United States Office of Water EPA 440/5-80-067
Environmental Protection Regulations and Standards October 198C
Agency Criteria and Standards Division ,
Washington DC 20460 £,- I
4>EPA Ambient
Water Quality
Criteria for
Phthalate Esters
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AMBIENT WATER QUALITY CRITERIA FOR
PHTHALATE ESTERS
Prepared By
U.S. ENVIRONMENTAL PROTECTION AGENCY
Office of Water Regulations and Standards
Criteria and Standards Division
Washington, D.C.
Office of Research and Development
Environmental Criteria and Assessment Office
Cincinnati, Ohio
Carcinogen Assessment Group
Washington, D.C.
Environmental Research Laboratories
Corvalis, Oregon
Duluth, Minnesota
Gulf Breeze, Florida
Narragansett, Rhode Island
US ENVIRONMENTAL PROTECTION AGENCY
REGION 5 LIBRARY (PL-12J)
i 77 WEST JACKSON BLVD 12TH FLOOR
CHICAGO IL 60604-3590
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DISCLAIMER
This report has been reviewed by the Environmental Criteria and
Assessment Office, U.S. Environmental Protection Agency, and approved
for publication. Mention of trade names or commercial products does not
constitute endorsement or recommendation for use.
AVAILABILITY NOTICE
This document is available to the public through the National
Technical Information Service, (NTIS), Springfield, Virginia 22161.
11
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FOREWORD
Section 304 (a)(l) of the Clean Water Act of 1977 (P.L. 95-217),
requires the Administrator of the Environmental Protection Agency to
publish criteria for water quality accurately reflecting the latest
scientific knowledge on the kind and extent of all identifiable effects
on health and welfare which may be expected from the presence of
pollutants in any body of water, including ground water. Proposed water
quality criteria for the 65 toxic pollutants listed under section 307
(a)(l) of the Clean Water Act were developed and a notice of their
availability was published for public comment on March 15, 1979 (44 FR
15926), July 25, 1979 (44 FR 43660), and October 1, 1979 (44 FR 56628).
This document is a revision of those proposed criteria based upon a
consideration of comments received from other Federal Agencies, State
agencies, special interest groups, and individual scientists. The
criteria contained in this document replace any previously published EPA
criteria for the 65 pollutants. This criterion document is also
published in satisifaction of paragraph 11 of the Settlement Agreement
in Natural Resources Defense Council, et. al. vs. Train, 8 ERC 2120
(D.D.C. 1976), modified, 12 ERC 1833 (D.D.C. 1979).
The term "water quality criteria" is used in two sections of the
Clean Water Act, section 304 (a)(l) and section 303 (c)(2). The term has
a different program impact in each section. In section 304, the term
represents a non-regulatory, scientific assessment of ecological ef-
fects. The criteria presented in this publication are such scientific
assessments. Such water quality criteria associated with specific
stream uses when adopted as State water quality standards under section
303 become enforceable maximum acceptable levels of a pollutant in
ambient waters. The water quality criteria adopted in the State water
quality standards could have the same numerical limits as the criteria
developed under section 304. However, in many situations States may want
to adjust water quality criteria developed under section 304 to reflect
local environmental conditions and human exposure patterns before
incorporation into water quality standards. It is not until their
adoption as part of the State water quality standards that the criteria
become regulatory.
Guidelines to assist the States in the modification of criteria
presented in this document, in the development of water quality
standards, and in other water-related programs of this Agency, are being
developed by EPA.
STEVEN SCHATZOW
Deputy Assistant Administrator
Office of Water Regulations and Standards
111
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ACKNOWLEDGEMENTS
Aquatic Life ioxicology:
William A. Brungs, ERL-Narragansett
U.S. Environmental Protection Agency
David J. Hansen, ERL-Gulf Breeze
U.S. Environmental Protection Agency
Mammalian Toxicology and Human Health Effects:
John Autian (author)
University of Tennessee
Steven D. Lutkenhoff (doc. mgr.)
ECAO-Cin
U.S. Environmental Protection Agency
Bonnie Smith (doc. mgr.) ECAO-Cin
U.S. Environmental Protection Agency
Sherwin V. Kevy
Harvard Medical School
Krishna ?. Misra
U.S. Food and Drug Administration
Alan B. Rubin
U.S. Environmental Protection Agency
Herbert Schumacher
National Center for Toxicological
Research
Patrick Durkin
Syracuse Research Corporation
Karl Gabriel
Medical College of Pennsylvania
May Jacobson
Harvard Medical School
Van Kozak
U.S. Environmental Protection Agency
Bart Puma
U.S. Food and Drug Administration
Robert J. Rubin
Johns Hopkins University
James Withey
Health and Welfare, Canada
Technical Support Services Staff: D.J. Reisman, M.A. Garlough, B.L. Zwayer,
P.A. Daunt, K.S. Edwards, T.A. Scandura, A.T. Pressley, C.A. Cooper,
M.M. Denessen.
Clerical Staff: C.A. Haynes, S.J. Faehr, L.A. Wade, D. Jones, B.J. Bordicks
B.J. Quesnell, T. Highland, R. Rubinstein.
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TABLE OF CONTENTS
Page
Criteria Summary
Introduction A-l
Aquatic Life Toxicology B-l
Introduction B-l
Effects B-l
Acute Toxicity B-l
Chronic Toxicity B-3
Plant Effects B-3
Residues B-4
Miscellaneous B-5
Summary B-6
Criteria B-7
References B-20
Mammalian Toxicology and Human Health Effects C-l
Introduction C-l
Exposure C-2
Ingestion from Water C-2
Ingestion from Food C-2
Inhalation C-7
Dermal C-8
Pharmacokinetics C-12
Absorption C-12
Distribution C-13
Metabolism C-16
Excretion C-16
Effects C-19
Acute, Subacute, and Chronic Toxicity C-19
Synergism and/or Antagonism C-38
Teratogenicity C-38
Mutagenicity C-41
Carcinogenicity C-43
Other Biological Effects C-43
Criterion Formulation C-53
Existing Guidelines and Standards C-53
Current Levels of Exposure C-53
Special Groups at Risk C-55
Basis and Derivation of Criterion C-56
References C-62
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CRITERIA DOCUMENT
PHTHALATE ESTERS
CRITERIA
Aquatic Life
The available data for phthalate esters indicate that acute and chronic
toxicity to freshwater aauatic life occur at concentrations as low as 940
and 3 pg/1, respectively, and would occur at lower concentrations among spe-
cies that are more sensitive than those tested.
The available data for phthalate esters indicate that acute toxicity to
saltwater aauatic life occurs at concentrations as low as 2,944 yg/1 and
would occur at lower concentrations among species that are more sensitive
than those tested, No data are available concerning the chronic toxicity of
phthalate esters to sensitive saltwater aauatic life but toxicity to one
species of algae occurs at concentrations as low as 3.4 ug/1.
Human Health
For the protection of human health from the toxic properties of dimethyl
phthalate ingested through water and contaminated aauatic organisms, the
ambient water criterion is determined to be 313 mg/1.
For the protection of human health from the toxic properties of dimethyl
phthalate ingested through contaminated aauatic organisms alone, the ambient
water criterion is determined to be 2.9 g/1.
For the protection of human health from the toxic properties of diethyl
phthalate ingested through water and contaminated aauatic organisms, the
ambient water criterion is determined to be 350 mq/1.
For the protection of human health from the toxic properties of diethyl
phthalate ingested through contaminated aauatic organisms alone, the ambient
water criterion is determined to be 1.8 g/1.
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For the protection of human health from the toxic properties of dibutyl
phthalate ingested through water and contaminated aquatic organisms, the
ambient water criterion is determined to be 34 mg/1.
For the protection of human health from the toxic properties of dibutyl
phthalate ingested through contaminated aquatic organisms alone, the ambient
water criterion is determined to be 154 mg/1.
For .the protection of human health from the toxic properties of di-2-
ethylhexyl phthalate ingested through water and contaminated aquatic orga-
nisms, the ambient water criterion is determined to be 15 mg/1.
For the protection of human health from the toxic properties of di-2-
ethylhexyl phthalate ingested through contaminated aquatic organisms alone,
the ambient water criterion is determined to be 50 mg/1.
VII
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INTRODUCTION
Phthalic acid esters (PAEs), or "phthalate esters," represent a large
family of chemicals widely used as plasticizers, primarily in the production
of polyvinyl chloride (PVC) resins (U.S. Int. Trade Comm., 1978). Table 1
lists the major esters with their production figures. Phthalates are esters
of the ortho form of benzenedicarboxylic acid, also referred to as ortho-
phthalic acid. Two other isomeric forms of phthalic acid esters are also
produced. These include the meta form (or isothalate esters) and the para
form (or terephthalate esters). Both of these isomers have a number of
important commercial applications such as starting materials for plastics
and textiles. In this document, however, consideration will be given only
to the ortho-phthalate esters.
The annual production of phthalic acid esters in the United States in
1977 amounted to approximately 1.2 billion pounds. Since 1945, the cumula-
tive total production (up to 1972) of these esters reached a figure of 12.5
billion pounds (Peakall, 1975). On a worldwide scale, three to four billion
pounds are produced annually.
The most widely used phthalate plasticizer is di-2-ethylhexyl phthalate
(DEHP), which accounted for an estimated 32 percent of the total phthalate
esters produced in 1977 (U.S. Int. Trade Comm., 1978). In addition to DEHP,
other phthalates produced included other dioctyl phthalates, butylbenzyl
phthalate (BBP), diisodecyl phthalate, dibutyl phthalate (DBP), diethyl
phthalate (DEP), dimethyl phthalate (DMP), di-tridecyl phthalate, and
n-hexyl n-decyl phthalate (U.S. Int. Trade Comm., 1978).
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TABLE 1
Production of Individual Phthalic Acid
Esters in U.S. in 1977*
Ester Production in Pounds
(1,000 pounds)
Dibutyl
Diethyl
Diisodecyl
Dimethyl
Dioctyl
Di-2 -ethyl hexyl
Other dioctyl phthalates
Di-tridecyl
n-Hexyl n-decyl
All other phthalate esters
Total
16,592
17,471
160,567
9,887
388,543
11,664
23,278
15,182
559,229
1,202,413
*Source: United States International Trade Commission,
1978
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PVC resins are used in such diverse industries as construction (high
temperature electrical wire, cable insulation, and flooring), home furnish-
ings (furniture upholstery, wall coverings), transportation (upholstery and
seat covers), apparel (footwear), and food and medical packaging materials.
Phthalates also have non-plasticizer uses in pesticide carriers, cosmetics,
fragrances, munitions, industrial oils, and insect repellants (U.S. Int.
Trade Comm., 1978). Table 2 illustrates the variety of uses for esters with
an estimate of the amount of the esters used in the specific categories.
PAE plasticizers can be present in concentrations up to 60 percent of
the total weight of the plastic. The plasticizers are loosely linked to the
plastic polymers and are easily extracted (Mathur, 1974).
For the most part, the esters are colorless liquids, have low volatil-
ity, and are poorly soluble in water but soluble in organic solvents and
oils. Table 3 lists several of the physical properties of these esters.
The phthalate esters can be prepared by reaction of phthalic acid with a
specific alcohol to form the desired esters. In industry, however, the
esters are manufactured from phthalic anhydride rather than from the acid.
For the most part, manufactured esters will not be completely pure, having
various isomers and contaminants present. These esters, however, can be
prepared with a purity of greater than 99 percent even though most of these
esters are not sold with this high degree of purity.
Evidence also is available suggesting that certain plants and animal
tissue may synthesize phthalic acid esters (Peakall, 1975). However, to
what extent this occurs in nature is not known.
The ease of extraction of phthalate esters and their widespread use
either alone or in PVC account for their ubiquity. PAEs have been detected
in soil (Ogner and Schnitzer, 1970), water (Ewing and Chian, 1977; Corcoran,
A-3
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TABLE 2
Uses of Phthalate Esters in the United States*
A. As Plasticizers
Building and Construction
Wire and cable 185
Flooring 150
Swimming pool liners 20
Miscellaneous 32
Subtotal 387
Home Furnishings
Furniture upholstery 90
Wall coverings 38
Houseware 30
Miscellaneous 45
Subtotal 20?
Cars (upholstery, tops, etc.) 114
Wearing apparel 72
Food wrapping and closures 25
Medical tubing and intravenous bags 21
Total as Plasticizers 922
B. As Nonplasticizers
Pesticide Carriers
Oils __
Insect repellent
Total as Nonplasticizers .. 50
Grand Total 972
*Source: Graham, 1973
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TABLE 3
Physical and Chemical Properties of Phthalate Esters
Compound Molecular
Weight
Dimethyl
phthalate
Oi ethyl
phthalate
Diallyl
phthalate
Diisobutyl
phthalate
01 butyl
phthalate
Dimethoxyethyl
phthalate
Dicyclohexyl
phthalate
Butyl octyl
phthalate
Dihexyl
phthalate
Butylphthalyl
butyl glycolate
Dibutoxyethyl
ethyl phthalate
Oi-2-ethylhexyl
phthalate
Diisooctyl
phthalate
Oi-n-octyl
phthalate
Dinonyl
phthalate
194.18
222.23
246.27
278.3
278.34
282.0
330.0
334.0
334.0
336.37
366.0
391.0
391.0
391.0
419.0
Specific
Gravity
1.189
1.123
1.120
1.040
1.0465
1.171
1.20
—
0.990
1.097
1.063
0.985
0.981
0.978
0.965
BP, Solubility in
°C H20, g/100 ml
282
296.1
290
327
340
190-210
220-228
340
—
219*
210
386.9*
239*
220*
413
0.5
Insoluble
0.01
Insoluble
0.45 (25°C)
0.85
Insoluble
—
Insoluble
0.012%
0.03
Insoluble
Insoluble
Insoluble
Insoluble
*Measured at 5 mm Hg
A-5
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1973; Hites and Bieman, 1972), fish (Mayer, 1976; Stalling, 1973), air
(Mathur, 1974) and animal and human tissues (Nazir, et al. 1971; Rubin and
Shiffer, 1976; Jaeger and Rubin, 1970). Their detection in certain vegeta-
tion, animals and minerals (Mathur, 1974; Graham, 1973), and in areas remote
from industrial sites (Carpenter and Smith, 1972) have raised questions
about possible natural origins of PAEs. PAEs found in greatest frequencies
in an EPA monitoring survey of U.S. surface waters (Ewing and Chian, 1977)
were DEHP (132/204) and DEP (84/204). Other esters detected in the EPA sur-
vey were diethyl phthalate, disobutyl phthalate, and diocyl phthalate.
A-6
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REFERENCES
Carpenter, E. and K. Smith. 1972. Plastics on the Sargasso sea surface.
Science. 175: 1240.
Cocroran, E. 1973. Gas chromatographic detection of phthalate acid esters.
Environ. Health Perspect. 3: 13.
Ewing, B. and E. Chian. 1977. Monitoring to detect previously unrecognized
pollutants in surface waters. EPA 560/7-77/15a. Off. Tox. Subst., U.S.
Environ. Prot. Agency, Washington, D.C.
Graham, P. 1973. Phthalate ester plasticizers - why and how they are used.
Environ. Health Perspect. 3: 3.
Hites, R. and K. Bieman. 1972. Water pollution - organic compounds in the
Charles River, Boston. Science. 178: 158.
Jaeger, R. and R. Rubin. 1970. Plasticizers from plastic derivatives.
Exhaustion, metabolism, and accumulation by biological systems. Science.
170: 460.
Mathur, S. 1974. Phthalate esters in the environment: Pollutants or natu-
ral products? Jour. Environ. Quality. 3: 189.
Mayer, F.L. 1976. Residue dynamics of di-2-ethylhexylphthalate in fathead
minnows, Pimephales promelas. Jour. Fish. Res. Board Can. 33: 2610.
A-7
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Nazir, D., et al. 1971. Isolation, identification, and specific localiza-
tion of di-2-ethylhexyl phthalate- ,in bovine heart muscle mitochondria. Bio-
chem. 10: 4425.
Ogner, G. and M. Schnitzer. 1970. Humic substances: Fulvic acid - dialkyl
phthalate complexes and their role in pollution. Science. 170: 317.
Peakall, D. 1975. Phthalate esters: Occurrence and biological effects.
Residue Rev. 54: 1.
Rubin, R. and C. Schiffer. 1976. Fate in humans of the plasticizer, di-2-
ethylhexyl phthalate, arising from platelets stored in vinyl plastic bags.
Transfusion. 16: 330.
Stalling, D., et al. 1973. Phthalate ester residues - their metabolism and
analysis in fish. Environ. Health Perspect. 3: 159.
U.S. International Trade Commission. 1978. Synthetic organic chemicals,
U.S. production and sales. Washington, D.C.
A-8
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Aquatic Life Toxicology*
INTRODUCTION
Phthalate esters are a large group of chemical agents (esters of ortho
benzene dicarboxylic acid) used primarily as plasticizers.
A limited number of applicable reports were found having effects data on
individual phthalate esters to freshwater aquatic life. More information is
available for butylbenzyl and di-2-ethylhexyl phthalate than for the other
esters.
Toxicity test data for saltwater organisms are available for six phtha-
late esters. Tests have provided some acute and plant effects of butylbenzyl
phthalate, diethyl phthalate, and dimethyl phthalate. Limited information is
also available on di-n-propyl, di-n-butyl, and di-2-ethylhexyl phthalates.
These data indicate great differences in toxicity among esters.
EFFECTS
Acute Toxicity
All freshwater acute values were determined with static procedures and
the test concentrations were unmeasured. Data for five phthalate esters can
be found in Table 1. Values for four of the esters were from tests with both
fish and invertebrate species.
Tests with butylbenzyl, diethyl, and dimethyl phthalate were conducted
with bluegill, fathead minnow, and Daphnia magna (U.S. EPA, 1978; Sledhill, et
al. 1980). The acute values ranged from 1,700 to 98,200 ug/1.
Gledhill, et al. (1980) reported butylbenzyl phthalate LCgQ values for
three fish and one invertebrate species. The values ranged from 1,700 to
*The reader is referred to the Guidelines for Deriving Water Quality Criteria
for the Protection of Aquatic Life and Its Uses in order to better understand
the following discussion and recommendation. The following tables contain the
appropriate data that were found in the literature, and at the bottom of the
appropriate table are calculations for deriving various measures of toxicity
as described in the Guidelines.
8-1
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5,300 ug/1 (Table 1). The two fathead minnow acute values represent two water
hardness levels. Their IC™ values for Daphnia magna and bluegills were
about 25 times less than reported by the U.S. EPA (1978).
Di-n-butyl phthalate tests were conducted with four fish and two inverte-
brate species. The IC™ values varied from 730 to 6,470 pg/1 or a differ-
ence of about nine times. Bluegills were the most sensitive fish and the scud
the most sensitive invertebrate species tested with this ester. An additional
acute datum for a crayfish species and this ester is included in Table 6, but
the LCrQ value exceeded the highest test concentration (10,000 ug/1).
Only one acute value was obtained with di-2-ethylhexyl phthalate and was
derived from a test with Daphnia magna. Additional acute data for this ester
are shown in Table 6, and the LC5Q values for the midge, scud, and bluegill
exceeded the highest concentrations tested. The LCrQ range for Daphnia
magna (Monsanto, 1978) represents the 50 percent mortalities obtained in two
of the six concentrations.
Acute effects of only three phthalate esters (butylbenzyl phthalate,
diethyl phthalate, and dimethyl phthalate) on two saltwater species, mysid
shrimp and sheepshead minnow, have been reported (Table 1). All of the eight
data were based on static test procedures with unmeasured concentrations. For
the effects of butylbenzyl phthalate, there was a great difference between the
two values for the mysid shrimp (900 and 9,630 ug/1) and also between those
for the sheepshead minnow (3,000 and 445,000 ug/1). The tests were conducted
by the same laboratory, but the lower values were obtained in tests using a
solvent (Gledhill, et al. 1980) and the higher values represent tests not us-
ing a solvent (U.S. EPA, 1978). Undoubtedly, much of the chemical was not
available to the test animals when a solvent was not used. Less than full
solubility of the chemical may also have occurred for the data on diethyl
phthalate (7,590 ug/1 for mysid shrimp; 29,600 yg/l for sheepshead minnow) and
B-2
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dimethyl phthalate (73,700 ug/1 for mysid shrimp; 58,000 ug/1 for sheepshead
minnow) generated without use of solvents (U.S. EPA, 1978), although there are
no comparable data obtained using solvents.
Chronic Toxicity
Freshwater data were found for two phthalate esters and the results are
presented in Table 2. An early life stage test with a fish and a life-cycle
Daphnia magna test were conducted for each ester.
The butylbenzyl phthalate chronic values reported for the fathead minnow
and Daphnia magna were 220 and 440 ug/1, respectively. The corresponding
acute-chronic ratios were determined to be 17 and 42. The chronic values and
acute-chronic ratios for this ester were within a factor of about 2 for the
fish and invertebrate species.
A di-2-ethylhexyl phthalate test was conducted with rainbow trout. The
chronic value was 8.4 ug/l. No acute-chronic ratio could be calculated be-
cause of the absence of a 96-hour |_C50 value. Mayer and Sanders (1973) con-
ducted a chronic test with di-2-ethylhexyl phthalate and Daphnia magna. Sign-
ificant reproductive impairment was found at 3 ug/1. Since this value was at
the lowest test concentration, the adverse effects on reproduction were less
than 3 ug/1. This concentration represents the lowest toxicity value reported
for the phthalate esters.
Species mean acute values and acute-chronic ratios are summarized in
Table 3.
No saltwater fish or invertebrate species have been tested in a chronic
toxicity study.
Plant Effects
The adverse effects of three phthalate esters on freshwater algal species
are summarized in Table 4. Similar EC^Q values with Selenastrum capricornu-
tum were found for cell numbers and chlorophyll a_ for each ester tested by the
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U.S. EPA (1978). By comparison, the butylbenzyl phthalate EC value found
by Gledhill, et al. (1980) with this alga was about 3 times higher, and the
alga, Microcystis aeruginosa, was shown to be resistant to this ester. The
lowest EC5Q values for diethyl and dimethyl phthalate were 85,600 and 39,800
yg/1, respectively. A much lower EC5Q value of 110 wg/l was obtained with
butylbenzyl phthalate, and represents a lower value than found for fish and
invertebrate species (Table 1).
Data on the toxicity of five phthalate esters to one or two species of
saltwater algae are listed in Table 4. Butylbenzyl phthalate and dimethyl
phthalate were more toxic to a saltwater alga, Skeletonema costatum, than to
the tested fish and invertebrate species.
The various phthalates showed a wide range of toxicity to the same spe-
cies of alga. Thus, butylbenzyl phthalate was very toxic to Skeletonema cost-
atum with a chlorophyll £ EC5Q value of 170 ug/l; however, the chlorophyll a
EC5Q of diethyl phthalate for the same species was 65,500 yg/1. In addi-
tion, the lowest EC5Q of di-n-butyl phthalate for Gymnodinium breve was 3.4
ug/1 and the lowest EC5Q of dimethyl phthalate for the same species was
54,000 ug/1. Some of these wide ranges in toxicity could be due to EC
values reported that may surpass the solubility limits of the compounds tested
or to relatively large differences reported for replicate tests, particularly
those of Wilson, et al. (1978).
Residues
Freshwater bioconcentration factors for five phthalate esters are report-
ed in Table 5. Mayer (1976) measured both the actual concentrations and
14
C-labeled di-2-ethylhexyl phthalate in a test system and found the differ-
ence was less than two times after equilibrium in fathead minnows. The bio-
concentration factors for di-2-ethylhexyl phthalate with fish and invertebrate
species ranged from 54 to 2,680. Tests with di-n-butyl phthalate performed
B-4
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with two invertebrate species gave equilibrium bioconcentration factors of 400
and 1,400. Bioconcentration factors for C-labeled butylbenzyl, diethyl,
and dimethyl phthalate with bluegills were 663, 117, and 57, respectively,
after a 21-day exposure (U.S. EPA, 1978). The half-life of these three phtha-
late esters was between 1 and 2 days. Bioaccumulation data with di-n-octyl
phthalates by Sanbom, et al. (1975) in a static model ecosystem are given in
Table 6. Their water concentrations rapidly decreased with time and do not
permit comparisons with values in Table 5.
Since no maximum permissible tissue levels exist for phthalate esters, no
Residue Limited Toxicant Concentration could be calculated for any phthalate
ester.
No data are available for bioconcentration of phthalate esters by any
saltwater species.
Miscellaneous
Additional freshwater toxicity data for phthalate esters are given in
Table 6. Many of these data have already been discussed and were not lower
than the acute or chronic values (Tables 1 and 2). Mayer, et al. (1977) ex-
posed rainbow trout embryos to di-2-ethylhexyl phthalate for 90 days and found
concentrations of 14 to 54 yg/1 significantly increased total protein catabol -
ism 24 days after hatching. This concentration range is similar to the lowest
adverse test concentration found with this ester in the embryo-larval test
(Table 2). Birge, et al. (1978) performed tests with several fish species
using di-isononyl and di-n-octyl phthalate. The tests were started with 7-
hour-old fertilized embryos and continued through four days post-hatch; be-
cause of the test duration and endpoints measured, the data for these two es-
ters were listed in Table 6. Also listed in this table is a diet study with
the guppy using di-2-ethylhexyl phthalate which resulted in an increase in
aborted young.
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Saltwater data for effects (Table 6) not listed in the other tables sug-
gest no more sensitive effects than those already presented. The only toxi-
city data available for di-2-ethylbutyl phthalate indicated that 1,000 Mg/l
had no significant effect on the entire larval development of the grass
shrimp, and that only a high concentration (EC5Q = 3.1%) affected growth
rate of the alga, Gymnodinium breve (Wilson, et al. 1978).
Summary
Acute freshwater test results were available for five phthalate esters,
and these were conducted with a relatively small diverse group of freshwater
fish and invertebrate species. The acute values, with one exception, all ex-
ceeded 1,000 ug/1. Sensitivity differences were generally similar for the
tested freshwater species. No final acute values are calculable for any ester
since the minimum data base requirements were not met.
Chronic freshwater test results were available for two phthalate esters.
The chronic values for butylbenzyl phthalate were 220 and 440 ug/1 with the
calculated acute-chronic ratios being 17 and 42. The chronic values for di-2-
ethylhexyl phthalate were 3 and 8.4 ug/l and no acute-chronic ratios were
calculable. No final chronic values could be determined.
Plant test results were available for three phthalate esters. The plant
values for diethyl and dimethyl phthalate were similar to the acute results
for these phthalates and invertebrate species. A wide variation was found in
the EC5Q values for butylbenzyl phthalate, which values ranged from 110 to
1,000,000 ug/1.
Residue test results were available for five phthalate esters. A wide
variation was found for bioconcentration values for both the invertebrate (14-
2,680) and fish (42-886) species. More residue data were available for di-2-
ethylhexyl phthalate than for the other esters.
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Additional freshwater toxicity results were available with four phthalate
esters. None of these data showed toxicity values lower than those already
discussed.
Acute saltwater test results were available for three phthalate esters
with one invertebrate and one fish species. The lowest concentrations at
which acute effects were observed were 900 ug/1 for butylbenzyl phthalate and
7,590 ug/1 for diethyl phthalate, both for mysid shrimp, and 58,000 ng/1 for
dimethyl phthalate with the sheepshead minnow. There were no saltwater chron-
ic or residue test results for any phthalate ester. Effects of phthalate
esters on saltwater algal species were reported at concentrations as low as
3.4 ug/l.
CRITERIA
The available data for phthalate esters indicate that acute and chronic
toxicity to freshwater aquatic life occur at concentrations as low as 940 and
3 ug/l, respectively, and would occur at lower concentrations among species
that are more sensitive than those tested.
The available data'for phthalate esters indicate that acute toxicity to
saltwater aquatic life occurs at concentrations as low as 2944 ug/l and would
occur at lower concentrations among species that are more sensitive than those
tested. No data are available concerning the chronic toxicity of phthalate
esters to sensitive saltwater aquatic life but toxicity to one species of
algae occurs at concentrations as low as 3.4 u
B-7
-------
Table 1. Acute values for phthaiate esters
Spec 1 es
Cladoceran,
Daphnla magna
C ladoceran,
Daphnla maqna
Cladoceran,
Daphnla maqna
Cladoceran,
Daphnla maqna
Cladoceran,
Daphnla maqna
Scud,
Gammarus pseudol Imnaeus
UU - — .
m Midge,
Chironomus plumosus
Rainbow trout.
Sal mo galrdnerl
Rainbow trout,
Sal mo galrdnerl
Fathead minnow,
Pimephales promelas
Fathead minnow,
Pimephales promelas
Fathead minnow,
Pimephales promelas
Bluegil 1,
Lepomis macrochirus
Bluegi 1 1,
Lepomis macrochirus
Method*
s.
s.
s.
s.
s.
s,
s,
s,
s,
s.
s,
s,
s,
s,
u
u
u
u
u
u
u
u
u
u
u
u
u
u
Chemical
FRESHWATER
Buty 1 benzyl
phthalate
Buty 1 benzyl
phthalate
01 ethyl
phthalate
Dimethyl
phthalate
di-2-ethy Ihexyl
phthalate
di-n-buty 1
phthalate
di-n-buty 1
phthalate
Buty 1 benzyl
phtha 1 ate
di-n-buty 1
phthalate
Buty 1 benzyl
phthalate
Buty 1 benzy 1
phthalate
di-n-butyl
phthalate
Buty 1 benzy 1
phthalate
Butyl benzyl
phthalate
LC50/EC50
(ug/D
Species Mean
Acute Value
(uq/l)
Reference
SPECIES
92.
3,
52,
33,
U.
2,
4,
3,
6.
5,
2,
1,
43,
1,
300
700
100
000
100
100
000
300
470
300
100
300
300
700
-
18,500
52,100
33,000
11,100
2,100
4,000
3,300
6,470
3,300
1,300
8,600
U.S. EPA
Gledhl 1 1
, 1978
, et al.
1980
U.S. EPA, 1978
U.S. EPA
, 1978
U.S. EPA, 1978
Mayer & Sanders,
Streufert, 1977
Gledhlll, et al.
Mayer & Sanders,
Gledhil 1, et al.
Gledhil 1
Mayer &
U.S. EPA
Gledhlll
, et al.
Sanders,
, 1978
, et al.
1973
1980
1973
1980
1980
1973
1980
-------
Table 1. (Continued)
to
i
Species Method*
Bluegi 1 1, S, U
Lepomls macrochirus
Blueglll, S, U
Lepomis macrochirus
Bluegi II, S, U
Lepomls macrochirus
Bluegi II, S, U
Lepomls macrochirus
Channel catfish, S, U
Ictalurus punctatus
Mysid shrimp, S, U
Mysidopsis bahia
Mysid shrimp, S, U
Mysidopsis bahia
Mysid shrimp, S, U
Mysidopsis bahia
Mysid shrimp, S, U
Mysidopsis bahia
Sheepshead minnow, S, U
Cyprinodon varlegatus
Sheepshead minnow, S, U
Cyprinodon varlegatus
Sheepshead minnow, S, U
Cyprinodon variegatus
Sheepshead minnow, S, U
Cyprinodon variegatus
Chemical
Diethy 1
phthalate
Dimethyl
phthalate
dl-n-buty 1
phthalate
di-n-buty 1
phthalate
di-n-buty 1
phthalate
SALTWATER
Butyl benzy 1
phthalate
Buty (benzyl
phthalate
Di ethyl
phthalate
Dimethyl
phthalate
Buty 1 benzyl
phthalate
Butyl benzyl
phthalate
Di ethyl
phthalate
Dimethyl
phthalate
LC50/EC50
-------
Table 2. Chronic values for phthalate esters
DO
I
Species
Cladoceran,
Daphnla magna
Cladoceran,
Daphnla magna
Rainbow trout,
Salmo gairdneri
Method*
LC
LC
ELS
Fathead minnow, ELS
Plmephales promelas
* ELS = early 1
Ife stage, LC = partial
Species
Cladoceran,
Daphnla magna
Cladoceran,
Daphnla magna
Rainbow trout,
Salmo galrdneri
Fathead minnow,
Pimephales promelas
Chemical
FRESHWATER
Butyl benzyl
phthalate
dl-2-ethy Ihexyl
phthalate
dl-2-ethy Ihexyl
phthalate
Buty (benzyl
phthalate
Limits
(ug/l)
SPECIES
260-760
<3
5-14
140-360
Species Mean
Chronic Value
(ug/ 1 ) Reference
440 Gledhil 1, et al. 1980
<3 Mayer & Sanders, 1973
8.4 Mehrle & Mayer, 1976
220 U.S. EPA, 1978
life cycle or full life cycle
Acute-Chronic Ratios
Chemical
Buty (benzyl
phthalate
di-2-ethy Ihexyl
phthalate
dl-2-ethy Ihexyl
phthalate
Buty (benzyl
phthalate
Acute
Value
(ug/l)
18,500
11,100
3,300
Chronic
Value
(ug/l) Ratio
440 42
<3
8.4
220 15
-------
Table 3. Species mean acute values and acute-chronic ratios for phthalate esters
CO
I
Rank*
16
15
14
13
12
11
10
Species
Bluegi 1 1,
Lepomls macrochlrus
Cladoceran,
Daphnla magna
Bluegi II,
Lepomls macrochlrus
Cladoceran,
Daphnia magna
Cladoceran,
Daphnia magna
Cladoceran,
Daphnia magna
Bluegi 1 1,
Lepomls macrochlrus
Rainbow trout,
Sal mo gairdnerl
Midge,
Chironomus plumosus
Fathead minnow,
Pimephales promelas
Rainbow trout,
Sa Imo gairdner i
Channel catfish,
Ictalurus punctatus
Scud,
Gammarus pseudol Imnaeus
Fathead minnow,
Pimephales promelas
Chemical
FRESHWATER SPECIES
Di ethyl
phtha late
DI ethyl
phthalate
Dlmethy 1
phtha late
Dimethyl
phthalate
Butyl benzy 1
phtha late
dI-2-ethylhexyl
phtha late
Buty 1 benzy 1
phthalate
di -n-buty 1
phthalate
di -n-buty 1
phtha late
Butyl benzyl
phthalate
Buty 1 benzy 1
phthalate
di -n-buty 1
phtha late
d i-n-buty 1
phtha late
di-n-buty 1
phtha late
Species Mean
Acute Value
(ug/l)
96,200
52,100
49,500
33,000
18,500
1 1 , 100
8,600
6,470
4,000
3,300
3,300
2,910
2,100
1,300
Acute-Chronic
Ratio
42
15
-------
Table 3. (Continued)
CD
I
Rank*
2
1
6
5
4
3
2
1
Species
Bluegl 1 1,
Lap OBI Is macroch irus
Rainbow trout.
Sal mo galrdnerl
Mysid shrimp,
Mysidopsls bahja^
Sheepshead minnow,
Cyprinodon varleqatus
Sheepshead minnow,
Cyprinodon varlegatus
Sheepshead minnow,
Cyprinodon varleqatus
Mysid shrimp,
Mysidopsls bahia
Mysid shrimp,
Mysldopsis bah la
Chemical
di-n-buty 1
phthalate
d!-2-ethy Ihexyl
phthalate
SALTWATER SPECIES
Dimethyl
phthalate
Dimethyl
phthalate
Butyl benzyl
phthalate
Dl ethyl
phthalate
Di ethyl
phthalate
Butyl benzyl
phthalate
Species Mean
Acute Value
-------
Table 4. Plant values for phthalate esters
W
I
M
U>
Species
Chemical
Effect
Result
(ug/l) Reference
FRESHWATER SPECIES
Alga,
Selenastrum capricornutum
Alga,
Selenastrum capricornutum
Alga,
Selenastrum capricornutum
Alga,
Selenastrum capricornutum
Alga,
Selenastrum capricornutum
Alga,
Selenastrum capricornutum
Alga,
Selenastrum capricornutum
Alga,
Microcystis aeruginosa
Alga,
Navicula pelliculosa
Alga,
Skeletonema costatum
Alga,
Skeletonema costatum
Alga,
Skeletonema costatum
Alga,
Skeletonema costatum
Butyl benzyl
phthalate
Butyl benzyl
phthalate
Butyl benzy 1
phthalate
D i ethy 1
phthalate
Dlethy 1
phtha late
Dimethyl
phthalate
Dimethyl
phtha late
Buty 1 benzy 1
phthalate
Buty 1 benzyl
phtha late
Buty 1 benzy 1
phthalate
Buty 1 benzy 1
phthalate
Buty 1 benzy 1
phthalate
Dl ethyl
phthalate
96-hr EC50
chlorophy 1 1 a
96- hr EC50
ce 1 1 number
96-hr EC50
ce 1 1 number
96- hr EC50
ch lorophy 1 1 a
96- hr EC50
ce 1 1 number
96-hr EC50
ch lorophy 1 1 a
96-hr EC50
eel 1 number
96-hr EC50
eel 1 number
96- hr EC50
eel 1 number
SALTWATER SPECIES
96- hr EC 50
ch lorophy 1 1 a
96- hr EC 50
eel 1 number
96- hr EC 50
eel 1 number
96- hr EC 50
ch lorophy 1 1 a
1 10 U.S. EPA, 1978
130 U.S. EPA, 1978
400 Gledhi II, et al. 1980
90,300 U.S. EPA, 1978
85,600 U.S. EPA, 1978
42,700 U.S. EPA, 1978
39,800 U.S. EPA, 1978
1,000,000 Gledhi II, et al. 1980
600 Gledhi II, et al. 1980
170 U.S. EPA, 1978
190 U.S. EPA, 1978
600 Gledhill, et al. 1980
65,500 U.S. EPA, 1978
-------
Table 4. (Continued)
Species
Chemical
Effect
Result
(ug/I) Reference
Alga,
Skeletonema costatum
Alga,
Skeletonema costatum
Alga,
Skeletonema costatum
Alga,
Gymnodinium breve
Alga,
Gymnodinium breve
Alga,
Gymnodinium breve
CO Alga,
1 Gymnodinium breve
M
** Alga,
Gymnodinium breve
Alga,
Gymnodinium breve
Alga,
Gymnodinium breve
Alga,
Gymnodinium breve
Alga,
Gymnodinium breve
Alga,
Gymnodinium breve
Alga,
Gymnodinium breve
Di ethyl
phthalate
Dimethyl
phthalate
Dimethyl
phthalate
Di ethyl
phthalate
Di ethyl
phthalate
DI ethyl
phthalate
Dimethyl
phthalate
Dimethyl
phthalate
Dimethyl
phthalate
Dimethyl
phthalate
di-n-buty 1
phthalate
di-n-buty 1
phthalate
di-n-butyl
phthalate
di-n-butyl
phthalate
96- hr EC 50
eel 1 number
96-hr EC50
ch lorophy 1 1 a
96- hr EC50
eel 1 number
96- hr EC50
ch lorophy 1 1 a
96- hr EC50
ch lorophy 1 1 a
96-hr EC50
eel 1 number
96- hr EC50
ch lorophy 1 1 a
96-hr EC50
ch lorophy 1 1 a
96- hr EC 50
eel 1 number
96- hr EC50
eel 1 number
96- hr EC50
ch lorophy 1 1 a
96- hr EC 50
chlorophyll a
96-hr EC50
eel 1 number
96- hr EC50
eel 1 number
85,000 U.S. EPA, 1978
26,100 U.S. EPA, 1978
29,800 U.S. EPA, 1978
6,100 Wilson, et al. 1978
3,000 Wilson, et al. 1978
33,000 Wilson, et al. 1978
96,000 Wilson, et al. 1978
54,000 Wilson, et al. 1978
125,000 Wilson, et al. 1978
185,000 Wilson, et al. 1978
200 Wilson, et al. 1978
3.4 Wilson, et al. 1978
600 Wilson, et al. 1978
20 Wilson, et al. 1978
-------
Table 4. (Continued)
to
I
M
(Jl
Species
Chemical
Effect
Result
(ug/I) Reference
Alga,
Gymnodln ium breve
Alga,
Gymnodln Ium breve
Alga,
Gymnodln ium breve
Alga,
Gy mnod 1 n 1 urn bre ve
Alga,
Duna 1 1 e 1 1 a tert i o 1 ecta
di-n-propyl
phthalate
dl-n-propy 1
phthalate
d 1 -n-propy 1
phthalate
dl -n-propy 1
phthalate
Buty 1 benzy 1
phthalate
96- hr EC50
ch lorophy 1 1 a
96- hr EC50
ch lorophy 1 1 a
96- hr EC 50
eel 1 number
96-hr EC50
eel 1 number
96- hr EC50
eel 1 number
2,400
900
6,500
1.300
1,000
Wilson,
Wilson,
Wilson,
Wilson,
Gledhll
et al. 1978
et al. 1978
et al. 1978
et al. 1978
1, et al. 1980
-------
Table 5. Residues for phthalate esters
Tissue
Chemical
BIoconoentrat i on
Factor*
Duration
(days) Reference
FRESHWATER SPECIES
Cladoceran,
Daphnla magna
Scud,
Gammarus pseudol imnaeus
Scud,
Gammarus pseudol imnaeus
Sow bug,
Asel lus brevlcaudus
Rainbow trout,
Sal mo qalrdneri
CD Fathead minnow,
' Pimephales promelas
en
Fathead minnow,
Pimephales promeias
Bluegil 1,
Lepomis macrochirus
B 1 ueg i 1 1 ,
Lepomis macrochirus
Bluegil 1,
Lepomis macrochirus
Whole body
Whole body
Whole body
Whole body
Whole body
Whole body
Whole body
Whole body
Whole body
Whole body
di-n-butyl
phthalate
di-n-butyl
phthalate
di-2-ethylhexyl
phthalate
di-2-ethylhexyl
phthalate
di-2-ethylhexyl
phthalate
dl-2-ethylhexyl
phthalate
dl-2-ethylhexyl
phthalate
Butyl benzyl
phthalate
Dl ethyl
phthalate
Dimethyl
phthalate
400 14 Mayer & Sanders, 1973
1,400 14 Mayer & Sanders, 1973
54-2,680** 14-21 Sanders, et al. 1973
14-50** 21 Sanders, et al. 1973
42-113 36 Mehrle 4 Mayer, 1976
155-886 56 Mayer, 1976
91-569*** 56 Mayer, 1976
663 21 U.S. EPA, 1978
117 21 U.S. EPA, 1978
57 21 U.S. EPA, 1978
* Based on total '^C radioactivity accumulated.
** Conversion from dry to wet weight.
"""Based on measured concentrations of di-2-ethylhexyl phthalate.
-------
Table 6. Other data for phthaiate esters
Species
Alga,
Oedogonium cardiacum
Cl adoceran,
Daphnla magna
Cl adoceran,
Daphnla magna
Midge,
Chironomus plumosus
Scud,
Gammarus pseudol (mnaeus
03
1 Mosquito (larva),
|7J Culex pi pens
qu i nquef ascl at us
Snai 1,
Physa sp.
Crayfish,
Orconectes nais
Rainbow trout,
Sa Imo gairdner i
Ra i nbow trout
(early 1 ife stage),
Sa Imo qairdner i
Rainbow trout,
Sa I/no gairdfier i
Chemical
di-n-octy 1
phtha late
di-2-ethylhexyl
phtha late
di-n-octy 1
phtha late
d i-2-ethy 1 hexy 1
phtha late
d I-2-ethy Ihexy 1
phtha late
di-n-octy 1
phtha late
dl-n-octy 1
phtha late
di-n-buty 1
phtha late
di-2-ethylhexyl
phthalate
d i-n-octy 1
phthalate
di-n-octy 1
phthalate
Duration
FRESHWATER
33 days
48 hrs
33 days
48 hrs
96 hrs
33 days
33 days
96 hrs
24 days
26 days
26 days
Result
Effect (yg/l)
SPECIES
Model ecosystem*
28,50QX
bloconcentrat ion
LC50 1,000-
5,000
Model ecosystem*
2,600X
b 1 oconcentrat i on
LC50 > 18, 000
LC50 >32,000
Model ecosystem*
9.400X
bi oconcentrat ion
Model ecosystem*
I3,600X
bloconcentrat ion
LC50 > 10, 000
Significant 14-54
Increase in total
body protein
catabol ism
LC50 139,500
LC50 149,200
Reference
Sanborn, et al. 1975
Monsanto, 1978
Sanborn, et al. 1975
Streufert, 1977
Sanders, et al . 1973
Sanborn, et al. 1975
Sanborn, et al. 1975
Mayer & Sanders, 1973
Mayer, et al . 1977
Birge, et al. 1978
Birge, et al. 1978
-------
Table 6. (Continued)
Species
Guppy,
Poecilia retlculata
Bluegi 1 1,
Lepomls macrochirus
Redear sunf i sh
(early life stage),
Lepomls mlcrolopus
Redear sunfish,
(early 1 1 f e stage),
Lepomls mlcrolopus
Mosqultof ish,
Garnbusia affinls
1 Channel catfish
*~" (early life stage),
00 1 ctalurus punctatus
Channel catfish
(early life stage),
1 ctalurus punctatus
Largemouth bass
(early life stage),
Micropterus sal mo ides
Largemouth bass
(early 1 i fe stage),
Micropterus sal mo ides
Alga,
Gymnodinium breve
Chemical
di-2-ethy Ihexy 1
phthal ate
di-2-ethylhexyl
phthalate
dl-lsononyl
phthalate
dl-n-octy 1
phthalate
dl-n-octy 1
phthalate
dl-isonony 1
phthalate
dl-n-octy 1
phthalate
di-n-octy 1
phthalate
di-n-octy 1
phthalate
di-2-ethylhexyl
phtha late
Duration
90 days
96 hrs
7-8 days
7-8 days
33 days
7 days
7 days
7-8 days
7-8 days
SALTWATER
96 hrs
Result
Effect (ug/»)
Increase in fed 100
aborted young ug/g in
diet
LC50 >770,000
LC50 4,670
LC50 6,180
Model ecosystem*
9.400X
bi oconcentrat ion
LC50 420
LC50 690
LC50 42,100
LC50 32, 900
SPECIES
Growth rate EC50
= 3.1? vol/vol
Reference
Mayer 4 Sanders, I97J
U.S. EPA, 1978
Birge, et al. 1978
Blrge, et al. 1978
Sanborn, et al. 1975
Birge, et al. 1978
Blrge, et al. 1978
Birge, et al . 1978
Birge, et al. 1978
Wilson, et al. 1978
-------
Table 6. (Continued)
IB
I
Species
Grass shrimp (larva),
Palaemonetes puglo
Grass shrimp (larva),
Palaemonetes puglo
Mud crab (larva),
RhIthropanopeus
harrlslI
Mud crab (larva),
Rh I thropanopeus
narrlslI
Chemical
dJ-2-ethylhexyl
phthalate
Dimethyl
phthaIate
Dimethyl
phthalate
dl-n-butyl
phthalate
Duration
Effect
Entire None on survival
larval and developmental
deveIopment ra te
Entire
larval
development
Entire
larval
development
Entire
larval
development
Significant
decrease in sur-
vival; Increased
Inter mo It and
developmental
periods
None on
development
None on
development
Result
(ug/ll
1,000
100,000
* Based on actual concentrations of dl-n-octyl phthalate accumulated
Lowest Freshwater Value: di-lsononyl phthalate = 420 ug/l
dl-2-ethylhexyl phthalate = 14-54 ug/l
dl-n-butyl phthalate - > 10,000 ug/l
dl-n-octyl phthalate = 690 ug/l
Lowest Saltwater Value: dimethyl phthalate = 100,000 ug/l
1,000
1,000
Reference
Laugh I in, et al. 1978
Laugh! In, et al. 1978
Laugh I In, et al. 1977
Laughlin, et al. 1977
-------
REFERENCES
Birge, W.J., et al. 1978. Effects of polychlorinated biphenyl compounds and
proposed PCB-replacement products on embryo-larval stages of fish and amphi-
bians. Research Report No. 118, University of Kentucky, Water Resources Re-
search Institute, Lexington, Kentucky.
Gledhill, W.E., et al. 1980. An environmental safety assessment of butyl
benzyl phthalate. Env. Sci. Techno!. 14: 301.
Laughlin, R.B., et al. 1977. Effects of Polychlorinated Biphenyls, Poly-
chlorinated Napthalenes, and Phthalate Esters on Larval Development of the Mud
Crab Rhithrppanopeus harrisii. In_: Pollutant Effects on Marine Organisms.
D.C. Heath Co., Lexington, Massachusetts, p. 95.
Laughlin, R.B., Jr., et al. 1978. The effects of three phthalate esters on
the larval development of the grass shrimp Palaemonetes pugio (Holthius).
Water, Air, Soil Pollut. 9: 323.
Mayer, F.L. 1976. Residue dynamics of di-2-ethylhexylphthalate in fathead
minnows (Pimephales promelas). Jour. Fish Res. Board Can. 33: 2610.
Mayer, F.L., et al. 1977. Collagen Metabolism in Fish Exposed to Organic
Chemicals. In: Recent Advances in Fish Toxicology, a Symposium. EPA 600/3-
77-085. U.S. Environ. Prot. Agency, Corvallis, Oregon, p. 31.
Mayer, F.L., Jr. and H.O. Sanders. 1973. Toxicology of phthalic acid esters
in aquatic organisms. Environ. Health Perspect. 3: 153.
8-20
-------
Mehrle, P.M. and F.L. Mayer. 1976. Di-2-ethylhexylphthalate: Residue Dynam-
ics and Biological Effects in Rainbow Trout and Fathead Minnows. _In: Trace
Substances in Environmental Health. University of Missouri Press, Columbia,
Missouri. p. 519.
Monsanto Industrial Chemicals Company. 1978. Acute toxicity of di-2-ethyl-
hexylphthalate (OEHP) to Daphnia magna. Report No. ES-SS-78-9, St. Louis,
Missouri.
Sanbom, J.R., et al. 1975. Plasticizers in the environment: The fate of
di-N-octyl phthalate (OOP) in two model ecosystems and uptake and metabolism
of OOP by aquatic organisms. Arch. Environ. Contam. Toxicol. 3: 244.
Sanders, H.O., et aT. 1973. Toxicity, residue dynamics, and reproductive ef-
fects of phthalate esters in aquatic invertebrates. Environ. Res. 6: 84.
Streufert, J.M. 1977. Some effects of two phthalic acid esters on the life
cycle of the midge (Chironomus plumosus). M.S. Thesis. Univ. of Missouri,
Columbia, Missouri.
U.S. EPA. 1978. In-depth studies on health and environmental impacts of
selected water pollutants. Contract No. 68-01-4646.
Wilson, W.B., et al. 1978. The toxicity of phthalates to the marine dino-
flagellate Gymnodinium breve. Bull. Environ. Contam. Toxicol. 20: 149.
B-21
-------
Mammalian Toxicology and Human Health Effects
INTRODUCTION
The annual production of phthalic acid esters in the United States in
1977 amounted to approximately 1.2 billion pounds. Since 1945, the cumula-
tive total production (up to 1972) of these esters reached a figure of 12.5
billion pounds (Peakall, 1975). On a worldwide scale, 3 to 4 billion pounds
are produced annually.
When the term "phthalate esters" is used, it indicates the ortho form of
benzenedicarboxylic acid. Two other isomeric forms of benzenedicarboxylic
acid esters are also produced. These include the meta form (or isothalate
esters) and the para form (or terephthalate esters). Both of these isomers
have a number of important commercial applications such as starting materi-
als for plastics and textiles. In this document, however, consideration
will be given only to the "ortho" esters.
Pthalic acid esters have a large number of commercial uses, the largest
being as plasticizers for specific plastics such as polyvinyl chloride.
Other uses for these esters include: defoaming agents in the production of
paper, in cosmetic products as a vehicle (primarily diethyl phthalate) for
perfumes, in lubricating oils, and in other industrial and consumer applica-
tions.
Dioctyl phthalate (includes di-2-ethylhexyl phthalate and other dioctyl
phthalates) accounts for approximately 42 percent of the esters produced in
this country, followed by diisodecyl phthalate. Dioctyl phthalate (OOP) and
di-2-ethylhexyl phthalate (DEHP) are often used synonymously even though it
should be clear that they are not the same, one being an isomer of the other.
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The extremely large production of phthalates and the variety of uses for
these esters have led to the presence of these esters in water sources,
food, consumer products, air (industrial settings, automobiles having vinyl
furnishings), and in medical devices such as tubings and blood bags. Esters
can thus enter the environment and biological species, including man,
through a variety of sources.
Therefore, man is exposed to phthalates from a variety of routes such
as: (1) ingestion from water, (2) ingestion from food, (3) inhalation,
(4) dermal, and (5) parenteral administration (via blood bags and tubes in
which the ester is extracted by a oarenteral solution including blood).
EXPOSURE
Inqestion from Water
In the early seventies, a great deal of attention began to focus on
chemical contaminants in surface water and adjacent ocean regions. One of
the first reports published on the presence of phthalic acid esters was pre-
sented by Corcoran (1973). He indicated that a level of approximately 0.6
ppm DEHP was present at the mouth of the Mississippi River. He further cal-
culated that approximately 350 million pounds of the ester enter the Gulf of
Mexico from the Mississippi River each year. As pointed out by Peakall
(1975), the 350 million pounds stated by Corcoran must be in error and may
be due to an error in the analytical procedure or to an abnormal local con-
centration. Corcoran also indicated the presence of DEHP (or its eouiva-
lent) in the Gulf near Pensacola, Florida and in the clear blue waters of
the Gulf Stream, but the levels of the esters were much less than at the
mouth of the Mississippi.
Hites (1973) studied chemical contaminants in the Charles and Merrimack
Rivers in Massachusetts. He reported that approximately 7 miles from the
C-2
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mouth of the Charles River the level of phthalate was 1.8 to 1.9 ppb. As
the water approached the mouth of the river, the level was reduced. For
example, three miles from the mouth, the level was 1.1 ppb while at one mile
from the mouth, the level ranged from 0.88 to 0.98 ppb.
A review of various EPA reports shows that surface waters do contain
phthalate esters in parts per billion, with the levels being higher at sites
close to industrial centers.
Ingestion from Food
A number of packaging materials and tubings used in the production of
foods and beverages are polyvinyl chloride contaminated with phthalic acid
esters, primarily OEHP. These esters migrate from the packaging to the food
or beverages. The extent of migration depends upon a number of factors such
as temperature, surface area contact, lipoidal nature of the food, and
length of contact. Peakall (1975) refers to reports on the migration of
plasticizers from tubings used in milk production. Extracted levels for the
dinonyl phthalate ester (in PVC tubing) were found to be 4.6 mg/100 ml/day
at 38°C and 7.0 mg/100 ml/day at 56°C. The rate for OEHP was 2.0 mg/100
ml/day at 38°C and 3.1 mg/100 ml/day at 56°C. The tubing was 1 meter in
length and 100 ml of milk was the extracting medium. Peakall suggests that
in actual practice approximately 40 mg of OEHP could be extracted over a 15
day period from tubings in contact with milk, but indicated that the actual
levels in milk are not known. A German report (Pfab, 1967) indicates that
cheese and lard placed experimentally in contact with two plastic films (one
containing dibutyl and the other dicyclohexyl phthalates) extracted less
than one percent of the esters after one month at 25°C. The concentra-
tions in the food were reported as less than 2 ppm.
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Food and Drug Administration (FDA) surveys indicate that several of the
phthalate esters are present °i.n food and fish which have had contact with
plastic packaging systems such as polyvinyl chloride (PVC). Some data on
the residue of the esters in Japanese foods have also been reported. Table
1, taken from the study by Tomita, et al. (1977) shows the amounts of sever-
al agents migrating to selected Japanese foods packaged in plastics, lami-
nated films, paper, and aluminum foil. As will be noted, levels above 600
ppm and even higher than 3,000 ppm of total phthalates migrated to certain
foods.
A bioconcentration factor (BCF) relates the concentration of a chemical
in aquatic animals to the concentration in the water in which they live.
The steady-state BCFs for a lipid-soluble compound in the tissues of various
aquatic animals seem to be proportional to the percent lipid in the tissue.
Thus the per capita ingestion of a lipid-soluble chemical can be estimated
from the per capita consumption of fish and shellfish, the weighted average
percent lipids of consumed fish and shellfish, and a steady-state BCF for
the chemical.
Data from a recent survey on fish and shellfish consumption in the
United States were analyzed by SRI International (U.S. EPA 1980). These
data were used to estimate that the per capita consumption of freshwater and
estuarine fish and shellfish in the United States is 6.5 g/day (Stephan,
1980). In addition, these data were used with data on the fat content of
the edible portion of the same species to estimate that the weighted average
percent lipids for consumed freshwater and estuarine fish and shellfish is
3.0 percent.
An average measured steady-state bioconcentration factor of about 330
was obtained for di-2-ethylhexyl phthalate using fathead minnows (Mayer,
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TABLE 1
Migration of Phthalic Acid Esters from Packaging Film to Foodstuffs*
Time after
Foodstuffs Manufacture
(months)
Tempura (frying) A 3
powder B 4
Instant cream A 14
soup B ?
C ?
o
tj-, Instant soybean 7
soup
Soft margarine 4
Fried potato A 1
cake B ?
C ?
Orange juice 1
Red ginger pickles ?
Table salt ?
Packaging Materials (ppm)
Materials**
Pl-1
Pl-L
P-A1-P1
P-A1-P1
P-A1-P1
P-P1
PI
P-PL
P-PL
P-PL
P-P1
PI
P-P1
DNBP
70.28
6.29
23.17
586.16
588.75
2.75
1.29
10.86
10,66
22.98
1.52
7.24
5.18
DEHP
3,675.0
2.30
1.35
58.92
58.93
1.85
1.44
385.85
1.28
11.80
0.74
2.75
2.58
Total
3,745.28
8.59
24.52
647.08
647.08
4.60
2.73
396.91
11.94
34.78
2.26
9.99
7.76
Foodstuffs loom]
DNBP
14.70
0.39
1.73
60.37
51.79
nd
nd
1.11
nd
1.21
0.35
nd
nd
DEHP Total
68.08
0.11
0.04-
2.15
3.01
nd
nd
0.05
nd
9.06
0.05
nd
nd
82.78
0.50
1.77
62.52
54.80
nd
nd
1.16
nd
10.27
0.40
nd
nd
*Source: Tomita, et al. 1977
**P1 indicates plastic, L indicates laminated film, P indicates paper. Al indicates aluminum foil.
-------
1976). Similar fathead minnows contained an average of 7.6 percent lipids
(Veith, 1980). An adjustment factor of 3.0/7.6 = 0.395 can be used to ad-
just the measured BCF from the 7.6 percent lipids of the fathead minnow to
the 3.0 percent lipids that is the weighted average for consumed fish and
shellfish. Thus, the weighted average bioconcentration factor for di-2-
ethylhexyl phthalate, and the edible portion of all freshwater and estuarine
aquatic organisms consumed by Americans is calculated to be 330 x 0.395 =
130.
Measured steady-state bioconcentration factors of 57, 117, and 663 were
obtained for dimethyl, diethyl, and butylbenzyl phthalates, respectively,
using bluegills (U.S. EPA 1980). Similar bluegills contained an average of
4.8 percent lipids (Johnson, 1980). An adjustment factor of 3,0/4.8 = 0.625
can be used to adjust the measured BCF from the 4.8 percent lipids of the
bluegill to the 3.0 percent lipids that is the weighted average for consumed
fish and shellfish. Thus, the weighted average bioconcentration factors for
dimethyl, diethyl, and butylbenzyl phthalates, and the edible portion of all
freshwater and estuarine organisms consumed by Americans are calculated to
be 36, 73, and 414, respectively.
No measured steady-state bioconcentration factor with an appropriate
percent lipids is available for dibutyl phthalate. However, log BCF is
nearly proportional to log P (Veith, et al. 1979). Thus, using values for
log P (Hansh and Leo, 1979) and the weighted average BCF values of 73 and
130 derived above for diethyl and di-2-€thylhexyl phthalates, respectively,
the weighted average bioconcentration factor for dibutyl phthlate and the
edible portion of all freshwater and estuarine aquatic organisms consumed by
Americans is estimated to be 89.
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Inhalation
This route may be a significant portal of entrance for esters of phtha-
11 c acid, at least to selected populations at risk. The presence of the
esters in air for relatively short periods of time most likely is due to the
incineration of PVC items. In closed spaces such as automobiles having PVC
furnishings, the ester can volatilize and the persons inside the vehicle
will inhale the vapors.
In closed rooms which have PVC tiles, levels of esters may reach 0.15 to
0.26 mg/m3 (Peakall, 1975). Mens'shikova (1971) reported the presence of
dibutyl phthalate (D8P) from ship ouarters furnished with PVC tile, decora-
tive laminated plastics and pavinols (assumed to be PVC plastics). He re-
ported that even after three years, the level of DBP in the air of the rooms
contained from 0 to 1.22 mg/m3 of the ester.
Milkov, et al. (1973) reported that vapors or aerosols of phthalate
esters ranged from 1.7 to 40 mg/m at one working site where mixing was
done, and a level of 10 to 66 mg/m at another working site in a company
manufacturing artificial leather and films of PVC.
American published reports regarding levels of esters in the working
environment are rare. Thus, insufficient data are available to judge what
levels of these esters are present in various working sites manufacturing
the esters or using the esters for consumer products.
It seems reasonable to assume that certain workers will be exposed to
the phthalic acid esters in the form of the vapor or as mists. Depending
upon the hygiene standard maintained, these workers could inhale sufficient
concentrations of the ester to lead to health problems.
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Dermal
The phthalate esters can be absorbed through the skin and this route may
thus become an important portal of entrance. Many cosmetic products may
contain small concentrations of the lower molecular weight phthalate esters
such as diethyl phthalate, and thus, application to the skin could introduce
the ester to humans through the skin. Because dimethyl phthalate is used as
a mosauito repellent, dermal absorption can occur. Swimming pools lined
with PVC could also release the phthalate esters to the water and, in turn,
swimmers would be exposed to very minute concentrations of the plasticizer
(phthalate esters) which could then be absorbed through the skin. As with
the other routes, lack of available data prevents even a very crude projec-
tion of the levels of esters which could enter man through the skin.
Because a number of medical devices such as blood bags, infusion con-
tainers, collection and administration tubings, and catheters are prepared
from plasticized (generally DEHP) polyvinyl chloride, a parenteral route of
entrance into a selected human population becomes a possibility. In fact,
it is possible that the parenteral route contributes the greatest Quantity
of the esters to selected groups under medical care in hospitals. These
medical devices have been introduced into medical practice since Walter
(1951) first introduced the polyvinyl chloride blood bag in 1950, and thus,
"many millions of persons have been exposed to phthalate esters by the
parenteral route."
The total number of renal hemodialyses performed each year in the United
States has reached close to six million. A single five-hour dialysis will
expose these patients to approximately 150 mg of DEHP. In open heart sur-
gery, extra corporeal pump oxygenators are used. Approximately 360,000 such
r Q
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operations are performed each year. Under these conditions, a patient may
be exposed to an average of 33 mg of OEHP during the surgery.
As early as 1960, a report appeared by Meyler, et al. (1960) that cer-
tain medically used PVC tubings released toxic ingredients to solutions
passed through them. Isolated heart experiments were used to detect toxic
ingredients released from PVC. Since these specific "toxic" tubings con-
tained an organotin stabilizer, the authors surmised that the toxic compo-
nent was the stabilizer and not the phthalate ester.
Braun and Kumrnel (1963) reported that PVC containers used for storage of
blood and transfusion solutions did release phthalate esters as well as
other additives to an extracting medium (water).
A report by Guess, et al. (1967) revealed that a number of American PVC
blood bags containing an anticoagulant solution (ACD) were contaminated by
the presence of small amounts of OEHP, 2-ethylhexanol, phthalic anhydride,
phthalic acid, and some unidentified chemicals.
Jaeger and Rubin (1970) reported the release of phthalate esters from
PVC blood bags and tubings, and further identified these plasticizers in
tissues and organs of two deceased patients who previously were transfused
with blood from PVC blood bags.
Hillman, et al. (1975) identified the presence of DEHP in neonatal tis-
sues after the insertion of umbilical catheters. It was interesting to note
that three infants who died of necrotizing enterocolitis had significantly
higher DEHP values in the gut than infants not having this disorder. There
was generally an increase in OEHP content of tissue if the specific patient
had also received blood products. Residue levels were measured in both
C-9
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heart and gastrointestinal tissues. The average level of DEHP in heart tis-
sue was 1.27 ug/g. In the gut of the three patients having died of gastro-
intestinal disorders, the levels ranged from 0.016 to 0.63 ug/g.
It is now well recognized that plasticized PVC medical devices will
release the plasticizers to tissue and to solutions in contact with the
object. Extraction of a plasticizer such as DEHP with water is extremely
small with the present PVC blood bags and infusion containers, but if lipoi-
dal solutions such as blood and blood fractions are used, the extent of re-
lease becomes significant.
The quantity of di-2-ethylhexyl phthalate released into stored blood at
4°C for 21 days ranges from 5 to 7 mg/100 ml (Jaeger and Rubin, 1972).
Kevy, et al. (1978) have done extensive studies on DEHP and found the
plasticizer to be extracted from PVC storage containers into blood and blood
components. A summary of some of their extract results is shown in Table 2.
Needham and Luzzi (1973) and Needham and Jones (1978) indicated that
when PVC infusion containers containing normal saline were agitated, DEHP
would occur in colloidal form in the saline. Even under this condition,
however, the total concentration of the colloidal particles came to 0.1 ppm
(Darby and Ausman, 1974). The presence of ethyl alcohol in the solution
will increase the level of DEHP in the solution. A 10 percent solution will
increase the DEHP content to 6 ppm, while a concentration of 40 percent will
increase the DEHP in the solution to 30 ppm (Corley, et al. 1977).
The total auantity of DEHP that a transfused patient may receive paren-
terally will, of course, depend upon the number of units of blood or blood
products administered to him. Patients undergoing chronic transfusions with
whole blood, packed cells, platelets, and plasma stored in PVC containers
C-10
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TABLE 2
Extraction Data of DEHP from PVC Containers*
1. Normal whole blood stored at 4°C contains 0.19 mg percent DEHP on
collection and 5.84 mg percent after 21 days of storage.
2. Cryoprecipitate which is prepared and stored at -30°C contains low levels
of DEHP (1.05 to 2.6 mg percent).
3. The level of DEHP in stored platelets maintained at 4°C and 22°C after 72
hours is 10.85 mg percent and 43.21 mg percent, respectively.
*Source: Kevy, et al. 1978
C-ll
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may receive a total of approximately 70 mg of DEHP. There are cases, how-
ever, when a patient may receive as many as 63 units of blood containing
approximately 600 mg of DEHP (Jaeger and Rubin, 1972).
PHARMACOKINETICS
Absorption
The phthalic acid esters and/or their metabolites are readily absorbed
from the intestinal tract, the intraperitoneal cavity, and the lungs. There
is also evidence indicating that these esters can be absorbed through the
skin. As will be pointed out, the vehicle can play an important role in the
absorption, distribution, and elimination of the ester.
Schulz and Rubin (1973) administered orally to rats 14C-OEHP in corn
oil and found that approximately 13 percent of the administered dose was
found in the organic solvent extracts of urine, feces, and contents of the
large intestine. The urine contained about 62 percent in water extracts.
Daniel and Bratt (1974) injected a single oral dose of 14C-OEHP in rats
and found 42 percent and 57 percent of the dose in the urine and feces, re-
spectively, in seven days. They also pointed out that a significant amount
of the dose is excreted in bile~. In studies by Wall in, et al. (1974) rats
were orally administered ring or side chain-labeled DEHP. Twenty-four hours
after the dose was given, approximately 50 percent of the recovered radio-
activity was found in the feces and in the gastrointestinal tract contents.
The remaining radioactive substance was recovered in the urine. The authors
also indicated that "a portion of the radioactivity recovered from the feces
undoubtedly had been absorbed but returned to the gut in the bile."
Lake, et al. (1975) have suggested that orally administered phthalic
acid esters are absorbed in the gut primarily as monoesters. Wallen, et al.
(1974) however, found from their studies that a significant amount of orally
C-12
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administered DEHP is absorbed in the gastrointestinal tract as the intact
compound. From the present, data, it appears clear that the diester phtha-
lates can be hydrolyzed to the monoester in the gut and thus be absorbed as
the monoester. Further studies are needed to clarify the ratio of intact
diester to monoester which would be absorbed in the gut under various condi-
tions in several species of animals.
Information on the absorption of the phthalic acid esters in man is lim-
ited. As early as 1945, however, Shaffer, et al. (1945) reported that a
single oral dose of 10 g OEHP in a human subject was recovered as a phtha-
late eauivalent in the urine after 24 hours. The amount recovered was 4.5
percent of the original dose. In another subject, 5 g DEHP was taken orally
and 2.0 percent of the original dose (as phthalate eauivalent) was found in
the urine 24 hours later. Tomita, et al. (1977) reported the presence of
phthalate esters in the blood of individuals having ingested food which had
been in contact with flexible plastics having the phthalic acid esters.
DEHP and di-n-butyl phthalate (DN8P) levels detected in the blood after
meals were much higher than prior to eating the foods in the plastic packag-
ing system. In 13 individuals who were included in the study, DEHP and DNBP
in the blood ranged from 0.13 to 0.35 ppm when compared to an average value
of 0.02 ppm prior to the meals.
Dillingham and Pesh-Imam detected nine percent in urine 24 hours after
labeled DEHP had been applied to rabbit skin. After 48 hours, the levels in
the urine had increased to 14 percent and within 72 hours the radioactivity
had increased to 16 to 20 percent of the originally administered dose.
Distribution
Absorbed esters of phthalic acid esters (or their metabolites) distrib-
ute auite rapidly to various organs and tissues both in animals and humans.
C-13
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Again, it must be kept in mind that, depending upon the route and the physi-
cal form of the ester (true solution, colloid, emulsion), the distribution
of the esters (metabolites) can vary. Jaeger and Rubin (1970) studied the
distribution of DEHP in human tissues of two deceased patients having had
large volumes of blood (stored in PVC blood bags) transfused into them.
They detected the presence of OEHP in the spleen, liver, lung, and abdominal
fat with concentrations ranging from 0.025 mg/g in spleen to 0.270 mg/g in
abdominal fat.
Radio-labeled OEHP (emulsified in oleic acid) administered intravenously
(i.v.) as a single dose was found to disappear rapidly from the blood and
approximately 60 to 70 percent of the total dose was detected in the liver
and lungs within two hours of the dose (Daniel and Bratt, 1974). In studies
in which rats were maintained on diets containing DEHP, there was a progres-
sive increase in the amount of the compound in the liver and abdominal fat
of the animals but within a short time a steady state concentration was
achieved (Daniel and Bratt, 1974).
Waddell, et al. (1977) examined the distribution of 14C-DEHP (serum
solubilized) after a single i.v. injection in rats using whole body auto-
radiography techniaues. Results from the study revealed that a rapid accu-
mulation of radioactivity in the kidney and the liver had occurred followed
by rapid excretion into urine, bile, and intestine. No accumulation of the
compound was found (up to 168 hours after the injection) in the spleen and
lung, but significant radioactivity was detected in the lumen of the intes-
tine which the authors surmised occurred because of the secretion of the
compound by the liver into the bile.
Tanaka, et al. (1975) administered 14C-DEHP solubilized in Tween 80^
orally to groups of rats. The concentrations in the liver and kidney
C-14
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reached •> maximt in *he first two to six hours. Peak blood levels of
the compound occurred about six hours after administration. Intravenous
administration of labeled DEHP as a dispersion prepared by sonification of
DEHP in saline led to 70 to 80 percent of the original dose deposited in the
liver'after the first hour. After two hours, the radioactivity had declined
to 50 percent and only 0.17 percent radioactivity was found in the liver at
the end of the seventh day. The intestine (after oral and i.v. administra-
tion) revealed a relatively high level of radioactivity but not to the same
extent as the liver. On the other hand, the testicles and brain appeared to
have little affinity for the compound regardless of the route of administra-
tion. Other organs and tissues also showed low levels of radioactivity
after 24 hours of oral dosing.
Dillingham and Pesh-Imam injected i.v. a single dose of labeled DEHP in
mice and found that after seven days the highest specific activity resided
in the lungs, with lesser amounts in the brain, fat, heart, and blood
(Autian, 1973). These investigators did not find preferential deposition of
OEHP (as radioactivity) in fatty tissue. Application of labeled diethyl
phthalate to the skin of rabbits resulted in detection of the compound in
the lungs, heart, liver, kidney, gonads and spleen after three days. The
compound (or its metabolite) was also detected in the brain but, surprising-
ly, no radioactivity was detected on the skin or subdermal fatty tissue at
the site of application.
With the current information on distribution of the phthalate esters, it
can be concluded that the esters are rapidly distributed to various organs
and tissues with no apparent accumulation. Yet it is now well-recognized
that the general population and patients having received large-volume blood
or blood products may have residues of phthalate esters or metabolites in
C-15
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tissues and organs. A study by Jacobson, et al. (1977), in which nonhuman
primates were transfused with blood containing DEHP following a procedure of
treatment common to humans, revealed the presence of DEHP (or metabolites)
in trace amounts even up to 14 months post-transfusion. As pointed out by
Daniel and Bratt, (1974), there probably is a steady state concentration
which is reached after which the esters (or metabolites) are then rapidly
eliminated from the organs or tissues through various routes, thus prevent-
ing significant accumulation over long periods of exposure.
Metabolism
Albro, et al. (1973) have identified the metabolites of DEHP after oral
feeding to rats. These authors conclude that the first step in the metabo-
lism is the conversion of the diester to monoester (mono-2-ethylhexyl phtha-
late). By u>- and (
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that a single oral dose of labeled DEHP was practically all excreted in
urine and feces within a four day period, leaving less than 0.1 percent of
the radioactivity in the organs and tissues. Rats pretreated with DEHP for
6 and 13 days also showed a similar elimination rate upon the administration
of labeled DEHP. Excretion into bile also appears to be a significant route
of excretion increasing the content of DEHP (or metabolites) in the intes-
tine.
Schulz and Rubin (1973) intravenously administered labeled DEHP to
groups of rats and then monitored the radioactivity in blood versus time.
They noted a bi-phasic curve when the data were plotted as log DEHP vs.
time. The initial slope led to a half-life in blood of nine minutes while
the second slope gave a half-life of 22 minutes. Within one hour, 8 percent
of the total injected DEHP was found in water-soluble metabolites, primarily
in the liver, intestinal contents and urine. Twenty-four hours after injec-
tion, 54.6 percent of the initial dose was recovered as water-soluble metab-
olites primarily in the intestinal tract, excreted feces, and urine. Only
20.5 percent was recovered in organic extractable form.
Dillingham and Pesh-Imam studied the excretion in the urine of mice of
labeled DEHP administered i.p. (as pure ester) and i.v. (as saturated saline
solution), (Autian, 1973). They noted that 68 percent and 63 percent, re-
spectively, of the total initial dose was excreted in seven days.
Tanaka, et al. (1975) reported about 80 percent of the original labeled
DEHP given orally or by i.v. to rats was excreted in the urine and feces in
five to seven days. These authors also pointed out that, upon a single oral
administration of DEHP, the intact diester could not be identified in the
urine. On the other hand, repeated oral administration of 500 mg/kg in rats
C-18
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for 20 days revealed the presence of intact OEHP in the urine. They con-
cluded that "repeated administration of DEHP may lead to its accumulation in
the body until a steady state is reached between the rates of absorption and
elimination." After steady state is reached, DEHP as the unchanged mole-
cule, would appear in the urine.
As Thomas, et al. (1978) have expressed in their review article on bio-
logical effects of OEHP, pharmacokinetic data in animals and humans support
the thesis that OEHP is absorbed from the gastrointestinal tract and widely
distributed to various tissues following either the oral or i.v. routes of
administration. OEHP is then rapidly metabolized to a number of derivatives
of mono-2-ethylhexyl phthalate which are, in turn, excreted mainly in the
urine. The half-life of elimination from tissues and the body is short.
EFFECTS
Acute, Subacute, and Chronic Toxicity
One of the first comprehensive reviews on the toxicity of phthalate
esters was presented by Autian in 1973. A much more detailed review of the
phthalate esters was given by Peakall in 1975 and the most recent one on
this subject was published by Thomas, et al. in 1978. The potential health
threats of phthalic acid esters in the early seventies led to a national
conference on the subject in 1972. The papers presented at this meeting
were published in the January 1973 issue of Environmental Health Perspec-
tives. As will become evident, most of the detailed toxicological studies
have centered primarily on DEHP since this specific ester accounts for ap-
proximately 40 percent of the phthalates which are used commercially.
From the accumulated data on acute toxicity in animals, the phthalate
esters may be considered as having a rather low order of toxicity. It is
now thought that the toxic effect of the esters is most likely due to one of
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the metabolites in particular to the monoester. This appears to be the case
for OEHP since this ester has been studied more extensively than the others.
Table 3 is taken from Autian's 1973 review and lists the ID™ of the es-
ters. Oral acute toxicity for the lower molecular weight esters is greater
in animals than for the higher molecular weight esters such as OEHP. Other
routes of administration such as i.p. and dermal do not significantly in-
crease the acute toxicity (Autian, 1973).
The toxicity of DEHP by the i.v. route is ouite important since, as has
been indicated previously, PVC administration devices will leach the plasti-
cizer into blood and lipo-protein-containing solutions. Since DEHP has a
very limited solubility in water, other means of administering the agent in
experimental animals have been used to study the toxic effects when adminis-
tered i.v. Preparation of emulsions or dispersion of DEHP in various vehi-
cles may induce toxic responses when injected i.v. which may not occur when
DEHP is solubilized by having the ester migrate from PVC into blood. Stud-
ies by Stern, et al. (1977) have indicated that the pharmacokinetic pattern
for DEHP will be different depending upon the vehicle which is used and they
make the suggestion that i.v. studies should be performed on the extracted
DEHP which will take place when the blood product is placeed in contact with
a PVC device. Since DEHP will have a limited solubility in blood and blood
products, the total dose given to animals will be relatively small and, in
general, no acute toxicity would be expected. Rubin (1976), however, has
suggested the possibility of "shocked lungs" when DEHP is administered i.v.
and has presented experimental evidence in rats to support this contention.
This is discussed in a subsequent section of this report. The low volatili-
ty of most of the esters precludes them from presenting an acute toxic re-
sponse by inhalation. Generally, at least for the higher molecular weight
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TABLE 3
Acute Toxicity of Phthalate Esters:
in Animals*
Compound
Dimethyl
phthalate
Di ethyl
phthalate
Dimethoxyethyl
phthalate
Diallyl
phthalate
Di butyl
phthalate
Diisobutyl
phthalate
Butyl carbobutoxy
methyl phthalate
Animal
Mouse
Mouse
Mouse
Rat
Rat
Guinea pig
Rabbit
Mouse
Mouse
Rat
Rabbit
Mouse
Mouse
Rat
Rat
Guinea pig
Guinea pig
Mouse
Rat
Rabbit
Rabbit
Mouse
Rat
Rat
Rabbit
Mouse
Mouse
Rat
Guinea pig
Rat
Rat
Route
oral
i.p.
i.p.
oral
i.p.
oral
dermal
i.p.
i.p.
i.p.
oral
oral
i .p.
oral
i.p.
oral
dermal
i.p.
oral
oral
dermal
i.p.
i.p.
i .m.
dermal
oral
i.p.
i.p.
dermal
oral
i .p.
LD50
g/Kg
7.2
3.6
1.58
2.4
3.38a
2.4
10. Oa
2.8
2.8
5.06a
1.0
3.2-6.4
2.51
4.4
3.7
1.6-3.2
10.0
0.7
1.7
1.7
3.4a
4.0
3.05a
8.0
20. Oa
12.8
4.50
3.75a
10. Oa
14. 6a
6.89
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TABLE 3 (cont.)
Compound
Dihexyl
phthalate
Dioctyl
phthalate
Di-2-ethyhexyl
phthalate
Butylbenzyl
phthalate
Dicapryl
phthalate
Oinonyl
phthalate
Dibutyl (diethylene
glycol bisphthalate)
Dialkyl
phthalate
Animal
Rat
Rabbit
Mouse
Rat
Guinea pig
Mouse
Rat
Rat
Rabbit
Guinea pig
Mouse
Mouse
Rat
Mouse
Mouse
Rat
Rat
Mouse
Rat
Route
oral
dermal
oral
i .p.
dermal
i .p.
oral
i.p.
oral
dermal
i .p.
i.p.
oral
oral
i.p.
oral
i.p.
oral
i .p.
LD50
g/Kg
30.0
20. Oa
13.0
50. Oa
5.0a
14.2
26.0
50. Oa
34.0
10.0
3.16
14.2
2.00
11.2
11.2
11.2
11.2
20.00
20.00
*Source: Autian, 1973
in ml/kg
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phthalic acid esters, only through heating will there be sufficient vapor
concentration to carry out an adeauate inhalation study.
Even though the phthalate esters have been in commercial production for
nearly 50 years, relatively few long-term toxicity studies appear in the
literature. As would be expected, subacute (or subchronic) studies are more
plentiful but even these are few when one considers the large production of
these agents every year. Perhaps the meager toxicological data can be at-
tributed to the long use of these esters with relatively few episodes of ill
effects among the general population. Also, it is possible that a number of
these esters have been studied in more toxicological detail by industry
without the results appearing in published form. A general indication of
long-term toxicity of phthalate esters can be seen in Table 4 in which
Krauskopf (1973) has summarized the maximum no-effect dose for several
esters.
Dimethyl Phthalate: Dimethyl phthalate is used as an effective mosquito
repellent. In human experience, few toxic effects from this ester have been
noted. Two-year feeding studies in female rats by Draize, et al. (1948) at
levels of 2 and 8 percent in the diet produced only a minor growth effect at
the 4 and 8 percent levels. At the 8 percent level, some indication of
nephritic involvement was detected. Dose levels less than 8 percent showed
no such effect. A 90-day study in which the ester was applied to the skin
of rabbits led to an LD5Q of greater than 4 ml/kg. The ester does not
produce primary irritation on the skin nor has it been found to act as a
sensitizing agent.
Diethyl Phthalate: This ester has been used as a plasticizer for cellu-
lose materials and as a perfume carrier. Nearly 50 years ago, Smith (1924)
reported that rats could tolerate up to 0.5 percent of their body weight of
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TABLE 4
Calculated Allowable Daily Intake (ADI) for Various
Phthalate Esters*
Ester
Di-2-ethylhexyl
Dibutyl
Diisonyl
Heptyl nonyl
Species
Rat
Rat
Dog
Rat
Doq
Rat
Guinea pig
Dog
Rat
Rat
Rat
Dog
Rat
Mouse
Period
Days
365
730
98
90
98
365
365
365
365
450
91
91
90
90
Maximum
Mo-Effect Level
(mg/kg/day)
400
80
100
200
100
60-200
60
60
350-110
4.3
150
37
60
60
*Source: Krauskopf, 1973
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this ester without death occurring. Rabbits could be fed 3 ml/kg/day with-
out significant toxic response (Blickensdorfer and Templeton, 1930).
In a two year study (Food Research Laboratories, Inc., 1955) groups of
30 rats (15 of each sex) were fed either 0.5, 2.5, or 5.0 percent diethyl
phthalate in the diet. No effects were observed at levels of 0.5 or 2.5
percent. Diethyl phthalate at 5.0 percent resulted in a small but signifi-
cant decrease in the growth rate of the rats without any effect on food con-
sumption. Thus, 5.0 percent diethyl phthalate appeared to affect the effi-
ciency of food conversion. Also in this study, dogs were fed diethyl phtha-
late at levels of 0.5, 1.5, 2.0, and 2.5 percent for one year. Problems
were encountered with palatability of diethyl phthalate in the diet. As a
result, the dogs received varying exposures to diethyl phthalate before each
dog attained stabilization at the highest dietary level that could be toler-
ated. Accordingly, three dogs were maintained at 0.5 percent, one each at
1.5 and 2.0 percent, and three at the 2.5 percent level. The average weekly
intakes of diethyl phthalate were computed and found to be 0.8, 2.4, 3.5,
and 4.4 g/kg/week in order corresponding to increasing dietary level. No
effects were noted at any of these levels.
Diethyl phthalate does not act as a primary irritant when applied to the
skin nor has it induced allergic responses in humans who have contact with
it. Heated vapors may produce slight irritancy in mucous membranes of
the nasal passages and may also irritate the upper respiratory tract.
Even though diethyl phthalate is not generally used as a plasticizer in
PVC tubings, Neergaard, et al. (1975) reported that this ester was present
in tubings used in hemodialysis eouipment and that the use of these tubings
led to hepatitis in several patients. When other tubings, presumably with-
out diethyl phthalate, were used the hepatitis did not occur. It seems un-
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likely that the ester was responsible for the hepatitis and the cause may
have been related to another additive in the tubing.
Dibutyl Phthalate: Smith (1953) studied the effects of feeding dibutyl
phthalate to groups of rats. At concentrations of 0.01, 0.05, and 0.25 per-
cent of dibutyl phthalate in food, no adverse effects were noted after one
year. When the dose level was increased to 1.25 percent, approximately half
of the animals died in the first week but the remaining animals grew normal-
ly as compared to the untreated controls.
Spasovski (1964) conducted an inhalation study lasting 93 days during
which mice were exposed for six hours a day to different concentrations of
the ester. The concentrations ranged from 0.017 to 0.42 mg/m . Unfortu-
nately, during the study, the same animals received various exposure concen-
trations rather than specific concentrations for the whole time period and
thus interpretation of the results is difficult even though Spasovski pro-
posed a permissible standard concentration (PSC) of 1 mg/m3. Dvoskin, et
al. (1961) exposed groups of rats to 0.2 and 0.4 mg/m3 for 2.5 months.
Some weight loss was noted and an increase of gamma globulin was reported
for the animals receiving the higher exposure during the fourth and sixth
weeks of the experiments. The same group of animals also demonstrated
alterations in the phagocytic activity of neutrophils after one month; these
returned to normal. It is difficult to conclude from this study the signif-
icance of the results in regard to the toxic potential of dibutyl phthalate
when inhaled.
A much more detailed study on the inhalation of dibutyl phthalate has
been reported by Men'shikova (1971). Rats were exposed continuously for 93
days at chamber concentrations of 0.098, 0.256 and 0.98 mg/m3. No behav-
ioral changes were noted nor any weight loss discerned. The important find-
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ing was that gamma globulin was increased and appeared to be dose related.
In humans, Men'shikova (1971) found an olfactory threshold value ranging
from 0.26 to 1.47 mg/m . Atmospheric concentrations of 0.12 and 0.15
mg/m resulted in abnormal encephalographic responses in the three human
subjects in the study. When the level was reduced to 0.093 mg/m3 no con-
ditioned reflex was noted. Men'sikova recommends a PSC value of 0.1 mg/m3.
Carter, et al. (1977) described a study on dibutyl phthalate and the
resultant testicular atrophy which occurred. In the study, the ester was
dissolved in corn oil and administered orally (by intubation) for a period
of time. The dose administered was 2,000 mg/kg while control animals re-
ceived corn oil in a volume of 5 ml/kg. The initial effect noted was a pro-
gressive reduction in weight of the testes. In 14 days, the reduction
amounted to 60 to 70 percent of the original weight. Since there was also a
decrease in body weight, the authors used "relative testes weight" and found
that even in this manner of reporting there was still a significant loss
(testes weight). Histopathological methods on testes tissue demonstrated
morphological damage. Further investigations by these authors revealed that
the ester apparently influenced zinc metabolism with an increase in the
excretion of zinc in urine. It was visualized that after oral administra-
tion, dibutyl phthalate is metabolized by nonspecific esterases in the gas-
trointestinal tract to the monobutyl phthalate prior to absorption into the
bloodstream. Results from the various experiments have led the authors to
suggest that the monoester or another metabolite of dibutyl phthalate may be
acting as a chelating agent by removing the zinc from the testes. The defi-
ciency of zinc in testes tissues is, according to the authors, the causative
factor leading to the atrophized organ.
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Milkov, et al. (1973) reported in 1969 that a group of esters in an in-
dustrial environment produced various degrees of toxic polyneuritis. These
investigators studied 147 persons (87 women and 60 men), the majority of
whom were not more than 40 years old. These industrial workers were exposed
primarily to dibutyl phthalate but other esters apparently were also present
but in much lower concentrations. These included dioctyl, diisooctyl and
benzyl butyl phthalates. Also, in some instances there were small amounts
of sebacates, adipates, and tricresyl phosphate.
Until more occupational studies are performed, the report by Milkov, et
al. (1973) must be taken with some reservation because of the presence of
other chemical agents such as tricresyl phosphate, an agent known for induc-
ing polyneuritis.
Dibutyl (diethylene glycol bisphthalate) (DQGB): Hall, et al. (1966)
studied the toxicity of ODGB. They used a commercial sample which also con-
tained 15 percent dibutyl phthalate and 5 percent (diethylene glycol) phtha-
late. The oral ID™ of this product in rats was found to be greater than
11.2 g/kg and the i.p. LD5Q approximately 11.2 g/kg. A 12-week toxicity
study was conducted on the product using rats as the test animals. Diets in
different groups of rats contained 0, 0.25, 1.0, and 2.5 percent of the
product, respectively. Over the period of the study, there was a marked
reduction of growth in the treated animals as compared with the control
group. Also evident were enlargements of the liver and heart at the 1.0 and
2.5 percent levels in male rats and enlarged brain in both male and female
animals. At the 2.5 percent level, oxaluria and hematuria were found in
both sexes, the oxaluria being assumed to be a direct conseauence of the j_n_
vivo liberation of diethylene glycol (a known producer of oxalate stones in
the bladder).
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Ethylphthalyl ethyl glycolate (EPEG): Hodge, et al. (1953) conducted a
chronic oral toxicity study with EPEG in rats for two years and dogs for one
year. Groups of 50 rats (25 of each sex) were fed either 0, 0.05, 0.5, or
5.0 percent EPEG in the diet. No effects were observed at levels of 0.05 or
0.5 percent. EPEG at 5.0 percent resulted in decreased growth and survival.
Kidney damage was also observed at 5.0 percent EPEG. This consisted of mot-
tled granular kidneys, sometimes swollen and pale yellow. The pelvis of the
kidney was usually dilated, and frequently a hard, stag-horn calculus, a
fine sand, or both were seen. In the study performed with dogs, dose levels
of 0, 0.01, 0.05, and 0.25 gm/kg/day were used with two male dogs per dose
level. No effects were observed at any of these levels.
Butylphthalyl butyl glycolate (BPBG): Information on the chronic toxic-
ity of BPBG comes from two unpublished reports to the FDA.
Sovler, et al. (1950) conducted a two-year feeding study in rats (10 per
exposure group) at levels of 0.02, 0.2, or 2.0 percent BPBG in the diet.
Sovler, et al. reported that there was no evidence of toxicity or retarda-
tion of growth at any of the levels of BPBG. In the second study (Hazleton
Laboratories, 1950) groups of 20 rats were fed average daily doses of either
4, 40, or 389 mg/kg/day in order corresponding to 0.02, 0.2, and 2.0 percent
BPBG in the dietary levels of BPBG.
Butyl benzyl phthalate: Mallette and Von Hamrn (1952) administered both
orally (1.8 g/kg) and i.p. (4 g/kg) butyl benzyl phthalate to groups of
rats. Animals died after four to eight days and histopathological studies
demonstrated toxic splenitis and degeneration of central nervous system tis-
sue with congestive encephalopathy. Further, myelin degeneration and glial
proliferation were reported.
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Dialkyl 79 phthalate: This product contains a mixture of phthalate
esters of alcohols having chain lengths of seven to nine carbons. In a
90-day feeding study in rats by Gaunt, et al. (1968) no demonstrable adverse
effects were noted at diet levels of 0.125 percent, but at the 0.5 and 1.0
percent levels, increased liver weights were observed even though histo-
pathological changes were not seen. The authors concluded that a 60 kg
adult could ingest 36 mg/day without any apparent harm.
Di-2-ethylhexyl phthalate (DEHP): As has been indicated a number of
times, this ester is the most used phthalate and for this reason more toxi-
cological data are available on it than any of the other esters. It should
be remembered that DEHP is often used synonymously with the dioctyl phtha-
late and, even though they are isomers, they have slightly different biolog-
ical properties. The acute oral toxicity for rodents ranges from 14.2 to
greater than 50 g/kg. Dermal absorption occurs but in rabbits approximately
25 ml/kg must be applied to the skin to cause death.
In 1945, Shaffer, et al. (1945) reported a 90-day subacute toxicity
study in rats. Groups of animals were given in feed 0.375, 0.75, 1.5, and
3.0 percent of the ester which approximates daily intakes of 0.2, 0.4, 0.9,
and 1.9 g DEHP/kg per rat in the four treated groups while the fifth group
served as a control (no phthalate). At the three higher levels (0.75, 1.5,
3.0), a slight decrease in growth was noted when compared to the control
animals. At the 1.5 and 3.0 percent doses, tubular atrophy and degeneration
in the testes were observed. No deaths occurred in any of the treated ani-
mals while blood cell counts, hemoglobin concentrations and differential
white cell counts remained normal. The authors concluded that no adverse
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effect from oral administration would occur at approximately 0.2 q/kg/day or
less while only a slight retardation in growth may occur when the dose is
increased to 0.4 g/kg/day.
Carpenter, et al. (1953) conducted a study on chronic oral toxicity of
DEHP using rats, guinea pigs, and dogs. In the rat study, parental (P,)
generation rats received daily diets containing 0.04, 0.13, and 0.4 percent
of OEHP for a maximum period of two years. In addition, a group of filial
generation (F^ rats were given in feed 0.4 percent of DEHP for one year.
Control groups of rats were maintained on the same basic diet without the
ester. The investigators examined the following signs and symptoms of tox-
icity: mortality, life expectancy, body weight, food consumption, liver and
kidney weiqhts, micropathological changes, neoplasm, hematology and fer-
tility.
Over the two-year period for the PI group and over a one-year period
for the Fj group, a number of deaths occurred. However, these deaths were
not attributed to the ester since they were also noted in the control ani-
mals.
The mean liver and kidney weights, as percentage of body weights, were
found to be increased over those of the controls in both the initial group
(Pj) and their offspring (Fj) which had received the diet containing 0.4
percent DEHP. The results were statistically significant. Histopathologi-
cal examination of the liver and kidney tissues of treated animals did not
reveal statistically significant differences from organs of control animals.
The authors did suggest that even though pathological changes in the two
organs of treated groups were not different from control animals, the in-
crease in size of the organs may indicate a toxic response. Results from
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comparisons of life expectancy, body weight, food consumption, neoplasia,
hematology and fertility in the treated animals were found not to differ
significantly from controls.
In another study by the same investigators (Carpenter, et al. 1953),
groups of guinea pigs were administered in diet 0.04 and 0.13 percent DEHP
for one year. Similar criteria, with the exception of hematology and fer-
tility, as used in the rat study were employed. Liver weights, as percent-
age of body weights, were found to be statistically higher in the treated
groups than in the control animals. The authors pointed out that the effect
was not related to the concentrations since both treated groups appeared to
be about the same in regard to liver weight. The other parameters studied
were found not to be significantly different from control animals. A "no
effect" dose for DEHP in guinea pigs (for one year) was estimated to be 0.06
g/kg/day.
A one-year study was also reported by Carpenter, et al. (1953). In this
study, dogs were administered capsules with 0.013 ml/kg/day DEHP, five days
a week, for the first 19 doses and then 0.06 ml/kg/day until 240 doses had
been administered. No statistically significant adverse effects were seen.
The authors concluded that a "no effect" dose in dogs would be approximatley
0.06 g/kg/day.
Harris, et al. (1956) published a paper which, in effect, confirmed the
results of Carpenter, et al. (1953). A chronic oral toxicity study in male
and female rats was conducted in which groups of animals received in their
feed 0, 0.1 and 0.5 percent DEHP. At various time periods, rats were sacri-
ficed and food consumption records, body weight, and liver, testes, kidneys,
lungs, brain, stomach, heart and spleen weights examined. Histopathological
studies were also conducted on selected tissues and organs. The study was
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terminated after 24 months. Significant increases in liver and kidney
weights were noted at the 0.5 percent dose level for the three and six-month
sacrifices. At the one and two year periods, no real differences in the
liver and kidney weights were apparent in any of the groups, but the authors
point out that this may have been due to the small number of rats remaining
after these longer periods. No unusual pathology was noted in the tissues
and organs prepared for microscopic examination which could be attributed to
the ester. Slight body weight reduction was seen at the 0.4 and 0.5 percent
dose. Food consumption was decreased at the 0.5 percent level when compared
to the control animals.
In a dog study, Harris, et al. (1956) reported a mild toxic effect with-
in three months when a dog was administered 5 g/kg/day of DEHP but not with
0.1 g/kg/day. The small number of dogs in this study (two) and relatively
short period of study (14 weeks) do not permit a valid conclusion to be made
of the chronic effects of DEHP on dogs. However, this data considered with
the data of Carpenter, et al. (1953) suggests that a no-effect dose in dogs
is approximately 0.1 g/kg/day.
Lawrence, et al. (1975) studied the subchronic toxicity of a number of
phthalate esters to determine the chronic LDj-n by the i.p. route. Groups
of male mice were administered a range of doses for each of the esters, five
days a week, and an apparent LD5Q calculated for that week. This dosing
schedule was continued until two criteria were met: (1) mice were injected
for at least ten weeks, and (2) the apparent LD5Q remained constant for
three consecutive weeks. DEHP and OOP were included in the list of esters
studied. The first week, the LD50 for DEHP was 38.35 ml/kg and 67.18
ml/kg for OOP. The second week, the LD5Q was reduced to 6.40 ml/kg for
DEHP and 25.51 ml/kg for OOP. By the end of the 12th week, the LD5Q was
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reduced to 3.09 ml/kg for OEHP and to 1.37 ml/kg for OOP. A cumulative tox-
icity factor was calculated for each of the esters (acute LD50/chronic
LDrn) and for DEHP this value was 27.99 (indicating that the toxicity had
increased by this factor). A similar calculation for OOP came to 21.74.
The other esters had cumulative toxicity factors ranging from 2.05 to 4.01,
indicating that cumulative toxicity was only minimal over the time period
the animals were studied. The implication of the high cumulative toxicity
factors for both DEHP and OOP is not clear and the reasons for these re-
sults, when compared to the other esters, are presently not explainable. It
is possible to speculate that very high exposure doses prevent the body from
eliminating the compound and metabolites to the same degree as occurs when
repeatedly lower doses are administered. It is also not known if oral doses
would have led to the same or similar results, since this type of adminis-
tration was not done in the study by Lawrence, et al. (1975).
Earlier studies by Shaffer, et al. (1945) Carpenter, et al. (1953) and
Harris, et al. (1956), demonstrated the low chronic toxicity of DEHP but
they also noted that at the higher daily doses kidney and liver enlargement
occurred. These investigators, however, could not find light microscopic
evidence of injury to these organs using histopathological methods. The
enlargement of an organ such as the liver may not necessarily indicate that
a toxic event has occurred, as suggested by Golberg (1966).
In studies by Lake, et al. (1975), rats were orally dosed with DEHP in
corn oil at a concentration of 2,000 mg/kg/day for periods of 4, 7, 14, and
21 days. Control animals received 0.5 ml/100 g body weight of the vehicle.
The investigators noted relative liver weight increased progressively during
the treatment to 215 percent of the controls at the end of 21 days. Liver
homogenates were prepared for each time period and the following biochemical
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activities and/or levels determined (for each of the time periods): succi-
nate dehydrogenase, aniline 4-hydroxylase, biphenyl 4-hydroxylase, glucose-
6-phosphatase, cytochrome P-450, protein contents, and alcohol dehydrogen-
ase. Alcohol dehydrogenase activity and microsomal protein and cytochrome
P-450 contents increased markedly initially but then decreased during the
time of treatment. On the other hand, microsomal glucose-6-phosphatase,
aniline 4-hydroxylase, and mitochondrial succinate dehydrogenase activity
decreased significantly. Electron microscopy of liver tissue of treated
animals demonstrated changes in hepatocytes. At the end of seven days,
there was an increase in microbodies and there also appeared to be a dila-
tion of the smooth endoplasmic reticulum and swelling of the mitochondria.
Lake, et al. (1975) studied the monoester and found that liver changes
in treated rats closely resembled those produced by DEHP. They concluded
that in general the toxic effects of DEHP are due to the metabolite, mono-2-
ethylhexyl phthalate.
Daniel and Bratt (1974) fed dietary concentrations of 1,000 and 5,000
ppm of C-DEHP to groups of female rats for 35 and 49 days, respectively.
Two animals from each group were sacrificed at various intervals and the
heart, brain, liver, and abdominal fat removed for radiochemical analysis.
Remaining animals were returned to a normal diet and sacrificed at intervals
during the subsequent two to three weeks and tissues orepared for analysis.
At the 5,000 ppm level, liver weight relative to total body weight increased
progressively during the first week to a value approximately 50 percent
above the control and remained constant in the remaining time period. Elec-
tron microscopy of liver tissue revealed only a slight increase in the
amount of smooth endoplasmic reticulum. Returning animals to a normal diet
resulted in liver weight returning to normal. There was no apparent change
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in liver weight in those animals receiving the 1,000 ppm OEHP. Additional
studies by these authors did not reveal the accumulation of DEHP in body
organ tissues.
Nikonorow, et al. (1973) reported that a daily dose level of 0.35 per-
cent (in feed) of OEHP caused a decrease in body weight of rats after 12
months. In other chronic studies on DEHP, livers of treated animals were
significantly larger than livers from control animals not receiving DEHP.
Kevy, et al. (1978) studied the toxic effects of DEHP solubilized in
monkey blood or blood products by storing the animal blood (or blood prod-
uct) in PVC blood bags. These products were then transfused into the ani-
mals for time periods ranging from six months to one year. This dosing pro-
gram attempted to mimic actual transfusion levels expected in selected pa-
tients reauiring large-volume blood or blood products. The total concentra-
tion of DEHP received by the monkeys ranged from 6.6 mg/kg to 33 mg/kg.
Liver damage was noted by several sensitive tests (hepato-splenic ratio
using an isotopic techniaue and BSP kinetic compartmental analyses) as well
as routine light microscopy of liver tissue. Even up to 32 months after the
last transfusion, liver changes persisted. DEHP was also found in liver
tissue in treated animals many months after the last transfusion. The work
of Kevy and his associates has significance since DEHP can enter man through
various PVC medical devices. Mild-to-moderate hepatic toxicity may occur
depending upon the dose, the freauency of exposure, and the health status of
the patient.
Biochemical studies on rat blood and liver at 21 days after i.p. injec-
tion of 5 ml/kg DEHP on days one, five and ten produced the following re-
sults: a decrease in the activity of succinic dehydrogenase and an increase
in alkaline phosphatase activity in the liver; serum enzyme values were not
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altered. This study was conducted by Srivastava, et al. (1975) who pointed
out that DEHP may also play a role in interfering with energy metabolism of
the cell.
Though it is recognized that different routes and dosage forms will
alter the pharmacokinetic disposition of compounds, DEHP from several dif-
ferent routes (oral, i.p., i.v.) can produce hepatotoxic responses depending
upon the specific dose and the freouency of exposure.
Seth, et al. (1977) administered i.p. 5 ml/kg of OEHP (undiluted) to 10
male and 20 female rats on days 1, 5, and 10. On the 22nd day of the study,
all animals were sacrificed and one testis or ovary was removed and retained
for enzymatic studies. A control group of rats received an eoual volume of
saline. Results of the study demonstrated that the scrotums in all animals
were enlarged but no gross abnormality was discerned. Succinic dehydrogen-
ase (SDH) and adenosine triphosphatase (ATPase) activities were significant-
ly reduced, while that of s-glucuronidase was increased in both organs of
the test animals. Histopathologic examination of the testes of the animals
revealed degenerated tubules showing marked vacuolization of the cytoplasma
of spermatogonial cells and eccentric nuclei. No apparent alterations (his-
topathologic) were noted in the ovaries of the OEHP treated rats.
Carter, et al. (1977) alluded to an unpublished study on DEHP in which
rats were fed various dose levels of the ester for 90 days. At a daily
level of 0.2 percent, DEHP produced testicular injury. When the level of
DEHP was increased to 1.0 percent, testicular injury was noted in two weeks.
The authors further state that DEHP and dibutyl phthalate have about the
same potency in causing testicular atrophy in rats. Even though mention was
made that other esters of phthalic acid were studied, no data were present-
ed. Thus, the reader may assume that these other esters did not have the
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same toxic properties to testes as either DEHP or the dibuty] ester. It
seems possible that DEHP, like dibuty1 phthalate, may affect zinc metabolism
in the testes which, in turn, may be the causative factor in bringing about
atrophy of the organ.
In a series of papers, Bell, et al. (1976, 1978) have demonstrated that
feeding rats DEHP can have an effect upon lipid metabolism including inhibi-
tion of heoatic sterologenesis, inhibition of fatty acid oxidation by heart
.-
mitochondria, stimulation of fatty acid oxidation by hepatic mitochondria,
and an ability to modify the pattern of circulating plasma lipoproteins. In
several of the studies, rabbits and pigs were also used and led to the con-
clusion that the response of mammalian tissues to phthalate esters is varia-
ble depending upon the species. The toxic implications of alteration in
lipid metabolism to man are presently obscure.
The toxic properties of DEHP are most likely related to the formation of
the monoester (in the gut or liver) and/or to other metabolites produced in
the body. Studies by Lake, et al. (1975) demonstrated that neither phthalic
acid nor 2-ethylhexanol reproduced the toxic effect of DEHP, suggesting that
the metabolites must play the major factor in producing a toxic response.
It also appears that man, rats, baboons, and ferrets may handle DEHP as well
as other esters in a similar manner (Lake, et al. 1977).
Synergism and/or Antagonism
There are no data available on the synerqism or antagonism of phthalate
esters.
Teratoqenicity
Singh, et al. (1975) included eight phthalic acid esters in a rat tera-
togenic study. The esters included the following: dimethyl, dimethoxy-
ethyl, diethyl, dibutyl, diisobutyl, butyl carbobutoxymethyl, dioctyl and
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di-2-ethylhexyl phthalates. For all the esters, except two, the dose admin-
istered i.p. to pregnant female rats was 1/10, 1/5, and 1/3 the acute
LD5Q. For these esters, the doses ranged from a low of 0.305 ml/kg for
dibutyl phthalate to a high of 2.296 ml/kg for butyl carbobutoxymethyl
phthalate. Di-2-ethyhexyl phthalate and dioctyl phthalate were given at
doses of 5 and 10 ml/kg because of their very low acute toxicity. Control
groups included: untreated rats, rats treated with 10 mg/kg of distilled
water, rats treated with 10 ml/kg of normal saline and rats treated with 10
ml/kg and 5 ml/kg of cottonseed oil. All treatments took place on days 5,
10, and 15 of gestation. On the 20th day, all the rats were sacrificed and
the uterine horns and ovaries were surgically exposed to permit counting and
recording of the number of corpora lutea, resorption sites, and viable and
dead fetuses. Additionally, both viable and nonviable fetuses were excised,
weighed, and examined for gross malformation. From 1/3 to 1/2 of the fetus-
es, using those which showed no gross malformation when possible, were pre-
pared as transparent specimens to permit visualization of skeletal deformi-
ties.
All of the esters produced gross or skeletal abnormalities which were
dose related. The most common gross abnormalities in the treated animals
were absence of tail anophthalmia, twisted hands and legs, and hematomas.
Skeletal abnormalities included elongated and fused ribs (bilateral and uni-
lateral), absence of tail bones, abnormal or incomplete skull bones, and
incomplete or missing leg bones. Dead fetuses were found in the groups
treated with dimethyl, dimethoxyethyl, and diisobutyl phthalates. The most
embryotoxic agent in the series was dimethoxyethyl phthalate. Each of the
esters also reduced the weight of the fetuses when compared to the controls.
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Even at the high dose levels (5 and 10 ml/kg), di-2-ethylhexyl and dioctyl
phthalates had the least adverse effects on embryo fetus development.
Since the study by Singh, et al. (1972) was carried out i.p., results
should not be extrapolated to possible teratogenic effects if the compounds
have been administered orally or by other routes.
In another study by Peters and Cook (1973), pregnant rats were adminis-
tered i.p. 4 ml/kg DEHP on days three, six and nine of gestation. At this
dose level, implantation was prevented in four of five rats. When the dose
was reduced to 2 ml/kg, a similar response was noted in three of five rats.
These authors also noted adverse effects on parturition in dams treated with
OEHP such as excessive bleeding, incomplete expulsion of fetuses and mater-
nal deaths. Teratogenic studies on dibutyl and dimethyl phthalates were
also conducted by these authors, but the adverse effects were less than
those observed for the DEHP-treated rats. It was interesting to note that
adverse effects prior to gestation day six were primarily on implantation,
while after this day the effect was primarily on parturition.
In another study by Singh, et al. (1975), rats were injected i.p. with
labeled di-2-ethylhexyl phthalate and diethyl phthalate. The results demon-
strated that these phthalates could pass through the placenta! barrier sug-
gesting that the embryo-fetal toxicity and teratogenesis of the phthalic
acid esters could be the result of the direct effect of the compound (or its
metabolites) upon developing embryonic tissue.
Bower, et al. (1970), studied the effects of eight commercial phthalate
esters in chick embryos. They found that dibutyoxyethyl phthalate, di-2-
methoxyethyl phthalates, and octyl isodecyl phthalate produced damage to the
central nervous system of the developing chick embryo when compared to con-
trol embryos receiving an oil and to an untreated group.
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In a study reported by Nikonorow, et al. (1973), pregnant rats were
administered orally 0.34 and 1.70 g/kg/day of DEHP during the gestation
period. Another series of rats received orally 0.120 and 0.600 g/kg/day of
dibutyl phthalate. Olive oil was used as a control and administered in a
similar manner as the esters to a group of rats. There was a statistically
significant reduction in fetus weight at both dose levels for DEHP but only
at the higher dose level for the dibutyl phthalate. The number of resorp-
tions were noted for DEHP at both dose levels but only at the higher dose
level for dibutyl phthalate. No detectable differences were observed in the
number of sternum ossification foci, development of the bones at the base of
the skull, paws of the front and hind legs, and rib fusion in fetuses when
compared to the control animals.
Since the Quantity of phthalate esters ingested by humans on a daily
basis is extremely small as compared to the doses used in the previous stud-
ies, it seems remote that teratogenic effects would be produced in humans.
Further studies in which the esters are administered orally to pregnant
females should, however, be carried out to verify this assumption.
Mutagenicity
Studies of the effect of phthalic acid esters on genetic changes in ani-
mals are not adeauate to conclude if one or more of these compounds presents
a threat to animals and man. One of the few studies published on this topic
is by Singh, et al. (1974). These authors included DEHP and dimethoxyethyl
phthalate (DMEP) in a study on the mutagenic and antifertility effects in
mice. The experiment followed the general procedure used in conducting the
dominant lethal assay for mutagens. A group of ten males were injected i.p.
with each compound at three doses. For the DEHP, the doses were 1/3 (12.8
ml/kg), 1/2 (19.2 ml/kg), and 2/3 (25.6 ml/kg) of the LD5(J. A similar
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dose pattern was used for the DMEP or 1/3 (1.19 ml/kg), 1/2 (1.78 ml/kg) and
2/3 (2.38 ml/kg) of the LD50.
Each group of male mice was injected with the doses shown above and,
immediately following injection, each male was caged with two virgin adult
female mice. Each week for 12 weeks, two new virgin females replaced the
previous week's female mice.
Results of the study indicated that at the high dose of both esters a
distinct reduction in the incidence of pregnancies occurred. Fewer effects
were noted at the lower dose levels. DEHP appeared to have a more persis-
tent effect over the time period studied than DMEP. Both esters produced
some degree of dose and time-dependent antifertility and mutagenic effect.
Early fetal deaths occurred indicating the potential mutagenic effects of
these compounds. The increase in early fetal deaths was not large, however,
it was above the values for the control animals.
Rubin, et al. (1979) included a number of phthalate esters in an Ames
mutagenic assay. The esters included: dimethyl, diethyl, dibutyl, mono-2-
ethylhexyl, di-2-ethylhexyl, and butyl benzyl phthalate as well as phthalate
acid. Positive responses were found for the dimethyl and diethyl phtha-
lates. The remaining compounds were found to be non-mutagenic under test
conditions.
Studies by Turner, et al. (1974), showed the DEHP did not produce genet-
ic damage in lymphocytes but did inhibit mitosis and growth. It is clear
that more studies on the mutagenic effects must be conducted before a defi-
nite conclusion can be made concerning the risk of a population exposed to
the phthalate esters. The antifertility effect appears to be much stronger
and the Question which still needs to be answered is what effects would low-
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er doses have upon males repeatedly exposed to these esters. Epidemiologi-
cal evidence on this subject is lacking, and thus human risks cannot accu-
rately be portrayed.
Carcinogencity
A recent report by Rubin, et al. (1979), alluded to under Mutagenicity
in which an in vitro mutagenic assay was conducted on a group of phthalate
esters (dimethyl, diethyl, dibutyl, mono-2-ethylhexyl, di-2-ethyhexyl, and
butyl benzyl phthalates) and on phthalic acid showed that both dimethyl and
diethyl phthalates produced a positive response suggesting but not proving
that these compounds may have a cancer liability. The National Cancer In-
stitute is currently conducting bioassays on butyl benzyl phthalate (feed),
diethyl hexyl phthalate (feed), and diallyl phthalate (gavage). The results
of these bioassays will be reviewed when they are published.
Other Biological Effects
Cellular Toxicity: In recent years, a number of jji vitro tests have
become useful in assessing the toxicity of chemicals. Even though the re-
sults may not always be extrapolated to animals or humans, the proper _in_
vitro system can generate very useful data which can assist in determining
the toxic conseouences of a chemical. Tissue and organ culture methods are
now widely used in toxicity testing methods.
Nematollahi, et al. (1977) synthesized and purified a number of phthalic
acid esters and then included them in a toxicity screening program using two
cell lines (chick embryo and L-cells). The esters, as solids or liauids,
were placed on the surface of agar which overlaid the cells. A vital dye
was also included in the cells. For the solids, 20 mg of the ester were
placed on the surface while for the liauids, 35 mg of the ester were placed
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on a paper disk which was previously placed on the agar. After 24 hours of
incubation, the cells were examined for cytotoxicity. Table 5 includes the
results of the screening tests. In the same table are the results from a
mouse toxicity test. Three mice were injected i.p. at a concentration level
of 5 moles/kg in either cottonseed oil or castor oil, depending upon the
solubility of the specific compound. As will be seen from the table, the
lower molecular weight esters were cytotoxic and lethal to mice. Several of
the highest molecular weight esters also demonstrated some signs of toxicity.
Jacobson, et al. (1974), found that solubilized DEHP in serum inhibited
cell growth (normal diploid fibroblasts established from skin) in tissue
culture experiments. A concentration of 0.18 mM, which is eauivalent to
that in 21-day-old whole blood stored at 4°C, inhibited cell growth by 50
percent. A 20 percent reduction in cell growth occurred when the OEHP con-
centration was reduced to 0.10 mM which is comparable to the concentration
found in whole blood stored at 4°C for 14 days.
In another tissue culture study Jones, et al. (1975) reported the IDrg
(concentration required to imnbit cell growth by 50 percent) on a number of
phthalic acid esters. The*ID,-Q values are shown in Table 6. As will be
noted from the table, IDcn for DEHP came to 70 yM. In comparing this
ID™ with the one reported by Jacobson, et al. (1974) (0.18 mM), it should
be remembered that the Jacobson group reported the concentration they added
to the culture medium, whereas Jones, et al. (1975), indicatmd the actual
solubility in the medium. The 70 pM solubility concentration would be ap-
proximately 0.05 mM which is in line with the Jacobson value considering
that slightly different techniques were employed. The most cytotoxic ester
in the series was butyl glycolyl butyl phthalate.
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TABLE 5
Results of the Toxicity Evaluation of Phthalate Esters
on the Mammalian Cell Cultures and Mice*
Phthalates
iso-C-H7
n-C5Hn
Cyc1o-C5Hg
n-C6H13
Cyclo-C6Hu
n-C7H15
Cyclo-C7H13
Cyclo-C8H15
n-C9Hlg
Chick
Alkyl Group Embryo L-Cells Mice
Cells
CH3
n~C3H7
Note: In tissue culture test: + indicates cytotoxic; - indi-
cates noncytotoxic; ^ indicates Questionable results.
In mouse test: + indicates 2 or 3 deaths; - indicates
no deaths; ^_ indicates only one death.
*Source: Nematollahi, et al. 1967
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TABLE 6
1050 Values for a Series of Phthalate Esters
Using WI-38 Cells*
Agent
(Phthalate)
Di-n -butyl
Di-iso-butyl
Dimethoxyethyl
Butyl glycol butyl
Di-n-octyl
Di-2-ethylhexyl
Molecular
Weight
278
278
282
336
391
391
ID50
yM
135
85
3,500
12
170
70
Solubility
(mole/1)
0.008
Very low
0.040
Very low
Very low
Very low
*Source: Jones, et al. 1975
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The ~0,.g do group of phthalate esters has been reported for
mouse fibroblasts in cell culture (Autian, 1973). These values are included
in Table 7. It is interesting to note that the most cytotoxic agent in the
series was DEHP, an agent having a very low order of acute toxicity in ani-
mals and man. As can be seen from the table, the toxicity of these com-
pounds, in general, increased as the molecular weight increased.
A report by Oillingham and Autian (1973), indicates that dimethyoxyethyl
phthalate is much more toxic to mouse fibroblast cells undergoing signifi-
cant rates of cell division than nonreplicating cells. This observation
suggests that any tissue which undergoes periodic increases in protein turn-
over related to changes in cell division rate and metabolic activity (pro-
tein synthesis) may increase the susceptibility of these cells to the toxic
effects of phthalic esters. Thus, it is possible that the teratogenic and
embryotoxic effects of several of the esters reported in rats may be due to
the fact that differentiating embryonic tissues have periodic major changes
in cell division rates and metabolic activity in contrast to somatic cells
which have a much lower rate of cell division and metabolism of the somatic
tissue.
Kasuya (1974) cultured cerebella from newborn rats and tested three
phthalate esters (dimethyl, diethyl and dibutyl phthalates). Various con-
centrations of each of the esters were dissolved in calf serum and then add-
ed to the cells. The overall toxicity to the cells was in the following
order: D8P>DEP>OMP. As will be noted, the toxicity of the three esters
increased with molecular weight similar to cell culture results reported by
Dillingham and Autian (1973).
At a concentration of 4 yg/ml in tissue culture media, DEHP produced
complete cessation of beating chick embryo heart cells maintained in tissue
C-47
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TABLE 7
of a Group of Phthalic Acid Esters in Tissue Culture
(Mouse Fibroblasts)*
Ester
Dimethyl
Diethyl
Oi butyl
Dimethoxyethyl
Di-2-ethylhexyl
Molecular
Weight
194
222
278
282
390
Water Sol.
(mole/1)
0.0263
0.0048
0.008
0.0400
0.0004
ID50
0.007
0.003
0.0001
0.0084
0.00005
*Source: Autian, 1973
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culture (Rubin and Jaeger, 1973). Up to 98 to 99 percent of the cells were
found to be dead within a 24-hour period. This result, along with the other
tissue culture reports, reinforces that DEHP is highly toxic at the cellular
level.
Blood Components/Lungs/Heart: In the past there has been concern that
DEHP, when extracted from medical devices such as blood bags and tubings,
might have a deleterious effect upon blood components and also lead to the
syndrome referred to as "shocked lungs." OEHP, solubilized with a surfac-
tant and injected i.v. in rats, produced lung involvement and death. Stern,
et al. (1977) have stressed the importance of the physical form of DEHP when
injected i.v.: the naturally solubilized DEHP showing a "nontoxic" effect
while DEHP solubilized with a surfactant produced a toxic effect.
Rubin (1975) reported that DEHP, solubilized with a surfactant and in-
jected i.v. in rats, produced a biexponential disappearance of the DEHP from
blood with half-lives of 3.5 and 35 minutes. A naturally solubilized DEHP,
on the other hand, has a monoexponential disappearance with a half-life of
19 minutes. In humans, Rubin (1975) found that the half-life of naturally
solubilized DEHP led to a monoexponential rate with a mean half-life of 28
minutes. Rats administered the surfactant solubilized DEHP showed death and
lung involvement similar to the shocked lung syndrome (Rubin, 1975).
Hypotensive rats, in which DEHP is added to the animal's own blood and
then transfused back into the rat, produced hemorrhagic lungs in each of the
six rats used in the experiment (Rubin, 1976). Control rats, treated in a
similar manner but not receiving any DEHP, did not demonstrate the toxic
lungs.
Berman, et al. (1977) conducted studies in which rats were administered
blood or blood components, previously in contact with PVC strips, to detect
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the effect OEHP (extracted from the plastic) would have on lung tissue.
ACO-preserved rat blood was stored in glass vials alone or in the presence
of sterile plastic strips. One set of plastic strips was also enriched with
34 percent OEHP. After storage for two weeks, 0.5 ml of blood were adminis-
tered i.v. to groups of rats in the following forms: as whole blood, as
whole blood minus platelets and buffy coat, as platelet-rich plasma, as
platelet-poor plasma. Additional groups of rats received CPO-preserved rat
or human blood after storage in glass alone or in glass containing PVC
strips and/or PVC enriched with DEHP. Concentration of DEHP in whole blood
in contact with PVC was 81.5 ug/ml and 90.2 ug/ml for the blood in contact
with PVC enriched with OEHP.
Evans Blue was used as an indicator to detect the permeability of ex-
cised lung tissue. Animals given ACD-preserved blood which had contact with
PVC demonstrated an increased permeability when compared to control animals.
Administration of platelet-rich and platelet-poor plasma showed no signifi-
cant increase in lung permeability. CPD-preserved blood in contact with the
plastic strips showed an increased permeability which was greater than the
CPO blood used as controls but not as great as the permeability shown by the
ACD-preserved blood. Histopathologic examinations of lungs having received
blood in contact with PVC and PVC enriched with OEHP showed variable degrees
of septal thickening, perivascular edema and perivascular accumulation of
mononuclear cells when compared to lungs of control rats. The authors sug-
gest that blood-plastic contact during storage may adversely affect blood
and also the effects may be in part due to accumulation of DEHP in red
cells. It has also been found that PVC infusion containers, if agitated,
will produce liauid particles of DEHP which, in turn, can be administered to
humans (Needham and Luzzi, 1973). Depending upon the size-freauency of
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these particles and the concentration of DEHP released to the solution, pos-
sible toxic effects may result even though human experience has not yet
indicated that adverse effects have occurred.
Vessman and Rietz (1978) have reported the presence of mono-2-ethylhexyl
phthalate (hydrolysis product of DEHP) in blood plasma stored in PVC blood
bags. Ten blood bags with plasma were removed from storage (-20.C) and the
monoester was found to range from 4 to 56 yg/ml. Eight of the plasma sam-
ples were then transferred to glass bottles and stored at room temperature.
After two weeks of storage the monoester contents had increased to values
between 27 and 79 yg/ml. Fractionated protein albumin also contained the
monoesters in amounts from less than 3 to 290 ug/g. The authors suggest
that the conversion of DEHP in plasma is due to some enzymatic activity tak-
ing place in the product. They indicate that when measuring DEHP content of
blood and blood products stored in PVC bags, attention should also be given
to determining the monoester content, thereby gaining a true picture of
phthalate content.
Sleeping Time: Sleeping time experiments were reported by Rubin and
Jaeger (1973) who studied the effect of DEHP and butyl glycolyl butyl phtha-
late. These esters were also emulsified with acacia and injected at 250
mg/kg and 500 mg/kg dose levels. After 30 minutes, hexobarbital solution
was administered i.p. A significant increase in sleeping time was produced
by DEHP at both dose levels, while only the higher dose of butyl glycolyl
butyl phthalate produced a longer sleeping time than the control animals.
Rats were also employed by the authors in a similar sleeping time experiment
with the results being similar but the magnitude less than with the mice.
Rubin and Jaeger (1973) conducted additional experiments and concluded that
the increase in hexobarbital sleeping time was not due to an increase in CNS
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sensitivity to hexobarbital nor an alteration in rate of hexobarbital metab-
olism by the liver, but to the effect of DEHP in the distribution of hexo-
barbital into various organs.
Swinyard, et al. (1976) also found an increase in hexobarbital sleeping
time from DEHP. It was interesting to note that olive oil also produced an
increased sleeping time similar to DEHP. These authors concluded that the
effect of DEHP was nonspecific due to the physical characteristic of the
ester which enlarged the lipophilic reservoir for hexobarbital rather than
to a pharmacological property of the compound.
Daniel and Bratt (1974) noted that hexobarbital sleeping time (in rats)
was increased when OEHP was used at a dose of 600 mg/kg of emulsified agent.
When rats were given orally five successive daily doses of DEHP (500/kg)
hexobarbital sleeping time was decreased.
From the information available, it is clear that DEHP prolongs the
sleeping time of short-acting barbiturates. In the instance of acute stud-
ies, the cause of the prolongation of sleeping time may, in fact, be due to
nonspecific factors, probably to the lipophilic reservoir mechanism advocat-
ed by Swinyard, et al. (1976). On the other hand, repeated pretreatments
with DEHP may have an effect upon the liver and enzyme systems. Since liver
involvement has been noted by several investigators in subacute toxicity
studies in rats and monkeys, the DEHP may, in these cases, be producing a
specific toxicological effect.
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CRITERION FORMULATION
Existing Guidelines and Standards
The Threshold Limit Value (TLV) for dimethyl, dibutyl and di-2-ethyl-
hexyl phthalate esters established by the American Conference of Government-
al and Industrial Hygienists (ACGIH) is 5 mg/m .
The Food and Drug Administration (FDA) has approved the use of a number
of phthalate esters in food packaging materials. Prior to 1959 (before
enactment of the food additive amendment), FDA approved five esters. These
are: diethyl phthalate, diisobutyl phthalate, ethyl phthalyl ethyl glycol-
ate, diisooctyl phthalate and di-2-ethylhexyl phthalate. Since then, 19
additional phthalates used in packaging material for foods of high water
content have also been approved. More specific uses and restrictions of
phthalic esters are set forth by FDA in its regulations.
Current Levels of Exposure
Lack of sufficient data prevents an accurate assessment of levels of
exposure of man and animals to phthalate esters. Is is now, however, well
known that man is exposed to these esters through a number of routes such as
industrial sites in which the esters are manufactured or used. Phthalate
esters may also reach man through indirect means such as inhalation of the
esters inside vehicles containing PVC products from foods and from water.
Direct injection i.v. of specific phthalate esters can also occur when PVC
blood bags and tubings are used to transfuse blood and blood products to
man. The ubiquitous nature of the phthalate ester is apparent since tissues
of deceased persons have revealed the presence of phthalic acid esters, even
though the individuals were not apparently exposed to these esters.
Even though it is well established that workers in occupations in which
phthalate esters are used are exposed to various levels of phthalate esters
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and thus can absorb these esters through inhalation or through dermal ab-
sorption, the lack of sufficent data precludes establishing what are the
levels of exposure. Dermal absorption of the low molecular weight esters
such as dimethyl phthalate (mosouito repellent) and diethyl phthalate (in
cosmetic products) probably is also occurring but the Quantity absorbed
through the skin is not known.
A survey was conducted by the Bureau of Foods (FDA) in 1974 to determine
if phthalate esters were entering the food supply through the processing,
packaging, handling and transportation chain. In the study, ten basic and
stable food products were analyzed for the presence of these esters. Con-
clusions reached in the report are presented here:
1. The freouency and levels of phthalate esters reported as well
as the possible cumulative intake of phthalates in baked beans
in cans or jars, canned whole kernel corn, margarine, cereals,
eggs, bread, corn meal, meat, milk, and cheese do not pose a
hazard to the consumer.
2. DEHP was the ester most frequently detected in the food com-
modities. Dibutyl phthalate, dicyclohexyl phthalate and
butylphthalylbutyl glycolate were found in comparatively few
samples. Diisoctyl and diisodecyl phthalates, although looked
for, were not detected.
3. Phthalate ester contamination was found in a higher proportion
of milk and cheese samples than in other foods. However, the
findings are uncertain.
In the above survey, the highest levels of phthalate esters were present
in margarine (13.7 and 56.3 ppm on fat basis). In cheese, the highest lev-
els of esters were 22.8 and 24.9 pom for DNBP and 35 ppm for DEHP but most
cheese samples contained less than 5 ppm of phthalates.
In a published study by Tomita, et al. (1977), information is presented
dealing with phthalate (DEHP and DNBP) residues in various commercial food-
stuffs in Japan. They concluded that foods packaged in plastic films with
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printing are a greater source of contamination to the product with the es-
ters than if the foods were in plastic bottles. They also noted that per-
sons had significantly higher levels of the esters after meals from foods
packaged in the film. Extremely high levels of the two esters (combined)
were found in tempura powder stored for eight months (up to 454 ppm). The
residue level of the esters from plastic films containing the plasticizers,
as would be expected, migrated to fatty foods or fatty-like foods to a
greater extent than to foods having low fat content. The authors included
in their conclusion the following: "The daily intake of PAEs (phthalic acid
esters) from present foodstuffs may not exceed the ADI of DNBP and DEHP but
an effort to reduce the PAE levels in foodstuffs should be continuously
made."
The Bureau of Foods (FDA) in another survey on fish from a number of
locations in the U.S. noted that the highest level of DEHP (7.1 ppm) was
present in shark (smooth, hound). In most other instances, the fish which
were studied were free of the esters.
Patients receiving repeated transfusions with whole blood, packed cells,
platelets and plasma stored in PVC may receive up to 70 mg of DEHP and, in
some instances, the auantity even exceeds 500 mg. Hemodialysis patients may
receive up to 150 mg of DEHP.
Special Groups at Risk
Two groups are at risk in regard to phthalic acid esters. These are
workers in the industrial environment in which the phthalates are manufac-
tured or used and patients receiving chronic transfusion of blood and blood
products stored in PVC blood bags.
C-55
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Basis and Derivation of Criterion
From the available-information, the phthalic acid esters have not been
found to be carcinogenic in animals or man. At high doses when injected
i.p., the esters can act as teratogenic agents and possibly as mutagenic
agents in rats. These esters also have an effect upon gonads in rats. Evi-
dence is also at hand to show that the esters may bring about biochemical
and pathological changes in the liver of rats when repeatedly administered
orally or by i.p. When solubilized in blood components, DEHP has demon-
strated liver involvement when these products have been repeatedly adminis-
tered i.v. to monkeys. Inhalation studies in rats and man suggest that cer-
tain phthalates may be responsible for neurological disorders, but these
results need further verification since other nonphthalate esters may also
have been present leading to the problems.
Since a number of phthalate esters are in the environment or may be
present in water, it was thought appropriate to review chronic toxicity data
in which well established chronic toxicity data for these esters were re-
ported to establish an acceptable daily intake (ADI). In Table 8 can be
found a summary of the studies chosen for the purpose of determining an ADI.
The table includes those ohthalate esters for which at least one reasonable
adeouate chronic ingestion toxicity study was available. Only in the case
of di-2-ethylhexyl phthalate was a choice between studies involved. The
Carpenter, et al. (1953) study was chosen because it was considered the most
representative.
The no-effect level from this study was supported by a majority of the
other chronic studies. In calculating the ADI, an uncertainty factor of 100
was used based on the National Academy of Sciences (NAS, 1977) guidelines.
C-56
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TABLE 8
Calculated Allowable Daily Intake In Water and Fish
for Various Phthalate Esters*
1.
2.
3.
4.
5.
6.
Ester
Dimethyl
Diethyl
Dibutyl
Di-2-ethyl-
hexyl
Ethylphthalyl
ethyl glycolate
Butylphthalyl
butyl glycolate
Species
Rat
Rat
Rat
Rat, Guinea
Pig, Dog
Rat
Rat
Weeks
104
104
52
104, 52. 52
104
104
No Effect Dosea
(mg/g/day)
1,000
1,250
125
60
250
1.000
ADI
(mg/day)
700
875
88
42
175
700
Water Quality
Fb Criterion
(mg/1)
36 313
73 350
89 34
130 15
NE
NE
Reference
Draize, et al.
1948
Food Research
Lab Inc., 1955
Smith, 1953
Carpenter,
et al. 1953
Hodge, et al.
1953
Solver, et al.
1950; Hazel ton
Labs.. 1950
*Source: Shibko, 1974
aWheri only a concentration in the diet was available, it was assumed that a rat consumes 5% of its body weight per day, on the average.
^Bioconccntration factor
-------
This safety factor is justified because all of the animal studies provide
sufficient data on chronic (greater than one year) exposure.
For the sake of establishing water duality criteria, it is assumed that
on the average a person ingests 2 liters of water and 6.5 grams of fish.
The amount of water ingested is approximately 100 times greater than the
amount of fish consumed. Since fish may biomagnify the esters to various
degrees, a biomagnification factor (F) is used in the calculation. Biocon-
centration factors for dimethyl, diethyl, dibutyl and di-2-ethylhexyl esters
were derived by the U.S. EPA ecological laboratories, Duluth, Minnesota (see
Ingestion from Food section).
Due to lack of data, bioconcentration factors could not be derived for
dicyclohexyl, methyl phthalyl ethyl glycolate, ethyl phthalyl ethyl glycol-
ate, and butyl phthalyl ethyl glycolate.
The eauation for calculating an acceptable amount of ester in water
based on ingestion of 2 liters of water and 6.5 g fish is:
(2 1) X + (0.0065 x F) X = ADI
where 21=2 liters of drinking water consumed
0.0065 = amount of fish consumed daily in kg
F = bioconcentration factor
ADI = Allowable Daily intake (mg/day for 70 kg person)
X = Water Quality criterion
For example, consider that the ADI for dimethyl phthalate is 700 mq/day
and the bioconcentration factor is 36, the above eauation can be solved as
follows:
2(X) + (0.0065 x 36) (X) = 700
2X + (0.23)X = 700
2.23X = 700
X = 313 (or ~ 310 mg/1)
C-58
-------
Thus, the recommended water duality criterion is 313 mg/1 for dimethyl
phthalate.
Similar calculations were made for each of the esters and are presented
below:
Oiethyl Phthalate
2(X) + (0,0065 x 73) (X) = 875
2X + G.*7X = 875
2.47X = 875
X = 354 mg/1 (or - 350 mg/1)
Dibutyl Phthalate
2(X) + (0,0065 x 89) (X) - 88
2X + 0.578X - 88
2.578X = 88
X = 34.1 mg/1 (or - 34 mg/1)
Di-2-ethylhexyl Phthalate
2(X) + (0.0065 x 130) (X) = 42
2X + Q.345X = 42
2.845X = 42
X = 14.8 mg/1 (or ~ 15 mq/1)
Thus, the recommended water quality criteria for four phthalate esters
are:
dimethyl = 313 mg/1
diethyl = 350 mg/1
dibutyl = 34 mq/1
di-2-ethylhexyl = 15 mq/1
(see Table 8),
:-5§
-------
It seems clear that exposure from the water route presents no real risk
to the population in regard to the phthalate esters. Reported levels of
phthalate esters in U.S. surface waters have only been in the ppb range, at
approximately 1 to 2 ug/1 (see Ingestion from Water section).
Other routes of exposure such as inhalation (industrial sites manufac-
turing the esters), dermal exposure, consumption of certain fatty or fatty-
like foods and certain fish will be the major contributors to the body-load
of phthalate esters. Phthalate ester residues in foods such as margarine,
cheese and milk may, on some occasions, reach 50 ppm. Also a special group
at risk will be patients to whom chronic transfusions of blood and blood
products are administered.
Although it is recognized that routes of exposure other than water con-
tribute more to the body burden of phthalate esters, this information will
not be considered in forming ambient water quality criteria until additional
analysis can be made. Therefore, the criteria presented assumed a risk
estimate based only on ambient water exposure.
The need for more accurate determination of residue content of foods,
fish, and water is still very apparent and, as more data become available, a
reevaluation should be made as to the possible hazard to the population by
the ingestion of phthalate esters.
In summary, based on the use of chronic toxicologic data and uncertainty
factors of 100, the criteria levels for phthalate esters have been estab-
lished. The percent contribution of drinking water and of ingesting con-
taminated fish is given in Table 9. Also given are the criteria levels
recommended if exposure is assumed to be from fish and shellfish products
alone.
C-60
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TABLE 9
Summary of Criterion Formulation
o
1
en
i—1
Esters
Dimethyl
Diethyl
Oibutyl
Oi-2-ethylhexyl
Criterion Level
mg/1
313
350
34
15
% Contribution
of Drinking Water
90
81
78
70
% Contribution
of Fish Products
10
19
22
30
Criteria of Exposure
if from Fish Alone
mg/1
2,901
1,842
154
50
-------
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