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EPA Document# EPA-740-D-24-031
December 2024

United States	Office of Chemical Safety and

Environmental Protection Agency	Pollution Prevention

Draft Environmental Hazard Assessment for Butyl Benzyl

Phthalate (BBP)

Technical Support Document for the Draft Risk Evaluation

CASRN: 85-68-7

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28	TABLE OF CONTENTS	

29	ACKNOWLEGEMENTS	5

30	SUMMARY	6

31	1 INTRODUCTION	7

32	2 APPROACH AND METHODOLOGY	8

33	3 AQUATIC SPECIES HAZARD	10

34	4 TERRESTRIAL SPECIES HAZARD	17

35	5 ENVIRONMENTAL HAZARD THRESHOLDS	19

36	5.1 Aquatic Species COCs	19

37	5.2 Terrestrial Species Hazard Values	21

38	6 WEIGHT OF THE SCIENTIFIC EVIDENCE CONCLUSIONS FOR

3 9	ENVIRONMENTAL HAZARD	23

40	6.1 Strengths, Limitations, Assumptions, and Key Sources of Uncertainty for Environmental

41	Hazard	23

42	7 ENVIRONMENTAL HAZARD ASSESSMENT CONCLUSIONS	27

43	REFERENCES	28

44	Appendix A SPECIES SENSITIVITY DISTRIBUTION	34

45	Appendix B TERRESTRIAL VERTEBRATE TOXICITY OF BBP	37

46	Appendix C RUBRIC FOR WEIGHT OF THE SCIENTIFIC EVIDENCE	38

47	C.l Confidence Levels	38

48	C.2 Types of Uncertainties	38

49

50	LIST OF TABLES	

51	Table S-l Environmental Hazard Thresholds for BBP	6

52	Table 3-1. Acute Aquatic Vertebrate Toxicity of BBP	10

53	Table 3-2. Chronic Aquatic Vertebrate Toxicity of BBP	12

54	Table 3-3. Acute Aquatic Invertebrate Toxicity of BBP	13

55	Table 3-4. Chronic Aquatic Invertebrate Toxicity of BBP	14

56	Table 3-5. Aquatic Plant and Algae Toxicity of BBP	15

57	Table 4-1. Terrestrial Vertebrate Toxicity of BBP	17

58	Table 4-2. Terrestrial Invertebrate Toxicity of BBP	18

59	Table 4-3. Terrestrial Plant Toxicity of BBP	18

60	Table 5-1. Environmental Hazard Thresholds for BBP	22

61	Table 6-1. BBP Evidence Table Summarizing the Overall Confidence Derived from Hazard

62	Thresholds	26

63

64	LIST OF FIGURES	

65	Figure 5-1. Species Sensitivity Distribution (SSD) of Acute Hazard Effects of BBP on Aquatic

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line indicates the 5 percent Hazard Concentration (HC05 = 327 |ig/L)	20

LIST OF APPENDIX TABLES	

TableApx A-l. SSD Model Input for BBP Acute Exposure Toxicity in Aquatic Vertebrates and

Invertebrates - Empirical Data	34

Table Apx A-2. SSD Model Input for BBP Acute Exposure Toxicity in Aquatic Vertebrates and

Invertebrates - WeblCE Data	35

Table Apx A-3. SSD'' Model Predictions for Acute BBP Exposure Toxicity to Aquatic Vertebrates.... 36

Table_Apx B-l. Terrestrial Vertebrate Toxicity of BBP	37

TableApx C-l. Considerations that Inform Evaluations of the Strength of the Evidence within an

Evidence Stream (i.e., Apical Endpoints, Mechanistic, or Field Studies)	40

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ABBREVIATIONS AND ACRONYMS

AF

Assessment factor

BMD

Benchmark dose

BMDL

Benchmark dose limit

COC

Concentration(s) of concern

EC50

Effect concentration at which 50% of test organisms exhibit an effect

HC05

Hazard concentration that is protective of 95 percent of the species in the sensitivity



distribution

LC50

Concentration which is lethal to 50 percent of test organisms

LD50

Dose which is lethal to 50 percent of test organisms

LOAEL

Lowest-observable-adverse-effect4evel

LOEC

Lowest-observable-effect concentration

NAM

New approach method

NITE

National Institute of Technology and Evaluation

NOAEL

No-observed-adverse-effect level

NOEC

No-observed-effect concentration

NOEL

No-observed-effect level

OCSPP

Office of Chemical Safety and Pollution Prevention

OPPT

Office of Pollution Prevention and Toxics

PND

Postnatal day

POD

Point of departure

QSAR

Quantitative structure-activity relationship (model)

SSD

Species sensitivity distribution

TRV

Toxicity reference value

TSCA

Toxic Substances Control Act

U.S.

United States

Web-ICE

Web-based Interspecies Correlation Estimation

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ACKNOWLEGEMENTS	

This report was developed by the United States Environmental Protection Agency (U.S. EPA or the
Agency), Office of Chemical Safety and Pollution Prevention (OCSPP), Office of Pollution Prevention
and Toxics (OPPT).

Acknowledgements

The Assessment Team gratefully acknowledges the participation, review, and input from EPA OPPT
and OSCPP senior managers and science advisors. The Agency is also grateful for assistance from the
following EPA contractors for the preparation of this draft technical support document: General
Dynamics Information Technology, Inc. (Contract No. HHSN316201200013W); ICF, Inc. (Contract No.
68HERC23D0007); SpecPro Professional Services, LLC (Contract No. 68HERC20D0021); and SRC,
Inc. (Contract No. 68HERH19D0022 and 68HERC23D0007).

As part of an intra-agency review, this technical support document was provided to multiple EPA
Program Offices for review. Comments were submitted by EPA's Office of Research and Development
(ORD).

Docket

Supporting information can be found in the public docket, Docket ID EPA-HQ-QPPT-2018-0501.
Disclaimer

Reference herein to any specific commercial products, process or service by trade name, trademark,
manufacturer, or otherwise does not constitute or imply its endorsement, recommendation, or favoring
by the United States Government.

Authors: Collin Beachum (Management Lead), Brandall Ingle-Carlson (Assessment Lead), Randall
Bernot (Environmental Hazard Assessment Lead), Jennifer Brennan, Christopher Green (Environmental
Hazard Discipline Leads)

Contributors: Azah Abdallah Mohamed, Rony Arauz Melendez, Sarah Au, Maggie Clark, Jone
Corrales, Daniel DePasquale, Lauren Gates, Emily Griffin, Ryan Klein, Sydney Nguyen, Brianne
Raccor, Maxwell Sail, Joe Valdez, Leora Vegosen.

Technical Support: Hillary Hollinger, S. Xiah Kragie

This report was reviewed by OPPT and OCSPP leadership.

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SUMMARY	

This technical document is in support of the TSCA Draft Risk Evaluation for butyl benzyl phthalate
(BBP) (U.S. EPA. 2025). BBP is a common chemical name for the chemical substance 1,2-
benzenedicarboxylic acid, 1-butyl 2-(phenylmethyl) ester (CASRN 85-68-7).

EPA considered all reasonably available information identified through the systematic review process
under the Toxic Substances Control Act (TSCA) to characterize environmental hazard endpoints for
BBP. After evaluating the reasonably available information, environmental hazard thresholds were
derived for aquatic vertebrates, aquatic invertebrates, aquatic plants and algae, and terrestrial vertebrates
(Table S-l).

Table S-l Environmental Hazard Thresholds for BBP

Receptor Group

Exposure
Duration

Hazard Threshold (COC
or HV)

Citation

Aquatic Vertebrates

Acute

197 |ig/L

From SSD; See Section 5

Chronic

1.9 |ig/L

(Battelle, 2018c)

Aquatic Invertebrates

Acute

197|ig/L

From SSD; See Section 5

Chronic

62.6 |ig/L

(Rhodes et al.. 1995)

Aquatic Plants and Algae

Chronic

21 |ig/L

(Adams et al., 1995)

Terrestrial Vertebrates

Chronic

311 mg/kg/day

(TNO. 1993)

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159	1 INTRODUCTION	

160	Butyl benzyl phthalate is a clear, oily liquid with a total production volume in the United States between

161	10 and 50 million pounds (U.S. EPA. 2020). Butyl benzyl phthalate is manufactured (including

162	imported) in the United States. The chemical is processed as a reactant, incorporated into a formulation,

163	mixture, or reaction product, and incorporated into articles.

164

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2 APPROACH AND METHODOLOGY	

TSCA requires that EPA use data and/or information in a manner consistent with the best available
science and that EPA base decisions on the weight of scientific evidence. To meet the TSCA science
standards, EPA applies a systematic review process to identify data and information across taxonomic
groups for both aquatic and terrestrial organisms with a focus on apical endpoints (e.g., those affecting
survival, growth, or reproduction). The data collection, data evaluation, and data integration stages of
the systematic review process are used to develop the hazard assessment to support the integrative risk
characterization. EPA uses several considerations when weighing and weighting the scientific evidence
to determine confidence in the environmental hazard data. These considerations include the quality of
the database, consistency, strength and precision, biological gradient/dose response, and relevance. EPA
completed the review of environmental hazard data/information sources during risk evaluation using the
data quality review evaluation metrics and the rating criteria described in the 2021 Draft Systematic
Review Protocol supporting TSCA Risk Evaluations for Chemical Substances (U.S. EPA. 2021) and
Draft Risk Evaluation for Butyl Benzyl Phthalate (BBP) - Systematic Review Protocol (U.S. EPA.
2024c). Studies identified and evaluated by OPPT through 2020 were assigned an overall quality level
of high, medium, low, or uninformative. Data on toxicity of BBP are numerous and, in some instances,
vary substantially, thus EPA systematically evaluated all data for this hazard characterization, but relied
upon only high-quality and medium-quality studies for purposes of quantitative risk characterization.
References receiving an overall quality determination of low or uninformative either exceeded the BBP
limit of solubility in all treatments, showed no effects at the highest concentration tested, evaluated a
biotransformation (mechanistic) endpoint, and/or were part of a mixture.

EPA reviewed potential environmental hazards associated with BBP. EPA considered all available
studies to characterize the environmental hazards of BBP to surrogate species representing various
receptor groups, including aquatic vertebrates, aquatic invertebrates, amphibians, aquatic plants, algae,
and birds. Mechanistic (transcriptomic and metabolomic) and behavioral points of departure from one
study of an acute exposure of BBP to fathead minnows were used to inform of the potential mechanisms
that lead to the acute and chronic aquatic vertebrate hazard thresholds (Bencic et al.. 2024). Hazard
studies with mammalian wildlife exposed to BBP were not available, therefore EPA used ecologically
relevant endpoints from human health laboratory rat and mouse model organisms to establish a hazard
threshold for terrestrial mammals.

A Species Sensitivity Distribution (SSD) analysis was used to derive an acute aquatic hazard threshold.
An SSD is a model of the variation in sensitivity of species to a particular chemical stressor and is
generated by fitting a statistical distribution function to the proportion of species affected as a function
of concentration or dose. Empirical data that were included in the SSD analysis were limited to LC50
values (concentration which is lethal to 50% of test organisms) that were at or below the limit of water
solubility of 2690 |ig/L for BBP (U.S. EPA. 2024a). Specifically, predicted hazard data were generated
using EPA's Web-Based Interspecies Correlation Estimation Web-ICE (v4.0) toxicity predictions tool
(Raimondo. 2010). The species and corresponding empirical data are outlined in Section 5 and
Appendix A. EPA derived concentrations of concern (COC) for all other organism and exposure
durations using studies that report hazard effects at or below the limit of water solubility of 2690 |ig/L
g/L for BBP.

Environmental Hazard from Previous Assessments

Environment Canada previously assessed environmental hazard effects of BBP (EC. 2000). Through a
survey of acute exposure (48-hour and 96-hour durations) studies of organism mortality that estimated
concentrations which are lethal to 50% of test organisms (LC50s), aquatic acute hazard was determined

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213	to be 510 |ig/L for the shiner perch (Cymcitogcister aggregata). Aquatic chronic exposure hazards and

214	algal exposure hazards were not identified (EC. 2000). The European Union (EU) Risk Assessment

215	Report (ECJRC. 2007) reports the lowest acute aquatic hazard value as 510 |ig/L BBP for C. aggregata

216	(ECJRC. 2007). The EU assessment also reports the lowest chronic NOEC (No-observed-effect

217	concentration) values as 140 |ig/L BBP to fish (30-day exposure to Pimephalespromelas), 75 |ig/L BBP

218	to an invertebrate (28-day exposure to Americamysis bahia), and 200 |ig/L BBP to a diatom (72-hour

219	exposure to Naviculapelliculosa) (ECJRC. 2007). Neither assessment reports hazard threshold data on

220	the effects of BBP to terrestrial organisms.

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3 AQUATIC SPECIES HAZARD	

EPA reviewed 51 studies for BBP toxicity to aquatic organisms. Some studies may have included
multiple endpoints, species, and test durations. Four of these studies received an overall quality
determination of low, uninformative, or did not meet systematic review criteria. The data from these low
or uninformative studies were not used to derive hazard thresholds because they either exceeded the
BBP limit of solubility in all treatments, showed no effects at the highest concentration tested, evaluated
a biotransformation (mechanistic) endpoint, and/or were part of a mixture. Forty-seven studies received
an overall quality determination high or medium quality, were used to derive hazard thresholds, and are
detailed in the subsections below. Studies that demonstrated no acute or chronic adverse effects at the
highest concentration tested (unbounded NOECs), or where hazard values exceeded the limit of
solubility for DBP in water as determined by EPA at 2690 |ig/L, (U.S. EPA, 2024, 11799672) are
included in Table 3-1, Table 3-2, Table 3-3, Table 3-4, and Table 3-5, but were excluded from
consideration for the development of hazard thresholds (Section 5). Additionally, predicted hazard data
for 18 species were generated using EPA's Web-ICE (v4.0) tool (Raimondo, 2010). including
predictions for 14 fish, and four invertebrate species. No toxicity studies using spiked sediment for
benthic exposures were identified for BBP. Thus, all hazard data to benthic invertebrates were
represented by water exposures.

Acute Aquatic Vertebrates

EPA reviewed seven high/medium quality studies for acute toxicity in aquatic vertebrates (Table 3-1).
Of these studies, six contained acceptable endpoints that identified definitive hazard values below the
BBP limit of water solubility (2690 |ig/L). For the fathead minnow (Pimephales promelas), bluegill
{Lepomis macrochirus), rainbow trout (Oncorynchus mykiss), and shiner perch (Cymatogaster
aggregcita) the 96-hour mortality LC50s ranged from 510 to 2100 |ig/L BBP (Adams et al.. 1995;
Ozretich et al.. 1983; EG&G Bionomics. 1979a. c, d). These values were combined with acute hazard
effects values of BBP to aquatic invertebrates to derive an SSD and subsequent acute exposure threshold
(Appendix A).

Table 3-1. Acute Aquatic Vertebrate Toxicity of BBP

Test Organism

Hazard Values

Duration

Endpoint

Citation
(Study Quality)

Fathead minnow

{Pimephales

promelas)

1500 |ig/L a

96-hour LC50

Mortality

(Adams et al., 1995)
(High)

2100 |ig/L a

96-hour LC50

Mortality

(EG&G Bionomics,
1979d)(High)

Bluegill

{Lepomis
macrochirus)

1700 |ig/L a

96-hour LC50

Mortality

(EG&G Bionomics,
1979c) (Medium)

Sheepshead
minnow

{Cyprinodon
variegatus)

3000 |ig/L b

96-hour NOEC

Mortality

(EG&G Bionomics,
1979a) (Medium)

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Test Organism

Hazard Values

Duration

Endpoint

Citation
(Study Quality)

Rainbow trout

(Oncorynchus
mykiss)

820 |ig/L a

96-hour LC50

Mortality

(Ozretich et al.,
1983)(High)

3300 |ig/L b

96-hour LC50

Mortality

(EG&G Bionomics,
1979d)(High)

Shiner perch

(Cymatogaster
aggregcita)

510 |ig/L a

96-hour LC50

Mortality

(Ozretich et al.,
1983) (Medium)

11 Value used as input for SSD derivation of acute aquatic hazard threshold.
h Hazard value is greater than the BBP limit of solubility (2690 ng/L).

TSCA section 4(h)(1)(B) requires EPA to encourage and facilitate the use of scientifically valid test
methods and strategies that reduce or replace the use of vertebrate animals while providing information
of equivalent or better scientific quality and relevance that will support regulatory decisions. In line with
EPA's New Approach Methods Work Plan, EPA OPPT and ORD have been collaborating on
developing new methods for use in TSCA risk evaluations. Specifically, a project was conducted to
generate omics-based PODs and compared them to traditional endpoints using fathead minnow as the
model organism for three of the phthalates undergoing a TSCA risk evaluation, including BBP (Bencic
et al.. 2024). In this study, points of departure (PODs) were derived for transcriptomic change (tPOD; 60
|ig/L), metabolomic change (mPOD; 120 |ig/L), and behavioral change (bPOD 90 |ig/L) resulting from
24-hour duration of aquatic BBP exposure to fathead minnows. Additionally, a 24-hour mortality
NOEC/LOEC of 1000 /2000 |ig/L was identified. In 2000 |ig/L BBP exposures, 38 percent mortality
was observed. These results suggest that fathead minnow larvae exhibited changes in gene expression,
metabolite levels, and swimming behavior at sublethal concentrations of BBP. While hazard thresholds
are usually calculated with in vivo data measuring an apical endpoint (e.g., mortality, reproduction,
growth), these mechanistic (transcriptomic and metabolomic) and behavior points of departure represent
potential information that may be used for reducing the time needed for toxicity testing in vivo and
provide an alternate method to characterize hazard as well as provide important evidence for
mechanisms of action. At this time, EPA has not used the omics-based PODs in the BBP draft risk
evaluation. There are uncertainties with respect to the extent to which these sub-organismal and
individual-level effects (e.g., behavior) at short exposure durations are comparable to ecologically
relevant outcomes, such as survival and reproduction, in wild fish populations.

Chronic Aquatic Vertebrates

EPA reviewed eight high or medium quality studies for chronic toxicity in aquatic vertebrates (Table
3-2). Of these studies, four contained acceptable chronic endpoints that identified definitive hazard
values below the BBP limit of water solubility (2690 |ig/L), for four fish species. One study found
effects of BBP on amphibian growth (Battelle, 2018a). Another study of dietary BBP exposure to the
fish, Sander lucioperca, found slightly reduced growth and female skewed sex ratios after five weeks of
high doses (360 g/kg bw/day) of BBP amended diets (Jarmolowicz et al.. 2014). However, feeding
treatments were not replicated and diet concentrations were not verified analytically.

Chronic water exposure studies include a 21-day reproduction test of BBP exposure to zebrafish (Danio
rerio), which found 3% lower fecundity, 2% lower fertilization success, 100% increase in plasma
vitellogenin, and reduced gonad weight in males in treatments with 33 |ig/L BBP (Lowest-observable-

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effect concentration, LOEC) (Battelle, 2018c). No effects were observed at 11 pg/L BBP (NOEC).

In a separate study, fewer (10% less) eggs per Japanese medaka (Oryzias latipes) female were found
after five weeks of exposure to 95 pg/L BBP, but no effects on fertilization rates, growth, gonad weight,
or plasma vitellogenin were found in the same study (Battelle, 2018b). Other chronic exposure studies
resulted in no growth or reproductive effects of BBP to rainbow trout (Oncorhynchus mykiss) (Rhodes et
al.. 1995) or fathead minnow {Pimephalespromelas) (ABC Laboratories. 2008) (Table 3-2). Fish
behaviors may also be altered due to chronic BBP exposure, as Mummichog (Fundulus heteroclitus)
shoaled with smaller fish when exposed for 28-days to 100 pg/L BBP compared to control fish that
shoaled with larger fish (Kaplan et al.. 2013).

Table 3-2. Chronic Aquatic Vertebrate Toxicity of BBP

Test Organism

Hazard Values

Duration

Endpoint

Citation
(Study Quality)

African clawed
frog (Xenopus
laevis)

No hazard effects;
Greater growth in
all BBP exposures

21-day
LOEC

Growth

(Battelle, 2018a)
(High)

Zebrafish

(Danio rerio)

11/33 ng/L°

21-day

NOEC/LOEC

Reproduction

(Battelle, 2018c)
(High)

Rainbow trout

(Oncorhynchus
mykiss)

>200 pg/L
No effects
observed

21-day

Mortality and
Growth

(Rhodes et al.. 1995)
(High)

Japanese
medaka

{Oryzias
latipes)

35/95 pg/L b

5-week
NOEC/LOEC

Growth

(10% reduction in
egg production)

(Battelle. 2018b)
(Medium)

Fathead
minnow
(Pimephales
promelas)

>65 pg/L

164-day
NOEC

Growth and
Reproduction

(ABC Laboratories,
2008)(High)

> 82 pg/L

6-week

Reproduction

(ABC Laboratories,
2008)(High)

Mummichog

(Fimduhis
heteroclitus)

100 pg/L

28-day
LOEC

Behavior

(Kaplan et al., 2013)
(High)

European
pikeperch

{Sander
lucioperca)

180.0/360.0 g/kg

bw/day

NOEC/LOEC

5-week diet
exposure

Reproduction and
Growth

(Jarmolowicz et al.,
2014) (Medium)

a 3% lower fecundity; 2% lower fertilization success; 100% increase in plasma vitellogenin; reduced gonad
weight in males.

h 10% fewer eggs per female; no effects on fertilization rates, growth, gonad weight, or plasma vitellogenin.
Bolded number indicates the values used to derive the chronic exposure Concentration of Concern (COC).

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Acute Aquatic Invertebrates

EPA reviewed 17 high or medium quality studies for acute toxicity in aquatic invertebrates (Table 3-3).
Fifty percent mortality effects (LC50s) or short-term effects (EC50s) of acute exposures of BBP to
aquatic invertebrates ranged from 0.46 mg/L to concentrations of BBP above the limit of water
solubility (i.e., >2690 |ig/L). Of these studies, seven contained acceptable endpoints that identified
definitive hazard values below the BBP limit of water solubility (2690 |ig/L). These values were
combined with acute hazard effects values of BBP to aquatic invertebrates to derive an SSD and
subsequent acute exposure threshold (Appendix A). For midge (Chironomus teutons), amphipod
(Hyalella azteca), mayfly, (Hexagenia sp.) opossum shrimp (Americamysis bahia), Taiwan abalone
(Haliotis diversicolor), and Virginia oyster (Crassostrea virginica), acute BBP water exposure resulted
in LC50 values ranging from 460 |ig/L to 2650 |ig/L BBP.

Table 3-3. Acute Aquatic Invertebrate Toxicity of BBP

Test Organism

Hazard
Values

Duration

Endpoint

Citation
(Study Quality)

Midge

(Chironomus tentans)

1640 |ig/L a

48-hour
LC50 (no
sediment)

Mortality

(Monsanto, 1982)
(Medium)

3600 |ig/L b

48-hour LC50

Mortality

(SRI International
1984) (Medium)

Amphipod

(Hyalella azteca)

460 |ig/L a

10-day
LC50 (no
sediment)

Mortality

(Call etal.. 2001a)
(High)

Mayfly

(Hexagenia sp.)

1100 |ig/L a

96-hour LC50

Mortality

(ABC Laboratories,
1986c) (High)

Opossum shrimp

(Americamysis bahia)

1100 |ig/L a

96-hour LC50

Mortality

(Springborn
Bionomics, 1988)
(High)

900 |ig/L a

96-hour LC50

Mortality

(EG&G Bionomics,
1979b)(High)

Moina macrocopa
(Water Flea)

3690 |ig/L b

48-hour LC50

Immobilization

(Wang et al„ 2011)
(High)

Crayfish

(Procambarus sp.)

>2400 |ig/L

96-hour LC50

Mortality

(ABC Laboratories,
1986b) (high)

(Polychaete worm)
(Nereis virens)

> 3000 |ig/L b

96-hour LC50

Mortality

(Springborn
Bionomics, 1986b)
(High)

Taiwan abalone
(Haliotis diversicolor)

2650 |ig/L a

96-hour EC50

Growth

(Liu et al., 2009)
(High)

Virginia oyster
(Crassostrea virginica)

1300 |ig/L a

96-hour EC50

Growth

(ABC Laboratories,
1986a) (High)

Hydra

(Hydra littoralis)

>1920 |ig/L

96-hour LC50

Mortality

(ABC Laboratories,
1986a) (High)

Pink shrimp

(Penaeus duorarum)

>3400 |ig/L

96-hour LC50

Mortality

(Soringborn
Bionomics, 1986a)
(High)

Midge

(Paratanytarsus

>3600 |ig/L

48-hour LC50

Mortality

(SRI International,
1984) (Medium)

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Test Organism

Hazard
Values

Duration

Endpoint

Citation
(Study Quality)

dissimilis)









Midge

(Paratanytarsus
parthenogenetica)

7200 |ig/L b

48-hour LC50

Mortality

(Monsanto, 1983 a)
(High)

Waterflea

(Daphnia magna)

>1400 |ig/L

48-hour LC50

Immobilization

(SDringborn
Bionomics, 1984)
(Medium)

>960 |ig/L

48-hour LC50

Immobilization

(Adams et al., 1995)
(High)

a Value used as input for SSD derivation of acute aquatic hazard threshold.
h Hazard value is greater than the BBP limit of solubility (2690 ng/L).

Chronic Aquatic Invertebrates

EPA reviewed six high or medium quality studies for chronic toxicity in aquatic invertebrates (Table
3-4). All six studies contained acceptable chronic endpoints that identified definitive hazard values
below the BBP limit of water solubility (2690 |ig/L). Chronic effects of BBP on aquatic invertebrates
ranged from reduced opossum shrimp (Americamysis bahia) reproduction after 28-days at 170 |ig/L
BBP (Springborn Bionomics, 1986c) to growth reduction in midges (Chironomus tentcms) after 10-days
at 1420 |ig/L BBP (Call etal.. 2001bV

In a 21-day study of Daphnia magna, 80% mortality and 70% fewer offspring per female occurred when
exposed to 1400 |ig/L BBP compared to no-BBP control treatments (Rhodes et al.. 1995). Daphnia
magna exposed to BBP in a 21-day static renewal bioassay produced 50% fewer offspring at 220 |ig/L
BBP (LOEC) but were not affected at 350 |ig/L BBP (NOEC) (Monsanto. 1983b). In a study that lasted
42-days, 35% fewer/), magna survived in 760 |ig/L BBP compared to control treatments (EG&G
Bionomics. 1979e).

Rotifer (Brachionus calyciflorus) population growth rates were also reduced in chronic BBP exposures
(Cruciani et al.. 2015; Zhao et al.. 2009). In a 96-hour exposure experiment, B. calyciflorus population
growth rates were reduced by 25% at 2000 |ig/L (Cruciani et al.. 2015). In another study with a 144-
hour chronic exposure duration, B. calyciflorus population growth rates were reduced by 15% at 500
|ig/L BBP (Zhao et al.. 2009). In a 28-day exposure experiment, Americamysis bahia reproductive
success (offspring/female/day) was reduced by 50% when exposed to 170 |ig/L BBP (Springborn
Bionomics. 1986c). In a 10-day water exposure experiment, the oligochaete worm (Lumbricuius
variegatus) survival was reduced by 50% when exposed to 1230 |ig/L BBP (Call et al.. 2001b). In a 10-
day water exposure experiment, the midge (Chironomus tentcms) dry weight was reduced by 50% when
exposed to 1420 |ig/L BBP (Call et al.. 2001b).

Table 3-4. Chronic Aquatic Invertebrate Toxicity of BBP

Test Organism

Hazard Values

Duration

Endpoint

Citation
(Study Quality)

Rotifer

(Brachionus
calyciflorus)

1000/2000 |ig/L
NOEC/LOEC

96-hour

Population growth
rate

(Cruciani et al.,
2015) (Medium)

50/500 |ig/L
NOEC/LOEC

144-hour

Population growth
rate

(Zhao et al., 2009)
(Medium)

Waterflea

280/1400 jig/L

21-day

Mortality

(Rhodes et al., 1995)

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Test Organism

Hazard Values

Duration

Endpoint

Citation
(Study Quality)

(Daphnia magna)

NOEC/LOEC





(High)

4800 |ig/L

160-hour EC50

Immobilization

(Monsanto, 1983 c)
(Medium)

220/350 |ig/L
NOEC/LOEC

21-day

Reproduction

(Monsanto, 1983b)
(Medium)

260/760 |ig/L
NOEC/LOEC

Two generation
(42-day)

Mortality

(EG&G Bionomics,
1979e) (High)

Opossum shrimp

(Americamysis bahia)

75/170 |ig/L
(NOEC/LOEC)

28-day

Reproduction

(Springborn
Bionomics, 1986c)
(High)

Oligochaete worm
(Lumbriculus
variegatus)

1230 |ig/L

10-day

(no sediment)

Mortality

(Call et al.. 2001b)
(High)

Midge

(Chironomus tentans)

1420 |ig/L

10-day EC50
(no sediment)

Growth

(Call et al.. 2001b)
(High)

Bolded number indicates the values used to derive the chronic exposure Concentration of Concern (COC).

Aquatic Plants and Algae

EPA reviewed nine high or medium quality studies for toxicity in aquatic plants and algae (Table 3-5).
Eight of these studies found population level hazard effects (96-h EC50) that ranged from 210 |ig/L
(green algae Raphidocelis subcapitata) to 600 |ig/L (diatoms Naviculapelliculosa and Skeletonema
costatum) and were less than the BBP limit of water solubility (2690 |ig/L) (Adams et al.. 1995; EG&G
Bionomics. 1978). A study of the cyanobacterium, Microcystis aeruginosa, did not find effects of BBP
on population growth rate (EG&G Bionomics. 1978). Cyanobacterium are bacteria and not algae or
plants, but EPA includes this study to illustrate the differential types of effects of BBP on different taxa
(U.S. EPA. 2021Y

Table 3-5. Aquatic Plant and Algae Toxicity of BBP

Test Organism

Hazard Values

Duration

Endpoint

Citation
(Study Quality)

Raphidocelis
subcapitata
(Green Algae)

210 jig/L

96-hour EC50

Population

(Adams et al., 1995)
(High)

400 |ig/L

96-hour EC50

Population

(EG&G Bionomics,
1978) (Medium)

Navicula pelliculosa
(Diatom)

600 |ig/L

96-hour EC50

Population

(EG&G Bionomics,
1978) (Medium)

410 |ig/L

72-hour E50

Population

(Carolina Ecotox,
1995a) (High)

Skeletonema costatum
(Diatom)

600 |ig/L

96-hour EC50

Population

(EG&G Bionomics,
1978) (Medium)

Dunaliella tertiolecta
(Green Algae)

1000 |ig/L

96-hour EC50

Population

(EG&G Bionomics,
1978) (Medium)

Microcystis
aeruginosa (Blue-
Green Algae)"

>1000000 |ig/L

96-hour EC50

Population

(EG&G Bionomics,
1978) (Medium)

Scenedesmus

330 |ig/L

72-hour EC50

Population

(Carolina Ecotox,

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Test Organism

Hazard Values

Duration

Endpoint

Citation
(Study Quality)

subspicatus (Green
algae)







1995b)(High)

Chlorella vulgaris
(Green Algae)

>2880 |ig/L

72-hour EC50

Population

(Carolina Ecotox, 1997)
(High)

11 Cyanobacterial species, not algae.

Bolded number indicates the values used to derive the algal Concentration of Concern (COC).

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4 TERRESTRIAL SPECIES HAZARD	

EPA assigned an overall quality level of high or medium to five acceptable studies containing hazard
data for seven different taxa. These studies contained relevant toxicity data for the Norway rat (Rattus
norvegicus), the chicken (Gallus gallus), the nematode (Caenorhabditis elegans), and four plant species
(Ipomoea aquatica, Trifolium repens, Sinapis alba, Brassica rapa).

Terrestrial Vertebrates

No reasonably available information was identified for exposures of BBP to wild mammalian
populations. In lieu of wild mammal studies, EPA reviewed nine studies on BBP hazard to laboratory
rodents that were designed to determine human health hazards of BBP that also contained ecologically
relevant reproductive endpoints (Table Apx B-l). Thus, EPA used data from laboratory rodent studies
as surrogates for the potential BBP hazards to wild mammal populations. EPA's decision to focus on
ecologically relevant (population level) reproductive endpoints in the rat and mouse data set for BBP for
consideration of a hazard threshold in terrestrial mammals is due to the sensitivity of these taxa to BBP
in eliciting phthalate syndrome (U.S. EPA, 2024b). Of the nine rat and mouse studies containing
ecologically relevant reproductive endpoints, EPA selected the study with the most sensitive LOAEL
(lowest observed adverse effect level) for evaluating data quality and for deriving the hazard threshold
for terrestrial mammals. The most sensitive reproductive endpoint was from a study that involved the
Sprague-Dawley strain of Norway rat (Rattus norvegicus) (TNO. 1993). with a 136-day LOAEL of 446
mg/kg-bw/day BBP and NOAEL (no observed adverse effect level) of 217 mg/kg-bw/day for reduced
pup weight. This study was assigned an overall quality determination of high. This study found
significantly decreased pup weights (males, females, and combined) on postnatal day (PND) 21 in the
second litter only (no effect in first litter) at 446 mg/kg-bw/day. Males were exposed for 10 weeks pre-
mating, during mating and until sacrifice on day 161. Exposure to F0 females was for 2 weeks pre-
mating, during mating (up to 3 weeks), gestation (~3 weeks) and lactation (~3 weeks) of litter FOa, for 7-
13 days after weaning (1-2 weeks), and during mating (up to 3 weeks), gestation (~3 weeks) and
lactation (~3 weeks) of litter FOb. The female premating mean dose was used for the NOAEL and
LOAEL because it is the lowest mean dose value for females across premating, gestation, and lactation.

One study of BBP effects on chicken (Gallus gallus) hens administered 5 g/kg bw/day BBP to birds on
days 1 to 3 and again on days 21 to 23 of a 42-day experiment (University of Arizona. 1978). Hens fed
this regime of BBP laid >90% fewer eggs over the course of 42 days compared to control hens. This
study exposed hens to BBP at only one dose; therefore, EC50s were not derived. Also, oral doses were
administered directly but by unknown methods and BBP doses were not analytically verified.

Table 4-1. Terresi

trial Vertebrate Toxicity of BBP

Test Organism

Hazard Values

Duration

Endpoint

Citation
(Study Quality)

Norway rat

(Rattus

norvegicus)

217 mg/kg bw/d
NOAEL and 446 mg/kg
bw/d LOAEL
311 mg/kg bw/d
geometric mean of
NOAEL and LOAEL

136 days

Reduced pup
weight during
lactation;
increased pup
mortality at PND
2-4

(TNO. 1993)
(High)

Chicken (Gallus
gallus)

5 g/kg bw/d

BBP added to
diet on days 1 to
3 and days 21 to
23 of 42 day

Reproduction;
>90% fewer eggs
produced in one
treatment dose

(Universitv of
Arizona, 1978)
(Medium)

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experiment





Terrestrial Invertebrates

EPA reviewed one medium quality study for BBP toxicity in a terrestrial invertebrate (Table 4-2). The
study exposed the soil nematode Caenorhabditis elegcms to water solutions of BBP. No nematode
mortality after 24-hours occurred up to and including 100,000 |ig/L BBP (Kwon et al.. 2011). Also, the
exposure concentration of 100,000 |ig/L is well above the limit of water solubility for BBP (2690 |ig/L
(U.S. EPA. 2024b)). indicating that these experimental conditions are unlikely to occur in ecosystems.

Table 4-2. Terrestrial Invertebrate Toxicity of BBP

Test Organism

Hazard Values

Duration

Endpoint

Citation
(Study Quality)

Nematode

{Caenorhabditis
elegans)

>100,000 |ig/L
NOEC

24-hour

Mortality

(Kwon et al.,
2011) (Medium)

Terrestrial Plants

EPA reviewed four high or medium quality studies for BBP toxicity in terrestrial plants (Table 4-3). A
study of Ipomoea aquatica (Swamp Morning glory) found a 50% reduction in plant biomass after 21-
day s of hydroponic water exposure to 100,000 |ig/L BBP (LOEC), but plant biomass was not affected
when exposed to 50,000 |ig/L BBP (Chen et al.. 2011). The exposure concentration of 100,000 |ig/L is
well above the limit of water solubility for BBP (2690 |ig/L (U.S. EPA. 2024b)). indicating that these
experimental conditions are unlikely to occur in ecosystems. One study exposed three plant species to
BBP vapor over 21-days. No BBP vapor-phase concentration affected plant growth to Trifolium repens
(Dutch Clover), Sinapis alba (White Mustard), Brassica rapa (Bird Rape) (Gorsuch et al.. 2008).

Table 4-3. Terrestrial Plant Toxicity of BBP

Test Organism

Hazard
Values

Duration

Endpoint

Citation
(Study Quality)

Ipomoea aquatica
(Swamp morning
glory)

50,000 |ig/L
NOEC and
100,000 |ig/L
LOEC

28-day

Growth

(Chen et al., 2011) (Hish)

Trifolium repens
(Dutch clover)

>5.7 |ig/m3
NOEL

21-day

Vapor-phase
toxicity

(Gorsuch et al., 2008)
(High)

Sinapis alba (White
mustard)

>5.7 |ig/m3
NOEL

21-day

Vapor-phase
toxicity

(Gorsuch et al., 2008)
(High)

Brassica rapa (Bird
rape,)

>5.7 |ig/m3
NOEL

21-day

Vapor-phase
toxicity

(Gorsuch et al., 2008)
(High)

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5 ENVIRONMENTAL HAZARD THRESHOLDS	

EPA calculates hazard thresholds to identify potential concerns to aquatic and terrestrial species. After
weighing the scientific evidence, EPA selects the appropriate toxicity value from the integrated data to
use for hazard thresholds. Table 5-1 summarizes the concentrations of concern (COCs) identified for
BBP. See Section 6 for more details about how EPA weighed the scientific evidence.

In aquatic species, EPA uses probabilistic approaches (e.g., SSD) when data from at least eight species
(Raimondo. 2010) are available and deterministic approaches (e.g., deriving a geometric mean of several
comparable values) when limited data are available. For BBP, an SSD was derived for acute aquatic
exposure hazards and a deterministic approach was used to assess chronic hazard in aquatic and
terrestrial taxa. For the deterministic approaches, COCs are calculated by dividing a hazard value by an
assessment factor (AF) according to EPA methods (U.S. EPA. 2016. 2013. 2012).

Equation 5-1

COC = toxicity value '¦ AF

For terrestrial species, EPA estimates hazard by calculating a toxicity reference value (TRV) or by
assigning the hazard threshold as the most sensitive and ecologically relevant reproductive endpoint in
the case of mammals, birds, and terrestrial plants.

5.1 Aquatic Species COCs	

Acute Aquatic Concentration of Concern

For aquatic species, EPA uses probabilistic approaches (e.g., SSD) when data from at least eight species
(Raimondo. 2010) data are available. An SSD is a model of the variation in sensitivity of species to a
particular chemical stressor and is generated by fitting a statistical distribution function to the proportion
of species affected as a function of concentration or dose. It can be used to visualize which species are
most sensitive to a toxic chemical exposure, and to predict the concentration of a toxic chemical that is
hazardous to a percentage of test species. This hazardous concentration (HC) is represented as an HCp,
where p is the percent of species below the threshold. EPA used an HC05 (a hazardous concentration
threshold for 5% of species) to estimate a concentration that is protective of 95% of species. This HC05
can then be used to derive a COC, and the lower bound of the 95th percent confidence interval (CI) of
the HC05 can be used to account for uncertainty instead of dividing by an AF. EPA has more confidence
in the probabilistic approach compared to the deterministic approach when enough data are available
because an HC05 is representative of a larger proportion of species in the environment.

The aquatic acute COC for BBP was derived from an SSD that contained LC50s for five fish species
and six invertebrate species identified in systematic review, bolstered by an additional 18 predicted
LC50 values from the Web-ICE v4.0 toxicity value estimation tool. Web-ICE is a tool developed by
U.S. EPA's Office of Research and Development that estimates the acute toxicity of a chemical to a
species, genus, or family from the known toxicity of the chemical to a surrogate species. It was used to
obtain estimated acute toxicity values for BBP in species that were not represented in the empirical data
set. (Figure 5-1). SSDs were derived using EPA's SSD Toolbox (vl.l) (Etterson. 2020) and plotted
using R Statistical Software (v4.4.1) (R Core Team. 2019) using the ssdtools R package (vl.0.6) and the
ggplot2 R package (v3.5.1; Appendix A). All studies included in the SSD were rated high or medium
quality. The Maximum Likelihood method and a Weibull distribution model were used. The Weibull
distribution was based on an examination of Akaike's Information Criterion corrected for sample size

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(aicc) for goodness of fit ( kirnham and Anderson. 2002). visual examination of Q-Q plots, and
evaluation of the line of best fit near the low-end of the SSD. The HC05 for this distribution was 327
|ig/L BBP with a 95% confidence interval of 197 ug/L to 552 |ig/L. After taking the lower 95th percent
confidence interval of this HC05 as an alternative to the use of assessment factors, the acute aquatic
COC for vertebrates and invertebrates was 197 jig/L BBP (Figure 5-1).

Species Sensitivity for BBP

100%

80° o

T3


a 60%'
to
<
in
11

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to 626 |ig/L (626 |ig/L) and applied an AF of 10, resulting in a COC = 62.6 jig/L BBP.

Aquatic Algae Concentration of Concern

Of the eight studies that investigated the effects of BBP on algae, EPA derived a COC based on the
lowest and most protective EC50 value which was 210 |ig/L for BBP hazard effects on the green algae
Raphidocelis subcapitata. EPA calculated a COC by applying an AF of 10, resulting in a COC = 21
ji«/L BBP.

5.2 Terrestrial Species Hazard Values

Terrestrial Mamnuil Hazard Threshold

Nine laboratory rat and mouse studies were assessed with the most sensitive and ecologically relevant
reproductive endpoint value chosen to represent the terrestrial mammalian hazard threshold. Phthalates
were filtered to identify those with reproductive effects as the most sensitive endpoints. The terrestrial
mammalian hazard threshold was derived from the most sensitive among acceptable-quality studies
involving the Sprague-Dawley strain of Norway rat (Rattus norvegicus) (TNO. 1993). with a 136-day
LOAEL of 446 mg/kg-bw/day BBP and NOAEL of 217 mg/kg-bw/day for reduced pup weight. EPA
calculated a geometric mean of the NOAEL and LOAEL from this study to equal the hazard threshold
of 311 mg/kg-bw/day BBP.

Avian Hazard Threshold

One study of BBP effects on chicken (Gallus gallus) hens administered 5 g/kg bw/day BBP to birds on
days 1 to 3 and again on days 21 to 23 of a 42-day experiment (University of Arizona. 1978). Hens fed
this regime of BBP laid >90% fewer eggs over the course of 42 days compared to control hens. This
study exposed BBP to hens at only one dose; therefore, EC50s via a dose-response experimental design
could not be derived. Also, oral doses were administered directly but by unknown methods. The
methods do not describe if or how BBP was added to food rations or any methods for analytically
verifying BBP doses. No other evidence of BBP toxicity to birds was reasonably available to consider
for a hazard threshold. EPA did not derive an avian hazard threshold due to these uncertainties in
experimental design and analysis from the one available study.

Terrestrial Invertebrate Hazard Threshold

EPA reviewed one medium quality study for BBP toxicity in a terrestrial invertebrate (Table 4-2). The
study exposed the soil nematode Ccienorhcibditis elegcms to water solutions of BBP. No nematode
mortality after 24 hours occurred up to and including 100,000 |ig/L BBP (Kwon et al.. 2011). No other
evidence of BBP toxicity to terrestrial invertebrates was reasonably available to consider for a hazard
threshold. Thus, EPA did not derive a terrestrial invertebrate hazard threshold.

Terrestrial Plants Hazard Threshold

EPA reviewed four high or medium quality studies for BBP toxicity in terrestrial plants (Table 4-3). A
study of Ipomoea aquatica (Swamp Morning glory) found a 50% reduction in plant biomass after 21
days of hydroponic exposure to 100,000 |ig/L BBP (LOEC), but plant biomass was not affected when
exposed to 50000 |ig/L BBP (Chen et al.. 2011). This study exposed plants to water well above the BBP
limit of water solubility (2690 |ig/L) in a hydroponic scenario. Other available studies exposed plants to
BBP fumigant and found no hazard effects up to and including the highest concentrations of exposure.
No other evidence of BBP toxicity to terrestrial plants in soil was reasonably available to consider for a
hazard threshold. Thus, EPA did not derive a terrestrial plant hazard threshold.

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534

535	Table 5-1. Environmental Hazard Thresholds for BBP

Receptor Group

Exposure
Duration

Hazard Threshold (COC
or HV)

Citation

Aquatic Vertebrates

Acute

197 |ig/L

From SSD

Chronic

1.9 |ig/L

(Battelle, 2018c)

Aquatic Invertebrates

Acute

197 |ig/L

From SSD

Chronic

62.6 |ig/L

(Rhodes et al.. 1995)

Aquatic Plants and Algae

Chronic

21 |ig/L

(Adams et al., 1995)

Terrestrial Vertebrates

Chronic

311 mg/kg/day

(TNO. 1993)

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6 WEIGHT OF THE SCIENTIFIC EVIDENCE CONCLUSIONS FOR
ENVIRONMENTAL HAZARD	

EPA uses several considerations when weighing and weighting the scientific evidence to determine
confidence in the environmental hazard data. These considerations include the quality of the database,
consistency, strength and precision, biological gradient/dose response, and relevance. This approach is
described in the Draft Systematic Review Protocol Supporting TSCA Risk Evaluations for Chemical
Substances (U.S. EPA. 2021). Table 6-1 summarizes how these considerations were determined for each
environmental hazard threshold. Criteria for assessing confidence is described in Appendix C Evidence
Integration.

EPA determined that BBP poses hazards from acute and chronic exposures to aquatic vertebrates, acute
and chronic exposures to aquatic invertebrates, chronic exposure to algae, and chronic dietary exposure
to terrestrial mammals. EPA has robust confidence in the weight of evidence in these findings.

6.1 Strengths, Limitations, Assumptions, and Key Sources of Uncertainty
for Environmental Hazard

The weight of evidence suggests that BBP poses acute hazard effects to vertebrate and invertebrate
animals at 197 jug/L BBP. EPA has robust confidence in this hazard threshold because the quality of the
database of studies included 11 high or medium quality studies that consistently resulted in LC50s
between 460 |ig/L (Lake Superior Research Institute. 1997) up to 2650 |ig/L BBP (Liu et al.. 2009).
These studies all were conducted with reasonable dose-response designs and results, which enabled
precise LC50 calculations (Table 3-1 and Table 3-3). These hazard effects were documented across a
range of species that live in freshwater and marine environments in the water column as well as in or
near the benthos/sediment. Additional consideration of acute (24-hour) larval fish transcriptomics,
metabolomics, and behavior data revealed within-organism effects occurring in the same order of
magnitude (ranging from 60 |ig/L to 120 |ig/L BBP), consistent with the hypothesis that hazard occurs
at similar exposures. EPA used a probabilistic technique (SSD) to derive a COC that is protective of
95% of the aquatic animals in a community by incorporating hazard values across species and habitats.
Limitations of SSDs include its reliance on model species that may not exist or interact in the same
ecological community and are weighted equally. Another assumption that may limit the scope of SSD
inference is whether the number of species used is adequate. The shape of the data distribution that is
fitted to the effects data can be subjective and dependent on the three or four lowest values (Newman et
al.. 2000). Notwithstanding the limitations of SSD analyses, this method is widely used and accepted in
risk assessments. Thus, EPA has robust confidence in the quality, consistency, strength and precision,
and relevance of the studies used in determining the acute aquatic COC (197 jug/L BBP).

The weight of evidence suggests that BBP poses chronic hazard effects to vertebrate animals at 1.9 jug/L
BBP. EPA has robust confidence in the hazard threshold for four reasons. First, the reasonably available
database of studies used for this determination includes eight high or medium quality studies to
determine growth or reproduction effects using standard methods. Second, these studies were conducted
on a range of different species including zebrafish (Danio rerio), fathead minnow (Pimephales
promelas), and Japanese medaka (Oryzias latipes) (Table 3-2). Third, these studies found consistent
effects within the same order of magnitude of BBP concentrations. Finally, all of these studies were
conducted with reasonable dose-response designs and results, which enabled precise estimations of
effect concentrations. Thus, EPA has robust confidence in the quality, consistency, strength and
precision, and relevance of the studies used in determining the chronic aquatic COC for vertebrates (1.9
ji«/L BBP).

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The weight of evidence suggests that BBP poses chronic hazard effects to invertebrate animals at 62.6
ji«/L BBP. EPA has robust confidence in the hazard threshold for four reasons. First, the reasonably
available database of studies used for this determination includes six high or medium quality studies to
determine growth or reproduction effects using standard methods. Second, these studies were conducted
on a range of different species including rotifers (Brachionus calyciflorus), water fleas {Daphnia
magna), opossum shrimps (Americamysis bahia), oligochaete worms (Lumbricuius variegatus), and
midges (Chironomus tentans), representing three different phyla (Table 3-4). Third, these studies found
consistent effects within the same order of magnitude of BBP concentrations. Finally, all of these studies
were conducted with reasonable dose-response designs and results, which enabled precise estimations of
effect concentrations. Thus, EPA has robust confidence in the quality, consistency, strength and
precision, and relevance of the studies used in determining the chronic aquatic COC for invertebrates
(62.6 jig/L BBP)

The weight of evidence suggests that BBP poses chronic hazard effects to algae at 21 jug/L BBP. EPA
has robust confidence in the hazard threshold for four reasons. First, the reasonably available database of
studies used for this determination includes eight high or medium quality studies to determine
population growth effects of BBP using standard methods. Second, these studies were conducted on a
range of different species including green algae (Raphidocelis subcapitata, Dunaliella tertiolecta,
Scenedesmus subspicatus, and Chlorella vulgaris) and diatoms (Naviculapellicidosa and Skeletonema
costatum) representing two different phyla (Table 3-5). Third, these studies found consistent effects
within the same order of magnitude of BBP concentrations. Finally, all of these studies were conducted
with reasonable dose-response designs and results, which enabled precise estimations of effect
concentrations. Thus, EPA has robust confidence in the quality, consistency, strength and precision, and
relevance of the studies used in determining the chronic aquatic COC for algae (21 jug/L BBP).

No studies on terrestrial wildlife involving mammals were identified. In lieu of terrestrial wildlife
studies, nine references for rat studies as human health model organisms were used to determine a
lowest and most conservative BBP concentration that affected apical endpoints (survival, reproduction,
growth) in rodents and that could serve as an indication of hazard effects in wild mammal populations.
The weight of evidence suggests that BBP poses chronic dietary exposure hazard effects to terrestrial
mammals at 311 mg/kg bw/day BBP. EPA has robust confidence in this hazard threshold for three
reasons (Table 6-1). First, the reasonably available database of studies used for this determination
include nine high or medium quality studies to determine reproductive effects of BBP using standard
methods. The terrestrial mammalian hazard threshold was derived from the most sensitive among
acceptable-quality studies involving the Sprague-Dawley rat (Rattus norvegicus) (TNO. 1993). with a
136-day LOAEL of 446 mg/kg-bw/day BBP and NOAEL of 217 mg/kg-bw/day for reduced pup weight.
Second, these nine studies found consistent effects within the same order of magnitude of BBP doses.
Finally, all of the studies were conducted with reasonable dose-response designs and results, which
enabled precise estimation of effect concentrations. However, ecologically relevant population level
effects were not observed in ecologically relevant species. Considerable uncertainties surround whether
or how these effects on individual growth and reproductive development translate into effects on wild
mammal fitness and population parameters. Because of these uncertainties of extrapolations to wildlife
mammal species, EPA has moderate confidence that the hazards are representative of the range of wild
mammal species. Therefore, EPA has robust confidence in the quality, consistency, and strength and
precision, of the studies used in determining the hazard threshold for terrestrial mammals (311 mg/kg
bw/day BBP), but moderate confidence in their relevance to wild mammal populations.

EPA has less confidence in the use of one avian study (University of Arizona. 1978). one terrestrial

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632	invertebrate study (Kwon et al.. 2011). and one terrestrial plant study (Chen et al.. 2011) to derive

633	hazard thresholds for these groups for many reasons. First, as only one study is available for each taxa,

634	consistency across studies is unknown. Second, each study has at least one limitation in study design or

635	analysis that limits the precision, biological gradient/dose response, and/or relevance of their results. For

636	example, the study of C. elegans worms and the study of plant Ipomoeci aquatica (Swamp Morning

637	glory) exposed organisms to concentrations (100000 |ig/L in both cases) well above the limit of

638	solubility of BBP (2690 |ig/L). The study of BBP effects on chicken egg production had limited

639	descriptions of the methods and of dose administration and analytical verification (University of

640	Arizona. 1978). Therefore, EPA has slight confidence in the quality, consistency, strength and precision,

641	and relevance of these studies and did not derive hazard thresholds for these organisms.

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Table 6-1. BBP Evidence Table Summarizing the Overall Confidence Derived from Hazard Thresholds

Types of Evidence

Quality of
the Database

„ . ^ Strength and
Consistency L

Precision

Biological

Gradient/Dose-

Response

Relevance

Hazard
Confidence

Aquatic

Acute aquatic assessment

+++

+++

+++

+++

+++

Robust

Chronic aquatic assessment

+++

+++

+++

+++

+++

Robust

Algal assessment

+++

+++

+++

+++

+++

Robust

Terrestrial

Chronic mammalian assessment

+++

+++

+++

+++

++

Robust

Chronic avian assessment





+

+

++

Slight

Terrestrial invertebrate assessment





+

+

++

Slight

Terrestrial plant assessment





+

+

++

Slight

3 Relevance includes biological, physical/chemical, and environmental relevance.

+++ Robust confidence suggests thorough understanding of the scientific evidence and uncertainties. The supporting weight of scientific evidence
outweighs the uncertainties to the point where it is unlikely that the uncertainties could have a significant effect on the hazard estimate.

++ Moderate confidence suggests some understanding of the scientific evidence and uncertainties. The supporting scientific evidence weighed against
the uncertainties is reasonably adequate to characterize hazard estimates.

+ Slight confidence is assigned when the weight of scientific evidence may not be adequate to characterize the scenario, and when the assessor is
making the best scientific assessment possible in the absence of complete information. There are additional uncertainties that may need to be considered.


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7 ENVIRONMENTAL HAZARD ASSESSMENT CONCLUSIONS

EPA considered the quality, consistency, strength and precision, biological gradient/dose response, and
relevance of the reasonably available data to weigh the scientific evidence in determining the
environmental hazards of BBP. EPA determined that BBP poses acute and chronic exposure hazards to
aquatic vertebrates, acute and chronic exposure hazards to aquatic invertebrates, chronic exposure
hazards to algae, and chronic dietary exposure hazards to terrestrial mammals. BBP hazards include:

Aquatic species

•	LC50 values from 11 acute duration exposures of BBP to aquatic fish and invertebrates were
used to develop an SSD. The lower 95% confidence value of the HC05 was used as the COC at
197 |ig/L BBP.

•	The most sensitive aquatic vertebrate for which a clear population4evel fitness endpoint could be
obtained was for the zebrafish (Danio rerio). This 21-day reproduction test of BBP exposure to
IX rerio found 3% lower fecundity, 2% lower fertilization success, 100% increase in plasma
vitellogenin, and reduced gonad weight in males in treatments with 33 |ig/L BBP (LOEC). No
effects were observed at 11 |ig/L BBP (NOEC). Based on the presence of a clear dose-response
relationship and a population-level fitness endpoint, the 21-day ChV for reduction in
reproduction was selected to derive the chronic COC for aquatic vertebrates as 1.9 |ig/L BBP.

•	A 21-day study of Daphnia magna found 80% mortality and 70% fewer offspring per female due
to BBP chronic exposure, leading to a COC of 62.6 |ig/L BBP for chronic invertebrate hazard.

•	EPA derived a COC for chronic algal BBP exposure from the EC50 value of 210 |ig/L to the
green algae Raphidocelis sabcapitata resulting in a COC of 21 |ig/L BBP.

Terrestrial Species

•	The terrestrial mammalian hazard threshold was derived from the most sensitive among
acceptable-quality studies involving the Sprague-Dawley rat (Rattus norvegicus) with a 136-day
dietary exposure hazard threshold of 311 mg/kg-bw/day BBP.

•	No evidence of BBP toxicity to terrestrial invertebrates was reasonably available to consider for
a hazard threshold. Thus, EPA did not derive a terrestrial invertebrate hazard threshold.

•	No evidence of BBP toxicity to terrestrial plants in soil was reasonably available to consider for
a hazard threshold. Thus, EPA did not derive a terrestrial plant hazard threshold.


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091586. (252.1085.6114.522. BW-86-7-2094. OTS0522399. 40-8626256. 42005 B5-7.
TSCA/406078). Monsanto Co.

https://ntrl.ntis. gov/NTRL/dashboard/searchResults.xhtml?searchQuery=OTS0522399
Springborn Bionomics. (1986c). Chronic toxicity of butylbenzyl phthalate to mysid shrimp (Mysidopsis
bahia) [Bionomics Report #BW-86-7-2074] [TSCA Submission], In Four acute toxicity studies,
one chronic toxicity study and an uptake on an elimination study with 14C-benzyl butyl
phthalate with attachments and cover letter dated 091586. (252-1085-6113-530. BW-86-7-2074.
OTS0522399. 40-8626256. 42005 B5-7. TSCA/406078). St. Louis, MO: Monsanto Co.
https://ntrl.ntis. gov/NTRL/dashboard/searchResults.xhtml?searchOuerv=OTSQ522399
Springborn Bionomics. (1986d). Six acute and chronic toxicity reports regarding butylbenzyl phthalate.

(40-8626222). St. Louis, MO: Monsanto Chemical Co.

Springborn Bionomics. (1988). Acute toxicity of benzyl butyl phthalate to mysid shrimp (Mysidopsis
bahia) under flow-through conditions with cover letter dated 011888 [TSCA Submission],
(252.0687.6118.510/SLS# 87-10-2525. OTS0522497. 40-8826335. 42005 I2B-2.
TSCATS/406178). Monsanto Company.

https://ntrl.ntis.gov/NTRL/dashboard/searchResults/titleDetail/OTS0522497.xhtml
SRI International. (1984). Acute toxicity studies on S-160 using two midge species as the test organisms

[TSCA Submission], (EPA/OTS Doc #878213936). St. Louis, MO: Monsanto.

Streufort. JM. (1978). Some effects of two phthalic acid esters on the life cycle of the midge
(Chironomus plumosus) [TSCA Submission], (OTS0000013-0. FYI-AX-1178-0013.
TSC ATS/029296). Washington, DC: Manufacturing Chemists Association.
https://ntrl.ntis.gov/NTRL/dashboard/searchResults/titleDetail/OTS0000013Q.xhtml
TNO. (1993). Dietary one-generation reproduction study with butyl benzyl phthalate in rats with cover
letter dated 040793 [TSCA Submission], (EPA/OTS Doc #86-930000189). St. Louis, MO:
Monsanto Co.

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https://ntrl.ntis. gov/NTRL/dashboard/searchResults.xhtml?searchOuerv=OTSQ538169
TNO. (1998). [Redacted] Oral developmental reproduction study with butyl benzyl phthalate in Wistar

rats. (V98.408 final). European Council for Plasticizers and Intermediates.

U.S. EPA. (1998). Guidelines for ecological risk assessment [EPA Report], (EPA/630/R-95/002F).
Washington, DC: U.S. Environmental Protection Agency, Risk Assessment Forum.
https://www.epa.gov/risk/guidelines-ecological-risk-assessment
U.S. EPA. (2005). Guidelines for carcinogen risk assessment [EPA Report], (EPA630P03001F).
Washington, DC. https://www.epa.gov/sites/production/files/2013-
09/documents/cancer guidelines final 3-25-05.pdf
U.S. EPA. (2012). Sustainable futures: P2 framework manual [EPA Report], (EPA/748/B-12/001).
Washington DC. http://www.epa.gov/sustainable-futures/sustainable-futures-p2-framework-
manual

U.S. EPA. (2013). Interpretive assistance document for assessment of discrete organic chemicals.
Sustainable futures summary assessment [EPA Report], Washington, DC.
http://www.epa.gov/sites/production/files/2015-05/documents/05-iad discretes iune2013.pdf
U.S. EPA. (2016). Weight of evidence in ecological assessment [EPA Report], (EPA/100/R-16/001).
Washington, DC: Office of the Science Advisor.
https://nepis.epa. gov/Exe/ZvPURL.cgi?Dockev=P100SFXR.txt
U.S. EPA. (2020). Final scope of the risk evaluation for butyl benzyl phthalate (1,2-benzenedicarboxylic
acid, 1-butyl 2-(phenylmethyl) ester); CASRN 85-68-7 [EPA Report], (EPA-740-R-20-015).
Washington, DC: Office of Chemical Safety and Pollution Prevention.
https://www.epa.gov/sites/default/files/2020-09/documents/casrn 85-68-
7 butyl benzyl phthalate finalscope.pdf
U.S. EPA. (2021). Draft systematic review protocol supporting TSCA risk evaluations for chemical
substances, Version 1.0: A generic TSCA systematic review protocol with chemical-specific
methodologies. (EPA Document #EPA-D-20-031). Washington, DC: Office of Chemical Safety
and Pollution Prevention. https://www.regulations.gov/document/EPA-HQ-OPPT-2021-0414-
0005

U.S. EPA. (2024a). Draft Physical Chemistry and Fate and Transport Assessment for Butyl Benzyl

Phthalate (BBP). Washington, DC: Office of Pollution Prevention and Toxics.

U.S. EPA. (2024b). Draft Physical Chemistry Assessment for Butyl benzyl phthalate (BBP).

Washington, DC: Office of Pollution Prevention and Toxics.

U.S. EPA. (2024c). Draft Systematic Protocol for Butyl Benzyl Phthalate (BBP). Washington, DC:

Office of Pollution Prevention and Toxics.

U.S. EPA. (2025). Draft Risk Evaluation for Butyl Benzyl Phthalate (BBP). Washington, DC: Office of

Pollution Prevention and Toxics.

University of Arizona. (1978). Initial submission: Evaluation of butyl benzyl phthalate with laying hens
with cover letter dated 080792 [TSCA Submission], (EPA/OTS Doc #88-920007247). St. Louis,
MO: Monsanto Co.

https://ntrl.ntis.gov/NTRL/dashboard/searchResults/titleDetail/OTS0545565.xhtml
Wang. JX; Xi. YL; Flu. K; Liu. XB. (2011). Effect of butyl benzyl phthalate on life table-demography of

two successive generations of cladoceran Moina macrocopa Straus. J Environ Biol 32: 17-22.
Wolf. C: Lambright. C: Mann. P; Price. M; Cooper. RL; Ostbv. J: Gray. LE. Jr. (1999). Administration
of potentially antiandrogenic pesticides (procymidone, linuron, iprodione, chlozolinate, p,p'-
DDE, and ketoconazole) and toxic substances (dibutyl- and diethylhexyl phthalate, PCB 169,
and ethane dimethane sulphonate) during sexual differentiation produces diverse profiles of
reproductive malformations in the male rat. Toxicol Ind Health 15: 94-118.
http://dx.doi.org/10.1177/0748233799015001Q9
Zhao. LL; Xi. YL: Huang. L; Zha. CW. (2009). Effects of three phthalate esters on the life-table

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951	demography of freshwater rotifer Brachionus calyciflorus Pallas. Aquatic Ecology 43: 395-402.

952	http://dx.doi.org/10.1007/slQ452-008-9179-6

953

954

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Appendix A SPECIES SENSITIVITY DISTRIBUTION	

An SSD was derived using only acute duration exposure studies that calculated LC50s. The SSD
Toolbox is a resource that can fit SSDs to environmental hazard data (Etterson. 2020). It runs on Matlab
2018b (9.5) for Windows 64 bit. For this draft BBP risk evaluation, EPA created one SSD with the SSD
Toolbox Version 1.1 to evaluate acute aquatic vertebrate and invertebrate toxicity. The use of this
probabilistic approach increases confidence in the hazard threshold identification as it is a more data-
driven way of accounting for uncertainty. For the acute SSD, acute exposure hazard data for aquatic
vertebrates and invertebrates were curated to prioritize study quality and to assure comparability
between toxicity values. For example, the empirical data set included only LC50s for high and medium
quality acute duration assays that measured mortality for aquatic vertebrates and invertebrates.
TableApx A-l shows the empirical data and TableApx A-2 shows the modelled data from Web-ice
that were used in the SSD.

With this data set, the SSD Toolbox was used to apply a variety of algorithms to fit and visualize SSDs
with different distributions. An HC05 was calculated for each. The SSD Toolbox's output contained
several methods for choosing an appropriate distribution and fitting method, including goodness-of-fit,
standard error, and sample-size corrected Akaike Information Criterion (AICc, (Burnham and Anderson,
2002)). Most p-values for goodness-of-fit were less than 0.05, showing no evidence of lack of fit. The
distribution and model with the lowest AICc value, and therefore the best fit for the data was the
Weibull Distribution (Table_Apx A-3). Because numerical methods may lack statistical power for small
sample sizes, a visual inspection of the data were also used to assess goodness-of-fit. For the Q-Q plot,
the horizontal axis gives the empirical quantiles while the vertical axis gives the predicted quantiles
(from the fitted distribution). The Q-Q plot demonstrates a good model fit with the data points in close
proximity to the line across the data distribution. Q-Q plots were visually used to assess the goodness-
of-fit for the distributions with the Weibull distribution demonstrating the best fit near the low end of the
distribution, which is the region from which the HC05 is derived. The results for this model (Figure
5-1) predicted 5 percent of the species (HC05) to have their LC50s exceeded at 377 |ig/L (154 to 531
|ig/L 95% CI).

Table Apx A-l. SSD Model Input for BBP Acute Exposure Toxicity in Aquatic Vertebrates and
Invertebrates - Empirical Data			

Species

Description

Acute Toxicity Value LC50
(Hg/L)

Citation(s)

Hycdella ctztecct

Aquatic
invertebrate

460

(Lake Superior Research
Institute. 1997; Adams et al..
1995: EG&G Bionomics. 1984)

Cymatogaster aggregate!

Aquatic
vertebrate

510

(Chen et al.. 2014; Ozretich et
al.. 1983)

Oncorhvnchiis mykiss

Aquatic
vertebrate

820

(Ozretich et al.. 1983)

Americamvsis bahia

Aquatic
invertebrate

1100

(EG&G Bionomics. 1979b)

900

(Snrinaborn Bionomics. 1988)

Hexagenia sp.

Aquatic
invertebrate

1100

(Adams et al.. 1995;
EnviroSvstem. 1991; ABC
Laboratories. 1986c; EG&G
Bionomics. 1983)

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Species

Description

Acute Toxicity Value LC50
(Hg/L)

Citation(s)

Crcissostreci virginica

Aquatic
invertebrate

1300

(ABC Laboratories. 1986a;
Linden etal.. 1979)

Chironomns tentans

Aquatic
invertebrate

1640

(Monsanto. 1982)

Lepomis macrochirns

Aquatic
vertebrate

1700

(EG&G Bionomics. 1979c;
Streufort. 1978)

Pimephcdes promelas

Aquatic
vertebrate

1500

(Adams et al.. 1995)

2100

(EG&G Bionomics. 1979d)

Haliotis diversicolor

Aquatic
invertebrate

2650

(Liu et al.. 2009)

986

987	TableApx A-2. SSD Model Input for BBP Acute Exposure Toxicity in Aquatic Vertebrates and

988	Invertebrates - WeblCE Data

Species

Description

Acute Toxicity Value LC50
(^g/L)

Caecidotea brevicauda

Invertebrate

447

Gammarus pseudolimnaeus

Invertebrate

480

Ceriodaphnia dubia

Invertebrate

523

Salve linus namaycush

Fish

637

Oncorhynchus clarkii

Fish

702

Perca flavescens

Fish

715

Oncorhynchus kisutch

Fish

766

Salmo trutta

Fish

851

Salmo solar

Fish

937

Oncorhynchus tshawytscha

Fish

965

Micropterus salmoides

Fish

1022

Poecilia reticulata

Fish

1306

Cyprinus carpio

Fish

1902

Cyprinodon variegatus

Fish

1915

Ictalurus punctatus

Fish

1916

Daphnia magna

Invertebrate

1919

Carassius auratus

Fish

2315

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989

990	Table Apx A-3. SSD" Model Predictions for Acute BBP Exposure Toxicity to Aquatic Vertebrates

Distribution6

HC05 (jig/L)

p- value

Weibull

327

0.93

Normal

475

0.70

Logistic

467

0.66

Gumbel

487

0.38

Burr

464

0.63

11 The SSD was generated using SSD Toolbox vl.l.

h The model with the lowest AICc value, and therefore the best model fit, is bolded in this table.

991

992

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Appendix B TERRESTRIAL VERTEBRATE TOXICITY OF BBP

In lieu of wild mammal studies, EPA considered nine studies on BBP to laboratory rodents that were
designed to determine human health hazards of BBP that also contained ecologically relevant
reproductive endpoints (TableApx B-l). Of the studies containing ecologically relevant reproductive
endpoints to rat and mouse, EPA selected the study with the most sensitive LOAEL (lowest observed
adverse effect level) for evaluating data quality and for deriving the hazard threshold for terrestrial
mammals (Table Apx B-l).

Table Apx B-l. Terrestrial Vertebrate Toxicity of

BBP

Test Organism
(Species)

Hazard Values

Duration

Endpoint

Citation

Rat (Rattus
norvegicus)

250/500 mg/kg-
bw/day

GD 15 - 17

Reproduction

(Ema and Mivawaki.
2002)

500/750 mg/kg-
bw/day

GD 5 - 17

(Ema et al.. 1992)

247/821 mg/kg-
bw/day

Two

generation

(SDrineborn Bionomics.
1986d; Nikonorow et al..
1973)

500/1000 mg/kg-
bw/day

29 days

(Wolf et al.. 1999;
Piersma et al.. 1995)

419/1641 mg/kg-
bw/day

GD 6 - 15

(RTI International. 1989)

254/2270 mg/kg-
bw/day

10 weeks

(Hazelton Labs. 1985)

0.115/0.321
mg/kg-bw/day

9 weeks
drinking water

(TNO. 1998)

Mice

247/821 mg/kg-
bw/day

Two

generation

(NTP. 1990)

910/2330 mg/kg-
bw/day

GD 6 - 15

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Appendix C RUBRIC FOR WEIGHT OF THE SCIENTIFIC
	EVIDENCE	

The weight of the scientific evidence fundamentally means that the evidence is weighed (i.e., ranked)
and weighted (i.e., a piece or set of evidence or uncertainty may have more importance or influence in
the result than another). Based on the weight of the scientific evidence and uncertainties, a confidence
statement was developed that qualitatively ranks (i.e., robust, moderate, slight, or indeterminate) the
confidence in the hazard threshold. The qualitative confidence levels are described below.

The evidence considerations and criteria detailed within U.S. EPA (2021) guides the application of
strength-of-evidence judgments for environmental hazard effect within a given evidence stream and
were adapted from Table 7-10 of the 2021 Draft Systematic Review Protocol (U.S. EPA. 2021).

EPA used the strength-of-evidence and uncertainties from U.S. EPA (2021) for the hazard assessment to
qualitatively rank the overall confidence rating for environmental hazard (Table Apx C-l). Confidence
levels of robust (+ + +), moderate (+ +), slight (+), or indeterminant are assigned for each evidence
property that corresponds to the evidence considerations (U.S. EPA. 2021). The rank of the Quality of
the Database consideration is based on the systematic review overall quality determination (High,
Medium, or Low) for studies used to calculate the hazard threshold, and whether there are data gaps in
the toxicity data set. Another consideration in the Quality of the Database is the risk of bias (i.e., how
representative is the study to ecologically relevant endpoints). Additionally, because of the importance
of the studies used for deriving hazard thresholds, the Quality of the Database consideration may have
greater weight than the other individual considerations. The high, medium, and low systematic review
overall quality determinations ranks correspond to the evidence table ranks of robust (+ + +), moderate
(+ +), or slight (+), respectively. The evidence considerations are weighted based on professional
judgment to obtain the overall confidence for each hazard threshold. In other words, the weights of each
evidence property relative to the other properties are dependent on the specifics of the weight of the
scientific evidence and uncertainties that are described in the narrative and may or may not be equal.
Therefore, the overall score is not necessarily a mean or defaulted to the lowest score. The confidence
levels and uncertainty type examples are described below.

C.l Confidence Levels	

•	Robust (+ + +) confidence suggests thorough understanding of the scientific evidence and
uncertainties. The supporting weight of the scientific evidence outweighs the uncertainties to the
point where it is unlikely that the uncertainties could have a significant effect on the exposure or
hazard estimate.

•	Moderate (+ +) confidence suggests some understanding of the scientific evidence and
uncertainties. The supporting scientific evidence weighed against the uncertainties is reasonably
adequate to characterize exposure or hazard estimates.

•	Slight (+) confidence is assigned when the weight of the scientific evidence may not be adequate
to characterize the scenario, and when the assessor is making the best scientific assessment
possible in the absence of complete information. There are additional uncertainties that may need
to be considered.

C.2 Types of Uncertainties	

The following uncertainties may be relevant to one or more of the weight of the scientific evidence
considerations listed above and will be integrated into that property's rank in the evidence table:

•	Scenario Uncertainty: Uncertainty regarding missing or incomplete information needed to fully

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define the exposure and dose.

o The sources of scenario uncertainty include descriptive errors, aggregation errors, errors
in professional judgment, and incomplete analysis.

•	Parameter Uncertainty: Uncertainty regarding some parameter.

o Sources of parameter uncertainty include measurement errors, sampling errors,
variability, and use of generic or surrogate data.

•	Model Uncertainty: Uncertainty regarding gaps in scientific theory required to make predictions
on the basis of causal inferences.

o Modeling assumptions may be simplified representations of reality.

Table 6-1 summarizes the weight of the scientific evidence and uncertainties, while increasing
transparency on how EPA arrived at the overall confidence level for each exposure hazard threshold.
Symbols are used to provide a visual overview of the confidence in the body of evidence, while de-
emphasizing an individual ranking that may give the impression that ranks are cumulative (e.g., ranks of
different categories may have different weights).

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TableApx C-l. Considerations that Inform Evaluations of the Strength of the Evidence within an Evidence Stream Apical
Endpoints, Mechanistic, or Field Studies)		

Consideration

Increased Evidence Strength (of the Apical
Endpoints, Mechanistic, or Field Studies
Evidence)

Decreased Evidence Strength (of the Apical Endpoints, Mechanistic, or
Field Studies Evidence)

The evidence considerations and criteria laid out here guide the application of strength-of-evidence judgments for an outcome or environmental hazard effect
within a given evidence stream. Evidence integration or synthesis results that do not warrant an increase or decrease in evidence strength for a given
consideration are considered "neutral" and are not described in this table (and, in general, are captured in the assessment-specific evidence profile tables).

Quality of the database'1
(risk of bias)

•	A large evidence base of high- or medium-quality
studies increases strength.

•	Strength increases if relevant species are
represented in a database.

•	An evidence base of mostly /ow-quality studies decreases strength.

•	Strength also decreases if the database has data gaps for relevant species,
i.e., a trophic level that is not represented.

•	Decisions to increase strength for other considerations in this table should
generally not be made if there are serious concerns for risk of bias; in other
words, all the other considerations in this table are dependent upon the
quality of the database.

Consistency

Similarity of findings for a given outcome (e.g., of a
similar magnitude, direction) across independent
studies or experiments increases strength,
particularly when consistency is observed across
species, life stage, sex, wildlife populations, and
across or within aquatic and terrestrial exposure
pathways.

•	Unexplained inconsistency (i.e., conflicting evidence; see U.S. EPA (2005)
decreases strength.)

•	Strength should not be decreased if discrepant findings can be reasonably
explained by study confidence conclusions; variation in population or
species, sex, or life stage; frequency of exposure (e.g., intermittent or
continuous); exposure levels (low or high); or exposure duration.

Strength (effect magnitude)
and precision

•	Evidence of a large magnitude effect (considered
either within or across studies) can increase strength.

•	Effects of a concerning rarity or severity can also
increase strength, even if they are of a small
magnitude.

•	Precise results from individual studies or across the
set of studies increases strength, noting that
biological significance is prioritized over statistical
significance.

•	Use of probabilistic model (e.g., Web-ICE, SSD)
may increase strength.

Strength may be decreased if effect sizes that are small in magnitude are
concluded not to be biologically significant, or if there are only a few
studies with imprecise results.

Biological gradient/dose-
response

•	Evidence of dose-response increases strength.

•	Dose-response may be demonstrated across studies
or within studies and it can be dose- or duration-
dependent.

• A lack of dose-response when expected based on biological
understanding and having a wide range of doses/exposures evaluated in the
evidence base can decrease strength.


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Consideration

Increased Evidence Strength (of the Apical
Endpoints, Mechanistic, or Field Studies
Evidence)

Decreased Evidence Strength (of the Apical Endpoints, Mechanistic, or
Field Studies Evidence)



•	Dose response may not be a monotonic dose-
response (monotonicity should not necessarily be
expected, e.g., different outcomes may be expected
at low vs. high doses due to activation of different
mechanistic pathways or induction of systemic
toxicity at very high doses).

•	Decreases in a response after cessation of exposure
(e.g., return to baseline fecundity) also may increase
strength by increasing certainty in a relationship
between exposure and outcome (this particularly
applicable to field studies).

•	In experimental studies, strength may be decreased when effects resolve
under certain experimental conditions (e.g., rapid reversibility after
removal of exposure).

•	However, many reversible effects are of high concern. Deciding between
these situations is informed by factors such as the toxicokinetics of the
chemical and the conditions of exposure, see (U.S. EPA, 1998). cndooint
severity, judgments regarding the potential for delayed or secondary
effects, as well as the exposure context focus of the assessment (e.g.,
addressing intermittent or short-term exposures).

•	In rare cases, and typically only in toxicology studies, the magnitude of
effects at a given exposure level might decrease with longer exposures
(e.g., due to tolerance or acclimation).

•	Like the discussion of reversibility above, a decision about whether this
decreases evidence strength depends on the exposure context focus of the
assessment and other factors.

•	If the data are not adequate to evaluate a dose-response pattern, then
strength is neither increased nor decreased.

Biological relevance

Effects observed in different populations or
representative species suggesting that the effect is
likely relevant to the population or representative
species of interest (e.g., correspondence among the
taxa, life stages, and processes measured or observed
and the assessment endpoint).

An effect observed only in a specific population or species without a clear
analogy to the population or representative species of interest decreases
strength.

Physical/chemical relevance

Correspondence between the substance tested and
the substance constituting the stressor of concern.

The substance tested is an analog of the chemical of interest or a mixture of
chemicals which include other chemicals besides the chemical of interest.

Environmental relevance

Correspondence between test conditions and
conditions in the region of concern.

The test is conducted using conditions that would not occur in the
environment.

" Database refers to the entire data set of studies integrated in the environmental hazard assessment and used to inform the strength of the evidence. In this context,
database does not refer to a computer database that stores aggregations of data records such as the ECOTOX Knowledgebase.

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