823R78001
PHTHALATE ESTERS
Ambient Water Quality Criteria
Criteria and Standards Division
Office of Water Planning and Standards
U.S. Environmental Protection Agency
Washington, D.C.
-------
CRITERION DOCUMENT
PHTHALATE ESTERS
CRITERIA
Aquatic Life
For freshwater aquatic life, no criterion for any phthalate
ester can be derived using the Guidelines, and there are insuffi-
cient data to estimate a criterion using other procedures.
For saltwater aquatic life, no criterion for any phthalate
ester can be derived using the Guidelines, and there are insuffi-
cient data to estimate a criterion using other procedures.
Human Health
For the protection of human health from the toxic properties
of phthalate esters ingested through water and through contami-
nated aquatic organisms, the ambient water criteria for dimethyl
phthalate and diethyl phthalate are determined to be 160 mg/1 and
60 mg/1, respectively. The water quality criteria for dibutyl
phthalate and di-2-ethylhexyl phthalate are determined to be 5
mg/1 and 10 mg/1, respectively.
-------
Introduction
Phthalic acid esters or "phthalate esters" represent a
large family of chemicals widely used as plasticizers, pri-
marily in the production of polyvinyl chloride (PVC) resins
(U.S. Int. Trade Comm. 1977). 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 plas-
tics 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 cumulative 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 esti-
mated 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 (DBF) diethyl phthalate (DEP), dimethyl phthalate
(DMP), di-tridecyl phthalate and n-hexyl n-decyl phthalate
(U.S. Int. Trade Comm. 1978).
A-l
-------
PVC resins are used in such diverse industries as con-
struction (high temperature electrical wire, cable insula-
tion, and flooring), home furnishings (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 car-
riers, cosmetics, fragrances, munitions, industrial oils, and
insect repellants (U.S. Int. Trade Comm. 1978).
PAE plasticizers can be present in concentrations up to
60 percent of the total weight of the plastic. The plasti-
cizers are loosely linked to the plastic polymers and are
easily extracted (Mathur, 1974).
For the most part, the esters are colorless liquids,
have low volatility, and are poorly soluble in water but
soluble in organic solvents and oils.
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.
A-2
-------
The ease of extraction of phthalate esters and their
widespread use in PVC or alone account for their ubiquity.
PAEs have been detected in soil (Ogner and Schnitzer, 1970),
water (Ewing and Chian, 1977; Corcoran, 1973; Kites 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 vegetation, animals and minerals (Mathur, 1974;
Graham, 1973), and in areas remote from industrial sites
(Carpenter and Smith, 1972) have raised questions about pos-
sible natural origins of PAEs. PAEs found in greatest fre-
quencies 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 survey were diethyl phthal-
ate, disobutyl phthalate, and diocyl phthalate.
PAEs have been reported to be acutely and chronically
toxic to freshwater and marine aquatic organisms (U.S. EPA,
1978; Mayer and Sanders, 1973). Levels of PAE residues de-
tected in fish 'from ambient waters have not been correlated
with adverse biological effects (Johnson, et al. 1974). Data
show that phthalate esters can be chronically toxic to aqua-
tic organisms at low concentrations. DEHP impairs reproduc-
tion in Daphnia magna by 60 percent at a concentration as low
as 3 uq/1 (Mayer and Sanders, 1973). Toxicological investi-
gations in mammals show that phthalates have low acute toxi-
cities but induce serious chronic effects including terato-
genicity and mutagenicity (Peakall, 1975).
A-3
-------
Due to their large production volumes, ubiquity, and
toxicity to aquatic organisms and mammals, PAE levels in
water should be controlled to prevent potential hazards to
man and aquatic life.
A-4
-------
REFERENCES
Carpenter, E., and K. Smith. 1972. Plastics on the Sargasso
sea surface. Science 175: 1240.
Cocoran, E. 1973. Gas chromatographic detection of phthalate
acid esters. Environ. Health Perspect. 3: 13.
Ewing, B., and E. Chian. 1977. Monitoring to detect pre-
viously 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.
Kites, 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.
Johnson, B., et al. 1974. Dynamics of phthalic acid esters
in aquatic organisms. Page 283. In I.E. Suffet, ed., Fate of
pollutants in air and water environments. Part 2. Wiley
Interscience Publishers, New York.
A-5
-------
Mathur, S. 1974. Phthalate esters in the environment: Pol-
lutants or natural products? Jour. Environ. Quality 3: 189.
Mayer, F.L. 1976. Residue dynamics of di-2-ethylhexylphthal-
ate in fathead minnows, Pimephales promelas. Jour. Fish.
Res. Board Can. 33: 2610.
Mayer, F.L. Jr., and H.O. Sanders. 1973. Toxicology of
phthalic acid esters in aquatic organisms. Environ. Health
Perspect. 3: 153.
Nazir, D., et al. 1971. Isolation, identification, and spe-
cific localization of di-2-ethylhexyl phthalate in bovine
heart muscle mitochondria. Biochemistry 10: 4425.
Ogner, G., and M. Schnitzer. 1970. Humic substances: Fulvic
acid - dialkyl phthalate complexes and their role in pollu-
tion., Science 170: 317.
Peakall, D. 1975. Phthalate esters: Occurence 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 plate-
lets 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
A-6
-------
U.S. EPA. 1978. In-depth studies on health and environmental
impacts of selected water pollutants. Contract No. 68-01-
4646.
U.S. International Trade Commission. 1978. Synthetic organic
chemicals, U.S. production and sales. Washington, D.C.
A-7
-------
AQUATIC LIFE TOXICOLOGY*
FRESHWATER ORGANISMS
Introduction
A limited number of applicable reports were found having
data fcr the effects of phthalate esters on freshwater aquatic
life. More information is available for di-n-butyl and di-2-
ethylhexyl phthalate than for other esters.
Acute Toxicity
All acute values were determined with static procedures and
the test concentrations were unmeasured. Data for five phthalate
esters are in Tables 1 and 2. Values for four of the esters were
from tests with both fish and invertebrate species.
The Final Invertebrate Acute Value of 450 ug/1 for di-2-
ethylhexyl phthalate is derived from a test with Daphnia magna.
Additional acute data for this ester are in Table 6, but the LC50
values for the bluegill and scud exceeded the highest concentra-
tions tested.
*The reader is referred to the Guidelines for Deriving Water
Quality Criteria for the Protection of Aquatic Life [43 FR 21506
(May 18, 1978) and 43 FR 29028 (July 5, 1978)] 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 each table are the calcula-
tions for deriving various measures of toxicity as described in
the Guidelines.
B-l
-------
Tests with butylbenzyl, diethyl and dimethyl phthalates were
conducted with both bluegill and Daphnia magna by the U.S. EPA
(1978). For both species the adjusted LC50 values are within one
order of magnitude and range from 23,672 to 78,200 ug/1' The
Final Acute Values were from the invertebrate tests and are 3,700,
2,100, and 1,300 ug/1, respectively.
Acute di-n-butyl phthalate tests were conducted with four
fish species. The adjusted LC50 values vary from 399 to 3,537
ug/1 or by about nine times. Bluegills were the most sensitive
fish species tested with this ester. The Final Acute Value was 85
ug/1 and derived from the scud invertebrate test since it was the
lowest obtained value. An additional acute datum for this ester
is in Table 6, but the LC50 value exceeded the highest test con-
centration.
Chronic Toxicity
A di-2-ethylhexyl phthalate embryo-larval test was conducted
with rainbow trout (Table 3). The lowest adverse effect concen-
tration from this flow-through test was 14 ug/l« The Final Fish
Chronic Value (0.63 ug/1) is obtained by dividing the chronic
value (4.2 ug/1) by the sensitivity factor (6.7).
Mayer and Sanders (1973) conducted a chronic test with di-2-
ethylhexyl phthalate and Daphnia magna. Significant reproductive
impairment was found at 3 ug/1 (Table 4). Since this value was at
the lowest test concentration, the adverse effects on reproduction
were less than 3 ug/1. After this concentration is divided by the
species sensitivity factor (5.1) a Final Invertebrate Chronic
Value of less than 0.59 ug/1 is obtained. This concentration is
B-2 ./
-------
lower than the comparable value for fish or any plant effects and,
since there is no Residue Limited Toxicant Concentration, the
Final Chronic Value for di-2-ethylhexyl phthalate is less than
0.59 ug/l<>
Plant Effects
The adverse effects of three phthalate esters on the alga,
Selenastrum capricornutumy have been determined (Table 5). Simi-
lar EC50 values were found for cell numbers and chlorophyll a_ for
each ester tested. The lowest EC50 values for diethyl and di-
methyl phthalate were 85,600 and 39,800 v.g/1, respectively. A
much lower EC50 value of 110 ug/1 was obtained with butylbenzyl
phthalate. By comparison^ the adjusted LC50 values found in
Tables 1 and 2 for all three of these esters were within a factor
of 4.
Residues ,
Bioconcentration factors for five phthalate esters have been
reported (Table 6). Mayer (1976) measured both the actual concen-
trations and 14C-labeled di-2-ethylhexyl phthalate in a test
system and found that the difference was less than two times after
equilibrium in fathead minnows,,
Bioconcentration factors for 14C-labeled butylbenzyl-
phthalate, diethyl phthalate, and dimethyl phthalate and bluegills
were 663, 117, and 57, respectively after a 21-day exposure (U.S.
EPA, 1978)„ The half-life of these three phthalate esters was
between 1 and 2 days-
Bicaccumulation data with di-n-octyl phthalates by Sanborn,
et al. (1975) in a static model ecosystem are found in Table 7.
B-3
-------
Their water concentrations rapidly decreased with time and do
not permit comparisons with values in Table 6.
Since no maximum permissible tissue levels exist for phthal-
ate esters, no Residue Limited Toxicant Concentration could be
calculated for any phthalate ester.
Miscellaneous
Additional toxicity data for phthalate esters can be found in
Table 6. Many of these data have been discussed and do not alter
the final acute or chronic values. Mayer, et al. (1977) exposed
rainbow trout eggs to di-2-ethylhexyl phthalate for 90 days and
found concentrations of 14 and 54 ug/1 significantly increased
total protein catabolism 24 days after hatching. This concentra-
tion range is similar to the lowest adverse test concentration
found with this ester in the embryo-larval test (Table 3).
B-4
-------
CRITERION FORMULATION
Freshwater-Aquatic Life
gummary of Available Da.ta
All concentrations below have been rounded to two significant
figures„
butylbenzyl phtha3.ate
Final Fish Acute Vaj.ue = 5,100 ug/1
Fir.:.'. Invertebrate Acute Value = 3,700 ug/1
Final Acute Value = 3,700 ug/1
Final Fish Chronic \ r>juie = not available
Final Invertebrate Chronic Value - not available
Final Plant Value = 110 ug/1
Re3idua Limited Toxicant Concentration = not available
Final Chronic Value - 110 ug/1
0<,44 x Final Acute Value = 1,600 ug/1
dj.ethyl phthalate
Final Fish Acute Value = 14,000 ug/1
final Invertebrate Acute Value = 2,100 ug/1
Final Acute Value = 2,100 ug/1
Final Fish Chronic Value = not available
Final Invertebrate Chronic Value = not available
Final Plant Value = 86^000 ug/1
Residue Limited Toxicant Concentration = not available
Final Chronic Value = 86,000 ug/1
Oo44 x Final Acute Value = 920 ug/1
dimethyl phthalate
Final Fish Acute Value = 6,900 ug/1
Final Invertebrate Acute Value = 1,300 ug/1
-------
Final Acute Value = 1,300 ug/1
Final Fish Chronic Value = not available
Final Invertebrate Chronic Value = not available
Final Plant Value = 39,000 ug/1
Residue Limited Toxicant Concentration = not available
Final Chronic Value = 39,000 ug/1
0.44 x Final Acute Value = 570 ug/1
di-n-butyl phthalate
Final Fish Acute Value = 310 ug/1
Final Invertebrate Acute Value = 36 y.g/1
Final Acute Value = 36 ug/1
Final Fish Chronic Value = not available
Final Invertebrate Chronic Value = not available
Final Plant Value = not available
Residue Limited Toxicant Concentration = not available
Final Chronic Value = not available
0.44 x Final Acute Value = 16 ug/1
di-2-ethylhexylphthalate
Final Fish Acute Value = not available
Final Invertebrate Acute Value = 450 ug/1
Final Acute Value = 450 ug/1
Final Fish Chronic Value =0.63 ug/1
Final Invertebrate Chronic Value = less than 0.59 ug/1
Final Plant Value = not available
Residue Limited Toxicant Concentration = not available
Final Chronic Value = less than 0.59 ug/1
0.44 x Final Acute Value = 200 ug/1
B-6
-------
No freshwater criterion can be derived for any phthalate
ester using the Guidelines because no Final Chronic Value for
either fish or invertebrate species or a good substitute for
either value is available, and there are insufficient data to
estimate a criterion using other procedures.
B-7
-------
Table 1. Freshwater fish acute values for phthalate esters
DO
I
00
Organism
Bluegill.
Lfcpomis macrochirus
Bluegill,
Lepomis macrochirus
Bluegill,
Lepomis macrochirus
Rainbow trout,
Salmo gairdneri
Fathead minnow,
Pimephales promelas
Channel catfish,
Ictalurus punctatus
Bluegill,
Lepomis macrochirus
Bluegill,
Lepomis macrochirus
Bioaseay Test Time LC50
Method" Cone.** (hrs^ (uq/11
Butylber.zyl phthalate
S U 96 43,300
Diethyl phthalate
S U 96 98,200
Dimethyl phthalate
S U 96 49,500
S
S
S
S
S
di-n-butyl phthalate
U 96 6.470
U
U
U
U
96
96
96
96
1.300
2.910
730
1.200
Adjusted
LC50
(uq/11 Keference
23,672 U.S. EPA. 1978
53.686 U.S. EPA. 1978
27,062 U.S. EPA, 1978
3,537 Mayer & Sanders, 1973
711 Mayer fi. Sanders, 1973
1,591 Mayer & Sanders, 1973
399 Mayer & Sanders, 1973
656 U.S. EPA, 1978
* S » static
** U = unmeasured
Geometric mean of adjusted values:
butylbenzyl phthalate = 23,672 yg/1
diethyl phthalate = 53,686 pg/1
dimethyl phthalate = 27.062 ug/1
di-n-butyl phthalate = 1,196 ,ig/l
= 6,100 wg/1
14,000 i'g/1
6,900 pg/1
310 Mg/1
-------
Table 2. Freshwater invertebrate acute values for phthalate esters
DO
I
Cladoceran,
Daphnia maena
Cladoceran,
Daphnia maena
Cladoceran,
Daphnia magna
Cladoceran,
Daphnia maena
Scud,
Gammarus pseudolimnaeus
biocissay Test Time LC50
MetliQU* C O HC ." * (HIS) (Uq/J
Butylbenzyl phthalate
S U 48 92.300
Diethyl phthalate
S U 48 52,100
Dimethyl phthalate
S U 48 33,000
di-2-ethylhexylphthalate
S U 48 11,100
di-n-butyl phthalate
S U 48 2.100
Adjusted
LCbU
(uq/i) Ketfcience
78.200 U.S. EPA, 1978
44,100 U.S. EPA, 1978
28,000 U.S. EPA, 1978
9,400 U.S. EPA, 1978
765 Mayer & Sanders, 1973
* S = static
** U = unmeasured
Geometric mean of adjusted values:
butylbenzyl phthalate = 78,200 \ig/l 78A200 = 3,700 Pg/1
diethyl phthalate = 44,100 wg/1 ^^M = 2,100 pg/1
dimethyl phthalate = 28,000 yg/1 28P° = 1,300 pg/1
di-2-ethylhexylphthalate = 9,400 pg/1
= 450 yg/1
~
di-n-butyl pththalate = 765 Mg/1 ~2T = 36 ng/1
-------
Table 3. Freshwater fish chronic values for phthalate esters (Mehrle & Mayer, 1976)
Chronic
Limits Value
Organism Test* (ng/i) (tui/i)
di-2-ethylhexylphthalate
Rainbow trout. E-L 5-14 4.2
Salmo gairdneri
* E-L = embryo-larval
4.2
Geometric mean of chronic values = 4.2 yg/1 7—•= = 0.63|iJg/l
Lowest chronic value = 4.2 yg/1
CD
I
-------
Table 4. Freshwater invercebrace chronic values for phthalate esters (Mayer & Sanders, 1973)
Organism
Cladoceran,
Daphnia magna
Chronic
Limits Value
Test* (uq/i) (uq/i)
di-2-ethylhexylphthalate
LC <3.0
<3.0
* LC = life cycle
Geometric mean of chronic values = <3.0 yg/1
Lowest chronic value = <3.0 yg/1
<3.0
*0.59 pg/1
DO
I
-------
Table 5. Freshwater plant effects for phthalate esters (U.S. EPA. 1978)
Organism
Effect
Concentration
(UR/I)
Butylbenzyl phthalate
DO
\->
NJ
Alga,
Selenastrum
capricornutum
Alga,
Selenastrum
capricornutum
Alga,
Selenastrum
capricornutum
Alga,
Selenastrum
capricornutum
Alga,
Selenastrum
capricornutum
Alga,
Selenastrum
capricornutum
EC50 96-hr
chlorophyll a
EC50 96-hr
cell number
110
130
Diethyl phthalate
90,300
EC50 96-hr
chlorophyll a
EC50 96-hr
cell number
85,600
Dimethyl phthalate
42,700
EC50 96-hr
chlorophyll a
EC50 96-hr
cell number
39,800
Lowest plant value: butylbenzyl phthalate = 110 pg/1
diethyl phthalate = 85,600 ng/1
dimethyl phthalate = 39,800 pg/1
-------
Table 6. Freshwater residues for phthalate esters
00
M
U>
Organism
Cladoceran,
Daphnla magna
Scud,
Gammarus pseudolimnaeus
Scud,
Gammarus pseudolimnaeus
Sowbug ,
Asellus brevicaudus
Rainbow trout,
Salmo gairdneri
Fathead minnow,
Pimephales promelas
Fathead minnow,
Pimephales promelas
Bluegill,
Lepomis macrochirus
Bluegill,
Lepomis macrochirus
Bluegill,
Lepomis macrochirus
Bioconcentration Factoi*
di-n-butyl phthalate
400
1,400
di-2-ethylhexylphthalate
54-2,680--*
14-50**
42-113
155-886
91-569***
butylbenzylphthalate
663
diethylphthalate
117
dimethylphthalate
57
Time
(days)
14
14
14-21
21
36
56
56
21
21
21
neterence
Mayer & Sanders
Mayer & Sanders
Sanders, et al .
Sanders, et al.
Mehrle & Mayer,
Mayer, 1976
Mayer, 1976
U.S. EPA. 1978
U.S. EPA, 1978
U.S. EPA, 1978
, 1973
, 1973
1973
1973
1976
Based on total l>tC radioactivity accumulated.
"*" Conversion from dry to wet weight.
***Based on measured concentrations of di-2-ethylhexylphthalate.
-------
Organism
Table 7. Other freshwater data'for phthalate esters
Test
Duration Ettect
Result
(ug/1) Retereiicfe
di-2-ethylhexylphthalate
Scud, 96 hrs LC50
Gammarus pseudolimnaeus
>32.000 Sanders, et al. 1973
DO
I
Rainbow trout,
Saltno gairdneri
Guppy,
Poecilla reticulatus
Bluegill,
Lepomis macrochlrus
Crayfish,
Orconectes nais
Alga,
Oedogonium cardiacum
Cladoceran,
Daphnia maena
Mosquito (larva),
Culex pipeus
quinquefasciatus
Snail,
Physa sp
Mosquitofish,
Gambusia affinis
24 days Significant increase 14-54
in total body protein
catabolism
90 days Incre'ase in
aborted young
96 hrs LC50
di-n-butyl phthalate
96 hrs LC50
di-n-octyl phthalate
33 days Model ecosystem*
28.500X
bioconcentration
33 days Model ecosystem*
2.600X
bioconcentration
33 days Model ecosystem*
9.400X
bioconcentration
33 days Model ecosystem*
13.600X
bioconcentration
33 days Model ecosystem*
9.400X
bioconcentration
Mayer, et al. 1977
fed 100 Mayer & Sanders, 1973
ug/g in
diet
>770.000 U.S. EPA, 1978
>10,000 Mayer & Sanders. 1973
Sanborn, et al. 1975
Sanborn, et al. 1975
Sanborn, et al. 1975
Sanborn, et al. 1975
Sanborn, et al. 1975
* Based on actual concentrations of di-n-octyl-phthalate accumulated
-------
SALTWATER ORGANISMS
Introduction
Phthalate esters have contaminated various segments of our en-
vironment, including aquatic organisms and water (Mayer and Sanders,
1973), and there is a growing concern that they may be a menace to
health and to our ecological system. Phthalate esters are a large
group of chemical agents (esters of ortho benzene dicarboxylic acid)
used primarily as plasticizers.
Toxicity test data for saltwater organisms are available for
only four phthalate esters. Laughlin, et al. (1977) conducted studies
on the effects of di-n-butyl phthalate and dimethyl phthalate on
development of the mud crab, Rhithropanopeus harrisii. All other data
(U.S. EPA, 1978) consist of LC50 or EC50 values based on static tests
and unmeasured concentrations for three species (sheepshead minnow,
Cyprinodon variegatus; mysid shrimp, Mysidopsis bahia; and an alga,
Skeletonema costatum) for butylbenzyl phthalate, diethyl phtha- late,
and dimethyl phthalate. These data indicate great differences in
toxicity among esters; therefore, it would be inappropriate to gener-
ate a criterion for phthalate esters as a group.
Acute Toxicity
Butylbenzyl phthalate and diethyl phthalate were less toxic to
the sheepshead minnow than they were to the mysid shrimp; dimethyl
phthalate was more toxic (Tables 8 and 9). Unadjusted 96-hour LC50
values for butylbenzyl, diethyl, and dimethyl phthalates for the
sheepshead minnow were 445,000, 29,600, and 58,000 ug/1, respectively,
B-15
-------
and for the mysid shrimp, 9,630, 7,590, and 73,700 ug/1/ respectively
(U.S. EPA, 1978).
When the geometric means of the adjusted LC50 values for fishes
are divided by the species sensitivity factor (3.7), the resultant
Final Fish Acute Values are 66,000 ug/1 for butylbenzyl phthalate,
4,400 ug/1 for diethyl phthalate, and 8,600 ug/1 for dimethyl phthalate
(Table 8). The geometric means of the adjusted LC50 values for inver-
tebrate species, when divided by the species sensitivity factor (49),
give Final Invertebrate Acute Values of 170 ug/1 for butylbenzyl
phthalate, 130 ug/1 for diethyl phthalate, and 1,300 ug/1 for dimethyl
phthalate. Freshwater acute toxicity data (Tables 1 and 2) for butyl-
benzyl, diethyl, and dimethyl phthalates showed that toxicity to fresh-
water fish and invertebrate species did not differ greatly from that to
saltwater animals, although relative sensitivity of freshwater fish and
invertebrate species to phthalate esters usually differed from salt-
water organisms.
Chronic Toxicity
No saltwater fish or invertebrate species has been tested in a
chronic toxicity study.
Plant Effects
Butylbenzyl phthalate and dimethyl phthalate were more toxic to a
saltwater alga, Skeletonema costatum, than to the tested fish and
invertebrate species; diethyl phthalate was less toxic (Table 10).
Butylbenzyl phthalate was particularly toxic to the alga: a concen-
tration of 170 ug/1 caused 50 percent reduction in chlorophyll a_ and
190 ug/1 caused 50 percent reduction in cell numbers in 96 hours (U.S.
EPA, 1978). Freshwater data (Table 5) also indicated that a fresh-
water alga was especially sensitive to butylbenzyl phthalate. The
B-16
-------
96-hour EC50 values for Skeletonema costatum exposed to dimethyl
phthalate were 26,100 ug/1 for chlorophyll a and 29,800 ug/i for cell
numbers. Exposure to diethyl phthalate resulted in a 96-hour EC50 of
65,500 ug/1 for chlorophyll a and a 96-hour EC50 of 85,000 ug/1 for
cell numbers (U.S. EPA, 1978).
Residues
No data for bioconcentration of phthalate esters by saltwater
species are available.
Miscellaneous
In laboratory experiments by Laughiin, et al. (1977), 1,000 ug/1
di-n-butyl phthalate or dimethyl phthalate had no significant effect on
the entire larval development of the mud crab, Rhithropanopeus harrisii
(Table 11). There are no other saltwater data that suggest more sen-
sitive effects than those already presented.
B-17
-------
CRITERION FORMULATION
Saltwater-Aquatic Life
Summary of Available Data
The concentrations below have been rounded to two significant
figures.
butylbenzyl phthalate
Final Fish Acute Value = 66,000 ug/1
Final Invertebrate Acute Value = 170 ug/1
Final Acute Value = 170 ug/1
Final Fish Chronic Value = not available
Final Invertebrate Chronic Value = not available
Final Plant Value = 170 ug/1
Residue Limited Toxicant Concentration = not available
Final Chronic Value = 170 ug/1
•.
0.44 x Final Acute Value = 75 ug/1
diethyl phthalate
Final Fish Acute Value = 4,400 ug/1
Final Invertebrate Acute Value = 130 ug/1
Final Acute Value = 130 ug/1
Final Fish Chronic Value = not available
Final Invertebrate Chronic Value = not available
Final Plant Value = 66,000 ug/1
Residue Limited Toxicant Concentration = not available
Final Chronic Value = 66,000 ug/1
0.44 x Final Acute Value = 57 ug/1
dimethyl phthalate
Final Fish Acute Value = 8,600 ug/1
Final Invertebrate Acute Value = 1,300 ug/1
B-18
-------
Final Acute Value = 1,300 ug/1
Final Fish Chronic Value = not available
Final Invertebrate Chronic Value = not available
Final Plant Value = 26,000 ug/1
Residue Limited Toxicant Concentration = not available
Final Chronic Value = 26,000 ug/1
0.44 x Final Acute Value - 570 ug/1
No saltwater criterion can be derived for any phthalate ester
using the Guidelines because no Final Chronic Value for either fish
or invertebrate species or a good substitute for either value is
available, and there are insufficient data to estimate a criterion
using other procedures.
\B-19
-------
Table 8. Marine fish acute values for phthalate esters (U.S. EPA, 1978)
ro
to
o
Organism
Sheepshead minnow
(j uvenilc),
Cyprinodon variegatus
Sheepshead minnow
(juvenile),
Cyprinodon variegatus
Sheepshead minnow
(juvenile),
Cyprinodon variegatus
Adjusted
Bioaseay Test Time LC50 LC50
Method * Cone. ** (hrs) (ug/1) (ug/i)
Butylbenzyl phthalate
S U 96 445.000 243,282
Diethyl phthalate
U 96 29.600 16.182
Dimethyl phthalate
U 96 58.000 31,709
* 'S = static
** U = unmeasured
Geometric mean of adjusted values: butylbenzyl phthalate «
diethyl phthalate - —a
dimethyl phthalate =
243.282
—3-7— = 66,000 tJg/1
j— = 4,400 Mg/1
U^. = 8,600 pg/1
-------
Table 9. Marine invertebrate acute values for phthalate esters (U.S. EPA, 1978)
Organism
Mysid shrimp,
Mysidopsis bahia
Mysid shrimp,
Mysidopsis bahia
Mysid shrimp,
Mysidopsis bahia
bioassay Test Time
Method * Cone.** if'is)
LC50
(uq/J.)
U
Piethyl phthalate
96 7,590-
Adjusted
LCbO
Butylbenzyl phthalate
U 96 9.630 8,157
6,429
Dimethyl phthalate
U 96 73,700 62,424
DO
tsj
* S = static
**U = unmeasured.
ft I 5 7
Geometric mean of adjusted values: butylbenzyl phthalate = '^ = 170 yg/1
diethyl phthalate - Ao ~ 13° "8/1
dimethyl phthalate - 64^>2A = 1,300 pg/1
-------
Table 10- Marine plane effects for phthalate esters (U.S. EPA, 1978)
03
I
to
NJ
Organism
Alga.
Skeleconema costatura
Alga,
Skeletonema costatum
Alga,
Skeletonema costatum
Alga,
Skeletonema costatum
Alga,
Skeletonema costatum
Alga,
Skeletonema costatum
Etfect
chlorophyll a
EC50 after 96
hr
Cell numbers
EC50 after 96
hr
Chlorophyll a
EC50 after 96~
hr
Cell numbers
EC50 after 96
hr
Chlorophyll a
EC50 after 96
hr
Cell numbers
EC50 after 96
hr
Concentration
(uq/i)
Butylbenzyl phthalate
170
190
Diethyl phthalate
65,500
85.000
Dimethyl phthalate
26.100
29.800
Lowest plant value = butylbenzyl phthalate = 170 Mg/1
diethyl phthalate = 65,500 ug/1
dimethyl phthalate = 26.100 gg/1
-------
to
I
Table 11, Other marine data for phthalate esters (Laughlin. et al. 1977)
Organism
Test
Duration Effect
Result
(uq/11
Reference
Mud crab (larva),
Rhithropanopeus
Harrisii
Entire
larval
development
Di-n-butyl phthalate
None on development 1,000
Laughlin, et al. 1977
Mud crab (larva),
Rhithropanopeus
harrissii
Entire
larval
development
Dimethyl phthalate
None on development
1,000 Laughlin, et al. 1977
-------
PHTHALATES
REFERENCES
Laughlin, R.B., et al. 1977. Effects of polychlorinated
biphenyls, polychlorinated napthalenes, and phthalate esters
on larval development of the mud crab Rhithropanopeus harrisii.
Pages 95-110. Ir\ Pollutant effects on marine organisms.
D.C. Health Co., Lexington, Mass.
Mayer, F.L. 1976. Residue dynamics of di-2-ethylhexylphtha-
late in fathead minnows (Pimephales promelas). Jour. Fish.
Res. Board Can. 33: 2610.
Mayer, F.L. Jr., and H.O. Sanders. 1973. Toxicology of
phthalic acid esters in aquatic organisms. Environ. Health
Perspect. 3: 153.
Mayer, F.L., et al. 1977. Collagen metabolism in fish
exposed to organic chemicals. Pages 31-54. In Recent advances
in fish toxicology, a symposium. EPA 600/3-77-085. U.S.
Environ. Prot. Agency, Corvallis, Oe.
Mehrle, P.M., and F.L. Mayer. 1976. Di-2-ethylhexylphthalate:
Residue dynamics and biological effects in rainbow trout
and fathead minnows. Pages 519-524. Ir\ Trace substances in
environmental health. University of Missouri Press, Columbia.
B-24
-------
Sanborn, 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 al. 1973. Toxicity, residue dynamics,
and reproductive effects of phthalate esters in aquatic
invertebrates. Environ. Res. 6: 84.
U.S. EPA. 1978. In-depth studies on health and enviromental
impacts of selected water pollutants. Contract No. 68-01-
4646.
B-25
-------
Mammalian Toxicology and Human Health Effects
EXPOSURE
Introduction
The annual production of phthalic acid esters in the
United States in 1977 amounted to approximately 1.2 billion
pounds. Table 1 lists the major esters with their production
figures. Since 1945, the cumulative 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 iso-
meric forms of benzenedicarboxylic acid esters are also pro-
duced. 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" 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 contaminates 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.
-------
TABLE 1
Production of Individual Phthalic Acid Esters
in U.S. in 1977
Ester Production in Pounds
(1000 pounds)
Dibutyl 16,592
Diethyl 17,471
Diisodecyl 160,567
Dimethyl 9,887
Dioctyl
Di-2-ethylhexyl 388,543
Other dioctyl phthalates 11,664
Di-tridecyl 23,278
n-Hexyl n-decyl 15,182
All other phthalate esters 559,229
Total 1,202,413
From:United States International Trade Commission,
U.S. Government Printing Office, Washington, 1978,
USITC Publication 920, p. 263.
Pthalic acid esters have a large number of commercial
uses, the largest being as plasticizers for specific plas-
tics such as polyvinyl chloride. Other uses for these esters
include: defoaming agents in the production of paper, in cos-
metic products as a vehicle (primarily diethyl phthalate) for
perfumes, in lubricating oils, and in other industrial and
consumer applications. Table 2 illustrates the variety of
uses for esters with an estimate of the amount of the esters
c-2
-------
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 Hf7
Home Furnishings
Furniture upholstery 90
Wall coverings 38
Houseware . 30
Miscellaneous 45
Subtotal 203
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
From:Graham,1973.
C-3
-------
used in the specific categories. Approximately 20 different
esters are used in the specific categories.
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 (DOP) and di-2-
ethylhexyl phthalate (DEHP) are often used synonymously even
though it should be clear that they are not the same, one
being the isomer of the other.
For the most part, the esters are colorless liquids,
have low volatility, and are poorly soluble in water but
soluble in organic solvents and oils. Table 3 lists several
of the physical properties of these esters.
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 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) through paren-
teral administration (via blood bags and tubes in which the
ester is extracted by a parenteral solution including blood).
C-4
-------
TABLE 3
Physical and Chemical Properties of Phthalate Esters
Compound Molecular
.Weight
Dimethyl
phthalate
Diethyl
phthalate
Diallyl
phthalate
Diisobutyl
phthalate
Dibutyl
phthalate
Dimethoxy ethyl
phthalate
Dicyclohexyl
phthalate
Butyl octyl
phthalate
Dihexyl
phthalate
Butj-lphthalyl
butyl glycolate
Dibutoxyethyl
ethyl phthalate
Di-2-ethylhexyl
phthalate
Diisooctyl
phthalate
Di-r.-octyl
phthalate
Dincnyl
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 Bp,
Gravity °C
1.189 (25/25) 282
1.123 (25/4) 296.1
1.120 (20/20) 290
1.040 327
1.0465 (21) 340
1.171 (20) 190-210
1.20 (25/25) 220-228
340
0.990
1.097 (25/25) 219/5 nun/
1.063 210
0.985 (20/20) 386.9/5 mm
0.981 239/5 mm
0.978 220/5 mm/
0.965 413
Solubility in
H20, g/100 ml
0.5
Insoluble
0.01
Insoluble
0.45 (25°C)
0.85
Insoluble
—
Insoluble
0.012%
0.03
Insoluble
Insoluble
Insoluble
Insoluble
C-5
U •
-------
Ingestion from Water
In the early seventies, a great deal of attention began
to focus on chemical contaminants in surface water and adja-
cent ocean regions. One of the first reports published on the
presence of phthalic acid esters was presented 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 calculated 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
concentration. Corcoran also indicated the presence of DEHP
(or its equivalent 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 ap-
proximately seven miles from the mouth of the Charles River
the level of phthalate was 1.8 to 1.9 ppb. As the water ap-
proached 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.
C-6
-------
Ingestion from Food
Since a number of packaging materials and tubings used in
the production of foods and beverages are polyvinyl chloride
containing phthalic acid esters, primarily DEHP, the esters
can migrate to the food and beverages. The extent of migra-
tion 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. Extrac-
tion 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 DEHP 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 approximately 40 mg of DEHP could be extracted
over a 15-day period from tubings in contact with milk in
actual practice but went on to indicate 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 dicyclo-
hexyl phthalates) extracted less than one percent of the
esters after one month at 25°C. The concentrations in the
food were reported as less than 2 ppm.
Food and Drug Administration surveys indicate that sev-
eral of the phthalate esters are present in 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 4,
C-7
-------
also been reported. Table 4, taken from the study by Tomita,
et al. (1977) shows the amounts of several agents migrating to
selected Japanese foods packaged in plastics, laminated films,
paper, and aluminum foil. As will be noted, levels above 600
ppm and even higher than 3000 ppm of total phthalates migrated
to certain foods.
A bioconcentration factor (BCF) relates the concentration
of a chemical in water to the concentration in aquatic organ-
isms, but BCF's are not available for the edible portions of
all four major groups of aquatic organisms consumed in the
United States. Since data indicate that the BCF for lipid-
soluble compounds is proportional to percent lipids, BCF's can
be adjusted to edible portions using data on percent lipids
and the amounts of various species consumed by Americans. A
recent survey on fish and shellfish consumption in the United
States (Cordle, et al. 1978) found that the per capita con-
sumption is 18.7 g/day. From the data on the nineteen major
species identified in the survey and data on the fat content
of the edible portion of these species (Sidwell, et al. 1974),
the relative consumption of the four major groups and the
weighted average percent lipids for each group can be calcu-
lated:
Consumption Weighted Average
Group (Percent) Percent Lipids
Freshwater fishes 12 4.8
Saltwater fishes 61 2.3
Saltwater molluscs 9 1.2
Saltwater decapods 18 1.2
C-8
-------
Using the percentages for consumption and lipids for each of
these groups, the weighted average percent lipids is 2.3 for
consumed fish and shellfish.
Measured steady-state bioconcentration factors of 57,
117, and 663 were obtained for dimethyl, diethyl, and butyl-
benzyl phthalates using bluegills containing about one percent
lipids (U.S. EPA, 1978). An adjustment factor of 2.3/1.0 =
2.3 can be used to adjust the measured BCF from the 1.0 per-
cent lipids of the bluegill to the 2.3 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 aquatic organisms consumed by Americans are calculated to
be 130, 270, and 1,500, respectively.
No measured steady-state bioconcentration factor (BCF) is
available for dibutyl phthalate, but the equation "Log BCF =
0.76 Log P - 0.23" can be used (Vieth, et al. Manuscript) to
estimate the BCF for aquatic organisms that contain about
eight percent lipids from the octanol-water partition coeffi-
cient (P). Based on an octanol-water partition coefficent of
760, the steady-state bioconcentration factor for dibutyl
phthalate is estimated to be 91. An adjustment factor of
2.3/8.0 = 0.2875 can be used to adjust the estimated BCF from
the 8.0 percent lipids on which the equation is based to the
2.3 percent lipids that is the weighted average for consumed
fish and shellfish. Thus, the weighted average bioconcentra-
tion factor for dibutyl phthalate and the edible portion of
all aquatic organisms consumed by Americans is calculated to
be 91 x 0.2875 = 26.
C-9
-------
An average measured steady-state bioconcentration factor
of 330 was obtained for di-2-ethylhexyl phthalate using fat-
head minnows containing about eight percent lipids (Mayer,
1976). An adjustment factor of 2.3/8.0 = 0.2875 can be used
to adjust the measured BCF from the 8.0 percent lipids of the
fathead minnow to the 2.3 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 aquatic organisms consumed by
Americans is calculated to be 330 x 0.2875 = 95.
Inhalation
This route may be a significant portal of entrance for
esters of phthalic 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 fur-
nishings, 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 from ship
quarters furnished with PVC tile, decorative laminated plas-
tics and pavinols (assumed to be PVC plastics). He reported
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/m3 at one working
site where mixing was done and a level of 10 to 66 mg/m3 at
C-10
-------
o
TABLE 4
Migration of Phthalic Acid Esters from Packaging Film to Foodstuffs*
Time after
manufacture
Foodstuffs (months)
Tempura (frying) A
powder B
Instant cream A
soup B
C
Instant soybean
soup
Soft margarine
Fried potato A
cake B
C
Orange juice
Red ginger pickles
Sugar
Table salt
1 1 ....... .,.., , — ,
3
4
14
p
4
1
1
?
p
p
Packaging materials (ppm)
Materials**
Pl-L
Pl-L
P-A1-P1
P-Al-Pl
P-A1-P1
P-Pl
Pi
P-PL
P-PL
P-PL
P-Pl
PI
Pi
P-Pl
DNBP
70.28
6.29
23.17
586.16
588.75
2.75
1.29
10.86
10.66
22.98
1.52
3.00
7.24
5.18
DEHP
3675.0
2.30
1.35
58.92
58.93
1.85
1.44
385.85
1.28
11.80
0.74
2.14
2.75
2.58
Total
3745.28
8.59
24.52
647.08
647.08
4.60
2.73
396.91
11.94
34.78
2.26
5.14
9.99
7.76
Foodstuffs (ppm)
DNBP
14.70
0.39
1.73
60.37
51.79
nd
nd
1.11
nd
1.21
0.35
0.11
nd
nd
DEHP
68.08
0.11
0.04
2.15
3.01
nd
nd
0.05
nd
9.06
0.05
nd
nd
nd
Total
82.78
0.50
1.77
62.52
54.80
nd
nd
1.16
nd
10.27
0.40
0.11
nd
nd
From: Tomita, et al. 1977.
** Pi indicates plastic
L indicates laminated film
P indicates paper
Al indicates aluminum foil
-------
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 main-
tained, these workers could inhale sufficient concentrations
of the ester to lead to health problems.
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. Since dimethyl phthal-
ate is used as a mosquito repellent, dermal absorption can
occur. Swimmiig 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 pre-
vents even a very crude projection of the levels of esters
which could enter man through the skin.
Since a number of medical devices such as blood bags, in-
fusion containers, collection and administration tubings, and
C-12
-------
catheters are prepared from plasticized (generally DEHP) poly-
vinyl chloride, a parenteral route of entrance into a selected
human population becomes a possibility. In fact, it is pos-
sible that the parenteral route contributes the greatest quan-
tity 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 ap-
proximately 150 mg of DEHP. In open heart surgery, extra cor-
poreal pump oxygenators are used. Approximately 360,000 such
operations are performed each year. Under these conditions, a
patient may be exposed to an average of 33 mg of DEHP during
the surgery.
As early as 1960, a report appeared by Meyler, et al.
(1960) that certain 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 contained an
organotin stabilizer, the authors surmised that the toxic com-
ponent was the stabilizer and not the phthalate ester.
Braun and Kummel (1963), reported that PVC containers
used for storage of blood and transfusion solutions did re-
lease phthalate esters as well as other additives to an ex-
tracting medium (water).
C-13
-------
A report by Guess, et al. (1967) revealed that a number
of American PVC blood bags containing an anticoagulant solu-
tion (ACD) were contaminated by the presence of small amounts
of DEHP, 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 ai. (1975) identified the presence of DEHP in
neonatal tissues 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 DEHP content of tissue if the speci-
fic patient had also received blood products. Residue levels
were measured in both heart and gastrointestinal tissues. The
average level of DEHP in heart tissue was 1.27 ug/g» In the
gut of the three patients having died of gastrointestinal dis-
orders, 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 solu-
tions 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 lipoidal solu-
tions such as blood and blood fractions are used, the extent
of release becomes significant.
The quantity of di-2-ethylhexyl phthalate released into
C-14
-------
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 5.
Needham and Luzzi (1973) indicated that when PVC infu-
sion 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 pres-
ence of ethyl alcohol in the solution will increase the level
of DEHP in the solution. A ten percent solution will in-
crease 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).
TABLE 5
Extraction Data of DEHP from PVC Containers
1. Normal whole blood stored at 4°C contains 0.19 mg% DEHP
on collection and 5.84 mg% 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%).
3. The level of DEHP in stored platelets maintained at 4°C
22°C after 72 hours is 10.85 mg% and 43.21 mg%, respec-
tively.
Summarized from: Kevy, et al. 1978.
The total quantity of DEHP a transfused patient may re-
ceive parenterally will, of course, depend upon the number of
units of blood or blood products administered to him. Pa-
tients undergoing chronic transfusions with whole blood,
C-15
-------
packed cells, platelets and plasma stored in PVC containers
may receive a total of approximately 70 mg of DEHP. There
are cases, however, 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 intraperi-
toneal cavity, and the lungs. There is also evidence in-
dicating 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
l^c-DEHP 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 intes-
tine. The urine contained about 62 percent in water ex-
tracts. Daniel and Bratt (1974) injected a single oral dose
of 14C-DEHP in rats and found 42 percent and 57 percent
of the dose in the urine and feces, respectively, in seven
days. They also pointed out that a significant amount of the
dose is excreted in bile. In studies by Wallin, 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 radioactivity was
found in the feces and in the gastrointestinal tract contents.
C-16
-------
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 admin-
istered phthalic add esters are absorbed in the gut pri-
marily as monoesters. Wallen, et al. (1974) however, found
from their studies that a significant amount of orally admin-
istered DEHP is absorbed in the gastrointestinal tract as the
intact compound. From the present data, it appears clear
that the diester phthalates can be hydrolyzed to the mono-
ester 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 conditions in several species of animals.
Information on the absorption of the phthalic acid es-
ters in man is limited. As early as 1945, however, Shaffer,
et al. (1945) reported that a single oral dose of 10 g DEHP
in a human subject was recovered as a phthalate equivalent in
the urine after 24 hours. The amount recovered was 4.5 per-
cent of the original dose. In another subject, 5 g of DEHP
was taken orally and 2.0 percent of the original dose (as
phthalate equivalent) found in the urine 24 hours later.
Tomita, et al. (1977) reported the presence of phthalate es-
ters 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 (DNBP)
C-17
^ /
-------
levels detected in the blood after meals were much higher
than prior to eating the foods in the plastic packaging
system. In 13 individuals who were included in the study,
(DEHP and DNBP) in 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 asters of phthalic acid esters (or their metab
olites) distribute quite rapidly to various organs and tis-
sues both in animals and humans. Again, it must be kept in
mind that, depending upon the route and the physical form of
the ester (true solution, colloid, emulsion), the distribu-
tion 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 de-
tected the presence of DEHP in spleen, liver, lung, and ab-
dominal fat with concentrations ranging from 0.025 mg/g in
spleen to 0.270 mg/g in abdominal fat.
Radio-labeled DEHP (emulsified in oleic acid) admin-
istered i.v. as a single dose was found to disappear rapidly
from blood and approximately 60 to 70 percent of the total
dose was detected in the liver and lungs within two hours of
018
-------
administration of the dose (Daniel and Bratt, 1974). In
studies in which rats were maintained on diets containing
DEHP, there was a progressive 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 autoradiography techniques. Results
from the study revealed that a rapid accumulation of radio-
activity in the kidney and the liver had occurred followed by
rapid excretion into urine, bile, and intestine. No accumu-
lation of the compound was found (up to 168 hours after the
injection) in the spleen and lung, but significant radio-
activity was detected in the lumen of the intestine 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 reached a maximum level in the first two
to six hours. Peak blood levels of the compound occurred
about six hours after administration. Intravenous adminis-
tration of labeled DEHP as a dispersion prepared by sonifica-
tion of DEHP in saline led to 70 to 80 percent of the origi-
nal 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
C-19
-------
end of the seventh day. The intestine (after oral and i.v.
administration) revealed a relatively high level of radio-
activity 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 adminis-
tration. 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
DEHP (as radioactivity) in fatty tissue. Application of
labeled diethyl phthalate to the skin of rabbits resulted in
detection of the compound in the lung, heart, liver, kidney,
gonads and spleen after three days. The compound (or its
metabolite) was also detected in the brain but, surprisingly,
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 ap-
parent 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 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
C-20
-------
common to humans revealed the presence of DEHP (or metabo-
lites) in trace amounts even up to 14 months post-transfu-
sion. 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 preventing 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 metabolism is the conversion of the
diester to monoester (mono-2-ethylhexyl phthalate). By {jj-
and (OJ-1) oxidation, the side chain of the monoester forms
two different alcohol intermediates. Further oxidation of
the alcohols leads to the corresponding carboxylic acid or
ketone and, in turn, the acid may be further oxidized (^-oxi-
dation) . Figure 1 shows a number of products which can be
formed from metabolism of orally ingested DEHP (in rats).
Lack of detailed data on the metabolism of other esters in
various species of animals and in humans prevents a clear
understanding of what metabolic products are formed in other
species. It seems clear, however, that for DEHP a signifi-
cant biotransf ormation can take place in the gut (DEHP to the
monoester) and thus the same possibility may also be true in
other higher orders of animals and in man. The absorbed
intact DEHP and/or the monoester is then further metabolized
in the liver.
C-21
-------
o
10
to
DEHP
0
,—C—OH
C—0—CH,—CH—(CH,),—CH,
CH,
CH,
Monocthylhcxyl
phtholntc
O
,— C— OH OH
C—O—CH,—CH—CH,—CH,—C—CHa
CH,^ II
CH,
O
C—0—CH,—CH—CH,—CH,—C—CH,
O CH,
CH,
0
,—C—OH O
C—0—CH,—CH—(CH,),—C—OH
in,,
CH,
0
>C-OH
^^—C— 0— CH,—CH— CH,—C— OH
CH,
CH,
Figure 1. Routes of metabolism of. di(2-ethylhexyl) phthalate (after Albro, et al. 1973)
-------
Excretion
For the most part, the esters of phthalic acid in ani-
mals and man are excreted readily in urine and feces. For
example, Lake, et al. (1975) found that a single oral dose of
labeled DEEP 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 in-
creasing the content of DEHP (or metabolites) in the intes-
tine.
Schulz and Rubin (1973-) administered labeled DEHP i.v.
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, eight
percent of the total injected DEHP was found in water-soluble
metabolites, primarily in the liver, intestinal contents and
urine. Twenty-four hours after injection, 54.6 percent of
the initial dose was recovered as water-soluble metabolites
primarily in the intestinal tract, excreted feces, and urine
and 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).
C-23
-------
They noted that 68 percent and 63 percent, respectively, 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 administra-
tion 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 for 20 days revealed the presence of in-
tact DEHP in the urine. They concluded that "repeated admin-
istration of DEHP may lead to its accumulation in the body
until a steady state is reached between the rates of absorp-
tion and elimination." After steady state is reached, DEHP,
as the unchanged molecule, would appear in the urine.
As Thomas, et al. (1978) have expressed in their review
article on biological effects of DEHP, pharmacokinetic data
in animals and humans support the thesis that DEHP is ab-
sorbed from the gastrointestinal tract and widely distributed
to various tissues following either the oral or i.v. routes
of administration. DEHP 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, Sub-acute, 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
C-24
/ 1
-------
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 Perspectives. As will become
evident, most of the detailed toxicological studies have
centered primarily on DEHP since this specific ester accounts
for approximately 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 ef-
fect of the esters is most likely due to one of the metabo-
lites in particular to the monoester. This appears to be the
case for DEHP since this ester has been studied more exten-
sively than the others. Table 6 is taken from Autian's 1973
review and lists the 1,050 is of the esters. Oral acute tox-
icity for the lower molecular weight esters is greater in
animals than for the higher molecular weight esters such as
DEHP. Other routes of administration such as i.p. and dermal
do not significantly increase the acute toxicity.
The toxicity of DEHP by the i.v. route is quite impor-
tant since, as has been indicated previously, PVC administra-
tion devices will leach the plasticizer into blood and lipo-
protein-containing solutions. Since DEHP has a very limited
solubility in water, other means of administering the agent
C-25
-------
TABLE 6
Acute Toxicity of Phthalate Esters: LD5Q in Animals
(Autian, 1973).
Compound
Dimethyl
phthalate
Diethyl
phthalate
Dime thoxy ethyl
phthalate
Diallyl
phthalate
Dibutyl
phthalate
Diisobutyl
phthalate
Butyl carbobutoxy-
methyl phthalate
Dihexyl
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
Rat
Rabbit
Route
Oral
IP
IP
Oral
IP
Oral
Dermal
IP
IP
IP
Oral
Oral
IP
Oral
IP
Oral
Dermal
IP
Oral
Oral
Dermal
IP
IP
IM
Dermal
Oral
IP
IP
Dermal
Oral
IP
Oral
Dermal
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. Oa
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
30.0
20. Oa
C-26
-------
TABLE 6 (Continued)
Compound
Dioctyl
phthalate
Di-2-ethyhexyl
phthalate
Animal
Mouse
Rat
Guinea pig
Mouse
. Rat
Rat
Rabbit
Guinea pig
Route
Oral
IP
Dermal
IP
Oral
IP
Oral
Dermal
LD50
g/Kg
13.0
50. Oa
5.0a
14.2
26.0
50. Oa
34.0
10.0
Butylbenzyl
phthalate
Dicapryl
phthalate
Mouse
Mouse
IP
IP
3.16
14.2
Dinonyl
phthalate
Dibutyl (diethylene
gylcol bisphthalate)
Dialkyl
phthalate
Rat
Mouse
Mouse
Rat
Rat
Mouse
Rat
Oral
Oral
IP
Oral
IP
Oral
IP
2.00
11.2
11." 2
^11.2
>20.00
>20.00
in ml/kg.
C-27
-------
in experimental animals have been used to study the toxic ef-
fects when administered i.v. Preparation of emulsions or
dispersion of DEHP in various vehicles may induce toxic re-
sponses when injected i.v. which may not occur when DEHP is
solubilized by having the ester migrate from PVC into blood.
Studies by Stern, et al. (1977) have indicated that the phar-
macokinetic 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 placed 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 volatility of most of the esters precludes them
from presenting an acute toxic response by inhalation. Gen-
erally, at least for the higher molecular weight phthalic
acid esters, only through heating will there be sufficient
vapor concentration to carry out an adequate inhalation
study.
Even though the phthalate esters have been in commercial
production for nearly 50 years, relatively few long-term tox-
icity studies appear in the literature. As vould be ex-
pected, subacute (or subchronic) studies are more plentiful
C-28 /
-------
but even these are few when one considers the large produc-
tion of these agents every year. Perhaps the meager toxico-
logical data can be attributed to the long use of these es-
ters 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 es-
ters can be seen in Table 7 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
two and eight percent in the diet produced only a minor
growth effect at the four and eight percent levels. At the
eight percent level, some indication of nephritic involvement
was detected. Dose levels less than eight percent showed no
such effect. A 90-day study in which the ester was applied
to the skin of rabbits led to an LD$Q 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 plas-
ticizer for cellulose 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 this ester
C-29
-------
TABLE 7
Calculated Allowable Daily Intake (ADI) for Various
Phthalate Esters (Krauskopf, 1973)
Maximum
Period No-Effect Level
Ester Species Days (mg/kg/day)
Di-2-ethylhexyl Rat 365 400
Rat 730 80
Dog 98 100
Rat 90 200
Dog 98 100
Rat 365 >60>200
Guinea pig 365 60
Dog 365 60
Dibutyl Rat 365 350-110
Rat 450 4.3
Diisonyl Rat 91 150
Dog 91 37
Heptyl nonyl Rat 90 60
Mouse 90 60
C-30
-------
without death occurring. Rabbits could be fed 3 ml/kg/day
without significant toxic response (Blickensdorfer and
Templeton, 1930). Diethyl phthalate does not act as a pri-
mary irritant when applied to the skin nor has it induced al-
lergic 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 respira-
tory tract.
Even though diethyl phthalate is not generally used as a
plasticizer in PVC tubings, Neergaard, et al. reported that
this ester was present in tubings used in hemodialysis equip-
ment and that the use of these tubings led to hepatitis in
several patients. When other tubings, presumably without di-
ethyl phthalate, were used the hepatitis did not occur. It
seems unlikely that the ester was responsible for the hepati-
tis 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 concentra-
tions of 0.01, 0.05, and 0.25 percent 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 ani-
mals grew normally as compared to the untreated controls.
Spasovski (1964) conducted a subacute inhalation study
lasting 93 days during which mice were exposed for six hours
a day to different concentrations of the ester. The concen-
trations ranged from 0.017 to 0.42 mg/1. Unfortunately, dur-
ing the study, the same animals received various exposure
C-31
-------
concentrations rather than specific concentrations for the
whole time period and thus interpretation of the results is
difficult even though Spasovski proposed a permissible stan-
dard 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 dose during the fourth and sixth weeks
of the experiments. The same group of animals also demon-
strated alterations in the phagocytic activity of neutrophils
after one month; these returned to normal. It is difficult
to conclude from this study the significance 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 behavioral changes vere
noted nor any weight loss discerned. The important finding
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/m3. Concen-
trations of 0.12 and 0.15 mg/m3 resulted in electrocortical
conditioned reflex in the three subjects in the study. When
the level was reduced to 0.093 mg/m3 no conditioned reflex
was noted. Men'shikova recommends a PSC value of 0.1 mg/m3.
C-32
7°
-------
Carter, et al. (1977) described a study on dibutyl
phthalate and the resultant testicular atrophy which oc-
curred. In the study, the ester was dissolved in corn oil
and administered orally (by intubation) for a period of time.
The dose administered was 2000 mg/kg while control animals
received corn oil in a volume of 5 ml/kg. The initial ef-
fect noted was a progressive reduction in weight of the
testes. In 14 days, the reduction amounted to 60 to 70 per-
cent 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 ap-
parently influenced zinc metabolism with an increase in the
excretion of zinc in urine. It was visualized that after
oral administration dibutyl phthalate is metabolized by non-
specific esterases in the gastrointestinal tract to the mono-
butyl 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 deficiency of zinc in testes tis-
sues is, according to the authors, the causative factor lead-
ing to the atrophized organ.
Milkov, et al. (1973) reported in 1969 that a group of
esters in an industrial environment produced various degrees
of toxic polyneuritis. These investigators studied 147
C-33
-------
persons (87 women and 60 men) the majority of whom were not
more than 40 years old. These industrial workers were ex-
posed primarily to dibutyl phthalate but other esters appar-
ently were also present but in much less concentrations.
These included dioctyl, diisooctyl and benzyl butyl phthal-
ates. Also, in some instances there were small amounts of
sebacates, adipates and tricresyl phosphate.
Until more occupational studies are performed, the re-
port by Milkov, et al. (1973) must be taken with some reser-
vation because of the presence of other chemical agents such
as tricresyl phosphate, an agent known for inducing poly-
neuritis.
Dibutyl (Diethylene Glycol Bisphthalate) (DDGB): Hall,
et al. (1966) studied the tcxicity of DDGB. They used a
commercial sample which also contained 15 percent dibutyl
phthalate and 5 percent (diethylene glycol) phthalate. The
oral LDjQ of this product in rats was found to be greater
than 11.2 g/kg and the i.o. 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 and 2.5 percent of the product, respec-
tively. Over the period of the study, there was a marked re-
duction 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
C-34
"V7
-------
percent level, oxaluria and hematuria were found in both
sexes, the oxaluria being assumed to be a direct consequence
of the in vivo liberation of diethylene glycol (a known pro-
ducer of oxalate stones in the bladder).
Butyl Benzyl Phthalate: Mallett and Von Hanun (1952) ad-
ministered 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 tox-
ic splenitis and degeneration of central nervous system tis-
sue with congestive encephalopathy. Further, myelin degener-
ation and glial proliferation were reported.
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
histopathological changes were not seen. The authors con-
cluded 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 toxicological data are available on it
than any of the other esters. It should be remembered that
DEHP is often used synonymously with the dioctyl phthalate
and, even though they are isomers, they have slightly dif-
ferent biological properties.
C-35
-------
The acute oral toxicity is very low, ranging from 14.2
to greater than 50 g/kg before lethality will occur in ro-
dents. Dermal absorption will occur but in rabbits approxi-
mately 25 ml/kg need to be applied to the skin to cause
death. Inhalation toxicity is also extremely low due to the
very low volatility of the ester.
In 1945, Shaffer, et al. (1945) reported a 90-day sub-
acute toxicity study in rats. Groups of animals were given
in feed 3.0, 1.5, 0.75 and 0.375 percent of the ester which
approximates daily intakes of 1.9, 0.9, 0.4 and 0.2 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, a slight decrease in growth was noted when compared
to the control animals. At the 3.0 and 1.5 percent doses,
tubular atrophy and degeneration in the testes were observed.
No deaths occurred in any of the treated animmals while blood
cell counts, hemoglobin concentrations and differential white
cell counts remained normal. The authors concluded that a no
adverse effect from oral administration would occur at ap-
proximately 0.2 g/kg/day or less while only a slight retarda-
tion 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 0.4, 0.13 and 0.04 percent of DEHP for
a maximum period of two years. In addition, a group of fil-
ial generation (F^) rats were given in feed 0.4 percent of
C-36
V'/
-------
DEHP for one year. Control groups of rats were maintained on
the same basic diet without the ester. The investigators ex-
amined the following signs and symptoms of toxicity: mortal-
ity, life expectancy, body weight, food consumption, liver
and kidney weights, micropathological changes, neoplasm, he-
matology and fertility.
Over the two-year period for the PJ group and over a
one-year period for the F^ group, a number of deaths oc-
curred ,. However, these deaths were not attributed to the es-
ter since they were also noted in the control animals.
The mean liver and kidney weights, as percentage of body
weights, were found to be increased over those of the con-
trols in both the initial group (PI) and their offspring
(FI> which had received the diet containing 0.4 percent
DEHP. The results were statistically significant. Histo-
pathological examination of the liver and kidney tissues of
treated animals did not reveal statistically significant dif-
ferences from organs of control animals. The authors did
suggest that even though pathological changes in the two or-
gans of treated groups were not different from control ani-
mals, the increase in size of the organs may indicate a toxic
response. Results from 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
0.13 and 0.04 percent DEHP for one year. Similar criteria,
C-37
'H
f \
-------
with the exception of hematology and fertility, as used in
the rat study were employed. Liver weights, as percentage 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 concentra-
tions 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 admin-
istered. 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 ef-
fect, confirmed the results of Carpenter, et al. (1953). A
chronic oral toxicity study in male and female rats was con-
ducted in which groups of animals received in their feed 0,
0.1 and 0.5 percent DEHP. At various time periods, rats were
sacrificed and food consumption, body weight, and liver,
testes, kidneys, lungs, brain, stomach, heart and spleen
weights recorded. Histopathological studies were also con-
ducted on selected tissues and organs. The study was termi-
nated after 24 months. Significant increases in liver and
kidney weights were noted at the 0.5 percent dose level for
C-38
-------
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 pa-
thology 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 within three months when a dog was administered
5 g/kg/day of DEHP but not with 0.1 g/kg/day. The small num-
ber 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 associated 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 LD50
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. These
included: (1) mice injected for at least ten weeks, and (2)
the apparent L?$Q remained constant for three consecutive
C-39
-------
weeks. DEHP and DOP were included in the list of esters
studied. The first week, the LD5Q for DEHP was 38.35 ml/kg
and 67.18 ml/kg for DOP. The second week, the LD5Q was re-
duced to 6.40 ml/kg for DEHP and 25.51 ml/kg for DOP. By the
end of the 12th week, the LD5Q was reduced to 3.09 ml/kg for
DEHP and to 1.37 ml/kg for DOP. A cumulative toxicity factor
was calculated for each of the esters (acute LD5Q/chronic
LDso) and for DEHP this value was 27.99 (indicating that the
toxicity had increased by this factor). A similar calcula-
tion for DOP 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 factor for both DEHP and DOP is not clear and the
reasons for these results, when compared to the other esters,
are presently not explainable. It is possible to speculate
that very high exposure doses prevent the body from eliminat-
ing 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 re-
sults, since this type of administration 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.
C-40
-------
These investigators, however, could not find light micro-
scopic evidence of injury to these organs using histopatho-
logical 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 2000
mg/kg/day for periods of 4, 7, 14, and 21 days. Control ani-
mals received 0.5 ml/100 g body weight of the vehicle. The
investigators noted relative liver weight increased progres-
sively 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 activities and/or
levels determined (for each of the time periods): succinate
dehydrogenase, aniline 4-hydroxylase, biphenyl 4-hydroxylase,
glycose-6-phosphatase, cytochrome P-450? protein contents,
and alcohol dehydrogenase. Alcohol dehydrogenase activity
and microsomal protein and cytochrome P-450 contents in- .
creased markedly initially but then decreased during the time
of treatment. On the other hand, microsomal glucose-6-phos-
phatase, 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
dilation of the smooth endoplasmic reticulum and swelling of
the mitochondria.
041
-------
Lake, et alo (1975) studied the monoester and found that
liver changes in treated rats closely resembled those pro-
duced by DEHP0 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
1000 and 1500 ppm of i4C»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. Re-
maining animals were returned to a normal diet and sacrificed
at intervals during the subsequent two to three weeks and
tissues prepared for analysis,, At the 5000 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. Electron microscopy of liver tissue revealed only a
slight increase in the amount of smooth endoplasmic reticu-
lum. Returning animals to a normal diet resulted in liver
weight returning to normal „ There was no apparent change in
liver weight in those animals receiving the 1000 ppm DEHP.
Additional studies by these authors did not reveal the ac-
cumulation of DEHP j.n body organ tissues.
Nikonorow,, et alo (1973) reported that a daily dose
level of 0.35 percent (in feed) of DEHP 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.
042
-------
Kevy, et al. (1978) studied the toxic effects of DEHP
solubilized in monkey blood or blood products by storing the
animal blood (or blood product) in PVC blood bags. These
products were then transfused into the animals for time
periods ranging from six months to one year. This dosing
program attempted to mimic actual transfusion levels expected
in selected patients requiring large-volume blood or blood
products. The total concentration 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 technique 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 per-
sisted. DEHP was also found in liver tissue in treated ani-
mals many months after the last transfusion. The work of
Kevy and 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 fre-
quency of exposure, and the health status of the patient.
Biochemical studies on rat blood and liver at 21 days
after injection of 5 ml/kg DEHP i.p. on days one, five and
ten produced the following results: a decrease in the activ-
ity of succinic dehydrogenase and an increase in alkaline
phosphatase activity in the liver; serum enzyme values were
not 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.
043
-------
Though it is recognized that different routes and dosage
forms will alter the pharmacokinetic disposition of com-
pounds, DEHP from several different routes (oral, i.p., i.v.)
can produce hepatotoxic responses depending upon the specific
dose and the frequency of exposure.
Seth, et al. administered i.p. 5 ml/kg of DEHP (undi-
luted) to 10 male and 20 female rats on days one, five and
ten. On the 22nd day of the study, all animals were sacri-
ficed and one test is or ovary was removed and retained for
enzymatic studies. A control group of rats received an equal
volume of saline. Results of the study demonstrated that the
scrotums in all animals were enlarged but no gross abnor-
mality was discerned. Succinic dehydrogenase (SDH) and ade-
nosine triphosphatase (ATPase) activities were significantly
reduced while that of ^-glucuronidase was increased in both
organs of the test animals. Histopathologic examination of
the testes of the animals revealed degenerated tubules show-
ing marked vacuolization of the cytoplasma of spermatogonial
cells and eccentric nuclei. No apparent alterations (histo-
pathologic) were noted in the ovaries of the DEHP 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,, Whan 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.
C-44
-------
Even though mention was made that other esters of phthalic
acid were studied no data were presented. Thus, the reader
may assume that these other esters did not have the same
\
toxic properties to testes as either DEHP or the dibutyl
ester. It seems possible that DEHPf like dibutyl 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 inhibition of hepatic sterolo-
genesis, inhibition of fatty acid oxidation by heart mito-
chondria, stimulation of fatty acid oxidation by hepatic
mitochondria, and an ability to modify the pattern of circu-
lating plasma lipoproteins. In several of the studies, rab-
bits and pigs were also used and led to the conclusion that
the response of mammalian tissues to phthalate esters is var
iable depending upon the species. The toxic implications of
alteration in lipid metabolism to man is 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, suggest-
ing that the metabolites must play the major factor in pro-
ducing a toxic response. It also appears that man, rat,
baboon and ferret may handle DEHP as well as other esters in
a similar manner (Lake, et al. 1977).
C-45
-------
Synergism and/or Antagonism
There are no data available on the synergism or
antagonism of phthalate esters.
Teratogenicity
Singh, et al. (1975) included eight phthalic acid esters
in a rat teratogenic study. The esters included the follow-
ing: dimethyl, dimethoxyethyl, diethyl, dibutyl, diisobutyl,
buty carbobutoxymethyl, dioctyl and di-2-ethylhexyl phthal-
ates. For all the esters, except two, the dose administered
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 in-
cluded: untreated rats, rats treated with 10 mg/kg of dis-
tilled water, rats treated with 10 ml/kg of normal saline and
rats treated with 10 ml/leg 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 numbeir of corpora lutea, resorption
sites, and viable and dead fetuses. Additionally, both vi-
able and non-viable fetuses were excised, weighed, and ex-
amined for gross malformation. From 1/3 to 1/2 of the
fetuses, using those which showed no gross malformation when
possible, were prepared as transparent specimens to permit
visualization of skeletal deformities.
C-46
-------
All of the esters produced gross* or skeletal abnormal-
ities which were dose related. The most common gross abnor-
malities in the treated animals were absence of tail ano-
phthalmia, twisted hands and legs, and hematomas. Skeletal
abnormalities included elongated and fused ribs (bilateral
and unilateral), absence of tail bones, abnormal or incom-
plete skull bones, and incomplete or missing leg bones. Dead
fetuses were found in the groups treated with dimethyl, di-
methoxyethyl and diisobutyl phthalates. The most embryotoxic
agent in the series was dime thoxyethyl phthalate. Each of
the esters also reduced the weight of the fetuses when com-
pared to the controls. 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 terato-
genic effects if the compounds had been administered orally
or by other routes.
In another study by Peters and Cook (1973), pregnant
rats were administered 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 DEHP such as excessive bleeding, incomplete
expulsion of fetuses and maternal deaths. Teratogenic
studies on dibutyl and dimethyl phthalates were also con-
ducted by these authors, but the adverse effects were less
C-47
-------
than those observed for the DEHP-treated rats. It was inter-
esting 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 in-
jected i.p. with labeled di-2-ethylhexyl phthalate and di-
ethyl phthalate. The results demonstrated that these phthal-
ates could pass through the placental barrier suggesting 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, com-
mercial phthalate esters in chick embryos. They found that
dibutyoxyethyl phthalate, di-2-methoxyethyl phthalate and
octyl isodecyl phthalate produced damage to the central ner-
®
vous system of the developing chick embryo when compared to
control embryos receiving an oil and to an untreated group.
In a study reported by Nikonorow, et £l. (1973), preg-
nant rats were administered orally 0.34 and 1.70 g/kg/day of
DEHP during the gestation period. Another series of rats re-
ceived 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 statis-
tically significant reduction in fetus weight at both dose
levels for DSHP but only at the higher dose level for the
dibutyl phthelateo The number of resorptions were noted for
DEHP at bcth dose levels but only at the higher dose level
for dibutyl phthalate. No detectable differences were
C-48
-------
observed in the number of sternum ossification foci, develop-
ment 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 hu-
mans on a daily basis is extremely small as compared to the
doses used in the previous studies, it seems remote that
teratogenic effects would be produced in humans. Further
studies in which the esters are administered orally to preg-
nant females should, however, be carried out to verify this
assumption.
Mutagenicity
Studies of the effect of phthalic acid esters on genetic
changes in animals are not adequate 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 dimethoxy-
ethyl phthalate (DMEP) in a study on the mutagenic and anti-
fertility effects in mice. The experiment followed the gen-
eral procedure used in conducting the dominanat 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 mlAg), and 2/3 (25.6 ml/kg) of
the LD5Q. A similar dose pattern was used for the DMEP or
1/3 (1.19 ml/kg), 1/2 (1.78 mlAg) and 2/3 (2.38 ml/kg) of
the LD5Q.
C-49
-------
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 pregnan-
cies occurred. Fewer effects were noted at the lower dose
levels. DEHP appeared to have a more persistent 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-ethyl-
hexyl and butyl benzyl phthalate as well as phthalate acid.
Positive responses were found for the dimethyl and diethyl
phthalates. The remaining compounds were found to be non-
mutagenic under the test conditions.
Studies by Turner, et al. (1974), showed the DEHP did
not produce genetic damage in lymphocytes but did inhibit
mitosis and growth.
It is clear that more studies on the mutagenic effects
must be conducted before a definite conclusion can be made
concerning the risk of a population exposed to the phthalate
C-50
-------
esters. The antifertility effect appears to be much stronger
and the question which still needs to be answered is what ef-
fects would lower doses have upon males repeatedly exposed to
these esters. Epidemiological evidence on this subject is
lacking, and thus human risks cannot accurately be por-
trayed.
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 suggest-
ing but not proving that these compounds may have a cancer
liability. A long history of use of both of these compounds,
however, has not implicated these as even weak carcinogenic
agents. It would appear, however, that consideration should
be given to cancer studies of these two esters in animal
models to ensure that a potential cancer threat does not
exist.
Other Biological Effects
Cellular Toxicity: In recent years, a number of in
vitro tests have become useful in assessing the toxicity of
chemicals. Even though the results may net-always be extra-
polated to animals or humans, the proper in vitro system can
generate very useful data which can assist in determining the
toxic consequences of a chemical. Tissue and organ culture
methods are now widely used toxicity testing methods.
C-51
-------
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 liquids, 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 liquids, 35 mg
of the ester were placed on a paper disk which was previously
placed on the agar. After 24 hours of incubation, the cells
were examined for cytotoxicity. Table 8 includes the results
of the screening tests. In the same table are the results
from a mouse t.oxicity 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 molecu-
lar 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 con-
centration of 0.18 mM, which is equivalent 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 DEHP concentration was reduced to 0.10 mM which is com-
parable to the concentration found in whole blood stored at
4°C for 14 days.
C-52
/•
r
-------
TABLE 8
Results of the Toxicity Evaluation of Phthalate Esters
on the Mammalian Cell Cultures and Mice
Phthalates
Chick
Embryo
Cells
L-Cells
Mice
CH-
n-C3H7
n-C4H9
iso—C4
n-C5Hu
Cyclo-CsHg
n-C6H13
n-C7H15
Cyclo-C
n-C8H17
n-C10H2i
n-c12H25
From: Nematollahi, et al. 1977.
Note: In tissue culture test: + indicates cytotoxic; - in-
dicates noncytotoxic; +_ indicates questionable results
In mouse test: + indicates 2 or 3 deaths; - indicates
no deaths; _+ indicates only one death.
C-53
-------
In another tissue culture study, Jones, et al. (1975)
reported the 1050 (concentration required to inhibit cell
growth by 50 percent) on a number of phthalic acid esters.
The IDso values are shown in Table 9. As will be noted from
the table, IDso for DEHP came to 70 uM. In comparing this
IDSO 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), indicated the actual solubility in the
medium. The 70 uM solubility concentration would be approxi-
mately 0.05 mM which is in line with the Jacobson value con-
sidering that slightly different techniques were employed.
The most cytotoxic ester in the series was butyl glycolyl
butyl phthalate.
The IDso dose for a group of phthalate esters has been
reported for mouse fibroblasts in cell culture (Autian,
1973). These values are included in Table 1C. It is inter-
esting to note that the most cytotoxic agent in the series
was DEHP, an agent having a very low order of acute toxicity
in animals and man. As can be seen from the table, the tox-
icity of these compounds in general increased as the molec-
ular weight increased.
A report by Dillingham and Autian (1973), indicates that
dimethyoxyethyl phthalate is much more toxic to mouse fibro-
blast cells undergoing significant rates of cell division
than nonreplicating cells. This observation suggests that
any tissue which undergoes periodic increases in protein
C-54 j
-------
TABLE 9
1050 Values for a Series of Phthalate Esters
Using WI-38 Cells
Agent
(Phthalate)
Di-n-butyl
Di-iso-butyl
Dime thoxy ethyl
Butyl glycol butyl
Di-n-octyl
Di-2-ethylhexyl
Molecular
Weight
278
278
282
336
391
391
ID50
(UM)
135
85
3500
12
170
70
Solubility
(mol/liter)
0.008
Very Low
0.040
Very Low
Very Low
Very Low
Taken in part from Jones, et al. 1975.
TABLE 10
1050 of a Group of Phthalic Acid Esters in Tissue Culture
(Mouse Fibroblasts)
Ester
Dimethyl
Diethyl
Dibutyl
Dime thoxy ethyl
Di-2-ethylhexyl
Molecular
Weight
194
222
278
282
390
Water Sol.
(mole/1)
0.0263
0.0048
0.008
0.0400
0.0004
ID
I°50
0.007
0.003
0.0001
0.0084
0.00005
Taken in part from Autian, 1973,
C-55
-------
turnover related to changes in cell division rate and meta-
bolic activity (protein synthesis) may increase the suscepti-
bility of these cells to the toxic effects of phthalic es-
ters. Thus, it is possible that the teratogenic and embryo-
toxic 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 meta-
bolic activity in contrast to somatic cells which have a much
lower rate of cell division and metabolism of the somatic
tissue.
Kasuya cultured cerebella from newborn rats and tested
three phthalate esters (dimethyl, diethyl and dibutyl phthal-
ates). Various concentrations of each of the esters were
dissolved in calf serum and then added to the cells. The
overall toxicity to the cells was in the following order:
DBP>DEP>DMP. As will be noted, the toxicity of the three
esters increased with molecular weight similar to cell cul-
ture results reported by Dillingham and Autian.
At a concentration of 4 vg/ml in tissue culture media,
DEHP produced complete cessation of beating chick embryo
heart cells maintained in tissue 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
i
been concern that DEHP, wheA extracted from medical devices
such as blood bags and tubipgs, might have a deleterious
056
-------
effect upon blood components and also lead to the syndrome
referred to as "shocked lungs." DEHPf solubilized with a
surfactant and injected i.v. in rats, produced lung involve-
ment 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 "non-toxic" effect
while DEHP solubilized with a surfactant produced a toxic
effect.
Rubin (1975) reported that DEHP, solubilized with a sur-
factant and injected 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 experi-
ment (Rubin, 1976). Control rats, treated in a similar man-
ner but not receiving any DEHP, did not demonstrate the toxic
lungs.
Herman, et al. (1977) conducted studies in which rats
were administered blood or blood components, previously in
contact with PVC strips, to detect the effect DEHP (extracted
from the plastic) would have on lung tissue. ACD-preserved
C-57
-------
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 DEHP. After' storage for two
weeks, 0.5 ml of blood were administered 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
CPD-preserved rat or human blood after storage in glass alone
or in glass containing PVC strips and/or PVC enriched with
DEHP. Concentration cf 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 DEHP.
Evans Blue was used as an indicator to detect the per-
meability of excised lung tissue. Animals given ACD-pre-
served blood which had contact with PVC demonstrated an in-
creased permeability when compared to control animals.
Administration of platelet-rich and platelet-poor plasma
showed no significant increase in lung permeability. CPD-
preserved blood in contact with the plastic strips showed an
increased permeability which was greater than the CPD 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 en-
riched with DEHP showed variable degrees of septal thicken-
ing, perivascular edema and perivascular accumulation of
mononuclear cells when compared to lungs of control rats.
The authors suggest that blood-plastic contact during storage
c-sa
-------
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 liquid particles 'of DEHP which, in
turn, can be administered to humans (Needham and Luzzi,
1973). Depending upon the size-frequency of 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 ug/ml. Eight of the plasma
samples 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 ug/nil.
Fractionated proteins 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 taking place in the product. They indi-
cate that when measuring DEHP content of blood and blood pro-
ducts 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
C-59 .> /
-------
butyl glycolyl butyl phthalate. These esters were also emul-
sified with acacia and injected at 250 mg/kg and 500 mg/kg
dose levels. After 30 minutes, hexobarbital solution was ad-
ministered i.p. A significaant 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 em-
ployed 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 sensitivity
to hexobarbital nor an alteration in rate of hexobarbital
metabolism by the liver, but to the effect of DEHP in the
distribution of hexobarbital into various organs.
Swinyard, et al. (1976) also found an increase in hexo-
barbital sleeping time from DEHP. It was interesting to note
that olive oil also produced an increased sleeping time simi-
lar to DEHP. These authors concluded that the effect of DEHP
was nonspecific due to the physical characteristic of the es-
ter 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 DEHP 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.
C-60
-------
From the information available, it is clear that DEHP
prolongs the sleeping time of short-acting barbiturates. In
the instance of acute studies, the cause of the prolongation
of sleeping time may, in fact, be due to nonspecific factors,
probably to the lipophilic reservoir mechanism advocated by
Swinyard, et al. (1976). On the other hand, repeated pre-
treatments with DEHP may have an effect upon the liver and
enzyme systems. Since liver involvement has been noted by
several investigators ir. subacute toxicity studies in rats
and monkeys, the DEHP may, in these cases, be producing a
specific toxicological effect.
C-61
-------
CRITERION FORMULATION
Existing Guidelines and Standards
The Threshold Limit Value for dimethyl, dibutyl and di-
2-ethylhexyl phthalate esters established by the American
Conference of Governmental and Industrial Hygienists is 5
mg/m^.
The Food and Drug Administration has approved the use of
a number of phthalate esters in food packaging materials.
Prior to 1959 (before enactment of the food additive amend-
ment), FDA approved five esters. These are: diethyl phthal-
ate, diisobutyl phthalate, ethyl phthalyl ethyl glycolate,
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 throuoh a number of routes such as industrial sites in
which the esters are manufactured or used. Esters may also
reach man through indirect means such as inhalation of the
esters inside vehicles containing PVC products firom foods and
from water. Direct injection i.v. of specific phthalate es-
ters can also occur when PVC blood bags and tubings are used
to transfuse blood and blood products to man. The ubiquitous
C-62
-------
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 ex-
posed to these esters.
Even though it is well established that workers in occu-
pations in which phthalate esters are used are exposed to
various levels of phthalate esters and thus can absorb these
esters through inhalation or through dermal absorption, 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 (mosquito 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 tha Bureau of Foods (FDA) in
1974 to determine if phthalate esters were entering the food
supply through the processing, packaging, handling and trans-
portation chain. In the study, ten basic and stable food
products were analyzed for the presence of these esters.
Conclusions reached in the report are presented below:
1. The frequency and levels of phthalate esters re-
ported as well as the possible cumulative intake
of phthalates in baked beans in cans or jars, can-
ned 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 commodities. Dibutyl phthalate, dicyclohexyl
phthalate and butylphthalylbutyl glycolate were
found in comparatively few samples. Diisoctyl and
diisodecyl phthalates, although looked for, were
not detected.
C-63
-------
3. Phthalate ester contamination was found in a higher
proportion of milk and cheese samples than in
other foods. (However, the findings are uncer-
tain. )
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 levels of esters were 22.8
and 24.9 ppm 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), informa-
tion is presented dealing with phthalate (DEHP and DNBP)
residues in various commercial foodstuffs in Japan. They
concluded that foods packaged in plastic films with printing
are a greater source of contamination to the product with the
esters than if the foods were in plastic bottles. They also
noted that persons had significantly higher levels of the
esters after meals from foods packaged in the film. Ex-
tremely 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 con-
clusion 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."
C-64
-------
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 quan-
tity 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 es-
ters. These are workers in the industrial environment in
which the phthalates are manufactured or used and patients
receiving chronic transfusion of blood and blood products
stored in PVC blood bags<=
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 i0p=y the esters can act as terato-
genic agents and possibly as mutagenic agents in rats. These
esters also have an effect upon gonads in rats. Evidence is
also at hand to show that the esters may bring about biochem-
ical and pathological changes in the liver of rats when .re-
peatedly administered orally or by i.p. When solubilized in
blood components, DEHP has demonstrated liver involvement
when these products have been repeatedly administered i.v. to
monkeys. Inhalation studies in rats and man suggest that
C-65
-------
certain phthalates may be responsible for neurological dis-
orders, but these results need further verification since
other non-phthalate esters may also have been present leading
to the problems.
Since a number of phthalate esters are in the environ-
ment 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 reported to
establish an allowable daily intake (ADI). In calculating
the API, an uncertainty factor of 100 was used based upon a
70 kg person. Table 11 taken from Shibko (1974), lists eight
esters in which the "no effect" dose was established from
chronic toxicity studies in rats or dogs. The table also
includes the number of days the animals were fed the specific
phthalate esters and the calculated ADI. It will be noted
that the ADI ranged from a low of 9.8 mg/day for dicyclohexyl
phthalate to a high of 700 mg/day for dimethyl phthalate.
For the sake of establishing water quality criteria, it
is assumed that on the average a person ingests 2 liters of
water and 18.7 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. Biomagnification factors for dimethyl, diethyl,
dibutyl and di-2-ethylhexyl esters were derived by the U.S.
E?A ecological laboratories, Duluth (see Inge$tion from
Foods).
C-66
-------
TABLE 11
Calculated Allowable Daily Intake in Water and Fish
for Various Phthalate Esters
1.
2.
3.
4.
n
a\
~~> 5.
6.
7.
8.
**
***
Ester
Dimethyl
Diethyl
Dibutyl
Dicyclohexyl
Methyl phthalyl
ethyl glycolate
Ethyl phthalyl .
ethyl glycolate
Butyl phthalyl
ethyl glycolate
Di-2-ethyhexyl
No Effect Dose*
(mg/kg/day)
1000
625
18
14
750
250
140
60
From: Shibko, 1974.
Allowable Daily Intake for 70 kg
F = Biomaanif ication factor.
Species
Rat
Dog
Dog
Dog
Rat
Rat
Dog
Dog
person (100 s
Days
104
52
52
52
104
104
104
52
safety
ADI**
(mg/day)
700
438
12.6
9.8
525
175
98
42
factor) .
F*** Recommended
Criteria
mg/1
130 160
270 60
26 5
Not
Established
Not
Established
Not
Established
Not
Established
95 10
-------
Due to lack of data, bioconcentration factors could not be
derived for dicyclohexyl, methyl phthalyl ethyl glycolate,
ethyl phthalyl ethyl glycolate and butyl phthalyl ethyl
glycolate.
The equation for calculating an acceptable amount of
ester in water based on ingestion of 2 liters of water and
18.7 g fish is:
(2/1) X + (0.0187 x F) X = ADI
where 2/1 = 2 liters of drinking water consumed
0.0187 kg = amount of fish consumed daily
F = biomagnification factor
ADI = Allowable Daily Intake (mg/day for 70 kg person)
For example, consider that the ADI for dimethyl phthalate is
700 mg/day and the biomagnification factor is 130, the above
equation can be solved as follows:
2X + (0.0187 x 130)X = 700
2X + (2.43)X = 700
4.43X - 700
X = 158 (or~160 mg/1)
Thus, the recommended water quality criterion is 160
mg/1.
Similar calculations were made for each of the esters
and are presented below:
Diethyl
2/1 X + (.0187 x 270)X = 438
2X + 5.05X = 438
7.05X = 438
X = 62 mg/1 (cr*-»60 mg/1)
C-68
Iti-
-------
Dibutyl
2/1 X + (.0187 x 26) = 12.6
2X + .468X = 12.6
2.468X = 12.6
X = 5.10 mg/1 (or— 5 mg/1)
Di-2-ethyhexyl
2/1 X + (.0187 x 95) = 42
2/1 + 1.7765 = 42
3.7765X » 42
X = 11.12 mg/1 (or ^ 10 mg/1)
Thus, the recommended water quality criteria for four
phthalate esters are:
dimethyl 160 mg/1
diethyl 60 mg/1
dibutyl 5 mg/1
di-2-ethylhexyl 10 mg/1
(see Table 11).
It seems clear that exposure from the water route pre-
sents no real risk to the population in regard to the phthal-
ate esters. Reported levels of phthalate esters in U.S.
surface waters have only been in the ppb range, at approxi-
mately 1 to 2 ug/1 (see Ingestion from Water section).
Other routes of exposure such as inhalation (industrial
sites manufacturing 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
C-69 {
\
;) .'c'7
-------
and milk may, on some occasions, reach 50 ppm. Also a spe-
cial group at risk will be patients to whom chronic trans-
fusions of blood and blood products are administered.
Although it is recognized that routes of exposure other
than water contribute 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 con-
tent 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 established. The percent contri-
bution of drinking water and of ingesting contaminated fish
is given in the following table. Also given are the criteria
levels recommended if exposure is assumed to be from fish and
shellfish products alone.
C-70
-------
Esters
Criteria level
mq/1
% Contribution
of drinking water
% Contribution
of Fish Products
Criteria if Exposure
is from Fish Alone
mq/1
Dimethyl
Diethyl
Dibutyl
D-2-ethylhexyl
160
60
5
10
45
29
81
53
55
71
19
47
288
87
26
24
o
-J
-------
REFERENCES
Albro, P.W.f et al. 1973. Metabolism of diethhexyll phthal-
ate by rats. Isolation and characterization of the urinary
metabolies. Jour. Chromatogr. 76: 321.
Autian, J. 1973. Toxicity and health threats of phthalate
esters: Review of the literature. Environ. Health Perspect.
June 3.
Bell, P.P. 1976. Inhibition of hepatic sterol and squalene
biosynthesis in rats fed di-2-ethylhexyl phthalate. Lipids.
11: 769.
Bell, P.P., and D. Nazir. 1976. Effect of dietary di-2-
ethylhexyl phthalate on lipid biosynthesis in selected tis-
sues from the rat, in vitro. Lipids. 11: 216.
Bell, P.P., and P.J. Gillies. 1977. Effect of dietary di-2-
ethylhexyl phthalate on oxidation of l4C-palmitoyl CoA by
mitochondria from mammalian heart and liver. Lipids. 12:
581.
Bell, P.P., et al. 1976. Studies on lipid biosynthesis and
cholesterol content of liver and serum lipoproteins in rats
fed various phthalate esters. Lipids. 13: 66.
C-72 •"
\
-------
Bell, P.P., et al. 1978. Effect of phthalate esters on
serum cholestrol and lipid biosynthesis in liver, testes and
epididymal fat in the rat and rabbit. Lipids 13: 673.
Berman, I.R., et al. 1977. Pulmonary effects of blood con-
tainer materials. Surgical Forum. 28: 182.
Blickensdorfer, P., and L. Templeton. 1930. A study of the
toxic properties of diethyphthalate. Jour. Am. Pharm. Assoc,
19: 1170.
Bower, R.K., et al. 1970. Teratogenic effects in the chick
embryo caused by esters of phthalic acid. Jour. Pharmacol.
Exp. Therap. 171: 314.
Braun, B., and H.J. Kummell. 1963. The use of plastic con-
tainers for storing blood and transfusion solutions. Dtsch.
Apotheker-Zig. 103: 467.
Carpenter, C.P., et al. 1953. Chronic oral toxicity of
di(2-ethylhexyl) phthalate for rats, guinea pigs and dogs.
AMA Arch. Ind. Health 8: 219.
Carter, B.R., et al. 1977. Studies on dibutyl phthalate-
induced testicular atrophy in the rat: Effect on zinc
metabolism. Toxicol. Appl. Pharmacol. 41: 609.
C-73
-------
Corcoran, E.F. 1973. Gas-chromatographic detection of
phthalic acid esters. Environ. Health Perspect. Jan. 13.
Cordle, F., et al. 1978. Human exposure to polychlorinated
biphenyls and polybrominated biphenyls. Environ. Health
Perspect. 24: 157.
Corley, J.H., et al. 1977. Effect of various factors on the
amount of plasticizer in intravenous solutions packaged in
flexible bags. Am. Jour. Hosp. Pharm. 34: 259.
Daniel, J.W., and H. Bratt. 1974. The absorption, metabolism
and tissue distribution of di(2-ethylhexyl) phthalate in
rats. Toxicology 2: 51.
Darby, T.D., and R.K. Ausman. 1974. Particulte matter in
polyvinyl chloride intravenous bags. New England Jour. Med.
290: 579.
Dillingham, E.G., and J. Autian. 1973. Teratogenicity, muta-
genicity and cellular toxicity of phthalate esters. Environ.
Health Perspect. Jan. 81.
Draize, J.H., et al. 1948. Toxicological investigations of
compounds proposed for use as insect repellents. Jour.
Pharmacol. Exp. Ther. 93: 26.
C-74 f
-------
Dvoskin, I.A.G., et al. 1961. Hygienic assessment of certain
polymers (provinols). (Translated title). Mosk. Nauchn.
Inst. Gigieny. No. 9: 105.
Gaunt, I.F., et al. 1968. Acute (rat and mouse) and short-
term (rat) toxicity studies on dialkyl 79 phthalate. Food
Cosmet. Toxicol. 6: 609.
Golberg, L. 1966. Liver enlargement produced by drugs: Its
significance. Proc. Eur. Soc. Study Drug Tox. 7: 171.
Guess, W.L., et al. 1967. A study of polyvinyl chloride
blood bag assemblies. I. Alteration or contamination of ACD
solutions. Drug Intelligence. 1: 120.
Hall, D.E., et al. 1966. Acute (mouse and rat) and short-
term (rat) toxicity studies on dibutyl (diethylene glycol
bisphthalate). Food Cosmet. Toxicol. 4: 383.
Harris, R.S., et al. 1956. Chronic oral toxicity of 2-ethyl-
hexyl phthalate in rats and dogs. AMA. Arch. Ind. Health
13: 259.
Hillman, L.S., et al. 1975. Identification and measurement
of plasticizer in neonatal tissues after umbilical catheters
and blood products. New England Jour. Med. 292: 381.
C-75
-------
Bites, R.A. 1973. Phthalates in the Charles and Merrimack
Rivers. Environ. Health Perspect. Jan. 17.
Jacobson, M.S., et al. 1974. The toxicity of human serum
stored in flexible polyvinyl chloride containers on human
fibroblast cell cultures: An effect of di-2-ethylhexyl
phthalate. Res. Commun. Chem. Pathol. Pharmacol. 9: 315.
Jacobson, M.S., et al. 1977. Effects of a plasticizer
leached from polyvinyl chloride on the subhuman primate: a
consequence of chronic transfusion therapy. Jour. Lab. Clin.
Med. 89: 1066.
Jaeger, R.J., and R.J. Rubin. 1970. Plasticizers from plas-
tic devices: Extraction, metabolism, and accumulation by
biological systems. Science 170: 460.
Jaeger, R.J., and R.J. Rubin. 1972. Migration of a phthalate
ester plasticizer from polyvinyl chloride blood bags into
stored human blood and its localization in human tissues.
New England Jour. Med. 287: 1114.
Jones, A.E., et al. 1975. Phthalate ester toxicity in human
cell cultures. Toxicol. Appl. Pharmacol. 31: 283.
Kasuya, M. Toxicity of phthalate esters to nervous tissues in
cultures. Report from Dep. Pub. Health, Sapporo Medical
College, Sapporo, Japan (in English).
C-76
'I
-------
Revy? SoVoj, et al» 1978= Toxicology of plastic devices hav-
ing contact with blood „ Rep0 NOIL HB 5-2906., Natl. Heart,
Lung and Blood J.nsto Bathesdaj, Md0
Krauskopf, L0G0 IS7Iic Studies on the toxicity of phthalates
via ingestiono Envircn0 Health Perspectc Jan. 61.
Lake,, BoGo, et alo 19750 Stuidies on the hepatic effects of
orally administered dx-(?,-ethylhexyl) phthalate in the rat.
Toxicolo Applo Pharmaco.lo 32s 355 0
Lake? BoGOI7 at alo 1S770 xae in vitro hydrolysis of some
phthalate diastars by hepatic and intestinal preparations
from various species „ Tojs.icc.lo Appl0 Pharmacol. 39: 239.
Lawrence^, W,Hop et alo r.9730 A lexicological investigation
of some acute y short-tsr.'m and chronic effects of administer-
ing di-2-ethylhexyl phthalafc© JDHHP) and other phthalate
esters o Environ „ Res0 Ss ":. 0
Mallette^, FoS.,, and Eo Von Eeastio 13520 The toxicity and skin
effects of compounds used in the rubber and plastics indus-
tries,, II o Plasticizsz'So &rch° Indo Kyg» Occup. Med. 6: 231.
Mayer „ P0Lo 197S, Residua dynamics of di-2-ethylhexyl
phthalate in fathead minnows (Pimephal.£s promelas) . Jour.
Board Can0 33s ?,S100
C-77
-------
fien'shikova, T.A. 1971. Hygienic evaluation of dibutyl
phthalate in relation to the use of polymeric materials for
finishing living quarters on ships. (Translated title).
Gig. Sanit. 36: 23.
\
Meyler, F.L., et al. 1960. The influence of polyvinyl
chloride tubing on the isolated perfused rat's heart. Circ.
Res. 8: 44.
Milkov, L.E., et al. 1973. Health status of workers exposed
to phthalate plasticizers in the manufacture of artificial
leather and films based on PVC resins. Environ. Health
Perspect. Jan. 175.
Needham, T.E. Jr., and L.A. Luzzi. 1973. Particulate matter
in polyvinyl chloride intravenous bags. New England Jour.
Med. 289: 1256.
Needham, T.E. Jr., and R.D. Jones. 1978. Delivery of
plasticizer from standard intravenous-administration sets.
New England Jour. Med. 299: 1472.
Nematollahi, J., et al. 1977. Plasticizers in medical
application. I, Analysis and toxicity evaluation of dialkyl
benzene-dicarboxylates. Jour. Pharm. Sci. 56: 1446.
C-78
-------
Mkonorow, M, , et al. 1973. Effect of orally administered
plasticizers and polyvinyl chloride stabilizers in the rat.
Toxicol. Appl. Pharmacol. 26s 253.
Peakall, D.B, 1975= Phthalate esters; Occurrence and bio-
logical effects. Residue Rev» 54: 1.
Peters, J.W., and R.M. Cook. 1973= Effects of phthalate
esters on reproduction of rats. Environ. Health Perspect.
Jan. 91.
Pfab, W. 1967= Migration of phthalate plasticizers from
lacquered aluminum foils on fatty foods*, (Translated title),
Deut. Lebensm.-Rundsch. 63s 72.
Rubin R.J« 1975o Metabolism and acute lung toxicity of solu-
bilized di(2-ethyhexyl) phthalate (DEHP) in rats. Page 205
in Proc. Sixth Int. Congr. Pharmacol. Helsinki, Finland.
Vol. 6. Mechanism of toxicity and metabolism.
Rubin, R.J. 1976o Transcript of proceedings. Workshop on
adenine and red cell preservation. Food Drug Admin. Bur.
Biol. Dep. Health Edu. Welfare.
Rubin, R.J., and R.J. Jaeger, 1973. Some pharmacologic and
toxicologic effects of di-2-ethylhexyl phthalate (DEHP) and
other plasticizerso Environ, Health Perspect. Jan. 53.
C-79
-------
Kubin, R.J., et al. 1979. Ames mutagenic assay of a series
of phthalic acid esters: positive response of the dimethyl
and diethyl esters in TA 100. Abstract. Soc. Toxicol. Annu.
Meet. New Orleans, March 11.
Schulz, C.O., and R.J. Rubin. 1973. Distribution, metabolism
and excretion of di-2-ethylhexyl phthalate in the rat.
Environ. Health Perspect. Jan. 123.
Seth, P.K., et al. Biochemical changes induced by di-2-
ethylhexyl phthalate in rat liver. Page 423 in Environmental
biology. Interprint Publications, New Delhi, India.
Shaffer, C.B., et al. 1945. Acute and subacute toxicity of
di(2- ethyhexyl) phthalate with note upon its metabolism.
Jour. Ind. Hyg. Toxicol. 27: 130.
Shibko, S.I. 1974. Toxicology of phthalic acid ester. In
Environmental quality and food supply.
Sidwell, S.I., et al. 1974. Composition of the edible por-
tion of raw (fresh or frozen) crustaceans, finfish, and mol-
lusks. I. Protein, fat, moisture, ash, carbohydrate, energy
value, and chlolesterol. Mar. Fish. Rev. 36: 21.
C-80
-------
Singh, A.R., et al. 1974. Mutagenic and antifertility sensi-
tivities of mice to di-2-ethylhexyl phthalate (DEHP) and
dimethoxyethyl phthalate (DMEP). Toxicol. Appl. Pharmacol.
29: 35.
Singh, A.R., et al. 1975. Maternal-fetal transfer of 14C-di-
2-ethylhexyl phthalate and ^-4C-diethyl phthalate in rats.
Jour. Pharm. Sci. 64: 1347.
Smith, C.C. 1953. Toxicity of butyl stearate, dibutyl
sebacate, dibutyl phthalate and methoxyethyl oleate. Arch.
Ind. Hyg. 7: 310.
Smith, O.M. 1924. Toxic properties of diethylphthalate.
Jour. Am. Pharm. Assoc. 13: 812.
Spasovski, M. 1964. The maximum allowable concentration of
dibutyl phthalate. (Translated title). Khigiena. 7: 38.
Srivastava, S.P., et al. 1975. Biochemical effects of di-2-
ethylhexyl phthalate. Environ. Physiol. Biochem. 5: 178.
Stern, I.J., et al. 1977. Physiochemical aspects of the ex-
traction in blood and the disposition in rats of di-(2-ethyl-
hexyl) phthalate plasticizer. Toxicol. Appl. Pharmacol. 41:
507.
C-81
-------
Swinyard, E.A., et al. 1976. Nonspecific effect of bis(2-
ethylhexyl) phthalate on hexobarbital sleep time. J. Phartn.
Sci. 65: 733.
Tanaka, A., et al. 1975. Biochemical studies on phthalic es-
ters. I. Elimination, distribution and metabolism of di(2-
ethylhexyl) phthalate in rats. Toxicology. 4: 253.
Thomas, J.A., et al. 1978. A review of the biological ef-
fects of di(-2-ethylhexyl) phthalate. Toxicol. Appl.
Pharmacol. 45: 1.
Tomita, I., et al. 1977. Phthalic acid esters in various
foodstuffs anmd biological materials. Ecotoxicology and
Environmental Safety. 1: 275.
Turner, J.H., et al. 1974. An evaluation of effects of
diethylhexyl phthalate (DEHP) on mitotically capable cells in
blood packs. Transfusion. 14: 560.
U.S. EPA. 1978. In-depth studies on health and environmental
impacts of selected water pollutants. U.S. Environ. Prot.
Agency, Contract No. 68-01-4646.
Veith, G.D., et al. An evaluation of using partition coeffi-
cients and water solubility to estimate bioconcentration
factors for organic chemicals in fish. (Manuscript).
C-82
-------
Vessman, J.,' and G. Rietz. 1978. Formation of mono(ethyl-
hexyl) phthalate from di(ethylhexyl) phthalate in human
plasma stored in PVC bags and its presence in fractionated
plasma proteins. Vox Sanguinis. 35: 75.
Waddell, W.M., et al. 1977. The distribution in mice of in-
travenously administered 14c-di-2-ethylhexyl phthalate deter-
mined by whole-body autoradiography. Toxicol. Appl.
Pharmacol. 39: 339.
Wallin, R.F., et al. 1974. Di(2-ethylhexyl) phthalate (DEHP)
metabolism in animals and post-transfusion tissue levels in
man. Bull. Parenteral Drug Assoc. 28: 278.
4
Walter, C.W. 1951. A technique for collection, storage, and
administration of unadulterated whole blood. In: Surgical
Forum 1950, American College of Surgeons, Philadelphia,
Saunders.
C-83
------- |