malformations and histopathological changes (percent incidence)
mg/kg-day (Fl 0 50 250
parental males)
number of Fl 000
weanlings with at least
1 reproductive tract
malformation
percentage of Fl 000
weanlings with at least
1 reproductive tract
malformation
adult Fl testicular 10 0 14
lesions
adult Fl epididymal 1 0 11
lesions
750
25*
33*
82
54
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Hotchkissetal. (2004)
Rat (Sprague-Dawley), 6 pregnant
females/dose
0, 500 mg/kg-day
Gavage
CDs 14-18
Gray et al. (2000)
Rat (Sprague-Dawley);
13-19 pregnant females/dose
0, 750 mg/kg-day
Diet
GD14-PND3
Piersma et al. (1995)
Rat (WU); 10/sex/dose
0, 250, 500, 1,000 mg/kg-day
Gavage
Reproductive toxicity study
Results
Malformations (percent incidence)
mg/kg-day (Fl males 0
with malformations)
ventral prostate 0
seminal vesicle 0
epididymis 0
testes 0
Note: No significant effects of gestational BBP exposure on
of external or internal reproductive malformations.
500
2.9
11.8
11.8
11.8
the incidence
Malformations (percent incidence)
mg/kg-day (Fl males) 0
cleft phallus ND
hypospadias ND
vaginal pouch ND
ven tral prostate N D
agenesis
seminal vesicle ND
agenesis
epididymides agenesis ND
fluid-filled testes ND
un descen ded testes N D
absent testes ND
absent gubernacular ND
cord
Note: Data in Figure 6 shown only for exposure groups; no
controls; statistical significance not noted by study authors
750
29
29
16
27
38
67
67
22
9
57
data (ND)for
Histopathological changes (percent incidence)
mg/kg-day (FO males) 0 250 500
testicular degeneration 10 30 30
accompanied by leydig
cell hyperplasia and
appearance of cellular
debris
1,000
100*
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Nagao et al. (2000)
Rat (Sprague-Dawley); 25/sex/dose
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
Results
Histopathological changes (percent incidence)
mg/kg-day 0 20 100
500
Adult FO males
atrophic seminiferous 10 NE NE
tubules (bilateral)
epididymal cell debris 10 NE NE
(bilateral)
lymphocytic infiltration 40 NE NE
of prostate interstitium
lymphocytic/ 10 NE NE
neutrophilic infiltration
of prostate epithelium
0
0
40
10
Weanling Fl males
atrophic seminiferous 000
tubules (bilateral)
decreased 000
spermatocytes in
seminiferous tubules
(bilateral)
decreased 000
spermatogonia in
seminiferous tubules
(bilateral)
leydig cell hyperplasia 000
(bilateral)
epididymal 0 NE NE
abnormality
prostate abnormality 0 NE NE
seminal vesicle and 0 NE NE
coagulating gland
abnormality
10
90*
30
10
0
0
0
Adult Fl males
atrophic seminiferous 000
tubules (right side)
atrophic seminiferous 000
tubules (left side)
decreased germ cells in 0 0 0
seminiferous tubules
(right side)
60*
30
40*
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Asoetal. (2005)
Rat (Crj:CD(SD)IGS); 24/sex/dose
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
Results
decreased germ cells in
seminiferous tubules
(left side)
diffuse dilatation of
seminiferous tubule
(left side)
testicular interstitial
edema (right side)
testicular defect (right
side)
spermatic granuloma
of the rete testis (right
side)
multinudeated giant
cell seminiferous
tubule (left side)
epididymal lesions
lymphocytic infiltration
of prostate interstitium
lymphocytic/plasma
cell infiltration of the
prostate epithelium
0
0
0
0
0
0
0
30
20
0
0
0
0
0
0
0
NE
NE
0
0
0
0
0
0
0
NE
NE
10
10
40*
10
10
10
50*
40
20
Histopathological changes (percent incidence)
mg/kg-day (adult Fl
males)
softening of testis
aplasia ofepididymis
hypoplasia of
epididymis
leydig cell hyperplasia
atrophy of
seminiferous tubules
residue germ cells in
epididymis lumen
aplasia ofepididymis
(unilateral)
partial aplasia of
epididymis (unilateral)
partial aplasia of
epididymis (bilateral)
0
0
0
0
0
4
0
0
0
0
100
4
0
0
4
4
4
0
0
0
200
8
0
0
0
13
13
0
0
0
400
17
4
17
21*
38*
4
8
13
4
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Note: Incidence of Leydig cell hyperplasia of the testes was increased in FO
adult males in the 400 mg/kg-day group.
BIBRA (1978)
Histopathological changes (incidence)
Rat (Wistar); 27/sex/group or
45/sex/group (control); interim
sacrifices of 9 controls/sex/group
and 6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
0,151, 381, 960 mg/kg-day (males)0
0,171, 422,1,069 mg/kg-day
(females)
Diet
14 weeks
mg/kg-day
0
151
381
960
epididymis; sperm
retention cyst
histopathology
1/27
-/o
-/o
0/14
No histological lesions were noted by study authors in the prostrate,
seminal vesicles, or testis.
1
2
3
4
5
6
7
8
9
*Statistically different from controls (p <0.05) as reported by study authors.
aThe high-dose group corresponds to 25,000 ppm BBP; a reliable estimate of dose could not be calculated. The
study authors estimated doses for all but the high-dose group based on measured body weights and food
consumption. Food consumption was not measured in the 25,000 ppm BBP group due to excessive scattering of
feed and because the mean body weight of this group was 30% lower than controls.
CERHR = Center for the Evaluation of Risks to Human Reproduction; NTP = National Toxicology Program
NE = not examnied
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
NTP, 1997b; lOvvks; atrophic seminiferous
tubules, seminiferous tubule giant cells,
epididymal hypospermia, chronic tail...
NTP, 1997b; lOwks; seminiferous tubule necrosis
F3 44 rats
B1BRA, 1978; 14wks; sperm retention cyst,
prostate, seminal vesicles, ortestis histopathology
Wistarrats
NTP, 1997b; 26wks; atrophic seminiferous
tubules, seminiferous tubule giant cells,
epididymal tail detritus F344 rats
Aso etal., 2005; 2-gen; leydig cell hyperplasia,
seminiferous tubule atrophy S-D rats
Aso etal., 2005; 2-gen; softening testis,
epididymus: hypoplasiaand aplasia, seminiferous
tubule atrophy, germ cell residue S-D rats
Nagao et al., 2000; prostate, SV, S-D rats
Nagao et al., 2000; Fl weanling & adult decreased
spermatocytes; Fl adult EPI lesions, testicular
edema, atrophic seminiferous tubules S-D rats
Gray et al., 2000; GD 14-PND3; cleft phallus,
hypospadias, vaginal pouch, agensis of VP, SV, EPI;
testes fluid-filled,absent, undescended,absent...
Tyl et al., 2004; 2-gen; Fl adult testicular,
epididymal lesions CD rats
Tyl et al., 2004; 2-gen; Fl weanlings with
reproductive tract malformation CD-rats
Sumneret al., 2009; cleft phallus
Piersma etal., 1995; leydig cell hyperplasia
Hotchkiss et al., 2004; VP, SV, Epididymis, Testes
- statistically significant
= not statistically significant
o B m
B H B-
3 B-
Q B B
-B B
B B-
10 100 1000
Dose (mg/kg-day)
10000
2
3
Figure 3-3. Exposure-response array of male reproductive toxicity following
oral exposure to BBP: external and internal malformations.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
2
Table 3-21. Evidence pertaining to male reproductive toxicity following oral
exposure to BBP: Decrease in androgen-dependent tissue weights
Reference and study design
NTP (1997b)
Rat (F344); 15 males/dose
0, 300, 2,800, 25,000 ppm
0, 20, 200, 2,200 mg/kg-day
Diet
10-week modified mating study
NTP (1997b)
Rat (F344); 15 males/dose
0, 300, 900, 2,800, 8,300,
25,000 ppm
0, 30, 60, 180, 550, "high"
mg/kg-dayb
Diet
26 weeks
Tvl et al. (2004)
Rat (CD); 30 FO and 30 Fl parental
rats/sex/dose
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-day
Diet
Multigenerational study
Results3
mg/kg-day 0
20 200
2,200
Absolute weight (percent change compared to control)
ventral prostate 0
right testes 0
right epididymis 0
right cauda 0
1 -1
-8 0
-7 -10
-11 -19
-55*
-70*
-57*
-69*
Relative weight (percent change compared to control)
ventral prostate 0
right testes 0
mg/kg-day 0
1 1
-8 3
30 60 180
-36*
-58*
550 'High'
Absolute weight (percent change compared to control)
right testes 0
right epididymis 0
right cauda epididymis 0
473
3 ND ND
-13 ND ND
5 -70*
5 -47*
-7 -52*
Relative weight (percent change compared to control)
right testes 0
-2 -3 1
2 -56*
Percent change compared to control
mg/kg-day 0
50 250
750
Absolute weight (Fl males)
ventral prostate 0
seminal vesicles 0
paired testes 0
paired epididymis 0
weanling testes 0
-12 -7
1 -1
1 1
2 3
3 8*
-26*
-18*
-21*
-11*
-26*
Relative weight (Fl males)
ventral prostate 0
weanling testes 0
-13 -10
3 7*
-18*
-10*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Hotchkissetal. (2004)
Rat (Sprague-Dawley); 6 pregnant
females/dose
0, 500 mg/kg-day
Gavage
CDs 14-18
Gray et al. (2000)
Rat (Sprague-Dawley);
13-19 pregnant females/dose
0, 750 mg/kg-day
Gavage
GD14-PND3
Piersma et al. (1995)
Rat(WU);10/sex/dose
0, 250, 500, 1,000 mg/kg-day
Gavage
Reproductive toxicity study
Results3
Absolute weight (percent change compared to control)
mg/kg-day
glans penis
ventral prostate
seminal vesicles
paired testes
whole epididymis
whole cauda
epididymis
caput corpus
epididymis
LABC
Note: Tissue weight data
0
0
0
0
0
0
0
0
0
adjusted for body weights were
500
-3
-14
-6
-1
-2
-4
-4
-10*
not reported.
Absolute weight (percent change compared to control)
mg/kg-day
ventral prostate
seminal vesicles with
coagulating glands
glans penis
LABC
testes
paired epididymis
cauda epididymis
caput corpus
epididymis
Note: Tissue weight data
0
0
0
0
0
750
-42*
-38*
-19*
-34*
0 -23* (-35* reduction at
PND2)
0
0
0
-25*
-42*
-26*
adjusted for body weights not reported.
Absolute weight (percent change compared to control)
mg/kg-day
0 250 500
1,000
Adult FO males
testes and epididymis
0 -2 -2
-14*
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Nagao et al. (2000)
Rat (Sprague-Dawley); 25/sex/dose
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
Results3
Absolute weight (percent change compared to control)
mg/kg-day
0
20
100
500
Adult male FO
testes
paired epididymis
ventral prostate
Seminal vesicle
0
0
0
0
-1
-2
3
-2
3
1
7
-2
-2
-3
1
-3
Weanling male Fl offspring
testes
paired epididymis
prostate and seminal
vesicle
0
0
0
4
7
3
0
-1
-1
-12*
_9*
-9
Adult male Fl offspring (postweaning)
testes
paired epididymis
ventral prostate
seminal vesicle
0
0
0
0
0
-3
-3
-3
-3
-5
-7
-1
-12*
-21*
-14*
-10
Relative weight (percent change compared to control)
Adult male FO
testes
paired epididymis
ventral prostate
seminal vesicle
0
0
0
0
0
0
0
0
2
0
0
-3
5
5
8
3
Weanling male Fl offspring
testes
paired epididymis
0
0
2
5
-1
-2
-6*
-3
Adult male Fl offspring (postweaning)
testes
paired epididymis
ventral prostate
seminal vesicle
0
0
0
0
4
0
0
3
5
0
0
7
0
-10
-9
3
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Asoetal. (2005)
Rat (Crj:CD(SD)IGS); 24/sex/dose
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
Results3
Absolute weight (percent change compared to control)
mg/kg-day 0
100
200
400
Adult male FO
right testes 0
left testes 0
right epididymis 0
left epididymis 0
ventral prostate 0
seminal vesicle 0
5
3
3
2
-13
-2
4
4
2
2
-4
0
-1
-2
-5
-6*
-18
-7
Adult male Fl offspring (postweaning)
right testes 0
left testes 0
right epididymis 0
left epididymis 0
ventral prostate 0
seminal vesicle 0
-1
1
-1
-1
-3
-7
-7 -12*
1
-5
-9
-9
-5
-5
-17*
-16*
-13
-13*
Relative weight (percent change compared to control)
Adult male FO
right testes 0
left testes 0
right epididymis 0
left epididymis 0
ventral prostate 0
seminal vesicle 0
3
0
9
0
-15
-3
0
0
0
0
-8
-3
0
-3
0
0
-15
-9
Adult male Fl offspring (postweaning)
right testes 0
left testes 0
right epididymis 0
left epididymis 0
ventral prostate 0
seminal vesicle 0
0
0
0
0
0
-3
3
0
0
-9
-9
-7
0
-3
-9
-9
-9
-10
Autopsy findings (percent incidence)
Adult male Fl offspring (postweaning)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
TNO (1998a)
Rat (Wistar); PO, female (28/group)
0, 100, 1,000, 3,000 u.g/L (equivalent
to 0.016, 0.171, 0.489 mg/kg-day,
average of reported intake over
premating, gestation, and lactation)
Drinking water
FO females: 2 weeks prior to mating,
through mating, gestation, and
lactation; FO males: during mating;
Fl animals were not treated after
weaning
Ahmad et al. (2014)
Rat (Albino); PO, female (6/group)
0, 4, 20, 100 mg/kg
Gavage
GD 14 to parturition
Results3
small testis
small epididymis
Percent change compared to
mg/kg-day
Fl male caudal
epididymis, absolute
weight (left)
Fl male caudal
epididymis, relative
weight (left)
Fl male epididymis,
relative weight
Fl male epididymis
absolute weight
Fl male prostate,
relative weight
Fl male prostate,
absolute weight
Fl male seminal
vesicles, absolute
weight
Fl male seminal
vesicles, relative
weight
Fl male testis,
absolute weight (left)
Fl male testis, relative
weight (left)
0
0
control
0 0
0
0
0
0
0
0
0
0
0
0
0
0
015
-3
-1
0
0
-4
-4
-5
-4
0
0
0
0
0.147
-1
-1
-1
-1
-3
-2
1
0
-3
-3
25*
13
0.442
-4
3
-1
-2
-4
-4
-3
-2
-3
-3
Absolute weight (percent change compared to control)
mg/kg-day
Fl male epididymis
Fl male prostrate
Fl male seminal
vesicle
Fl male testis
0
0
0
0
0
4
-3
-2
-1
-1
20
-4
-2
-1
-1
100
-13*
-12*
-11
-2
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
BIBRA (1978)
Rat (Wistar); 27/sex/group or
45/sex/group (control); interim
sacrifices of 9 controls/sex/group
and 6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
0, 151, 381, 960 mg/kg-day (males)d
0, 171, 422, 1,069 mg/kg-day
(females)
Diet
14 weeks
Results3
Percent change compared to control
mg/kg-day 0 151
male gonad relative 0 7
weight
male gonad weight 0 -2
381 960
8 7
0 -1
1
2
3
4
5
6
7
8
9
10
11
*Statistically different from controls (p <0.05) as reported by study authors.
aPercent change compared to control calculated as 100 x ((treated value - control value) 4- control value).
bThe high-dose group corresponds to 25,000 ppm BBP; a reliable estimate of dose could not be calculated. The
study authors estimated doses for all but the high-dose group based on measured body weights and food
consumption. Food consumption was not measured in the 25,000 ppm BBP group due to excessive scattering of
feed, and because the mean body weight of this group was 30% lower than controls.
LABC = levator ani bulbocavernosus; ND = not determined
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
NTP, 1997b; lOwks; VP, testes, epididymis F344rats
B1BRA, 1978; 14wks; gonad rel and abs wtWistarrats
NTP, 1997b; 26wks; testes, epididymis F344rats
Hotchkiss et a!., 2004; GD14-18; LABCS-D rats
Hotchkiss et al., 2004; GD 14-18; VP, SV, GP, &
epididymis, testes S-D rats
Ahmad, 2014; GD 14-parturition; Fl abs epididymis
and prostate wt, albino rats
Ahmad, 2014; GD 14-parturition; Fl abs seminal
vesicle and testis wt, albino rats
Grayetal., 2000; GD14-PND3; VP,SV+CG,glans
penis, LABC, testes, epididymis
TNO, 1998a; pre-mating-lactation; Fl male tissue
weights W is tar rats
Aso etal, 2005; 2-gen; FO adult epididymis, SV, testes;
Fl adult testes S-D rats
Aso etal., 2005; 2-gen; FO&F1 adult testes, W; FO
adult SV, epididymis; FO&F1 adults testes, epididymis,
VP, SV S-D rats
Aso etal., 2005; 2-gen; abs wt Fl adult epididymis S-
D rats
Nagaoetal., 2000; FO&F1 adultVP, SV, epididymis,
testes; Fl weanling prostate&SV; Fl weanling
eididymis S-D rats
Nagao et al., 2000; 2-gen; Fl weanling & adult testes,
epididymis, Fl adult W, SV; Fl weanling testes S-D
rats
Tyl et al., 2004; 2-gen; adult VP, SV, testes, epididymis
CD rats
Tyl et al., 2004; 2-gen; weanling testes CD rats
-B—Q
ODD
D-B—B B
Q B B
B B B
D D •
B-B-B
O D •
B B B
B B B
B B •
B B-
= statistically significant
- not statistically significant
0.01 0.1 1 10 100 1000 10000
Dose (mg/kg-day)
2
3
Figure 3-4. Exposure-response array of male reproductive toxicity following
oral exposure to BBP: decrease in androgen-dependent tissue weights.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.3,2. Female Reproductive Effects
2
3
Table 3-22. Evidence pertaining to female reproductive toxicity following oral
exposure to BBP
Reference and study design
Results3
Reproductive tissue weights
Moral et al. (2007)
Rat (Sprague-Dawley CD);
10 pregnant females/dose
0, 500 mg/kg-day from PND 2 to 20
Gavage
Female offspring were evaluated at
21, 35, 50, and 100 days
Gotz et al. (2001)
Rat (Wistar (Crl:WI)); 10-15/group
0, 10 mg/L BBP to pregnant females
during the whole pregnancy and
during lactation
Drinking water
Nagao et al. (2000)
Rat (Sprague-Dawley);
20-25 breeding
pairs/group/generation; organ
weights assessed in 20-24 FO
females/group and 41-46 Fl female
weanlings/group
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
Absolute uterine weight (percent change compared to control)
mg/kg-day 0 500
day 21 0 20
day 35 0 24
day 50 0 11
day 100 0 5
Relative uterine weight (percent change compared to control)
day 21 0 27*
day 35 0 23
day 50 0 11
day 100 0 6
Absolute weight (percent change compared to control
mg/L 0 10
ovarian weight 0 -40*
Absolute weight (percent change compared to control)
mg/kg-day 0 20 100 500
FO ovaries 00-4 -11*
Fl ovaries 02-7 -16*
FO uterus 0 -8 10 18
Fl uterus 0-232
Relative weight (percent change compared to control)
FO ovaries 02-3 -11*
Fl uterus 0-44 13*
FO uterus 0 0 18 18
Fl ovaries 0 0-6-9
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Tvl et al. (2004)
Rat (CD); 30 breeding
pairs/group/generation; organ
weights assessed in 30 FO
females/group, 30 Fl adult
females/group, 67-81 Fl female
weanlings/group, and 43-87 F2
female weanlings/group
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
Asoetal. (2005)
Rat (Crj:CD(SD)IGS); 24 breeding
pairs/group/generation; organ
weights assessed in 19-20 FO
females/group and 12-19 Fl
females/group
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
Results3
Absolute weight (percent change compared to control)
mg/kg-day
FO adults ovaries
FO adult uterus
Fl weanlings ovaries
Fl weanlings uterus
Fl adults ovaries
Fl adults uterus
F2 weanlings ovaries
F2 weanlings uterus
0
0
0
0
0
0
0
0
0
50
8
-1
6
-2
3
0
0
5
250
3
3
3
4
3
9
-6
16*
750
-13*
-75*
-24*
-20*
2
20*
-19*
-14
Relative weight (percent change compared to control)
FO adults ovaries
FO adults uterus
Fl weanlings ovaries
Fl weanlings uterus
Fl adults ovaries
Fl adults uterus
F2 weanlings ovaries
F2 weanlings uterus
mg/kg-day
0
0
0
0
0
0
0
0
0
-3
-1
4
-4
2
-1
-2
4
100
-7
3
3
4
0
6
-6
13
200
-19*
-17*
-4
3
9*
28*
-8
-2
400
Absolute weight (percent change compared to control)
FO right ovary
FO left ovary
FO uterus
Fl right ovary
Fl left ovary
FO uterus
0
0
0
0
0
0
-4
0
-8
12
10
1
1
-3
-9
3
2
8
-7
-4
-4
-8
-5
12
Relative weight (percent change compared to control)
FO right ovary
FO left ovary
FO uterus
0
0
0
-4
-1
-4
-8
-8 -15*
-7
-4
-4
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1989)
Rat (Sprague-Dawley);
27-30 pregnant females/dose
0, 420, 1,100, 1,640 mg/kg-day
Diet
GDs 6-15; dams sacrificed on GD 20
NTP (1990)
Mouse (CD-I); 27-30 pregnant
females/dose (except n = 14 in the
high-dose group)
0, 182, 910, 2,330, 4,121 mg/kg-day
Diet
GDs 6-15; dams sacrificed on GD 17
Ema et al. (1998)
Rat (Wistar); 7-10 pregnant
females/dose
0, 250, 500, 750, 1,000 mg/kg-day
Gavage
GDs 0-8; dams sacrificed on GD 20
BIBRA (1978)
Rat (Wistar); 27/sex/group or
45/sex/group (control); interim
sacrifices of 9 controls/sex/group
and 6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
0, 151, 381, 960 mg/kg-day (males)"
0, 171, 422, 1,069 mg/kg-day
(females)"
Diet
14 weeks
Results3
Fl right ovary 07-3
Fl left ovary 06-3
Fl uterus 0-40
Percent change compared to control
mg/kg-day 0 420 1,100
gravid uterine weight 040
Percent change compared to control
mg/kg-day 0 182 910
gravid uterine weight 03-7
Note: The 4,121 mg/kg-day group was eliminated after evaluation
dams since all litters were completely resorbed.
Percent change compared to control
mg/kg-day 0 250 500 750
ovary weight (day 90 -2-9 -13*
pseudopregnancy)0
uterine weight (day 90 1 -34 -42*
pseudopregnancy)0
Percent change compared to control
mg/kg-day 0 171 422
female gonad relative 008
weight
female gonad absolute 008
weight
-9
-7
8
1,640
-42*
2,330
-85*
of 14
1,000
-17*
-47*
1069
-5
-8
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
TNO (1998a)
Rat (Wistar); PO, female (28/group)
0, 100, 1,000, 3,000 u.g/L
(equivalent to 0.016, 0.171,
0.489 mg/kg-day, average of
reported intake over premating,
gestation, and lactation)
Drinking water
FO females: 2 weeks prior to mating,
through mating, gestation, and
lactation; FO males: during mating;
Fl animals were not treated after
weaning
Results3
Percent change compared to control
mg/kg-day 0 0.016 0.171
Fl female absolute 06-1
ovary weight
Fl female absolute 0 -3 -2
uterus weight
Fl female relative 03-3
ovary weight
Fl female relative 0 -5 -4
uterus weight
0.489
3
1
1
0
Gross necropsy of reproductive organs
Tvl et al. (2004)
Rat (CD); 30 breeding
pairs/group/generation; gross
necropsy performed in 30 adult
females/generation
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
BIBRA (1978)
Rat (Wistar); 27/sex/group or
45/sex/group (control); interim
sacrifices of 9 controls/sex/group
and 6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
0, 151, 381, 960 mg/kg-day (males)"
0, 171, 422, 1,069 mg/kg-day
(females)b
Diet
14 weeks
Percent incidence
mg/kg-day 0 50 250
750
FO fluid-filled uterus 0300
Fl fluid-filled uterus 0 0 3 10
Note: Study authors report that the consequence of this finding is
unknown, as the affected females were in estrus at sacrifice. They also
state that increased uterine weight in Fl adults is likely due to increased
incidence of fluid filled uteri.
Response incidence
mg/kg-day 0 171 422
female uterus 0 - -
distended
female ovary No histological lesions were noted in the
histopathology
1069
1
ovaries
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results3
Puberty
Tvl et al. (2004)
Rat (CD); 30 breeding
pairs/group/generation; onset of
puberty assessed in 26-28 Fl
litters/group
0, 750, 3,750,11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
Percent change compared to control
mg/kg-day
0
50
250
750
Fl age at vaginal
opening
Fl age at vaginal
opening adjusted for
body weight
-1
-1
Moral et al. (2011)
Percent change compared to control
Rat (Sprague-Dawley CD);
10 pregnant females/dose
0,12, 500 mg BBP/kg-day from
day 10 post-conception to delivery
Gavage
Litters were euthanized at 21, 35,
50 and 100 days
mg/kg-day
0
120
500
day of vaginal
opening
body weight (g)
at day of vaginal
opening
-1
Moral et al. (2007)
Development of the mammary gland (number of terminal end buds)
Rat (Sprague-Dawley CD);
10 pregnant females/dose
0, 500 mg/kg-day from PND 2 to 20
Gavage
Female offspring were evaluated at
21, 35, 50, and 100 days
mg/kg-day
0
500
day 21
day 35
day 50
day 100
0
0
0
-1
24
82
Development of the mammary gland (number of terminal ducts)
day 21 06
day 35 0 -12
day 50 0 13
day 100 0 8
Development of the mammary gland (number of alveolar buds)
day 21 0 -29
day 35 0 -8
day 50 0 -12
day 100 0 0
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Nagao et al. (2000)
Rat (Sprague-Dawley):,
20-25 breeding
pairs/group/generation; onset of
puberty assessed in 39-48 Fl
females/group (2 pups per litter)
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
TNO (1998a)
Rat (Wistar); PO, female (28/group)
0, 100, 1,000, 3,000 u.g/L
(equivalent to 0.016, 0.171,
0.489 mg/kg-day, average of
reported intake over premating,
gestation, and lactation)
Drinking water
FO females: 2 weeks prior to mating,
through mating, gestation, and
lactation; FO males: during mating;
Fl animals were not treated after
weaning
Results3
Development of the mammary gland (number of type
day 21 0
day 35 0
day 50 0
day 100 0
1 lobules)
0
30
-1
8
Percent change compared to control
mg/kg-day 0 20
Fl age at vaginal 0 2
opening
100 500
1 3
No significant difference in Fl female sexual maturation as measured by
vaginal opening from PND 32 to 45
Reproductive performance
Gotz et al. (2001)
Rat (Wistar (Crl:WI)),
10-15 pregnant females/group
0, 10 mg/L BBP to pregnant females
during the whole pregnancy and
during lactation
Drinking water
Percent change compared to control
mg/L 0
10
% ofestrus days during
a period of 12 days 12-7 19-7
No change in fertility between groups (quantitative data not reported by
authors).
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Tvl et al. (2004)
Rat (CD); 30 breeding
pairs/group/generation
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
Baver(1998)
Rat (Wistar), 28/sex/group
0, 1, 3 ppm
0, 0.11, 0.35 mg/kg-day for drinking
water
0, 0.09, 0.28 mg/kg-day for diet
Drinking water and diet
Females dosed through mating,
gestation, and lactation (males only
through cohabitation with females)
Nagao et al. (2000)
Rat (Sprague-Dawley);
20-25 breeding pairs/group/
generation; FO reproductive
performance assessed in
25 breeding pairs/group; Fl
reproductive performance assessed
in 20-24 breeding pairs/group
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
Asoetal. (2005)
Rat (Crj:CD(SD)IGS); 24 breeding
pairs/group/generation
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
Results3
Mating or fertility index
mg/kg-day
FO mating index
FO fertility index
Fl mating index
Fl fertility index
Mating or fertility index
(percent change compared to control)
0 50
0 -3
0 7
0 0
0 -3
250
0
4
-4
-7
750
0
4
-28*
-19*
(percent change compared to control)
Drinking water
mg/kg-day
Gestation index
Fertility index
0 0.11
0 0
0 5
0.35
0
14
Diet
mg/kg-day
Gestation index
Fertility index
Mating or fertility index
mg/kg-day
FO mating index (%)
FO fertility index (%)
Fl mating index (%)
Fl fertility index (%)
Mating or fertility index
mg/kg-day
FO mating index
FO fertility index
Fl mating index
Fl fertility index
0 0.9
0 0
0 22
(raw percentages)
0 20
96 96
91.7 83.3
100 94.7
77.3 77.8
100
96
95.8
90.9
95
0.28
0
9
500
100
96
91.7
77.3
(percent change compared to control)
0 100
0 -8
0 4
0 0
0 25
200
-4
9
-9
12
400
-4
10
-9
-15
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Piersma et al. (1995)
Rat (WU); 10 breeding pairs/dose
0, 250, 500, 1,000 mg/kg-day
Gavage
Males: 29 days (14 days premating,
up to 14 days mating); females: up
to 55 days (14 days premating
through PND6)
TNO (1998b)
Rat (Wistar); PO, female (28/group)
0, 1,000, 3,000 u.g/L (equivalent to
0.190, 0.280 mg/kg-day during
premating as calculated by study
authors)
Drinking water
FO females: 2 weeks prior to mating,
through mating, gestation and
lactation; FO males: mating period
only; Fl: did not receive additional
treatment after weaning
TNO (1998a)
Rat (Wistar); PO, female (28/group)
0, 100, 1,000, 3,000 u.g/L
(equivalent to 0.016, 0.171,
0.489 mg/kg-day, average of
reported intake over premating,
gestation, and lactation)
Drinking water
FO females: 2 weeks prior to mating,
through mating, gestation, and
lactation; FO males: during mating;
Fl animals were not treated after
weaning
Results3
Mating or fertility index (raw percentages)
mg/kg-day 0 250 500
FO mating index (%) 100 100 90
FO fertility index (%) 90 80 78
1,000
90
44
Percent change compared to control
mg/kg-day 0 0.190
PO female duration of 0 0
gestation
PO female fecundity 96 82
index (%)
PO female fertility 93 82
index (%)
PO female gestation 100 96
index (%)
PO female mating 96 100
index (%)
PO female pre-coital 0 7
time
0.280
1
96
93
92
96
23
Percent change compared to control
mg/kg-day 0 0.016 0.171
PO female duration of 0 0 1
gestation
Fl female estrus cycle 0 -4 -4
length
PO female pre-coital 0 8 -14
time
0.489
0
-5
-6
Raw percentages
PO female fecundity 96 82 88
index (%)
PO female fertility 89 82 82
index (%)
PO female gestation 96 100 96
index (%)
PO female mating index 93 100 93
(96)
86
86
100
100
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Monsanto (1993)
Rat (Wistar); 12 males/group
0, 0.2, 0.4, 0.8% BBP or
0, 116, 235, 458 mg/kg-day (F) and
0, 252, 580, 1,078 mg/kg-day (M)
Diet
Multigenerational study
Ahmad et al. (2014)
Rat (Albino); PO, female (6/group)
0, 4, 20, 100 mg/kg
Gavage
GD 14 to parturition
Saillenfait et al. (2003)
Rat (Sprague-Dawley); PO, female
(9-10/group)
0, 1.8, 3.6, 5.4 mmol/kg (equivalent
to 560, 1,120, 1,690 mg/kg as
calculated by study authors)
Gavage
Single dose on GD 10; sacrificed
GD21
Saillenfait et al. (2003)
Mouse (OF-1); PO, female
(22-24/group)
0, 0.9, 1.8, 3.6, 5.4 mmol/kg
(equivalent to 280, 560, 1,120,
1,690 mg/kg as calculated by study
authors)
Gavage
Single dose on GD 8; sacrificed
GDIS
Results3
Response
PO females pregnant 25/26 23/28 23/26
24/28
Fertility index (raw percentages)
mg/kg-day 0 116 235
litter 1 92 79 83
litter 2 88 96 88
458
75
92
PO female gestation length was significantly longer in all BBP-treated
groups (graphical presentation reported by study authors)
Percent pregnant (raw percentages)
mg/kg-day 0 560 1,120
PO female percent 100 78 90
pregnant
Note: statistical significance not evaluated by study authors
1,690
100
Percent pregnant (raw percentages)
mg/kg-day 0 280 560 1,120
PO female percent 82 83 70 83
pregnant
Note: Statistical significance not evaluated by study authors
1,690
68
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results3
Biomarkers of reproductive development
Asoetal. (2005)
Rat (Crj:CD(SD)IGS); 24 breeding
pairs/group/generation; AGO
assessed in 19-21 Fl litters/group
and 13-20 F2 litters/group
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
Nagao et al. (2000)
Rat (Sprague-Dawley);
20-25 breeding
pairs/group/generation; AGO
assessed in 128-167 Fl female
pups/group
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
Tvl et al. (2004)
Rat (CD); 30 breeding
pairs/group/generation; AGO
assessed in 26-28 Fl litters/group
and 17-29 F2 litters/group
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
AGO (percent change compared to control)
mg/kg-day 0 100 200
FlAGDatPND4 0 10* 8
Fl AGD/BW1/3 at 0 10* 9*
PND4
F2AGDatPND4 0 -9 -6
F2 AGD/BW1/3 at 0 -5 -9
PND4
400
6
8*
-3
1
AGO (percent change compared to control)
mg/kg-day 0 20 100
500
Fl AGO at birth 0-800*
Note: Fl AGO at birth reported as significantly increased at 500 mg/kg-day
by study authors (pg. 518), but data reported in table do not indicate an
increase.
AGO (percent change compared to control)
mg/kg-day 0 50 250
FlAGDatPNDO 01-4
F2AGDatPNDO 0 -1 -2
750
-4
1
Pregnancy outcomes
Nagao et al. (2000)
Rat (Sprague-Dawley);
20-25 breeding
pairs/group/generation
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
mg/kg-day 0 20 100
Number of implantations/litter (percent change compared to
FO dams for Fllitter 0 6 11
Fl dams for F2 litter 05-4
500
control)
6
-6
Number of live pups/litter (percent change compared to control)
FO dams for Fllitter 0 6 14
Fl dams for F2 litter 04-9
7
-11
Viability during PNDs 0-4 (%) (raw percentages)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Piersma et al. (1995)
Rat (WU); 10 breeding pairs/dose
0, 250, 500, 1,000 mg/kg-day
Gavage
Males: 29 days (14 days premating,
up to 14 days mating); Females: up
to 55 days (14 days premating
through PND6)
Asoetal. (2005)
Rat (Crj:CD(SD)IGS); 24 breeding
pairs/group/generation
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
Saillenfait et al. (2003)
Rat (Sprague-Dawley); PO, female
(9-10/group)
0, 1.8, 3.6, 5.4 mmol/kg (equivalent
to 560, 1,120, 1,690 mg/kg as
calculated by study authors)
Gavage
Results3
FO dams for Fl litter 100
Fl dams for F2 litter 97.8
mg/kg-day 0
Number of implantations/litter
FO dams for Fl litter 0
Fl dams for F2 litter 0
Number of live pups/litter (PND
FO dams for Fl litter 0
Fl dams for F2 litter 0
99 99.5
95.4 99.7
50 250
(percent change compared to
-11 -5
-5 -4
96.7*
97.6
750
control)
-10
-22*
0) (percent change compared to control)
-8 1
-1 0
-2
-20*
4-Ray survival index (%) (raw percentages)
FO dams for Fl litter 97.2
Fl dams for F2 litter 98.3
96.3 97.5
98.1 96.9
92.6
95.4
Percent change compared to control
mg/kg-day 0
number of 0
implan tations/dam
number of live 0
pups/litter
250 500
19 -6
21 -11
1,000
37
-84*
Raw percentages
postnatal mortality 2. 1
PNDs 1-6 (%)
mg/kg-day 0
Number of implantations/litter
FO dams for Fl litter 0
Fl dams for F2 litter 0
2.6 4.8
100 200
(percent change compared to
-13 -6
6 -19
46.7
400
control)
-6
-4
Percent change compared to control
mg/kg-day 0
POfemale 0
implants/litter
560 1,120 1,690 1,690
921
22
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Single dose on GD 10; sacrificed
GD21
Baver(1998)
. . . .
Rat (Wistar), 28/sex/group
0, 1, 3 ppm
0, 0.11, 0.35 mg/kg-day for drinking
water
0, 0.09, 0.28 mg/kg-day for diet
Drinking water and diet
Females dosed through mating,
gestation, and lactation (males only
through cohabitation with females)
Monsanto (1993)
Rat (Wistar); 12 males/group
0, 0.2, 0.4, 0.8% BBP or
0, 116, 235, 458 mg/kg-day (F) and
0, 252, 580, 1,078 mg/kg-day (M)
Diet
Multigenerational study
Saillenfait et al. (2003)
Mouse (OF-1); PO, female
(22-24/group)
0,0.9, 1.8, 3.6, 5.4 mmol/kg
(equivalent to 280, 560, 1,120,
1,690 mg/kg as calculated by study
authors)
Gavage
Single dose on GD 8; sacrificed
GDIS
Ahmad et al. (2014)
Rat (Albino); PO, female (6/group)
0, 4, 20, 100 mg/kg
Gavage
GD 14 to parturition
Results3
Lacation index (percent change compared to control)
Drinking water
mg/kg-day 0 0.11 0.35
00-6
Diet
mg/kg-day 0 0.09 0.28
0 -8 -3
Live birth index (raw percentages)
mg/kg-day 0 116 235 458
litter 1 97 98 100 99
litter 2 98 98 99 99
Viability index (raw percentages)
litter 1 100 97 100 97
litter 2 98 96 100 99
Percent change compared to control
mg/kg-day 0 280 560 1,120 1,690
PO female implants/ 0 -3 -9 -4 9
litter
Percent change compared to control
mg/kg-day 0 4 20 100
Fl combined litter size 0 -1 -5 15
1
2
3
4
*Statistically different from controls (p <0.05), as reported by study authors.
aPercent change compared to control calculated as 100 x ((treated value - control value) 4- control value).
b Calculated as follows: [% in diet x intake food/water (mg)] -f body weight (kg) = mg/kg-day.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 "Values reported by the study authors were estimated from published graphs using "Grab It!", a Microsoft Excel
2 based free software application used to digitizes data from image files. Publisher: www.datatrendsoftware.com.
3
4 Mating index = (number copulated/number cohabitated) x 100; fertility index = (number of pregnant/number
5 copulated) x 100,; gestation index = (number of pregnant females/number of sperm-positive females) x 100;
6 lactation index = (number of live pups after three weeks/number of live pups after four days (after culling)) x 100;
7 fecundity index = (number of females pregnant/number of females mated) x 100; viability index = (number of live
8 pups on day 21/number of live pups on day 4 (after culling)) x 100
9
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
i Ovarian and Uterine weights
Pregnancy Outcomes
BIBRA, 1978; 14wks; gonad re! wt Wistarrats
Ema et al., I998;GD 0-8; ovary & uterine wt Wistar rats
NTP 1989; CD 6-15; gravid uterine wt S-D rats
Aso et al., 2005; 2-gen; FO rel uterus wt S-D rat
Aso et a!., 2005; 2-gen; FO, Fl ovary & uterus abs wt; FO, Fl ovary rel wt; F! uterus
wt S-D rat
Nagaoct at., 2000; 2-gen; FO&FI ovary wt, Fl uterus wt S-D rats
Nagao et at., 2000; 2-gen; FO&FI uterus wt, FO ovaries wt S-D rats
Tyl et
-------
Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
?
Nagao el a),, 2000; 2-gen; Fl ago ; 2-gen; 1 AGO and AGD/BW at PND 4; F2 S-D Rats
Nng.ioet.iI., 2000; 2-gen; AGDatbirtii: T! S-Dr.its
Tylelai., 2004; 2-gen; AC Dal PNDO: K3&F2CD rats
2
3
= statistically significant
-not statistically significant
Figure 3-6. Exposure response array of other female reproductive parameters
following oral exposure to BBP.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
2
3
Table 3-23. Evidence pertaining to pregnancy outcomes following oral
exposure to BBP: Measures of embryotoxicity
Reference and study design
NTP (1989)
Rat (Sprague-Dawley);
27-30 pregnant females/dose
0, 420, 1,100, 1,640 mg/kg-day
Diet
GDs 6-15; dams sacrificed on GD 20
Ema et al. (1990)
Rat (Wistar); 13-17 pregnant
females/dose
0, 0.25, 0.5, 1.0, 2.0%
0, 185, 375, 654, 974 mg/kg-day
Diet
GDs 0-20; dams sacrificed on GD 20
Ema et al. (1992b)
Rat (Wistar); 11 pregnant
females/dose
0 [ad libitum controls], 0 [pair fed
controls], or 974 mg/kg-dayb
Diet
GDs 0-20, 0-11, or 11-20; dams
sacrificed on GD 20
Results3
Raw percentages
mg/kg-day 0 420 1,100
resorptions/litter (%) 3.5 3.8 3.8
litters with 32.1 44.4 43.3
resorptions (%)
1,640
40.2*
86.2*
Percent change compared to control
number of live 052
fetuses/litter
-33*
Raw percentages or ratios
mg/kg-day 0 185 375 654
postimplantation 7.6 9.0 16.4 12.1
loss/litter (%)
total loss/litter (%) 12.7 13.6 25.4 19.8
sex ratio (M: F) 100:108 110:118 67:102 73:87
pre-implantation 5.6 5.2 10.5 8.8
loss/litter (%)
974
100*
100*
NA
13.6
Percent change compared to control
number of live 0 -4 -19* -12
fetuses/litter
Note: All litters were lost at the high dose.
-100
Raw percentages or ratios
mg/kg-day 0 0 974 974
(ad (pair (GDs (GDs
libitum) fed) 0-20) 0-11)
postimplantation 9.2 16.7 100* 100*
loss/litter (%)
litters resorbed (%) 0 0 100* 100*
sex ratio (M:F) 57:78 64:68 NA NA
pre-implantation 6.2 6.4 3.4 7.2
loss/litter (%)
974
(GDs
11-20)
13.4
0
67:67
3.9
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema et al. (1992a)
Rat (Wistar); 11-12 pregnant
females/dose
0 [ad libitum controls], 0 [pair fed
controls], or 974 mg/kg-dayb
Diet
CDs 0-20, 0-7, 7-16, or 16-20;
dams sacrificed at GD 20
Ema et al. (1992c)
Rat (Wistar); 10 pregnant
females/dose
0, 500, 750, 1,000 mg/kg-day
Gavage
GDs 7-15; dams sacrificed at GD 20
Ema et al. (1993)
Rat (Wistar); 10 pregnant
females/dose
0, 600, 750, 1,000 mg/kg-day
Gavage
GDs 7-9, 10-12, or 13-15; dams
sacrificed at GD 20
Results3
Percent change compared to ad libitum control
number of live 0 -2 -100
fetuses/litter
* -100* -1
Raw percentages or ratios
mg/kg-day 0 0 974
(ad (pair (GDs
libitum) fed) 0-20)
postimplantation 9.2 16.7 100*
loss/litter (%)
total loss/litter (%) 14.3 22 100*
litters resorbed (%) 0 0 100*
sex ratio (M: F) 57:78 64:68 NA
pre-implantation 6.2 6.4 3.4
loss/litter (%)
974 974 974
(GDs (GDs (GDs
0-7) 7-16) 16-20)
24.8* 55.8* 11.7
29.6* 59.1* 17.6
0 17 0
54:59 37:36 62:70
6.7 6.7 4.1
Percent change compared to ad libitum control
Number of live 0 -2 -100*
fetuses/litter
-16 -50* -2
Raw percentages or ratios
mg/kg-day 0 500
postimplantation 8.2 14.7
loss/litter (%)
litters resorbed (%) 0 0
sex ratio (M: F) 57:64 62:58
750 1,000
81.7* 100*
30 100*
11:14 NA
Percent change compared to control
number of 0 67 833*
resorptions and dead
fetuses/litter
number of live 0 -1 -79*
fetuses/litter
1,050*
-100
Raw percentages or ratios
mg/kg-day 0 600
750 1,000
Postimplantation loss/litter (%)
exposed GDs 7-9 15.5 14.3
exposed GDs 10-12 13.1 7.8
exposed GDs 13-15 11.7 19.5
52.8* 74.3*
32.1* 88.7*
47.2* 62.3*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema et al. (1995)
Rat (Wistar); 10-12 pregnant
females/dose
0, 750, 1,000, 1,250 mg/kg-day
Gavage
CDs 7-9, 10-12, or 13-15; dams
sacrificed at GD 20
Results3
Sex ratio (M:F)
exposed CDs 7-9 65:60 59:65 36:33
exposed CDs 10-12 54:67 48:81 55:44
exposed CDs 13-15 57:71 55:55 41:32
17:21
13:4*
26:29
Number of resorptions and dead fetuses/litter (percent change compared
to control)
exposed CDs 7-9 0 31 363*
exposed CDs 10-12 0 -35 176*
exposed CDs 13-15 0 65 288*
581*
659*
435*
Number of live fetuses/litter (percent change compared to control)
exposed CDs 7-9 0 -1 -45*
exposed CDs 10-12 0 7 -18
exposed CDs 13-15 0 -14 -43*
-70*
-86*
-57*
Percentage of litters resorbed (percent change compared to control)
exposed CDs 7-9 0 0 10
exposed CDs 10-12 0 0 10
exposed CDs 13-15 000
30
70*
20
Raw percentages or ratios
mg/kg-day 0 750 1,000
1,250
Sex ratio (M:F)
exposed CDs 7-9 68:87 45:44 26:27
exposed CDs 10-12 68:87 64:60 20:12*
exposed CDs 13-15 68:87 51:39* 27:29
NA
NA
NA
Post implantation loss/litter (%)
exposed CDs 7-9 17.5 49.2* 69.6*
exposed CDs 10-12 17.5 30.0* 81.8*
exposed CDs 13-15 17.5 45.9* 68.0*
100*
100*
100*
Number of live fetuses/litter (percent change compared to control)
exposed CDs 7-9 0 -43* -66*
exposed CDs 10-12 0 -20 -79*
exposed CDs 13-15 0 -42* -64*
Note: All litters were resorbed at the high dose.
-100*
-100*
-100*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema et al. (1998)
Rat (Wistar); 7-10 pregnant
females/dose
0, 250, 500, 750, 1,000, mg/kg-day
Gavage
GDs 0-8; dams sacrificed on GD 20
Piersma et al. (2000)
Rat (Harlan Cpb-WU); 4-10 pregnant
females/dose
0, 270, 350, 450, 580, 750, 970,
1,250, 1,600, 2,100 mg/kg-day
GDs 6-15 or 6-20; dams sacrificed
onGD21
Uriu-Adams et al. (2001)
Rat (Wistar); 9-17 pregnant
females/dose
0, 250, 1,000, 1,500,
2,000 mg/kg-day
Gavage
GDs 11-13; dams sacrificed on
GD20
Results3
Raw percentages or ratios
mg/kg-day 0 250
pre-implantation 4.7 5.5
loss/litter (%)
postimplantation 7.2 6.5
loss/litter (%)
sex ratio of live 79:62 72:57
fetuses (M:F)
litters resorbed (%) 0 0
Percent change compared to control
number of live 0 1
fetuses/litter
number of dead or 0 -9
resorbed fetuses/
litter
The study authors reported dose-dependent
resorptions for both exposure periods, with :
dose (data shown graphically).
Resportions (raw percentages)
mg/kg-day 0 250
resorptions (%) 13.08 7.82
Percent change compared to control
number of 0 -9
resorptions/litter
number of live 0 10
fetuses/litter
500 750 1,000
4.0 8.1 16.9*
18.6 29.7* 43.7*
55:54 32:40 18:30*
11 0 0
-14 -27 -51*
164 236 409
increases in numbers of
.00% resorption at the high-
1,000 1,500 2,000
11.52 23.84 53.73*
30 247* 653*
-1 -2 -45*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema and Mivawaki (2002)
Rat (Wistar); 16 pregnant
females/dose
0, 250, 500, 1,000 mg/kg-day
Gavage
GDs 15-17; dams sacrificed on
GD21
Tvl et al. (2004)
Rat (CD); 30 breeding
pairs/dose/generation
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
Howdeshell et al. (2008)
Rat (Sprague-Dawley); 4-9 pregnant
females/dose
0, 100, 300, 600, 900 mg/kg-day
Gavage
GDs 8-18; dams sacrificed on GD 18
Results3
Raw percentages or ratios
mg/kg-day 0 250 500
sex ratio (M: F) 127:107 105:111 111:113
postimplantation 6.4 7.9 7.2
loss/litter (%)
1,000
108:93
15.2
Percent change compared to control
number of live 0 -8 -4
fetuses/litter
number of 0 38 13
resorptions/litter
number of dead 0 -50 -50
fetuses/litter
-14*
138
100
Raw percentages
mg/kg-day 0 50 250
Fl post implantation 15.79 17.65 8.77
loss/litter (%)
F2 post implantation 10.02 8.75 6.67
loss/litter (%)
750
14.18
7.06
Percent change compared to control
Fl number of 0 -11 -5
implan tations/litter
F2 number of 0 -5 -4
implan tations/litter
Fl number of live 0-81
pups/litter
F2 number of live 0 -1 0
pups/litter
-10
-22*
-2
-20*
Raw percentages
mg/kg-day 0 100 300 600
fetal mortality (%) 2.9 0 2.2 12.2*
900
33.3*
Percent change compared to control
number of 099 -15
implan tations/litter
-15
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
TNO (1998b)
Rat (Wistar); PO, female (28/group)
0, 1,000, 3,000 u.g/L (equivalent to
0.190, 0.280 mg/kg-day during
premating as calculated by study
authors)
Drinking water
FO females: 2 weeks prior to mating,
through mating, gestation, and
lactation; FO males: mating period
only; Fl: did not receive additional
treatment after weaning
Results3
number of live 0 11 7 -24* -64*
fetuses
total resorptions 0 -100 0 275* 900*
Response
mg/kg-day 0 0.190 0.280
PO females with all 0 1 2
stillborn pups
PO females with 524
stillborn pups
PO females, stillborn 13 8 28*
PO female, live born 286 240 249
number of litters lost 125
entirely days 0-7
Fl combined sex 154 123 128
ratio (number of
males)
Response (% ± SE)
PO female 10.74 (± 2.823) 11.19 (±4.423) 17.88 (± 5.563)
postimplantation
loss
Response (%)
Fl combined pup 10 4.6* 17*
mortality, day 4 (%)
Fl combined, 100 100 100
Viability index,
days 4-7 (%)
PO female maternal No significant effect on female body weight
body weight throughout the treatment period
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
TNO (1998a)
Rat (Wistar); PO, female (28/group)
0, 100, 1,000, 3,000 u.g/L (equivalent
to 0.016, 0.171, 0.489 mg/kg-day,
average of reported intake over
premating, gestation, and lactation)
Drinking water
FO females: 2 weeks prior to mating,
through mating, gestation, and
lactation; FO males: during mating;
Fl animals were not treated after
weaning
Ahmad et al. (2014)
Rat (Albino); PO, females (6/group)
0, 4, 20, 100 mg/kg
Gavage
GD 14 to parturition
Results3
Response
mg/kg-day
PO females with all
stillborn pups
PO females with
stillborn pups
POfemale, live born
Fl combined pup
mortality, day 4
(number of pups)
Fl combined sex
ratio (number of
males)
POfemale, stillborn
POfemale,
postimplantation
loss
0
1
4
237
2
121
15
16.22
(±4.273)
0.016
0
0
233*
2
125
0*
9.33
(± 1.883)
0.171
0
0
212*
30*
106
0*
13.87
(±4.421)
0.489
0
5
241
29*
126
7
11.34
(±2.755)
Response
mg/kg-day
Fl combined sex
ratio (M/F)
0
0.47
4
0.52
20
0.55
100
0.59
Response (% ± SE)
Fl combined fetal
mortality (%)
Fl combined live
birth index (%) PND 1
Fl combined, live
pups/litter
Fl combined viability
index PND 4
Fl combined
weanling index (%)
PND 21
POfemale, maternal
body weight gain
4
(±4)
96
(±4)
8
(± 1.22)
94
(±6)
94
(±6)
2.78
(±2.78)
97.22
(±2.78)
8.17
(±1.19)
93.17
(±3.17)
80.48
(± 8.01)
Body weight gain was si§
groups
8.21
(±5.64)
91.79
(±5.64)
7.4
(±0.93)
89.79
(±5.25)
81.21
(± 13.4)
;nificantly lower
7.47
(±3.53)
92.53
(±3.53)
8.83
(± 0.4)
89.19
(±3.69)
89.19
(±3.69)
intheBBP
on GD 21 compared to controls
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Saillenfait et al. (2003)
Rat (Sprague-Dawley); PO, female
(9-10/group)
0, 1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 560, 1,120, 1,690 mg/kg as
calculated by study authors)
Gavage
Single dose on GD 10; sacrificed
GD21
NTP (1990)
Mice (CD-I); 27-30 pregnant
females/dose (except n = 14 in the
high-dose group)
0, 182, 910, 2,330, 4,121 mg/kg-day
Diet
GDs 6-15; dams sacrificed on GD 17
Saillenfait et al. (2003)
Mouse (OF-1); PO, female
(22-24/group)
0, 0.9, 1.8, 3.6, 5.4 mmol/kg
(equivalent to 0, 280, 560, 1,120,
1,690 mg/kg as calculated by study
authors)
Gavage
Single dose on GD 8; sacrificed on
GDIS
Results3
Response (% ± SE) or percent change compared to control
mg/kg-day 0 560
PO female percent of 6.06 6.62
postimplantation (± 1.27) (± 1.46) 1
loss/litter
PO female percent of 6.06 6.62
resorptions/litter (± 1.27) (± 1.46) 1
PO female live 0 9
fetuses/litter
1,120
13.86
±0.91)
13.86
±0.91)
-7
1,690
15.55
±3.87)
15.55
±3.87)
-9
Raw percentages
mg/kg-day 0 182
litters with 55 46
resorptions (%)
resorptions/litter (%) 1 4.7
910
63
11.8
2,330
100*
91.3*
Percent change compared to control
number of live 0 4 -9*
fetuses per litter
Note: The 4,121 mg/kg-day group was eliminated after evaluation
14 dams, since all litters were completely resorbed.
-77*
of
Response (% ± SE) or percent change compared to control
mg/kg-day 0 280 560
PO female, percent 5.85 10.89 22.27*
of postimplantation (±2.19) (±2.61) (±5.24)
loss/litter
PO female percent of 5.53 10.02 19.13*
resorptions/litter (±2.05) (±4.56) (±4.56)
PO female live 0 -6 -23
fetuses/litter
1,120
50.41*
(±7.53)
48.1*
(±7.91)
-48*
1,690
77.40*
(±5.14)
73.76*
(±4.78)
-75*
1
2
3
4
5
6
*Statistically different from controls (p <0.05), as reported by study authors.
aPercent change compared to control calculated as 100 x ((treated value - control value) 4- control value).
bCalculated as follows: [% in diet x intake food/water (mg)] -f body weight (kg) = mg/kg-day.
AL=
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
EMBRYOTOXIC3
H
4- Implantations
"0
o
3
"EL
aT
1
o"
t—
o
3
"H-
ET
1
o"
t—
o
CD
O
1
g^s
Howdcshcll ct al, 2008; GD 8-18; S-D rats
Tyl ct al, 2004; 2-gcn; PO/F1 CD rats
Tyl et al., 2004; 2-gen; F1/F2 CD rats
Ema et al, 1990; GD 0-20; Wistar rats
Ema et al, 1 992 b; GD 0-20, GD 0-1 1; Wistar rats
Ema et al, 1 992 b; GD 1 1 -20; Wistar rats
Ema ct al, 1992a; GD 0-20, 0-7, or GD 7-16; Wistar rats
Ema ct al, 1992a; GD 16-20; Wistar rats
Ema et al, 1998; GD 0-8; Wistar rats
Ema ct al, 1 992c; GD 7-15; Wistar rats
Ema et al, 1993; GD 7-9, GD 10-12, or GD 13-15; Wistar rats
Ema ctal, 1995;GD 7-9, GD 10-12, orGD13-15; Wistar rats
Saillenfait, 2003; GD 10; S-D rats
TNO, 1998a; 2-gcn; PO Wistarrats
TNO, 1998b; 2-gcn; PO Wistar rats
Tyl et al., 2004; 2-gcn; PO/F1 and F1/F2 CD rats
Saillenfait, 2003; GD 8; OF-1 mice
Ema et al, 1990; GD 0-20; Wistar rats
Ema ct al, 1 992 b; GD 0-20, GD 0-1 1, or GD 1 1 -20; Wistar rats
Ema ct al, 1992a; GD 0-20, 0-7, GD 7-16, or GD 16-20; Wistar rats
Ema et al, 1 998; GD 0-8; Wistar rats
Ema ct al, 1992c; GD 7-15; Wistar rats
Ema et al, 1998; GD 0-8; Wistar rats
Howdcshcll et al, 2008; GD 8-18; S-D rats
NTP 1 989; GD 6-15; S-D rats
Ema ctal, 1992b;GDO-ll orGD 0-20; Wistarrats
Uriu-Adamset al, 2001 ; GD 1 1 -13; Wistar rats
NTP, 1990; GD 6-15; CD-I mice
Saillenfait, 2003; GD 8; OF-1 mice
Ema et al, 1990; GD 0-20; Wistar rats
Bn n
Q
Q
a a am
m
a
m
a
B-Bmm
EM
M
«
B-BQ
Qn n
BE)
Qn n
B-B-BO
D
B
D D J
a am
B — B-mm
a — &•
"
B-m-mm
a a am
- statistically significant
= not statistically significant
0.01 0.1 1 10 100 1000 10000
Dose (mg/kg-day)
2
3
Figure 3-7. Exposure-response array of pregnancy outcomes following oral
exposure to BBP.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
2
3
Etna ct al, 1990; GD 0-20; Wistar rats
Ema ct al., 1992a; GD 0-20 or GD 7-16; Wistar rats
Ema ct al., 1992a; GD 0-7 orGD 16-20; Wistar rats
llowdcshcll ct al., 2008; GD 8-18; S-D rats
Saillcnfait, 2003; GD 10; S-D rats
Uriu-Adams ct al., 2001; GD 11-13; Wistar rats
Ema and Miyawaki, 2002; GD 15-17; Wistar rats
TNO, 1998a; 2-gcn; PO Wistar rats
TNO, 1998b; 2-gen; PO Wistar rats
NTP, 1990; GD 6-15; CD-I mice
Saillcnfait, 2003; GD 8; OF-1 mice
m
1 I
i!
ix ^
5 2
3 S
K.
TNO, 1998a; 2-gcn; Fl Wistar rats
TNO, 1998b; 2-gcn; Fl Wistar rats
Ilowdcshcll ct al., 2008; GD 8-18; S-D rats
Ahmad, 2014; GD 14-parturition; albino rats
Ema ct al., 1990; GD 0-20; Wistar rats
Ema ct al., 1993; GD 7-9 or GD 13-15; Wistar rats
Ema ct al., 1992c; GD 7-15; Wistar rats
Ema ct al, 1992 b; GD 11 -20; Wistar rats
Ema ctal., 1993; GD 10-12; Wistar rats
Ema ct al., 1995; GD 7-9; Wistar rats
Ema ct al., 1995; GD 10-12; Wistar rats
Ema ctal., 1995; GD 13-15; Wistar rats
Ema and Miyawaki, 2002; GD 15-17; Wistar rats
Ahmad, 2014; GD 14-parturition; albino rats
Q B-
Q—B-B
-B O
Da:
B—B—B
= statistically significant 0.01
-not statistically significant
0.1 1 10 100
Dose (mg/kg-day)
1000 10000
Figure 3-8. Exposure-response array of fetal measures following oral
exposure to BBP.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.3.3. Developmental Effects
2
3
Table 3-24. Evidence pertaining to developmental effects following oral
exposure to BBP: Teratogenicity
Reference and study design
NTP (1989)
Rat (Sprague-Dawley);
27-30 pregnant females/dose
0, 420, 1,100,1,640 mg/kg-day
Diet
GDs 6-15; dams sacrificed on GD 20
Ema et al. (1990)
Rat (Wistar); 13-17 pregnant
females/dose
0, 0.25, 0.5, 1.0, 2.0%
0, 185, 375, 654, 974 mg/kg-day
GDs 0-20; dams sacrificed on GD 20
Results3
Raw percentages
mg/kg-day 0 420
fetuses 2.0 0.9
m alform ed /litter
litters with 25.0 14.8
malformed fetuses
fetuses with 19.0 25.4
variations/litter (%)
1,100 1,640
5.9 52.8*
46.7 96.3*
41.0* 71.4*
Percent incidence
number of litters 0 0
with external
malformation
number of litters 11 11
with skeletal
malformations
number of litters 18 4
with visceral
malformations
10 52*
30 89*
33 78*
Percent incidence
mg/kg-day 0 185
number of litters 0 6
with external
anomalies
number of litters 13 6
with skeletal
anomalies
number of litters 40 47
with skeletal
variation
number of litters 20 35
with delayed
ossification in
sternebrae
375 654
0 15
13 23
27 69
27 38
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema et al. (1992b)
Rat (Wistar); 11 pregnant
females/dose; 132-135, 88-89, and
44-46 fetuses/group examined for
external, skeletal, and internal
malformations, respectively
0 ad libitum controls, 0 pair fed
controls, or 974 mg/kg-dayb
Diet
CDs 0-20, 0-11, or 11-20; dams
sacrificed on GD 20
Ema et al. (1992a)
Rat (Wistar); 11-12 pregnant
females/dose; 73-135, 49-90, and
24-46 fetuses/group examined for
external, skeletal, and internal
malformations, respectively
0 ad libitum controls, 0 pair fed
controls, or 974 mg/kg-dayb
Diet
Results3
number of litters 0
with internal
anomalies
Note: All litters were lost at the
0 0
high dose.
8
Percent litter Incidence
mg/kg-day 0
(ad
libitum)
external 9
malformations
cleft palate 0
skeletal 0
malformations
fused sternebrae 0
internal 0
malformations
0 974 974
(pair fed) (GDs (GDs
0-20) 0-11)
0 NA NA
0 NA NA
18 NA NA
9 NA NA
0 NA NA
974
(GDs
11-20)
82*
82*
82*
73*
0
Percent fetal incidence
mg/kg-day 0
(ad
libitum)
all external 1
malformations
cleft palate 0
all skeletal 0
malformations
fused sternebrae 0
internal 0
malformations
0 974 974
(pair (GDs (GDs
fed) 0-20) 0-11)
1 NA NA
0 NA NA
2 NA NA
1 NA NA
0 NA NA
974
(GDs
11-20)
54*
54*
27*
25*
0
Note: All litters were lost in groups treated from GDs 0-20 and 0-11.
Percent litter incidence
mg/kg-day 0
(ad
libitum)
external 9
malformations
cleft palate 0
skeletal 0
malformations
0 974 974 974
(pair (GDs (GDs (GDs
fed) 0-20) 0-7) 7-16)
0 NA 0 100*
0 NA 0 100*
18 NA 9 90*
9974
(GDs
16-20)
0
0
18
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
CDs 0-20, 0-7, 7-16, or 16-20;
dams sacrificed on GD 20
Ema et al. (1992c)
Rat (Wistar); 10 pregnant
females/dose; 25-121, 16-81, and
9-41 fetuses/group examined for
external, skeletal, and internal
malformations, respectively
0, 500, 750, 1,000 mg/kg-day
Gavage
GDs 7-15; dams sacrificed on GD 20
Results3
fused sternebrae 0
internal 0
malformations
9 NA 0 90*
0 NA 0 22
0
0
Percent fetal incidence
external 1
malformations
cleft palate 0
skeletal 0
malformations
fused sternebrae 0
internal 0
malformations
Note: All litters were lost in the
0 NA 0 93*
0 NA 0 93*
2 NA 1 78*
1 NA 0 78*
0 NA 0 8
group treated from GDs 0-20.
0
0
3
0
0
Percent litter incidence
mg/kg-day 0
external 0
malformations
cleft palate 0
skeletal 10
malformations
fused sternebrae 0
internal 0
malformations
dilation of renal 0
pelvis
500 750
0 100*
0 100*
30 57*
0 57*
0 60*
0 60*
1,000
NA
NA
NA
NA
NA
NA
Percent fetal incidence
external 0
malformations
cleft palate 0
skeletal 1
malformations
fused sternebrae 0
internal 0
malformations
dilation of renal 0
pelvis
0 48*
0 48*
4 31*
0 25*
0 33*
0 33*
NA
NA
NA
NA
NA
NA
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema et al. (1993)
Rat (Wistar); 10 pregnant
females/dose; 38-125, 25-83, and
13-42 fetuses/group examined for
external, skeletal, and internal
malformations, respectively
0, 600, 750, 1,000 mg/kg-day
Gavage
CDs 7-9, 10-12, or 13-15; dams
sacrificed on GD 20
Results3
Note: All litters were lost at the high dose.
Percent litter incidence
mg/kg-day 0
600
750
1,000
External malformations
CDs 7-9 0
CDs 10-12 0
CDs 13-15 0
0
0
10
0
11
70*
29
33
100*
Skeletal malformations
CDs 7-9 10
CDs 10-12 10
CDs 13-15 0
30
20
20
56*
11
70*
86*
0
100*
Internal malformations
CDs 7-9 0
CDs 10-12 0
CDs 13-15 0
0
0
0
11
11
0
29
0
0
Percent fetal incidence
External malformations
CDs 7-9 0
CDs 10-12 0
CDs 13-15 0
0
0
2
0
1
47
5
6
82
Skeletal malformations
CDs 7-9 1
CDs 10-12 1
CDs 13-15 0
5
2
5
20
2
42
44
0
97
Internal malformations
CDs 7-9 0
CDs 10-12 0
CDs 13-15 0
0
0
0
4
3
0
Note: Specific malformations that were significantly increased
vertebral malformations (GDs 7-9), fusion of sternebrae (GDs
cleft palate (GDs 13-15).
15
0
0
included
13-15), and
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema et al. (1995)
Rat (Wistar); 10-12 pregnant
females/dose; 53-155, 35-102, and
18-53 fetuses/group examined for
external, skeletal, and internal
malformations, respectively
0, 750, 1,000, or 1,250 mg/kg-day
Gavage
CDs 7-9, 10-12, 13-15; dams
sacrificed on GD 20
Results3
Percent litter incidence
mg/kg-day 0
750
1,000
1,250
External malformations
CDs 7-9 0
CDs 10-12 0
CDs 13-15 0
0
9
67*
33
40
100*
NA
NA
NA
Skeletal malformations
CDs 7-9 8
CDs 10-12 8
CDs 13-15 8
64*
9
67*
89*
0
100*
NA
NA
NA
Internal malformations
CDs 7-9 0
CDs 10-12 0
CDs 13-15 0
18
9
0
22
0
0
NA
NA
NA
Percent fetal incidence
External malformations
CDs 7-9 0
CDs 10-12 0
CDs 13-15 0
0
1
48
6
6
82
NA
NA
NA
Skeletal malformations
CDs 7-9 1
CDs 10-12 1
CDs 13-15 1
19
1
44
54
0
97
NA
NA
NA
Internal malformations
CDs 7-9 0
CDs 10-12 0
CDs 13-15 0
Note: All litters were resorbed at
that were significantly increased
7
2
0
11
0
0
NA
NA
NA
the high dose. Specific malformations
included vertebral
(GDs 7-9), fusion/absence of ribs (GDs 7-9), fusion
malformations
of sternebrae
(CDs 13-15), and cleft palate (GDs 13-15).
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Piersma et al. (2000)
Rat (Harlan Cpb-WU); 4-10 pregnant
females/dose
0, 270, 350, 450, 580, 750, 970,
1,250, 1,600, 2,100 mg/kg-day
GDs 6-15 or 6-20; dams sacrificed
onGD21
Uriu-Adams et al. (2001)
Rat (Wistar); 9-17 pregnant
females/dose; 8-16 litters/dose
(36-119 fetuses/dose) were
examined for anomalies and
malformations
0, 250, 1,000, 1,500,
2,000 mg/kg-day
Gavage
GDs 11-13; dams sacrificed on
GD20
Saillenfait et al. (2003)
Rat (Sprague-Dawley); Fl, combined
(100-123/group)
0, 1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 560, 1,120, 1,690 mg/kg as
calculated by study authors)
Results3
The study authors reported increased incidences of several skeletal
anomalies in the "middle or high doses" (quantitative data not provided).
There was a dose-dependent increase in the occurrence of extra 13th
lumbar ribs in groups exposed to 270-1,250 mg/kg-day (all litters were
resorbed at higher doses); the effect was more pronounced in the group
exposed on GDs 6-20 (data shown graphically).
Percent change compared to control
mg/kg-day 0 250 100 1,500
crown rump length, 0 11-1
males
crown rump length, 001-2
females
2,000
-6*
-10*
Skeletal malformations
ossification sites, 0 -5 -4 -9
metacarpals
ossification sites, 0 -4 -2 -16*
metatarsals
ossification sites, 0 -11 -8 -36*
sternum
number of 0 0 2,150* 6,800*
rudimentary
ribs/fetus
-32*
-27*
-62*
7,250*
Raw percentages
fetuses with rib 1.79 2.38 23.30* 82.64*
anomaly (%)
92.26*
External malformations
fetuses with cleft 0 0 2.3 27.5* 52.9*
palate/litter (%)
Note: Study authors did not report whether the litter or the fetus was the
statistical unit of comparison.
Response (%)
mg/kg-day 0 560 1,120
Fl combined 0 0* 0.9*
percent of
malformed fetuses
1,690
5*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Gavage
Single dose on GD 10; sacrificed
GD21
Saillenfait et al. (2003)
Mouse (OF-1); Fl, combined
(35-221/group)
0, 0.9, 1.8, 3.6, 5.4 mmol/kg
(equivalent to 0, 280, 560, 1,120,
and 1,690 mg/kg as calculated by
study authors)
Gavage
Single dose on GD 8; sacrificed
GDIS
NTP (1990)
Mouse (CD-I); 27-30 pregnant
females/dose (except n = 14 in the
high-dose group)
0, 182, 910, 2,330, 4,121 mg/kg-day
Diet
GDs 6-15; dams sacrificed on GD 17
Results3
Response (%)
mg/kg-day 0 280 560 1,120 1,690
Fl combined, 0 0* 2.1* 9.1* 42.9*
percent of
malformed fetuses
mg/kg-day 0 182 910 2,330
Raw percentages
litters with gross 10 0 27 67*
malformations (%)
litters with skeletal 21 11 43 100*
malformations (%)
litters with visceral 7 7 23 33
malformations (%)
malformed 4.4 2.4 13.6* 89.3*
fetuses/litter (%)
litters with 31 18 60* 100*
malformed fetuses
fetuses with 29 26.2 35.9 98.4*
variations per litter
litters with 86 82 97 100
variations (%)
Note: The 4,121 mg/kg-day group was eliminated after evaluation of
14 dams, since all litters were completely resorbed.
1
2
3
4
5
*Statistically different from controls (p <0.05), as reported by study authors.
aPercent change compared to control calculated as 100 x ((treated value - control value) 4- control value).
bCalculated as follows: [% in diet x intake food/water (mg)] -f body weight (kg) = mg/kg-day.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Ema et al, 1992c; CD 7-15; Wistar rats
NTP 1989; GD6-15;S-D rats
Ema et al, 1990; CDO-20; Wistar rats*
Ema et al, 1992b; CD 11-20; Wistar rats
Ema et al, 1992a; CD 0-7, or GD 16-20; Wistar rats
Ema et al, 1992a; GD7-16; Wistar rats
Ema et al, 1993; GD 13-15; Wistar rats
Ema et al, 1993; GD 7-9 or GD 10-12; Wistar rats
Ema et al, 1995; GD 7-9 or GD 10-12; Wistar rats**
Ema etal, 1995; GDI 3-15; Wistar rats**
Saillenfait, 2003; GD 10; S-D rats
Saillenfait, 2003; GD8; OF-1 mice
NTP, 1990; GD 6-15; CD-I mice
Ema et al, 1992c; GD 7-15; Wistar rats
NTP, 1989; GD 6-1 5; S-D rats
Ema et al, 1990; GDO-20; Wistar rats*
Ema etal, 1992b; GDI 1-20; Wistar rats
Ema etal, 1992a;GD 0-7, or GD 16-20; Wistar rats
Eraa et al, 1992a; GD7-16; Wistar rats
Ema et al, 1993; GD 7-9 or GD 13-15; Wistar rats
Ema etal, 1993; GD 10-12; Wistar rats
Ema etal, 1995; GD 10-12; Wistar rats
Ema et al, 1995; GD 7-9or GD 13-15; Wistar rats
Uriu-Adams etal., 2001; GD 11-13; Wistar rats***
NTP, 1990; GD 6-1 5; CD-I mice
100
•Ema et al 1990- all litters lost at high dose 974 mg/kg-d
** Ema et al 1995 - all litters lost at high dose 1250 mg/kg-d
***Uriu-Adams reported endpoints on a per-fetus basis
""NTP 1990 - all litters lost in 4121 mg/kg-d dose group
1000
Dose (mg/kg-day)
10000
• = statistically significant
:j = not statistically significant
2
3
Figure 3-9. Exposure-response array of developmental effects following oral
exposure to BBP: teratogenicity.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
T Visceral Malformat
o'
E
ariations/
3
O
3
n'
tu
cd
Ema et al., 1992c; GD 7-15; Wistar rats
NTP 1989; GD 6-15; S-D rats
Ema etal., 1992b; GDI 1-20; Wistar rats
Ema et al., 1992a; GDO-7, orGD 16-20; Wistar rats
Ema et al., 1992a; GD7-16; Wistar rats
Ema et al., 1993; GD 7-9, GD 10-12, or GD 13-15;
Wistar rats
Emaetal., 1995; GD 7-9, GD 10-1 2, or GD 13-15;
Wistar rats
NTP, 1990; GD 6-1 5; CD-I mice
Bn ™
D
QPI PI
D
D
ODD
Q— H
*Ema etal 1990-all litters lost at high dose 974 mg/kg-d
** Ema et al 1995 - all litters lost at high dose 1250 mg/kg-d
***Uriu-Adams reported endpoints on a per-fetus basis
**"NTP 1990- all litters lost in 4121 mg/kg-d dose group
100
1000
Dose (mg/kg-day)
10000
• = statistically significant
n - not statistically significant
3
4
Figure 3-10. Exposure-response array of developmental effects following oral
exposure to BBP: malformations.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
2
Table 3-25. Evidence pertaining to developmental effects following oral
exposure to BBP: offspring body weight
Reference and study design
Results3
Fetal body weight
NTP (1989)
Rat (Sprague-Dawley CD);
27-30 pregnant females/dose
0, 420, 1,100, 1,640 mg/kg-day
Diet
GDs 6-15; dams sacrificed on GD 20
Ema et al. (1990)
Rat (Sprague-Dawley); 13-17
pregnant females/dose
0, 0.25, 0.5, 1.0, 2.0%
0, 185, 375, 654, 974 mg/kg-day
Diet
GDs 0-20; dams sacrificed on GD 20
Ema et al. (1992b)
Rat (Wistar); 11 pregnant
females/dose
0 ad libitum controls, 0 pair fed
controls, or 974 mg/kg-day
Diet
GDs 0-20, 0-11, or 11-20; dams
sacrificed on GD 20
Percent change compared to control
mg/kg-day 0 420 1,100
1,640
Fetal body weight (litter means)
all 0 -3 -3
males 0 -3 -3
females 00-3
-21*
-20*
-21*
Percent change compared to control
mg/kg-day 0 185 375 654
974
Fetal body weight (litter means)
males 025* -7*
females 023* -7*
Note: All litters were lost at the high dose.
NA
NA
Percent change compared to control
mg/kg-day 0 0 974 974
(ad (pair (GDs (GDs
libitum) fed) 0-20) 0-11)
974
(GDs
11-20)
Fetal body weight (litter means)
males 0 -8* NA NA -22*
females 0 -10* NA NA -19*
Note: Statistical results are shown for comparison to ad libitum control
group. Fetal body weights in the group treated on GDs 11-20 were also
significantly lower than pair fed controls. All litters were lost in groups
treated on GDs 0-20 and GD 0-11.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema et al. (1992a)
Rat (Wistar); 11-12 pregnant
females/dose
0 ad libitum controls, 0 pair fed
controls, or 974 mg/kg-day
Diet
CDs 0-20, 0-7, 7-16, or 16-20;
dams sacrificed on GD 20
Ema et al. (1992c)
Rat (Wistar); 10 pregnant
females/dose
0, 500, 750, 1,000 mg/kg-day
Gavage
GDs 7-15; dams sacrificed on GD 20
Ema et al. (1993)
Rat (Wistar); 10 pregnant
females/dose
0, 600, 750, 1,000 mg/kg-day
Gavage
GDs 7-9, 10-12, or 13-15; dams
sacrificed on GD 20
Results3
Percent change compared to control
mg/kg-day 0 0 974
(ad (pair (GDs
libitum) fed) 0-20)
974
(GDs
0-7)
974 974
(GDs (GDs
7-16) 16-20)
Fetal body weight (litter means)
males 0 -8* NA -7* -11* -17*
females 0 -10* NA -9* -12* -18*
Note: Statistical results are shown for comparison to ad libitum control
group. Fetal body weights in the groups treated on GDs 16-20 were also
significantly lower than pair fed controls. All litters were lost in the group
treated on GDs 0-20.
Percent change compared to control
mg/kg-day 0 500
750
1,000
Fetal body weight (litter means)
males 0 -5
females 0 -4
Note: All litters were lost at the high dose.
-18*
-18*
NA
NA
Percent change compared to control
mg/kg-day 0 600
750
1,000
Male fetal body weight (litter means)
exposed GDs 7-9 0 -3
exposed GDs 10-12 0 5
exposed GDs 13-15 0 3
-15*
-6
-2
-18*
-14*
-5
Female fetal body weight (litter means)
exposed GDs 7-9 0 -3
exposed GDs 10-12 0 4
exposed GDs 13-15 0 -1
-16*
-4
-5
-16*
-9
-6
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema et al. (1995)
Rat (Wistar); 10-12 pregnant
females/dose
0, 750, 1,000, 1,250 mg/kg-day
Gavage
CDs 7-9, 10-12, or 13-15; dams
sacrificed on GD 20
Ema et al. (1998)
Rat (Wistar); 7-10 pregnant
females/dose
0, 250, 500, 750, 1,000 mg/kg-day
Gavage
GDs 0-8; dams sacrificed on GD 20
Piersma et al. (2000)
Rat (Harlan Cpb-WU);
4-10 pregnant females/dose
0, 270, 350, 450, 580, 750, 970,
1,250, 1,600, 2,100 mg/kg-day
Gavage
GD 6-15 or GD 6-20 ; dams
sacrificed on GD 21
Uriu-Adams et al. (2001)
Rat (Wistar); 9-17 pregnant
females/dose
0, 250, 1,000, 1,500, 2,000
mg/kg-day
Gavage
Results3
Percent change compared to control
mg/kg-day 0 750 1,000
1,250
Male fetal body weight (litter means)
exposed GDs 7-9 0 -14* -17*
exposed GDs 10-12 0 -5 -14*
exposed GDs 13-15 0 -3 -8
NA
NA
NA
Female fetal body weight (litter means)
exposed GDs 7-9 0 -16* -17*
exposed GDs 10-12 0 -5 -15*
exposed GDs 13-15 0 -3 -5
Note: All litters were lost at the high dose.
NA
NA
NA
Percent change compared to control
mg/kg-day 0 250 500 750
1,000
Fetal body weight (litter means)
males 0 0 -14* -32*
females 0 -2 -14* -33*
-45*
-40*
Percent change compared to control
mg/kg- 0 270 350 450 580 750
day
fetal 0 -4 -5 -5 -7 -15
weight13
Note: The study authors reported a dose-dependent decrease
weight for both exposure periods. All litters were resorbed in
and 2,100 mg/kg-day groups.
970 1250
-22 -28
in fetal
the 1,600
Percent change compared to control
mg/kg-day 0 250 1,000 1,500
2,000
Fetal body weight (litter means)
males 02 4-7*
-18*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
GDs 11-13; dams sacrificed on
GD20
Ema and Mivawaki (2002)
Rat (Wistar); 16 pregnant
females/dose
0, 250, 500, 1,000 mg/kg-day
Gavage
GDs 15-17; dams sacrificed on
GD21
NTP (1990)
Mouse (CD-I); 27-30 pregnant
females/dose (except n = 14 in the
high-dose group)
0, 182, 910, 2,330, 4,121 mg/kg-day
Diet
GDs 6-15; dams sacrificed on GD 17
Saillenfait et al. (2003)
Mouse (OF-1); Fl, combined
(35-221/group)
0, 0.9, 1.8, 3.6, 5.4 mmol/kg
(equivalent to 0, 280, 560, 1,120,
1,690 mg/kg as calculated by study
authors)
Gavage
Single dose on GD 8; sacrificed
GDIS
Results3
females
01 3-9*
-22*
Percent change compared to control
mg/kg-day
Fetal body weight (litter
males
females
0 250 500
means)
040
04-1
1,000
-17*
-14*
Percent change compared to control
mg/kg-day
Fetal body weight (litter
0 182 910
means)
All 01-4
males 02-3
females 02-3
Note: The 4,121 mg/kg-day group was eliminated after evaluation
14 dams, since all litters were completely resorbed.
2,330
-17*
-16*
-14*
of
Percent change compared to control
mg/kg-day
Fl combined, mean
fetal weight/litter
0 280 560 1,120
0 -2 -2 -8
1,690
16*
Pup body weight
Piersma et al. (1995)
Rat (WU); 10 breeding pairs/dose
0, 250, 500, 1,000 mg/kg-day
Gavage
Males: 29 days (14 days premating,
up to 14 days mating); females: up
Percent change compared to control
mg/kg-day
mean pup weight on
PND1
mean pup weight on
PND6
0 250 500
0 -1 -7*
03-1
1,000
-29*
-43*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
to 55 days (14 days premating
through PND6)
Bayer (1998)
Rat (Wistar), 28/sex/group
0, 1, 3 ppm
0, 0.11, 0.35 mg/kg-day for drinking
water
0, 0.09, 0.28 mg/kg-day for diet
Drinking water and diet
Females dosed through mating,
gestation, and lactation (males only
through cohabitation with females)
Nagao et al. (2000)
Rat (Sprague-Dawley);
20-25 breeding pairs/group/
generation
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
Tvl et al. (2004)
Rat (CD); 30 breeding
pairs/group/generation
Results3
Note: The statistical unit of comparison (litter
reported.
or individual pup) was not
Percent change compared to control
Drinking water
mg/kg-day 0 0.11
0.35
mean pup weight on PND 0
Males 0 0
Females 0 0
mean pup weight on PND 21
Males 0 2
Females 0 3
3
4
5
6
Diet
mg/kg-day 0 0.09
mean pup weight on PND 0
Males 0 2
Females 0 2
mean pup weight on PND 21
Males 0 -3
Females 0 -3
0.28
3
4
0.4
-0.7
Percent change compared to control
mg/kg-day 0 20
100 500
Fl mean pup weight
males PND 0 0 0 -6* -7*
females PND 0 0 2 -6* -6*
males PND 14 00-1 -8*
females PND 14 01-3 -8*
males PND 21 01-1 -7*
females PND 21 01-2 -7*
Note: There were no significant differences in Fl body weight at PND 4 or
PND 7; there were no significant differences in F2 body weight at any time
point. The study authors did not state whether the litter or the individual
was the statistical unit of comparison.
Percent change compared to control
mg/kg-day 0 50
250 750
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
Asoetal. (2005)
Rat (Crj:CD(SD)IGS); 24 breeding
pairs/group/generation
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
Ahmad et al. (2014)
Rat (Albino); PO, female (6/group)
0, 4, 20, 100 mg/kg
Gavage
GD 14 to parturition
TNO (1998a)
Rat (Wistar); PO, female (28/group)
0, 100, 1,000, 3,000 u.g/L
(equivalent to 0.016, 0.171, 0.489
mg/kg-day, average of reported
intake over premating, gestation,
and lactation)
Drinking water
FO females: 2 weeks prior to mating,
through mating, gestation, and
lactation; FO males: during mating;
Fl animals were not treated after
weaning
TNO (1998b)
Rat (Wistar); PO, female (28/group)
0, 1,000, 3,000 u.g/L (equivalent to
0.190, 0.280 mg/kg-day during
premating as calculated by study
authors)
Results3
Litter mean pup body weight at PND 0
Flmale 03-1 -9*
Fl female 05 1-7*
f 2 male 01 -2-5
F2 female 020-5
Note: Study authors report significant lowered bodyweights on PND 0 in Fl
males >100 mg/kg-day and F2 males and females at 100 and
400 mg/kg-day. (Data shown graphically - Figure 1-4).
Percent change compared to control
mg/kg-day 0 4 20 100
Flmale, pup weight 0 -3* -4* -5*
(M) PND 1
Flmale, pup weight 0 -13* -22* -16*
(M) PND 21
Percent change compared to control
mg/kg-day 0 0.016 0.171 0.489
Fl combined pup 0032
weight PND 1
Fl combined pup 0-110
weight PND 14
Fl female, pup weight 0-51-3
PND 21
Fl male, pup weight 0-130
PND 21
Fl combined pup 0052
weight preculling
Fl combined pup 0-141
weight PND 7
Percent change compared to control
mg/kg-day 0 0.190 0.280
Fl combined, pup 028
weight PND 1
029
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results3
Drinking water
FO females: 2 weeks prior to mating,
through mating, gestation and
lactation; FO males: mating period
only; Fl: did not receive additional
treatment after weaning
Fl combined, pup
weight PND 4
preculling
Fl combined, pup
weight PND 7
1
2
3
4
5
6
*Statistically different from controls (p <0.05), as reported by study authors.
aPercent change from controls calculated as 100 x ((treated value - control value) 4- control value).
bValues reported by the study authors were estimated from published graphs using "Grab It!", a Microsoft Excel
based free software application used to digitizes data from image files. Publisher: www.datatrendsoftware.com.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Ema ct al., 1990; GD 0-20; Wistar rats*
Emactal., 1992c;GD 7-15; Wistar rats**
Ema ct al., 1993; GD 7-9; Wistar rats
Emactal., 1993; GD 10-12; Wistar rats
Ema ct al., 1993; GD 13-15; Wistar rats
Ema ct al., 1998; GD 0-8; Wistar rats
Emactal., 1992b;GD 11-20; Wistar rats
| Ema ct al., 1992a; GD 0-7, GD 7-16, or GD 16-20,
to Wistar rats
o
CL
Ema ct al, 1995; GD 7-9; Wistar rats***
Ema ct al., 1995; GD 10-12; Wistar rats***
Ema ct al., 1995; GD 13-15; Wistar rats***
Ema and Miyawaki, 2002; GD 15-17; Wistar rats
Uriu-Adamsctal., 2001; GD 11-13; Wistar rats
NTP 1989; GD 6-15; S-D rats
Saillcnfait, 2003; GD 8; OF-1 mice
NTP, 1990; GD 6-15; CD-I mice
*Ema et al 1990 - all litters lost at high dose 974 mg/kg-d;
fetal weight significantly increased at 375 but significantly
lower at 654 mg/kg-d
** Ema et al 1992c - all litters lost at high dose
***Ema et al 1995 - all litters lost at high dose 1250 mg/kg-d
• = statistically significant
n = not statistically significant
B-B
QBQ
Q B-
Q B-
B B B-
10 100 1000
Dose (mg/kg-day)
10000
2
3
Figure 3-11. Exposure-response array of developmental effects following oral
exposure to BBP: fetal body weight.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
"d
03
o
D.
tro'
Piersma ct al., 1 995; premating-PND6; pup bw PND
l;WUrats
Piersma ct al., 1 995; premating-PND6; pup bw PND
6; WU rats
Nagao ct al., 2000; 2-gen; Fl pup bw PND 0; S-D rats
Nagao et al., 2000; 2-gen; Fl pup bw PND 14&21 S-
D rats
Tyl ct al., 2004; 2-gen; Fl pup bw PND 0; CD rats
Tylctal., 2004; 2-gen; F2 pup bw PND 0; CD rats
Ahmad, 201 4; GDI 4-parturition; pup bw PND1 &21
albino rats
TNO, 1998a; 2-gen; Fl pup bw PND 1, 4. 7; Wistar
rats
TNO, 1998b; 2-gen; Fl pup bw PND 1, 7, 14, 21;
Wistar rats
Aso ct al., 2005; 2-gcn; Fl male pup bw PND 0; S- D
rats
Aso ct al., 2005; 2-gen; F2 pup bw PND 0; S-D rats
B-M
D-B-B
GH-*-.
G B •
Q B— •
Q B-G
• • •
ED
—
•Emaetal 1990 -all litters lost at high dose 974 mg/kg-d; Q QJ QJ ^ 1
fetal weight significantly increased at 375 but significantly
lower at 654 mg/kg-d
** Ema et al 1992c - all litters lost at high dose Dose fme/kfi-davl
***Emaetal 1995 -all litters lost at high dose 1250 mg/kg-d
• = statistically significant
^ - not statistically significant
IQQ 1Q0010000
2
3
Figure 3-12. Exposure-response array of developmental effects following oral
exposure to BBP: pup weight.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
3.3.4. Liver Effects
2
3
Table 3-26. Evidence pertaining to liver effects in animals following oral and
inhalation exposure to BBP
Reference and study design
Results3
Liver weightb
NTP (1989)
Rat (Sprague-Dawley CD);
30 females/group
0, 420, 1,100, 1,640 mg/kg-day
Diet
CDs 6-15
BIBRA (1978)
Rat (Wistar); 27/sex/group or
45/sex/group (control); interim
sacrifices of
9 controls/sex/group and
6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000 12,000 ppm
0, 151, 381, 960 mg/kg-day
(males)
0, 171, 422, 1,069 mg/kg-day
(females)
Diet
14 weeks
Maternal liver weight,
mg/kg-day
absolute weight
relative weight
GD
0
0
0
20 (percent change compared to control)
420
4
0
1,100
5
7*
1,640
1
13*
Liver weight (percent change compared to control)
mg/kg-day (M)
absolute weight;
2 weeks
absolute weight;
6 weeks
absolute weight;
14 weeks
relative weight;
2 weeks
relative weight;
6 weeks
relative weight;
14 weeks
mg/kg-day (F)
absolute weight;
2 weeks
absolute weight;
6 weeks
absolute weight;
14 weeks
relative weight;
2 weeks
relative weight;
6 weeks
relative weight;
14 weeks
0
0
0
0
0
0
0
0
0
0
0
0
0
0
151
0
1
-4
3
6*
4
171
5
3
4
2
0
4*
381
5
-4
-1
6
3
8*
422
5
7
3
5
5
5*
960
17
5
18*
25*
19*
28*
1,069
11
11*
15*
19*
17*
21*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1997b)
Rat (F344); 15 males/group
0, 300, 2,800, 25,000 ppm
0, 20, 200, 2,200 mg/kg-day
Diet
10 weeks
NTP (1997b)
Rat (F344); 15 males/group
0, 300, 900, 2,800, 8,300,
25,000 ppm
0, 30, 60, 180, 550, "high"
mg/kg-dayc
Diet
26 weeks
NTP (1997b)
Rat (F344); 60/sex/group;
assessed in 10 rats/sex/group at
15-month interim sacrifice
0, 3,000, 6,000, 12,000 ppm
(males); 0, 6,000, 12,000,
24,000 ppm (females)
0, 120, 240, 500 mg/kg-day
(males); 0 300, 600,
1,200 mg/kg-day (females)
Diet
2 years
Tvl et al. (2004)
Rat (CD); 30 FO and Fl parental
rats/sex/group
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-day
Diet
Multigenerational study
Exposure 10 weeks prior to
mating and through mating,
gestation, and lactation periods
(females) or 21 days after
mating (males)
Results3
Liver weight (percent change compared to control)
mg/kg-day
absolute weight
relative weight
0
0
0
20 200
1 0
2 2
2,200
-24*
6*
Liver weight (percent change compared to control)
mg/kg-day
absolute weight
relative weight
0
0
0
30 60 180 550
7 13 3 17*
2 3 3 14*
ND
-3
42*
Liver weight, 15 months (percent change compared to control)
mg/kg-day (Mal)e
absolute weight
relative weight
mg/kg-day (F)
absolute weight
relative weight
0
0
0
0
0
0
120 240
4 0
4 7
300 600
2 8
2 6
500
2
12*
1,200
-3
26*
Liver weight (percent change compared to control)
mg/kg-day
0
50 250
750
Absolute weight
FO males
Fl males
FO females
Fl females
0
0
0
0
-1 8
1 10*
4 9
1 8
13*
5
15*
6
Relative weight
FO males
Fl males
FOfemales
0
0
0
1 4
0 6
4 9
16*
16*
19*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Nagao et al. (2000)
Rat (Sprague-Dawley);
25 sex/generation/group
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
FO males and females: exposure
for 12 weeks prior to mating,
2 weeks cohabitation, and until
necropsy at 23 weeks of age
(males) or postpartum day 22
(females); Fl animals: from
weaning until necropsy at
PND22
Asoetal. (2005)
Rat(Crj:CD(SD)IGS);
24 sex/generation/group
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
FO and Fl exposed for 4 weeks
prior to mating, through mating
for 10 weeks, and until weaning
of offspring (females) or
necropsy (males)
Monsanto (1983)
Rat (Sprague-Dawley);
25/sex/group; interim sacrifice
of 10 rats/sex/group at 7 weeks
0, 51, 218, 789 mg/m3
Inhalation (whole-body)
13 weeks
Results3
Fl females
0
0 4
6
Liver weight (percent change compared to control)
mg/kg-day
0
20 100
500
Absolute weight
FO males
Fl males
FO females
Fl females
0
0
0
0
-3 1
0 -8
1 2
0 -2
11*
0
6
1
Relative weight
FO males
Fl males
FO females
Fl females
0
0
0
0
-1 1
4 -1
2 2
-1 -3
20*
15*
5
0
Liver weight (percent change compared to control)
mg/kg-day
0
100 200
400
Absolute weight
FO males
Fl males
FOfemales
Fl females
0
0
0
0
5 9
5 5
3 12*
3 7
14
12
11*
10
Relative weight
FO males
Fl males
FOfemales
Fl females
0
0
0
0
3 5
5 8*
3 6*
-1 1
14*
18*
10*
8*
Liver weight (percent change compared to control)
mg/kg-day
0
51 218
789
Absolute weight
male (7 weeks)
female (7 weeks)
male (13 weeks)
female (13 weeks)
0
0
0
0
0 7
1 4
9 9
4 5
18*
17*
24*
11*
Relative weight
male (7 weeks)
0
0 10*
18*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1990)
Mouse (Swiss albino CD-I);
28-30 females/group (except
n = 14 in 4,121 group)
0, 182, 910, 2,330,
4,121 mg/kg-day
Diet
CDs 6-15
NTP (1997a)
Rat (F344); 50-60/sex/group;
interim sacrifice of
10 rats/sex/group at 15 months
0, 12,000 ppm (males); 0,
24,000 ppm (females)
0, 500 mg/kg-day (males); 0,
1,200 mg/kg-day (females)
Diet
4 exposure protocols: ad libitum
feeding, weight-matched
controls, restricted feed
(2 years), and restricted feed
(lifetime)
2 years to lifetime
Piersma et al. (2000)
Rat (Harlan Cpb-WU);
10 females/group
0, 270, 350, 450, 580, 750, 970,
1,250, 1,600, 2,100 mg/kg-day
Gavage
Results3
female (7 weeks) 01 1
male (13 weeks) 07 6
female (13 weeks) 04 5
13*
21*
12*
Liver weight, GD 17 (percent change compared to control)
mg/kg-day 0 182 910 2,330
absolute weight 00-1 -15*
relative weight 002 26*
4,121
NE
NE
Liver weight, ad libitum and weight-matched (percent change compared to
control
mg/kg-day (M) 0 (ad 500 (ad 0 (weight-matched)
libitum) libitum)
absolute weight 02 0
relative weight 0 12* 0
mg/kg-day (F) 0 (ad 1,200 (ad 0 (weight-matched)
libitum) libitum)
absolute weight 0-3 0
relative weight 0 26* 0
500 (weight-
matched)
20*
22*
1,200
(weight-
matched)
64*
51*
Feed-restricted 2 years or lifetime (percent change compared to control)
mg/kg-day (M) 0 500
absolute weight 0 6
relative weight 0 11*
mg/kg-day (F) 0 1,200
absolute weight 0 -1
relative weight 0 20*
Liver weight (percent change compared to control)
mg/kg- Q 27Q 35Q 45Q 5go 75Q g70 Q Q
day
2,100
Relative liver weightd
0 8 6 6 6 11 11 19 13
22
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
GD 6-15 or 6-20
Ahmad et al. (2014)
Rat (Albino); PO, female
(6/group)
0, 4, 20, 100 mg/kg
Gavage
GD 14 to parturition
TNO (1998a)
Rat (Wistar); PO, female
(28/group)
0, 100, 1,000, 3,000 u.g/L
(equivalent to 0.016, 0.171,
0.489 mg/kg-day, average of
reported intake over premating,
gestation, and lactation)
Drinking water
FO females: 2 weeks prior to
mating, through mating,
gestation, and lactation; FO
males: during mating; Fl
animals were not treated after
weaning
Results3
short
exposure
(CDs
6-15)
long
exposure
(GDs
6-20)
Note: No data with respect to absolute liver weight were provided
authors.
30
by study
Liver weight (percent change compared to control)
mg/kg-day 04 20
Fl male absolute 0 -3 -3
liver weight
100
-6
Liver weight (percent change compared to control)
mg/kg-day 0 0.016 0.171
0.489
Absolute weight
Fl female 03 4
Fl male 00 4
2
0
Relative weight
Fl female 01 2
Flmale 01 3
1
1
Liver histopathology
NTP (1989)
Rat (Sprague-Dawley CD);
30 females/dose
0, 420, 1,100, 1,640 mg/kg-day
Diet
CDs 6-15
No histopathological effects were observed in the livers of control
dams (10/group); other groups were not examined.
or high-dose
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
BIBRA (1978)
Rat (Wistar); 27/sex/dose or
45/sex/group (control)
0, 2,000, 5,000, 12,000 ppm
0, 151, 381, 960 mg/kg-day
(males); 0,171,422,
1,069 mg/kg-day (females);
interim sacrifices of
9 controls/sex/group and
6 treated rats/sex/group at
2 and 6 weeks
Diet
14 weeks
Results3
Percent incidence
mg/kg-day (M) 0
151
281
960
Individual cell or focal necrosis
2 weeks 0
6 weeks 0
14 weeks 11
33
17
7
0
0
13
0
83*
50*
Inflammatory cells
2 weeks 11
6 weeks 78
14 weeks 78
17
17*
80
67*
67
80
0
67
71
Bile duct hyperplasia
6 weeks 0
14 weeks 4
17
7
33
27
33
0
Portal vacuolation/fatty change
2 weeks 0
14 weeks 11
0
0
0
7
0
0
Occasional foci of hemorrhage
14 weeks 0
0
7
0
Percent incidence
mg/kg-day (F) 0
171
422
1,069
Individual cell or focal necrosis
2 weeks 0
6 weeks 0
14 weeks 0
0
0
13
0
0
0
0
0
0
Inflammatory cells
2 weeks 67
6 weeks 89
14 weeks 56
50
50
67
33
33*
73
50
67
53
Bile duct hyperplasia
6 weeks 0
14 weeks 0
0
0
0
0
0
0
Portal vacuolation/fatty change
2 weeks 11
0
0
0
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1997b)
Rat (F344); 15 males/group
0, 300, 2,800, 25,000 ppm
0, 20, 200, 2,200 mg/kg-day
Diet
10 weeks
NTP (1997b)
Rat (F344); 15 males/dose
0, 300, 900, 2,800, 8,300,
25,000 ppm
0, 30, 60, 180, 550, "high"
mg/kg-dayc
Diet
26 weeks
NTP (1997b)
Rat (F344); 60/sex/group
0, 3,000, 6,000, 12,000 ppm
(males); 0, 6,000, 12,000,
24,000 ppm (females)
0, 120, 240, 500 mg/kg-day
(males); 0 300, 600,
1,200 mg/kg-day (females)
Diet
2 years
Tvl et al. (2004)
Rat (CD); 30 FO Fl parental
rats/sex/dose
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-day
Diet
Multigenerational study
Results3
14 weeks 11 7
0 0
Occasional foci of hemorrhage
14 weeks 0 0
0 0
No significant effects were reported by study authors in control or high-dose
animals (quantitative data not shown).
No significant effects were reported by study authors in control or high-dose
animals (quantitative data not shown).
Percent incidence at study termination
mg/kg-day (M) 0 120
granuloma 0 0
mg/kg-day (F) 0 300
cytoplasmic 14 12
vacuolization of
hepatocytes
Note: At 2 years, the incidences of granulomas
vacuolization of hepatocytes (in females) were
the study authors.
240 500
0 14
600 1,200
4 0
(in males) and cytoplasmic
not considered significant by
Percent incidence at study termination
mg/kg-day 0 50
250 750
Histopathological lesions3
FO males 0 0
Fl males 0 0
FOfemales 3 0
0 28
0 0
7 30
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Exposure 10 weeks prior to
mating and through mating,
gestation, and lactation periods
(females) or through 21 days
after end of mating (males)
Nagao et al. (2000)
Rat (Sprague-Dawley);
25 sex/generation/group;
assessed in 10 control and high-
dose rats/sex/group
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
FO males and females: exposure
for 12 weeks prior to mating,
2 weeks cohabitation, and until
necropsy at 23 weeks of age
(males) or postpartum day 22
(females); Fl animals: exposure
from weaning until necropsy
Asoetal. (2005)
Rat(Crj:CD(SD)IGS);
24 sex/generation/group
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
FO and Fl exposed for 4 weeks
prior to mating, through mating
for 10 weeks, and until weaning
of offspring (females) or
necropsy (males)
Monsanto (1983)
Rat (Sprague-Dawley);
25/sex/group; interim sacrifice
of 10 rats/sex/group at 7 weeks
0, 51, 218, 789 mg/m3
Inhalation (whole-body)
13 weeks
Results3
Fl females 00 0 17
Percent incidence
mg/kg-day 0 20 100 500
Fatty change; periportal
FO males 30 NE NE 0
Fl males 20 NE NE 0
Fibrosis; capsule/subcapsule; diaphragmatic nodule
FO females 10 NE NE 0
Granulation; subcapsule; focal
FO females 10 NE NE 0
Fl females 10 NE NE 0
No significant treatment-related effects were observed by the study authors in
FO or Fl parental males or females (quantitative data not reported).
Percent incidence
mg/kg-day (M) 0 51 218 789
tiny granulomas, 20 30 20 0
7 weeks
tiny granulomas, 70 7 7
13 weeks
focal necrosis, 00 0 0
7 weeks
lymphoid focus, 00 10 0
7 weeks
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1990)
Mouse (Swiss albino CD-I);
28-30 females/group (except
n = 14 in 4,121 group); assessed
in 10 dams/group (except the
high-dose)
0,182,910,2,330,4,121
mg/kg-day
Diet
GDs 6-15; necropsy at GD 17
NTP (1997a)
Rat (F344); 50-60/sex/group;
interim sacrifice of
10 rats/sex/group at 15 months
0, 12,000 ppm (males); 0,
24,000 ppm (females)
0, 500 mg/kg-day (males); 0,
1,200 mg/kg-day (females)
Diet
4 exposure protocols: ad libitum
feeding, weight-matched
controls, restricted feed
(2 years), and restricted feed
(lifetime)
2 years to lifetime
Results3
mg/kg-day (F)
tiny granulomas, 30 20 10
7 weeks
tiny granulomas, 07 0
13 weeks (focal)
focal necrosis, 0 10 0
7 weeks
lymphoid focus, 00 0
7 weeks
No significant treatment-related effects were observed by study
(quantitative data not reported).
Percent incidence (ad libitum and weight-matched protocols)
mg/kg-day (M) 0 0
(ad libitum) (weight-matched)
15 months
basophilic focus 60 20
granuloma 0 0
inflammation; 0 10
subacute
hepatocyte; 20 30
cytoplasmic
vacuolization
lobules, necrosis 0 0
2 years
basophilic focus 44 40
granuloma 0 0
inflammation; 0 0
subacute
hepatocyte; 12 18
cytoplasmic
vacuolization
0
7
0
0
authors
500
10
10
10
0
20
28
14
8
8
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results3
lobules, necrosis 4 4
2
Percent incidence (ad libitum and weight-matched protocols)
mg/kg-day (F) 0 (ad libitum) 0 (weight-matched)
1,200
15 months
basophilic focus 100 70
granuloma 10 20
90
0
2 years
centrilobular; 2 4
necrosis
hepatocyte; 14 2
vacuolization
cytoplasmic
lobules, necrosis 12 2
0
0
12
Percent incidence (Feed-restricted; 2 years or lifetime protocols)
mg/kg-day (M) 0 500 0
(2 years) (2 years) (lifetime)
500
(lifetime)
15 months
basophilic focus 10 0 NA
granuloma 0 10 NA
hepatocyte; 20 0 NA
vacuolization
cytoplasmic
NA
NA
NA
At study termination (2 years or lifetime)
basophilic focus 32 30 14
granuloma 20 2
inflammation; 44 0
subacute
hepatocyte; 80 4
vacuolization
cytoplasmic
lobules, necrosis 26 10
24
6
0
8
10
Percent incidence (Feed-restricted; 2 years or lifetime protocols)
mg/kg-day (F) 0 1,200 0
(2 years) (2 years) (lifetime)
1,200
(lifetime)
15 months
basophilic focus 100 30 NA
granuloma 10 0 NA
NA
NA
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1982)
F344 rats; 50 sex/group
0, 6,000, 12,000 ppm
0, 474, 947 mg/kg-day (males);
0, 550, 1,100 mg/kg-day
(females)
Diet
28 weeks (males) or 103 weeks
(females)
NTP (1982)
B6C3Fi mice; 50 sex/group
0, 6,000, 12,000 ppm
0, 474, 947 mg/kg-day (males);
0, 550, 1,100 mg/kg-day
(females)
Diet
103 weeks
Results3
inflammation; 10 0 NA
subacute
NA
At study termination (2 years or lifetime)
basophilic focus 80 82 64
granuloma 16 16 22
inflammation; 66 2
subacute
centhlobular; 00 2
necrosis
hepatocyte; 02 12
vacuolization
cytoplasmic
lobules, necrosis 12 4 8
76
10
0
0
0
8
Percent incidence
mg/kg-day (F) 0 550
necrosis; NOS 0 2
necrosis; focal 4 0
necrosis; diffuse 0 2
basophilic cyto 88 69
change
Note: Males were not examined histopathologically.
1,100
0
4
0
70
Percent incidence at study termination
mg/kg-day (M) 0 474
necrosis; focal 2 0
necrosis; 0 2
hemorrhagic
mg/kg-day (F) 0 550
necrosis; NOS 0 0
necrosis; focal 2 0
947
0
0
1,100
2
0
1
2
3
4
5
6
7
8
9
*Statistically significant (p <0.05) relative to controls based on statistics performed by the study authors.
aPercent change compared to control calculated as 100 x ([treated value - control value] 4- control value).
bAII studies reported relative weight in addition to absolute weight; where patterns were similar only relative
weight is included in this table.
°The high-dose group corresponds to 25,000 ppm BBP; a reliable estimate of dose could not be calculated. The
study authors estimated doses for all but the high-dose group based on measured body weights and food
consumption. Food consumption was not measured in the 25,000 ppm BBP group due to excessive scattering of
feed, and because the mean body weight of this group was 30% lower than controls.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 d Values reported by the study authors were estimated from published graphs using "Grab It!", a Microsoft Excel
2 based free software application used to digitizes data from image files. Publisher: www.datatrendsoftware.com.
3
4 eHistopathology reported by (Tyl et al. (2004)) includes: subtle to slight changes including diffuse cytomegaly,
5 variable karyomegaly, reduced cytoplasmic glycogen, increased cytoplasmic eosinophilia (increased numbers of
6 peroxisomes), increased cytoplasmic granularity.
7
8 F = female(s); M = male(s); ND = not determined; NE = not examined; NA = not applicable, NOS = not otherwise
9 specified
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
NTP, 1997b; lOwks; male F344rats
BIBRA, 1978; 14wks; male Wistarrats
BIBRA, 1978; 14wks; female Wistarrats
NTP, 1997h; 26wks; male F344rats
NTP 1989; CD 6-15; PO female S-D rats
TNO, 1998a; 2-gen; Fl male & female Wistarrats
Asocial., 2005; 2-gen; Fl males & FO female S-D rats
Asoetal., 2005; 2-gen; FO males & Fl female S-D rats
Nagao etal,, 2000; 2-gen; FO, Fl male S-D rats
Nagaoetal., 2000; 2-gen; FO, Fl female S-D rats
Tyl etal., 2004; 2-gen; Fl female CD rats
Tyl etal., 2004; 2-gen; FO, Fl male & FO female CD rats
NTP, 1997b; 2 yrs; male F344 rats
NTP, 1997h; 2 yrs; female F344 rats
NTP, 1997a; lifetime ad-lib feed; male F344 rats
NTP, 1997a; lifetime ad-lib feed; female F344 rats
NTP, 1990; CD 6-15; CD-I mice
NTP, 1997b; lOwks; male F344rats (decreased liver wt)
BIBRA, 1978; 14wks; male Wistarrats
BIBRA, 1978; 14wks; female Wistarrats
NTP, 1997b; 26wks; male F344rats
NTP 1989; CD 6-15; PO female S-D rats
Ahmad, 2014; CD 14-parturition; Fl male albino rats
TNO, 1998a; 2-gen; Fl male & female Wistar rats
Asoetal., 2005; 2-gen; FO.F1 male and Fl female S-D rats
Asoetal., 2005; 2-gen; FO female S-D rats
Nagao etal., 2000; 2-gen; FO male S-D rats
Nagao etal.,2000; 2-gen; Fl male &FO, Fl female S-D rats
Tyl etal., 2004; 2-gen; Fl female CD rats
Tyl etal., 2004; 2-gen; FO male & female CD rats
Tyl etal., 2004; 2-gen; Fl male CD rats
NTP, 1997b; 2 yrs; male F344 rats
NTP, 1997b; 2 yrs; female F344 rats
NTP, 1997a; ad lib wt-matched; male F344 rats
NTP, 1997a; ad lib feed restricted male F344 rats
NTP, 1997a; ad lib wt-matched female F344 rats
NTP, 1997a; ad lib feed restricted female F344 rats
NTP, 1990; CD 6-15; CD-I mice (decreased liver wt)
• = statistically significant
n - not statistically significant
0.01
Q
-B Q
0.1
10 100 1000
10000
Dose (rng/kg-day)
2
3
Figure 3-13. Exposure-response array of liver weight effects following oral
exposure to BBP.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
2
3
NTP, 1997b; lOwks; male F344rats
BIBRA, 1978; 14wks; female Wistarrats
BIBRA, 1978; 14wks; necrosis; male Wistar rats
NTP, 1997b; 26wks; male F344rats
NTP 1989; CD 6-15; PO female S-D rats
Aso etal., 2005; 2-gen; male & female S-D rats
Nagao etal., 2000; 2-gen; male & female S-D rats
<
n>
~i
X Tyl etal., 2004; 2-gen; CD rats
w
rt-
o
"O
3- NTP, 1997a; 15 mo.; ad lib wt matched male F344 rats
o_
o"
29
NTP, 1997a; 15 mo.; ad lib wt matched female F344 rats
NTP, 1997b; 2 yrs; male F344 rats
NTP, 1997b; 2 yrs; female F344 rats
NTP, 1982; 103wks; female F344 rats
NTP,1990;GD6-15;CD-1 mice
NTP, 1982; 103 wks; male B6C3F1 mice
NTP, 1982; 103wks; female B6C3F1 mice
- statistically significant
- not statistically significant
10
Q B B
B B El
Q B B
100
1000
Dose (mg/kg-day)
10000
Figure 3-14. Exposure-response array of liver histopathological effects
following oral exposure to BBP.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.3.5. Kidney Effects
2
3
Table 3-27. Evidence pertaining to kidney effects in animals following oral
and inhalation exposure to BBP
Reference and study design
Results
Kidney weight°'b
BIBRA (1978)
Rat (Wistar); 27/sex/dose or
45/sex/group (control); interim
sacrifices of 9 controls/sex/group
and 6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
0, 151, 381, 960 mg/kg-day
(males);
0, 171, 422, 1,069 mg/kg-day
(females)
Diet
14 weeks
NTP (1997b)
Rat (F344); 15 males/group
0, 300, 2,800, 25,000 ppm
0, 20, 200, 2,200 mg/kg-day
Diet
10 weeks
Kidney weight (percent change compared to control)
mg/kg-day (M)
absolute weight, 2
wee s
absolute weight, 6
weeks
absolute weight, 14
weeks
relative weight, 2
WGsks
relative weight, 6
weeks
relative weight, 14
weeks
mg/kg-day (F)
absolute weight, 2
weeks
absolute weight, 6
weeks
absolute weight, 14
weeks
relative weight, 2
weeks
relative weight, 6
weeks
relative weight, 14
weeks
Right kidney weight
mg/kg-day
absolute weight
relative weight
0
0
0
0
0
0
0
0
0
0
0
0
0
0
151
3
6
7*
6
171
7
3
3
381
6
7*
7*
8*
422
6
3
8*
960
1
6
8*
12*
1,069
5
1 ")*
J.Z
6
1 /Tf*
J.U
19*
(percent change compared to control)
0
0
0
20
-2
-1
200
3
5
2,200
-25*
6
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1997b)
Rat (F344); 15 males/group
0, 300, 900, 2,800, 8,300,
25,000 ppm
0, 30, 60, 180, 550, "high"
mg/kg-dayd
Diet
26 weeks
NTP (1997b)
Rat (F344); 60/sex/group; assessed
in 10 rats/sex/group at 15-month
interim sacrifice
0, 3,000, 6,000, 12,000 ppm
(males); 0, 6,000, 12,000,
24,000 ppm (females)
0, 120, 240, 500 mg/kg-day
(males); 0 300, 600,
1,200 mg/kg-day (females)
Diet
2 years
Tvl et al. (2004)
Rat (CD); 30 FO and Fl parental
rats/sex/group
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-day
Diet
Multigenerational study
Exposure 10 weeks prior to mating
and through mating, gestation,
and lactation (females) or through
21 days after end of mating
(males)
Results
Right kidney weight
mg/kg-day
absolute weight
relative weight
Right kidney weight,
mg/kg-day (M)
absolute weight
relative weight
mg/kg-day (F)
absolute weight
relative weight
(percent change compared to control)
0 30 60 180 550
007-2 11
0 -5 -2 -3 8
ND
-20*
18*
15 months (percent change compared to control)
0 120 240
0 10 4
0 9* 10*
0 300 600
08 9*
0
8 7
500
6
16*
1,200
-7
21*
Kidney weight (percent change compared to control)
mg/kg-day
0 50 250
750
Absolute weight
FO males
Fl males
FOfemales
Fl females
0 -3 7*
0 3 12*
02 6*
02 8*
8*
-4
6
6
Relative weight
FO males
Fl males
FOfemales
Fl females
0-1 3
02 7*
02 6*
01 5
10*
5
9*
4
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Nagao et al. (2000)
Rat (Sprague-Dawley);
25 sex/generation/group
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
FO males and females: exposure
for 12 weeks prior to mating,
2 weeks cohabitation, and until
necropsy at 23 weeks of age
(males) or postpartum day 22
(females); Fl animals: exposure
from weaning until necropsy
Asoetal. (2005)
Rat(Crj:CD(SD)IGS);
24 sex/generation/group
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
FO and Fl exposed for 4 weeks
prior to mating, through mating
for 10 weeks, and until weaning of
offspring (females) or necropsy
(males)
Results
Kidney weight (percent change compared to control)
mg/kg-day
0
20
100
500
Absolute weight
FO males
Fl males
FOfemales
Fl females
0
0
0
0
-3
0
3
1
2
1
7*
6
7
4
7*
4
Relative weight
FO males
Fl males
FOfemales
Fl females
0
0
0
0
-2
4
5
0
2
9*
8*
5
14
18*
6*
5
Kidney weight (percent change compared to control)
mg/kg-day
0
100
200
400
Absolute weight
FO males left kidney
Fl males left kidney
FOfemales left
kidney
Fl females left
kidney
FO males right
kidney
Fl males right
kidney
FOfemales right
kidney
Fl females right
kidney
0
0
0
0
0
0
0
0
6
-1
17
6
4
0
3
9
8*
0
12*
7
7
0
11*
12
9*
0
12*
6
8
0
8*
5
Relative weight
FO males left kidney
Fl males left kidney
FOfemales left
kidney
0
0
0
3
0
6
7
0
6
10*
3
12*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Monsanto (1983)
Rat (Sprague-Dawley);
25/sex/group; interim sacrifice of
10 rats/sex/group at 7 weeks
0, 51, 218, 789 mg/m3
Inhalation (whole-body)
13 weeks
Results
Fl females left 0
kidney
FO males right 0
kidney
Fl males right 0
kidney
FO females right 0
kidney
Fl females right 0
kidney
3
3
0
3
3
3
3
0
6
6
6
10
3
6*
3
Kidney weight (percent change compared to control)
mg/kg-day 0
51
218
789
Absolute weight, left kidney
males, 7 weeks 0
females, 7 weeks 0
males, 13 weeks 0
females, 13 weeks 0
2
-2
5
3
8
8
8
6
22*
13
18*
14*
Absolute weight, right kidney
males, 7 weeks 0
females, 7 weeks 0
males, 13 weeks 0
females, 13 weeks 0
1
0
1
4
11*
10
7
7
21*
12*
17*
12*
Relative weight, left Kidney
males, 7 weeks 0
females, 7 weeks 0
males, 13 weeks 0
females, 13 weeks 0
3
-2
3
3
11*
4
5
7
22*
10
15*
15*
Relative weight, right kidney
males, 7 weeks 0
females, 7 weeks 0
males, 13 weeks 0
females, 13 weeks 0
1
0
0
4
13*
6
4
8
20*
9
14*
13*
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1989)
Rat (Sprague-Dawley CD);
30 females/group
0, 420, 1,100, 1,640 mg/kg-day
Diet
CDs 6-15
NTP (1990)
Mouse (Swiss albino CD-I); 28-30
females/group (except n = 14 in
4,121 mg/kg-day group)
0, 182, 910, 2,330, 4,121 mg/kg-
day
Diet
CDs 6-15
NTP (1997a)
Rat (F344); 50-60/sex/group;
interim sacrifice of
10 rats/sex/group at 15 months
0, 12,000 ppm (males); 0, 24,000
ppm (females)
0, 500 mg/kg-day (males); 0,
1,200 mg/kg-day (females)
Diet
4 exposure protocols: ad libitum
feeding, weight-matched controls,
restricted feed (2 years), and
restricted feed (lifetime)
Diet
2 years
Results
Kidney weight (percent change compared to control)
mg/kg-day
(Maternal)
0 420
1,100
16,40
Absolute weight
left kidney
right kidney
0 1
0 2
2
2
2
2
Relative weight
left kidney
right kidney
Right kidney weight,
mg/kg-day
absolute weight
relative weight
0 0
0 0
7
3
20*
16*
GD 17 (percent change compared to control)
0 182 910
0 0 5
008
2,330
10
62*
4,121
NE
NE
Kidney weight, ad libitum and weight-matched (percent change compared to
control)
mg/kg-day (M)
absolute weight
relative weight
mg/kg-day (F)
absolute weight
relative weight
500
0 (ad
(ad libitum) libitum)
0 6
0 16*
1,200
0 (ad
(ad libitum) libitum)
0 -6
0 22*
0
(weight
matched)
0
0
0
(weight
matched)
0
0
Kidney weight, Feed-restricted; 2 years or lifetime (percent change
to control)
mg/kg-day (M)
absolute weight
relative weight
0
0
0
500
12*
17*
500
(weight
matched)
20*
15*
1,200
(weight
matched)
30*
20*
compared
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Piersma et al. (2000)
Rat (Harlan Cpb-WU);
10 females/group
1, 270, 350, 450, 580, 750,
970,1,250, 1,600, 2,100 mg/kg-day
Gavage
GDs 6-15 or 6-20; dams sacrificed
onGD21
Ahmad et al. (2014)
Rat (Albino);
PO, female (6/group)
0, 4, 20, 100 mg/kg
Gavage
GD 14 to parturition
TNO (1998a)
Rat (Wistar); PO, female (28/group)
0, 100, 1,000, 3,000 u.g/L
(equivalent to 0.016, 0.171, 0.489
mg/kg-day, average of reported
intake over premating, gestation,
and lactation)
Drinking water
FO females: 2 weeks prior to
mating, through mating, gestation,
and lactation; FO males: during
mating; Fl animals were not
treated after weaning
Results
mg/kg-day (F) 0 1200
absolute weight 0 -4
relative weight 0 16*
Percent change compared to control
mg/kg-day 0 270 350 450 580 750 970 1,250 1,600 2,100
Relative kidney weight0
short 0214 10 10 16 20 19 33
exposure
(GDs 6-15)
long 0-1-32 6 11 14 20 36 24
exposure
(GDs 6-20)
Note: No data with respect to absolute kidney weight were provided by
study authors.
Kidney weight (percent change compared to control)
mg/kg-day 04 20 100
Fl male absolute 0 -1 -1 -11*
weight
Kidney weight (percent change compared to control)
mg/kg-day 0 0.016 0.160 0.481
Absolute weight
Fl female 0-311
Flmale 0-241
Relative weight
Fl female 0 -4* -1 0
Flmale 0-132
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Kidney histopathology
NTP (1997b)
Rat (F344); 60/sex/dose; assessed
in 10 rats/sex/group at 15-month
interim sacrifice and 50
rats/sex/group at study
termination
0, 3,000, 6,000, 12,000 ppm
(males); 0, 6,000, 12,000, 24,000
ppm (females)
0, 120, 240, 500 mg/kg-day
(males); 0 300, 600, 1,200 mg/kg-
day (females)
Diet
2 years
Nagao et al. (2000)
Rat (Sprague-Dawley);
25 sex/generation/group; assessed
in 10 control and high-dose
rats/sex
Results
Percent incidence
mg/kg-day (M) 0 120
nephropathy, 15
months 100 100
renal tubule; 100 100
pigmentation, 15
months
nephropathy, 2 96 94
years
renal tubule; 98 96
pigmentation, 2
years
mineralization, 20 2
years
transitional 12 20
epithelium
hyperplasia, 2 years
mg/kg-day (F) 0 300
nephropathy, 15 70 100
months
renal tubule; 100 100
pigmentation, 15
months
mineralization, 15 100 90
months
nephropathy, 2 68 94*
years
renal tubule; 98 98
pigmentation, 2
years
mineralization, 2 86 68*
years
transitional 0 6
epithelium
hyperplasia, 2 years
Percent incidence
mg/kg-day (M) 0 20
Basophilic tubule in cortex
240 500
100 90
100 100
100 96
100 100
4 0
12 2
600 1,200
100 100
100 100
90 80
86* 90*
98 94
74 70*
14* 8
100 500
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
FO males and females: exposure
for 12 weeks prior to mating, 2
weeks cohabitation, and until
necropsy at 23 weeks of age
(males) or postpartum day 22
(females); Fl animals: exposure
from weaning until necropsy
Results
FO
Fl
NA
100
NE
NE
NE
NE
NA
100*
Cast, cortex/medulla
FO
Fl
20
20
NE
NE
NE
NE
40
40
Eosinophilic bodies
FO
Fl
70
30
NE
NE
NE
NE
50
10
Mineralization
FO
Fl
30
40
NE
NE
NE
NE
0
30
Cyst; medulla
FO
10
NE
NE
0
Degeneration; vacuolar, with hyaline droplet; proximal tubular epithelium
FO
10
NE
NE
0
Cellular infiltration, lymphocyte, interstitium
Fl
0
NE
NE
10
Dilatation, renal pelvis
Fl
10
NE
NE
10
Fibrosis; focal, subcapsule
Fl
mg/kg-day (F)
10
0
NE
20
NE
100
0
500
Basophilic tubule in cortex
FO
Fl
20
40
NE
NE
NE
NE
50
50
Fibrosis; focal, subcapsule
FO
10
NE
NE
0
Mineralization; papilla
Fl
20
NE
NE
0
Dilatation; renal pelvis; right side
Fl
10
NE
NE
0
Dilatation, collecting tubule, medulla, and papilla
Fl
0
NE
NE
10
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Hotchkissetal. (2004)
Rat (Sprague-Dawley);
6 litters/group
0, 500 mg/kg-day
Gavage
CDs 14-18
BIBRA (1978)
Rat (Wistar); 27/sex/dose or
45/sex/group (control); interim
sacrifices of 9 controls/sex/group
and 6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
0, 151, 381, 960 mg/kg-day
(males)0
0, 171, 422, 1,069 mg/kg-day
(females)
Diet
14 weeks
Results
Percent incidence
mg/kg-day
in male offspring
0
at 3 months of age
hydronephrosis 3
Note: Individual percent incidence was calculated on a
included both left and right tissues.
500
30*
per animal
basis and
Percent incidence
mg/kg-day (M)
0
151
381
960
Early nephrosis
2 weeks
6 weeks
14 weeks
33
67
33
33
17
67
17
17
33
0
17
14
Basophilia and tubular hyperplasia
2 weeks
6 weeks
14 weeks
44
0
26
50
33
0
33
83*
0
17
33
43
Foci of inflammatory cells
2 weeks
6 weeks
14 weeks
Foci of calcium at
14 weeks
0
0
0
17
17
7
0
0
0
0
0
7
co rticomedu Mary junction
0
0
0
0
Transitional cell hyperplasia
14 weeks
mg/kg-day (F)
0
0
7
171
0
422
0
1,069
Early nephrosis
2 weeks
6 weeks
14 weeks
0
0
7
0
0
0
17
0
0
17
0
0
Basophilia and tubular hyperplasia
2 weeks
6 weeks
33
11
50
17
33
0
33
0
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1997a)
Rat (F344); 50-60/sex/group;
interim sacrifice of 10
rats/sex/group at 15 months
0, 12,000 ppm (males); 0,
24,000 ppm (females)
0, 500 mg/kg-day (males); 0,
1,200 mg/kg-day (females)
Diet
4 exposure protocols: ad libitum
feeding, weight-matched controls,
restricted feed (2 years), and
restricted feed (lifetime)
2 years to lifetime
Results
14 weeks 0 7 13
0
Foci of inflammatory cells
2 weeks 000
6 weeks 000
14 weeks 000
0
17
0
Foci of calcium at corticomedu Mary junction
14 weeks 000
7
Transitional cell hyperplasia
14 weeks 000
0
Percent incidence
Ad libitum and weight-matched
mg/kg-day (M) 0 (ad libitum) 0 (weight
matched)
nephropathy, 15 100 90
months
inflammation; 0 2
suppurative, 2
years
mineralization, 20 2
years
nephropathy, 2 96 96
years
transitional 12 0
epithelium;
hyperplasia, 2 years
mg/kg-day (F) 0 (ad libitum) 0 (weight
matched)
mineralization, 15 100 90
months
nephropathy, 15 70 20
months
hydronephrosis, 20 4
years
mineralization, 2 86 98
years
nephropathy, 2 68 64
years
500
90
8
0
96
2
1,200
80
100
2
70
90
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Monsanto (1983)
Rat (Sprague-Dawley);
25/sex/group; interim sacrifice of
10 rats/sex/group at 7 weeks
0, 51, 218, 789 mg/m3
Results
transitional 0 8
epithelium;
hyperplasia, 2 years
8
Feed-restricted; 2 years or lifetime
mg/kg-day (M) 0 500 0
(2 years) (2 years) (lifetime)
mineralization, 15 0 10 NA
months
nephropathy, 15 80 100 NA
months
hydronephrosis, 20 00
years
mineralization, 2 10 8 12
years
nephropathy, 2 86 92 98
years
transitional 2 44
epithelium;
hyperplasia, 2 years
500
(lifetime)
NA
NA
2
10
98
2
Feed-restricted; 2 years or lifetime
mg/kg-day (F) 0 1,200 0
(2 years) (2 years) (lifetime)
mineralization, 15 100 80 NA
months
nephropathy, 15 100 90 NA
months
hydronephrosis, 20 20
years
inflammation; 0 00
suppurative
mineralization, 2 92 68 90
years
transitional 2 40 4
epithelium;
hyperplasia, 2 years
1,200
(lifetime)
NA
NA
0
4
62
58
Percent incidence
mg/kg-day (M) 0 51 218
789
Focal scar
6 weeks 10 0 10
0
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Inhalation (whole-body)
13 weeks
Results
Lymphoid foci
6 weeks
13 weeks
10
0
0
7
0
0
10
7
Tubular basophilia
6 weeks
13 weeks
20
7
0
0
0
0
0
0
Small cysts
6 weeks
13 weeks
0
0
0
7
0
0
0
0
Pelvic dilation
6 weeks
13 weeks
10
0
0
0
0
7
10
7
Tinygranuloma
13 weeks
0
0
7
0
Focal interstitial nephritis
13 weeks
0
0
0
7
Percent incidence
mg/kg-day (F)
0
51
218
789
Focal Scar
6 weeks
0
0
0
0
Lymphoid foci
6 weeks
13 weeks
0
0
0
7
10
0
0
0
Tubular basophilia
6 weeks
13 weeks
10
0
0
0
0
0
0
0
Small cysts
6 weeks
13 weeks
20
0
0
7
0
0
10
0
Pelvic dilation
6 weeks
13 weeks
10
7
0
0
0
0
10
0
Tinygranuloma
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1982)
B6C3Fi mice, 50/sex/group
0, 6,000, 12,000 ppm
0, 474, 947 mg/kg-day (males); 0,
550, 1,100 mg/kg-day (females)
Diet
103 weeks
NTP (1982)
F344 rats; 50 sex/group
0, 6,000, 12,000 ppm
0, 474, 947 mg/kg-day (males); 0,
550, 1,100 mg/kg-day (females)
Diet
28 weeks (males) or 103 weeks
(females)
Tvl et al. (2004)
Rat (CD); 30 FO and Fl parental
rats/sex/group
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-day
Diet
Multigenerational study
Results
13 weeks 0
0
0 0
Focal interstitial nephritis
13 weeks 0
0
0 0
Percent incidence
mg/kg-day (M) 0
mineralization 2
inflammation, 2
interstitial
nephropathy 0
mg/kg-day (F) 0
mineralization 0
inflammation, 2
interstitial;
nephropathy 2
tubule; 0
regeneration; NOS
474
0
2
2
550
0
2
0
2
947
0
21
0
1,100
2
6
0
0
Percent incidence
mg/kg-day (F) 0
mineralization 2
hydronephrosis 0
inflammation, 0
interstitial;
nephropathy 68
tubule; 2
regeneration; NOS
Note: Males were not examined
550
0
2
0
64
2
histopathologically.
1,100
2
0
2
40
0
No significant treatment-related effects were observed by study authors in
FO or Fl parental males or females (quantitative data not reported).
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Exposure 10 weeks prior to mating
and through mating, gestation,
and lactation (females) or through
21 days after end of mating
(males)
Asoetal. (2005)
Rat(Crj:CD(SD)IGS);
24 sex/generation/group
0,100, 200, 400 mg/kg-day
Gavage
Multigenerational study
FO and Fl exposed for 4 weeks
prior to mating, through mating
for 10 weeks, and until weaning of
offspring (females) or necropsy
(males)
No significant treatment-related effects were observed by study authors in
FO or Fl parental males or females (quantitative data not reported).
NTP (1990)
Mouse (Swiss albino CD-I); 28-30
females/group (except n = 14 in
4,121 mg/kg-day group); assessed
in 10 dams/group (except the high-
dose)
0,182, 910, 2,330, 4,121 mg/kg-
day
Diet
CDs 6-15
Histopathology at GD 17 was evaluated, but no significant treatment-related
effects were observed by study authors (quantitative data not reported).
NTP (1989)
Rat (Sprague-Dawley CD);
30 females/group
0, 420,1,100,1,640 mg/kg-day
Diet
CDs 6-15
Histopathology at GD 20 was evaluated, but no histopathological effects
were observed in the kidneys of control or high-dose dams (10/group); other
groups were not examined.
NTP U997b)
Rat (F344); 15 males/group
0, 300, 2,800, 25,000 ppm
0, 20, 200, 2,200 mg/kg-day
Diet
10 weeks
No significant effects reported by study authors in control or high-dose
animals (quantitative data not reported).
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
NTP U997b)
Rat (F344); 15 males/group
0, 300, 900, 2,800, 8,300, 25,000
ppm
0, 30, 60,180, 550, "high" mg/kg-
dayd
Diet
26 weeks
No significant effects reported by study authors in control or high-dose
animals (quantitative data not reported).
1
2
3
4
5
6
7
8
9
10
11
12
13
*Statistically significant (p <0.05) relative to controls based on statistics performed by the study authors.
aPercent change compared to control calculated as 100 x ((treated value - control value) -f control value).
bAII studies reported relative weight in addition to absolute weight; patterns were similar, and only relative weight
is included in this table.
c Values reported by the study authors were estimated from published graphs using "Grab It!", a Microsoft Excel
based free software application used to digitizes data from image files. Publisher: www.datatrendsoftware.com.
dThe high-dose group corresponds to 25,000 ppm BBP; a reliable estimate of dose could not be calculated. The
study authors estimated doses for all but the high-dose group based on measured body weights and food
consumption. Food consumption was not measured in the 25,000 ppm BBP group due to excessive scattering of
feed, and because the mean body weight of this group was 30% lower than controls.
NE = not examined; NOS = not otherwise specified
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
NTP, 1997b; lOwks; male F344 rats
BIBRA, 1978; 14wks; male Wistar rats
BIBRA, 1978; 14wks; female Wistar rats
NTP, 1997b; 26wks; male F344 rats
NTP 1989; GD 6-15; S-D rats
NTP, 1997b; 15 mo.; male F344 rats
TNO, 1998a male Wistar rats
TNO, 1998a; female Wistar rats
Aso etal., 2005; 2-gen; FO male & female S-D rats
Asocial., 2005; 2-gen; Fl male & female S-D rats
Nagao et al., 2000; 2-gen; FO male & Fl female S-D rats
Nagao etal., 2000; 2-gen; Fl males & FO female S-D rats
Tyl etal., 2004; 2-gen; FO male CD rats
Tyl etal., 2004; 2-gen; Fl male CD rats
Tyl etal., 2004; 2-gen; FO female CD rats
Tyl et al., 2004; 2-gen; Fl females CD rats
NTP, 1997b; 15 mo.; female F344 rats
NTP, 1997a; Males ad lib, wt matched, feed restricted F344 rats
NTP, 1997a; Female ad lib, wt matched, feed restricted F344 rats
NTP, 1990; CD 6-15; CD-I mice
NTP, 1997b; lOwks; male F344 rats
BIBRA, 1978; 14wks; female Wistar rats
BIBRA, 1978; 14wks; male Wistar rats
NTP, 1997b; 26wks; male F344 rats
NTP 1989; GD 6-15; S-D rats
Ahmad, 2014;GD 14-parturition;Fl male albino rats
Aso etal., 2005; 2-gen; FO male & female S-D rats
Aso etal., 2005; 2-gen; Fl male & female S-D rats
Nagao eta I., 2000; 2-gen; FO, Fl male&Fl female
Nagao etal,, 2000; 2-gen; FO female
Tyl etal., 2004; 2-gen; FO male & FO,F1 female CD rats
Tyl etal., 2004; 2-gen; Fl male CD rats
NTP, 1997b; 15 mo.; male F344 rats
NTP, 1997b; 15 mo.; female F344 rats
NTP, 1997a; males ad lib F344 rats
NTP, 1997a; males wt matched, feed restricted F344 rats
NTP, 1997a; females ad lib, feed restricted F344 rats
NTP, 1997a; females wt matched F344 rats
NTP, 1990; GD 6-15; female PO CD-I mice
D—B B D
B B B
B B-
B—B B B
B B El
• = statistically significant
3 = not statistically significant
0.01 0.1 1 10 100 1000 10000
Dose (mg/kg-day)
2
3
Figure 3-15. Exposure-response array of kidney weight effects following oral
exposure to BBP.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
NTP, 1997b; lOwks; male F344 rats
BIBRA, 1978; 14wks; renal performance; male Wistarrats
B1BRA, 1978; 14wks; male Wistarrats
BIBRA, 1978; 14wks; female Wistarrats
NTP, 1997b; 26wks; male F344 rats
NTP 1989; CD 6-15; S-D rats
Hotchkissetal., 2004; CD 14-18; hydronephrosismale S-D rats
Asoetal., 2005; 2-gen; FO or Fl parental S-D rats
Nagao etal., 2000; 2-gen; FO or Fl parental S-D rats
Tyl et al., 2004; 2-gen; FO or Fl parental CD rats
NTP, 1997a; 15 mo.; male F344 rats
NTP, 1997a; 15 mo.; female F344 rats
NTP, 1997b; 15 mo.; female F344 rats
NTP 1982; 103wks; nephropathy, inflammation, mineralization,
hydronephrosis female F344 rats
NTP, 1997b; 2 yr; transitional epithelial hyperplasia female F344 rats
NTP, 1997b; 2 yr; nephropathy, mineralization; female F344 rats
NTP, 1990; CD 6-15; female PO CD-I mice
NTP 1982; 103wks; male B6C3F1 mice
NTP 1982; 103wks; female B6C3F1 mice
2
3
= statistically significant
= not statistically significant
0.01 0.1 1 10
Dose (mg/kg-day)
100
1000
10000
Figure 3-16. Exposure-response array of kidney histopathological effects
following oral exposure to BBP.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.3,6. Pancreatic Effects
2 Table 3-28. Evidence pertaining to pancreatic effects in animals following oral
3 and inhalation exposure to BBP
Reference and study design
Results
Pancreas weight"
Tvl et al. (2004)
Rat (CD); 30 FO and Fl parental
rats/sex/group
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
Exposure 10 weeks prior to mating
and through mating, gestation, and
lactation (females) or through
21 days after end of mating (males)
Pancreas weight (percent change compared to control)
mg/kg-day
0 50
250
750
Absolute weights
FO males
Fl males
FOfemales
Fl females
0
-5
0 9
NR NR
NR NR
-7
14*
NR
NR
-2
14*
NR
NR
Relative weights
FO males
Fl males
FOfemales
Fl females
Note: No effect reported
0 -3
0 9
NR NR
NR NR
by study authors on
-10
9
NR
NR
1
25*
NR
NR
female pancreas weights
(quantitative data not reported).
Pancreas histopathology
BIBRA (1978)
Rat (Wistar); 27/sex/treatment group
or 45/sex/group (control)
0, 2,000, 5,000, 12,000 ppm
0, 151, 381, 960 mg/kg-day (males)";
0, 171, 422, 1,069 mg/kg-day
(females)b; interim sacrifices of
9 controls/sex/group and 6 treated
rats/sex/group at 2 and 6 weeks
Diet
14 weeks
Percent incidence
mg/kg-day (M)
focus ofexocrine
hyperplasia; 6 weeks
incidental pancreatic
lesion; 14 weeks
marginal islet
enlargement;
14 weeks
pancreatic lesions
mg/kg-day (F)
focus ofexocrine
hyperplasia; 6 weeks
0 151
0 NE
7 0
0 57*
0 0
0 171
11 NE
381
NE
7
20*
53*
422
NE
960
0
0
0
93*
1,069
0
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
incidental pancreatic
lesion; 14 weeks
marginal islet
enlargement; 14
weeks
pancreatic lesions, 14
weeks
0
NE
NE
NE
NE
NE
NE
0
Lesions include islet enlargement with cell vacuolation, peri-islet
congestion, peri-islet inflammatory cell infiltration, and slight fibrosis in
the endocrine pancreas; occasional pyknotic nuclei, acinar atrophy,
periacinar inflammatory cell infiltrate, and fibrosis observed less
frequently in the exocrine pancreas. The severity of these lesions
increased in a dose-related manner.
NTP U997b)
Rat (F344); 15 males/group
0, 300, 2,800, 25,000 ppm
0, 20, 200, 2,200 mg/kg-dayb
Diet
10 weeks
No significant effects reported by study authors in control or high-dose
animals (quantitative data not reported).
NTP U997b)
Rat (F344); 15 males/group
0, 300, 900, 2,800, 8,300,
25,000 ppm
0, 30, 60, 180, 550, "high"
mg/kg-dayb
Diet
26 weeks
No significant effects reported by study authors in control or high-dose
animals (quantitative data not reported).
NTP U997b)
Percent incidence
Rat (F344); 60/sex/group; assessed in
10 rats/sex/group at 15-month
interim sacrifice and
50 rats/sex/group at study
termination
0, 3,000, 6,000,12,000 ppm (males);
0, 6,000, 12,000, 24,000 ppm
(females)
0,120, 240, 500 mg/kg-day (males)";
0 300, 600,1,200 mg/kg-day
(females)"
mg/kg-day (M)
0
120
240
500
acinus hyperplasia 8
(severity") (2.5)
acinus adenoma 6
acinus carcinoma 0
acinus carcinoma or 6
adenoma
14(2.1) 18(2.3)
4
0
4
6
0
6
24 (2.3)
20
2
22
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Diet
2 years
NTP (1982)
B6C3Fi mice; 50/sex/group
0, 6,000, 12,000 ppm
0, 474, 947 mg/kg-day (males)"; 0,
550, 1,100 mg/kg-day (females)"
Diet
103 weeks
NTP (1982)
F344 rats; 50 sex/group
0, 6,000, 12,000 ppm
0, 474, 947 mg/kg-day (males)"; 0,
550, 1,100 mg/kg-day (females)"
Diet
28 weeks (males) or 103 weeks
(females)
Results
Females
acinus hyperplasia
(severity0)
acinus adenoma
0
2
(3.0)
0
300 600
8 4
(2.5) (2.5)
0 0
1,200
0
(-)
4
Percent incidence
mg/kg-day (M)
inflammation; NOS
inflammation; focal
atrophy; NOS
acinus; atrophy, focal
mg/kg-day (F)
dilatation/ducts
cystic ducts
inflammation; NOS
abscess; NOS
atrophy; NOS
acinus; atrophy, NOS
acinus; atrophy, focal
acinar-cell adenoma
leiomyosarcoma;
metastatic
0
0
0
0
0
0
2
0
0
0
0
5
0
0
0
474
0
2
2
2
550
0
2
2
2
2
0
0
0
2
974
2
0
2
0
1,100
0
2
0
0
0
0
2
2
0
Percent incidence
mg/kg-day (F)
inflammation; NOS
inflammation; focal
atrophy; NOS
acinus; atrophy, NOS
acinus; atrophy, focal
pancreatic islets;
islet-cell adenoma
Note: Males were not
0
0
0
0
2
6
0
examined
550
2
0
2
0
2
7
histopathologically.
1,100
0
2
2
0
2
0
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Monsanto (1983)
Rat (Sprague-Dawley); 25/sex/group;
interim sacrifice of 10 rats/sex/group
at 7 weeks
0, 51, 218, 789 mg/m3
Inhalation (whole-body)
13 weeks
NTP (1997a)
Rat (F344); 60/sex/group; interim
sacrifice of 10 rats/sex/group at
15 months
0, 12,000 ppm (males); 0,
24,000 ppm (females)
0, 500 mg/kg-day (males)b; 0,
1,200 mg/kg-day (females)b
Diet
Three studies: (1) ad libitum feeding
and weight-matched controls,
(2) restricted feed (2 years), and
(3) restricted feed (lifetime)
Diet
2 years to lifetime
Results
Percent incidence
mg/kg-day (M) 0 51 218
yellow pigment; peri- 700
islet; 13 weeks
periacinar round 000
cells; 13 weeks
mg/kg-day (f) 0 51 218
789
0
7
789
yellow pigment; peri- 0000
islet; 13 weeks
periacinar round 0000
cells; 13 weeks
Note: No significant histopathological effects were observed by study
authors in rats sacrificed at interim.
Percent incidence
Ad libitum and weight-matched
mg/kg-day (M) 0 (weight-
0 (ad libitum matched
control) control)
acinus; hyperplasia 8 4
acinus; adenoma 6 0
acinus; carcinoma 0 2
acinus; carcinoma or 6 2
adenoma
mg/kg-day (F) 0 (weight-
0 (ad libitum matched
control) control)
acinar hyperplasia 2 0
acinus, adenoma 0 0
500
24
20*
2
22
1,200
0
4
Feed restricted; 2 years
mg/kg-day (M) 0
acinus, focal
hyperplasia
acinus, adenoma 0
mg/kg-day (F) 0
500
6
0
1,200
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
acinar cell,
hyperplasia
acinus, adenoma 0
0
0
Feed- restricted, lifetime
mg/kg-day (M) 0
acinus hyperplasia 0
acinus, adenoma 0
mg/kg-day (F) 0
500
4
2
1,200
acinar cell,
hyperplasia
acinus, adenoma 0 2
There were no significant-treatment-related effects in females relative to
ad libitum or weight-matched controls or in males and females in the
restricted feed studies compared to their respective control groups.
1
2
3
4
5
6
7
8
*Statistically significant (p <0.05) relative to controls based on statistics performed by the study authors.
aPercent change compared to control calculated as 100 x ((treated value - control value) -f control value).
Calculated as follows: [% in diet x intake food/water (mg)] 4- body weight (kg) = mg/kg-day.
°Average severity in affected animals where 1 = minimal; 2 = mild; 3 = moderate; 4 = marked.
NE = not examined; NOS = not otherwise specified
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Tyl et a]., 2004; 2-gen
FO male abs & re! wt
Tyl et al., 2004; 2-gen
Fi male abs wt
Tyl et al., 2004; 2-gen
Fl male rel wt
NTP, 1997B; lOwks
male F344 rats
BIBRA,1978;14wks
female Wistar rats
B1BRA, 1978; 14wks; total pancreatic lesions
male Wistar rats
NTP, 1997b; 26wks
male F344 rats
n NTP1997B; 2yr; acinus hyperplasia, adenoma, carcinoma
w male F344 rats
£r NTP 1997b; 2yr; acinus hyperplasia, adenoma; female F344 rats
NTP 1982; 103wks
male B6C3F1 mice
NTP 1982; 103wks
female B6C3F1 mice
NTP 1997a; 2 yr ad-lib wt-matched control; acinus adenoma
male F344 rats
NTP 1997a; 2 yr ad-lib wt-matched control; female F344 rats
Q B O
Q B Q
Q B El
- statistically significant
= not statistically significant
10
100 1000
Dose (mg/kg-day)
10000
2
3
Figure 3-17. Exposure-response array of pancreatic effects following oral
exposure to BBP.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.3,7. Hematopoietic Effects
2
3
Table 3-29. Evidence pertaining to hematopoietic effects in animals following
oral and inhalation exposure to BBP
Reference and study design
Spleen weight"
Tvl et al. (2004)
Rat (CD); 30 FO and Fl parental
rats/sex/group; assessed in
54-86 male offspring/group and
43-87 female offspring/group
(>3 sex/litter/group if possible)
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
Exposure 10 weeks prior to mating
and through mating, gestation, and
lactation (females) or through
21 days after end of mating (males)
BIBRA (1978)
Rat (Wistar); 27/sex/group or
45/sex/group (control); interim
sacrifices of 9 controls/sex/group
and 6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
Diet: 0, 151, 381, 960 mg/kg-day
(males)"; 0, 171, 422,
1,069 mg/kg-day (females)b
3 months
Spleen weight, PND 21
mg/kg-day
Absolute weight
Fl males
F2 males
Relative weight
Fl males
F2 males
Absolute weight
Fl females
F2 females
Relative weight
Fl females
F2 females
Spleen weight (percent
mg/kg-day (M)
Absolute weight
2 weeks
6 weeks
14 weeks
Relative weight
2 weeks
6 weeks
14 weeks
mg/kg-day (F)
Absolute weight
2 weeks
6 weeks
(percent
0
0
0
0
0
0
0
0
0
change c
0
0
0
0
0
0
0
0
0
0
Results
change compared to c
50
0
4
1
4
8
8
6
6
ompared to control)
151
-6
-7
-10
0
-5
-5
171
0
0
ontrol)
250
-1
1
-3
0
-5
4
-4
1
381
-2
-10
-7*
0
-5
0
422
0
5
750
-29*
-27*
-12*
-18*
-34*
-26*
-14*
-17*
960
-19
-24*
-4
-14*
-14*
5
1,069
-8
0
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Asoetal. (2005)
Rat (Crj:CD(SD)IGS);
24 sex/generation/group; assessed
in male offspring/litter in Fl and F2
offspring
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
FO and Fl exposed for 4 weeks prior
to mating, through mating for
10 weeks, and until weaning of
offspring (females) or necropsy
(males)
Nagao et al. (2000)
Rat (Sprague-Dawley);
25 sex/generation/group
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
Results
14 weeks
0
0
-5
0
Relative weight
2 weeks
6 weeks
14 weeks
0
0
0
-3
-4
0
0
0
-5
0
8
5
Spleen weight (percent change compared to control)
mg/kg-day
Absolute weight,
Fl males
F2 males
0
PND21
0
0
100
1
-12
200
3
-8
400
-16*
-25*
Relative weight, PND21
Fl males
F2 males
Absolute weight,
FO males
Fl males
FOfemales
Fl females
0
0
study termination
0
0
0
0
-2
-9
-7
-6
-2
2
4
-8
-5
-7
6
2
-13*
-20*
-4
-8
-2
-2
Relative weight, study termination
FO males
Fl males
FOfemales
Fl females
0
0
0
0
-7
-7
-6
-6
-7
-7
0
-6
0
-7
-6
-6
Spleen weight (percent change compared to control)
mg/kg-day
0
20
100
500
Absolute weight
FO males
Fl males
FOfemales
Fl females
0
0
0
0
8
-6
-1
-4
4
-3
1
6
-1
-12*
-1
1
Relative weight
FO males
Fl males
0
0
14
0
7
7
7
0
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
FO males and females: Exposure for
12 weeks prior to mating, 2 weeks
cohabitation, and until necropsy at
23 weeks of age (males) or until
postpartum day 22 (females); Fl
animals were exposed from weaning
until necropsy at PND 22
Thymus weights0
NTP (1997b)
Rat (F344); 15 males/group
0, 300, 2,800, 25,000 ppm
0, 20, 200, 2,200 mg/kg-dayb
Diet
10 weeks
NTP (1997b)
Rat (F344); 15 males/group
0, 300, 900, 2,800, 8,300,
25,000 ppm
0, 30, 60, 180, 550, "high"
mg/kg-dayb
Diet
26 weeks
Tvl et al. (2004)
Rat (CD); 30 FO and Fl parental
rats/sex/group; assessed in
54-86 male offspring/group and
43-87 female offspring/group
(>3 sex/litter/group if possible)
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
Exposure 10 weeks prior to mating
and through mating, gestation, and
lactation (females); or for 21 days
after mating (males).
Results
FOfemales 005
Fl females 0
-5 5
Thymus weight (percent change compared to control)
mg/kg-day 0 20 200
absolute weight 06-2
relative weight 060
Thymus weight (percent change compared to control)
mg/kg-day 0 30 60 180
absolute weight 0 5 52 3
relative weight 0 1 46 6
Thymus weight (percent change compared to control)
mg/kg-day 0 50 250
Absolute weight
Flmale 071
F '2 male 0 -3 -5
Fl female 02-3
F2 female 0 -3 -5
Relative weight
Fl male 060
F '2 male 0 -2 -6
Fl female 00-2
F2 female 0 -5 -8
0
5
2,200
-14
23*
550 ND
12 -27
12 10
750
-17*
-14*
-22*
-15*
2
-2
-2
-4
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Nagao et al. (2000)
Rat (Sprague-Dawley);
25 sex/generation/group
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
FO males and females: Exposure for
12 weeks prior to mating, 2 weeks
cohabitation, and until necropsy at
23 weeks of age (males) or PND 22
(females); Fl animals: Exposure
from weaning until necropsy at
PND 22
Asoetal. (2005)
Crj:CD(SD)IG rats,
24 rats/sex/generation/group;
assessed in 1 male/litter in Fl and F2
offspring
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
FO and Fl exposed for 4 weeks prior
to mating, through mating for
10 weeks, and until weaning of
offspring (females) or necropsy
(males).
Results
Note: Thymus weights were not recorded for FO or Fl parental animals.
There were no significant effects on relative thymus weights of Fl or F2
offspring.
Thymus weight (percent change compared to control)
mg/kg-day 0 20 100 500
Absolute weight
FO males 0 -12 -15 -10
Fl males 0 -4 -18 -12
FOfemales 0 -10 2 -6
Fl females 0 5 -13 13
Relative weight
FO males 0 -9 -14 -3
Fl males 0 0 -12 0
FOfemales 0-9 2-7
Fl females 0 5 -14 13
Thymus weight, PND 21 (percent change compared to control)
mg/kg-day 0 100 200 400
Absolute weight
Flmale 0656
F '2 male 0-1 3-8
Relative weight
Flmale 0459
F '2 male 042-2
Other changes
Piersma et al. (2000)
Rat (Harlan Cpb-WU);
10 females/group
0, 270, 350, 450, 580, 750, 970,
1,250, 1,600, 2,100 mg/kg-day
Gavage
CDs 6-15 or 6-20
Histopathological effects in the spleen at GD 21
Dose related increase in the extent of extramedullary hematopoiesis (data
presented graphically). The severity of the effect was reportedly
increased. The effect was classified as normal (0), minimal (1), slight (2),
moderate (3), marked (4), or severe (5). It was also noted in the study
report that, "pregnant controls showed elevated extramedullary
hematopoiesis compared to nonpregnant females (quantitative data not
reported), which was further increased after exposure in all dose groups."
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
BIBRA (1978)
Rat (Wistar); 27/sex/group or
45/sex/group (control); interim
sacrifices of 9 controls/sex/group
and 6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
0, 151, 381, 960 mg/kg-day (males)";
0, 171, 422, 1,069 mg/kg-day
(females)b
Diet
14 weeks
NTP (1997b)
Rat (F344); 15 males/group
0, 300, 2,800, 25,000 ppm
0, 20, 200, 2,200 mg/kg-dayb
Diet
10 weeks
NTP (1997b)
Rat (F344); 15 males/group
0, 300, 900, 2,800, 8,300,
25,000 ppm
0, 30, 60, 180, 550, "high"
mg/kg-dayb
Diet
26 weeks
Results
Percent incidence
mg/kg-day (M) 0 151
381 960
Hemorrhage in medulla/congested; thymus
6 weeks 22 NE
14 weeks 0 NE
NE 0
NE 0
Atrophy of medullas; thymus
14 weeks 0 NE
NE 0
Hemorrhage/congested; lymph nodes
14 weeks 7 NE
mg/kg-day (F) 0 171
NE 7
422 1,069
Hemorrhage in medulla/congested; thymus
6 weeks 22 NE
14 weeks 4 NE
NE 0
NE 7
Atrophy of medulla; thymus
14 weeks 4 NE
NE 0
Hemorrhage/congested; lymph nodes
14 weeks 4 NE NE 7
Note: It is unclear based on the study report if the spleen was examined
histopathologically. However, no effects were reported.
Spleen and thymus were examined histopatholo
effects reported in control or high-dose animals
reported).
Spleen and thymus were examined histopatholo
effects reported in control or high-dose animals
reported).
gically. No significant
(quantitative data not
gically. No significant
(quantitative data not
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1997b)
Rat (F344), 60/sex/group; assessed
in 10 rats/sex/group at 15-month
interim sacrifice and 50
rats/sex/group at study termination
0, 3,000, 6,000, 12,000 ppm (males);
0, 6,000, 12,000, 24,000 ppm
(females)
0, 120, 240, 500 mg/kg-day (males)";
0 300, 600, 1,200 mg/kg-day
(females)"
Diet
2 years
Results
Percent incidence
mg/kg-day (M) 0 120
240 500
15 months
lymph node: deep 50 NE
cervical; hemorrhage
lymph node: 100 NE
mediastinal;
hemorrhage
lymph node: 20 0
mandibular;
hemorrhage
lymph node: 0 0
mesenteric;
hemorrhage
spleen: hematopoietic 20 0
cell proliferation
spleen: pigmentation; 100 100
hemosiderin
0 0
100 100
10 0
20 0
0 10
100 70
2 years
bone marrow: 2 8
hypercellularity
lymph node: iliac; 4 0
hemorrhage
lymph node: 22 23
mediastinal;
hemorrhage
lymph node: 0 5
pancreatic;
hemorrhage
lymph node: 2 4
mandiubular;
congestion
lymph node: 8 6
mandibular;
hemorrhage
lymph node: 2 6
mesenteric;
hemorrhage
spleen: hematopoietic 4 16
cell proliferation
2 0
0 0
0 0
0 5
0 0
4 12
0 8
10 14
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
spleen: pigmentation; 28 2
hemosiderin
thymus: hemorrhage 2 2
mg/kg-day (F) 0 300
4 12
0 0
600 1,200
15 months
lymph node: NE 100
mediastinal;
hemorrhage
lymph node: 0 0
mandibular;
hemorrhage
spleen: hematopoietic 10 30
cell proliferation
spleen: pigmentation; 100 100
hemosiderin
thymus: hemorrhage 0 11
100 100
0 10
30 30
100 100
0 0
2 years
bone marrow: 2 4
hypercellularity
lymph node: 8 9
mediastinal;
hemorrhage
lymph node: 0 0
pancreatic;
hemorrhage
lymph node: renal; 0 18
hemorrhage
lymph node: 10 18
mandibular;
hemorrhage
lymph node: 6 6
mesenteric;
hemorrhage
spleen: hematopoietic 20 24
cell proliferation
spleen: pigmentation; 38 40
hemosiderin
thymus: hemorrhage 2 0
0 2
6 7
6 0
13 0
6 10
6 4
16 28
42 58
2 2
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
Asoetal. (2005)
Crj:CD(SD)IG rats,
24 rats/sex/generation/group;
assessed in 1 male/litter in Fl and
F2 offspring
0,100, 200, 400 mg/kg-day
Gavage
Multigenerational study
FO and Fl exposed for 4 weeks prior
to mating, through mating for
10 weeks, and until weaning of
offspring (females) or necropsy
(males).
Spleen and thymus were examined histopathologically. No significant
treatment-related effects observed in FO or Fl parental males or females
(data not provided).
Monsanto (1983)
Percent incidence
Rat (Sprague-Dawley);
25/sex/group; interim sacrifice of
10 rats/sex/group at 7 weeks
0, 51, 218, 789 mg/m3
Inhalation (whole-body)
13 weeks
mg/kg-day
Males
sinusoidal congestion,
6 weeks
yellow-brown pigment,
13 weeks
10
Females
sinusoidal congestion,
6 weeks
yellow-brown pigment,
13 weeks
20
51
218
789
NTP U997a)
Percent incidence
Rat (F344); 50-60/sex/group;
interim sacrifice of 10
rats/sex/group at 15 months
0,12,000 ppm (males); 0,
24,000 ppm (females)
0, 500 mg/kg-day (males)b; 0,
1,200 mg/kg-day (females)b
Diet
4 exposure protocols: ad libitum
feeding, weight-matched controls,
restricted feed (2 years), and
restricted feed (lifetime)
Diet
Ad libitum and weight matched
mg/kg-day (M)
0(ad libitum)
0 (weight-
matched)
500
lymph node: deep
cervical; hemorrhage,
15 months
lymph node:
mediastinal;
hemorrhage,
15 months
50
100
20
NE
NE
20
100
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
2 years to lifetime
Results
lymph node:
mandibular;
hemorrhage,
15 months
lymph node: 0
mesenteric;
hemorrhage,
15 months
spleen: hematopoietic 20
cell proliferation,
15 months
spleen: pigmentation; 100
hemosiderin,
15 months
bone marrow: 2
hypercellularity, 2 years
lymph node: iliac; 4
hemorrhage, 2 years
lymph node: 22
mediastinal;
hemorrhage, 2 years
lymph node: 0
pancreatic;
hemorrhage, 2 years
lymph node: 2
mandiubular;
congestion, 2 years
lymph node: 8
mandibular;
hemorrhage, 2 years
lymph node: 2
mesenteric;
hemorrhage, 2 years
spleen: hematopoietic 4
cell proliferation,
2 years
spleen: pigmentation; 28
hemosiderin, 2 years
thymus: hemorrhage, 2
2 years
mg/kg-day (F) 0 (ad libitum)
0
0
90
4
0
29
0
0
17
6
12
4
0
0 (weight-
matched)
0
10
70
0
0
0
5
0
12
8
14
12
0
1,200
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
lymph node: NE
mediastinal;
hemorrhage,
15 months
lymph node: 0
mandibular;
hemorrhage,
15 months
spleen: hematopoietic 10
cell proliferation,
15 months
spleen: pigmentation; 100
hemosiderin,
15 months
bone marrow: 2
hypercellularity, 2 years
lymph node: 8
mediastinal;
hemorrhage, 2 years
lymph node: renal; 0
hemorrhage, 2 years
lymph node: 10
mandibular;
hemorrhage, 2 years
lymph node: 6
mesenteric;
hemorrhage, 2 years
spleen: hematopoietic 20
cell proliferation,
2 years
spleen: pigmentation; 38
hemosiderin, 2 years
thymus: hemorrhage, 2
2 years
Feed-restricted, 2 years or lifetime
NE
10
100
23
16
26
100
10
30
100
10
28
58
mg/kg-day (M)
0 500 500
(2 years) (2 years) 0 (lifetime) (lifetime)
lymph node:
mediastinal;
hemorrhage,
15 months
100
NE
NA
NA
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
lymph node: 10 10
mandibular;
hemorrhage,
15 months
spleen: pigmentation; 0 10
hemosiderin,
15 months
thymus: hemorrhage, 10 10
15 months
bone marrow: 4 4
hypercellularity, 2 years
lymph node: deep 0 0
cervical; hemorrhage,
2 years
lymph node: iliac; 0 0
hemorrhage, 2 years
lymph node: 8 11
mediastinal;
hemorrhage, 2 years
lymph node: 0 5
pancreatic;
hemorrhage, 2 years
lymph node: 10 15
mandibular;
hemorrhage, 2 years
lymph node: 4 2
mesenteric;
hemorrhage, 2 years
spleen: hematopoietic 10 8
cell proliferation,
2 years
spleen: pigmentation; 12 8
hemosiderin, 2 years
0 1,200
mg/kg-day (F) (2 years) (2 years)
lymph node: 100 0
mediastinal;
hemorrhage,
15 months
lymph node: 0 30
mandibular;
hemorrhage,
15 months
NA NA
NA NA
NA NA
2 4
4 0
0 4
0 9
4 0
12 10
0 4
16 8
16 6
0 1,200
(lifetime) (lifetime)
NA NA
NA NA
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1982)
F344 rats; 50/sex/group
0, 6,000, 12,000 ppm
0, 474, 947 mg/kg-day (males)"; 0,
550, 1,100 mg/kg-day (females)"
Diet
Results
lymph node:
mesenteric;
hemorrhage,
15 months
spleen: hematopoietic
cell proliferation,
15 months
spleen: pigmentation;
hemosiderin,
15 months
bone marrow:
hypercellularity, 2 years
lymph node:
mediastinal;
hemorrhage, 2 years
lymph node: renal;
hemorrhage, 2 years
lymph node:
mandibular;
hemorrhage, 2 years
lymph node:
mesenteric;
hemorrhage, 2 years
spleen: hematopoietic
cell proliferation,
2 years
spleen: pigmentation;
hemosiderin, 2 years
thy m us: atrophy,
2 years
thymus: hemorrhage,
2 years
0 10 NA NA
30 10 NA NA
90 90 NA NA
0242
10 13 5 6
10 13 0 0
10 14 12 14
4444
26 16 26 16
42 36 36 48
0200
0020
Percent incidence
mg/kg-day (F)
bone marrow.
hypoplasia
spleen: pigmentation;
NOS
0 550 1,100
040
020
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
28 weeks (males) or 103 weeks
(females)
NTP (1982)
B6C3Fi mice; 50/sex/group
0, 6,000, 12,000 ppm
0, 474, 947 mg/kg-day (males)"; 0,
550, 1,100 mg/kg-day (females)"
Diet
103 weeks
BIBRA (1978)
Rat (Wistar); 27/sex/group or
45/sex/group (control); interim
sacrifices of 9 controls/sex/group
and 6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
0, 151, 381, 960 mg/kg-day (males)"
0, 171, 422, 1,069 mg/kg-day
(females)"
Results
spleen: hemosiderosis 6 4
lymph node: 0 0
mediastinal;
hemorrhage
lymph node: 0 0
pancreatic;
hemorrhage
lymph node: 0 0
mesenteric;
hemorrhage
thymus: hemorrhage 0 4
thy m us: atrophy 0 4
Note: Males were not examined histopathologically.
8
2
2
4
4
7
Percent incidence
mg/kg-day (M) 0 474
bone marrow: 0 2
hyperplasia;
hematopoietic
lymph node: 0 4
mesenteric;
hemorrhage
thymus: atrophy 0 0
mg/kg-day (F) 0 550
bone marrow: 6 2
hyperplasia;
hematopoietic
spleen: congestion; nos 0 0
spleen: hyperplasia; 4 2
hematopoietic
thymus: atrophy 5 0
947
0
14
5
1,100
2
2
0
0
Percent change compared to control
mg/kg-day (F) 0 171 422
hemoglobin 002
concentration
hematocrit (packed cell 002
volume)
spleen histopathology No lesions were noted in the spleen
mg/kg-day (M) 0 151 381
1,069
4
0
960
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Diet
14 weeks
Mononuclear cell leukemia
NTP (1982)
B6C3Fi mice; 50/sex/group
0, 6,000, 12,000 ppm
0, 474, 947 mg/kg-day (males)"; 0,
550, 1,100 mg/kg-day (females)"
Diet
103 weeks
NTP (1982)
F344 rats; 50/sex/group
0, 6,000, 12,000 ppm
0, 474, 947 mg/kg-day (males)"; 0,
550, 1,100 mg/kg-day (females)"
Diet
28 weeks (males) or 103 weeks
(females)
NTP (1997b)
F344 rats; 60/sex/group; assessed in
10 rats/sex/group at 15-month
interim sacrifice and
50 rats/sex/group at study
termination
0, 3,000, 6,000, 12,000 ppm (males);
0, 6,000, 12,000, 24,000 ppm
(females)
0, 120, 240, 500 mg/kg-day (males)";
0 300, 600, 1,200 mg/kg-day
(females)"
Diet
2 years
Results
hemoglobin 022
concentration
hematocrit (packed cell 000
volume)
spleen histopathology No lesions were noted in the
Percent incidence
mg/kg-day (M) 0 474
leukemia, multiple 2 0
organs
mg/kg-day (F) 0 550
leukemia, liver 0 0
Percent incidence
mg/kg-day (M) 0 474
mononuclear cell NE NE
leukemia
mg/kg-day (F) 0 550
mononuclear cell 14 14
leukemia, total
multiple organs 12 12
spleen 0 2
liver 2 0
Percent incidence at study termination
mg/kg-day (M) 0 120 240
mononuclear cell 62 56 68
leukemia
mg/kg-day (F) 0 300 600
mononuclear cell 42 40 42
leukemia
-6
_9*
spleen
947
0
1,100
2
947
NE
1,100
36
34
2
0
500
60
1,200
38
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1997a)
F344 rats; 50-60/sex/group; interim
sacrifice of 10 rats/sex/group at
15 months
0, 12,000 ppm (males); 0,
24,000 ppm (females)
0, 500 mg/kg-day (males)b; 0,
1,200 mg/kg-day (females)b
Diet
4 exposure protocols: ad libitum
feeding, weight-matched controls,
restricted feed (2 years), and
restricted feed (lifetime)
2 years to lifetime
Results
Percent incidence at study termination
Ad libitum and weight-matched
mg/kg-day (M) 0 0
(ad libitum) (weight-
matched)
mononudear cell 62 30
leukemia
mg/kg-day (F) 0 0
(ad libitum) (weight-
matched)
mononudear cell 42 26
leukemia
500
60
1,200
38
Feed restricted 2-year exposure
mg/kg-day (M) 0
mononudear cell 42
leukemia
mg/kg-day (F) 0
mononudear cell 32
leukemia
500
54
1,200
36
Feed restricted lifetime exposure
mg/kg-day (M) 0
mononudear cell 78
leukemia
mg/kg-day (F) 0
mononudear cell 58
leukemia
500
72
1,200
78
1
2
3
4
5
6
7
*Statistically significant (p <0.05) relative to controls based on statistics performed by the study authors.
aPercent change compared to control calculated as 100 x ([treated value - control value] 4- control value).
bCalculated as follows: [% in diet x intake food/water (mg)] -f body weight (kg) = mg/kg-day.
NE = not examined; NOS = not otherwise specified
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
i Spleen Relative Weight
J. Spleen Absolute Weight
T Thymus Relative Weight
T Thymus Absolute Weight
BIBRA, 1978; 14wks; male Wistar rats
BIBRA, 1978; 14wks; female Wistar rats
Aso etal., 2005; 2-gen; study termination; FO, Fl
male and female S-D rats
Aso etal., 2005; 2-gen; PND 21; Fl, F2 male S-D
rats
Tyl et al., 2004; 2-gen; PND21; Fl, F2 male and
female CD rats
BIBRA, 1978; 14wks; female Wistar rats
BIBRA, 1978; 14wks; male Wistar rats
Aso etal., 2005; 2-gen; study termination; FO, Fl
male and female S-D rats
Aso etal, 2005; 2-gen; PND 21; Fl, F2 male S-D
rats
Nagaoetal, 2000; 2-gen; FO males & Fl, FO
female S-D rats
Nagaoetal., 2000; 2-gen; Fl male S-D rats
Tyl et al, 2004; 2-gen; PND21; Fl, F2 male and
fern ale CD rats
NTP 1997b; lOwks; male F344 rats
NTP 1997b; 26wks; male F344 rats
Aso etal, 2005; 2-gen; PND 21; Fl, F2 male S-D
rats
Nagao et al, 2000; 2-gen; male & female S-D rats
Tyl et al, 2004; 2-gen; PND21; Fl, F2 male and
fern ale CD rats
NTP 1997b; lOwks; male F344 rats
NTP 1997b; 26wks; male F344 rats
Aso etal, 2005; 2-gen; PND 21; Fl, F2 male S-D
rats
Nagaoetal, 2000; 2-gen; male & female S-D rats
Tyl et al, 2004; 2-gen; PND21; Fl, F2 male and
female CD rats
o — a — a
a — a — •
Q — B — Q
Q — B — •
a a a
a a D
• = statistically significant
D = not statistically significant
10
100
1000
10000
Dose (mg/kg-day)
2
3
Figure 3-18. Exposure-response array of hematopoietic effects following oral
exposure to BBP: spleen and thymus weights.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
o
13
o
3
O
3
C
t—
n>
C
NTP 1997b; lOwks; spleen/thymus
male F344 rats
B1BRA, 1978; 14wks; thyrn us/lymph node
male Wistar rats
B1BRA, 1978; 14wks; thymus/lymph node
female Wistar rats
NTP 1997b; 26wks; spleen/thymus
male F344 rats
Aso et al., 2005; 2-gen; spleen/thymus
FO, Fl parental S-D rats
NTP 1997a; 15 mo. & 2 yr; spleen/lymph
node/bone marrow
male F344rats
NTP 1997a; 15 mo. & 2 yr; spleen/lymph
node/bone marrow
female F344rats
NTP 1982; 103wks; female F344 rats
NTP 1982; 103wks; male B6C3F1 mice
NTP 1982; 103wks; female B6C3F1 mice
BIBRA, 1978; 14wks; male
NTP 1982; 103wks; female F344 rats
NTP 1997b; 2 yr; male F344 rats
NTP 1997b; 2 yr; female F344 rats
NTP 1997a; 2 yr and study termination; male
F3 44 rats
NTP 1997a; 2 yr and study termination; female
F3 44 rats
Q—B B B
Q—B—B
O—B
D B
B—B
B B
• - statistically significant
D - not statistically significant
10 100 1000
Dose (mg/kg-day)
10000
2
3
Figure 3-19. Exposure-response array of hematopoietic histopathological
effects following oral exposure to BBP.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.3.8. Thyroid Effects
2
3
Table 3-30. Evidence pertaining to thyroid effects in animals following oral
exposure to BBP
Reference and study design
Results3
Thyroid weight
Asoetal. (2005)
Rat(Crj:CD(SD)IGS);
24 rats/sex/generation/group
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
FO and Fl exposed for 4 weeks prior
to mating, through mating for
10 weeks, and until weaning of
offspring (females) or necropsy
(males)
Nagao et al. (2000)
Rat (Sprague-Dawley);
20-25 rats/sex/generation/group
0, 20, 100, 500 mg/kg-day
Diet
Multigenerational study
FO males and females: Exposure for
12 weeks prior to mating, 2 weeks
cohabitation, and until necropsy at
23 weeks of age (males) or PND 22
(females); Fl animals: Exposure
from weaning until necropsy at
PND 22
Thyroid weight (percent
mg/kg-day
change
0
compared to control)
100
200
400
Absolute weight
FO males
Fl males
FO females
Fl females
0
0
0
0
-6
0
-4
5
5
3
-6
-2
9
21
10
-1
Relative weight
FO males
Fl males
FOfemales
Fl females
Thyroid weight (percent
mg/kg-day
0
0
0
0
change
0
-10
0
-4
0
compared to control)
20
0
2
-12
-8
100
10
24*
9
-3
500
Absolute weight
FO males
Fl males
FOfemales
Fl females
0
0
0
0
2
3
12
-9
7
5
1
-2
0
7
8
4
Relative weight
FO males
Fl males
FOfemales
Fl females
0
0
0
0
3
-3
14
-10
6
3
0
-2
9
11*
7
4
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
BIBRA (1978)
Rat (Wistar); 27/sex/group or
45/sex/group (control); interim
sacrifices of 9 controls/sex/group
and 6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
0, 151, 381, 960 mg/kg-day (males)"
0, 171, 422, 1,069 mg/kg-day
(females)"
Diet
14 weeks
Tvl et al. (2004)
Rat (CD); 30 FO and Fl parental
rats/sex/group
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
Exposure 10 weeks prior to mating
and through mating, gestation, and
lactation (females) or through
21 days after end of mating (males)
Results3
Thyroid weight (percent
mg/kg-day (M)
change compared to control)
0 151
381 960
Absolute weight
2 weeks
6 weeks
14 weeks
0 6
0 20*
0 4
8 4
-18 -13
3 4
Relative weight
2 weeks
6 weeks
14 weeks
mg/kg-day (F)
0 10
0 26*
0 14
0 171
8 10
-11 -2
14 17
422 1,069
Absolute weight
2 weeks
6 weeks
14 weeks
0 19
0 -18*
0 -1
29* 35*
-15 -13
-3 1
Relative weight
2 weeks
6 weeks
14 weeks
0 10
0 -19*
0 -2
28* 43*
-17 -8
-2 5
No significant treatment-related effects on absolute or relative thyroid
weight were reported in FO or Fl parental males or females (data not
provided).
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results3
Thyroid hormones
Nagao et al. (2000)
Rat (Sprague-Dawley);
20-25 parental
rats/sex/generation/group;
37-48 Fl offspring/group (from
18-24 litters/group)
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
FO males and females: Exposure for
12 weeks prior to mating, 2 weeks
cohabitation, and until necropsy at
23 weeks of age (males) or PND 22
(females) Fl animals: Exposure from
weaning until necropsy at PND 22
NTP (1997b)
Rat (F344); 60/sex/group; assessed
in 10 rats/sex/group at 6, 8, and/or
15 months and at study termination
0, 3,000, 6,000, 12,000 ppm (males);
0, 6,000, 12,000, 24,000 ppm
(females)
0, 120, 240, 500 mg/kg-day
(males)"; 0 300, 600,
1,200 mg/kg-day (females)b
Diet
Percent change compared to
parental animals) or PND 22
mg/kg-day (M)
TSH FO, parental
T3 FO, parental
T4 FO, parental
TSH Fl, parental
T3 Fl, parental
T4 Fl, parental
TSH Fl, weanling
T3 Fl, weanling
T4 Fl, weanling
mg/kg-day (F)
TSH FO, parental
T3 FO, parental
T4 FO, parental
TSH Fl, parental
T3 Fl, parental
T4fl, parental
TSH Fl, weanling
T3 Fl, weanling
T4 Fl, weanling
control at study termination (FO and
Fl
(Fl weanling rats)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
20
-9
0
0
12
14
10
-1
23*
4
20
12*
13
-5
-5
0
12
3
0
4
Note: TSH, T4, and T3 for all other lifestages examined
100
-12*
0
-4
1
14
-1
-15*
8
2
100
1
13
-16
-7
0
8
0
-17*
4
500
-10
-11*
-21*
5
14
-21*
-19*
-8
2
500
6
13
-21*
-10
0
10
9
-33*
9
were not affected.
Percent change
mg/kg-day (M)
0
120
240
500
TSH
6 months
8 months
15 months
study termination
0
0
0
0
167
200*
-50
100
100
inn*
-50
33
100
0
T3
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
2 years
Thyroid histopathology
NTP (1997b)
Rat (F344); 60/sex/group; interim
sacrifice of 10 rats/sex/group at
5 months
0, 3,000, 6,000, 12,000 ppm (males);
0, 6,000, 12,000, 24,000 ppm
(females)
0, 120, 240, 500 mg/kg-day
(males)b; 0 300, 600,
1,200 mg/kg-day (females)b
Diet
2 years
6 months
15 months
study termination
T4
6 months
15 months
study termination
mg/kg-day (F)
TSH
6 months
15 months
study termination
T3
6 months
15 months
study termination
T4
6 months
15 months
study termination
Percent incidence
mg/kg-day (M)
Ultimobranchial cyst
15 months
2 years
C-cell, hyperplasia
15 months
2 years
Follicle, cyst
15 months
2 years
0
0
0
0
0
0
0
-
-
-
0
4
0
8
0
4
Results3
25
-1
-32
0
0
-25
300
-50
0
0
15
-1
-22
0
0
0
0 120
0
4
0
20
0
4
3
7
3
0
25
0
600
0
100
0
-5
-7
-25
0
0
0
240
10
2
10
24
10
0
-4
1
0
0
-25
0
1,200
-100
100
0
-30*
-22*
-36*
-25
-33*
0
500
20
4
0
14
10
8
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results3
mg/kg-day (F)
300
600
1,200
Ultimobranchial cyst
15 months
2 years
10
4
30
2
0
2
0
0
C-cell, hyperplasia
15 months
2 years
0
12
10
12
10
14
0
6
Follicle, cyst
2 years
Follicular cell, hyperplasia
2 years
0
0
0
NTP U997b)
Rat (F344); 15 males/group
0, 300, 2,800, 25,000 ppm
0, 20, 200, 2,200 mg/kg-dayb
Diet
10 weeks
No significant effects on thyroid histopathology reported in control or high-
dose animals (quantitative data not reported).
NTP U997b)
Rat (F344); 15 males/group
0, 300, 900, 2,800, 8,300,
25,000 ppm
0, 30, 60, 180, 550, "high"
mg/kg-dayb
Diet
26 weeks
No significant effects on thyroid histopathology reported in control or high-
dose animals (quantitative data not reported).
Asoetal. (2005)
Rat(Crj:CD(SD)IGS);
24 rats/sex/generation/group
0,100, 200, 400 mg/kg-day
Gavage
Multigenerational study
FO and Fl exposed for 4 weeks prior
to mating, through mating for
10 weeks, and until weaning of
offspring (females) or necropsy
(males)
No significant treatment-related effects on thyroid histopathology were
reported by the study authors in FO or Fl parental animals.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results3
Tvl et al. (2004)
Rat (CD); 30 FO and Fl parental
rats/sex/group
0, 750, 3,750,11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
Exposure 10 weeks prior to mating
and through mating, gestation, and
lactation (females) or through
21 days after end of mating (males)
No significant treatment-related effects on thyroid histopathology were
reported by the study authors in control or high-dose parental males or
females (data not provided).
BIBRA (1978)
Rat (Wistar); 27/sex/group or
45/sex/group (control); interim
sacrifices of 9 controls/sex/group
and 6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
0,151, 381, 960 mg/kg-day (males)"
0,171, 422,1,069 mg/kg-day
(females)"
Diet
14 weeks
No significant treatment-related effects on thyroid histopathology were
reported by the study authors for males or females.
1
2
3
4
5
*Statistically significant (p <0.05) relative to controls based on statistics performed by the study authors.
aPercent change compared to control calculated as 100 x ((treated value - control value) -f control value).
"Calculated as follows: [% in diet x intake food/water (mg)] 4- body weight (kg) = mg/kg-day.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
BIBRA, 1978; 14wks; male Wistar rats
B1BRA, 1978; 14wks; female Wistar rats
Asoetal., 2005; 2-gen; FO male&FO, Fl female S-D rats
Asoetal, 2005; 2-gen; PI male S-D rats
Nagao et al, 2000; 2-gen; FO males & FO, Fl female S-D rats
Nagaoetal., 2000; 2-gen; Fl male S-D rats
Tyl et al., 2004; 2-gen; FO, Fl parental CD rats
BIBRA, 1978; 14wks; male Wistar rats
BIBRA, 1978; 14wks; female Wistar rats
Aso etal, 2005; 2-gen; FO, Fl male & female S-D rats
Nagao et al, 2000; 2-gen; FO, Fl male & female S-D rats
Tyl et al, 2004; 2-gen; FO, Fl parental CD rats
NTP, 1997b; 2 yr; TSH, T3, T4 male F344 rats
NTP, 1997b; 2 yr; TSH, T4 female F344 rats
NTP, 1997h; 2 yr; T3 female F344 rats
Nagao et al, 2000; 2-gen; T3, T4 FO parental male S-D rats
Nagao et al, 2000; 2-gen; T4 FO parental female S-D rats
Nagaoetal, 2000; 2-gen; TSH Fl weanling male, T3 Fl
weanling female S-D rats
H
f
NTP, 1997b; lOwks; male F344 rats
BIBRA, 1978; 14wks; female Wistar rats
BIBRA, 1978; 14wks; male Wistar rats
NTP, 1997b; 26wks; male F344 rats
Aso etal, 2005; 2-gen; FO, Fl male & female S-D rats
Tyl et al, 2004; 2-gen; FO, Fl parental CD rats
NTP 1997b; 15 mo or 2 yr; male F344 rats
NTP 1997b; 15 mo or 2 yr; female F344 rats
Q B B
B B B
B
-B B
B B B
B B B
D
= statistically significant
= not statistically significant
10
100
1000
10000
Dose (mg/kg-day)
2
3
Figure 3-20. Exposure-response array of thyroid effects following oral
exposure to BBP.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.3.9. Immune Effects
2 Table 3-31. Evidence pertaining to immune effects in animals following oral
3 exposure to BBP
Reference and study design
BIBRA (1978)
Rat (Wistar); 27/sex/group or
45/sex/group (control); interim
sacrifices of 9 controls/sex/group
and 6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
0, 151, 381, 960 mg/kg-day (males)
0, 171, 422, 1,069 mg/kg-day
(females)
Diet
14 weeks
Butala et al. (2004)
B6C3Fi mice, 10 female mice/dose
0, 100% BBP
10 applications of 50 nL BBP over
2 weeks then challenged with BBP
7 days later. Animals sacrificed
7 days after challenge.
Dearman et al. (2009)
BALB/c mice, 10 mice/dose
Results
Leukocyte count (percent change compared to control)
mg/kg-day 0 151 381
male 0 -10 -10
mg/kg-day 0 171 422
female 0 -16 -5
960
-4
1,069
-8
Percent change compared to control
Doses (%) 0
100
serum IgE 0 6.1
IL-4fromConA- 0 -52
stimulated lymph node
cells
IL-13fromConA- 0 36
stimulated lymph node
cells
IL-4m-RNAfrom 0 266
stimulated lymph nodes
IL-4 m-RNAfrom Not able to be determined due to vehicle control
stimulated lymph nodes values being indistinguishable from background
Percent change compared to control
Doses (%) 0 5 10 50
100
Antibodies
IgE : No effect (data not reported)
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results
0, 5, 10, 50, or 100% BBP
Dermal
15 applications of 100 ul at site of
ovalbumin injection (21-day
treatment period consisting of 5
consecutive days of treatment
followed by 2 days of rest)
Ovalbumin s.c. injections: initial
injection of 1 u.g on study day 0;
follow-up injections of 0.1 u.g on
study day 10 and 15
IgG:
-9
12
-9
29
1
2
3
Con A = Concanavalin A; IL = interleukin; m-RNA = messenger ribonucleic acid
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.3,10. Neurological Effects
2
3
Table 3-32. Evidence pertaining to neurological effects in animals following
oral exposure to BBP
Reference and study design
Zhuang et al. (2008)
Rat (Wistar); 20 females/dose;
neurobehavioral development was
assessed in 15-45 pups/sex/dose
0, 0.05, 0.25, 0.75%
0, 50, 250, 750 mg/kg-day (dams)
Diet
Dams: PNW 4 through mating,
gestation, and lactation; Fl pups:
GDO-PNW6
Betz et al. (2013)
Rat (Sprague-Dawley); male (NR)
0, 5, 10 ppm (equivalent to 0, 2,
4 mg/kg-day as indicated by study
authors)
Drinking water
Daily for 15 weeks (PND 40-140)
Results
Neurobehavioral effects in Fl offspring
In Fl male rats, the following changes were observed: statistically
significantly impaired cliff avoidance on PND 7 in high-dose rats (score of
41 vs 61 points in controls) and statistically significantly depressed air
righting on PND 14 in all exposure groups (~30-40% lower than controls3
based on visual inspection of data shown graphically). Other statistically
significant neurobehavioral differences were also reported in male
offspring, including delayed surface righting on PND 3 (mid-dose group
only), increased locomotion in the open field test (low and high doses)
and delayed escape latency in the Morris water maze test (low dose
only, on 5th day only). No significant neurobehavioral effects were
observed in Fl females.
Fear (freezing) during cue phase of fear conditioning
Significant decreased expression of fear during Cue phase with 10 ppm
exposure.
Fear (freezing) during inter-trial interval phase of fear conditioning
Significant decrease in expression of fear during inter-trial interval phase
with 10 ppm exposure.
Open field activity in inner portion
No significant change.
Open field activity in outer portion
No significant change.
Open field grooming activity
No significant change.
Open field rearing activity
No significant change.
Social test (contact behavior)
No significant change.
Social test (non-contact behavior)
Increased sniffing and approaching (non-contact behavior) with both
treatment.
Social test (self-directed behavior)
No significant change.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.3.11. Other Toxicity Effects
2
3
Table 3-33. Evidence pertaining to other toxicity effects in animals following
oral exposure to BBP
Reference and study design
Results
Body-weight effects"
BIBRA (1978)
Rat (Wistar)
27/sex/group or 45/sex/group
(control); interim sacrifice of
9 controls/sex/group and 6 treated
rats/sex/group at 2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
0, 151, 381, 960 mg/kg-day
(males)"; 0, 171, 422,
1,069 mg/kg-day (females)b
Diet
14 weeks
Nagao et al. (2000)
Rat (Sprague-Dawley);
20-25 parental rats/sex/
generation/group
(15-24 litters/generation/group)
0, 20, 100, 500 mg/kg-day
Gavage
Multigenerational study
FO males and females: exposure for
12 weeks prior to mating, 2 weeks
cohabitation, and until necropsy at
23 weeks of age (males) or
postpartum day (females); Fl
animals: exposure from weaning
until necropsy at PND 22
Note: Litters were culled to
8 rats/litter (4/sex, if possible) at
PND 4. At PND 22 (Fl) or PND 21
(F2), 2 rats/sex/litter were
sacrificed
Body weight (percent change compared to control)
mg/kg-day (M)
day 14
day 39
day 98
mg/kg-day (F)
day 14
day 39
day 98
0
0
0
0
0
0
0
0
151
-5
-4
-8*
171
2
2
0
381
-2
-8*
-8*
422
-1
2
-3
960
-5
-11*
-7*
1,069
-8*
-3
-5*
Percent change compared to control
mg/kg-day (M)
FO, parental
PND 0, Fl
PND 4, Fl
PND 7, Fl
PND 14, Fl
PND 21, Fl
PND 22, Fl (weanling
terminal body weight)
Body weight gain,
PNDs 22-91, Fl
Fl, adult (terminal
body weight)
PND 0, F2
PND 4, F2
PND 7, F2
PND 14, F2
PND 21, F2
mg/kg-day (F)
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
20
-2
0
3
2
0
1
-2
-3
-4
0
3
4
5
4
20
100
0
-6*
-5
-3
-1
-1
1
-7
-7*
0
5
5
4
3
100
500
-7*
-7*
-6
-6
-8*
-7*
-6*
-9
-13*
-3
0
-2
-5
-8
500
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
NTP (1997b)
Rat (F344); 15 males/group
0, 300, 900, 2,800, 8,300,
25,000 ppm
0, 30, 60, 180, 550, "high"
mg/kg-dayb
Diet
26 weeks
NTP (1997b)
Rat (F344); 15 males/group
0, 300, 2,800, 25,000 ppm
0, 20, 200, 2,200 mg/kg-dayb
Diet
10 weeks
NTP (1997b)
Rat (F344); 60 sex/group; interim
sacrifice of 10 rats/sex/group at
15 months
FO, parental
PND 0, Fl
PND 4, Fl
PND 7, Fl
PND 14, Fl
PND 21, Fl
PND 22, Fl (terminal
weanling body weight)
Body weight gain,
PNDs 22-91, Fl
Fl, adult (terminal
body weight)
PND 0, F2
PND 4, F2
PND 7, F2
PND 14, F2
PND 21, F2
Percent change compare<
mg/kg-day
terminal body weight
body weight change
Percent change compare<
mg/kg-day
terminal body weight
body weight change
Body weight (percent chc
mg/kg-day (M)
at interim sacrifice
at 69 weeks
Results
0
0
0
0
0
0
0
0
0
0
0
0
0
0
i to control
0 30
0 7
0 11
i to control
0
0
0
wge compared to
0
0
0
-2
2
3
1
1
1
1
0
1
2
4
4
9
4
60
10
16
20
0
-1
control)
L20
0
0
-1
-6*
-6
-5
-3
-2
-1
3
1
0
7
4
4
2
180
2
4
200
-1
-3
240
-6*
-1
1
-6*
-6
-6
-8*
-7*
-9*
2
0
-3
0
-4
-8
-12
550 ND
3 -30*
5 -44*
2,200
-29*
-45*
500
-9*
-6
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
0, 3,000, 6,000, 12,000 ppm
(males); 0, 6,000, 12,000,
24,000 ppm (females)
0, 120, 240, 500 mg/kg-day
(males)"; 0 300, 600,
1,200 mg/kg-day (females)b
Diet
2 years
Tvl et al. (2004)
Rat (CD); 30 FO and Fl parental
rats/sex/group
0, 750, 3,750, 11,250 ppm
0, 50, 250, 750 mg/kg-dayb
Diet
Multigenerational study
Exposure 10 weeks prior to mating
and through mating, gestation, and
lactation (females); or for 21 days
after mating (males)
Hazleton Laboratories (1958)
Rat (Sprague-Dawley);
10/s ex/group
0, 0.5, 2.0% (0, 5,000, 20,000 ppm)
0, 431, 1,551 mg/kg-day (males)";
0, 490, 1,765 mg/kg-day (females)"
Diet
90 days
Results
at study termination 0
mg/kg-day (F) 0
at interim sacrifice 0
at 69 weeks 0
at study termination 0
-2 -4
300 600
0 1
-1 -5
2 -3
-6
1,200
-23*
-25
-27
Body weight (percent change compared to control)
mg/kg-day (M) 0
adult FO at necropsy 0
adult Fl at necropsy 0
Fl offspring at 0
necropsy
F2 offspring at 0
necropsy
mg/kg-day (F) 0
50 250
-3 4
1 4
0 1
0 2
50 250
adult FO at necropsy 000
adult, Fl at necropsy 013
Fl offspring at 020
necropsy
F2 offspring at 033
necropsy
Note: No biologically significant changes were reported by the
authors for FO parental males or females.
750
-2
_9*
-18*
-11*
750
-4
-6*
-22*
-11*
study
Body weight (percent change compared to control)
mg/kg-day (M)
males
mg/kg-day (F)
females
0 431
0 -1
0 490
0 -1
1,551
-21*
1,765
-11
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Asoetal. (2005)
Rat(Crj:CD(SD)IGS);
24 rats/sex/generation/group
0, 100, 200, 400 mg/kg-day
Gavage
Multigenerational study
FO and Fl exposed for 4 weeks
prior to mating, through mating for
10 weeks, and until weaning of
offspring (females) or necropsy
after mating (males)
Betz et al. (2013)
Rat (Sprague-Dawley); male (NR)
0, 5, 10 ppm (equivalent to 0, 2,
4 mg/kg-day as indicated by study
authors)
Drinking water
Daily for 15 weeks (began PND 40
and ended PND 140)
Ahmad et al. (2014)
Rat (Albino);
PO, female (6/group)
0, 4, 20, 100 mg/kg
Gavage
GD 14 to parturition
Results
Body weight (percent change compared to control)
mg/kg-day (M)
FO, parental
Fl, parental
Fl, offspring at PND 21
F2, offspring at PND 21
mg/kg-day (F)
0 100
0 2
0 0
0 3
0 -4
0 100
200
4
-2
0
0
200
400
-1
-4
-2
-7
400
FO, parental 0061
Fl, parental 0461
Note: The study authors indicated that body weights were significantly
lowered at PND 0 at 100 mg/kg-day and higher in Fl male offspring and at
100 and 400 mg/kg-day in F2 males and females. No significant effects
were observed in Fl female offspring (data presented graphically).
No significant change in
body weight reported by the
study authors
Body weight (percent change compared to control)
mg/kg-day
Fl adult male
0 4
0 -1
20
-2*
100
_4*
Urinary bladder histopathology
NTP (1997b)
Rat (F344); 60 sex/group;
10 rats/sex/group sacrificed at
15 mo
0, 3,000, 6,000, 12,000 ppm
(males); 0, 6,000, 12,000,
24,000 ppm (females)
Percent incidence
mg/kg-day (M)
transitional epithelium;
hyperplasia
hemorrhage
inflammation,
suppurative
0 120
0 0
0 2
0 4
240
0
0
0
500
4
0
0
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
0, 120, 240, 500 mg/kg-day
(males)"; 0 300, 600,
1,200 mg/kg-day (females)b
Diet
2 years
BIBRA (1978)
Rat (Wistar); 27/sex/group or
45/sex/group (control); interim
sacrifices of 9 controls/sex/group
and 6 treated rats/sex/group at
2 and 6 weeks
0, 2,000, 5,000, 12,000 ppm
0, 151, 381, 960 mg/kg-day
(males)"
0, 171, 422, 1,069 mg/kg-day
(females)"
Diet
14 weeks
NTP (1997b)
Rat (F344); 15 males/group
0, 300, 2,800, 25,000 ppm
0, 20, 200, 2,200 mg/kg-dayb
Diet
10 weeks
NTP (1997b)
Rat (F344); 15 males/group
0, 300, 900, 2,800, 8,300,
25,000 ppm
0, 30, 60, 180, 550, "high"
mg/kg-dayb
Results
transitional epithelium; None reported (incidence data not provided)
papilloma
adenocarcinoma, 0020
metastatic, intestine
large, colon
mg/kg-day (F) 0 300
transitional epithelium; 8 0
hyperplasia
transitional epithelium; 2 0
papilloma
edema 2 0
hemorrhage 0 0
600 1,200
2 20*
0 4
2 0
2 0
Percent incidence
mg/kg-day (M) 0 151
proteinaceous deposits; 11 NE
6 weeks
proteinaceous deposits; 4 NE
14 weeks
hyperplasia; 14 weeks 0 NE
mg/kg-day (F) 0 171
proteinaceous deposits; 0 NE
6 weeks
proteinaceous deposits; 0 NE
14 weeks
hyperplasia; 14 weeks 0 NE
No significant treatment-related effects reported
animals (quantitative data not reported).
No significant treatment- related effects reported
animals (quantitative data not reported).
381 960
NE 0
NE 0
NE 7
422 1,069
NE 0
NE 0
NE 0
in control or high-dose
in control or high-dose
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Diet
26 weeks
NTP (1997a)
Rat (F344); 50-60/sex/group;
interim sacrifice of
10 rats/sex/group at 15 months
0, 12,000 ppm (males); 0,
24,000 ppm (females)
0, 500 mg/kg-day (males)b; 0,
1,200 mg/kg-day (females)b
Diet
4 exposure protocols: ad libitum
feeding, weight-matched controls,
restricted feed (2 years), and
restricted feed (lifetime) 2 years to
lifetime
Results
Ad libitum feeding, weight-matched protocol (percent incidence)
mg/kg-day (M) 0
(ad libitum)
transitional epithelium 0
hyperplasia
mg/kg-day (F) 0
(ad libitum)
transitional epithelium 8
hyperplasia
0 (weight-
matched)
0
0 (weight-
matched)
0
500
4
1,200
20
Feed-restricted, 2 years or lifetime (percent incidence)
mg/kg-day (M) 0 500 0 500
(2 years) (2 years) (lifetime) (lifetime)
transitional epithelium 2402
hyperplasia
mg/kg-day (F) 0 1,200 0 1,200
(2 years) (2 years) (lifetime) (lifetime)
transitional epithelium 0 28 0 32
hyperplasia
1
2
3
4
5
6
7
8
*Statistically significant (p <0.05) relative to controls based on statistics performed by the study authors.
aPercent change compared to control calculated as 100 x ((treated value - control value) 4- control value).
Calculated as follows: [% in diet x intake food/water (mg)] 4 body weight (kg) = mg/kg-day.
ND = not determined; NE = not examined
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
NTP, 1997b; lOwks;
male F344 rats
Hazlcton Laboratories, 1958; 90 d;
male S-D rats
Hazleton Laboratories, 1958; 90 d;
female S-D rats
Bl BRA 1978; 14wks;
male Wistar rats
Bl BRA, 1978; 14wks;
female Wistar rats
NTP,1997b;26wks;
male F344 rats
Betz ct al., 2013; PND40-PND140;
male S-D rats
Ahmad, 2014; GD 14-parturition;
Fl adult male albino rats
Asocial., 2005; 2-gcn;
S-D rats
Nagao et al., 2000; 2-gen;
Fl (adult terminal) male S-D rats
Nagao ct al., 2000; 2-gen;
Fl (adult terminal) female S-D rats
Ty I eta I., 2 004; 2-gen;
FO at necropsy male and female CD rats
Tyl ct al., 2004; 2-gcn; males females Fl adult, Fl offspring,
F2 offspring at necropsy CD rats
NTP, 1997b; interim sac
male F344 rats
NTP, 1997b; interim sac
female F344rats
NTP, 1997b; 2yr study termination
male F344 rats
NTP, 1997b; 2yr study termination
female F344rats
NTP,1997b;10wks;
male F344 rats
Bl BRA 1978; 14wks;
male Wistar rats
Bl BRA 1978; 14wks;
female Wistar rats
NTP,1997b;26wks;
male F344 rats
NTP, 1997b; 2 yr; transitional epithelial hypcrplasia;
fcmaleF344rats
NTP, 1997b; 2 yr; transitional epithelial hyplerplasia
male F344 rats
NTP 1997a; 2 yr or lifetime; ad-lib wt matched, feed;
male F344 rats
NTP 1997a; 2 yr or lifetime; ad-lib wt matched, feed;
femalcF344rats
Q B—
Q—B B B
Q—B
B-
o—B—a
B
-B B
Q—B—B
B—B—Q
D
D
B—B—B
B
= statistically significant
- not statistically significant
10 100 1000
Dose (mg/kg-day)
10000
2
3
Figure 3-21. Exposure response array of other health effects following oral
exposure to BBP.
This document is a draft for review purposes only and does not constitute Agency policy.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 3.3.12. BBP Metabolite Studies
2 Table 3-34. Evidence pertaining to toxicity effects in animals following
3 exposure to BBP metabolites
Reference and study design
Results by endpoint"
Developmental body weight
Ema et al. (1996a)
MBzP
Rat (Wistar); PO, female (11-15/group)
0, 375, 500, 625 mg/kg-day
Gavage
GDs 7-9; dams sacrificed on GD 20
Ema et al. (1996a)
MBzP
Rat (Wistar); PO, female (10-12/group)
0, 250, 375, 500, 625 mg/kg-day
Gavage
GDs 10-12; dams sacrificed on GD 20
Ema et al. (1996a)
MBzP
Rat (Wistar); PO, female (10-17/group)
0, 250, 375, 500, 625 mg/kg-day
Gavage
GDs 13-15; dams sacrificed on GD 20
Ema et al. (1996b)
MBzP
Rat (Wistar); PO, female (10-14/group)
0, 250, 313, 375, 438, 500 mg/kg-day
Gavage
GDs 7-15; dams sacrificed on GD 20
Body weight of live fetuses (g, litter mean ± SD)
Dose 0 375 500 625
female 3.93 3.59 3.43 2.97 (±0.27)*
(±0.13) (±0.18)* (±0.15)*
male 4.2 (±0.18) 3.91 (±0.17) 3.67 3.38 (±0.18)*
(±0.28)*
Body weight of live fetuses (g, litter mean ± SD)
Dose 0 250 375 500 625
female 3.77 3.73 3.78 3.45 2.8 (±0.21)*
(±0.1) (±0.22) (±0.21) (±0.43)*
male 4.07 4.04 4.06 3.74 3.42
(±0.13) (±0.23) (±0.23) (±0.3) (±0.76)*
Body weight of live fetuses (g, litter mean ± SD)
Dose 0 250 375 500 625
female 3.86 3.8 3.77 3.81 3.59 (±0.22)
(±0.12) (±0.26) (±0.13) (±0.19)
male 4.1 4.03 3.97 4 (±0.13) 4.17
(±0.12) (±0.22) (±0.22)
Body weight of live fetuses (g, litter mean ± SD)
Dose 0 250 313 375 438 500
female 3.84 3.64 3.65 3.52 3.57 3.35
(±0.12) (±0.28) (±0.19) (±0.24)* (±0.21)* (±0.2)*
male 4.08 3.97 3.93 3.84 3.78 3.59
(±0.21) (±0.26) (±0.25) (±0.15) (±0.3)* (±0.12)*
This document is a draft for review purposes only and does not constitute Agency policy,
3-196 DRAFT—DO NOT CITE OR QUOTE
-------
Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results by endpoint"
Ema et al. (1996c)
MBP
Rat (Wistar); PO, female (10-11/group)
0, 500, 625, 750 mg/kg-day
Gavage
GDs 7-9; dams sacrificed on GD 20
Body weight of live fetuses (g, litter mean ± SD)
Dose
0
500
625
750
female
male
3.77 (±0.16)
4.05 (±0.16)
3.46
(±0.09)*
3.74
(±0.13)*
3.26 3.15 (±0.26)*
(±0.17)*
3.58 3.52 (±0.17)*
(±0.17)*
Ema et al. (1996c)
MBP
Rat (Wistar); PO, female (10-14/group)
0, 500, 625, 750 mg/kg-day
Gavage
GDs 10-12; dams sacrificed on GD 20
Body weight of live fetuses (g, litter mean ± SD)
Dose
0
500
625
750
female 3.77 (±0.16) 3.53 (±0.35) 3.53 (±0.26) 2.95 (±0.53)*
male 4.05 (±0.16) 3.78 (±0.3)* 3.81 (±0.19) 3.1 (±0.4)*
Ema et al. (1996c)
MBP
Rat (Wistar); PO, female (10-15/group)
0, 500, 625, 750 mg/kg-day
Gavage
GDs 13-15; dams sacrificed on GD 20
Body weight of live fetuses (g, litter mean ± SD)
Dose
0
500
625
750
female 3.77 (±0.16) 3.77 (±0.17) 3.68 (±0.17) 3.5 (±0.12)
male 4.05 (±0.16) 3.97 (±0.18) 3.9 (±0.26) 3.81 (± 0.04)
Ema and Mivawaki (2001)
MBP
Rat (Wistar); PO, female (16/group)
0, 250, 500, 750,1,000 mg/kg-day
Gavage
GDs 0-8, with outcomes determined on
GD20
Body weight of live fetuses (g, litter mean ± SD)
Dose
0
250
500
750
1,000
female
male
3.17
(±0.22)
3.35
(±0.25)
3.15
(±0.15)
3.42
(±0.1)
2.8
(±0.3)*
3.01
(±0.36)*
2.58
(±0.23)*
2.71
(±0.3)*
2.32
(±0.29)*
2.47
(±0.29)*
Ema et al. (2003)
MBzP
Rat (Wistar); PO, female (16/group)
0,167, 250, 375 mg/kg-day
Gavage
GDs 15-17; dams sacrificed on GD 21
Body weight of live fetuses (g, litter mean ± SD)
Dose
0
167
250
375
female 4.63 (±0.2) 4.58 (±0.2) 4.39 (±0.24) 3.67 (±0.56)*
male 4.95 (±0.25) 4.95 (±0.24) 4.7 (±0.3) 3.82 (±0.65)*
This document is a draft for review purposes only and does not constitute Agency policy,
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-------
Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Saillenfait et al. (2003)
MBP
Rat (Sprague-Dawley); PO, female
(14-15/group)
0, 1.8, 3.6, 5.4 mmol/kg (equivalent to
0, 560, 1,120, 1,690 mg/kg as calculated
by study authors)
Gavage
GD 10; dams sacrificed on GD 21
Saillenfait et al. (2003)
MBzP
Rat (Sprague-Dawley); PO, female
(12-14/group)
0, 0.9, 1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 280, 560, 1,120, 1,690 mg/kg as
calculated by study authors)
Gavage
GD 10; dams sacrificed on GD 21
Saillenfait et al. (2003)
MBP
Mouse (OF-1); PO, female (24-25/group)
0, 0.9, 1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 280, 560, 1,120, and 1,690 mg/kg
as calculated by study authors)
Gavage
GD 8; dams sacrificed on GD 18
Saillenfait etal. (2003)
MBzP
Mouse (OF-1); PO, female (20-23/group)
0, 0.9, 1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 280, 560, 1,120, 1,690 mg/kg as
calculated by study authors)
Gavage
GD 8; dams sacrificed on GD 18
Body weight of
Dose
male and 5
female
Body weight of
Dose
male and !
female (±
Body weight of
Dose
male and
female (±
Body weight of
Dose
male and
female (±
Results by endpoint"
live fetuses (g, litter mean ± SE)
0 560 1,120
28 (±0.07) 5.15 (±0.16) 5.19 (±0.15)
live fetuses (g, litter mean ± SE)
0 280 560 1,120
5.04 5.25 5.14 4.82
0.18) (±0.2) (±0.18) (±0.1)
live fetuses (g, litter mean ± SE)
0 280 560 1,120
L.19 1.16 1.23 1.14
0.02) (±0.03) (±0.05) (±0.03)
live fetuses (g, litter mean ± SE)
0 280 560 1,120
L.21 1.24 1.21 1.13
0.03) (±0.05) (±0.02) (±0.02)
1,690
5.25 (±0.16)
1,690
4.93
(± 0.06)
1,690
1.04
(± 0.04)*
1,690
1.11
(± 0.07)
This document is a draft for review purposes only and does not constitute Agency policy,
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-------
Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results by endpoint"
Developmental embryotoxic effects
Ema et al. (1996a)
MBzP
Rat (Wistar); PO, female (11-15/group)
0, 375, 500, 625 mg/kg-day
Gavage
GDs 7-9; dams sacrificed on GD 20
Ema et al. (1996a)
MBzP
Rat (Wistar); PO, female (10-12/group)
0, 250, 375, 500, 625 mg/kg-day
Gavage
GDs 10-12; dams sacrificed on GD 20
Percent postimplantation loss per litter (mean)
Dose 0 375 500 625
10.2 18.9 25.7* 90.6*
Maternal adjusted weight gain (g, mean ± SD)
Dose 0 375 500 625
53 (±12) 37 (±11)* 42 (±9) 23 (± 13)*
Maternal food consumption
Significant decrease in all treatment groups during treatment, but only
remained significantly lower after treatment in the 625 mg/kg group
Number of litters totally resorbed
Dose 0 375 500 625
0009*
Number of live fetuses per litter (mean ± SD)
Dose 0 375 500 625
12.7 (±1) 11.6 (±2.2) 10.6 (±2.8) 1.4 (± 2.6)*
Number of resorptions and dead fetuses per litter (mean ± SD)
Dose 0 375 500 625
1.5 (±0.8) 2.7 (±2) 3.6 (±2.7) 12.8 (± 2.5)*
Sex ratio of live fetuses (male/female)
No significant change in sex ratio (male/female): 67/73 (control),
63/65 (375 mg/kg), 56/61 (500 mg/kg), 9/8 (625 mg/kg)
Percent postimplantation loss per litter (mean)
Dose 0 250 375 500 625
15.5 16.9 15.3 54.8* 90.4*
Maternal adjusted weight gain (g, mean ± SD)
Dose 0 250 375 500 625
47 (±10) 25 (±7)* 41 (±6) 29 (± 12)* 31 (± 9)*
Maternal food consumption
Significant decrease in food consumption during treatment as well as
the remainder of gestation for all dose groups
Number of litters totally resorbed
This document is a draft for review purposes only and does not constitute Agency policy,
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-------
Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema et al. (1996a)
MBzP
Rat (Wistar); PO, female (10-17/group)
0, 250, 375, 500, 625 mg/kg-day
Gavage
GDs 13-15; dams sacrificed on GD 20
Results by endpoint"
Dose 0 250 375
000
500 625
3 7*0
Number of live fetuses per litter (mean ± SD)
Dose 0 250 375
11.8 11.5 11.5
(±1.8) (±2.1) (±1.6)
500 625
6.6 1.4 (±2.5)*
(±5.8)*
Number of resorptions and dead fetuses per litter (mean ± SD)
Dose 0 250 375
2 (±1.9) 2.4 (±1.6) 2.2 (±1.9)
500 625
8.2 13.5
(±5.7)* (±2.4)*
Sex ratio of live fetuses (male/female)
No significant change in sex ratio (male/female): 62/68 (control),
51/64 (250 mg/kg), 54/61 (375 mg/kg), 44/29 (500 mg/kg),
7/7 (625 mg/kg)
Percent postimplantation loss per litter (mean)
Dose 0 250 375
18.8 12.8 28.3
500 625
61.4* 94.2*
Maternal adjusted weight gain (g, mean ± SD)
Dose 0 250 375
47 (± 7) 29 (± 16)* 23 (± 7)*
500 625
29 (± 14)* 25 (± 11)*
Maternal food consumption
Significant decrease in food consumption in all dose groups during
treatment, but only continued to be low throughout gestation in the
625 mg/kg group
Number of litters totally resorbed
Dose 0 250 375
000
500 625
5* 11*
Number of live fetuses per litter (mean ±SD)
Dose 0 250 375
11.4 13.3 10 (±3.7)
(±2.2) (±1.2)
500 625
5.6 0.8 (±2.5)*
(±5.6)*
Number of resorptions and dead fetuses per litter (mean ±SD)
Dose 0 250 375
2.5 (±1.5) 2 (±1.2) 3.8 (±3.2)
500 625
8.9 13.5
(±5.6)* (±3.2)*
Sex ratio of live fetuses (male/female)
This document is a draft for review purposes only and does not constitute Agency policy,
3-200 DRAFT—DO NOT CITE OR QUOTE
-------
Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema et al. (1996b)
MBzP
Rat (Wistar); PO, female (10-14/group)
0, 250, 313, 375, 438, 500 mg/kg-day
Gavage
GDs 7-15; dams sacrificed on GD 20
Ema et al. (1996c)
MBP
Rat (Wistar); PO, female (10-11/group)
0, 500, 625, 750 mg/kg-day
Gavage
GDs 7-9; dams sacrificed on GD 20
Results by endpoint"
No significant change in sex ratio (male/female): 64/61 (control),
71/75 (250 mg/kg), 60/40 (375 mg/kg), 38/41 (500 mg/kg),
5/5 (625 mg/kg)
Percent postimplantation loss per litter (mean)
Dose 0 250 313 375 438 500
15.8 8.3 18.7 23.8 36.6* 82.3*
Food consumption during pregnancy
Significantly lower that control on GDs 7-15 at >250 mg/kg and on
GDs 15-20 at 500 mg/kg
Live fetuses per litter (mean ±SD)
Dose 0 250 313 375 438 500
11.8 12.9 11.6 10.8 9.2 2.4 (± 3.3)*
(±2.1) (±2.3) (±1.5) (±2.9) (±5.2)
Number of resorptions and dead fetuses per litter (mean ±SD)
Dose 0 250 313 375 438 500
2.2 1.2 2.8 3.4 5 (± 4.4) 12.2
(±1.5) (±1.5) (±1.7) (±3) (±4.2)*
Number of litters totally resorbed
Dose 0 250 313 375 438 500
00002 6*
Sex ratio of live fetuses
Sex ratios (male/female): 54/76 (control), 65/64 (250 mg/kg),
74/65 (313 mg/kg), 55/53 (375 mg/kg), 55/65 (438 mg/kg),
13/11 (500 mg/kg)
Weight gain during pregnancy
Significantly lower than control on GDs 7-15 at >313 mg/kg, and on
GDs 15-20 at 500 mg/kg; adjusted weight gain significantly lower that
control at 500 mg/kg
Adjusted maternal body weight gain
No significant change
Maternal food intake during pregnancy (g, mean ± SD)
Dose 0 500 625 750
384 (±22) 366 (±27) 355 (± 20)* 336 (± 30)*
Number of litters totally resorbed
Dose 0 500 625 750
001 3
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results by endpoint"
Number of live fetuses per litter (mean ± SD)
Dose
0
500
625
750
12.3 (±2.4) 12.1 (±1.9) 10.3 (±4.1) 5.9 (±4.5)*
Percent postimplantation loss per litter (mean)
Dose
0
500
625
750
13.3
18.4
27.8*
57.7*
Sex ratio of live fetuses
No significant change in sex ratio of live fetuses (male/female):
59/64 (control), 53/68 (500 mg/kg), 46/66 (625 mg/kg),
30/35 (750 mg/kg)
Ema et al. (1996c)
MBP
Rat (Wistar); PO, female (10-14/group)
0, 500, 625, 750 mg/kg-day
Gavage
GDs 10-12; dams sacrificed on GD 20
Adjusted maternal body weight gain
No significant change
Maternal food intake during pregnancy (g, mean ± SD)
Dose
0
500
625
750
384 (±22) 387 (±16) 370 (± 27) 349 (± 28)*
Number of litters totally resorbed
Dose
0
500
625
750
0
0
Number of live fetuses per litter (mean ±SD)
Dose
0 500 625 750
12.3 (±2.4) 11.2 (±2.8) 7.5 (± 3.8)* 1.8 (± 3.3)*
Percent postimplantation loss per litter (mean)
Dose
0
500
625
750
13.3
24.6
46.4*
86.9*
Sex ratio of live fetuses
No significant change in sex ratio of live fetuses (male/female):
59/64 (control), 58/54 (500 mg/kg), 40/42 (625 mg/kg),
15/10 (750 mg/kg)
Ema et al. (1996c)
MBP
Rat (Wistar); PO, female (10-15/group)
0, 500, 625, 750 mg/kg-day
Gavage
GDs 13-15; dams sacrificed on GD 20
Adjusted maternal body weight gain
No significant change
Maternal food intake during pregnancy (g, mean ± SD)
Dose
0
500
625
750
384 (±22) 372 (±22) 370 (± 18) 350 (± 21)*
Number of litters totally resorbed
Dose
0
500
625
750
This document is a draft for review purposes only and does not constitute Agency policy,
3-202 DRAFT—DO NOT CITE OR QUOTE
-------
Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema and Mivawaki (2001)
MBP
Rat (Wistar); PO, female (16/group)
0, 250, 500, 750, 1,000 mg/kg-day
Gavage
GDs 0-8 with outcomes determined on
GD20
Results by endpoint"
PO, female 0 0 2 12*
Number of live fetuses per litter (mean ±SD)
Dose 0 500 625 750
12.3 (±2.4) 8.6 (±3.5) 4.6 (± 3.4)* 0.6 (± 1.5)*
Percent postimplantation loss per litter (mean)
Dose 0 500 625 750
13.3 34.7* 66.8* 95.5*
Sex ratio of live fetuses
No significant change in sex ratio of live fetuses (male/female):
59/64 (control), 55/40 (500 mg/kg), 25/26 (625 mg/kg),
3/6 (750 mg/kg)
Adjusted maternal weight gain (g, mean ± SD)
Dose 0 250 500 750 1,000
33 (± 13) 38 (± 9) 31 (± 10) 37 25 (± 12)
(±13)
Maternal adjusted weight gain (body weight gain excluding uterus) was
not statistically significantly different among treated and controls
Number of live fetuses per litter (mean ±SD)
Dose 0 250 500 750 1,000
14.1 13.7 13.9 (±2.4) 12.7 10.8
(±1.6) (±2.7) (±2.7) (±3.7)*
Number of resorptions and dead fetuses per litter (mean ±SD)
Dose 0 250 500 750 1,000
1.4 (±1.5) 1(±1) 1.7 (±1.7) 2.4 3.7 (±3.1)*
(±2)
Percent postimplantation loss per litter (mean)
Dose 0 250 500 750 1,000
9.1 6.4 11.3 15.9 26.3*
Postimplantation loss = (number of resorptions and dead fetuses/
number of implantations) x 100
Percent preimplantation loss per female (mean)
Dose 0 250 500 750 1,000
5.9 8.7 9.8 19.2 20.2*
n = number of pregnant females; preimplantation loss = ((number of
corpora lutea - number of implantations)/number of corpora lutea) x
100
This document is a draft for review purposes only and does not constitute Agency policy,
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-------
Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results by endpoint"
Percent preimplantation loss per litter
Dose
0
250
500
750
1,000
5.9
8.7
3.7
7.6
8.7
n = number of litters; preimplantation loss = ((number of corpora lutea
- number of implantations)/number of corpora lutea) x 100
Sex ratio of live fetuses (male/female)
No significant difference in sex ratio (males/females):
121/104 (controls), 120/99 (250 mg/kg), 108/100 (500 mg/kg),
98/80 (750 mg/kg), 77/74 (1,000 mg/kg)
Ema et al. (2003)
MBzP
Rat (Wistar); PO, female (16/group)
0,167, 250, 375 mg/kg-day
Gavage
GDs 15-17; dams sacrificed on GD 21
Body weight gain during pregnancy
Maternal body weight gain significantly decreased on GDs 15-18 at
>167 mg/kg and GDs 18-21 at >250 mg/kg; adjusted weight gain
significantly reduced at >250 mg/kg
Food consumption during pregnancy
Maternal food consumption significantly decreased on GDs 15-18 at
>167 mg/kg and GDs 18-21 at >250 mg/kg
Number of litters totally resorbed
Dose
0
167
250
375
0
0
Number of live fetuses per litter (mean ±SD)
Dose
0
167
250
375
14.1 (±1.8) 12.8 (±1.9) 13.8 (±0.8) 13.2 (± 1.9)
Number of resorptions and dead fetuses per litter (mean ±SD)
Dose
0
167
250
375
1.4 (±1.1) 0.7 (±0.9) 1.1 (±0.8) 1.3 (±1.9)
Percent postimplantation loss per litter (mean)
Dose
0
167
250
375
9.7
5.3
8.1
10.9
Sex ratio of live fetuses (male/female)
No significant change in sex ratio of live fetuses (male/female):
105/101 (control), 109/96 (167 mg/kg), 107/114 (250 mg/kg),
117/94 (375 mg/kg)
Saillenfait et al. (2003)
MBP
Rat (Sprague-Dawley); PO, female
(14-15/group)
Number of live fetuses per litter (mean ±SD)
Dose
0
560
1,120
1,690
13.46 (±0.77) 13.92 (±0.55) 13.5 (±0.69) 12.77 (± 0.67)
Percent postimplantation loss per litter (mean ± SE)
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
0, 1.8, 3.6, 5.4 mmol/kg (equivalent to
0, 560, 1,120, 1,690 mg/kg as calculated
by study authors)
Gavage
GD 10; sacrificed on GD 21
Saillenfait et al. (2003)
MBzP
Rat (Sprague-Dawley); PO, female
(12-14/group)
0, 0.9, 1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 280, 560, 1,120, 1,690 mg/kg as
calculated by study authors)
Gavage
GD 10; sacrificed on GD 21
Saillenfait et al. (2003)
MBP
Mouse (OF-1); PO, female (24-25/group)
0, 0.9, 1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 280, 560, 1,120, 1,690 mg/kg as
calculated by study authors)
Gavage
GD 8; sacrificed on GD 18
Saillenfait et al. (2003)
MBzP
Mouse (OF-1); PO, female (20-23/group)
0, 0.9, 1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 280, 560, 1,120, 1,690 mg/kg as
calculated by study authors)
Results by endpoint"
Dose 0 560 1,120
2.1 (±1.08) 4.38 (±1.77) 1.79 (± 1.28)
Percent resorptions per litter (mean ± SE)
Dose 0 560 1,120
2.1 (±1.08) 4.38 (±1.77) 1.79 (± 1.28)
Number of live fetuses per litter (mean ±SE)
Dose 0 280 560 1,120
13.77 12.83 13.67 14.17
(±1.08) (±1.15) (±1.14) (±0.55)
Percent postimplantation loss per litter (mean ± SE)
Dose 0 280 560 1,120
1.61 4.1 7.44 6.2
(±0.87) (±1.67) (±4.07) (±1.81)
Percent resorptions per litter (mean ± SE)
Dose 0 280 560 1,120
1.61 4.1 7.44 6.2
(±0.87) (±1.67) (±4.07) (±1.81)
Number of live fetuses per litter (mean ±SE)
Dose 0 280 560 1,120
12.35 12.38 6.64 2.32
(±0.88) (±0.71) (±0.91)* (±0.69)*
Percent postimplantation loss per litter (mean ± SE)
Dose 0 280 560 1,120
9.59 11.25 40.83 83.31
(±2.76) (±2.5) (±6.22)* (±5.03)*
Percent resorptions per litter (mean ± SE)
Dose 0 280 560 1,120
9.3 10.21 40.15 82.21
(±2.76) (±2.48) (±6.17)* (±4.96)*
Number of live fetuses per litter (mean ±SE)
Dose 0 280 560 1,120
11.07 11.25 12.11 12.82
(±1.4) (±1.1) (±0.88) (±0.54)
Percent postimplantation loss per litter (mean ± SE)
1,690
6.1 (±1.99)
1,690
6.1 (±1.99)
1,690
13.75 (± 1.7)
1,690
8.93 (±8.93)
1,690
8.93 (±8.93)
1,690
2.33
(±0.58)*
1,690
82.42
(±4.31)*
1,690
80.66
(±4.45)*
1,690
7.5 (± 1.58)
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Gavage
GD 8; sacrificed on GD 18
Results by endpoint"
Dose 0 280 560 1,120
14.17 14.69 7.8 (±2.51) 12.24
(±7.15) (±3.16) (±2.57)
1,690
47.38
(± 10.45)*
Resorptions per litter (mean ± SE)
Dose 0 280 560 1,120
14.17 14.69 7.8 (±2.51) 10.99
(±7.15) (±3.16) (±2.39)
1,690
45.95
(± 10.35)*
Developmental teratological effects
Ema et al. (1996a)
MBzP
Rat (Wistar); PO, female (11-15/group)
0, 375, 500, 625 mg/kg-day
Gavage
GDs 7-9; dams sacrificed on GD 20
Ema et al. (1996a)
MBzP
Rat (Wistar); PO, female (10-12/group)
0, 250, 375, 500, 625 mg/kg-day
Gavage
GDs 10-12; dams sacrificed on GD 20
Number of fetuses with external malformations
Dose 0 375 500
001
625
0
Number of fetuses with internal malformations
Dose 0 375 500
026
Dilation of renal pelvis
625
10
Number of fetuses with skeletal malformations
Dose 0 375 500
625
267 10
Mainly fusion or absence of ribs, fusion or absent cervical/thoracic/
lumbar vertebral arches or bodies
Number of fetuses with external malformations
Dose 0 250 375 500
0000
625
0
Number of fetuses with internal malformations
Dose 0 250 375 500
1000
625
0
Number of fetuses with skeletal malformations
Dose 0 250 375 500
110 1
625
0
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema et al. (1996a)
MBzP
Rat (Wistar); PO, female (10-17/group)
0, 250, 375, 500, 625 mg/kg
Gavage
GDs 13-15; dams sacrificed on GD 20
Ema et al. (1996b)
MBzP
Rat (Wistar); PO, female (10-14/group)
0, 250, 313, 375, 438, 500 mg/kg-day
Gavage
GDs 7-15; dams sacrificed on GD 20
Ema et al. (1996c)
MBP
Rat (Wistar); PO, female (10-11/group)
0, 500, 625, 750 mg/kg-day
Gavage
GDs 7-9; dams sacrificed on GD 20
Results by endpoint"
Number of fetuses with external malformations
Dose 0 250 375 500 625
1 0 3 11 1
Mainly cleft palate
Number of fetuses with internal malformations
Dose 0 250 375 500 625
0000 0
Number of fetuses with skeletal malformations
Dose 0 250 375 500 625
0 3 6 13 3
Mainly fusion of the sternebrae
Number of fetuses with external malformations
Dose 0 250 313 375 438 500
01 1 0 13 1
Mainly cleft palate
Number of fetuses with internal malformations
Dose 0 250 313 375 438 500
00 7 6 10 5
Included mainly dilatation of renal pelvis and hypoplasia of the kidney
Number of fetuses with skeletal malformations
Dose 0 250 313 375 438 500
0 1 8 12 13 6
Included mainly fusion or absence of cervical vertebral arches, fusion or
absence of ribs or sternebrae
Number of fetuses with external malformations
Dose 0 500 625 750
005 4
Mainly cleft palate and agenesis of the lower body
Number of fetuses with internal malformations
Dose 0 500 625 750
003 0
Dilation of renal pelvis and hypoplasia of kidney
Number of fetuses with skeletal malformations
Dose 0 500 625 750
This document is a draft for review purposes only and does not constitute Agency policy,
3-207 DRAFT—DO NOT CITE OR QUOTE
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema et al. (1996c)
MBP
Rat (Wistar); PO, female (10-14/group)
0, 500, 625, 750 mg/kg-day
Gavage
GDs 10-12; dams sacrificed on GD 20
Ema et al. (1996c)
MBP
Rat (Wistar); PO, female (10-15/group)
0, 500, 625, 750 mg/kg-day
Gavage
GDs 13-15; dams sacrificed on GD 20
Saillenfait et al. (2003)
MBP
Rat (Sprague-Dawley); PO, female
(14-15/group)
0, 1.8, 3.6, 5.4 mmol/kg (equivalent to
0, 560, 1,120, 1,690 mg/kg as calculated
by study authors)
Gavage
GD 10; sacrificed on GD 21
Results by endpoint"
1 10 10
Mainly fusion and/or absence of cervical vertebral arches
14
Number of fetuses with external malformations
Dose 0 500 625
000
750
1
Number of fetuses with internal malformations
Dose 0 500 625
031
Dilation of the renal pelvis
750
0
Number of fetuses with skeletal malformations
Dose 0 500 625
100
750
0
Number of fetuses with external malformations
Dose 0 500 625
0 1 16
Mainly cleft palate
750
9
Number of fetuses with internal malformations
Dose 0 500 625
000
750
0
Number of fetuses with skeletal malformations
Dose 0 500 625
1 6 10
Mainly fusion of the sternebrae
750
5
Percent of malformed fetuses (%)
Dose 0 560 1,120
000
Statistical significance not evaluated
1,690
0
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results by endpoint"
Saillenfait et al. (2003)
MBzP
Rat (Sprague-Dawley); PO, female
(12-14/group)
0, 0.9,1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 280, 560,1,120,1,690 mg/kg as
calculated by study authors)
Gavage
GD 10; sacrificed on GD 21
Percent of malformed fetuses
Dose
280
560
1,120
Oa 0.6
Statistical significance not evaluated
0
1,690
0
Saillenfait et al. (2003)
MBP
Mouse (OF-1); PO, female (24-25/group)
0, 0.9,1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 280, 560,1,120, and 1,690 mg/kg
as calculated by study authors)
Gavage
GD 8; sacrificed on GD 18
Percent of malformed fetuses
Dose
0
280
560
1,120
0 0.4 2
Statistical significance not evaluated
9.8
1,690
34.7
Saillenfait et al. (2003)
MBzP
Mouse (OF-1); PO, female (20-23/group)
0, 0.9,1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 280, 560,1,120, and 1,690 mg/kg
as calculated by study authors)
Gavage
GD 8; sacrificed on GD 18
Percent of malformed fetuses
Dose
0
280
560
1,120
000
Statistical significance not evaluated
3.2
1,690
22.9
Female reproductive effects
Ema et al. U996a)
MBzP
Rat (Wistar); PO, female (11-15/group)
0, 375, 500, 625 mg/kg-day
Gavage
GDs 7-9; dams sacrificed on GD 20
Number of implantations per litter (mean ± SD)
Dose
0
375
500
625
14.2 (±1) 14.4 (±1.5) 14.3 (±1.1) 14.2 (± 1.6)
Ema et al. U996a)
MBzP
Rat (Wistar); PO, female (10-12/group)
0, 250, 375, 500, 625 mg/kg
Number of implantations per litter (mean ± SD)
Dose
0
250
375
500
625
14 (±0.6) 13.9 13.7 14.8 (±1.7)
(± 1.4) (± 1.8)
14.9 (± 1.4)
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results by endpoint"
Gavage
Dams dosed on GD 10-12 and sacrificed
on GD20
Ema et al. (1996a)
MBzP
Rat (Wistar); PO, female (10-17/group)
0, 250, 375, 500, 625 mg/kg-day
Gavage
GDs 13-15; dams sacrificed on GD 20
Number of implantations per litter (mean ± SD)
Dose
0
250
375
500
625
13.9 (±1) 15.2 13.8 (±1) 14.6 (±0.8) 14.2 (± 1.5)
(± 1-5)
Ema et al. (1996b)
MBzP
Rat (Wistar); PO, female (10-14/group)
0, 250, 313, 375, 438, 500 mg/kg-day
Gavage
GDs 7-15; dams sacrificed on GD 20
Number of implantations per litter (mean ± SD)
Dose
0
250
313
375
438
500
14 14.1 14.3 14.2
(±1.3) (±2.1) (±1.7) (±0.9)
14.2 14.6 (± 1.1)
(±1.5)
Ema et al. (1996c)
MBuP
Rat (Wistar); PO, female (10-11/group)
0, 500, 625, 750 mg/kg-day
Gavage
GDs 7-9; dams sacrificed on GD 20
Number of implantations per litter (mean ± SD)
Dose
0
500
625
750
14.2 (±1.1) 15 (±1.3) 14.2 (±1.3) 14.5 (± 1.9)
Ema et al. (1996c)
MBP
Rat (Wistar); PO, female (10-14/group)
0, 500, 625, 750 mg/kg
Gavage
GDs 10-12; dams sacrificed on GD 20
Number of implantations per litter (mean ± SD)
Dose
0
500
625
750
14.2 (±1.1) 14.8 (±0.8) 14.5 (±1.3) 13.6 (± 2.2)
Ema et al. (1996c)
MBP
Rat (Wistar); PO, female (10-15/group)
0, 500, 625, 750 mg/kg-day
Gavage
GDs 13-15; dams sacrificed on GD 20
Number of implantations per litter (mean ± SD)
Dose
0
500
625
750
14.2 (±1.1) 14.4 (±2.4) 14.5 (±2.3) 14.2 (± 1.7)
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Ema and Mivawaki (2001)
MBP
Rat (Wistar); PO, female (16/group)
0, 250, 500, 750, 1,000 mg/kg-day
Gavage
GDs 0-8 with outcomes determined on
GD20
Ema et al. (2003)
MBzP
Rat (Wistar); PO, female (16/group)
0, 167, 250, 375 mg/kg-day
Gavage
GDs 15-17; dams sacrificed on GD 21
Saillenfait et al. (2003)
MBP
Rat (Sprague-Dawley); PO, female
(14-15/group)
0, 1.8, 3.6, 5.4 mmol/kg (equivalent to
0, 560, 1,120, 1,690 mg/kg as calculated
by study authors)
Gavage
GD 10; sacrificed on GD 21
Results by endpoint"
Number of corpora lutea per litter (mean ± SD)
Dose 0 250 500 750
16.5 16 (±1.2) 16.2 (±1) 16.4
(± 1.2) (± 1.8)
n = number of litters
1,000
15.9 (±0.9)
Number of implantations per female (mean ± SD)
Dose 0 250 500 750
15.5 14.6 14.6 13.2
(±1.3) (±2.5) (±4.2) (±5.4)
n = number of pregnant females
1,000
12.7
(±5.1)*
Number of implantations per litter (mean ± SD)
Dose 0 250 500 750
15.5 14.6 15.6 15.1
(±1.3) (±2.5) (±1.5) (±1.8)
n = number of litters
1,000
14.5 (± 1.3)
AGO
Data presented graphically; no significant effect on AGO
fetuses
of female
AGO per cube root of body weight ratio
Data presented graphically; no significant effect in female fetuses
Number of corpora lutea per litter (mean ± SD)
Dose 0 167 250
15.7 (±1.1) 15.1 (±1.3) 15.9 (±1.2)
375
16.1 (±1.1)
Number of implantations per litter (mean ± SD)
Dose 0 167 250
14.3 (±2) 13.5 (±1.5) 15.1 (±1.2)
375
14.8 (± 1.2)
Number of implantations per litter (mean ± SE)
Dose 0 560 1,120
13.73 14.62 13.75
(±0.73) (±0.63) (±0.68)
1,690
13.62 (±0.69)
Percent pregnant
Dose 0 560 1,120
79 93 86
Statistical significance not evaluated
1,690
87
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Saillenfait et al. (2003)
MBzP
Rat (Sprague-Dawley); PO, female
(12-14/group)
0, 0.9, 1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 280, 560, 1,120, 1,690 mg/kg as
calculated by study authors)
Gavage
GD 10; sacrificed on GD 21
Saillenfait et al. (2003)
MBP
Mouse (OF-1); PO, female (24-25/group)
0, 0.9, 1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 280, 560, 1,120, 1,690 mg/kg as
calculated by study authors)
Gavage
GD 8; sacrificed on GD 18
Saillenfait et al. (2003)
MBzP
Mouse (OF-1); PO, female (20-23/group)
0, 0.9, 1.8, 3.6, 5.4 mmol/kg (equivalent
to 0, 280, 560, 1,120, 1,690 mg/kg as
calculated by study authors)
Gavage
GD 8; sacrificed on GD 18
Male hormones
Shono et al. (2000)
MBP
Rat(Wistar-KingA)
Equivalent to 0 and 300 mg/kg-day
Gavage
CDs 15-18
Results by endpoint"
Number of implantations per litter (mean ± SE)
Dose 0 280 560 1,120
14 13.5 14.5 15.08
(±1.09) (±1.26) (±0.94) (±0.45)
Percent pregnant
Dose 0 280 560 1,120
93 92 86 93
Statistical significance not evaluated
Number of implantations per litter (mean ± SE)
Dose 0 280 560 1,120
13.45 13.71 11.27 12.73
(±0.89) (±0.65) (±1.04) (±0.72)
Percent pregnant
Dose 0 280 560 1,120
83 88 88 96
Statistical significance not evaluated
Number of implantations per litter (mean ± SE)
Dose 0 280 560 1,120
11.93 13.06 13.05 14.59
(±1.34) (±1.27) (±0.83) (±0.41)
Percent pregnant
Dose 0 280 560 1,120
71 80 83 86
Statistical significance not evaluated
Testosterone content of the testes (pg/testis,testis mean
Dose 0 300
852 (± 80.3) 50.9 (± 3.8)*
1,690
15 (±0.71)
1,690
75
1,690
13.24
(±0.75)
1,690
88
1,690
14.5
(±0.66)
1,690
86
±SE)
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Results by endpoint"
Male malformations
Shono et al. (2000)
MBP
Rat(Wistar-KingA)
Equivalent to 0 and 300 mg/kg-day
Gavage
CDs 15-18
Shono et al. (2000)
MBP
Rat(Wistar-KingA)
Equivalent to 0 and 300 mg/kg-day
Gavage
CDs 11-14
Shono et al. (2000)
MBP
Rat(Wistar-KingA)
Equivalent to 0 and 300 mg/kg-day
Gavage
CDs 7-10
Degree of transabdominal testicular migration (number of units from
bladder neck where 100 U = distance from bladder neck to lower pole
of kidney; mean ± SE)
Dose 0 300
9.3 (±1.9) 57.9 (±2.6)*
Epididymis: nonneoplastic lesions
Poorly developed epididymis
Testis: nonneoplastic lesions
No remarkable changes in the morphological features of Sertoli and
Leydig cells
Degree of transabdominal testicular migration (number of units from
bladder neck where 100 U = distance from bladder neck to lower pole
of kidney; mean ± SE)
Dose 0 300
9.3 (±1.9) 24.5 (±5.2)*
Degree of transabdominal testicular migration (number of units from
bladder neck where 100 U = distance from bladder neck to lower pole
of kidney; mean ± SE)
Dose 0 300
9.3 (±1.9) 12.3 (±5.9)
Male puberty, reproductive development
Ema et al. (2003)
MBzP
Rat (Wistar); PO, female (16/group)
0, 167, 250, 375 mg/kg-day
Gavage
GDs 15-17; dams sacrificed on GD 21
AGD
Data presented graphically; AGD significantly reduced at 250 and
375 mg/kg in male fetuses
AGD per cube root of body weight ratio
Data presented graphically; significantly lower in 250 and 375 mg/kg
groups than in control group
Degree of transabdominal testicular ascent (number of units from
bladder neck where 100 U = distance from bladder neck to lower pole
of kidney; mean ± SD)
Dose 0 167 250 375
18.9 (±0.3) 18.4 (±2.3) 23.8 (±7.1)* 40.1 (±8.2)*
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
Reference and study design
Shono and Suita (2003)
MBP
Rat (Wistar-King A); PO, female
(6/group)
0, 125, 250, 500, 1,000 mg/kg-day
Gavage
GDs 15-17; half of sacrificed on GD 20
for fetal examination; remaining
offspring examined PNDs 60-70
Results by endpoint"
Number of fetuses with undescended testes
Dose 0 167 250
2 1 21
375
79
Degree of transabdominal testicular ascent (number of units from
bladder neck where 100 U = distance from bladder neck to lower pole
of kidney; mean ± SD)
Dose 0 125 250 500
8.5 9.5 18.5 33.7
(±1.3) (± 1.4) (± 1.9)* (±2.8)*
1,000
58.6
(±2.1)*
Percent of fetuses with undescended testis
Dose 0 125 250 500
0 0 25* 61.1*
1,000
76.9*
1
2
3
4
5
6
*Result is statistically significant (p<0.05) based on analysis of data by study authors. * Results are presented as the
raw data as reported by the study authors.
- = for controls, no response relevant; for other doses, no quantitative response reported; (n) = number evaluated
from group; NR = not reported
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
l 3.4. PRELIMINARY MECHANISTIC INFORMATION FOR BBP
2 The systematic literature search for BBP also identified studies evaluating mechanisms of
3 action considered potentially relevant to effects observed following exposure to BBP. Studies were
4 included if they evaluated mechanistic events following exposure BBP or metabolites, or contained
5 information relevant to the mechanistic understanding of BBP toxicity. Reviews or analyses that do
6 not contain original data are not included here, but may be considered in later stages of assessment
7 development.
8 The diverse array of mechanistic studies presented here includes investigations of the
9 cellular, biochemical, and molecular mechanisms underlying toxicological outcomes. For this
10 preliminary evaluation, information reported in each study was extracted into a database (in the
11 form of an Excel spreadsheet) that will facilitate future evaluation of mechanistic information. This
12 information is being made available to provide an opportunity for stakeholder input, including the
13 identification of relevant studies not captured here.
14 The information extracted from each study and included in the database, corresponds to the
15 column headings in the spreadsheet, and is as follows: link to HERO record (contained within a URL
16 that links to the study abstract in the HERO database), HERO ID, author(s), year, molecular
17 formulation, in vitro/in vivo, species, cell type, endpoint(s) (i.e., mechanistic outcomes), assay, and
18 mechanistic category. The database supports sorting capabilities, e.g., data can be organized by
19 assay. The database is available through HERO at [http://hero.epa.gov/index.cfm?action=-
20 reference.details&reference id=2451132]. To access the database, click on the link at the top of the
21 web page and select "download" and then "ok" to view the spreadsheet in Excel. This spreadsheet
22 may also be saved to your desktop by downloading and selecting "save." The resulting inventory of
23 BBP mechanistic studies consists of 31 mechanistic outcomes from 18 in vivo studies, as well as
24 266 mechanistic outcomes from 84 in vitro assays. Table 3-35 presents a summary of the
25 mechanistic outcomes recorded in the database from each study identified.
26 The mechanistic categories developed here are not mutually exclusive and are designed to
27 facilitate the analysis of similar studies and experimental observations in a systematic manner.
28 This process will allow the identification of mechanistic events that contribute to mode(s) of action
29 (MOAs) and/or adverse outcome pathways (AOPs) following BBP exposure. The mechanistic
30 categories assigned to each mechanistic outcome reported by an individual study are as follows:
31 (1) mutation, including investigations of gene and chromosomal mutation; (2) DNA damage,
32 including indicator assays of genetic damage; (3) DNA repair; (4) oxidative stress; (5) cell death and
33 division (this captures a broad range of assays, but it is useful to consider them together as
34 observations resulting from cell cycle alterations; (6) pathology, which includes morphological
35 evaluations pertaining to the dysfunction of organs, tissues, and cells; (7) epigenetic effects, which
36 are observations of heritable changes in gene function that cannot be explained by changes in the
37 DNA sequence; (8) receptor-mediated and cell signaling effects; (9) immune system effects;
38 (10) cellular and molecular adsorption, distribution, metabolism, and excretion (ADME);
This document is a draft for review purposes only and does not constitute Agency policy.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
2
3
4
5
6
7
8
(11) cellular differentiation and transformation; (12) cellular energetics; and (13) "other," to
capture those mechanistic outcomes not easily assigned to a defined category. Mechanistic
outcomes in the "other" category include gene expression, proteomics and metabolomics arrays,
hormone production, and markers of angiogenesis. The ADME category above includes studies
reporting the cellular metabolism of BBP, thermodynamics of protein binding, and cellular
transport
Table 3-35. Summary of mechanistic outcomes evaluated following BBP
administration
Mechanistic
category
Mutation3
DNA damage
Total #
outcomes/
# studies
9/9
6/4
In vivo (#
outcomes/ft studies)
Total
0
1/1
Rat
0
0
Mouse
0
1/1
In vitro (# outcomes/ft studies)
Total
9/9
5/3
Human
0
1/1
Primate
0
0
Rat
0
0
Mouse
5/5
4/2
DNA repair
Oxidative stressb
Cell death and division0
Pathologyd
Epigenetics
Receptor-mediated
and cell signaling6
Immune system'
Cellular & molecular
ADME
Cellular differentiation
and transformation
Cellular energetics
Other8
Total
8/5
86/43
12/9
3/2
81/38
8/5
4/4
26/11
1/1
53/25
297/94
3/2
1/1
3/3
1/1
5/5
0
1/1
2/2
0
14/8
0
1/1
2/2
1/1
3/3
0
0
0
0
11/5
2/1
0
1/1
0
0
0
1/1
2/2
0
1/1
31/18
5/3
86/43
9/7
2/1
76/34
8/5
3/3
24/11
1/1
38/19
0
58/28
1/1
2/1
34/18
3/1
3/3
14/4
1/1
26/10
0
0
0
0
6/1
0
0
0
0
0
0
10/7
6/4
0
5/3
0
0
2/1
0
8/5
5/3
12/9
0
0
3/2
3/2
0
8/6
0
1/1
266/84
10
11
12
13
14
15
16
17
18
19
20
aDatabase included four outcomes in four studies utilizing Salmonella typhimuhum.
bDatabase included one outcome in one study utilizing fish in vivo.
°Database included one outcome in one study/each utilizing porcine or avian cells, and four endpoints from one
study of cultured bovine cells.
dDatabase included one outcome in one study/each utilizing porcine or hamster cells.
eDatabase included one outcome in one study/each utilizing fish or frogs in vivo; one endpoint from one study
using fish cells, two outcomes from one study/each using hamster or avian cells, five endpoints from two studies
using frog cells, and nine outcomes from five studies using bovine cells in vitro.
'Database included two outcomes in one study using rabbit cells in vitro.
This document is a draft for review purposes only and does not constitute Agency policy,
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1 gDatabase included two outcomes in one study/each using fish in vivo, as well as porcine, bovine or frog cells in
2 vitro; endpoints primarily consisted of gene expression, proteomics and metabolomics arrays, hormone
3 production, and markers of angiogenesis.
4
5 Notes: The number in rows may not sum to "total" amounts as several studies evaluated multiple species or
6 employed both in vivo and in vitro models. The mechanistic categories in italics and in gray shading had no BBP-
7 specific information available.
8
9 Information summarized in Table 3-35 and Figure 3-14 and detailed in the mechanistic
10 database can be used to ascertain the breadth and scope of available mechanistic studies. At this
11 preliminary stage, study results are not presented. Additionally, the inclusion of a study in the
12 spreadsheet does not reflect conclusions reached as to mechanistic study quality or relevance.
13 After the epidemiological and experimental studies on each health effect have been synthesized,
14 mechanistic studies will be reviewed and findings synthesized to evaluate potential MOAs and/or
15 AOPs, which can be used to inform hazard identification and dose-response assessment, specifically
16 addressing questions of human relevance, susceptibility, and dose-response relationships.
17
Mechanistic Outcomes
(297 outcomes from 94 reports)
DNA damage ^^|
OxiddTiye stress ^^|
Cell death and division ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^|
Irpigenetics |
Receptor-mediated and ceil signaling ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^|
18
19 Figure 3-22. Summary of in vivo or in vitro mechanistic data by mechanistic
20 category following oral exposure to BBP.
This document is a draft for review purposes only and does not constitute Agency policy.
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Preliminary Materials for the IRIS Toxicological Review of Butyl Benzyl Phthalate
1
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