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    DBP.   Kozumbo et  al.  (1982) found  the  ortho dlester, DBF, to produce  a
dose-related mutagenlc response  1n  a modified  version  of the reverse  muta-
tion  pla :e  Incorporation  assay  In  Salmonella  (Ames  test).   This activity,
which wa-,  observed  only  In  strain  TA100,  a detector of  both base pair  and
frameshlft mutagens,  was  eliminated upon addition  of  S9.  In addition,  DBP
showed seme  evidence  of  clastogenU activity In Chinese  hamster  flbroblasts
(Ishldate and OdasMma, 1977) (see  Table V-10).

    PEP.    DEP  Is a direct-acting mutagen  for Salmonella  typhlmurlum  (Rubin
et al., 1979).   Seed  (1982)  found DEP  weakly  mutagenlc  1n a  forward  mutation
assay 1n Salmonelja typhlmurlum (see Table  V-10).

    PHP.    Extracted  urine  of  rats   administered  2 g/kg  DMP  l.p. was  found
not  to te  mutagenlc  (Kozumbo  et  al.,  1982).    In  vitro assays  by  these
authors showed that S9-assoc1ated esterases hydrolyzed  DMP to the monoester,
which has  not  been  shown  to be mutagenlc 1n  the Ames assay,  and  to  methanol
thereby  eliminating  Us  mutagenlc  capacity.   An  abstract by Yurchenko  and
Glelbermai  (1980)  Indicates that  DMP  Is  not positive  In a mouse  dominant
lethal test.   Rubin  et al.   (1979)  and  Kozumbo et  al. (1982)  found DMP  to be
a  direct-acting  mutagen  for Salmonella typhlmurlum.   In  a forward  mutation
assay DMP was  weakly  mutagenlc  In Salmonella  typh1mur1um  conducted  In  liquid
suspensloi  (Seed, 1982).   For more Information see Table V-10.

CarclnogeilcUy
    The  nost  conclusive   Information   on  the  carclnogenldty of  PAEs  was
obtained  From  bloassays performed by the NTP.   HUbourn and  Montesano  (1982)
reviewed  the  results  from  carclnogenldty  testing  of  PAEs conducted  prior to

04750                                V-86                            07/03/91
-------
the  NTP bloassays  and  concluded  that  all  the  studies were  limited  with
respect to study design or reporting,  making  the results  Inconclusive.

    DEHP.   Carc1nogen1c1ty  studies   have  been  conducted  by  the  NTP  for
several PAEs  Including  B8P  and  DEHP   (Kluwe et  a!.,  1982a,b;  Huff  and  Kluwe,
1984).   The  tested  PAEs  discussed  In  this   document  are  OEHP  and  BBP.
Essentially the  same  protocols  were  used  for  each compound.  The  compounds
(administered 1n  the  diet)  were tested for 2 years  1n both  Fischer  344 rats
and  B6C3F1  mice  using  an untreated  control  group, a  low-dose  group  and  a
                      t
high-dose group.   The high-dose used In  testing  was  the estimated  maximum
tolerated  dose   (MTD)   determined  by  preceedlng  90-day subchronk  feeding
studies prior to the  chronic exposure  bloassay.   The low-dose was  one-half
of  the estimated  MTD.    For  each  dose  group,   50  animals   of  each sex  and
species  were  tested.    Animals that died  during  the  study  and  animals
sacrificed at the end  of the study were subjected  to a  gross  necropsy and  a
complete mlcropathologlc  examination.  Statistical  comparisons  of  Incidences
of animals with  systemic  pathology, especially  tumors at specific  anatomical
sites  and  of survival  and  body weight  gain, were made using both  palrwlse
comparisons  (Fisher's  exact  test)  and  trend  tests (Cochran Armltage  trend
test).
    DEHP was administered  1n  the diet  for  103  weeks  at levels of 0, 6000 and
12,000 ppm  for  male and female F344 rats  and  0,  3000 and 6000  ppm for  male
and  female  B6C3F1  mice  (NTP,  1982a).    In  this  study procedure  rodent  meal
was  provided   In   such  a  way   that   measured   food  consumption  actually
represented  significant  spillage  and   waste  rather   than  true  food  Intake.
For  this  reason a  standard food consumption rate  of 13% of  mouse  and  5% of
04750
V-87
07/03/91
-------
rat body weight  was  used In the dose  conversion.   Corresponding dose levels
are 300  and  600 mg/kg/day for  rats  and  390 and 780 mg/kg/day  for  mice {low
and high dose,  respectively).   No clinical  signs  of  toxldty  were observed
In  either  rats  or  mice.  Survival  was not  significantly decreased  In  any
group  except  the female  mice  receiving the  low-dose  level.   The  Increased
number of  deaths In this group were not attributed to  OEHP administration,
since  pathologic  changes In tissues were  not observed and  excessive deaths
did not  occur  at the  higher  dose.    The  nonneoplasUc  lesions  observed  1n
this stuily were discussed previously under  chronic  toxldty.

    Spec f1c  Incidences  of  neoplastlc  lesions  for  the  various  treatment
groups are  presented In  Table V-ll.  The  major neoplastlc  effect observed
among  animals  treated with  OEHP was development of  liver  tumors.  The Inci-
dence  of animals with  hepatocellular  carcinomas was  significantly  Increased
among  female  rats fed  DEHP  at  either  300 or  600 mg/kg bw  when compared with
controls   Hale  rats  experienced  a significant  Increase of  the Incidence of
hepatoce"lular  carcinomas  and  neoplastlc  nodules  only  In  the 600 mg/kg  bw
DEHP  group.   Significant  dose-related  trends  for  Increased  numbers  of
animals  bearing  hepatocellular  carcinomas  and  for  Increased  numbers  of
animals  >ear1ng  either  hepatocellular  carcinomas or neoplastlc nodules were
found  1r  both  male  and  female  groups.   Among  male  rats,  Incidences  of
animals  with  pituitary  tumors, thyroid C-cell  tumors or  testlcular Inter-
stitial-cell  tumors  were all  significantly reduced among  treated  groups  by
both  pal-wise comparison and  trend  tests   (see  Table  V-ll).   Comparison  of
occurrence  of non-neoplastlc  effects  In  rats with  occurrence  of neoplastlc
effects  showed  that  the presence of testlcular  Interstitial  cell tumors was
n
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04750
V-89
07/03/91
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hypertrophy or  the  presence  of testlcular degeneration.   In  addition to the
tumors  presented  In  Table  V-11,  myelomonocytlc  leukemia,  mammary flbro-
adenoma,  clltoral   gland  carcinoma  and  uterine  endometrlal  stromal polyps
were observed  In one or  more  rats, but their  Incidences  1n  treated animals
did not dffer significantly from those Found 1n controls.

    The  lumber  of  mice  bearing hepatocellular carcinomas  was  significantly
Increasec  In  both  male  and female  groups  receiving  the high  dose (780 mg/kg
bw) of  D:HP and In  female mice receiving the  low dose  (390 mg/kg  bw)  (see
                     t
Table  V-ll).   Trend tests  showed  significant dose-related effects  for  both
sexes.  Metastases  of  the hepatocellular  tumors  to the  lungs  were  found In
12 male  and 8  female  DEHP  treated mice  bearing  hepatocellular  carcinomas.
Pulmonarj  metastases  were not  found 1n any  of  the control mice  with liver
tumors.   Incidences of  mice  with  hepatocellular  adenomas  did   not differ
significantly  from controls;  however,   the  Incidences  of  mice  with either
hepatocellular  carcinoma  or  adenoma was  significantly  Increased at both  dose
levels  IT  both sexes,  and  significant   dose-related  trends   were  present.
Lymphomas,  hemanglomas,   mammary   gland   adenocarclnomas  and   alveolar  or
bronchlolar carcinomas  or adenomas  were  also  found  In one or more treated
mice,  but  Incidences did not  differ  significantly from those  observed  1n
controls.   In conclusion,  the  DEHP  feeding studies  In  rats and  mice  Indicate
that   statistically  significant   Increases   1n  hepatocellular  carcinomas,
neoplastlc  nodules  and  adenomas occurred.   These tumors were  found  1n  both
species and both sexes.   There  were metastases of  hepatocellular  tumors  to
the lungj of treated mice.
04750                                V-90                            07/03/91
-------
     A  summary of the  Interpretation of  the  bloassay results by NTP 1s shown
 In  Table  V-12.   The only compound for  which there was clear evidence of car-
 diogenlclty  was OEHP.  In both  cases,  the  effect  observed was an Increased
 Incidence  of  liver  carcinomas  In  mice and rats  (Huff and Kluwe, 1984).
    Northrup  et al.  (1982) criticized  the conclusions  drawn from  the NTP
 DEHP  study (Kluwe et  al.,  1982a,b; Huff and  Kluwe,  1984)  claiming that the
 results could  not  be  Interpreted as showing a carcinogenic effect.  Northrup
 cited  as  one  problem  that the  designated  maximum tolerated  dose (MTO) had
 been  exceeded  (based  on  differences 1n  body  weight  gain) In  both rats and
 m1-:e  at  several  of the  treated groups.  Another  criticism of the study was
 that  there was  a  significantly lower  Incidence of  tumor  bearing  animals
 among  female  mice  used  1n  the control groups.   When  all  the control groups
 (both  rats and  mice) were pooled,  the  Incidence  of total   primary tumors
 associated  with  DEHP  treatment  was  no  different  for  all   control  groups
 except among  male  rats,  which  showed  a decrease  1n  total  number  of  tumors.
 Finally, Northrup  et  al.  (1982) claimed  that  critical  data on food consump-
 tion,  nutritional  status,  clinical   signs,  clinical  pathology and  Intestinal
 microorganisms  were lacking.   Northrup  et  al.  (1982)  also  felt that  the
 Incidence  of  liver tumors  could  have  been  Influenced by  the altered Intes-
 tinal  flora  Induced  by  DEHP.    The authors postulated  that   the  effects  of
 DEHP  were  attributable to  eplgenetlc  mechanisms of  cardnogen1c1ty  such  as
 chronic tissue  Injury,  nutritional   deficiency,  hormonal  Imbalance or promo-
 tional activity,  since  evidence of direct  genotoxlc effects  were lacking.
 Also,  Northrup  et  al.   (1982)  felt   that   because  DEHP  1s  metabolized
 differently  1n rats  than  In   humans,  effects  In  these  rodents   cannot  be
 extrapolated to Indicate  human risk.
0^750
V-91
07/03/91
-------
                                  TABLE V-12

       Summary of the Carcinogenic Effects of OEHP on the NTP Bloassays
                     and  Interpretation  of  These  Findings*
 Jest
Chemical
Species
Sex
Neoplasms
Interpretation15
  DEHP
 rats
         Liver  neoplastlc
         nodules/carcinomas
                     Some evidence
rats
mice
F
M&F
Liver carcinomas
Liver carcinomas
Clear evidence
Clear evidence
aSource: Huff and Kluwe, 1984

''Evidence  of  CardnogenlcUy--  Five  categories  of  Interpretative  conclu-
 sions  h.ive  been  adopted  for use  1n  the  NTP Technical  Reports  series  to
 specifically  emphasize  consistency  and  the concept of  actual evidence  of
 carclnocenHHy.  For each  definitive  study  result  (male  rats,  female  rats,
 male mice,  female mice)  one category 1s selected to describe  the  findings.
 This caiegory  refers  to the  strength  of  the experimental evidence  and  not
 to either potency or mechanism (Huff and Kluwe,  1984).
047SO
                       V-92
                                          07/03/91
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    On the other  hand,  Kluwe  et al. (1983) defended  the  conclusions  reached
in  the  NTP  study on DEHP  (KTuwe  et al.,  1982a)  by noting that the MTD was
estimated based  on  prechronlc oral studies,  and  that the MTD was not  tech-
nically exceeded  since  survival of animals was  not adversely affected.   In
response  to  the other  criticisms,  H  was noted  that  the Hver tumors were
Increased regardless of  which set  of  historical  control  data were used;  the
DEHP  bloassay  was  conducted   using  state-of-the-art  procedures  for  animal
cardnogenlclty  testing, and  that  the  results of the  bloassay were  approved
by  Independent  peer  review  panels.   Kluwe et al. (1983)  also noted  that  the
metabolic difference  between  rodents  and  humans,  cited  by  Northrup et  al.
(1982), would  not  be expected  to  affect the response  to the  hepatocarclno-
genlc  effects  of  DEHP  observed   In  rodents.  The authors  noted  that  the
International  Agency  for   Research  on  Cancer   (1ARC, 1982)  working  group
reviewed  the study  and  concluded  that  there was  "sufficient  evidence  for
cardnogenlclty  of  DEHP   1n   mice  and  rats".    There   Is   some   evidence
suggesting that  peroxlsome  proliferation, which occurs  In both  mice  and rats
at  the  dose  levels  used  1n  the  NTP  bloassay.   Is  Involved  1n a  secondary
mechanism of  cancer  Induction (Reddy et al., 1986).   Peroxlsomal  prolifera-
tion Is discussed In detail 1n Chapter  VII, Mechanisms of  Toxlclty.
    Similar  results  were  reported  In  cynomolgus  monkeys   (Short  et  al.,
1937).   In  this study  no treatment-related evidence of  hepatic  peroxlsomal
proliferation  was  found  In  monkeys  exposed to  levels  <500  mg/kg/day  DEHP.
Exposure to similar  levels  (100,  1000,  6000,  12000 and  25000 ppm) of DEHP In
rats  produced  hepatic  peroxlsomal  proliferation.   It  Is  difficult to compare
exposure  levels since monkeys  were  administered  bolus  doses and rats  were
administered   feed.   For  further  detail  see  the  metabolism  section  of
Chapter III.
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    Ward et al.  (1983)  studied the  patterns  of promotion  of  hepatocellular
neoplasla  by   OEHP   and  phenobarbHol   (PB)  following  Initiation  by  1.p.
dlethylnltrosamlne (OEN).   B6C3F1  mice were given a  single  l.p.  Injection  of
80 mg/kg of DEN  at  4 weeks of  age  followed  by  oral administration of  PB  or
DEHP  beginning 2 weeks  after  DEN  Injection and continuing  for <6  months.
DEHP  was administered  In  the  diet at concentrations of  3000,  6000  or 12.000
ppm,  and PB was  given  In drinking water  at  500  ppm.  Few fod  of hyperplasla
were  fourd  In  the liver  at 2, 4  or 6 months  In  animals  exposed  only  to DEN,
PB or DEHP,  while numerous fod  and hepatocellular neoplasms were  found  1n
                     /
mice  tree ted  with  DEHP  or PB  after Initiation  with  DEN.   The pattern  of
response  of  DEHP  differed   from  that   of  PB.    In DEHP-exposed  mice,  the
numbers  cf  foci  did not  Increase between 4 and  6 months as  they did  for PB,
but  the fod  did Increase  In  mean  diameter  and  volume  as  the  study pro-
gressed.   Fod and  tumors  appeared earlier 1n  the  higher  dose group  of DEHP
and,  alUough  the number  of  fod per unit  volume  of  liver was  similar for
all  DEHP dose groups,  the  volumes  of the foci  were dose-related.   The type
of  hepatccytes  found  In  the foci  and  neoplasms differed  for PB  and  DFHP;
those fot   PB  were  predominantly  eoslnophlllc  hepatocytes  while  those  In
DEHP-trealed  mice were predominantly basophlllc and were  more  malignant  In
appearanc;.   After  6 months  exposure,  the  neoplasms  1n the  high-dose DEHP
and  DEN n.lce were  significantly  larger  (p<0.02) than those  for  PB and DEN,
although hlstochemlstry  revealed  similarities In the  lesions.   DEHP  did not
exhibit  liltlatlng  action  when  given once orally followed  by PB for  6 months
 In drinking water.

     Ward et al.  (1986)  found  that  DEHP  did not  cause tumor  promotion  In
 female F3U/NCr  rat livers.   Rats were  Initially Injected  with 282 mg/kg DEN


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ami  thtm  fed diets  containing  12,000 ppm DEHP  or  placed on  drinking  water
containing  500  ppm PB.  Animals were  sacrificed after 14 days  of  exposure.
DEHP  failed to  Increase the  number  or  size of  focal  hepatocellular  prollf-
eratlve lesions  (FHPL).  The FHPL  were  morphologically similar between  DEN
and  DEN-DEHP  treated rats.   Based  on  the  above studies (Ward et al.,  1983,
1986)  the  Investigators   suggest   that  liver   cell  replication  1s  not  a
requirement  for  tumor  promotion and  that  the  hepatomegaly  Induced  by  DEHP
appears to be a  consequence of Increased  size of parenchymal  cells.

    Garvey  et  al.  (1987)  found that  a  single  oral  dose of  10 g/kg  or  12
weeks  of   feeding  1.2%  DEHP did  not serve to  Initiate  carclnogenesls  In
female  F344  rats.   Promoting  agents were 2-acetylamlnofluorene,   for  the
single  dose,  and  carbon  tetrachloMde  and P8  for  the  12-week study.   In
addition Williams  et al.  (1987)  demonstrated that  DEHP had  no Initiating or
enhancing effect on  male rat  carclnogenesls  when DEHP was  given  alone for 24
weeks  or   for  7  weeks  followed  by  the  PB.   The  absence  of   enhanced
development  of  foci In  DEHP-treated  rats   Is  also Indicative of  a  lack  of
prDmotlng activity (Williams et  al.. 1987).
    B6P.   BBP  was fed  for  ~2 years  to  both male  and  female rats and  male
and female mice at concentrations of  0,  6000  and  12,000 ppm (0,  780 and  1S60
mg/kg/day,  respectively)  (Kluwe  et   al..  1982b;  NTP,  1982b).   Body  weight
gains were decreased In  male  and female mice  and  In  female rats  Ingesting
BBP;  however,   survival   among  these  groups   was   not  affected.   Excessive
mortality  occurred among  male  rats  treated with 6000 and  12.000 ppm  BBP due
to apparent  Internal  hemorrhaglng,  after -3 months of  exposure.   Due to the
high  mortality the  study  of  male   rats  was  terminated  early,  precluding
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evaluatlcn  of  the  animals  for tumorlgenlc responses.   Incidences  of  tumors
at  specific anatomical  sites  In BBP-treated  male or  female mice  did not
differ  s  gnlflcantly from  controls.   However,  the  Incidence of mononuclear
cell  leukemia  was  greater  among  the  high-dose   female   rats  than  among
controls.   The  Incidences  of leukemia  In  female  rats  are presented In Table
V-13.   Although  the Increase  In  leukemia  was  statistically significant, the
biologic  relevance  of  this  finding  was questioned   due  to  considerable
variation 1n the  background Incidence of  mononuclear cell leukemia  In Fisher
344  rats.   The  conclusions  reached by  the peer  review  group of this  study
Indicated  that BBP "was  probably  carcinogenic  1n  F344  female   rats".   A
summary  cf  the Interpretive conclusions  drawn  from the  NTP carclnogenesls
testing o:  68P  Is shown 1n  Table  V-14.   BBP was  not carcinogenic  in mice of
either  sex.   In  reports of a  26-week subchronlc  study,  NTP  (1985,  1986)
revealed :Ignlflcantly  reduced  total  bone  marrow  cell  counts at  the 0.03 and
2.5%  dose   groups,  but  not at  the   0.09,  0.28  or  0.83X  dose  groups  when
compared vlth controls.  This  change  was  comprised  primarily of  decreases In
neutrophll  metamyelocytes,   bands,  segmenters,  lymphocytes,  and  basophlllc
rubrlcytes.

    The N" P 1s currently repeating   the  rat  portion of  the  cacner  bloassay
for  BBP.   Testing  began  1n Oune, 1989  (NTP,  1991).  When  Information from
this  study  becomes  available,  the welght-of-evldence for the carclnogenlclty
of BBP will  be re-evaluated.

    Using  the   results  of  the  NTP   cardnogenlsls  bloassays, Kluwe  (1986)
compared tie carcinogenic  effects of  OEHP  and  BBP  and  related  compounds  to
determine the  structure-activity relationships.   Among the  PAEs shown  to  be

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                                  TABLE  V-13
       Incidences  of  Female  Rats  with Tumors of  the HefKatopo1et1c System
                1n the NTP Cardnogen1c1ty Bloasssay  of BBPa>D

Hematopoletlc System Tumor

Myelomonocytlc leukemia
Lymphoma
Hyelomonocytlc leukemia or lymphoma


Control
7/49
0/49
7/49
Incidence

Low-Dose
7/49
0/49
7/49


High-Dose
18/50C
1/50
19/50C
aSource: Kluwe at al.. 1982b; NTP, 1982b

^Female Fischer  344  rats were  fed  diets  containing 0  (control),  6000  (low-
 dose), 12,000  ppm (high-dose)  of  BBP for -2  years.   The ratios of  female
 rats  bearing tumors  of the  hematopoletlc  system  to the  total  number  of
 female rats examined microscopically are  depicted.

cS1gn1fIcantly greater than  controls, p<0.05
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                                  TABLE  V-14

        Summary of  the Carcinogenic Effects of BBP In the NTP Bloassays
                    and  Interpretation of These Findings3
  Test
Chemical
Species
Sex
Neoplasms
Interpretat1onb
  BBP
 rats

 rats

 mice
 M

 F

M&F
                                           Leukemia
                  Inadequate study

                  Some evidence

                  No evidence
aSource: luff and Kluwe, 1984

^Evidence  of  Carclnogenlclty--  Five  categories  of  Interpretative  conclu-
 sions  have  been adopted  for use  1n the  NTP Technical  Reports  series  to
 spedflcjlly  emphasize  consistency and  the concept of  actual  evidence  of
 carclnogjnlcHy.  For each definitive study result  (male  rats,  female  rats.
 male mice,  female mice)  one  category Is selected to describe the  findings.
 This category  refers  to the  strength of  the  experimental evidence  and  not
 to elthe- potency or mechanism (Huff and Kluwe,  1984).
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potentially  carcinogenic,  the  target  sites of  carcinogenic action  varied.
For  example,  DEHP  Induced  hepatocellular  carcinoma,  while  8BP was  associated
with effects  of  the  hematopoletlc  system.   It was  concluded,  therefore,  that
the  carclnogenlclty  of PAEs may  not  be  due  to  the  acH1v1ty of  the phthalate
moiety but  rather  determined  by the moiety attached  to  the phthalate, or  to
a metabolic  byproduct.   Support  for  such an argument  1s given  by  studies  of
compounds   containing   the  2-ethylhexyl  moiety.     Comparison  of   results
obtained  for  DEHP,  DEHA and  two  other  compounds  [dt(2-ethylhexylJphosphate
and  2-ethylhexylsulfate]   containing  the 2-ethylhexyl  moiety  revealed  that
all  four  compounds  possessed   some  hepatocarclnogenlc  activity  In  female
mice.  The  related compounds  will  not be discussed  In  this document.   Thus,
these results  may  Indicate that compounds containing  the  2-ethylhexyl  group
may have a propensity for causing hepatocardnogenlclty 1n  female mice.

    DSP.   Data  regarding  the  carclnogenUHy  of  OBP could  not  be  located
1n the available literature.

    DE_P.   Data  regarding  the  carclnogenlclty  of  DEP could  not  be  located
In the available literature.

    PHP.   Data  regarding  the  carclnogenlclty  of  DMP could  not  be  located
In the available literature.
Summary
    The acute  toxUHy  of  PAEs  tends to be  Inversely  related  to  the  molecu-
lar weight  of  the compound.  Signs  of  long-term toxlclty  Include  decreased
body  weight  gain  and  Increased  liver,  and  In  some  cases, kidney  weights.
Target organs of PAEs,  particularly DEHP,  are the testes,  liver and  kidney.
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    The fepatotoxH  effects  of PAEs  have  been studied by  numerous  Investi-
gators 1n a variety  of  species.   Seth (1982)  reviewed the hepatic effects of
PAEs  anc   described  both  the  morphologic   and   biochemical   alterations
attrlbutatjle  to PAE  exposure.   Host  Investigators  have  used  DEHP as  the
representative PAE 1n testing.  Generally, enlargement  of  the  Hver  has been
observed  following  oral  or  l.p.  administration  of  PAEs.   Examination  of
tissue  fi om enlarged  mouse,  rat,  hamster and  monkey  livers  has  revealed
changes 1n  morphology  and biochemical constituents.   Oral  administration of
OEHP  for  21 days  was  reported to  cause dilation  of smooth and  rough  endo-
plasmlc retlculum, mltochondrlal  swelling  and  Increase.1n  mlcrobodles  In rat
Hver (La
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    TesUcular Injury Induced by PAEs appears  to  be  species  specific  to  some
extent.  The  rat,  mouse,  guinea  pig and ferret were  susceptible  to  testlcu-
lar  Injury  from  OEHP and DSP while  the hamster  appeared to be resistant  to
the  gonadal  effects  of  these compounds  and  the  corresponding monoesters  at
the dose levels and durat'ons tested (GangolU, 1982}.
    Studies on  the embryotoxUHy of  PAEs seem  consistent  with other  data
obtained  using   different   toxlcologlc  endpolnts  {Tyl,  1988;  NTP  1984a,b,
1985; Mitchell  et a!.,  1985; Dostal  et  al.,  1987a),  that  1s,  there  1s  a
range of  toxlcltles  that varies as a  function  of the PAE being  tested  and,
1n -general, high concentrations  of this  chemical  are required  to  produce  a
teratogenlc response.   Most studies used  the mouse or  the  rat as  the  test
subject  and 1n those situations wherein  a  teratogenlc response  occurred,  the
target was generally the skeletal  system  (Shlota  and  N1sh1mura,  1982;  Tomlta
et al.,  1982a;  Singh et al.,  1972).   Based on  the  high doses  used and  the
differences 1n  PAE  metabolism between  man and  these  test  species,  It  1s
difficult at this time  to define clearly the risk for  the human  population.

    PAEs are generally  regarded as nonmutagenlc  although mutagenlc  responses
have been shown  for  some PAEs  1n  some  tests.   DEHP  1s apparently metabolized
to a nongenotoxlc form 1n Intact animals but not  by  tissue  preparations.
    DEHP and BBP have been  tested  for  cardnogenlclty  In  2-year  NCI/NTP  car-
c1f>ogen1c1ty bloassays.   DEHP  was  found to  Induce  hepatocellular  carcinomas
1n both rats and  mice.   Increased  mononuclear cell leukemia was observed  in
female rats exposed  to  BBP.  Data  regarding the carclnogenlclty of  DBP.  DEP
and DMP could not be located 1n the available literature.
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                        VI.  HEALTH EFFECTS  IN HUMANS
Introduction
    Althojgh PAEs are considered  to  have a  low order  of  toxUHy,  much  con-
cern has been  generated by  the discovery that PAEs, such as  DEHP,  may  leach
from the plastic  tubing and  plastic  bags used for  blood  storage  (Jaeger  and
Rubin,  1970, 1972,  1973;  Peck  et  al., 1979). Studies  on  the  effects  of  PAEs
In  humans  have  largely  provided  Information about the pharmacoklnetlcs  of
the compounds.   Associations  between  exposure to  PAEs and  toxic effects  In
humans  han-e been  limited  by  the Inability to discern  doses and  responses  In
light  of   the  ubiquity of  phthalates  In the  environment.   DEHP  has  been
detected in both  transfused and nontransfused patients (Wallln et al.,  1974;
Rubin and  Nalr,  1973;  Jaeger  and  Rubin,  1972).  In addition  children may  be
exposed  to DEHP  In  products   such  as  pacifiers,   teethers.  squeeze  toys,
plastic b.iby pants and vinyl fabrics  covering playpen  pads.   A report by the
Consumer deducts Safety  Commission estimated possible Increased  cancer  risk
to  children exposed  to   the  above products  (CPSC, 1983).   The widespread
presence of PAEs  1n air,  water, food and  stored blood Indicates  that humans
are subject to environmental  and Industrial exposures to PAEs.

Clinical 
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    Shaffer  et  al.  (1945)  also  examined  the effects  of  DEHP after  dermal
exposures to the plastldzer.   Undiluted DEHP was  applied  to  the  backs of  23
human subjects as  patch  tests.   The compound was  left  1n  contact  for  7  days
and then reappHed on  the same  spots  after  10 days.   These exposures  did not
result  In any  type of erythema  or  other effects, suggesting  the  Irritating
and sensitizing potentials of DEHP are minimal.
    Jacobson et  al.  (1974) examined  the  effects  of DEHP on  tissue  cultures
of  ligman  dlplold  flbroblasts  established  from  skin  biopsies.   DEHP  was
solublllzed 1n  sera  collected and  stored  1n  polyvlnyl  chloride  {PVC}  blood
packs  under  standard  blood  bank  conditions   (4°C).   Tissue culture  medium
containing  15%  of   the  plastic  stored  serum was  used for  Incubation  of
cells.   The  degree  of  growth  Inhibition  of   the  human dlplold  flbroblasts
Increased with  DEHP  concentration.   At Incubation concentrations of  0.10 mM
and  0.18  mM OEHP, cell  growth was  Inhibited  by  20%  and 50%.  respectively.
These  DEHP levels  were  comparable  with  concentrations detected  1n  whole
blood stored 1n  PVC  blood packs at  4°C for 14  and  26  days,  respectively.  A
70%  Inhibition of cell growth  was  observed  when 0.70 mM DEHP was used, which
was  the concentration detected  1n  platelet  concentrations stored  at  22°C for
48 hours.
    Chromosomal effects  of  DEHP (Stenchever et al.,  1976)  were Investigated
on human leukocytes from  the  blood  of  two  male and two female healthy donors
1n their early twenties and  on  fetal  lung cells  established  from  a 16-week
fetus  delivered  by  hysterotomy.   DEHP  was   solublllzed  In  Polysorbate  80
(Tween)  (1:3,   vol:vol)  and  dispersed  1n  fetal   calf  serum  by  sonlcatlon.
AHquots were  diluted  to  final concentrations  of 0.06,  0.6, 6.0  and  60.0
jig  OEHP/mt  of  blood  for  the  leukocyte  Incubations.   Incubations  with
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DEHP  were  for 4  hours at  37°C.   Phytohemaglutlnln  was  then  added  for  30
minutes  t<>  Initiate  cell  division  and cells  were cultured  for 72  hours.
Mitosis was  Inhibited  by  addition  of  democoldne 2 hours  prior  to  harvest-
Ing.  Metaphase  spreads were scored blindly for  chromosome abnormalities  on
Glemsa-stalned slides  prepared  from  these cultures.   Pooled  data  from  the
four  donors showed  no  statistical  differences  In chromosome breaks,  gaps  or
abnormal forms  at any  of  the  Incubation  concentrations when compared  with
control  ciltures.   Fetal   lung  cells  were Incubated  with 6.0  vq  DEHP  (In
Polysorbat?  80)/mi  medium   for   5 days.   No   significant   difference   1n
aneuploldy between study and control cultures was seen.
    Ishlka^a  et  al.  (1983)  determined that  platelet  function decreased  as
DEHP  concentrations  Increased In  PVC  blood  storage bags.   Platelets  demon-
strated  a  decrease  In ADP-lnduced  aggregation after  at least  2 hours  of
exposure  1o  100,  300  or  500  yg  DEHP/ma.   Maximum  aggregation  gradually
decreased  with  Incubation  time,  depending  on  the   DEHP  dose.    Platelets
renewed wl :h  fresh plasma showed a  restoration  of  aggregablHty.   The  degree
of restora:1on was decreased  with Increasing  DEHP dose.
    The ef:ects of DEHP  on  cultures  of  the  human  dlplold cell  strain,  WI-38,
were Investigated  by  Jones  et al. (1975).  Cultures  treated with  51,  69 and
160  yM DEHP  (soluble  concentrations  In  Incubation  media)   showed  a  sta-
tistically significant  decrease  In cell protein and  longer generation  times
when   compired  with  control  cultures.    As   Indicated  by   figures   these
decreases  were  dose  dependent.    Cells  treated  with  160  uM  DEHP  were  no
longer  viable  at  day 9  and  exhibited decreased  cell  density  on day  6  of
treatment.  The  dose, which  caused  50%  growth  Inhibition  (IDrg), was  cal-
culated to  be  70  iiM.   These  toxic effects  were greater  In replicating  cell
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populations  than  In  those  treated  after  reaching confluency.   Cells grown 1n
160  yM for  3  days and  subsequently  subcultured Into  control  medium showed
only 60% of  control growth after 5 days 1n control medium.
    BBP.   Malette and  von Haam  (1952)  Investigated  the  dermal  effects  of
various  phthalates.   A  100%  solution of  BBP  had moderately  Irritating  and
slightly  sensitizing  effects  when applied to white  rabbits  (methodology  not
specified).  Patch tests were  performed  on 15-30  human subjects and sensltl-
zatlon  tests  2  weeks  after   primary   Irritation   tests.   A  10%  solution
(vehicle  not defined), of  BBP was  applied to  the  human subjects.   A light
reaction  (not  described)  was  seen  In 12% of  those  tested.   The Irritative
effect was classified as moderate and no sensitizing effect was reported.

    DBF.   Atmospheric   exposures  to   DBP   were   studied   by  Hen'shlkova
(1971).   A  human olfactory  threshold   was  found  to  range  from  0.26-1.47
mg/m3.   Abnormal  encephalographlc  responses were  noted  In  three  subjects
at  atmospheric  DBP  levels of  0.12  and  0.15  mg/m3.   At  0.093  mg  DBP/m3,
conditioned  reflexes  were  not  observed.   A maximum  atmospheric  concentration
of 0.1 mg DBP/m3 was recommended.

    A single case of  accidental  1ngest1on  of  DBP  by  a 23-year-old adult male
has  been  reported  (Lefaux,  1968).   The  Individual  mistakenly  Ingested  a
spoonful  (-10 g)  of  DBP Instead of a laxative.   The Individual was hospita-
lized  the next  day  with  complaints  of  nausea  and  vertigo.   The  subject
exhibited signs of keratltls and  toxic nephritis  (excess albumen and  red  and
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white corpuscles  1n  the  urine).   Unspecified  treatment  Initiated Immediately
allowed Ue  subject  to leave  the  hospital  after 2 weeks without  any  after-
effects.

Epldemlolcqlc Studies
    MUkov  et  al.  (1973)  performed  a  cross-sectional  Investigation  of
workers e;:posed  to  phthalate  plastldzers  In the manufacture  of  artificial
leather and  PVC-based films.   The phthalates In use  Included  predominantly
DBP  and higher alkyl  phthalates   (DAP-789),  but  periodically DEHP and  BBP.
Some  formulations  contained  small  amounts  of  the sebacates  [dlbutyl sebacate
(DBS)  and dloctyl  sebacate  (DOS)]  or  adlpates  [dlbutyl  adlpate  (DBA)  and
dlocytl aclpate  (DOA)].   TMcresyl  phosphate (TCP) was  a  component  of the
Incombustible  materials  produced  1n  10-20% of machines  assigned  to  various
workers.   The  presence of these other agents  without  any attempt to account
for  confounding Is a major criticism of this  study.

     The study  population  consisted of  147  persons,  87  women and 60 men.  The
majority  |75%) of the population  was  <40  years  of  age  (mean  and  range not
given).  Exposure duration was divided Into three categories:  0.5-5.0 years
for  54, 6-10 years for 28 and 10-19 years  for 65 workers, respectively  (mean
and  range not  specified).   Job categories  Included:   60 primers, 28 calender
and  mill   )perators,  35  mixing apparatus  and paint millers, and 24 winders
and  final product  Inspectors.   A control population was not Identified.

     Ambient  exposure levels  to vapors  or aerosols of the plastlclzers (mixed
esters)  11  the  working  zone  of  the  primers  ranged  from  10-66  mg/m3.
 04760                                VI-5                             08/15/88
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Similar results were  reported  for  the work station  of  the  mm  and  calender
operators.   The  plastldzer  level   in  the mixture  preparation  section  was
found  to  be  1.7-40   mg/m3.    Other  contaminants   (vinyl  chloride,  carbon
monoxide and hydrochloric acid) around the  calenders  and  rollers were  either
below their maximum allowable concentrations or not detected.
    "he  test  procedures  Included  algeslmetry,   olfactometry,   audlometry,
vibration  sensitivity  and  vestlbular  function  by the  caloric  method  with
cold water  (60  ml at  19°C  for  20 seconds).  Clinical and  biochemical  blood
studies {sedimentation -rate and blllrubln level) were  also performed.

    The most  frequently  cited complaint was  of pain  In  the  upper  and  lower
extremities  accompanied  by  numbness  and spasms,  reported  1n  51.7% of  the
subjects with a  length of service 6-10 years and  In  81.6%  In those with >10
years,   PolyneurlUs was  found In 47  persons,  32 with  an  autonomlc-sensory
and  15 with  a  mixed  form.  The  Incidence  of  polyneurltls Increased  with
length  of  service.   In  3.4% of  the cases,  organic  disease  of  a  nonoccupa-
tlonal   character  was  noted  In  the  nervous system.   An  elevation  1n  the
threshold  for  sensitivity to  pain  was noted In  66.7%  of  the subjects,  and
sensitivity  to  vibration  was lowered  to  some  extent   1n  33.8%.   A marked
depression  In  vibration  sensitivity  was seen  only   In  those subjects  also
manifesting  a  significant  depression  of pain  sensitivity.  Of  81  subjects
undergoing  vestlbular  receptor  Investigations,  78%  were  found  to  have  a
depression of  the vestlbulosomatlc  reactions (absence or  lowering  of exdt-
abl'Hy).  This  depression  began  with the  first  years  of  this  occupational
contact, often In the  absence  of any health status complaints.   The majority
of  subjects  showed  an elevated  threshold   of  excitability  when   tested  by
047 (JO
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olfactometry,  especially  for  thymol  (82.1%)  but  also  for  camphor  and  tar
(50%) and less for rosemary  (33.4%).  This  elevation  Increased with duration
of servlct .   Audlometry  did not  reveal  any pathology  1n  auditory  sensitiv-
ity.   Blocd  studies  revealed a tendency to  slight  lowering  of the  number  of
platelets and  leukocytes,  hemoglobin  level  and blood color  Index.   A slight
retlculocytosls and a  tendency  to  acceleration of  the erythrocyte sedimenta-
tion rate among  the  female subjects (statistically significant,  but  p value
not stated)  was also noted.
                      t
    Thless  et al.  (1978a)  performed a  morbidity  study  on  101  workers  (97
males,  4 females) employed  1n  a  DEHP  production plant.   The  age range of the
workers  w.is  from 22-60  years  (no  mean  was   given  but  the  majority  were
between 35  and  55 years  of age).  Duration  of exposure was  between 4 months
and 35 years, with an  average  of  12  years.   In 1966 the plant  changed from a
batch process to  a continuous  process production  so that exposure to workers
was  reduced to only  the  processes of  removing samples  In  control  passages
and  during   decantatlon.    The  Investigators   concluded   that  the  negative
results reported  may  be  attributed to  lower exposures after  the processing
change  In   1966.   Samples   of  current  exposure concentrations  In  the  work
areas  In  question ranged  between  Q.0006 and  0.01  ppm  (detection  limit  not
stated).  A clinical  and  occupational   history was  taken  and the  clinical
examlnatloi  Included  vital  statistics,  EKG,   lung  X-ray,  and  a  complete
urinary status with  uric  add  and creatlnlne  clearance.   The  blood analyses
Included  <   differential  count  and   sedimentation  rate,  and  thymol,  total
protein,  >GOT,  SGPT,  
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    All  the  results of  the  examinations were  compared with  those  obtained
from  two  In-house  control  groups  with  possible  exposure  to  styrene  and
dlmethylcarbamlc  acid  chloride  (DMCC).   No significant differences  between
the  study  group and control  groups were found.   Even when  the  study  group
was  divided  by  age and time of  exposure  (greater and less  than  12  years  to
presumably account  for  the  change  1n production  process),  no  significant
differences were  seen.   Workers with  duration  of exposure  >20  years  (n*6),
with an  average  exposure  of  26.3  years, were  given  a neurologic examination
that Included:  tests of  the cerebral  nerves;  reflexes of  the arms,  legs and
abdominal skin; and sensitivity  to depth, pain  and vibration.  No neurologic
disease  or  toxic   nerve  damage  was  Indicated.    Analyses  of  absenteeism,
accident   rate   and   of  a   questionnaire   regarding   premature   births,
miscarriages   and  malformations were  also  negative.   Although the  study was
comprehensive In scope, 1t lacked  exposure  data prior to the conversion to a
continuous process  production  so  that a  definitive  conclusion  can not  be
ascertained.   As such,  however.  It represents  only  one  of two epidemlologic
studies  reported  to  date  on   subjects  with  DEHP   or  a  specific  phthalate
exposure.
    Thless et al.  (1978b)  also  reported  a  mortality study on these DEHP pro-
duction workers.   The study was  a  prospective cohort  survey  of  221  workers
con-pared  with  the general  population.   The  study  considered data prior  to
1916.   The  population was  derived  from 28  workers who had worked prior  to
1955, 85  workers  who had started between  1940-1965,  135  workers  who  started
after  1965  and  109  workers employed  at the  time  of  the  study.   Selection
criteria  for  Inclusion  In  the  study   population  were  not provided.   The
average observation  period  was  11.5  years.   Half  of the expected  deaths were
observed  1n  the  exposed   population.   Eight  cases  of  death were  due  to
04760
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cancer.  Tiless et al.  (1978b)  reported  one  case of bladder papllloma, which
was   significantly   different  from   that   expected.   However,   this   was
attributed to a single  case  and was  not  considered to represent an Increased
health risk.  Analysis  of  natural death cases,  after  minimal  observation of
5-10  years,  on  workers exposed to durations  of  5, 10, 15  or  >15  years,  did
not reveal an Increase 1n mortality with exposure duration.

    Thless  and  Flelg  (1979) also  performed  chromosomal  analysis on  blood
lymphocyte*  from  a  subset of  this  same  study population.   Lymphocytes  were
cultured from 10  exposed production workers  according -to  a modified  method
of  Hoorhead  et  al.  (1960).   The workers'  duration of exposure ranged  from
10-34  years  (mean =  22.1  years).  Lymphocytes  from 20 age-matched  workers
served as  controls.   It was  not  mentioned  whether  these controls  were  also
exposed to styrene and  DMCC  as  mentioned  previously.  One  hundred  metaphases
were  scored  for abnormalities on  lymphocytes  from each worker.  The specific
structural  abnormalities  were  not  defined,  but  were  categorized with  and
without gaps.   Neither  category appeared to  be  different  from  the controls
although statistical analyses were not stated.

    In  addition  to   Individuals  who  are occupatlonally   exposed,  research
Indicates  that  persons who  undergo blood  transfusions or  hemodlalysls  may
receive extensive amounts  of PAEs  (over  background Intakes)  as a  result of
the   leaching  of  compounds,  such  as  I1p1d  soluble  OEHP,   from  plastic
containers  or  catheter  tubing (Marcel  and  Noel,   1970;  Jaeger  and  Rubin,
1970. 1972,  1973).
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    Hlllman et  al.  (1975)  studied DEHP levels 1n  neonatal  heart  and  GI  tis-
sues.  The study  tissues were  obtained  from  three  Infants  who previously had
umbT.lcal catheters  In  place  but  had never  received blood  products and  from
14  Infants who  previously  had umbilical catheters and varying  quantities  of
blood products.   Control tissues  were  obtained from  eight  stillborn  Infants,
two  Hveborn  Infants  who  had  died without administration  of  any  blood  prod-
ucts  or   Insertion  of  catheters,  and  three older   subjects  who  had  not
received blood  products.  The  maximal  amount  of  DEHP  that  could have  leached
Into  the  blood  was determined  to be 13.9+fl.l mg  of  DEHP/5  cm of  catheter,
based upon  the mean  i'S.E.  of  the extraction  of four'No.  5 French  cath-
eters.  The maximal  amount  contributed by blood products  was estimated  at 4
vg/mi,  based   upon  a  reference  to Marcel   (1973).   The  potential   dosage
thus  ranged  from  0.04  mg  In   Infants  receiving  only  10  ml of blood  to 1.4
mg  in those  receiving  double exchange  (460 ml).    The  minimum  detection
limit for  DEHP was  -0.02  yg/g  of  tissue  under  the  conditions used.   No
correlation could be  made  between hours of  catheterlzatlon and DEHP  levels.
In general, the OEHP  levels In heart tissue  reflected  the  combined  dosage of
the numbers of  catheters and  amount  of  blood products  1n Infants  who  died In
<24  hours.   In Infants  who  lived  longer,   levels were  generally lower  and
less  correlated  with  dosage,  suggesting  that   some  blotransformatlon  and
clearance  of  DEHP  was  taking place.   The  mean  levels   of  DEHP for  heart
residue and  pressed extract  of the study  tissues,  1.27*0.42  and  0.66+0.22
ng/g,  respectively,  were   significantly  higher  than   the  corresponding
control  levels, <0.07+.0.03  and <0.07*0.04 yg/g.   Three  Infants who  died  of
necrotlzlng   enterocolHIs   and  who   previously  had  arterial   umbilical
catheters  In  place and  removed,  had  gut  residue levels  of  0.47,  0.63  and
0.16  yg/g.   These  levels were significantly higher  (p<0.05) than  those  1n
GI  tissues  from  Infants  without this disease.   These  higher DEHP  levels
04760
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may represent  Increased  uptake  or  decreased metabolism by  a  dying bowel.   A
direct causative  link  could not be  determined between exposure  to  DEHP and
the  deve'opment  of  necrotlzlng enterocoHUs.   However,  the study  demon-
strated  that   DEHP  accumulated   1n  the  tissues  of  critically 111  Infants.
Component; of  the catheters,  Including DEHP,  should  be  further  Investigated
as  poten .lal   vascular  or  GI   toxins,  according  to the  authors  of  this
Investlga ;1on.

    Another group that could be at high  risk for  the development  of  toxic
responses  to  exposure' to PAEs  may  be Individuals who undergo hemodlalysls.
Gibson  et al.  (1976)   studied  blood  samples  from  nine  patients  requiring
maintenance  hemodlalysls  both   before,  during  and  after  the  hemodlalysls
process  tj quantify the  levels of  DEHP  received by these  Individuals  from
blood  transfusion bags  and/or plastic  hemodlalysls  tubing.   Hemodlalysls was
performed  using reclrculatlng  single-pass  machines  and   colls.   Samples for
DEHP  analyses  were obtained at  15  and 30  minutes,  1,  2, 3,  4 and  5  hours,
and  1mmed ately after  dialysis  unless dialysis was  terminated earlier.   The
metabolic  fate and  toxldty of  these DEHP  levels  were not determined.   Esti-
mates  of  the   total amount  of  DEHP delivered  to  a  patient  during hemodlaly-
sls  rangeil from  1.5-150  mg for dlalyses  that lasted from 15 minutes  to  5
hours.

    Neergaard  et al.  (1971)  reported that  exposures  to DEHP may have  been
assodatec  with  the development  of abnormal  liver  function  tests  1n  three
patients  (two  men,  aged 25  and  40  years, and  one 25-year-old woman), follow-
ing  the  u: e of a  new  set  of  PVC blood tubings In hemodlalysls.   IR-analyses
of  DEHP  In  salt  solution  perfusates through  this  blood  tubing  set  ranged
from  10-2) mg/l.  UV-determ1nat1ons  using  the  perfusate  directly gave  a
04760                                VI-11                           08/15/88
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somewhat  higher  range,  20-50  mg/l.    DEHP  could   not  be  washed  out  by
perfuslon  of  three  other  commercially  available  blood  tubing.   Over  a  5
month  period  dialysis  machines  with  this  tubing  were  used  93  times.   Of
these  dlalyses,  75  were  performed  upon  the three  patients  who  developed
symptoms.   Symptoms  presented  by the  three  patients  after 10-15  dlalyses
(estimated dose of DEHP  not  given)  Included malaise,  fever,  abdominal pains,
nausea,  abnormal  serum  enzyme  levels  (LDH  and  SGOT),   Increased  serum
blllrubln,  and  1n  one  case,   jaundice.    Liver  biopsies  In  one  patient
revealed   changes    1n   accordance  with   so-called   nonspecific   reactive
hepatitis; and 1n  another  patient,  a hlstologlc  picture compatible  with  a
diagnosis of  viral  hepatitis.   Upon removal  from the  new  dialysis  machines
to  dialysis  systems  1n  which OEHP was  not detected, the conditions  of the
three  patients Improved.   A patient who  was returned  to  the  new  dialysis
machine  developed  a  more  severe relapse of  the symptomology until  she was
removed  to a different  dialysis  system.   Evidence as  to the  exact  etiology
of  Illness associated with  the  use of  the  new dialysis  machines  could not be
determined.
    In a recent study  (Pollack  et  al.,  1985b),  circulating concentrations of
OtHP   and   Its   desterlfled   phthallc   acid   products,   mono{0-ethylhexyl)
phthalate (HEHP)  and phthallc acid, were  quantltated  (HPLC/UV  monitor)  1n 11
patients.   These  patients  were  undergoing  maintenance  hemodlalysis  for
treatment of  renal  failure.   The patients  underwent hemodlalysis  3  times/
week,  4  hours/session,  and had  been receiving  treatment  ranging  from  1 week
to  12  years.   The mean  estimate  of  DEHP extracted during  a  single dialysis
session  for  the 11 patients was  105  mg (range 23.8-360  mg).   Serum choles-
terol, trlglycerldes and AAG concentrations  were  measured to  determine  their
04760
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Influence on  the  extraction  of DEHP Into blood.   Circulating  levels  of  DEHP
and  MEHP   (1.9U2.11   w/ml   and   1.33^0.58  vg/mi,   respectively)   during
dialysis  I1d  not correlate  wHh the  length  of  time  the  patients had  been
undergoing dialysis.  This,  together wHh the  observation  that blood  concen-
trations (if  OEHP  during 1nterd1alys1s were  similar  to those  1n  nondlalyzed
patients Indicate that  these compounds are effectively  removed from the  cir-
culation between  dialysis  sessions.   This  could represent metabolic  trans-
formation or  sequestration  of  UpophHU  phthalate  esters  Into  fatty  tis-
sues.  Thi'te  was  a  strong correlation,  however,  between  phthallc acid  con-
                      /
centratlons   (5.22*3.94  vq/ml)  and  the  length  In  years of  previous  dialy-
sis treatments  (rs+0.920,  p<0.001).  There also  was  no  apparent relationship
between the concentrations of OEHP  and either  of Us  metabolites,  Indicating
the need  for future metabolic  fate and pharmacoklnetlc Investigations.   Of
the biochemical factors  examined,  the  sum  of the  serum cholesterol and  trl-
glycerlde concentrations  correlated most  closely  wHh  the  Teachability  of
OEHP, altliough  the  association  was  weak (r=+0.565,  p~0.1).    Thus, although
hemodlalysls  patients  are exposed  to circulating DEHP,  the   consequence  of
long-term systemic exposures to  the ester  and Us metabolites  remains to  be
elucidated.
High Risk Subpopulatlons
    Although toxic effects of PAE exposure have not  been  conclusively  demon-
strated, 'ndlvlduals who  receive  exposures above  background  or  environmental
levels, si.ch  as  those requiring  hemodlalysls  or  blood transfusions, may  be
at higher risk  for  the development  of adverse reactions  to  these  compounds.
Parenteral  administration of  PAEs to  these Individuals may  prove  to be  more
toxic because these patients  are  critically 111 or subject to  differences  In
their ability to absorb, metabolize  and excrete the compounds.
04760
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Summary
    DIspHe  widespread  occurrence   of   PAEs,   Information   concerning   the
effects  of  human  exposure  1s  limited.   In one  study  of  acute  exposure,
administration of  5  or  10  g DEHP to  two adult males did  not  result  In  toxic
effects other  than mild  GI  disturbances.   Accidental Ingestlon of  10 g  DBF,
however, proved  more toxic  and caused  nausea,  vertigo, keratltls and  toxic
nephritis.   Dermal application of BBP on  humans  did  result  In Irritative but
not sensitizing  effects; however,  the application  of DEHP did  not  result  In
either effect.   Studies  on  human  tissue and cell cultures have demonstrated
Inhibition  of  cellular  growth  and decreases In  platelet  function.   However,
chromosomal effects did not  occur  1n human leukocytes and  fetal  lung cells.
    In  epldemlologlc  studies  the results  have been  largely confounded  by
exposure  to  multiple  chemicals  and  lack  of  quantitative  Information  on
levels  and  duration of  exposure.  One  group  of  Investigators  conducted  a
morbidity  study  on  101  workers  employed  In  a  DEHP  production  plant.
Clinical examination and blood analyses revealed  no  significant  differences
between the study  group  and  control  groups.  No neurologic disease  or  toxic
ne've damage  was  Indicated.   Although  the  study  was comprehensive  1n  scope
1t  lacked  exposure data prior  to a  process conversion  In  the plant.   In  a
cross-sectional  Investigation  of  147  persons  exposed   to  a  combination  of
phthalate plastldzers,  the  Incidence  of polyneurltls Increased with  length
of  service.   HematologU  studies  revealed  a  lowering of  the  number  of
platelets and  leukocytes,  hemoglobin  level and  blood color Index.   Exposure
to multiple agents make  It difficult to  Interpret  these  results.   Finally  In
a  prospective cohort  study  of  221  workers  exposed to-DEHP,  half of  the
expected deaths were observed  In  the exposed  population.   However,  selection
criteria for Inclusion  In the study population were not  provided.
04760
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    Parenteral  administration  of  PAEs  may   Involve  the  greatest  risk  for
toxic  effjcts,  especially  1n Individuals  requiring  blood  transfusions  or
hemodlalysls.   Despite  the fact  that PAEs  may leach  Into  the  contents  of
plastic b ood  bags or  plastic  tubes, reports  of  hepatitis In  hemodlalysls
patients 
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                         VII.   MECHANISMS OF  TOXICITY
Introduction
    The  relationship  between the  toxlcoklnetks  and  toxic  effects  of  PAEs
(and  their  metabolites)   has  not  been  fully  elucidated  because   of   the
relatively  expedient  clearance  and  minimal  tissue  accumulation  of these
compounds.  However,  Investigation  of  the  mechanisms  of  phthalate  toxlclty
has been  promoted  In part  by Interest  In  their  carcinogenic potential  and
their  widespread use and environmental disposition.
Interactions
    Concern  over  the  ability  of  PAEs   to  alter   biologic  responses   to
pharmacologlc agents  and  xenoblotlcs has  stimulated  research Into  the  pos-
sibility that  the blotransformatlon of  these chemicals  may  be modified  by
the acid esters.  DEHP and D8P  have  been  found  to Interact with  the toxicHy
of  other  compounds   1n  a   synerglstlc   or  antagonistic manner.    Carbon
tetrachlorlde  was  found   to   act  synerglstlcally  with   DEHP  by  producing
extensive  necrosis  of parenchymal cells  1n  rat  liver  (Seth  et a!.,  1979}.
DEHP  significantly  (p<0.05)   Increased  barbiturate-Induced sleeping time  In
male  mice  {Rubin and  Jaeger,  1973).   A   synerglstlc  effect  was noted  when
DEHP   or   DBP   (applied   prior   to   the  application   of   organophosphate
Insecticides)  Increased  the  mortality of  female  house flies.   When applied
simultaneously,   DEHP   or   DBP  reduced   the   toxUHy   of   organophosphate
Insecticides  to house  flies  (Al-Badry and  Knowles,  1980).  Antagonism  was
noted  between  the effects  of  OBP and zinc-Induced testlcular atrophy  (Cater
et  al.,  1977).   Methylenedloxyphenol compounds  and  paraoxon Inhibited  DEHP
hydrolysis  by  rainbow trout   liver  In  vitro   (Melancon and  Lech,  1979).
Foster  et  al.  (1980)  did  not  find  antagonistic  effects  between  testlcular
zinc  levels  and  the   two  phthalates  DEP   and DMP.    In  rats  Initiated  with
04770
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dlethyln trosamlne,  the  administration  of   a   chollne   supplemented  diet
containing DEHP did not result  1n  Increased  hepatic  preneoplastlc  foci.   The
administration  of  a  chollne deficient  diet  containing  DEHP  Inhibited  the
appearance of preneoplastlc  fod.  Chollne deficiency  has  been  shown  to  be a
promoter  of   hepatic   preneoplastlc   foci   but  the  mechanism   of   DEHP's
ant1-pronot1ng  effect  Is  unclear, according  to  the  Investigators  (Deangelo
and Garr?tt,  1983).   There were no synerglstlc  or  antagonistic Interactions
found foi  B8P 1n the available literature.

    The  interaction between  ethanol  and DEHP has been  studied  by  Agarwal  et
al. (198;'b).  After a  single oral  or  l.p.  dose of DEHP to mice, the ethanol-
Induced  sleeping  time was   Increased  while  hepatic  alcohol  dehydrogenase
activity  was  Inhibited.   Repeated administration,  however,  produced  effects
that  differed  with  the  route  of administration.   When  DEHP  was given  In
repeated  oral  doses,  the ethanol-lnduced sleeping  time  was decreased,  but
Increase; In  the activities  of  both  alcohol  and  aldehyde  dehydrogenases  were
observed.   Repeated  l.p.  doses,  however,  resulted  1n  an  Increase   1n  the
sleeping  time  with  decreased alcohol  dehydrogenase  activity.   The  authors
concludec  that  DEHP  effected  changes  In   the pharmacologlc  response  to
ethanol  Dy  altering  the  activities  of  alcohol dehydrogenase and aldehyde
dehydrogtnase.   .In. vitro assays with mouse  liver preparations  revealed  that
MEHP  Inhibited alcohol dehydrogenase  activity.   Furthermore,  both MEHP and
DEHP  1nh  bUed  to  a statistically significant (p<0.05) degree  the  activities
of  both Mgh  and low Km aldehyde dehydrogenase activities.

Enzyme  Irdudnq Properties
    Most  studies of the  mechanisms  of  toxIcHy  have  researched  the effects
of  PAEs  on  enzyme systems  and metabolites.   Pollack and  Shen  (1984)  used
04770                              VII-2                             07/02/91
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antlpyrlne  metabolism  as a  model  for metabolic  clearance  of  drugs.   Ant1-
pyrlne's metabolism was  Increased  In  normal  and  renal  failure rats  (Sprague-
Dawley  rats In which  renal  failure  was  Induced by a  two-step nephrectomy)
after  treatment with  DEHP.   The plasma clearance was  Increased and  elimina-
tion   half-life   of   antlpyrlne  decreased   upon  DEHP  administration.    An
Increase In the  liver  weight and cytochrome P-450  content  was  also  noted as
evidence of  Induction  of hepatic mlcrosomal enzymes by  DEHP.   Renal  failure
rats appeared  to  undergo  a more marked Increase  1n  antlpyrlne clearance than
did control animals after DEHP treatment.

    Changes  In  hepatic enzyme activities  are associated  with liver  enlarge-
ment and occur In  animals exposed  to  PAEs  (Seth, 1982).  One change  that has
been observed  consistently  following  oral  or 1.p.  administration of  DEHP Is
a  decrease  In  hepatic  sucdnate  dehydrogenase   (SDH)  activity  occurring
specifically 1n the perlportal zones  {Seth,  1982).
    One  target  site for PAE  effects  on the liver  1s  mitochondria.   Results
of  \n_  vitro studies have  Indicated  that several PAEs  produce  Inhibition of
mitochondria! respiration.  It has been  suggested  that  PAEs  are electron and
energy transport  Inhibitors,  and  that  they  can  cause  uncoupling of oxldatlve
phosphorylatlon   (Seth,  1982).   Since  DEHP  Inhibited  the  activities  of
succlnlc  dehydrogenase (SDH)  and adenoslne trlphosphatase  (ATPase)  1n  rat
heart,   lung,  kidney  and  gonads as   well  as  the  liver,  suppression  of
energy-linked reactions may  be a generalized effect of  DEHP.   The enzymatic
alterations  may  not  be  related  to  the  physical  presence  of   DEHP  since
effects  were  present several  days after  final  treatment, by  which  time the
plastldzer would have been excreted from the body.
04770
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    DEHP  ias  also been shown  to affect carbohydrate  metabolism.   Decreased
levels  of  glycogen were  reported 1n  the  livers of  mice,  rats and  ferrets
receiving  DEHP   {Seth,  1982).    Marked depression  of  glucose and  glycogen
levels  was  found 1n  the  livers  of  rats  fed diets  containing 2 or  4% DEHP
(Sakural  ;t  al.,  1978).   Glucogenesls and glycogenolysls are  also  Inhibited
by  DEHP.   However,  no  quantitative  conclusion  on  the Inhibition  of  the
reaction .) upon hepatic  enzymes,  Upld  peroxldatlon  and hepatic sulfhydryl
content  In  rats.   The authors  concluded  that  the PAEs  Interfered  with  b1o-
transformetlon  mechanisms  of  hepatic  mlcrosomal drug-metabolizing  enzymes.
After a  single  oral  or  1.p. treatment of  DEHP was administered  to  rats,  the
activity of am1nopyr1ne-N-demethylase  and  aniline hydroxylase  was Inhibited.
When DEHP was given  In  repeated  doses,  the results  showed Increases In these
enzymes  with  oral  administration   but  decreases with  l.p.  Injection.   The
activity  of  benzo[a]pyrene  hydroxylase   and  concentrations  of  cytochrome
P-450 wens  also Increased 1n  rats  that  were treated orally with DEHP.   The
differences 1n  the effects  from  oral and  l.p.  administrations  may be attrib-
utable  to  variations  1n  the  physical state  and metabolism  of DEHP  after
Introduction of the compound Into the Intestine and  the peritoneal cavity.

    Studies by  Seth  et  al. (1981)  also Indicated that  the activities  of  the
liver  am1iopyrtne-N-demethylase  and  aniline hydroxylase  vere  Inhibited  by
l.p. administration  of  DMP, DEHP and  DBP to rats.  Hepatic  tyroslne  amlno-
transferase  activity was  unchanged after  a single  administration but  was
Increased when  the  PAEs  were given dally  for 7  days.   The authors  concluded
04770                              VII-4                             08/05/88
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that  these  results  are  supportive  of  previous  observations  that PAEs prolong
barbiturate  sleeping  time by Interference with  the  metabolic  disposition  of
these pharmacologlc agents.
    Walseth  et  al.  (1982} demonstrated contrasting results  of  PAE  treatment
on  rat  liver  and  lung.   DBP  administered  l.p.  resulted  1n  significant
Increases  In hepatic  cytochrome  P-450  but  reduced  lung  concentration  by
30X.   DMP  and DEHP were  less  effective  In this regard.   DBP  treatment  also
altered  the  enzymatic  pathways  of  benzo[a]pyrene  (B[a]P)  metabolism In "liver
mlcrosomes while  all   PAEs  tested  decreased  pulmonary metabolism  of  B[a]P.
These  authors did not detect a relationship between  carbon chain  length  of
PAEs and effects of mlcrosomal enzyme activities.
    While  DEHP  Is  associated  with  Increases  1n  the  activity  of  hepatic
mlcrosomal enzymes, Khawaja  and  Oallner  (1982)  determined  that liver protein
synthesis In  vivo  was  decreased after administration  of the  compound In the
diet  of rats.   Although  liver  weight  and  protein  content   Increased,  the
capacity for  treated  livers  to synthesize protein was  reduced.   The accumu-
lation  of  protein was  explained  as a  result  of   reduced  degradation  or
decreased export of liver proteins.
    There were  no available  data  concerning the enzyme  Inducing  properties
of EBP and OEP.
Cellular Effects
    Ekwall et  al.  (1982)  assayed  29 plastlclzers  Including DMP,  OEP,  DBP,
DEHP,  BBP and  three  other PAEs,  for   cytotoxldty  of  HeLa cells.   Cyto-
toxlclty  was  measured  by  pH changes  of the medium  using phenol red  as  the
04770
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07/28/88
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Indicator and by microscopic  Inspection of  the  cultures  (the HIT-24 system).
A  comparison  of  the  results  of  this  Vn  y 11ro cytotoxldty  test  to  other
cytotoxlc ty tests demonstrated  that  as the chain lengths  of  PAEs Increase,
llpophlllc Hy Increases.   A comparison of  these ^n vitro  cytotoxldty  test
results  with  ^n  vivo  test results  In  mice  suggest  that  a basal  cytotoxlc
action to mouse tissues  1s  responsible  for  the  lethal  action of plastldzers
to mice.

    The  c;'totox1c mechanisms  of  PAEs  may  be  better elucidated  by  studies  of
subcellulir distribution and activity as opposed to assays  of  tissue distri-
bution  (Bsll,  1982).    Bell  (1982) discussed  a series  of   experiments  con-
ducted In  rats, rabbits and pigs that  were directed at  the Investigation  of
PAE effee :s  on  llpld  metabolism.   In  studies of rats and  rabbits  that  were
fed  OEHP,   the  dlester  Impeded  cholesterol  synthesis  by  Inhibition  of
3-hydroxy-3-methylglutaryl  CoA reductase, which  catalyzes the  second step  of
cholestercl  synthesis.   The  effect  was  neither  sex  nor  species-specific.
Similar  Inhibition  of   cholesterol  synthesis  was  found   to  occur  In  the
adrenal  glands  and  testes.   Such Impairment  of   cholesterol  synthesis  In
these tls-. ues was thought  possibly to account  for fetal abnormalities  found
In the of:spr1ng  of phthalate-treated dams, and testlcular  atrophy  In  other
animals.   Plasma  and   liver  cholesterol levels  were  decreased  1n  rats  fed
either OBf  or DEHP.  Inhibition  of  cholesterol  synthesis by these  esters may
have been  the  underlying cause  for  this  effect.  Experiments with  jjn  vitro
tissue   slices  of  rats fed  DEHP  demonstrated  that  cte  novo  fatty  add
synthesis  and   esterlf1cat1on  are  Inhibited  In  certain  tissues  after PAE
administration.    Phosphollpld synthesis  may  also be  selectively  affected
(Bell, 19£2).
04770
VII-6
08/05/88
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    Upon  further  Investigation Bell and Buthala  (1983)  discovered  that DEHP
 Inhibits  mlcrosomal  acylCoArcholesterol  acryltransferase (ACAT)  In  rats that
 received  this  compound  1n  the diet.  The  biosynthesis  of  cholesterol  from
 14C-mevalonate  was  also  Inhibited  1n  treated animals Indicating  that  other
 mlcrosomal  enzymes   are  Influenced  by  DEHP  administration.    The  post-
 mevalonate  segment  of  the  Dlosynthetlc  pathway requires  the  Involvement  of
 numerous  mlcrosomal  enzymes,  while  cholesterol  esterlfIcatlon Is  largely
 associated with ACAT.
    Bell  (1982)  also  4escr1bed experiments performed orv  the  effects  of DEHP
on mitochondria!  function.   Administration of DEHP  to  rats  (50,  250 and 500
mg/kg/day  In  the  diet  assuming  rats  consume  5%  of  their  body  weight),
rabbits  (490  mg/kg/day In diet  assuming rabbits consume  4.9%  of  their body
weight)  and  pigs  (1.6  mg/kg/day)  resulted  In   Increased  production  of.
palmitic  acid  by  liver  mitochondria  accompanied  by  an  enhancement  of
14C-palm1toyl  CoA  oxidation.    Studies  on  heart  mitochondria  demonstrated
that  DEHP directly  added to  an  Isolated suspension  will   Inhibit  aden'.ne
nucleotlde  translocase.   Thus,   exchange  of  extramltochondrial  ADP  for
1ntram1tochondr1al  ATP  Is  Impeded.   Inhibition  of  translocase  was  not
observed  In   the  mitochondria  Isolated  from rats  fed  DEHP compared  with
controls.   It was  felt  that  a  level  of  DEHP  Insufficient  to  affect  the
enzyme had  accumulated 1n the  heart  during  the 10-day  feeding period.  The
author concluded  that  Inhibition of  heart  adenlne  nucleotlde  translocase may
be related to reports  of myocardlal cell  death  and  decreases In spontaneous
heart  rate  observed 1n  rat  hearts after  DEHP perfuslon  (Rubin  and  Jaeger,
1973; DeHaan, 1971; Petersen et  al.,  1972-1975;  Aronson et'al., 1978).  Bell
(1S82)   noted  that  the  biochemical   transformations   observed   1n  these
experiments   Indicated   that    the   effects  of    PAEs   may  result   from
04770
VII-7
09/07/88
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alterations  of  membrane  fluidity.   The  UpophllU properties  of the  PAEs
may,  therefore,  change  the   membrane  environment  sufficiently  to  modify
enzyme responses.

    Melnlck and  Schiller  (1982)  studied  the effects of  DMP,  OBP  and  DEHP on
liver mitochondria  Isolated from rats.   Active transport of  potassium  Ions
(K*),  resjlratlon  rates  and  succlnate  cytochrome c   reductase  activities
were monitored.   D8P  was  the  most effective energy  uncoupler  as  measured by
Interference  with  Kf uptake  Induced  by  three  energy  sources.  It also  led
to  a  nearly  total  loss of  respiratory control.   DMP  was less  effective In
this  regard;  MEHP,  but  not the parent DEHP,  was  an effective  uncoupler of
energy-11n
-------
                                  TABLE  VII-1

                Cellular Changes In Rat Hepatocytes Induced by
                             DEHP Administration3
             Organelle
                       Changeb
Peroxlsomes
  Protein and phosphollpld
  Beta-oxidation enzymes of fatty adds
  Carn1t1ne-acetyl transferase
  Catalase
  Urate oxldase

Mitochondria
  Protein and phosphollpld
  B.eta-oxidation of fatty adds
  Carn1t1ne-acetyl transferase
  Carnltlne-octanoyl transferase
  Carnltlne-palmltoyl transferase
  Dehydrogenase and respiratory
  Respiratory control and oxldatlve
   phosphorylatlon

Mlcrosomes
  Protein and phosphollpld
  NADPH-cytochrome c reductase
  Cytochrome P-450
  Other electron transport enzymes,
   hydroxylases, phosphatases

Homogenate
  Sterol and squalene synthesis
  CoA and carnltlne
  Acetyl-CoA and acetyl-carnltlne
  Long chaln-acyl CoA and -acyl
   carnltlne
            Increased several-fold
            Increased 2- to 6-fold
            Doubled
            Decreased 30-40%
            Decreased 30-40%
            Increased 2- to 3-fold
            Doubled
            Increased 10- to 30-fold
            Tripled
            Increased 3- to 4-fold
            No or moderate change
            No change
            Slight Increase (10%)
            Increased 40-60%
            Increased 40-60%
            No change or moderate Increase
            Decreased 75%
            Increased 5- to 6-fold
            Increased 4-fold
            Increased 50%
aSource: Gannlng et al., 1984

^Specific activities or amounts on protein basis compared with the control
04770
VII-9
09/07/88
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                                  TABLE  VII-2

      Synthesis and Breakdown of Protein and L1p1d 1n DEHP-Treated Rats3

Protein
Peroxlscmes
Catalase
Beta-cxldatlon enzymes
Mltochordrla
Membrane
Beta-cx1datlon enzymes
Mlcrosonal membranes
Total cytoplasmlc
proteins
Amount or
Activity
Decrease
Increase
Increase
Increase
Unchanged
Unchanged
Synthesis
Decrease
Increase
Increase
Increase
Increase
Increase
Breakdown
Control -» Treated
Half-time 1n Days
1.9 -* 5.0
2-3 -» 5.5-6.5
6 -» 25
Decrease
3.5 -» 5.5
2.5 -» 5
Llpld
  Hlcrosonral phosphollplds     Unchanged
  Blood cholesterol
    Total                      Unchanged
    HDLb                       Decrease
    LOLC                       Increase
    VLDLd                      Unchanged
          Increase
aSource: Canning et al., 1984

bHDL, high-density Upoproteln

CLDL, low-density llpoproteln

dVLDLf very low-density Upoproteln
04770
VII-10
07/28/88
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oxidation of CoA-linked  fatty acids,  In mlcrosomal  NADPH-cytochrome  c  reduc-
tase and  cytochrome  P-450 levels,  In  the  number  of mitochondria and  In  the
activity  of  carnitlne-acetyl  transferase.   Induction of the  transferase  was
attributed to an  Increase  1n peroxlsomal B-oxidat1on.   Ganning  et al.  (1983)
demonstrated  that  although  peroxlsomal  and  mitochondria!  membranes  were
Increased, the endoplasmlc retlculum was  not changed In  amount or appearance.
    Increases  In  the activity  of  enzymes 1n  rat  hepatic cytosol  have  been
found upon  application  of  various  peroxlsome prollferators  Including  DEHP,
DAP  and  2,4,5-triphenoxyacetic  add  among  others  (Katoh   et  al.,  1984).
Administration of  DEHP  resulted in  the  induction  of catalase and  two  long-
chain acyl-CoA hydrolases.   An  Increase In peroxlsomal 0-oxidatlon  was  also
signaled by  a  marked increase 1n palmHoyl-CoA oxidation after  ingestlon  of
DEHP in the diet.

    Primary  rat   hepatocyte  cultures  were   used   to  ascertain  effects  of
various  alky!  phthalate  esters on  peroxlsomal  enzyme  activities   (Gray  et
al.,  1983).   The   authors  concluded that  straight-chain  phthalates  produce
few  effects  upon  rat hepatic  peroxisomes.   The  2-ethylhexyl  ester,  e.g.,
MEHP,  increased   carnitine  acetyltransferase  activity  and  palmltoyl   CoA
oxidation, and produced  Increased numbers of  peroxisomes.
    The effects  of  different PAEs upon  liver  cells have been  compared  with
those  of  clofibrate,  another peroxlsome  proliferator  (Lake et  al.,  1984b).
Lake et al.  (1984a)  had previously determined that DEHP  Is  a  potent  inducer
of  rat hepatic  peroxisomal  enzyme  activities.    In  the more   recent  study
(Lake  et  al.,  1984b),  rats   were   orally  administered  DEHP,  di-n-octyl
phthalate  (OOP),  mono-n-octyl   phthalate  (MOP) or  clofibrate  for 14  days.
04770
VII-11
09/07/88
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This resulted 1n liver enlargement.   Liver  sections  from DEHP and cloMbrate
treated animals  showed an  Increased  number of  peroxlsomes.   Both  DEHP  and
clofibrate   stimulated   the  activities   of   peroxlsomal  marker   enzymes,
Increased  mlcrosomal  cytochrome  P-45Q   content  and  stimulated  mlcrosomal
laurlc  ac  d  hydroxylatlon  activity.   The  compounds,  OOP and  MOP,  did  not
produce sich effects.  The  branched chain ester OEHP was  thus  determined to
exert  effects  that  differed markedly  from  the straight chain  analogue,  OOP
and  Us iretabollte  MOP.   In addition,  OEHP  was  shown  to  Induce  forms  of
cytochrome P-450  similar  to those  Induced  by  clofibrate.  Oklta  and  Chance
(1984)  al;o  demonstrated  that  DEHP,   like  clofibrate,   Increased  mlcrosomal
laurate hidroxylatlon  activities.   Potent  Induction of  the  cytochrome P-450
mediated  :atty  acid u-hydroxylatlon  reaction  occurred  In  rats that  were
fed a  diet containing OEHP.

    There may  be some species  variation  In  the  biochemical  actions  of PAEs.
Lake   et  <,1.  (1984a)  compared  DEHP,  MEHP and cloflbrate-lnduced  hepatic
peroxlsome   proliferation   1n   two  species,   rats   and  hamsters.    It  uas
discovered that  DEHP was  much  less  effective as a peroxlsome prollferator in
hamsters   than   In   rats.   Similar   results  occurred  when  clofibrate  was
utilized.  Although  all  three compounds  caused some  Increase 1n liver  weight
and  hepatic  peroxlsome numbers, the  response  was more  marked  in rats,  for
each  of the three  treatments,  dose  dependent Increases  1n  the peroxlsomal
marker,  cyanide-Insensitive palmHoyl-CoA, and  In  carnltlne acetyltransfer-
ase  were  loted  In   the  rats.   Only  small  changes  In  these  parameters were
found   1n  the  hamsters.   The species variation  In  the effects  of  DEHP  may
have  been  attributable to differences in  peroxlsome  proliferation  or  in the
metabolism of  OEHP.

04770                               VII-12                            09/07/88
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    The  mechanism  of carcinogeniclty for  DEHP  Is not well  understood;  how-
ever,  It  has  been  suggested  that  DEHP may  fall  Into the perox1some~prol1fer-
ator  class  of hepatocarclnogens  (Warren  et  al.,  1982).   Peroxlsomal  prolife-
rating effects  and hepatomegaly  do not  seem  to  be  related  to differences  1n
the sensitivity of  suckling  rats  to  toxic  effects caused by exposure to DEHP
(Dostal  et  al.,   1987a).    Changes  In   relative  liver  weight  and  hepatic
peroxlsomal  enzyme activities were  similar  In  age groups  showing  markedly
different changes  1n body weight  and  survival  rates.   Similar  Increases  1n
activities  of  both  palmitoyl CoA oxldase  and   carnltlne  acetyltransferase
were noted  between  suckling  and adult rats  Indicating that  suckling rats are
equally  If  not more  sensitive  to the  peroxlsomal  proliferating  effects  of
DEHP  {Postal  et al.,  1987a)  (see Table V-4).  The  Induction of perloxlsomes
and peroxlsomal  enzyme  activity  as  well  as hypolipldemlc  effects was  not
delected  In marmoset  monkeys exposed  eHher orally (2000 mg/kg/day)  or  l.p.
(1000 mg/kg/day) to  DEHP  {Rhodes  et al.,  1986).   Also,  there was no Increase
In  cyanide-Insensitive  acyl oxldase,  the  peroxlsome   marker  enzyme.   The
marmoset  appears   to  be  less  sensitive  to the peroxlsomal  proliferating
effects  of  DEHP.   Rhodes et  al.  (1986)  concludes  that  If  marmosets  reflect
more  accurately the response in  man, then  low  levels of DEHP may not  be  of
to;<1colog1c significance with regard  to  hepatocellular carcinoma.
    Possible mechanisms  for  the hepatocarclnogenlc effects  of  phthlates  and
other peroxlsome prollferators  have  Included  the  generation  of  free radicals
from   Increased   hydrogen   perloxlde   (HO)    production   and   decreased
catalase  activity,  and  that   peroxlsome   Inducing  chemicals  and/or  their
metabolites may act  as  promoters  (Gannlng et al., 1984).   The  production of
the  enzyme  catalase by  peroxlsomes  catalyzes  the   breakdown  of  hydrogen
04770
VII-13
07/02/91
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peroxide to  water.   Hydrogen  peroxide  Itself or  the  hydroxyl 1on,  that  Is
formed  from  hydrogen  peroxide,  causes   damage   to   DNA   and  chromosomes
(Turnbull  and  RodrUks, 1985).   PAEs,  such  as  DEHP,   exhibit  hypollpldemlc
activities   common   to  several  peroxlsome  pro!Iferators  that   Include  liver
enlargement  that  Is   not   accompanied  by  frank  hlstologlc  Hver  damage,
proliferation of smooth endoplasmlc  retlculum and  an Increase  1n  the number
of  hepat c   peroxlsomes  (Cohen  and  Grasso,  1981).  Warren  et  al.  (1982)
hypothesised that  If  DEHP  acts similarly to  other  peroxlsome  prollferators,
the compound may Initiate  neoplastlc transformations of  hepatic  parenchymal
cells by Increasing Intracellular  reactive  oxygen  species,  which  could cause
DNA  damage.  Peroxlsome  prollferators  modify  peroxlsomal  enzyme  profiles
such   thct   fatty  add   B-oxIdatlon,   H2°?'   peroxldlzed   llpofusln   and
peroxlsomil  uMcase levels  are Increased and  Increased catalase  activity  1s
smaller with respect  to the Increased peroxlsome volume.   That  Is,  catalase
activity Is  Increased, but  to a much lesser  extent than  the  activities  of
H_0. gene~at1ng oxldases.

    Turnbill  and   Rodrlcks   (1985)   proposed  a   possible   mechanism   of
cardnogeildty  for  DEHP   based  on  a   peroxlsome   proliferation  hypothesis
(Figure Vll-l).  As discussed  1n  Chapter III,  the  Initial step 1n metabolism
of  orally  administered OEHP  Is  hydrolysis  to yelld MEHP.   MEHP  then under-
goes  w-  and  w-1-oxidation.    One  of  the  ^-oxidation  products  may  then
undergo  3-ox1dat1on,  which   1s   Important   since   In   this  step  of  DEHP
metabolism  hydrogen peroxide  Is  generated.   Excess Intracellular  levels  of
hydrogen peroxide  may  be  detrimental to  the cell.   In addition,  hydrogen
peroxide :an react  with DNA,   causing alteration and liberation of DNA bases
and sugar-phosphate backbone breakage (Turnbull and Rodrlcks, 1985).

04770                              VII-14                            07/02/91
-------
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04770
                                                 VII-15
                                                       07/28/88
-------
    In  a  more  recent  report  RodMcks   and  Turnbull  (1987)  compared  and
summarized  the  differences between  peroxlsomes found  1n various  mammalian
species.  The  most  extensive studies  on proliferation  of  peroxlsomes  and
Inductlor  of peroxlsomal  enzymes  have been 1n male rats.  Species  differ  In
their morphologic  characteristics  of  peroxlsomes.   Humans, as well  as  other
species  lack  the  enzyme  uric  acid  oxldase  (urlcase)  since  the  central
crystallcld  core  Is  absent from the  peroxlsome.  When  comparing  peroxlsomal
data,  th»re seem  to  be  only slight  differences  between species;  however,
there  are  even  some   differences  within  a   species   relating  to  age  and
gender.   Of  the species and  sexes  tested, male rats are  the  most  sensitive
to  chem1:ally   Induced  peroxlsomal  proliferation.   Quantitative  measurement
of  the  sjecies  differences  Is  not  available.   However,  the authors  speculate
that  H  may  be due  to  differences  1n  absorption,  metabolism or  Inherent
differences  1n hepatic susceptibility (RodMcks and Turnbull,  1987).

    The possible DNA-blndlng  activity  of  DEHP has  been  Investigated by  Albro
et  al.  |1983a).   Ethylhexyl-labeled  DEHP,  but not  ring-labeled  DEHP,  was
found  to  be associated with  the  DNA  from  the  livers  of  rats.   The authors
determined  that the  radioactivity  was  not a result  of  absorption,  Intercala-
tion,  attachment   to  RNA  or  hlstones, an  Impurity  1n  the  labeled  DNA,  or
artlfactial  binding   from  the  sample preparation.  The  source  of  the  14C
may  have  been  carbonyl  phosphate, which  Is a precursor for  urea  and pyrlmi-
dine  basi;s.  von  Oanlken et  al.  (1984)  concluded  that  DEHP did  not  bind
covalently  to  hepatic  DNA  1n rats  and  mice  exposed  to  the labeled  PAEs
through  dietary administration.   Radioactivity  associated with  the DNA was
attributed   to  the  biosynthetlc  Incorporation  of  radlolabeled  breakdown
products, such  as  2-ethylhexanol.


04770                              VII-16                             07/02/91
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Mechanisms of Reproductive ToxIcUy
    Gonadal  toxlclty  1n  rats  has  been  linked  to the  adverse  effects  of
phthalates upon   testlcular  zinc  concentrations.   Upon administration of DBP
or  DEHP  urinary  excretlor of zinc was  enhanced  and the testlcular zinc con-
tent  decreased   (Cater  et al.,  1977;  Foster  et  a!.,  1980;  Thomas  et  al.
1982).   Cater et  al.  (1977)  concluded that  after oral  administration, DBP 1s
metabolized  by   nonspecific   esterases  In  the  GI  tract   to  the  monobutyl
phthalate  (MBP)  prior  to  absorption  Into the  bloodstream.   The  monoester or
another  metabolite  of  DBP may act as a chelatlng  agent by removing the zinc
from  the  testes.   Testlcular  zinc   deficiency  Is, therefore,  the possible
causative  factor  leading  to  testlcular  atrophy.   Z1nc  depletions  have been
noted  In both the testes  and  prostate glands  of  rodents following oral, s.c.
and  l.p.  PAE  exposures.   It  has  been   hypothesized  that   the  testlcular
effects  of orally administered dlesters  are  mediated   by  the  monoesters and
alcohols  produced  during dlester   hydrolysis  1n  the  GI   tract   {Gray  and
Becmand, 1984).   Thomas et  al.  (1982) provided  s.c. and l.p.  Injection data
that  demonstrated  the action  of  DEHP  upon   the  depletion  of  endogenous
goriadal  zinc was  not  a  function  of  the  Interference  of   the  Intestinal
absorption of the divalent zinc 1on.
    Further  Investigations  of the  mechanism of  testlcular  Injury  Indicated
that the testlcular  Injury  Induced  by  DBP  does  not appear to result from the
accumulation of  metabolites or the  formation  of covalent adducts  In  testl-
cular  tissue  (Gangolll,  1982).   Oral   administration  of  14C-OBP  (lactation
of  »*C  not stated)  did not  show  evidence of  accumulation  of  radioactivity
In the gonads.   Also,  testlcular atrophy did not appear  to be mediated by an
Interference  In  androgen  synthesis  or  the  availability  of  gonadotroplns.
04770
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The  DBP-Induced  testlcular  Injury  was   not  reversed  by  treatment   with
testosterone or pregnant mare serum (Gangolll, 1982).

Summary
    Research  Into  the  mechanisms  of  PAE  toxldty  1n animal  tissues  has
Indlcatec  that the PAEs may  Interfere with  the  normal  enzymatic  or  metabolic
processes.  Investigators have  found  that  PAEs  exert  their toxic effects  by
modifying   the  physical  state   of  membrane-1tplds  and,  therefore, change
membrane fluidity.  The mechanisms  by  which these alterations occur  has  not
been clearly  delineated.   In the  liver,  phthalates  alter  the structure  and
metabolism  as  characterized by  Increases  1n   the  number  of  peroxlsomes,
mHrochondrla  and  enzymes  of   fatty  add  oxidation.   Studies primarily  on
DEHP  Indicate  that   llpld   and  protein  metabolism  are  Inhibited.   These
effects en  carbohydrate metabolism are also  associated with  depressions  1n
the  energy  coupling   systems   of  the  liver,  Including  the mitochondria.
Inhibition of  cholesterol synthesis In various  organs  occurs  when  phthalates
Inhibit 
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                 VIII.  QUANTIFICATION OF TOXICOLOGIC EFFECTS

Introduction
    The  quantification of  toxicologlc  effects  of  a  chemical  consists  of
separate  assessments  of  noncarclnogenlc  and carcinogenic  health  effects.
Chemicals  that  do not  produce  carcinogenic  effects  are believed to  have  a
threshold  dose  below  wh'ch  no adverse,  noncarclnogenlc  health  effects  occur,
while carcinogens are assumed to act without a threshold.
    In  the  quantification  of   noncarclnogenlc   effects,  a  Reference  Dose
(RfD),  [formerly  termed  the  Acceptable Dally  Intake (ADI)]  Is  calculated.
The RfD  Is an  estimate (with  uncertainty  spanning perhaps an  order  magni-
tude)   of   a  dally  exposure  to the  human  population   (Including  sensitive
subgroups) that  Is  likely  to be  without  an appreciable  risk  of  deleterious
health effects  during a  lifetime.   The RfD  Is  derived  from  a  no-observed-
adverse-effect   level   (NOAEL),   or    lowest-observed-adverse-effect   level
(LQAEL),  Identified  from a  subchronic  or  chronic  study, and divided  by  an
uncertainty factor(s)  times a  modifying  factor.   The  RfD Is calculated  as
follows:
     RfD =
                        (NOAEL or LOAEL)
           [Uncertainty Factor{s) x Modifying Factor]
                        mg/kg bw/day
    Selection of the uncertainty factor  to  be  employed  In the calculation of
the RfD  Is based  upon  professional judgment,  while considering  the  entire
data base  of  toxicologlc  effects  for the chemical.   In  order  to ensure that
uncertainty  factors  are  selected  and applied  In a  consistent manner,  the
04780
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U.S.  EPA  (1991)  employs  a modification  to  the  guidelines proposed  by  the

National Academy of Sciences (NAS, 1977, 1980) as follows:


Standard Uncertainty Factors (UFs)

        Lse a 10-fold factor when  extrapolating  from valid experimental
        results from studies using prolonged exposure to average healthy
        humans.   This  factor  1s  Intended  to account for  the  variation
        1n sensitivity among the members of the human population.  [10H]

        Lse an  additional  10-fold  factor  when extrapolating  from valid
        results  of  long-term  studies  on  experimental  animals  when
        results of  studies  of  human  exposure  are not available  or  are
        Inadequate.  This  factor  Is  Intended to  account  for  the uncer-
        tainty  In  extrapolating  animal  data to the  case of  humans.
        [IDA]

        Use an  additional  10-fold factor  when  extrapolating  from  less
        than  chronic  results  on  experimental  animals when there  Is  no
        useful  long-term  human  data.   This  factor  1s  Intended  to
        account  for  the  uncertainty  In  extrapolating  from   less  than
        chronic NOAELs to chronic NOAELs.   [10S]

        Use an  additional   10-fold  factor  when deriving  an RfD  from  a
        LOAEL  Instead of  a NOAEL.   This  factor   Is  Intended  to account
        for  the  uncertainty  1n  extrapolating  from  LOAELs to  NOAELs.
        [10L]

Modifying Factor (MF)

        Use  professional  judgment  to  determine  another  uncertainly
        factor  (MF) that Is greater  than  zero and less  than or equal  to
        10.   The   magnitude of  the  MF  depends  upon the  professional
        assessment  of  scientific  uncertainties  of  the  study  and  data
        base  not  explicitly treated above,  e.g.,  the  completeness  of
        the overall  data base  and  the number  of species  tested.   The
        default value for the MF 1s  1.
    The  uncertainty  factor  used  for  a  specific  risk  assessment  Is  based

principally  upon   scientific   judgment   rather   than  scientific  fact  and

accounts   for   possible   Intra- and  interspedes   differences.   Additional

considerations  not  Incorporated  In  the  NAS/OOW guidelines  for  selection of

an  unceralnty factor  Include the  use  of  a  less  than  lifetime  study  for

deriving  an RfD,  the  significance of  the  adverse  health  effects  and  the

counterbalancing of beneficial effects.
04780
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07/02/91
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    from  the RfD,  a Drinking  Water  Equivalent  Level  (DWEL) can  be  calcu-

 lated.   The  DWEL   represents  a  medium  specific  (I.e.,  drinking  water)

 lifetime  exposure at which  adverse,  noncarclnogenic health  effects are not

 anticipated  to  occur.   The  DWEL  assumes  100% exposure from  drinking  water.

 The DWEL  provides the noncarclnogenic health  effects  basis  for  establishing

 a  drinking  water standard.   For  Ingestlon  data,  the  DWEL  1s derived  as

 follows:
               DWEL
               (RfD)  x  (Body  weight  1n  kg)
              Drinking  Water  Volume  1n  a/day
     mg/i
where:
        Body weight = assumed to be 70 kg for an adult
        Drinking water volume = assumed to be 2 l/day for an adult
    In addition  to the RfD  and  the DWEL, Health Advisories  (HAs)  for expo-

sures  of  shorter  duration   (1-day,  10-day  and longer-term)  are  determined.

The  HA values  are  used  as  informal  guidance to  municipalities  and other

organizations when  emergency spills or  contamination  situations  occur.   The

HAs are calculated using an  equation  similar to the  RfD  and DWEL; however,

the NOAELs  or  LOAELs are identified from acute or  subchronlc  studies.   The

HAs are derived as follows:
                   HA =
                (NOAEL or LOAEL) x (bw)
                  (UF) x (	 i/day)
mg/s.
    Using the above equation,  the  following  drinking  water  HAs  are developed

for noncarclnogenic effects:
    1.
    2.
    3.
    4.
04780
1-day HA for a 10 kg child ingesting 1  l water per  day.
10-day HA for a 10 kg child ingesting 1  i water per day.
Longer-term HA for a 10 kg child Ingesting 1  1 water per  day,
Longer-term HA for a 70 kg adult ingesting 2  l water per  day.
                             VIII-3
              03/30/88
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    The  1-day  HA  calculated   for  a  10  kg  child  assumes  a  single  acute
exposure  to  the chemical and  1s  generally derived  from  a study of  <7  days
duration.  The  10-day  HA assumes  a limited exposure  period of 1-2  weeks  and
Is generally  derived  from a study of <30  days  duration.   The longer-term HA
Is  der1v;d for  both  the 10  kg  child  and a  70  kg  adult  and assumes  an
exposure  period  of  -7  years   (or  10%  of  an  Individual's  lifetime).   The
longer-term  HA  1s  generally  derived  from a  study  of   subchronlc  duration
(exposure for 10% of animal's lifetime).

    The L.S.  EPA  categorizes the  carcinogenic potential of a  chemical,  based
on the overall welght-of-evldence, according to  the following scheme:

        Group  A:  Human   Carcinogen.   Sufficient  evidence exists  from
        epidemiology  studies   to  support a  causal association  between
        exposure to the chemical and human cancer.
        Group  8:  Probable  Human  Carcinogen.    Sufficient evidence  of
        carclnogenlclty  In  animals with  limited  (Group  Bl)   or  inade-
        quate (Group B2)  evidence 1n humans.
        Group  C:   Possible  Human   Carcinogen.    Limited  evidence  of
        carclnogenldty 1n animals In the absence of human data.
        G'oup  0:  Not  Classified  as to  Human  Carclnoqenlclty.   Inade-
        qjate human and  animal  evidence  of  cardnogenldty or for which
        nD data are available.
        G'oup  E:   Evidence   of  Noncarclnoqenlclty  for   Humans.    No
        evidence  of cardnogenldty  1n  at  least  two  adequate  animal
        t?sts  in different  species  or   In  both  adequate  epldemlologlc
        aid animal studies.

    If  tcxlcologlc  evidence leads  to  the classification   of  the contaminant
as a  known,  probable or  possible  human carcinogen, mathematical models  are
used  to  calculate  the  estimated  excess  cancer  risk  associated  with  the
Ingestlon  of  the  contaminant   In  drinking water.   The   data  used  In  these
04780                                VIII-4                          07/02/91
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 estimates  usually  come  from  "lifetime  exposure  studies  using  animals.   In
 order  to predict the risk  for  humans  from animal data, animal doses must  be
 converted  to  equivalent  human  doses.   Thl.s conversion  Includes correction
 for  noncontlnuous exposure,  less  than  lifetime  studies  and for differences
 In  size.  The  factor  that compensates  for  the  size  difference  Is  the cube
 root  of  the ratio of the  animal and human body weights.  It 1s  assumed that
 the  average adult  human  body  weight  Is  70 kg  and  that  the  average  water
 consumption of an adult human  1s 2 l of  water per day.
    For  contaminants  with  a  carcinogenic  potential,  chemical  levels  are
correlated  with  a carcinogenic  risk  estimate by  employing  a cancer potency
(unit  risk} value  together  with  the  assumption  for  lifetime  exposure  from
1nc;est1on  of  water.   The cancer unit  risk  Is  usually derived from a linear-
ized multistage model  with  a 95% upper confidence limit providing a low  dose
estimate;  that  Is,  the  true risk  to  humans, while  not  Identifiable,  Is not
Hkely  to  exceed the  upper  limit  estimate  and,  In  fact,  may  be  lower.
Excess cancer  risk  estimates may also be calculated using other models  such
as  the one-hit,  Welbull,  logH and  probH.   There  Is  little  basis  In the
current  understanding  of   the   biologic  mechanisms  Involved  In  cancer  to
suggest that any  one  of  these models  Is  able to predict risk more accurately
than any other.   Because each model  Is based upon differing assumptions, the
estimates derived for each model can differ  by several orders of magnitude.

    The scientific  data base  used to calculate and support  the setting of
cancer  risk rate  levels has  an  Inherent  uncertainty  that  Is  due to  the
systematic and random  errors  1n  scientific measurement.   In  most cases,  only
studies  using experimental  animals  have been  performed.   Thus,   there  1s
04780
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07/02/91
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uncertainty  when   the  data  are  extrapolated  to  humans.   When  developing
cancer risk  rate  levels,  several other  areas of uncertainty  exist,  such as
the  Incomplete  knowledge  concerning  the health  effects  of contaminants  1n
drinking  .later,   the  Impact   of  the  experimental  animal's  age,  sex  and
species,  1 he  nature of the  target  organ  system(s) examined and  the  actual
rate of e) posure  of the  Internal targets 1n  experimental  animals  or  humans.
Dose-response data  usually  are  available  only for  high  levels of  exposure
and  not  for  the  lower levels  of  exposure  closer  to where a standard  may be
set.   Whei  there  1s  exposure  to   more  than  one  contaminant,  additional
uncertainty results  from a lack  of  Information  about  possible  synerglstlc or
antagonistic effects.

Noncarcinogenic Effects
    PAEs  
-------
OEHP.  A  second adult  male  subject  given  an oral dose of 10 g of OEHP exper-
ienced  mild gastric  disturbances  and  moderate  catharsis  (Shaffer  et  a!.,
1945).  Accidental  Ingestlon of  10 g  of  DBP by a young  adult  male produced
nausea, vertigo  and signs of  keratltls  and toxic nephritis  (Lefaux, 1968).
A  single  prospective  cohort  study was Identified  1n  the  literature.   Thless
et al. (1978b) reported  that among  221  workers  exposed to DEHP, only half of
the  expecte4 deaths were observed  In  the exposed population.   Although  the
analysis was not conclusive,  no  Increased  risk  of adverse health effects  was
attributed to exposure to DBP  In  this  group of  workers.   £n vitro studies of
human tissue and cell  cultures revealed that PAEs  Inhibited  cellular growth
and  decreased  platelet  function  but  did  not  produce chromosomal  damage In
human leukocytes or fetal lung  cells.   The greatest  risk  for  toxic  effects
from  PAE  exposure  appears to  be  among Individuals receiving  blood  transfu-
sions or  hemodlalysis  due to  extraction  of PAEs from plastic  blood  bags or
plastic tubing used In  these  treatments.   However,  reports of  hepatitis In
hemodlalysis patients  and necrotlzlng  enterocolHIs  In   Infants  given  blood
transfuslor  could  not be attributed definitively to PAE  exposure.

    Species  differences  occur with respect  to  metabolism  of  PAEs.   Several
species of animals  have  been determined  to excrete glucuronlde conjugates of
MEHP  (the major metabolite of  DEHP) upon  exposure to  DEHP with the exception
of rats (Tanaka  et  al.,  1975; Williams and  Blanchfleld,  1975;  Albro  et  al.,
1982).  The  role that glucuronlde  conjugation  may  play  In the  sensitivity
between species  to toxic  endpolnts  Is  not known;  therefore, studies  with
rats will  still  be  considered with other  test  species  for  quantification of
toxicologic effects.
04780
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    Thert  1s  no  common toxic effect  that  PAEs  as a group  of  compounds have
been shoeen  Identified for  PAEs, the HAs,  OWELs  and  cancer  risk  levels are
calculated  for  Individual   PAEs  rather  than  for  the  group  of  compounds
generlcally.

     Studies  Considered for  Noncardnoqenlc   Quantifications  — DEHP.   DEHP
has  been  studied more extensively than  any  other  PAE, In part because  H Is
the  most  widely used plastldzer.   Enlarged  liver  and  testlcular  atrophy are
the  two  most  commonly observed  effects of  DEHP  1n  rats.   Mangham et  al.
(1981)  conducted  a  short-term  test  to  examine  the   testlcular and  hepatic
effects cf  DEHP.   DEHP was administered  orally  by gavage to Wlstar rats at a
dose level  of 2500 mg/kg/day for 7 and  21 days.   After 7 or  21 dally doses,
weight  of  the testes  was  decreased,  and hlstopathologlc  changes  were  found
In  50-80}  of  the  seminiferous  tubules of  each  male  rat.  Treatment for 7 or
21   days  produced  marked  liver  enlargement  In  rats  of both   sexes  and
decreased  activity  of  succlnate dehydrogenase  1n males.   Also,  body weight
gain was  significantly decreased In  males.   The testlcular effects observed
are  cons'stent with  results  of an  earlier   study by  Gray et  al.  {1977} 1n
which  testlcular atrophy  occurred  within the first 2  weeks of  treatment at a

04780                                 VIII-8                          07/02/91
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dietary  level  of  2%  DEHP  (-1440  mg/kg/day).   Lake  et  al.  {1975}  reported
Increased  liver  weights  In  Wlstar rats  administered 2000  mg/kg  DEHP  [236
mg/kg/day assuming 0.013  kg/day  Intake  (Lehman,  1959)] for periods of 4,  7,
14 and 21 days.  The Investigator did  not  examine  any  reproductive  organs.

    In a recent  study,  MHchell  et al.  (1985) observed  similar  results  when
groups of  male and female  Wlstar albino  rats  were administered  diets  con-
taining 50, 200 and 1000  mg/kg/day DEHP for 3,  7,  14  and 28  days.   There was
a  total  of 90  treated  rats and  60  control  rats.  H1stopatholog1c  examina-
tions were performed on  the major abdominal  organs at all time  points.   The
livers of male  rats were significantly  enlarged 3  days  after  treatment  with
10CO mg/kg/day OEHP.  After  14  days  significant liver enlargement  was  noted
at the 50  and 200 mg/kg/day doses  1n  male rats.    There were  no significant
differences In testes  weight when  control  animals  were compared  with  experi-
mental animals.  Further  details were not  given.   Liver  cells  from male  rats
showed marked  proliferation  of  peroxlsomes after  3 days treatment with  200
or 1000 mg/kg/day.  Treatment with 50  mg/kg/day showed  Increased  numbers  of
peroxlsomes  after  14  days.  Female  rats,  however,   showed  only  Increased
number of peroxlsomes  after 14 days treatment with 1000  mg/kg/day.   Prolife-
ration  of   the  smooth   endoplasmic  retlculum  In  both   males  and  females
occurred  at all  doses  In a dose-dependent manner.  Biochemical  changes  such
as effects  on DNA, catalase activity and laurate  hydroxylase activity  were
also noted  In all dose groups.
    SubchronU oral  studies  have been conducted  with  DEHP on  rats  (Shaffer
et  al.,  1945; Harris  et al.,  1956;  Nlkonorow  et al.,  1973;  Gray et  al.,
1977;  Mitchell  et al.,  1985;  Cater  et  al.,  1977), mice (NTP, 1984a),  and
04780
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dogs  (Hcrris  et   al.,  1956).   The  study  1n   which  adverse  effects  were
observed at  the lowest  level  of exposure  Is  that  of Mitchell  et  al.  (1985).
Mitchell  et  al.   (1985)   fed   male  and   female  WUtar  albino  rats  diets
contalnirg  50,  200 and  1000  mg/kg/day DEHP  for  9 months.  Necropsy  of  the
thoracic,   abdominal  and  other   regions  was  carried  out.   The  Hvers  were
sub^ectet   to  extensive  hlstologlc,   electron  microscopic  and  biochemical
examlnat'on.  Significant liver  enlargement was  observed  In  male  rats  at  all
dose  lev;ls.   In   addition,  body weights  of  both  male  and  female  rats  were
significantly   reduced.    Electron  microscopy   revealed  an  Increase   In
peroxlsomal  proliferation  at all  dose levels and  an  Increase In  number  of
Tysosome;  at 200 and 1000 mg/kg/day.

    In  tie  study   by  Gray et al.  (1977), male  and  female CO  rats were  fed
dietary  levels  of 0,  0.2,  1 and  2% OEHP  In the  diet  for  17  weeks.   Dally
doses  calculated   from  food  consumption  data corresponded  to  143,  737  and
H40  mg/S,g/day  for males, respectively, and  154,  797 and 1414 mg/kg/day for
females,  respectively.   Body  weight,  food  consumption,  clinical  signs  of
toxlclty,   serum  biochemistry,   urlnalysls  and  hematology  were  monitored.
Gross  and  microscopic  pathologic examinations were performed  on  all  rats  at
the  end of  the study.   Effects were  observed  at  all   levels of  exposure.
Significantly  Increased  absolute and  relative liver  weights  were  observed In
all  expo led groups.  Both males and females fed  either  1 or  2% DEHP had a
significantly  reduced  packed  cell  volume compared with controls.   At  the
0.2%  levd,  liver  weight  was  Increased in both  sexes and  spermatogenesis was
decrease<  in males.

    Carpenter  et  al.   (1953)  conducted  chronic  toxlclty testing  in rats,
guinea  p'gs  and dogs.   Groups  of 32 male and 32 female Sherman rats were fed

04780                               VIII-10                         07/02/91
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 dietary  levels  of 0.04, 0.13 and  0.4% OEHP.   Mean dally Intakes were 20, 60
 and  200  mg/kg/day.   During the  first  year,  male and female rats were housed
 together  until  the  females  became pregnant.  Parental  rats  were maintained
 on  their  respective diets for 2  years.   Offspring of  the rats fed 0.4% DEHP
 were  maintained at  this  dietary  level  for  1  year.  After  1  year,  parental
 rats  were reduced  In  number  to  a maximum of 8/sex/group.   No  effects were
 observed  at   the  0.04  or  0.13%  levels.   At  the 0.4%  level,  decreased body
 weight and  Increased  liver  and  kidney  weights  were observed at  the  end of
 the  first year,  but  no  significant  effects  on  fertility  were  observed.
 While  this  study provides  Information  on  a  chronic NOEL,  the  low survival
 among  controls,  which  experienced  70.3% mortality from  causes  such  as lung
 Infections,  postpartum  complications,  peritonitis, abdominal  abscess  and
 intestinal Intussusception over  the 2-year period, should be noted.
    Guinea  pigs  (23 male  and 23  females/group)  were  also  fed DEHP  In the
diet at  levels  of  0.04 and 0.13%  for  1  year  (Carpenter et a!., 1953).  This
experiment  showed  no  effects  at  the  0.04%  level  (-19  mg/kg/day)  and
Increased  liver  weight at  0.13%  (64  mg/kg/day).   These  results  appear  to
Indicate  that  the  guinea  pig  was  slightly more sensitive to  the  effects  of
DEHP  1n  this  study.   A  group  of  four  dogs  given  capsules 5  times/week
containing  0.03  mt/kg/day  for  19  doses,  then  0.06  ma./kg/day  for  240
doses showed no significant effects (Carpenter et al., 1953).
    Harris  et  al.  (1956)  reported  similar  results  In  a 2-year  rat  study.
Groups of  43  male and 43  female  rats  were fed diets containing  0.1  or  0.5%
DEHF.   No  compound-related  effects  on  mortality  were  observed;  however,
survival over  the 2-year study period  was very low  with 85-95% mortality.
04780
VIII-11
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No effects were  observed  at  the low dietary  level  (-50-80  mg/kg/day),  which
1s consistent with  the findings of  Carpenter  et al. (1953).  At  the  higher
level, focd consumption decreased after  the  first  year.   Increased liver  and
kidney weights were observed In high-dose  rats  sacrificed at 3 and 6 months.
The dally  Intake  at  the  higher dose, calculated for  the  first 6  months,  was
-300-400 rrg/kg/day.  Testlcular atrophy was  not  reported  by  Carpenter  et  al.
(1953) or Harris et al. (1956).

    The oral  NOAEL  for reproductive effects  of  DEHP appears to be  near  the
NOAEL  for  chronic  toxic  effects.   Tomlta  et  al.  (1982a)  reported that  a
single oril  dose of 0.05 ml/kg  administered by gavage  to  mice  on  day 7  of
gestation was associated  with  a decrease  1n body  weight of  viable  fetuses;
however, ro  abnormal   fetuses  were  observed.  Since  the  density   of DEHP  Is
0.985  g/nu,   the  0.05 ml/kg  dose  Is  equivalent   to  49 mg/kg.   Using  the
dose-response curve  for  resorptlons and  deaths,  the authors  calculated  the
NOEL  for   fetal   lethality  to  be   64  mg/kg.   Shlota and  Nlshlmura  (1982)
administered  DEHP  1n   the diets  of  ICR-ICL  mice on days 0-18 of  gestation.
At  the  0.05% level  (70  mg/kg/day)t  the  only  effect  observed was  retarded
ossification.  This  effect  was  thought  to   be   related   to general   and
under-development of  the  fetuses  rather than teratogenlc activity,  since  no
Internal inomolles  were  observed.    At  a  dietary  level  of   0.1%  (190  rag/kg/
day)  the rumber  of resorptlons  and  dead  fetuses  were Increased, although  the
statistical  significance  of  this increase was  marginal   (p=0.05).   At  0.25%
(410  mg/kj/day),  an   Increased  number  of  malformations were  observed  In
addition  lo  Increased resorptlons  and  dead  fetuses, decreased maternal  and
fetal welchts,  and retarded ossification.  At 0.4% and  1.0% (830  and  2200
mg/kg/day}, all  fetuses were dead or resorbed.
04780
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     More  recently DEHP  was  evaluated for developmental  toxldty  In Fischer
 344 rats  and  CO-1 mice  (Tyl  et  al.,  1988).   Dietary  levels of  OEHP  were
 administered  on gestatlonal  days  0-20  to rats  at  0,  0.5, 1.0, 1.5  or  2.0%
 and on  gestatlonal  days  0-17 to  mice  at  0,  0.025,  0.05,  0.10  or  0.15%.
 Corresponding  levels  In  mg/kg/day  were  0,  356,  666, 856, 1054 and 0, 44, 91,
 190,  292  1n rats  and  mice,  respectively.  Tyl  et  al.  (1988)  concluded  that
 DEHP  was  not  teratogenic  at  any  dose tested in  Fischer  344  rats.   However,
 treatment  did  produce .maternal  and other embryofetal  toxldty at  1.0,  1.5
 and 2.0%.   An embryofetal  NOEL  In rats  was  determined to  be   0.5%  (356
 mg/kg/day}.   In mice,  doses  {0.10 and  0.15%)   that  produced maternal  and
 embryofetal  toxldty  also  increased  Incidence  of malformations.   A  dose  of
 0.05%  (91  mg/kg/day)   DEHP   produced  Increased  Incidence of  malformations
 without maternal  or  embryofetal toxldty.   An  embryofetal NOEL In  mice was
 determined to be 0.025% (44 mg/kg/day} OEHP.
    A  study  by  NTP (1984a) tested CD-I mice  using  a  newly developed testing
scheme designated  "Fertility  Assessment by  Continuous  Breeding".   Results of
this  study  were similar  to  those of  Shlota  and Nishlmura  (1982);  however,
the NTP  study focused on  fertility  effects rather  than  teratogenidty.   In
the first phase  of this  test, groups of 20 male and 20  female  mice  were  fed
diets  containing  0.01, 0.1  and  0.3% DEHP  for  7 days  prematlng  and  for  98
days of continuous mating, after which  they were maintained  for 21  days  with
no  treatment.   Daily  Intakes  of  DEHP  were not calculated  by the  authors.
However,   if  one assumes  the  same  dally Intake  rate  as   that calculated  for
the  low-dose CD-I  mice  In   the  cardnogenldty bloassay  by  Kluwe et  al.
(1982a) of 735 mg/kg/day  (averaged for males  and females) for  a 0.3% dietary
level, daily  DEHP Intakes  for  the  lower   dietary  levels of  0.01   and  0.1%
04780
VIII-13
07/02/91
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would be -24 and  243  mg/kg/day,  respectively.   It should be noted that these
calculations do not account  for  differences  1n food consumption due to preg-
nancy, a je  of  mice  or  any  additional  differences between  the  two studies.
At  the 0.3% level, complete  suppression  of  fertility was  observed.   At  the
0.1%  lev«'l, fertility was  decreased  and  various reproductive parameters were
significantly  decreased.   These  parameters  Included  number  of  Utters  per
pair,  anJ  number  of  live pups  per  Utter,  proportion  of  pups  born  alive,
number of male pups born  alive,  live pup weight of females and adjusted live
pup weight  of  males.   A  second  phase of  this  study  used  the  mice  from the
continuous  breeding  phase.   In  this phase  control  ma'les were mated  to  the
0.3%-treeted females  and  control  females  were mated to  0.3%-treated  males.
In addlt'on, control  males were  bred to  control females  to serve as the con-
trol  groi.p  for  the second phase.  Results of  this phase of testing revealed
that  the  decreased  fertility was attributable to effects  of DEHP  In both
males anc females.

    Quantification of Noncarclnoqenlc Effects — DEHP.
    Assessment  of Acute  Exposure  Data  and  Derivation  of  1-day  HA -- Liver
enlargemtnt,  testlcular  atrophy  In  males,  depressed  weight gain  and death
have  all  been  observed  after oral administration  of  single doses of  DEHP to
rats.  LI)   s  for  DEHP have  been measured  in a variety  of  species  and range
from  26  g/kg  (rats)  to  34 g/kg (rabbits).  In  rats, neonates  and sucklings
are  more  sensitive to the weight  gain and  lethal effects  of  DEHP  than  are
adults.  Oostal et  al.  (1987a)  administered  five successive doses (gavage in
corn  oil)  of   0,  10,  100,  1000  or  2000  mg/kg/day  to  six groups  (9-10
pups/groip) of  rats, 6-86  days  old.   For  neonates and  sucklings,  doses of
2000  mg/kg/day  were  lethal  and  1000 mg/kg/day caused depressed  weight gain
04780
VIII-14
05/16/91
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 1n all groups and  Increased  mortality  1n  sucklings 14 days old.  Adults  (86
 days   old)   were   less  sensitive  to  these  effects,   with no   Increase  In
 mortality  at  any  dose  and  effects on  weight  observed  only  at  the  2000
 mg/kg/day  dose  level.   At   100  mg/kg/day,  sucklings  and  adults  exhibited
 Increased  I1ver-to-body weight  ratios.
     Effects  of  acute  oral  exposure to D£HP on the liver have been studied by
Mitchell  et  al.  (1985)  and Mangham  et  al.  (1981).   Mitchell  et  al.  (1985)
administered  DEHP  1n  the diet of  rats  (4/sex/group)  at nominal doses  of 50,
200  and  1000 mg/kg/day.   H1stopatholog1c,  biochemical  cytogenetlc  analyses
we-e  conducted  on days   3,  7,  14  and  28,  and  at  9  months  of  dosing.
Indications  of  hepatotoxldty   (Increased   liver  weight,   decreased  hepatic
gljcose-6-phosphatase activity)  were  first  observed  1n  the 50 mg/kg/day dose
males  at 14  days  of  treatment.  Mangham  et  al. (1981)  observed decreased
teitlcular weight, microscopic  changes  1n the seminiferous tubules,  enlarged
11*er,  decreased  activity  of   sucdnate  dehydrogenase  and  decreased  body
weight after seven doses of 2500 mg/kg/day.
    In  addition,   DEHP  can  cause  reproductive  and developmental  toxlclty.
Tyl et  al.  (1988)  observed fetotoxlclty In mice and rats  at  doses  of  91  and
666  mg/kg/day,  respectively.   Mice  were  dosed on days  0-17  of  gestation
while   rats   were   dosed  on   days   0-20.    NOAELs  for   reproductive   and
developmental effects  of 44  mg/kg/day  (mice) or  357  mg/kg/day  (rats)  were
Identified.   These  observations are  supported by  the  work of TomHa  et  al.
(1982a).   Mice  were  administered  single  doses   of   50  »l  DEHP/kg  (-49
mg/kg)  or  100  vl  DEHP/kg  (-99  mg/kg)   to pregnant  dams  at  day  7  of
gestation and decreased fetal weights at birth were observed.
04780
VIII-15
05/16/91
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    For exposure of  5  days  or less, developmental  effects  In  mice and liver
enlargement In rats  are  the most sensitive endpolnts  of  toxUHy.  From the
studies described previously, 1t  Is not  possible  to determine  whether  a dose
of  44  mc/kg/day,  the  NOAEl  for developmental  effects,  would  cause  liver
effects 1n rats, observed at  100  but not 10 mg/kg/day. Therefore, the NOAEL
for  live-  enlargement  of  10  mg/kg/day  from  Dostal et  al.  {1987a}  was
selected as the basis for the 1-day  HA  for DEHP,  derived as  follows:
        HA
                               Omq/kq/day x 10 kq
                                 100 x 1  I/day

where:
    10 mg/kg/day

    10 kg
    100

    1 l/day
=   NOAEL  based  on lack of liver enlargement (Dostal et
   al., 1987a)
=  assumed weight of  a  child
=  uncertainty   factor,   according   to   U.S.   EPA  and
   ODW/NAS  guidelines  for use  with a  NOAEL  From an
   animal study
=  assumed water  consumption by a child
    Assessment of Short-Term Exposure Data and Derivation  of  10-day  HA —
Effects en  the  liver  appear  to be  the  most  sensitive endpolnt of  toxlclty
for 10-dey exposure.   Mitchell  et  al.  (1985)  observed liver  effects  in  rats
after 14  days  at doses as low  as  50 mg/kg/day,  a dose similar to  the NOAEL
for developmental toxldty In mice  (44 mg/kg/day}.   A  NOAEL  for  the observed
liver  effects  {decreased glucose-6-phosphatase  activity,  Increase  In liver
weight,  increase In  hepatocyte  I1p1d   content)  was  not  Identified.   This
number  1:; consistent  with  the 1-day  HA  If  1t  were  adjusted  for  a  10-day
exposure  period.  However,  the Mitchell   et al.  (1985) study examined other
organs  1r  addition  to the liver and provides a  better estimate of  a 10-day
04780
                    VIII-16
07/02/91
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 exposure.   The  10-day  HA  1s  also  protective  of  developmental  toxldty
 Identified  at  91.07  mg/kg/day  In  mice  and  666.39  mg/kg/day In rats by Tyl  et
 al.  (1988).   Thus,  the Mitchell et  al.  study  Is  chosen  to derive the 10-day
 HA as  follows:
                  10.day HA s50mq/kq/daY x 10 kq  =  Q^
                                1000  x  1  a/day
where:
    50 mg/kg/day = LOAEL  based  on liver  enlargement  (Mitchell et  al.,
                   1985)
    10 kg
    1 l/day
assumed weight of a child
    1000         = uncertainty   factor,   according   to  U.S.   EPA   and
                   OOW/NAS  guidelines  for  use  with  a  LOAEL  from  an
                   animal study
assumed water consumption by a child
    Derivation  of  Longer-term   HA  —  Subchronlc   oral   studies   have  been
conducted with DEHP, however none  of  the  studies  Identify a NOAEL.   Mitchell
et  al.   (1985)   observed   significant   liver   enlargement  In  male  rats
administered  50,  200  or 1000 mg/kg/day  for  9 months.  No  clear  progression
of hepatotoxU effects  was  observed  from 3-, 7-,  14-  or  28-day  time points.
This 1s  supported by  the subchronlc  study by Gray  et  al.  (1977)  where liver
weights  were  Increased  In both  sexes  of  rats  and  spermatogenesls  was
decreased  1n  males  (143 mg/kg/day dose  level  In  males;  154 mg/kg/day  dose
level  1n females).
    Deriving  the  longer-term  HA  based  on  the  LOAEL  of  50  mg/kg/day  Is
protective of  the  reproductive (NTP,  1984a)  and developmental  toxldty  (Tyl
et al.,  1988)  observed  1n  mice at doses  of 243  and 91 mg/kg/day,  respec-
tively.   The  reproductive  study  by  NTP  (1984a)   showed  no  effects  on
Q4780
                  VIII-17
07/02/91
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fertility In mke  fed  0.01% (24 mg/kg/day) In the diet  for  7  days  prematlng


and 98  cays continuous  breeding.   The  next  highest  dietary  level of  O.TX


(240 mg/>g/day) significantly reduced fertility.  Tyl  et al.  (1988)  observed


fetotoxUHy In mice and rats at 91 and  666 mg/kg/day.   NOAELs for  reproduc-


tive and  developmental effects  of 44  (mice)  or 357  (rats) mg/kg/day  were


Identified.




    Therefore,  based on  a  LOAEL for  hepatotoxlclty  In rats the  longer-term


HA values are calculated as follows:
                                50 mq/kq/day x  10  kg
               Longer-term HA =                      =  0.5  mg/l,
                                  1000 x 1  l/day
                  (child)
where:
    50 mc/kg/day « LOAEL based  on  liver enlargement  (Mitchell  et al.,
                   1985)
    10 kc
    1 l/cay
= assumed weight of a child
    1000         = uncertainty  factor,  according   to   U.S.   EPA  and
                   ODW/NAS  guidelines  for  use  with a  LOAEL  from  an
                   animal study
  assumed water consumption  by  a  child
                           50 mq/kq/day x 70 kg
          Longer-term  HA  =                      = 1.75 mg/l
                            1000  x 2 I/day
              (adult)                             (rounded to 2 mg/l)
where:
    50 mc/kg/day = LOAEL based  on  liver enlargement  (Mitchell  et al.,
                   1985)
    70 kc
    2 t/cay
= assumed weight of an adult
    1000         = uncertainty factor, according to U.S EPA and ODW/NAS
                   guidelines for use with a LOAEL  from an  animal  study
  assumed water consumption by an adult
04780
                    VIII-18
07/02/91
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    Assessment  of Long-Term  Exposure Data  and  Derivation  of a  DHEL — No
data  were  available  on  the  effects of  chronic  human  exposure  to  DEHP.
Carpenter  et  al.  (1953) reported  no  effects  1n rats exposed  for 2  years to
dietary  levels of  0.04%  and  0.13%,  equivalent  to -20  and 60  mg/kg/day,
respectively.  There was  low  survival of both  control  and treated  rats dur-
ing the  second year.  However,  results  with  other  species  and  studies sup-
po*t  these results.   Carpenter et  al.  (1953)  also  reported  that  no effects
we-e  observed  1n  guinea  pigs exposed to 0.04% {-19 mg/kg/day) 1n  the diet
for 1 year; however, Increased  liver  weights were reported.   Increased liver
weight was also  observed  In  guinea  pigs  exposed  to  0.13%  DEHP  (64  mg/kg/
day).   No  hlstologU  effects  on  liver  tissue  were  observed.   The  suggested
LOAEL Is  -19  mg/kg/day.   A study  by  Harris  et al.  (1956)  showed  no effects
In rats fed 0.1%  DEHP  (50-80  mg/kg/day)  for  2  years.  Again, low survival In
all  groups places  limitations  on Interpretation  of  the  results  of  this
study.  A  recent  reproductive  study  showed  no effects on  fertility  in mice
fed 0.01% DEHP  1n   the  diet   for  7  days  prematlng and  98  days  continuous
breeding   (NTP,  1984a).    Assuming food  consumption  was  similar  to  that
reported by Kluwe et al.  (1982a)  for  mice,  dally Intake was calculated to be
24 mg/kg/day.  The next  highest dietary level   of  0.1%  significantly reduced
fertility.  The approximate  dally  Intake for  this dietary level was  calcu-
lated to  be  240 mg/kg/day.   Also,  Shlota and  NUhlmura  (1982) reported only
reduced ossification In  mouse fetuses born  to  dams  fed  70 mg/kg/day on days
0-18  of  gestation.   Tyl  et al.  (1988)  determined an embryofetal NOEL  to be
356.74 mg/kg/day  In  Fischer   344  rats  and  44.07 mg/kg/day  1n  CD-I  mice.   In
light of  these results and the  fact that  no  NOAEl was  Identified,  lower than
the lowest LOAEL  observed, -19  mg/kg/day reported by Carpenter et  al.  (1953)
was selected for use In calculating the DUEL  (U.S. EPA,  1991).
04780
VIII-19
07/02/91
-------
    Uslnc this IOAEL, the DWEL would be derived as follows:
where:
                     19 mg/kg/day
               RfD • 	—	  = 0.019 mg/kg/day
                         1000                 y
                                    (rounded to 0.02 mg/kg/day)
    19 mc/kg/day = LOAEL  derived  from  oral  exposure  to  guinea  pigs
                   (Carpenter et al., 1953)
1000
                 « uncertainty  factor,  according   to   U.S.  EPA  and
                   OOW/NAS  guidelines  for   use  with  a  LOAEL  from  a
                   subchronlc animal study
                  DWEL
                                 2 I/day

where:
    0.02 mg/kg/day  = RfO
    70 kc           = assumed weight of an adult
    2 l/cay         * assumed water consumption by an adult

    The  -day  HA,  10-day HA,  longer-term  HA and DWEL  values  calculated  for
DEHP  and  the  effect levels  used  In the derivations are  summarized  In  Table
VIII-1.
    Studies   Considered   for   Noncardnoqenlc   Quantification
                                                                     B8P.
ToxicHy  of  BBP  Is  limited to  a  few studies.   The most  commonly  observed
effects 
-------










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VIII-21
07/31/91
-------
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04780
                                                       VIII-22
                                                                                                      07/31/91
-------
liver  and kidney  weights  were  significantly Increased.   In  addition,  the
Incidence  of  proximal  tabular  regeneration  of  the  kidney 1ncr-:^sed  In a
dose-related manner beginning at the 0.625% dose level.  At  the  2.5% and  5.0%
levels,  effects  Included  decreased  body  weight,  slightly  decreased  food
consumption, decreased  weights  of  testes,  epldldymus,  seminal vesicles  and
thymus, hlstologlc atrophy of testes and  accessory  sex  organs,  and decreased
bone  marrow cellularlty.   Dally  Intakes  of BBP  were  not  calcu-   lated.
However,  dally  food  consumption  and  body weight  were  estimated  from  the
figures presented by Agarwal et al,  (1985a).   At  the  lowest dietary  level of
OJ>25%, rats weighing' -250 g consumed 15  g  of food per  day or 60 g  food/kg
bw/day.   The  dally  Intake  of  BBP  was calculated  to  be 375  mg/kg  bw/day.
Since body weights and  food consumption were  not  affected at 1.25%,  the same
food  consumption  and  body   weight  were   used  and  the  dally  intake  was
calculated  to   750  mg/kg  bw/day.    Food  consumption  and  body  weight  were
decreased  at  2.5  and  5.0%,  respectively.   Again,  approximating  from  the
figures  In  Agarwal  et  al.  (1985a), 200 g  rats  at  the  2.5% level  consuming
10 g  food/day and  150 g rats at  the 5.0% level consuming  5 g  food/day, the
daily Intake was 1250 and 1667 mg/kg bw/day, respectively.
    Lake  et  al.   (1978)  administered  160,  480  or  1600  mg/kg/day  BBP  by
gastric  intubation  for 14  days  to  six  male Sprague-Dawley  rats  per  group.
Biochemical  or  morphologic  changes  In  the liver were  not  observed  at  160
mg/kg/day.   Activities  of  ethylmorphlne  N-demethylase and cytochrome oxldase
were significantly  Increased at  the  480  and 1600  mg/kg/day BBP.  Significant
liver   enlargement  was   observed  at   1600  mg/kg/day  in   addition   to
ultrastructural  changes,  such  as gross  dilation  of the  rough endoplasmlc
04780
VIII-23
07/02/91
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retlculum  and  Increased number  of peroxlsomes.   Effects  on testes  weights
were  not  observed  1n  the  160  or  480  mg/kg/day  animals;  however,   1600
mg/kg/day  BBP  produced  marked  depression  of  both  absolute  and  relative
testes weights as well as  severe  testlcular  atrophy.  Testlcular  atrophy was
observed  in 1/3 animals administered 480 mg/kg/day.

    A  second  study was conducted  to confirm  the testlcular effects.   Both
Sprague-Dawley  and  Mlstar  Albino  rats  were   treated  with  480  and  1600
mg/kg/day  BBP  for  14 days.   A  significant  depression 1n  either  absolute or
relative  liver  and  testes  weight was  observed  In both  strains  of  rats  at
1600  mg/l;g/day  BBP.  Hlstologlc  examination revealed testlcular  atrophy In
both  strains  (1600  mg/kg/day)  with  the extent of  the  lesions  being  more
severe  1i,  the Sprague-Oawley  strain.  At  480 mg/kg/day  BBP, 1/6 had testlcu-
lar atrophy, whereas the Wlstar albino strain revealed no hlstologlc changes.
          are  few  oral  long-term BBP studies.   In a final report, NTP (1985)
conductei  toxlclty  and  mating  trial  studies  1n  F344  rats.   The  toxlrlty
portion  rfas  conducted  as  a dose range-finding study to establish a no effect
level  and   the   dose   response  curve  for  BBP.   Rats  were  administered
concentrations of  either  0,  0.03,  0.09.  0.28,  0.83 or 2.50%  (0, 17, 51,  159,
470  and  1417  mg/kg/day)   BBP  1n  the  diet  for  26 weeks.  Powdered  BBP was
mixed  1r  to standard  rodent meal  diet.   Because  of  the  manner 1n which the
BBP  was  administered considerable  waste  and spillage was found  especially at
the  highest  dose level.   Therefore, the  dose  conversion for  the highest was
based  on a  554 food  consumption  rate/mg  rat body weight.  There were 15  male
animals  in   each  dose  group,  starting at  6  weeks of age.   Throughout the
04780
VIII-24
                                                                      07/02/91
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study,  body  weight gain  was  significantly depressed  at  the 2.5% BBP  level
when  compared  with  the  controls.  There  were no  deaths  attributed to  BBP
toxlclty.  All  the  rats  given 2.5% BBP had small  testes upon  gross  necropsy
at  the  26-week  termination.   Five of  11  had  soft testes, and  1/11  had  a
small  prostate  and  seminal   vesicle.   At   0.83%,  significantly  (p<0.05)
Increased absolute  liver  weight,  llver-to-body  weight,  Hver-to-braln  weight
ratios  and   Increases  In mean  corpuscular hemoglobin  were noted.   In  the
0.03, 0.09,  0.28  and  0.83% BBP dose groups there  were no  grossly observable
effects on male reproductive  organs.   The  kidneys  of  six animals  1n  the 2.5%
grcup contained  focal' cortical  areas  of 1nfarct-l1ke  atrophy.   In addition,
testlcular lesions  were  also  observed  at the 2.5%  dose  level.   Lesions  were
characterized  by  atrophy  of  seminiferous  tubules  and aspermla.  The  other
treatment groups  showed  no  evidence  of  abnormal  morphology  In any  other
organs.

    Hlstopathologlc changes  were also  seen  at the 2.5% BBP  level  after 10
weeks of  exposure 1n the  mating  trial portion of  this  study.   After  hlsto-
pathologlc examination,  testlcular  lesions were characterized  by atrophy of
seminiferous tubules  and a near  total absence  of mature  sperm  production.
When  10/30  females  successfully  mated  with  the 2.5%  treatment  level  males,
none  were pregnant  at necropsy.   The  Investigators concluded  that  the  data
suggest a depression  In  male  reproductive organ weights  by either  a direct
or Indirect  toxic effect  after  2.5% BBP administration.   BBP at  0.83% 1n the
diet did not result 1n any treatment-related effects.

    The  only other  Information  on  the subchronlc  effects  of BBP  Is taken
from an unpublished  study by  Monsanto  (1972).   Rats fed diets containing BBP


04780                                VIII-25                         07/02/91
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at  levels of  0.25  {125  mg/kg/day)  and  0.50%  (250 nig/kg/day}  for  90  days
showed  no  toxic effects.   A  dietary  level  of  1.0%  {500  mg/kg/day)  BBP
resulted  1i  Increased  liver  weight.   Levels of 1.5  (750  mg/kg/day)  and 2.0%
{1000  mg/lig/day) BBP  were  associated  with  Increased  liver  weight  and  a
decrease  1i  growth  rate.   No effects were  observed  In  dogs  administered BBP
1n  capsules  at  levels equal  to  1.0, 2.0 and  5.0% of  the diet.   No further
details of this  study were available for review.

    Quantl Mcatlon of Noncarclnogenlc Effects --  BBP.
    Assessment of Acute  Exposure Data  and  Derivation  of the  1-day  HA — No
Information was  available on  the  effects of BBP  In humans.   The only studies
available  >n acute oral toxldty  In  animals used  lethality  as  the toxic end-
point  or  were  Inadequate  for  deriving a 1-day  HA.    Therefore,  lack  of
sufficient  data  preclude  the  derivation  of  a  1-day  HA  for  BBP.   It  Is
recommended  that the  10-day  HA  of  20 mg/i  be   adopted  as  a  conservative
estimate  f>>r the 1-day HA.

    Assessnent  of   Short-Term  Exposure  Data  and  Derivation  of  a  10-day
HA —   Information  presented  In a  14-day  study  was used to approximate the
10-day HA  values.  Agarwal et  al.  (1985a)  administered  BBP  to  male F344 rats
In  the  dltt  for  14 consecutive days  at dose levels of 0.625,  1.25,  2.5 and
5.0%.   Effects  observed  beginning  at   the  0.625% level were significantly
Increased  liver  and  kidney   weights.   Dose-related hUtopathologlc  changes
{proximal  tubular regeneration)  were also  noted  1n the  kidney  beginning at
the 0.625% level.  Using  approximations of  food  consumptions  and body weight
obtained  from  figures presented  In  this study,  the dally  Intake at  0.625%
level was  :alculated to be 375 mg/kg/day.
04780
VII1-26
07/31/91
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    In male.,Sprague-Dawley rats  administered  160,  480 or 1600 mg/kg/day 8BP
for 14 days by gastric  intubation, biochemical or morphologic changes  In the
liver  as  well as  effects on  testes  weights  were  not  observed  1n  the 160
mg/kg/day  dose  group   (Lake  et  a!.,  1978).   However,  at 480  mg/kg/day
activities  of  ethyl   morphine   N-demethylase  and  cytochrome  oxldase were
significantly  Increased   and   testlcular  atrophy   was  observed  in  1/3
Sprague-Dawley rats 1n  the  first portion  of this experiment.   In  the  second
portion,    the   480  mg/kg/day   dose   Induced   testlcular  atrophy  1n  1/6
Sprague-Dawley  rats,   whereas  the  Wlstar  albino  strain revealed  no  such
effects (Lake et  a!.,  >978).

    When  comparing the  two  studies  Lake et al.  (1978)  Identifies  a  NOAEL  of
160 mg/kg/day.  It Is  questionable whether  480 mg/kg/day represents  a  NOAEL;
however,  Agarwal et al.  (1985a)  observed  significant  Increases  In  liver  and
kidney weights  and kidney  pathology at  375 mg/kg/day,  which   represents  a
LOAEL.   It  Is  therefore  recommended  that  the  NOAEL  of  160  mg/kg/day
Identified  In  the  Lake et al.,  (1978}  study be used  1n  deriving  the  10-day
HA.  Although the method  of  treatment was  gavage in the study by Lake et al.
(19"?8) and  diet  In  the study by  Agarwal  et al.  (1985a),  treatment-related
effects across similar  dose  ranges,  Including Hver  effects  1n both studies
In  two sensitive  strains of rats,  support  use of 160  mg/kg/day  as  NOAEL  In
rat> given BBP orally for 14 days.

    The 10-day HA 1s calculated as follows:
       10-day
160 mq/kq/day x 10 kg
    100 x  1  i/day
= 16 mg/l (rounded to 20 mg/l)
04780
                   VIII-27
                                                                      07/31/91
-------
where:
    160 mt|/kg/day
    10 kg
    100

    1 l/diy
NOAEL based on  the  absence  of liver and testkular
effects from animal  data (Lake et a!.,  1978}
assumed weight of a  child
uncertainty  factor,  according  to  U.S.  EPA  and
ODW/NAS  guidelines  for  use  with  a NOAEL  from  an
animal study
assumed water  consumption by a child
    Assessment  of   Longer-term  HA — Long-term  exposure  to   BBP   causes
adverse  (ffects  to  the  testes  of male  rats.   The  only study available  for
                      t
the derivation of  longer-term HAs 1s the 26-week feeding  study  conducted  by
NTP {1985).   Male  F344  rats consuming a dietary  level  of  2.5% BBP exhibited
testUular  lesions  characterized by  atrophy  of seminiferous  tubules  and
aspermla.   The  corresponding  dose   from   data  given,   assuming  5%  food
consumption/day and  200 g  body  weight, Is  1417 mg/kg/day.   At  this  level
rats  also  experienced   significantly  depressed  body   weight   gains   and
significant Increases In the  organ-to-body weight ratios  In  the  brain, right
kidney, light testes  and liver.   Rats  given  dietary  levels of  0, 0.03, 0.09,
0.28 and  0.83V.  BBP  for  26  weeks  exhibited  no grossly  observable  effects  on
male reproductive  organs.   Corresponding doses assuming -300 g  bw and -17 g
of food  :onsumpt1on/day  from  data presented  1n  the report  are  0, 17.0, 51.0,
159 and  470  mg/kg/day,  respectively.   At  0.83%,   the effects   noted  were
significantly   (p<0.05)   Increased   absolute   liver    weight.   Increased
Hver-to-body weight  and Hver-to-braln weight ratios  and  Increases  1n  mean
corpuscular hemoglobin.   Liver-to-body  weight ratios significantly (p<0.05)
Increased for the  brain,  right  kidney and liver  at  the 2.5% level; however,
llver-to-braln weight  ratios  did not  significantly   (p<0.05)  Increase.   The
                                     it T T T  -"Id
-------
 differences  may  have  been  due  to  the  reduced  weight  gain  and  testlcular
 effects  at 2.5%  BBP.   The liver may be  a  more  sensitive endpolnt  than  the
 testes  since  liver  effects  were  observed  at  a lower  level  (0.83%)  than
 testlcular  effects  (2.5%).  Therefore,  0.28%  or  159 mg/kg/day  will  be  used
 as a NOAEL  to derive the longer-term HAs as follows:
              Longer-term HA
                  (child)
                  159 mg/kq/day x 10 kg   ,, Q    ,
                  	inn/x i o/da'v—  -15.9 mg/8,
                       luu x  i i/oay    (rounded  to 20 mg/4)
where:
    159 mg/kg/day = NOAEL  based  on  the  absence  of  Increased  liver
                    weights In rats (NTP, 1985)
    10 kg
    100

    1 i/day
    = assumed weight of a child
    = uncertainty   factor,   according   to   U.S.   EPA  and
      ODW/NAS  guidelines  for  use  with a  NOAEL  from an
      animal study
    = assumed water consumption by a child
Longer -ter. HA - 159 .qAq/day x 70 kq
  (adult)                tuu
                                                     ^
                                          x * l/ady   (rounded to 60 mg/i)
where:
    159 mg/kg/day
    70 kg
    100

    2 i/day
    = NOAEL  based  on   the  absence  of   Increased   liver
      weight In rats  (NTP, 1985)
    = assumed weight  of  an adult
    = uncertainty   factor,   according   to  U.S.   EPA  and
      ODW/NAS  guidelines  for  use  with  a NOAEL  from  an
      animal study
    = assumed water consumption by an adult
04760
                        VIII-29
07/31/91
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    Assessment  of Long-Term  Exposure  Data and  Derivation of  a  DUEL -- NTP
 (1985)  Is  also  the  only  available  study  for  the derivation of the DWEl (U.S.
 EPA,  1991).  The  DWEL Is derived as follows:
    Step  1 - RfD Derivation
                  RfQ
                            uuu
                                      ^ Q<159  mg/kg/day
                                     (rounded 0,2 mg/kg/day)
where:
    159 mg/kg/day = NOAEL derived from orally exposed rats (NTP, 1985)
    1000
                   uncertainty  factor,  according   to   U.S.  EPA  and
                   OOW/NAS guidelines  for  use wHh a NOAEL  from  animal
                   data, for  less  than  lifetime  exposure  and to protect
                   sensitive members of the human population
    Step 2 - DWEL Derivation
                     DWEL  =  —
                                               kq
                                 2 i/day

where:
    0.2 mg/kg/day = RfD
    70 kg
    2 l/day
                      assumed weight of an adult
                      assumed water consumption by an adult
    The  1-day HA,  10-day  HA  and  DWEL  values  calculated  for  BBP and  the
effects levels used In calculation are summarized 1n Table VIII-1.

    Studies   Considered   for   Noncardnogenlc   Quantification  —  DBP.    No
Informatlcn was  found 1n the  available  literature on the effects  of  DBP In
humans anl  Information on  effects  in animals  1s  limited.   The teratogenlc
effects o:  PAEs  following  oral  administration  were studied  by  Nlkonorow et
04780
                                     VIII-30
07/31/91
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al.  (1973).   In  this  study female WUtar rats were administered  120 and  600
rag/kg/day  DBP   In  olive   oil   for   -3   months   and   during   mating.   Upon
confirmation  of  conception the  administration of DBP was discontinued.   On
day  21  the  uteri and  fetuses were removed.   Results  of  this  study  Indicated
that  fetal  weight  was significantly  (p<0.05)  reduced at 600  mg/kg/day DBP.
No   detectable   differences  were   observed  In   the  number   of   sternum
ossification  foci,  the  development  of  the bones at  the base of the  skull,
pa**  on  the  front  and hind legs, or  rib  fusion  1n  fetuses  from  treated rats
at either dose level when compared with the control  animals.
    Cater et  al.  (1977) found  that  DBP Induced testlc'ular  atrophy  In  young
{3-4 weeks old) male Sprague-Dawley  rats.   DBP  was  dissolved In corn on and
administered by gavage  In  doses  of  500,  1000 and 2000 mg/kg/day for  14  days,
while  control  animals  received  corn  oil  In a  volume  of  5 ml/kg.   Testes
we'ghts were measured on days  4  and  6  for  500,  1000 and 2000 mg/kg/day  doses
of  OBP.   In  addition  body weight  and relative  liver,  kidney and  testes
weights  were  measured  on  days  3,  7,  10  and  14  at  2000 mg/kg/day.   The
Initial  effect  was  a  progressive  reduction  In  weight  of the  testes.   At  4
days, however, 500 mg/kg/day DBP did not have an effect on testes weight.  A
significant (p<0.05) reduction  1n  the  relative  testes weight occurred within
6 days at  500 mg/kg/day and within  4  days  at 1000  (significance p<0.01) and
2000  (significance  p<0.001) mg/kg/day.  By  14  days,  the reduction  at  2000
mg/kg/day amounted to 60-70% of  the  original weight.   Since there was also a
decrease 1n body weight,  the authors used  "relative testes weight" and  found
that  on  this basis  there  was  still  a  significant  loss  of  testes  weight.
There  was   a  nonstatlstlcally  significant   Increase   In  liver  weights.
Hlstopathologlc examination of  testes  tissue after  4  days  of  2000 mg/kg OBP
exposure revealed a diminution of both  spermatocytes and spermatogonla.
04780
VIII-31
07/02/91
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    In a  detary  study  DBP  was  fed to male and female Fischer 344 rats at 0,
0.6,  1.2  Jnd 2.5%  for  21  days  (CMA, 1986).   Corresponding  dose  levels  were
0,  624,   1234  and  2156  mg/kg/day  for  males  and  0,  632,   1261,  and  2107
mg/kg/day   for   females.    Absolute   and  relative   liver  weights   were
slgnlfIcartly  Increased  In both  male  and  female  rats  at  all  treatment
levels.   4ale  rats fed 2.5% DBP  had severe  testlcular atrophy  and  signifi-
cantly  lower testes  weight.   Samples  of  liver  from rats  administered  the
2.5%  level  showed  "moderate" peroxlsomal  proliferation.   In addition  lauMc
acid  11-  and 12-hydroxylase Increased  In  males given 0.6,  1.2 and  2.5% and
In   females  given   2.5%.    Cyanide-Insensitive  palnvUoyl   CoA   oxidation
Increased at 1.2 and 2.5% In males and 2.5% In females.

    Smith  (1953)  studied the  effects of  feeding DBP to  groups  of 10  male
5-week-old  Sprague-Dawley  rats,  weighing  55-65 g.   Rats  were  fed  dietary
levels of 0, 0.01,  0.05,  0.25 and  1.25% DBP for 1 year.   The dietary Intakes
for DBP vere 0, 5,  25,  125  and  600 mg/kg/day, respectively,  estimated from a
figure depicting dally  Intake In mg/kg  In  Smith (1953).   Survival  rates were
not repotted for  the  three  lowest dose  groups.   In  the group fed  1.25% DBP,
half  (presumably  5/10) of  the  animals  died  during  the   first  week  of  the
experlmert  while   the  remaining   animals  gained   weight  comparable  with
controls    It  was not  Indicated  whether  the deaths were  thought  to  be
treatmen .-related.  Necropsies were  performed  when  rats showed marked weight
loss  or signs  of  severe Infection.   Animals alive  at  the  end of  1  year were
sacrlf1c;d  and examined for gross  pathologic  changes.  While 1t  was  stated
that  several  organs were sectioned  and stained,  the results of  hlstologlc
evaluatljn  were not reported.   Of  the animals  surviving,  no adverse effects
on growth,  survival, gross  pathology or  hematology  were observed among those
04780
VIII-32
07/02/91
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fed diets  containing  0.01,,  0.05  or  0.25% DSP.  The dally Intake of food and
plastlclzer  (mg/kg  bw/day)  decreased  as  the rats  Increased  1n  size.   No
changes  In hematologk  parameters or  gross  pathology were  observed  at  any
dose level.
    Shlota  and  Nlshlmura  (1982)  found  retarded  ossification  1n  mice  fed
diets of 80, 180, 370, 660 and  2100  mg/kg/day  DBF  on  days  0-18 of  gestation.
At  the  660  mg/kg/day level, reduced  fetal  weight  and retarded  ossification
were observed.  Among  rats  fed  diets  of  2100  mg/kg/day,  decreased  maternal
weight  was  observed  along  with  reduced  weight  In  the  fetuses,  retarded
ossification and neural  tube  defects In  the fetuses.   The authors  concluded
that delayed ossification was related  to the general  underdevelopment  of  the
fetuses.  The maximum  nonembryotoxlc  dose  as  stated by  the  authors  would be
370 mg/kg/day D8P.
    Quantification of Noncardnogenlc Effects — DBP.
    Assessment  of  Acute  Exposure  Data and  Derivation of  the  1-Day HA  --
No  information  was  found In  the  available  literature on  the acute  toxldty
of  DBP  to  humans.   Cater et  al.  (1977)  found  that  DBP  Induced  testlcular
atrophy  1n  young  (3-4  weeks  old)  male  Sprague-Oawley  rats.    DBP  was
administered  by gavage  In   doses  of  500,  1000  and   2000  mg/kg/day for  14
days.   Effects  of treatment  on  body  weight  and relative  liver,  kidney  and
testes weights  were  measured on  days  3, 7,  10  and  14  at  2000  mg/kg/day.   In
addition,  testes  weights were measured on  days  4  and 6  for  500,  1000  and
again  for   2000 mg/kg/day.    At  4  days of  500  mg/kg/day  treatment  testes
weights  were   not   affected.    Liver  weights   Increased  but  were  not
statistically significant.   Treatment  at  4  days  of 1000  and  2000 mg/kg/day
04760
VIII-33
05/16/91
-------
 significantly  reduced testes  weight  and  at  2000 rog/kg/day  diminished  both
 spermatocytes  and  spermatogonla.   By  14  days   2000  mg/kg/day  reduced  the
 testes  to 60-70% of the original weight.

     In  another  study, CHA (1986) reported  significant  Increases  In absolute
 and  relative  liver  weights after  21  days  of exposure to doses of 624 and 632
 mg/kg/day In male and female Fischer 344 rats, respectively.

     Cater et  al.  (1977)  Identified  a  NOAEL of  500  mg/kg/day for testlcular
 effects  in  male rats.   Liver  weights  Increased but were  not  statistically
 s1gn1f1cait.   After 21  days  of  exposure, CMA  (1986)  reported  significantly
 Increased absolute  and  relative  liver  weights.   It  appears  that  for a 1-day
 exposure  testes  are  the  most  sensitive  organ  and,  therefore, the  NOAEL of
 500  mg/kg'day will  be used to derive the 1-day HA as follows:
                        HA = 500 mq/kq/day x 10 kq ^
                                100 x 1 I/day
where:
    500 mg/kg/day = NOAEL  based  on  the  absence  of  decreased  testes
                    weight from animal data (Cater et al., 1977)
    10 kg
    100

    1 I/day
= assumed weight of a child
= uncertainty  factor,  according  to  U.S.   EPA   and
  OOW/NAS  guidelines  for  use  with  a  NOAEL from  an
  animal study
= assumed water consumption by a child
    Assessment  of  Acute Exposure  Data and  Derivation  of the  10-Day  HA —
Cater  et  al.  (1977)  observed  decreased  spermatocytes and  spermatogonla as
well  as  significantly reduced  testes weight  after  4 days  exposure to 2000
mg/kg/da>  DBP.   In addition,   testes  weights  were  significantly  reduced at
04780
                   VIII-34
07/02/91
-------
 500  mg/kg/day  at 6  days  and  1000 mg/kg/day at  4  days of exposure  (LOAEL  =
 500  mg/kg/day).   CMA  (1986)  reported  significant  Increases  In absolute  and
 relative  liver weights  after  21 days of exposure  to  -600  mg/kg/day  In  rats.
 However,  Smith  (1953)  reported  a  NOAEL  of  125  mg/kg/day  for   growth,
 survival, gross  pathology or  hematology after  1  year  exposure  of  08P to male
 rats.   In light  of  these  data,  125 mg/kg/day  appears to  be a  reasonable
 estimate  of a NOAEL  after 10  days  of exposure  and  will be  used to derive the
 10-day HA In addition to the  longer-term HAs and DWEL.

    The 10-day HA Is calculated as follows:
                  10-day
         125 mq/kq/day x 10 kg
              100  x  1  i/day
12.5 mg/s.
(rounded to 10 mg/i)
where:
    125 mg/kg/day = NOAEL  based  on the  absence of  Increased  mortality
                    and hematologlc effects (Smith, 1953)
    10 kg
    100

    1 l/day
= assumed weight of a child
= uncertainty  factor,  according  to  U.S.  EPA   and
  ODW/NAS  guidelines  for  use with  a  NOAEL  from  an
  animal study
= assumed water consumption by a  child
    Derivation  of   Lonqer-Term   HA  —  The   only   study  available  for  the
derivation  of  longer-term  HAs  Is  by  Smith  (1953).   Male  rats  fed  diets
containing  5.0,  25  and  125  mg/kg/day  for  1  year  experienced  no  adverse
effects on  growth,  survival,  gross  pathology or  hematology.   At a level of
600 mg/kg/day  DBP,  half of  the  animals died.  The  remaining animals gained
weight  as  did  the  controls.  The limitations of  this  study,  such  as  few
animals of  one  sex,  the lack  of  animal  survival  data,  animal  Infections and
0*780
                   VI11-35
                  07/31/91
-------
the  50% survival  rate among  the  high-dose group, combined with  the  lack  of

mlcropath>1og1c examination, must be noted In Interpreting these results.
                          HA -
                                                      (rounded to 10 mg/t)
where:
    125 mj/kg/day = NOAEL   In  male   rats   based  on  the  absence   of
                    Increased mortality and  hematologic  effects (Smith,
                    1953)

    10 kg         = assumed weight of a child

    100           = uncertainty  factor,  according  to'  U.S.   EPA   and
                    ODW/NAS  guidelines  for  use  with  a  NOAEL  from an
                    animal study

    1 l/day       = assumed water consumption by a child
              Longer-term HA . 125 mq/kq/daY  x  70  kq  =  ^
                 (adul              10D * * l/day      (rounded  to  40  mg/i)
where:
    125 rrg/kg/day = NOAEL   In   male   rats   based  on  the  absence   of
                    Increased   mortality   and   hematologlcal   effects
                    (Smith, 1953)

    70 kc         = assumed weight of an adult

    100           = uncertainty  factor,  according   to   U.S.   EPA  and
                    OOW/NAS  guidelines   for  use with a  NOAEL  from an
                    animal study

    2 l/cay       = assumed water consumption by an adult
    Assessment  of  Lonq-Term  Exposure  Data  and   Derivation   of  a  DUEL —

Smith (I'i53)  is also  the  only  available study for  the derivation of the DWEL

(U.S. EP/., 1991).  The DWEL is derived as follows:
                                     VIII-36
07/31/91
-------
RfD .
                        12'5
                                      .  0.125 rcg/kg/day
                                        (rounded to 0.1 mg/kg/day)
where:
    125 mg/kg/day

    1000
  NOAEL   In   male   rats   based  on  the  absence  of
  increased mortality and  hematologlc  effects  (Smith,
  1953)
  uncertainty  factor,  according  to  U.S.  EPA  and
  OOW/NAS  guidelines  for  use  with  a NOAEL  from  a
  subchronlc animal study
where:
    0.1 mg/kg/day
    70 kg
    2 l/day
                   nun     0.1  mq/kq/day  x 70  kg     _  ,.
                   DWEl  =          2 g/dav        *  3'5
                                    *   *           {rounded to 4 mg/i)
 = RfD
   = assumed weight of an adult
   = assumed water consumption by an adult
    Studies  Considered  for   Noncardnoqenlc   Quantification   —   PEP.    No
information was  available  on  the  effects  of  QEP  \n  humans.   Information  on
DEP toxidty in animals  1s  limited.   In  a  2-year  study (Food Research Labor-
atories, Inc.,  1955),  groups  of 30  rats  (15/sex)  were fed 0.5, 2.5  or  5.0%
levels  of  DEP  (250,  1250  or  2500 mg/kg  bw/day,  respectively)  In  the  diet.
No effects  were observed  at   levels   of  0.5  or 2.5%.   OEP at  the  5.0%  dose
level  resulted  In  a  small,  but significant decrease  In  the  growth  rate  of
the rats without any effect  on food  consumption.   Thus,  5.0% DEP appeared  to
affect  the  efficiency  of food  conversion  to  body mass.   No  Information was
available  on  the   numbers   of  rats  surviving  (42%  or   more  of  each  sex
survived)  the  2-year  study  period  and  hlstopathologlc  examination  was
performed  only  on  the  5.0%  dose  group.   Statistical   analysis  was  only
-------
conducted  on  organ weights and  excluded  statistically  higher rats from  the
respective  group  averages.  Also,  as  part of  this  study, 13 young  mongrel
dogs  were  fed DEP In the  diet at  levels  of  0, 0.5,  1.5,  2.0 and  2.5%  for  1
year.   Pr >blems were  encountered with  palatabllHy of DEP  1n  the  diet.  As  a
result,  tie  dogs  received varying  exposures  to DEP before  each dog attained
stablUza :1on  at   the  highest  tolerated  dietary  level.   Accordingly,  three
dogs  were  maintained at  0.5%,  one each  at  1.5 and  2.0%,  and  three at  the
2.5%  level.   The  average  weekly  Intakes of DEP were  computed  and  found  to be
0.8,  2.4,  3.5  and 4.4 g/kg/week  1n order  corresponding  to  Increasing  dietary
level.  N) effects were noted In dogs  as a result  of  DEP  exposures.

    Brown  et  al.   (1978)  also studied  the long-term  oral toxldty of DEP 1n
rats.   Groups  of   15  CD strain  rats  of each  sex were given diets  containing
0,  0.2,  1.0  or  5.0% DEP  for   16  weeks.  The authors estimated the mean
Intakes  to  be 0,   150,  770 and  3160 mg/kg/day  In  males  and 0, 150,  750  and
3710  mg/1-g/day In  females,  respectively.  Autopsies  and  hlstologk  exami-
nations  i'ere  conducted at the  end of 16 weeks.  No  changes 1n  behavioral
patterns or clinical  signs of toxldty were observed.   Female rats fed  diets
containing  1% DEP and  both  sexes  fed  diets  of 5% DEP  gained  significantly
less  welcht  than  the controls.   Mean  food consumption of  rats of  both  sexes
given  5%  DEP  and   females  given  1% DEP was significantly  lower than  that of
control rats.  In  order to Judge whether  palatabHUy was  the possible  cause
1n  decreased  weight  gain, a paired-feeding  study  was conducted.  Test rats
fed 5% DIP  consumed  more   food (total)  and gained  less  weight than controls.
Weights of  the brain, heart, spleen  and  kidney were significantly  lower 1n
male  and  female   rats  fed  5%  DEP.    Female   rats  given  5% DEP showed  a
statistically  significant Increase  In full  caecum weight.   There  were no
04780
V1II-38
07/02/91
-------
significant  changes  In  the absolute weights  of  any  organs  below the 5% DEP
dietary level.  Relative weights of the brain, liver, kidney,  stomach, small
Intestine and  full  caecum  were significantly higher  1n both sexes at the 5%
dietary level  when  compared with the  controls.   These  changes were attrib-
uted  to  the  compound-related  effect  on  growth  rate  since  dose-related
changes  In   gross  or  microscopic   pathology  were not  observed.   No other
effects were observed.

    Quantification of Noncardnoqenlc  Effects ~  DEP.
    Assessment  of  Acute  Exposure   Data  and  Derivation  of  the  1-Day HA  —
There were no satisfactory studies  for  the derivation of a 1-day  HA.

    Assessment  of  Acute Exposure  Data and  Derivation of  the 10-Day HA  —
There were no satisfactory studies  for  the derivation of a 10-day HA.
    Assessment  of  Longer-Term  HA —  There   Is  only  one  subchronlc  study
appropriate for  the  derivation of  longer-term HAs.  Brown  et  al.  (1978)  fed
CD rats diets containing  0.2,  1.0  and  5.0% DEP (150, 770 and 3160 mg/kg/day,
male;  150,  750  and 3710  mg/kg/day,  females)  for 16 weeks.  Female  rats  fed
1% DEP  and both  sexes  fed  5% DEP  gained significantly  less weight  than  the
controls.   A paired-feeding study  showed  this  weight difference was  not from
palatabllHy.  Both  the  liver  and  kidneys were  hlstologlcally  normal  at  all
DEP dietary  levels.   Relative kidney weights at the 5%  dose level were 0.67
and  0.69   g/100 g  bw  In  males  and  females,   respectively,   compared  with
control values  of  0.57  and 0.62  g/100 g bw  In males  and  females,  respec-
tively.   Although  slight  but  significant  (p<0.05)  changes   were  seen  In
females at the  1%  level,  the  use of   multiple  T  tests  for  the comparisons
04780
VIII-39
05/16/91
-------
(without  correction)  and  the small magnitude  of  the changes  Indicates  that
the  "\%  feeding  level  (750 mg/kg/day)  represents  a  NOAEL  1n  this  study.
Therefore,  the  NOAEL  (750 mg/kg/day,  females)  determined from the  Brown  et
al. (1978) study will be used to derive the longer-term HAs as follows:
                                750 mg/kg/day x 10 kg
               Longer-term HA
                  (child)
                    100 x 1  i/day
75 mg/S.
(rounded to 80 mg/a)
where:
    750 mc/kg/day


    10 kg
    1 i/d
-------
     Assessment  of  Lonf-terni  Exposure  Data  and  Derivation  of  a  DWEL —
 There  are two possible long-term studies for derivation  of  a lifetime DWEL.
 The  Brown et  al.  (1978)  16-week  study as  described for the longer-term HA 1s
 also considered  1n deriving  the  DWEL.   In a  2-year  dietary study,  Food
 Research  Laboratories,  Inc.  (1955) observed  similar  results at 5.0%  DEP as
 1n  the Brown  et  al.  (1978)  study.  They  reported  a NOEL  at 2.5% or  1250
mg/kg/day.  Deficiencies  'in  the  reporting of the  study  reduce confidence 1n
 the  use of  this data,  since  complete  Mstopathologles were not conducted and
no  Information was  available on   the  number  of  rats surviving  the  2-year
study by  Food  Research'Laboratories,  Inc.  (1955).   Therefore, the  NOAEL  (750
mg/kg/day,  females)  determined  from  the  Brown  et  al. (1978)  study will be
used to derive the lifetime DWEL (U.S. EPA, 1991).
    Step 1 - RfD Derivation
                  RfD
      750 mg/kg/day
           1000
0.75 mg/kg/day
(rounded to 0.8 mg/kg/day)
where:
    750 mg/kg/day = NOAEL  In  orally  exposed   rats  based  on  lack  of
                    kidney and weight gain effects (Brown et al., 1978)
    1000
= uncertainty  factor,  according  to  U.S.   EPA   and
  ODW/NAS  guidelines   for  use  with  a  NOAEL from  a
  subchronlc animal study
    Step 2 - DWEL Derivation
               m 0.8 mq/kq/day x 70 kq _                 '
                        2 I/day
where:
    0.8 mg/kg/day  = RfD
    70 kg          = assumed weight of an adult
    2 l/day        = assumed water consumption by an adult

04780
                   VIII-41
                             08/08/91
-------
    Studies  Considered   for   NoncardnogenVc   Quantification   —   PHP.    No
 Information was available on  the  effects  of  DMP 1n  humans.   The only  studies
 available on acute  oral  toxldty  1n  animals  used  lethality  as  the  toxU  end-
 point.   Tre  only  long-term  oral data was from  an unpublished  review  article
 (Lehman, 1955).

    Quantification  of Noncardnoqenlc  Effects  —  PHP.  The  1-day.   10-day
 or  longer-term HAs  and  a  lifetime  DWEL for  OHP cannot be derived  due  to
 Insufficient Information.

 Carcinogenic Effects
    There are  very few animal  carcinogenic  studies  on PAEs considering  the
 number  of  esters.   The  available human  studies  are  Inadequate due   to  the
 small  numters  of  subjects studied and  the  lack of  quantitative Information
 on  levels  and  duration   of  exposure.  The  human studies  were designed  to
 assess  todc  effects  caused  by  PAEs.   However,  there  Is  adequate  data  to
 consider !>EHP  to  be  a Group  82  compound (I.e.,  probable  human carcinogen)
 based  on significant  Increases 1n liver  tumor  responses  1n  rats and  mice of
 both  sexe;  .   B8P  has  been  classified as  a  Group C  compound (I.e., possible
 human  can  inogen)  based  on  mononuclear cell  leukemia  In female rats.   OBP,
 DEP  and  DMP  are  classified  as  Group  0   (I.e.,   not  classifiable)  since
 pertinent  data  regarding cardnogenlclty was  not  located   1n  the  available
 literature   (U.S.  EPA, 1986;    These  classifications have all  been  verified
 by the CR/VE Work  Group.

    Studlts Considered for  Carcinogenic  Quantification  —  DEHP.   In  an  NTP
 study  (19ii2a), 50 male and  50  female Fisher  344 rats per group were fed 6000
04780
VIII-42
08/08/91
-------
or  12,000  ppm  DEHP  In the diet for 103 weeks.  Similarly, groups of 50 male
and  50  female  B6C3F1  mice were given 3000 or  6000  ppm  OEHP  In the diet  for
103  weeks.  In  this  study  rodent  meal  was  provided  in  such a  way that
measured  food   consumption  actually  represented  significant  spillage   and
waste  rather  than   true  food   Intake.   For  this  reason  a  standard food
consumption  rate of  13%  of mouse and 5%  of  rat  body weight  was used  1n  the
dose conversion.   Corresponding dose levels  are  300 and  600  mg/kg/day  for
rats and  390 and 780  mg/kg/day  for  mice (low and  high dose,  respectively).
Doses were  those estimated  to be maximally and one-half maximally  tolerated
1n preliminary 90-day  Subchronlc feeding  studies.   The  animals  were -6 weeks
old  when  the study began, and  survivors  were sacrificed at  105 weeks.   All
animals were necropsled,  and a hlstologU  examination  of tissues  was made.
Treeted animals were compared with 50 matched controls of  each sex.

    Median  survival  times  were  >104  weeks for all  groups.   Body  weight  loss
was evident  In  low- or high-dose animals In each treatment group.

    A statistically  significant  {p<0.05 or  better)  Increase  1n the Incidence
of  hepatic  neoplasms  In  both  rats  and  mice  treated with  DEHP was  found as
described  In Table VIII-2.   Hepatic  tumors are described as hepatocellular
carcinomas  and  neoplastlc nodules 1n rats and  hepatocellular carcinomas  and
hepatocellular  adenomas  In mice.   Metastasis of liver  carcinoma  to the  lung
1n mice  was found   1n  5/50  high-dose males, 7/49  low-dose males,  7/50 high-
dose females and 2/50  low-dose females (NTP, 1982a).
    Carpenter  et  al.   (1953)  evaluated  the  chronic  toxlclty  of   OEHP  1n
Sherman  rats.   The  untreated  control  group and  each  treatment  group con-
04780
VIII-43
08/08/91
-------


























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04780
VIII-44
09/13/88
-------
 slsted  of  32  males  and  32  female  animals,  which  were  60 days  old  when  treat-
 ment  began.   Dosage  groups were given  400,  1300  or  4000 ppm DEHP  in  the diet
 for  1  year.   After  1 year of treatment, each group was  reduced to a maximum
 of  eight  males and  eight  females, and  treatment  was continued  for another
 year until sacrifice of the survivors.

    A  filial   (F.)  generation  of   animals  was produced  from  'animals  In  the
 control and  4000  ppm groups, which  had been given the  appropriate  diet  for
 120  days.   Each  Utter  was  reduced to  two males  and  two females  when  the
 pups  had  reached 15 days of  age, and  32  males  and  32  females In  the  F
 generation were  assigned to  each of  the  control  and 4000 ppm groups.   All
 surviving  F,  animals were sacrificed  after  being  maintained on control  or
 4000 ppm diets for 1 year.

    All animals were subjected  to necropsy  and  hlstopathologlc  examination.
 Only  40-47%  of the  animals   In  each  group,  Including   FI  animals,  survived
 1 year.   Of  the  animals  allowed   to  be on  study  for  2 years,  61-71%  died
 before  termination of the study at  2  years.   Lung Infection was diagnosed as
 the primary cause of death.

    No  malignant  tumors were observed In  this  study.   One  to five  rats  in
 each group (males and females combined}  had  tumors  with no treatment-related
 trend In evidence; however,  the  tumor  types were not  Identified.
    A carcinogenic  effect  of DEHP  was  not evident  1n  this  study.  However,
thH  study  Is weakened  by the  fact  that  of the  32 animals of  each  sex In
each  group  of   the  study  {excluding  the  FI  animals,  all  of  which  were
allowed to survive  1 year),  only eight  were allowed to survive beyond 1 year
04780
VIII-45
08/08/91
-------
 of  treatmj'nt.   Furthermore, mortality was  high with respect  to  all  groups.
 Hence,  an  Insufficient number  of  animals were  available  for  a  lifetime
 feeding  study of DEHP cardnogenlcHy 1n rats.

     Carperter et  al.  (1953) also  Investigated  the  toxldty  of DEHP 1n long-
 term studies In guinea pigs and  dogs.   Groups of  23  or 24  guinea  pigs  of
 each sex vere  fed 1300 or  4000 ppm  DEHP  1n the diet for  1  year until termi-
 nation  of the study.   Four dogs  were dosed 5  days/week  by  oral  administra-
 tion of  DEHP  In  capsules  at  a   dosage  of 0.03  ml/kg  for  the  Initial  19
 doses  followed  by  240  doses  at   0.06 ma/kg.   The  dogs  were  sacrificed  at
 the  end cf  the  1-year  dosing  period.   Pathologic  evaluation  of  the guinea
 pigs and dogs did  not  reveal  a  carcinogenic  effect of  DEHP.   However,  the
 treatment  and  survival  periods  for these  animals  were considerably  below
 their lifetimes.

     Since the animal  evidence  1s  considered by  U.S.  EPA to be sufficient and
 there 1s  10  human  data, according to the  U.S.  EPA Guidelines for Carcinogen
 Risk  Assessment,  OEHP  Is   classified as  a  B2  carcinogen (U.S.  EPA,  1986).
 This  classification  was verified  (10/07/87) by the CRAVE  Work  Group (U.S.
 EPA, 1991 .

     Quant flcatlon  of Carcinogenic  Effects —  DEHP.   The  risk  calculation
 1s  based on the  liver  tumor  data from  the   NTP  study  (1982a)  on  DEHP.
 Hepatocel lular carcinoma  and hepatocellular adenoma  Incidence were reported
 1n both  nale and female  rats  and male  and  female mice.  * However,  male mice
 were  the most  sensitive group.   The combined Incidence  of hepatocellular
 carcinoma; and  adenomas In male  mice  (see Table VIII-2) was 14/50 for con-
04780
VIII-46
08/08/91
-------
 trol  animals,  25/48 for  390  mg/kg/day animals  and  29/50 for 780  mg/kg/day
 animals.
    NTP  (19823),  Kluwe  et al.  (1982a), U.S.  EPA {19875}  and  IARC  (1982)
concluded  that  these   results  provide  sufficient evidence  of  d1(2-ethyl-
hexyl) phthalate-lnduced carclnogenlclty  In  rats  and  mice.   This  conclusion,
however,  1s  disputed.    Northrup  et al.  (1982)  claim  that  the NTP  (1982a)
results are  equivocal  since the MTD  was exceeded 1n some  treatment  groups,
Incidences of  liver tumors  varied within  different  control  groups  of  the
same  species  and   sex;  and  treated  animals   may have  been  malnourished.
Northrup et  al.  (1982) also claimed  that  the  rodent  data cannot  be  used  to
predict carcinogenic risk  In  humans because DEHP  1s  metabolized  differently
1n rats than In humans.  In response,  Kluwe  et al. (1983) noted that  the MTD
was not exceeded  since  there were  no  compound-related effects on survival,
the  Incidence  of   Hver   tumors   was   Increased  1n  OEHP-treated  animals
regardless  of   the   control  data  used  and   the   differences  In  metabolism
between rodents  and humans  would  not  affect  the carcinogenic response  In
rodents.  More  recently,  Turnbull  and  RodMcks  (1985) concluded  that  using
NTP (1982a)  data  to estimate OEHP-lnduced  carcinogenic risk  to  humans  will
probably  overestimate  actual   risk.   This  conclusion  was  based  on  the
differences  between rodents  and  primates  1n  the   metabolism  of  DEHP,  a
nonlinear  relationship  between  the  administered dose of  DEHP  to  the  dose of
the  "proximate  carcinogenic   species"   In   rodents,  the   fact   that  the
"proximate carcinogenic  species," which  1s  hypothesized to Induce cancer, Is
produced to  a  greater  extent  In rodents  than  In  primates and that there are
differences  In  target-site sensitivity between humans  and  rodents for liver
tumors  1n  general.
                                     VIII-47
08/08/91
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     The  dose-response data  used  1n  the potency calculations  Included  rats
with el .her hepatocellular  carcinomas  or  neoplastlc  nodules and  mice  with
either  lepatocellular carcinomas  or   adenomas  1n  the NTP  (1982a)  bloassay.
Hale ant!  female response data from the  rat and mouse were used to calculate
i
a  q-j*  falue  (Table  VIII-3).   The  oral  slope factors  were  3.18x10~'  and
4.52x10"'   (mg/kg/day)"1   for  male   and  female  rats,   and  1.41xlO~3  and
1.03x10"*   (nig/kg/day)"1   for   male  and   female   mice.    The   value   of
1.41x10"*  represents  the most  sensitive response and  hence 1s  selected  as
the  potency  value  for DEHP.   This  value  and the following risk/concentration
calculations   should  be  viewed   as   Interim   since  1t  would  appear  that
metabol sm  and pharmacoklnetU considerations  should  be  accounted for 1n the
dose  response  analysis.   The examination  of these factors  has  been  done  by
Turnbul I  and  RodMcks  (1985).  but has  not been  further evaluated  1n  this
documen;.   These  refinements will  be  further  evaluated  before  the  risk
values  are  put  to  final  use.  The upper-bound estimate of  the  cancer  risk
due  to  the  Ingestlon  of  2  1  of  water   for  a  70-year  lifetime  with  a
concent-atlon   of   contaminant   1s    4.0xlO~7   {wg/l)~l.    Since  risk   1s
assumed  to  be  linear  with  dose 1n  this range,  risk factors  of  10"*,  10"s
and  10~s correspond to 300, 30 and 3 vq/i, respectively.
    Studies  Considered  for Carcinogenic  Quantification —  BBP.   A bloassay
was performed  to  evaluate  the  carcinogenic  potential  of BBP 1n rats and mice
(Kluwe  et  al., 1982b;  NTP.  1982b).  Dietary  levels  of 6000  and  12,000 ppm
(780  and  1560  mg/kg/day)  BBP  were each  fed  to  groups  of  50  male  and  50
female F344  rats  and  50 male  and 50 female B6C3F1 nice.  Untreated groups of
50 mains and 50  females of each species were used as  controls.   The female
-------
-------
      TABLE VIII-3
Cancer Risk Calculations
Animal Dose
(mq/kg/day)
Animal/Se::
Rat/male
Rat/female
Mouse/mail?
Mouse/fem
-------
-------
 rats  and Doth  sexes  of  mice were maintained  on  these diets  for  103 yeeks;
 however,  ingestlon rates  and average  weights were  not  available  from the
 study.   The  male rats at both dose  levels  experienced high mortality within
 the  first  30  weeks  of  the  study,  at  which  time  the  male  rat  study was
 term1nate< .   No chronic  or  carcinogenic  effects  were observed  1n  male  or
 female  m1:e.  Among  female  rats,  however,  an Increase  In  mononuclear  cell
 leukemia uas observed at the higher dose level.

    Quant f1cation  of  Carcinogenic  Effects  —   B8P.    The  available  data
 meets  the criteria  for  limited  animal  evidence  based on  mononuclear  cell
 leukemia  In  female rats.   Hence BBP 1s  considered  to  be  a Group C, possible
 human  carcinogen  according  to  U.S.   EPA   Guidelines  for  Carcinogen  Risk
 Assessment   This  classification has  been  verified (08/26/87) by  the CRAVE
 Work GrouD  (U.S. EPA,  1991).   A  bloassay was performed by the NTP (1982b) to
 evaluate  the carclnogenUHy  of  BBP  In both rats  and mice.  The male rats at
 both dose  levels experienced  high mortality within the first 30 weeks of the
 study due  to apparent Internal  hemorrhaglng;  all  male rats were terminated
 at  30 weeks.   Among  female  rats  a  statistically  significant  Increase  In
 mononuclear  cell  leukemia  was observed at  the  high-dose  level  by comparison
 with  boti  concurrent controls  and  historical controls.   The  conclusions
 reached  Dy  the peer  review  group  of  this   study  Indicate   that  BBP was
 "probably"  carcinogenic  1n female  rats.   Although the Increase  In  leukemia
was stat  stlcally  significant,  the biological  relevance  of  this  finding was
questioned  due  to  the background  Incidence of mononuclear  cell  leukemia In
 Fischer  344 rats.  The  NTP  1s  currently  repeating  the  rat portion  of the
cancer bloassay for BBP.   Testing began In June, 1989 (NTP, 1991).
04780
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     Studies  Considered  forCarcinogenic  Quantification— DBF.   Pertinent
 data regarding  the  carclnogenlcHy  of  DBP could  not  be located  1n  the
 available  literature.  According  to U.S.  EPA guidelines  DBP 1s  classified  as
 Group  D,  not  classifiable.   This  classification  was verified  (08/26/87)  by
 the  CRAVE Work Group  (U.S. EPA, 1991).
    Studies  Considered for  Carcinogenic  Quantification  — D€P.   Pertinent
data  regarding  the  carclnogenlcHy  of  DEP  could  not  be located  1n  the
available literature.  According  to U.S.  EPA guidelines  DEP 1s  classified  as
Group  D,  not classifiable.   This classification was verified  (08/26/87)  by
the CRAVE Work Group  (U.S. EPA, 1991).

    Studies  Considered for  Carcinogenic  Quantification  -- PHP.   Pertinent
data  regarding  the  carclnogenlcHy  of  DMP  could  not  be located  1n  the
available literature.  According  to U.S.  EPA guidelines  DMP 1s  classified  as
Group  D,  not classifiable.   This classification was verified  (08/26/87)  by
the CRAVE Work Group  (U.S. EPA, 1991).
Existing Criteria and Standards
    The  American Conference  of  Industrial  Hyglenlsts  has  set a  TLV  of  5
     , as an 8-hour TWA, for  DEHP, DBP, DEP and DMP (ACGIH, 1985).
    The RfD Work  Group verified the following RfDs:  0.02  mg/kg/day for DEHP
(01/22/86);  0.2  mg/kg/day   for   BBP   (06/15/89);   0.1   mg/kg/day  for  DBP
(01/22/86); and 0.8 mg/kg/day  for  OEP  (07/16/87).   These assessments are all
available on IRIS (U.S.  EPA,  1991).   Quantitative  data are not available for
DMP.   The  CRAVE  Work  Group  has  verified  the  following  cancer  classlflca-
04780
VIII-51
08/08/91
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 Hons:  Group  B2 for  DEHP (10/07/87}; Group C for BBP (08/26/87); and Group D
 for  DBP,  D£P and  DMP  (08/26/87).   The oral slope  factor  for  DEHP,  the only
 one  of  thsse five phthalates  to have  a quantitative cancer risk Assessment,
 1s   4xlO~7   (yg/i)"1.    These  assessments  are   also   available   on  IRIS
 (U.S. EPA   1991).
 Interactionswith Other Chemicals
     PAEs  lave  been shown  to  Interact with other  compounds  In a synerglstlc
 or    antagonistic    manner.    Carbon    tetrachloMde,    barbiturates    and
 organophoiphate  Insecticides  (applied following PAE  exposure)  were shown to
 act  synerglstlcally with  PAEs  (Seth et  al.,  1979; Rubin and  Jaeger,  1973;
 Al-Badry  and Knowles,  I960}.   Antagonistic  effects were  noted  between PAEs
 (and  testlcular  zinc  levels),  methylenedloxyphenyl compounds,  paraoxon and
 simultaneously  applied  organophosphate   Insecticides  (Cater  et al..  1977;
 Foster  et  al.,  1980;  Melancon  and Lech, 1979;  Al-Badry  and Knowles, 1980).
 DEHP  has  been  shown  to  Increase  antlpyrlne metabolism  In rats, possibly by
 Inducing  hepatic  mlcrosomal  enzymes (Pollack  and Shen,  1984). Interaction
 between  i)EHP  and  ethanol  In rats has  been  studied  by   Agarwal  et  al.
 (1982a).   DEHP produces  changes 1n  the  pharmacologlc  response  to ethanol by
 altering ihe activities of alcohol dehydrogenase and aldehyde dehydrogenase.

    Agarwjl  et  al.  (1982b)  examined the  effects  of  DEHP  administration on
 phenobarbUal-lnduced  sleeping time 1n  rats.   The  authors  concluded   that
 PAEs  Interfere with blotransformatlon mechanisms of hepatic mlcrosomal drug-
 metabolizing  enzymes.   The  effects of  DEHP  on  the activity of  various
 enzymes differed between oral and  Intraperltoneal exposure routes.
04780
VIII-52
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 Special Groups at R1sk
     Patients  receiving  blood  transfusions  or  hemodlalysls   constitute   a
 high-risk  subpopulatlon  for  PAE  exposure.  This group rr.ay  receive  excessive
 quantities of PAEs  during  transfusion  or  hemodlalysls  due  to  leaching  of  PAE
 plastlclzers from plastic blood bags or plastic tubing.
    Hlllman  et  al.  (1975) studied the occurrence  of  necrotUIng  enterocoll-
t1s  and  OEHP  tissue  concentrations  1n  Infants who  had received  treatment
using arterial  catheters  containing  DEHP.   Higher  DEHP content was  found  In
catheterlzed Infants with necrotlzlng  enterocolHIs  than  In  Infants  that  had
been catheterIzed but did not  develop  this  disease.   While  the study did  not
show a causal  relationship,  It did demonstrate  that  DEHP accumulated In  the
tissues of critically 111 Infants.

    Gibson  et  al.  (1976) estimated  that the  amount  of DEHP  delivered to  a
patient during  hemodlalysls  ranged from 1.5-150 mg  for dialysis  lasting  15
minutes  to  5   hours.   Another  study  suggested  that  exposure  to  dlethyl
phthalate  during hemodlalysls  may  be  linked to  development of  hepatitis
(Neergaard et al.,  1971).   However,  evidence  of the  causal  relationship  was
not conclusive.
    It  1s  also possible that  workers  In the  manufacture  of PAEs  or  In the
plastics   Industry   constitute   a   high-risk   population.    However,  little
Information  1s available  for  these  groups.   The  only  prospective  cohort
sttdy  looked  at  workers  exposed   to  OEHP  for  periods  of  3  months  to  24
years.   This  study  did  not   demonstrate  any  compound-related   Injury  or
disease.   Therefore,  the  degree  of  risk  to  workers  cannot  be  quantified
(Thless et al., 1978b).
04780
VIII-53
08/08/91
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                                9.  REFERENCES

 Abe,  S.  ami M.  Sasaki.   1977.   Chromosome aberrations and  sister  chromatld
 exchanges  'n Chinese hamster cells  exposed  to various chemicals.   0.  Natl,
 Cancer Inst.  58: 1635-1641.

 ACGIH  (Ame'lcan  Conference  of  Governmental  Industrial Hyglenlsts).   1985.
 Documentation of  the  Threshold  UmH  Values  and Biological  Exposure Indices,
 2nd ed.  Cincinnati, OH.
                       /
 Agarwal, O.K.   1986.  Letters  to  the  Editor.   Toxlcol.  Appl.  Pharmacol.   82:
 383-385.
Agarwal,  O.K.,  S.  Agarwal  and  P.K.  Seth.   1982a.   Effect of  d1-(2-ethyl-
hexyl)  phthalate  on  drug  metabolism,  llpld peroxIdaUon,  and sulfhydryl  con-
tent of ral liver.  Drug Metab. Dlspos.  10(1): 77-80.
Agarwal,  O.K.,  S.  Agarwal  and  P.K.  Seth.   1982b.   Interaction of  d1-(2-
ethylhexyl   phthalate   with  the   pharmacological   response  and  metabolic
aspects of ethanol In mice.  Blochem. Pharmacol.  311(21): 3419-3423.

Agarwal, O.K., R.R. Haronpot,  J.C.  Lamb,  IV  and W.M.  Kluwe.   1985a.   Adverse
effects  o   butylbenzyl  phthalate  on  the   reproductive  and  hematopoletic
systems of male rats.  Toxicology.   35: 189-206.
Agarwal, O.K., W.H. Lawrence  and  0.  Autlan.   1985b.   AntlfertHKy and muta-
genlc  effects  In  mice  from  parenteral  administration  of  d1-2-ethylhexyl
phthalate [OEHP).  J.  Toxlcol. Environ. Health.  16: 71-84.
04790
IX-1
09/09/88
-------
Agar^al,   O.K.,   H.H.   Lawrence,   L.  0.   Nunez   and   J.   Autlan.   1985c.
MutagenUHy   evaluation   of   phthallc  add   esters   and  metabolites  In
Salmonella tjyphlmur jum cultures.   J.  Environ.  Health.   16:  61-69.

Agarwal,  O.K.,  S. Eustls,  J.C.  Lamb  IV,  J.R.  Reel and  M.H.  Kluwe.   1986.
Effects  of d1(2-ethylhexylJphthalate  on  the  gonadal  pathophyslology,  sperm
morphology  and  reproductive   performance  of  male  rats.   Environ.  Health
Perspect.  65: 343-350.

Al-Badry,  M.S.  and  C.0.  Knowles.   1980.  Phthalate-organophosphate  Inter-
actions: Toxlclty, penetration and metabolism studies  with  housefHes.  Arch.
Environ. Contam. Toxlcol.   9:  147-161.

Albro, P.W.   1986.  Absorption, metabolism  and  excretion  of d1(2-ethylhexyl)
phthalate by rats and  mice.  Environ. Health Perspect.   65:  293-298.

Albro,  P.W.   and  B.   Moore.   1974,    Identification  of the  metabolites of
simple phthalate dlesters  1n rat  urine.  J.  Chromatogr.  94: 209-218.

Albro, P.W.,  R.  Thomas and L. F1shbe1n.   1973.  Metabolism of  dlethylhexyl
phthalate  by  rats.  Isolation and  characterization of  the urinary  metabo-
lites.  J. Chromatogr.  76: 321-330.

Albro,  P.W.,  J.T.  Corbett,   J.L.  Schroeder,  S.  Jordan  and H.B.  Matthews.
198?.    Pharmacoklnetlcs,   Interactions   wHh  macromolecules   and   species
differences In metabolism of DEHP.  Environ. Health Perspect.  45: 19-25.
04790
IX-2
08/09/88
-------
Albro, P.M., J.T. Corbett,  J.L.  Schroeder  and  S.T.  Jordan.   1983a.  Incorpor-
ation  of  radioactivity From labeled d1-(2-ethylhexyl) phthalate  Into  DNA  of
rat liver  In vivo.  Chem.-B1ol. Interact.  44:  1-16.

Albro,  P.II.,  I.  Tondeur.  D.  Harbury,  S.  Jordan,  J.  Schroeder  and  J.T.
Corbett.   1983b.   Polar  metabolites  of d1-(2-ethylhexyl}  phthalate  In  the
rat.  Blociem. Blophys. Acta.  760: 283-292.

Aronson,  C.E.,  E.R.  Serllck  and G. Pretl.   1978.   Effects of  d1-(2-ethyl-
hexyl)  phthalate  on  the  Isolated  perfused  rat  heart.    Toxlcol.   Appl.
Pharmacol.  44: 155-169.  {Cited 1n Bell, 1982).

Ashby, J., F.J. deSerres, M.  Draper, et  al.   1985.   Overview and conclusions
of  the  IPCS  collaborative  study on Ui vitro  assay  systems.   lr\_:  Evaluation
of  Short-Term Tests  for  Carcinogens,  J.  Ashby, F.J.  deSerres,  H.  Draper,  et
al., Ed.  Elsevler Science Publishers,  Amsterdam,   p. 117-174.

Atlas,  E.,  R.   Foster and C.S.  Glam.    1982.   A1r-sea  exchange  of  high
molecular  weight  organic  pollutants:   Laboratory   studies.   Environ.  Scl.
Techno!.  16: 283-286.

Autian, J    1973.   Toxldty and health  threats of  phthalate  esters:  Review
of the literature.  Environ. Health Perspect.  3:  3-6.

Autlan,  J.   1982.   AntlfertUHy  effects  and dominant  lethal assays  for
mutagenk effects of DEHP.  Environ. Health Perspect.  45:  115-118.
04790
IX-3
08/09/88
-------
Bell,  P.P.   1982.   Effects  of  phthalate esters  on I1p1d metabolism in vari-
ous  tissues, cells  and organelles  In mammals.   Environ.  Health  Perspect.
45: 41-50.

Bell,  P.P.  and D.A.  Buthala.   1983.   Biochemical changes  1n  liver  of rats
fed  the plastldzer  d1-(2-ethylhexyl)  phthalate.   Bull.  EnvUonm.  Contam.
Toxlcol. 31: 177-182.

Bell,  F.P.  and  D.J.  Nazlr.   1976.   Effect of d1-(2-ethylhexyl) phthalate on
llpld  biosynthesis   In  selected  tissues  from rat  .In.  vUro.   Uplds.  11:
216-221. (Cited In Seth, 1982).

Bell,  P.P.,  C.S.  Patt,  8.   Brundage,  P.O.  Gillies and  W.G.  Phillips.  1977.
Studes  on  llpld biosynthesis  and cholesterol  content of  liver  and serum
llpcprotelns  1n rats  fed   various phthalate  esters.   Llplds.   13:   66-74.
(Cited In Seth, 1982).

BUckensdorfer, P. and L. Templeton.  1930.   A  study  of the  toxic  properties
of dlethylphthalate.  J. Am. Pharm. Assoc.   19:  1179-1181.

Bratt, H. and  P.  Batten.   1982.   Imperial  Chemical  Industries PLC.   (Cited
1n CPSC. 1985)

8ro*n. D.,  K.R. Butterworth, I.F.  Gaunt, P.  Grasso and  S.D.  Gangolll.  1978.
Short-term oral toxlclty study of  dlethylphthalate In the  rat.  Pood  Cosmet.
Toxlcol.  16: 415-422.
04790
IX-4
09/09/88
-------
Callahan,  M.A., M.W. SUmak, N.W. Gabel, et al.  1979.  Fate of 129 Priority


Pollutants  Vol.  II.   Office of  Water  Planning and  Standards,   U.S.  EPA,


Washlngtcn, DC.  (December),   p.  (94)1-28.





Calley,  ).,  0. Autlan and  W.  L. Guess.   1966.   Toxicology of a  series  of


phthalatt   esters.   J. Pharmacol.  Scl.   55:  158-162,   (Cited  In  GangolU,


1982; Seth, 1982),





Carpenter, C.P., C.S. Well and H.F. SmHh, Jr.   1953.  Chronic oral toxldty


of  d1-(2-ethylhexyl)  phthalate  for  rats,  guinea pigs  and  dogs.   AMA Arch.
                     t

Ind. Hyg.  Occup. Med.  8:  219-226.





Cater,  B.R.,  M.W.   Cook,  S.D.  GangolU  and  P.  Grasso.   1977.   Studies  on


dlbutyl  jhthalate-lnduced  testlcular  atrophy 1n  the  rat:  Effect on  zinc


metabolism.  Toxlcol. Appl.  Pharmacol.   41: 609-618.





Christian,  M.S.,   Ed.   1985.   Final   report  on  the  safety  assessment  of


dlbutyl phthalate,  dimethyl  phthalate and dlethyl phthalate.  J. Am. College


Toxlcol.  4(3): 267-303.





CMA  (Chemical  Manufacturing  Association).  1985.  A  21-day  feeding study  of


butyl  beizyl   phthalate  to   rats:  Effects on the liver  and  liver  llplds.


Report No. 0495/1/84.





CMA  (Chemical  Manufacturing  Asssodatlon.  1986.  A  21-day  feeding study  of


d1-N-butyl phthalate to rats: Effects on the  liver and liver I1p1ds.  Report


No. 0495/3/85.







04790                                IX-5                           09/09/88
-------
Cohen, A.J. and P.  Grasso.   1981.   Review  of the hepatic response to hydro-
Hpldaemlc drugs 1n rodents and assessment of Us toxicologlcal significance
to man.  Food Cosmet.  Toxlcol.   19:  585-605.

Corcoran,  E.F.   1973.   Gas-chromatographU  detection  of  phthaHc  add
esters.  Environ.  Health Perspect.   3:  13-15.

Coward, N.A.,  J.Mc.C.  Howell,  G.A.O.  PHt  and J.N.  Thompson.  1966.  Effect
of hormones on reproduction 1n  rats fed a  diet deficient  1n  retlnol  {vitamin
A alcohol)  but containing methyl  retlnoate  (vitamin A  add methyl  ester).
J. Reprod. Fertll.  12: 309-317.

CPSC  (Consumer  Products  Safety   Commission).   1983.   Children's   Chemical
Hazards   Risk   Assessment  on   D1(2-Ethylhexyl)   Phthalate  In  Children's
Products.  Chemical Hazards Program Directorate  for  Health Sciences.   August.

CPSC  (Consumer Products  Safety Commission).  1985.   U.S.   Consumer  Product
Safety Commission.  Report to  the  U.S. Consumer  Product  Safety  Commission  by
the Chronic Hazard Advisory Panel on d1(2-ethylhexyl)phthalate (DEHP).

Curto,  K.A.    1984.   Effects  of d1-(2-ethylhexyl)  phthalate and monoethyl-
hexyl phthalate on male  rodent  gonad.  Dissert.  Abst. Int. B.  45(1): 146.

Daniel,  J.W. and  H.  Bratt.   1974.  The absorption, metabolism and  tissue dis-
tribution  of  d1-(2-ethylhexyl)  phthalate  In   rats.   Toxlcol.   2:  51-65.
(Cited 1n  Seth, 1982).
 04790
IX-6
                                                                     09/09/88
-------
Darby,  J.  and J. Sears.   1969.   PlasUclzers.  IT>:  Encyclopedia  of Polymer
Science "echnology, Vol. 10, John Wiley and Sons, New York.  p. 228-306.

Dean,  Ed    1979.   Lange's  Handbook of Chemistry, 12th  ed.   McGraw-Hill Book
Co., New York.

Deangelo,  A.B.  and  C.T. Garrett.   1983.   Inhibition of development  of pre-
neoplastlc  lesions  1n  the  livers of rats  fed  a  weakly  carcinogenic  environ-
mental ccntamlnant.  Cancer Lett.  20: 199-205.

Deangelo,   A.B.,   C.T.  Garrett,   L.A.   Manolukas   and  T.   Yarlo.    1986.
D1-n-octyl  phthalate  {OOP},  a  relatively  Ineffective  peroxlsome  Inducing
straight  chain  Isomer  of  the  environmental  contaminant  d1(2-ethylhexyl)
phthalate  (DEHP), enhances  the development of  putative  preneoplastlc lesions
In rat liver.  Toxicology.   41:  279-288.

DeHaan, R.L.   1971.  Toxldty  of tissue  culture media  exposed  to  polyvlnyl-
chlorlde >last1c.  Nature.   231:  85-86.  (CHed In Bell, 1982)

Oming, d.L.   1977.   Interphase transfer processes.   II.  Evaporation  rates
of chlororoethanes,  ethanes, ethylenes, propanes  and propylenes from dilute
aqueous sjlutlons.  Comparisons  with  theoretical  predictions.   Environ.  Scl.
Technol.  11(4): 405-410.

DllUnghan,  £.0.  and  J.  Autlan.   1973.    Teratogenlclty,  mutagenldty  and
cellular  :oxU1ty of  phthalate  esters.  Environ. Health Perspect.   January.
p. 81-89.


04790                                IX-7                             09/13/88
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Oostal,  L.A.,  W.L.  Jenkins  and  8.A.  Schwetz.   1987a.   Hepatk  peroxlsome
proliferation  and hypoUpldemlc  effects  of  d1(2-ethylhexyl)  phthalate  In
neonatal and adult rats.   Toxlcol.  Appl.  Pharmacol.   87:81-90.

Oostal,  L.A.,   R.P.  Heaver   and  B.A.  Schwetz.    1987b.    Transfer   of
d1(2-ethylhexyl)phthalate through  rat  milk and  effects  on milk  composition
and mammary gland.  Toxlcol.  Appl.  Pharmacol.   91:  315-325.

Douglas, G.R.,  A.P.  Hugenholtz and  D.H.  Blakey.   1986.   Genetic  toxicology
of phthalate esters: Mutagenlc and other genotoxlc effects.   Environ.  Health
Perspect.  65:  255-262.

Dvoskln,  I.A.G.,   N.A.  Rokhmanlna,  F.V.  Demlyauko,  T.A.  Menshlkova,  L.F.
Ervreva  and  A.V.  Lashklna.  1969.   Hygienic  assessment  of certain  polymers
(provlnols).  G1g. Sanlt.  Jan:  34(1): 7-11.  (Translated  title)   (Ger.)

Elgenberg,   D.A.,  H.P.   Bozlglan,   D.E.  Carter  and   I.G.  S1pes.    1986.
Distribution, excretion  and  metabolism  of butylbenzl  phthalate In  the  rat.
J. Toxlcol. Environ.  Health.   17:  445-456.

E
-------
Engelhardt,  G.,  P.R.   Wallnoeter   and  0.  Hutzlnger.    1975.    Mlcroblal
metabolism  of  dlbutyl   phthalate  and  related  dlalkyl  phthalates.    Bull.
Environ. Contam. Toxkol.  13:  342-347.

Engelha-dt, 6., G.  Tlllmanns,  P.R.  Wallndfer  and 0. Hutzlnger.  1977.  Bio-
degradation of  d1-1sobutyl phthalate and related dlalkyl  phthalates by  Reni-
elli 1 urn niaclnum.  Chemosphere.   6:  347-354.

Ewlng,  J. and E.  Chlan.   1977.  Monitoring  to detect previously unrecognized
pollutants  In  surface  waters.   Office of Toxic  Substances, U.S.  EPA,  Wash-
ington, DC.  EPA 560/7-77/15a.

Food  Research  Laboratories,  Inc.    1955.   Data  submitted  to  U.S.  FDA  by
Celanese Corporation of America.   Report  No. 67567.

Foster,   P.M.,  L.V. Thomas, M.W.  Cook  and S.D.  Gangolll.  1980.   Study of
the  tesilcular  effects   and  changes  1n zinc  excretion  produced by  some
n-alkyl jhthalates 1n the rats.  ToxUol.  Appl.  Pharmacol.   54: 392-398.

Foster,  P.M.D.,  M.W.  Cook,  L.V.   Thomas,  et  al.   1982.   Differences  In
urinary   metabolic   profile   from   d1-n-butyl   pnthalate-treated   rats  and
hamsters   Drug Metab.  Dlspos.   11(1):  59-61.

GangolU,   S.O.    1982.   Testlcular   effects  of   phthalate  esters.   Environ.
Health P«rspect.  45: 77-84.
04790
IX-9
07/05/91
-------
Banning, A.E.,  U.  Brunk and  R.  Nllsson.  1981.   Induction of mitochondria
and peroxlsomes with phthalate esters.   Blochem.  Soc.  Trans.  9: 251P.

Gaining,  A.E.,  U.  Brunk  and  G.  Dallner.    1983.   Effects  of  dietary
d1-(2-ethylhexyl)  phthalate  on  the  structure and  function of  rat hepato-
cytes. Blochem. Blophys. Acta.  763:  72-82.

Gannlng, A.E.,  U.  Brunk and  G.  Dallner.   1984.   Phthalate esters and  their
effect on the IWer.  Hepatology.  4(3):  541-547.

Garvey,  L.K.,   J.A.  Swenberg,  T.E.   Hamm,   Jr.   and  J.A.  Popp.    1987,
01|2-ethylhexyl)phthalate:    Lack of  Initiating  activity   1n  the  liver  of
female F-344 rats.   Cardnogenesls.   8(2):  285-290.
G1amt C.S.,  H.S.  Chan,  G.S.  Neff  and E.L. Atlas.   1978.   Phthalate ester
plastldzers: A new class of marine pollutants.   Science.  199:  419-421.
Gibson, T.P..  W.A.  Brlggs and  B.J.  Boone.  1976.   Delivery of  d.1-2-ethyl-
hexyl phthalate to patients during hemodlalysls.  J. Lab. Cl1n. Med.   87(3):
519-524.

Gledhin,   W.E.,  R..G.  Kaley,  W.J.  Adams,  et  al.    1980.   An  environmental
safety  assessment  of  butyl  benzyl  phthalate.    Environ.  Sd. Techno!.   14:
301-305.
Glelberman,  S.E.,  I.A.  Kotova,  G.M.  Nlkolaev  and V.V.  Yurchenko.   1978.
Pharmacoklnetlcs of  dimethyl phthalate.   Med.  ParazHol.  Parazlt.  Boleznl.
47(3): 58-63.  (CA90: 80639m)
04790
IX-10
09/09/88
-------
Gollamudl,  R.,  H.R.  Prasanna,  R.H. Rao,  et  al.  1983.   Impaired  metabolism
of d1-(2-2thylhexyl)  phthalate (DEHP)  1n  old  rats—an  j£ vitro  study.   J.
Toxlcol. Lnvlron. Health.  12:  623-632.

Gollamudl, R.,  R.H.  Rao,  W.H.  Lawrence  and J.  Autlan.   1985.   Developmental
changes  1n  the  conversion rates of d1(2-ethylhexyl) phthalate  to  monoethyl-
hexyl phtialate 1n rats.  J. Toxlcol.  Environ. Health.   15:  459-465.

Graham,  I'.R.   1973.   Phthalate  ester  plastldzers  -  why  and  how they  are
used.  Environ.  Health Perspect.  3:  3-12.

Gray, T^.8,  n.d.   No title provided.   Unpublished work.  (Cited 1n  Foster
et al., 1982}

Gray, T.-J.B.  and J.A. Beamand.  1984.   Effect  of some phthalate  esters  and
other te; tlcular toxins on  primary cultures of  testlcular cells.   Food  Chem.
Toxic.  ;'2{2): 123-131.

Gray, T.3.B.  and S.O. GangolU.   1986.  Aspects  of  the  testlcular  toxlclty
of phthalate esters.  Environ.  Health  Perspect.   65:  229-235.

Gray, T.J.B.,  K.R.  Butterworth,  I.F.  Gaunt,  P.  Grasso  and  S.D. GangolU.
1977.   iihort-term  toxUHy  study of  d1-(2-ethylhexyl)  phthalate  In  rats.
Food Cosnet. Toxlcol.  15: 389-399.  (Cited In GangolU,  1982; Seth,  1982}.
Gray, T.J.B., I.R. Rowland, P.M.D. Foster and S.O.  GangolU.   1982.   Species
differences  In  the  testlcular  toxldty of phthalate esters.   Toxlcol.  Lett.
11: 141-147.
04790
IX-11
09/09/88
-------
Gray, T.J.B., 8.G. Lake, J.A. Beamand, et al.   1983.   Peroxlsomal effects of
phthalate esters  1n  primary cultures  of  rat hepatocytes.  Toxicology.  28:
167-179.
Gunn, S.A. and  T.C.  Gould.   1970.   Cadmium and other mineral elements,  in:
The Testls,  R.  0.  Johnson,  W. R. Gomes  and  N.  L.  Vandenmark, Eds., Vol. 3,
Academic Press, New York.   p.  377-481.   (Cited  1n Gangoin,  1982).

Harcano,  Y.,  A.  Kuwano,  K.   Inove  et  al.   1977.   Studies  on  toxlctty of
phthalk  acid   esters.    Part  I.    Teratogenlc    effects  In  mice   when
administered orally.   Osaka  Furltsu  Koshu  Elsel  Kenkyusho  Kenkyu hokoku,
Shokuhln Elsel  Hen.  8:  123-124.   (Abstract)

Harris,  R.S.,   H.C.   Hodge,   E.A.  Maynard  and  H.J.  Blanche!,   Jr.    1956.
Chronic  toxlclty  of  2-ethylhexyl  phthalate  In  rats  and  dogs.   Am.   Med.
Assoc. Arch.  Ind. Health.  13:  259-264.

Hattorl,  Y.,  Y. Kuge  and S.  Nakagawa.  1975.  M1crob1al  decomposition of
phthalate  esters  1n  environmental   water.    Pollut.   Control   Cent.   Osaka
Prefect.  M1zu Shorl  Gljutsu.   16:  951-954.

Hawley,  G.G.,  Ed.   1981.   The Condensed  Chemical  Dictionary, 10th ed.  Van
Nostrand Relnhold Co., New York.

Hlllman,  L.S.,  S.L.  Goodwin  and  W.R.  Sherman.   1975.   Identification and
measurement  of  plastldzer  1n neonatal tissue after umbilical catheters and
blood products.  New Engl. J.  Med.   292(8):  381-386.
04790
IX-12
09/09/88
-------
 HHes,  R.  1973.  Phthalates  In  the  Charles  and Merrlmack Rivers.   Environ.
 Health  'erspect.   3:  17-21.

 HHes,  R. and  K.  Bleman.   1972.   Water  pollution:  Organic  compounds In the
 Charles  River,  Boston.   Science.   178:  158.

 Hodgson   J.R.,  B.C.  Myhr, M.  McKeon  and  D.J.  Bruslck.   1982.  Evaluation of
 d1-(2-e
-------
Ishldate, H.,  Or.  and S,  OdasMma.   1977.   Chromosome  tests  with  134 com-
pounds on  Chinese  hamster cells  in  vMro -- A  screening  test for  chemical
carcinogens.  Mutat.  Res.   48:  337-354.

Ishlkawa, Y., K. Honda, S. Sasakawa, K. Hatada and H. Kobayashi.  1983. Pre-
vention  of  leakage  of d1-(2-ethylhexyl}  phthalate  from blood  bags  by glow
discharge  treatment  and  Its  effect  on  aggregablHty  of  stored platelets.
Vox Sang.  45: 68-76.

Jacobson, M.S., R.  Parkman,  L.N.  Button, R.J.  Yaeger  and  S.V. Kevy.  1974.
The toxlclty  of human serum  stored In flexible  polyvlnylchlorlde containers
on human flbroblast cell  cultures: An effect of  d1-(2-ethylhexyl) phthalate.
Res. Comm.  Chem. Pathol.  Pharmacol.   9(2): 315-323.

Jacobson, M.S., S.V.  Kevy and R.J. Grand.   1977.   Effects of a plastldzer
leached  from  polyvlnyl  chloride  on  the  subhuman primate:  A  consequence of
chronic  transfusion therapy.   J.  Lab.  CUn.  Med.  89:  1066-1079.  (Cited 1n
Seth,  1982).

Jaeger,  R.J.  and  R.J. Rubin.    1970.   Plastldzers  from  plastic  devices:
Extraction,  metabolism and  accumulation by  biological  systems.   Science.
170: 460-462.

Jaeger,  R.J.  and R.J.  Rubin.   1972.   Migration  of a phthalate  ester plastl-
c'zer   from  polyvlnyl  chloride  blood  bags  Into  stored human  blood  and  Us
location tn human  tissues. New Engl.  J.  Med.  287:  1114-1118.
04790
IX-14
07/05/91
-------
Jaeger, R.O.  and  R.J. Rubin.  1973.   D1-(2-ethylhexyl)  phthalate,  a plastl-
clzer contaminant of platelet concentrates.  Transfusion.  13(2): 107-108.

Johnson, E.N. and B.E.G.  Gabel.   1983.   An artificial  "embryo" for  detection
of abnormal developmental biology.  Fundam. Appl. Toxlcol.  3: 243-249.

Johnson, B.T.  and  W. Lulves.  1975.   Blodegradatlon of  d1-n-butyl  phthalate
and  d1-2-ethy1hexyl  phthalate 1n  freshwater  hydrosoll.   J.  Fish Res.  Board
Can.  3213): 333-340.

Johnson,  B.T.,  M.A. Heltkamp  and  J.R.  Jones.   1984.   Environmental  and
chemical factors  Influencing the  blodegradatlon of phthal1c-add  esters  In
freshwater sediments.  Environ.  Pollut. Ser. B. Chem.  Phys.   8(2): 101-118.

Jones,  A.E.,  R.H.  Kahn,   J.T. Groves  and E.A. Napier, Jr.   1975.   Phthalate
ester  toxlclty  In   human cell  cultures.   Toxlcol.   Appl.   Pharmacol.    31:
283-289.

Kaneshlma,  H.,  T.  Yamaguchl, T.  Oku1  and M. NaHoh.   1978.   Studies on  the
effects of  phthalate esters  on   the  biological  system  (Part  2}  -  hi  vitro
metabollsTi  and  biliary  excretion  of  phthalate  esters  In  rats.   Bull.
Environ. Contain. Toxlcol.  19: 502-509.

Katoh,  H  ,   S.  NakaJIma,  Y.  Kawashlma,  et  al.   1984.   Induction  of  rat
hepatic long-chain  acyl-CoA  hydrolases by  various  peroxlsome prollferators.
Blochem. 'harmacol.   33(7):  1081-1085.
04790
IX-15
07/05/91
-------
Kevy,  S.V.,  L.N.  Button  and M.S.  Jacobson.   1978.   Toxicology  of plastic
devices  having  contact  with blood.   Rep.  No. 1  HB  5-2906,  National Heart,
Lung and Blood Institute,  Bethesda,  Maryland.

Khan,  S.U.   1980.   Determining  the role of  humlc  substances  1n the fate of
pesticides  1n  the  environment.    International   Symposium   on  Hazards  of
Pesticides to the Environment and Human Health, Alexandria, Egypt,  Nov. 1-3,
1978.  J.  Environ.  Sd.  Health Pestle. Food  Contam.  Agrlc.  Wastes.  15(6):
1071-1090.

Khawaja, J. and  G.  Dallner.  1982.  Oral administration  of d1-{2-ethylhexyl}
phthalate  reduces  liver  protein synthesis  in  vivo.   IRCS Med. Sd.  10{8):
639.

Klhlstrom, I.    1983.   Placental  transfer of  dlethylhexyl  phthalate 1n  the
guinea pig placenta perfused ^n  sUu.   Acta Pharmacol.  ToxUol.   53:  23-27.

K1rby, P.E.,  R.F.  Plzzarello,   I.E.  Lawlor,  S.R.  Haworth  and J.R.  Hodgson.
1983.  Evaluation  of d1-{2-ethylhexyl) phthalate  and  Us major metabolites
1n  the Ames   test  and L51784  mouse  lymphoma  mutagenldty  assay.    Environ.
Hutagen.  5(5):  657-664.

Klausmeler,   R.E.   and  U.A.   Jones.   1960.   Mlcroblal   degradation  of
plastlclzers.  Develop.  Ind. Hlcroblal.   2: 47-53.
K'luwe,  W.M.   1982a.  Overview  of phthalate  ester  pharmacoklnetlcs  In mam-
malian species.  Environ.  Health Perspect.   45:  3-10.
04790
IX-16
07/05/91
-------
Kluwe, W,H.   19825.   Introduction.   Environ.  Health Perspect.  45: 1.

Kluwe,  fc.H.    1986.   Carcinogenic  potential of  phthaHc  add  esters  and
related   compounds:   Structure-activity  relationships.    Environ.   Health
Perspect   65: 271-278.

Kluwe, W M., O.K. Haseman, J.F. Douglas and  J.E. Huff.  1982a.  The carcino-
genldty  of  dietary  d1-(2-ethylhexyl)  phthalate (DEHP) 1n  Fischer  344 rats
and B6C3I, mice.  J.  Toxlcol.  Environ.  Health.   10(4-5): 797-815.

Kluwe,  h.H.,   E.E.  McConnell,  J.E.  Huff,  et   al.   1982b.   CardnogenlcHy
testing of phthalate esters and related compounds by the National Toxicology
Program  and  the National Cancer  Institute.  Environ.  Health Perspect.  45:
129-133.

Kluwe,  U.M.,   J.K.  Haseman  and  J.E.   Huff.  1983.   The  cardnogenlclty  of
d1-(2-etliylhexyl)  phthalate   (DEHP)  1n perspective.   J.   Toxlcol.  Environ.
Health.  12: 159-169.

Komarova   E.N.  1979.   Materials  on  the toxicology  of  dlbutyl  phthalate,
dloctyl  Dhthalate,  dlbutyl  sebacate and  butyl  stearate.   Tokaslkol.  SanH.
Khlm. Plrstmass.  3: 12-15 (translated).

Kom1towsl,1,  D., P.  Schmezer,  B.  Schmltt,   V.   Ehemann and  S.  Muto.   1986.
QuantltavWe  analysis  of  the early  changes  of  hepatocyte  nuclei  after
treating  Syrian golden  hamsters  with d1(2-ethylhexyl)  phthalate.   Cancer
Res.  Clln. Oncol.  Ill: 103-107.

04790                                IX-17                          07/05/91
-------
Kozumbo, W.J.,  R.  Kroll  and R.J.  Rubin.   1982.   Assessment of  the  mutagen-
UUy of phthalate esters.   Environ.  Health Perspect.   45:  103-109.

Krauskopf,  l.G.  1973.  Studies on the  toxlclty of  phthalates  via  Ingestlon.
Environ. Health Perspect.   3:  61-72.

Kurane,  R.,  T. Suzuki  and Y.  Takahara.    1979a.   M1crob1al   degradation  of
phthalate  esters.    IV.  Removal  of  phthalate esters  by  activated  sludge
Inoculated  with a  strain of Nocardla erythropolls.  Agrlc. B1ol.  Chem.   43:
421-427.

Kurane,  R.,  T. Suzuki  and Y.  Takahara.    1979b.   Mlcroblal   population  and
Identification  of  phthalate  ester-utilizing  microorganisms   In   activated
sludge Inoculated with microorganisms.  Agrl.  Blol. Chem.   43:  907-917.

Lake,  B.G.,  S.D.  Gangolll, P.  Grasso and  S.  Lloyd.   1975.   Studies on  the
hepatic  effects  of  orally  administered d1-(2-ethylhexyl)  phthalate In  the
rat.  Toxlcol. Appl. Pharmacol.  32:  355-367.   (Cited In Seth,  1982).

Lake,  B.G.,  P.G.  Brantum,  S.O.  Gangolll,  K.R.   Butterworth  and  P.  Grasso.
1976.  Studies  on  the  effects  of  orally admlstered d1-(2-ethylhexyl) phthal-
ate 1n the ferret.  Toxlcol.  6: 341-356.   (Cited  1n Seth.  1982).

Lake,  B.G.,  J.C.  Philips,  J.C.  Llnnell  and  S.D.  GangolH.   1977.  The .In.
vitro  hydrolysis of  some phthalate esters  by  hepatic  and  Intestinal  prepara-
tions  from various species.  Toxlcol. Appl. Pharmacol.  39: 239-248.
04790
IX-18
                                                                     07/05/91
-------
Lake, B.G.,  R.A.  Harris,  P.  Grasso  and  S.D. Gangollla.   1978.   Studies on
the  metabolism and  biological  effects of  n-butyl  benzyl  phthalate  1n  the
rat.   Prepared by  British  Industrial  Biological  Research Association  for
Monsanto, Report No. 232/78, June 1978.

Lake, B.G., T.J.B.  Gray,  J.R.  Foster,  et  al.   1984a.  Comparative studies on
d1-(2-etiylhexyl) phthalate-lnduced  hepatic peroxlsome  proliferation  In  the
rat and  lamster.  Toxlcol. Appl. Pharmacol.   72:  46-60.

Lake,  BG.,  W.R.  Pels  Rljcken,  T.J.B.  Gray,  et  al.  1984b.   Comparative
studies  of  the hepatU  effects of  d1- and mono-n-octyl phthalates,  d1-(2-
ethylhex/1) phthalate  and chloflbrate 1n  the rat.   Acta Pharmacol.  Toxlcol.
54: 167-176.

Lake, B.3., T.  Gray and S.O. GangolH.   1986.  Hepatic  effects  of phthalate
esters aid related  compounds  - _1_n vivo and j_n vitro correlations.  Environ.
Health P'rspect.  67: 283-290.

Lamb, J.C.,  IV, R.E.  Chapln,  J.  Teague,  A.O. Lawton and  J.R.  Reel.   1987.
Reproducilve effects  of  four  phthallc  ac'ld esters  1n  the mouse.   Toxlcol.
Appl. Ph-irmacol.  88(2}: 255-269.

Lawrence  W.H.  and  S.F.  Tuell.   1979.   Phthalate  esters:  The   question  of
safety - an update.   Clln. Toxlcol.  15(4):  447-466.
04790
IX-19
07/05/91
-------
Lawrence, W.H.,  M.  Malik, L.C.  Turner,  A.R.  Singh and J.  Autlan.   1975.   A
tox1co!og1cal  Investigation of  some  acute,  short-term  and  chronic  effects  of
administering  d1-(2-ethy1hexyl)  phthalate  (DEHP)  and other phthalate esters.
Environ. Res.  9: 1-11.

Leah,  T.O.   1977.   Environmental Contaminants  Inventory  Study  No, 4:  The
Production, Use  and  Distribution of  Phthallc  Add Esters  1n  Canada.  Report
Ser. No. 47.   Inland Haters  Directorate, Ontario Region,  Water  Planning  and
Management Branch, Burlington, Ontario.

Lefaux,  R.   1968.   Practical  Toxicology  of  Plastics,  CRC  Press,  Chemical
Rubber Co., Cleveland, Ohio.   (Cited  1n Krauskopf, 1973).

Lehman,  A.J.   1955.   Insect  repellents.   Food  Drug  Office  Q.  Bull.   19:
87-99.

Lehman,  A.J.   1959.   Appraisal 'of  the  safety  of chemicals  In  foods,  drugs
and cosmetics.   Assoc. Food Drug Officials  United States

Lewandowskl, M., J.  Fernandes  and  T.S.  Chen.   1980.    Assessment  of  the
teratogenlc  potential  of  plasma-soluble extracts of  dlethylhexyl  phthalate
plastldzed polyvlnyl  chloride  plastics  In rats.  Toxlcol.  Appl.  Pharmacol.
54: 141-147.
Lhuguenot,  J.C.,  A.M.  Mitchell,  G.  MUner,  E.A.  Lock  and  C.R.  Elcombe.
1985.    The   metabolism    of    d1(2-ethylhexyl)    phthalate    {DEHP}    and
mono(2-ethylhexyl) phthalate  (MEHP)  1n  rats: In.  vivo  and In vitro  dose  and
time dependency of metabolism.  Toxlcol.  Appl.  Pharmacol.   80:  11-22.
04790
IX-20
07/05/91
-------
Llndgren, A.,  N.G.  L1ndqu1st,  A.  Lyden,  et al.   1982.   A whole body  auto-
radlographU  study  on  the  distribution  of  14C-labelled  d1-(2-ethylhexyl)
phthalate In mice.  Toxicology.   23:  149-158.

Lyman, W  J.,  H.F. Reehl and  O.H.  Rosenblatt.   1982.   Handbook of  Chemical
Property Estimation  Methods.   McGraw-Hill  Book  Co.,  New York.   p.  4-9,  15-16.

Mabey, W.R.,  3.H. Smith,  R.T.  Podoll,  et  al.   1982.   Aquatic fate process
data  for  organic priority pollutants.   Prepared by  SRI  International  for
U.S.  EPA,  Office of  Water  Regulations  and Standards,  Washington,  DC.   EPA
440/4-81-014.

Mallette   F.S.  and  E.  Von Haam.  1952.   Studies on  the  toxIcHy  and  skin
effects of compounds  used  In  the rubber and plastics  Industries.  I. Plastl-
dzers. Arch. Ind. Hyg.  Occup. Med.  6:  231-236.

Mangham,  8.A., J.R. Foster and  B.G.  Lake.  1981.  Comparison  of  the hepatic
and  tesilcular  effects  of orally  administered  d1-(2-ethylhexyl)  phthalate
and dlalcyl 79 phthalate In the  rat.   Toxlcol.  Appl.  Pharmacol.   61:  205-214.

Marcel, Y.L.   1973.   Determination  of  d1-(2-ethylhexyl)  phthalate   In  human
blood  plasma  and cryopreclpltates.   Environ.  Health  Perspect.  3:  119-121.
(Cited IT Pollack et al., 1985b)

Marcel, Y.L. and  S.P. Noel.   1970.   A plastlclzer  In  Upld  extracts  of  human
blood.  Chem. Phys.  Llplds.  4:  418-419.
-------
Hark,  H.F.,  D.F. Othmer,  C.G.  Overberger,  G.T.  Seaborg,  Ed.   1982.  Klrk-
Othmer Encyclopedia of  Chemical  Technology,  3rd ed.( Vol. 17.  Wiley  Inter-
science Publication, John Wiley and Sons.   p.  745.
Mason, K.E.   1933.   Differences  In  testes  Injury and repair after  vitamin  A
deficiency, vitamin E deficiency and Inanition.   Am.  J.  Anat.   52:  153-239.

Mathur,  S.P.   1974.   Phthalate  esters  In  the  environment:  Pollutants or
natural products?  J. Environ. Qual.  3:  189-197.

Matsuda,  K. and  H.  Schnltzer.  1971.   Reactions between fulvlc add, a  soil
humlc material and  dlalkyl  phthalates.   Bull. Environ. Contam. Toxlcol.  6:
200-204.

Mayer, F.L.   1976.   Residue dynamics  of d1-(2-ethylhexyl) phthalate  In  fat-
head minnows (Plmephales promelas).   3.  Fish.  Res.  Board Can.   33:  2610-2613.

Melancon, M.J.,  Jr.  and J.J. Lech.   1979.   Structural  requirements  for  the
Inhibition of phthalate ester hydrolysis 1n rainbow  trout  by methylenedloxy-
phenyl compounds.  Xenoblotlca.   9:  317-322.

Melnlck,  R.L. and C.M.  Sctilller.  1982.  MHochondrlal  toxldty of  phthalate
esters.  Environ. Health Perspect.  45:  51-56.
Nen'shlkova, T.A.   1971.   Hygienic  evaluation of dlbutyl phthalate  In  rela-
tion  to  the  use of  polymeric materials  for finishing  living quarters  on
ships.  Gig. Sanlt.   36: 23-27  (Translation)
-------
Mllkov,  I.E.,  M.V. Aldyreva,  T.B.  Popova,  et  al.   1973.  Health  status  of
workers  exposed  to phthalate  plastldzers  In the manufacture  of  artificial
leather  and  films based  on  PVC  resins.    Environ.  Health  Perspect.   3:
175-178.

Mitchell, F.E.,  S.C.  Price,  R.H.  Hlnton, P.  Grasso and  J.W.  Bridges.   1985.
Time  anl dose-response  study  of  the  effects  on  rats  of  the  plastldzer
d1(2-ethylhexyl) phthalate.  Toxlcol. Appl. Pharmacol.  81: 371-392.

Monsanto.  1972.  Unpublished work.   (Cited 1n Krauskopf, 1973)

Moorhead, P.S.,  P.C. Nowell, W.J. Mellman,  D.M.  Battlps  and  D.A.  Hungerford.
1960.   Chromosome  preparations of  leukocytes cultured from  human  peripheral
blood.   Exp. Cell Res.  20(3):  613-616.   (Cited 1n Thless and Flelg,  1979)

Murakami, K. and K. NIshlyama.  1986.  Toxlclty  of  dlbutyl phthalate and  Us
metabolites 1n rats.  Jap.  J. Hyg.  41(4):  775-780.

Mushtaq, M.  and  K.K.  Oatta.   1981.   Effect of dl-2-ethylhexyl phthalate  on
rat testls.   Indian J. Blochem. Blophys.   18(4.Suppl.}:  159.   (Abstract)

Nalr,  N.  and  C.K.R.  Kurup.   1986.    A  comparative  study of  the  effects  of
admlnlst-atlon of  dlethylhexyl  phthalate on hepatic mitochondria  of  the  rat
and the mouse.  Indian J.  Blochem. Blophys.   23:  270-273.

NAS  (Na .1onal  Academy  of  Science).    1977.   Drinking  Water and  Health.
National Academy Press,  Washington,  DC.   Vol. I.   p.  19-63.


04790                                IX-23                           07/05/91
-------
NAS  (National  Academy  of  Science).    1980.    Drinking  Water  and  Health.
National Academy Press, Washington, DC.  Vol. 3.  p. 25-67.

Nazlr,  D.,  A.P.  Alcaraz,  B.A.  Blerl,  M.  Beroza  and P.P. Nalr.  1971.  Isola-
tion,  Identification  and specific localization  of  d1-(2-ethylhexyl) phthal-
ate  In  bovine heart muscle mitochondria.  Biochemistry.  10: 4425-4429.

Neergaard,  J.,   B.  Nielsen,  V.  Faurby,  D.H. Chrlstensen  and  O.F.  Nielsen.
1971.   Plastlclzers  In PVC and the occurence of hepatitis  In a hemodlalysis
unit.   Scand. J. Urol. Nephrol.  5: 141-145.

Nlkonorow,   H.,   H.   Mazur  and   H.   Plekacz.    1973.   Effect   of  orally
administered  plastlclzers and polyvlnyl  chloride  stabilizers  In  the  rat.
To;<1col. Appl. Pharmacol. 26:  253-259.   (Cited  In Seth, 1982)

NIOSH  (National  Institute for  Occupational  Safety and Health).  1985.  RTECS
(Registry of  Toxic  Effects of  Chemical Substances) master  file listing a? of
Harch,  1984.   U.S.  Department of Health  and  Human  Services,  Public Health
Services, Center of Disease Control, NIOSH,  Cincinnati, OH.

.Northup,  S.,  L.  Martls, R. Ulbrlcht,  et al.  1982.  Comment on the carcino-
genic   potential  of  d1-(2-ethylhexyl)  phthalate.   J.  Toxlcol.  Environ.
Health.   10:  493-518.
 NTP   (National  Toxicology   Program).   1982a.   Carclnoglnesls  Bloassay  of
 d1(2-ethylhexyl)phthalate  (CAS No.  117-81-7)  In  F344 Rats  and  B6C3F]  Nice
 (Feed  Study).  NTP Tech. Rep.  NIN/PUB-82-1773,  NTP-80-37.  NTIS P882-184011.
 04790
IX-24
07/05/91
-------
NTP  (National  Toxicology  Program).   1982b.   Carclnogenesls  Bloassay  of
Butylbenz^lphthalate  (CAS  No.  85-68-7)  In  F344  Rats  and B6C3F   Mice  (Feed
Study). NIP Tech, Rep. NTIS PB83-118398, Research Triangle Park, NC.  98 p.

NTP  (National  Toxicology  Program).   1984a.    D1-(2-ethylhexyl)  phthalate:
Reproduct on  and  fertility assessment  1n  CO-1 mice  when  administered  by
gavage.  I 1nal  Report.  NTP, Research Triangle Park, NC.

NTP    (National   Toxicology    Program).     1984b.    01(n-Butyl)Phthalate:
Reproduction  and fertility assessment  In CO-1 mice when  administered in  the
 feed.   F1ial  Report.   NTP,  Research  Triangle  Park,  NC.

 NTP  (National  Toxicology  Program).   1984c.   Dlethylphthalate:  Reproduction
 and  fertility  assessment  In CO-1  mice  when administered In  the  feed.   Final
 Report.  fTP, Research Triangle Park,  NC.

 NTP (National  Toxicology Program).   1985.  Twenty-six week  subchronlc  study
 and  modified  mating  trial  In  F344  rats.   Butylbenzylphthalate.   Final
 report.   Project  No.  12307-02,  -03.   Hazelton  Laboratories America,  Inc.
 Unpublished  report.

 NTP  (Nat  onal  Toxicology  Program).   1986.  Addendum to  final  report.   Bone
 marrow differential  results  —   26-week   study.    IBI/HLA  Project  No.
 12307-02.  Hazelton  Laboratories  America.  Inc.   Unpublished  report.
  NTP
(National  Toxicology Program).   1991.   Chemical  Status  Report.   07/09/91.
  04790
                                       Ix_25                           08/08/91
-------
Qgner,  6.  and M.  Schnltzer.  1970.   Humlc  substances: Fulvk  add-d1a1kyl
phthalate complexes and their role 1n pollution.   Science.   170:  317-318.

Q'Grady,  D.P.,  P.M.  Howard  and  A.F.   Werner.    1985.    Activated  sludge
blodegradatlon   of   12   commercial   phthalate   esters.    Appl.   Environ.
H1crob1ol.  49(2): 443-445.

Qhta,  Y.  and  M.   Nakamoto.   1979.   Metabolism  of  d1-n-butyl  phthalate  by
Aeromonas sp.  Hakkokogaku.  57:  50-53.

Qlshl, S.  1984a.  Effects of d1-2-ethylhexyl  phthalate on  llpld  composition
of serum and testls In rats.   Toxlcol. Lett.   23:  67-72.
01sh1, S.   1984b.   Testlcular atrophy  of  rats Induced  by  d1-(2-ethylhexyl)
phthalate: Effects of vitamin A and zinc concentrations  In  the  testls,  liver
and serum.  Toxlcol.  Lett.  20:  75-78.
01sh1,  S.   1986.  Testlcular  atrophy  Induced  by  d1(2-ethylhexyl)phthalate:
Changes   In   histology,    cell    specific   enzyme   activities   and   zinc
concentrations 1n rat testes.  Arch. Toxlcol.  59:290-295.

O'ishl,  S.  and  K.  Hlraga.   1982.   Distribution  and  elimination  of  d1-(2-
ethylhexyl) phthalate  (OEHP)  and mono-2-ethylhexyl phthalate  (MEHP)  after  a
single oral administration of OEHP 1n rats.   Arch..  Toxlcol.   51:  149-155.
Qishl, S.  and  K. Hlraga.   1983.   Effects  of d1-(2-ethylhexyl)  phthalate  on
llpld composition  of liver,  testls  and serum of  male  rats.  Jpn  J.  Pharm.
33(Suppl.): 153.  (Abstract)
04790
IX-26
08/08/91
-------
OkHa,  R.  and  C.  Chance.   1984.   Induction  of   laurate  w-hydroxylase  by
d1-(2-etfcy1hexyl) phthalate In rat  liver mlcrosomes.   Blochem.  Blophys.  Res.
Comm.  121(1): 304-309.
Osuml, T  and  T.  Hashimoto.   1978.   Enhancement of fatty acyl-CoA  oxidizing
activity  In  rat  peroxlsomes  by  d1-(2-ethy1hexyl) phthalate.   J.   Blochem.
83: 1361-1365.   (CHed In Seth, 1982).

Parmar,  )., S.P.  Srlvastava,  S.P.  SMvastava  and P.K. Seth.  1985.   Hepatic
mixed function oxldases and cytochrome P-450 contents  In rat  pups exposed  to
d1-(2-etliylhexyl}phthalate  through   mother's   milk.    Drug.   Metab.   Olspos.
13(3): 31)8-370.
Parmar,   D.,    S.P.    Srlvastowa   and   P.K.   Seth.    1986.     Effect   of
dl(2~ethflhexyl)phthalate   (DEHP)    on   spermatogenesls   In   adult  rats.
ToxUolojy.  42: 47-55.
Peakall, 0.8.   1975.   Phthalate esters:  Occurrence  and biological  effects.
Residue ?ev.  54:  1-41.

Peck,  C.C.   and  P.W.  Albro.   1982.    Toxic  potential  of  the  plastlclzer
d1-(2-ethylhexyl)  phthalate  In the* context of Its disposition and  metabolism
In primates  and man.   Environ.  Health  Perspect.   45:  11-17.

Peck,  C C.,  O.G.  Odom,  H.I. Friedman.   1979.   D1-(2-ethylhexy1)  phthalate
(DEHP)  and  mono-2-ethylhexyl phthalate  (MEHP)  accumulation  In whole  blood
and red cell concentrates.   Transfusion.  19:  137-146.
                                                                       /nc /
-------
Perez,  3.A.,  M.A.  Herriandex,  R.A.  Ruiz  and   P.J.   Brown.    1977.   The
utilization of  the plastlclzer dimethyl  phthalate  by  an Isolated strain  of
Enterobacter aeroqenes.  Bull. Environ.  Contain.  Toxlcol.   18:  104-107.

Peters, J.W. and R.M.  Cook.   1973.   Effects  of  phthalate  esters  on reproduc-
tion 1n rats.  Environ. Health Perspect.   3:  91-94.

Petersen,  R.V., D. Lyman, D.B. Roll  and  E. Swlnyard.   1972-1975.  Toxicology
of plastic devices having contact with blood.   Final report.   NIH, Bethesda,
HO.  NIH-NHLI73-2908-B.  (Cited In  Bell,  1982)
Phillips, B.J., T.E.B. James and S.D.  Gangolll.   1982.   Genotoxldty  studies
of  d1-(2-ethylhexyl)  phthalate and  Us  metabolUles  In  CHO cells.   Mutat.
Res. 102: 297-304.

Plllal, K.S.R.  and  P.K.  Seth.  1978.   Influence  of low protein diet  on  the
toxlclty  of   d1-{2-ethylhexyl)  phthalate.    Ind.   J.  Blochem.  Blophys.
16{Suppl.) Abstr.  No.  243.   (CHed  1n Seth,  1982)
Plasterer, M.R.,  H.S.  Bradshaw,  G.M.  Booth,  M.H.  Carter,  R.L.  Schuler  and
B.O. Hardln.   1985.   Developmental  toxlclty of nine selected compounds  fol-
lowing  prenatal  exposure  1n  the mouse:  Naphthalene,  0-n1trophenol,  sodium
selenlte, dimethyl phthalate, ethylenethlourea and  four  glycol ether  deriva-
tives.  J. Toxlcol. Environ. Health.   15:  25-38.
Pollack,  G.M.  and  D.D.  Shen.   1984.   Effect  of renal  failure and  b1s(2-
ethylhexyl) phthalate pretreatment on the dlspostlon and  metabolism  of antl-
pyrlne In the rat.  0. Pharm. Scl.  73(1):  29-33.
                                     TV-9R
07/05/91
-------
Pollack, G.H., R.C. LI,  J.C.  Ermer  and D.D. Shen.  1985a.  Effects  of  route
of administration and repetitive dosing on the disposition kinetics  of  d1(2-
ethylhexyl) phthalate and  Its  mono-de-esteMf1ed metabolite In rats.   Tox1-
col. Appl. Pharmacol.   79:  246-256.

Pollack,  G.M.,  J.F.  Buchanan,  R.L.  Slaughter,  R.K.  Kohlll,   D.D.   Shen.
1985b.   Circulating  concentrations   of  d1-(2-ethylhexy1) phthalate  and  Us
de-ester Ifled  phthallc   add   products   following   plastldzer  exposure   1n
patients receiving hemodlalysls.  Toxlcol.  Appl.  Pharmacol.  79: 257-267.

Putman,   D.L.,   W.A.  Moore,   L.M.   Schechtman  and  J.R.  Hodgson.    1983.
Cytogene;1c evaluation of  d1-2-ethyl  phthalate and Us major metabolites  1n
Fischer :i44 rats.  Environ. Mutagen.   5(2):  227-232.

Reddy, 3 K., D.E. Mody,   D.L. Azarnoff and M.S. Rao.   1976.  01(2-ethylhexy1)
phthalatt :  An  Industrial   plastldzer  Induces  hypollpldemla  and   enhances
hepatic  catalase  and  carn1t1n-e  acetyltransferase activities  In  rats  and
mice.  Life Sc1.   18:  941-945.   (Cited In  Seth, 1982).

Reddy,  J.K.,   M.K.  Reddy,  M.I.  Usman.  N.O.  Lalwanl.  and  S.  Ras.   1986.
Comparlscn  of  hepatic peroxlsome  prollferatlve  effect  and  Us   Implication
for  hepatocardnogenldty  of phthalate esters, d1(2-ethylhexyl)phthalate  and
d1(2-ethylhexyl)   adlpate   with  a   hypollpldemlc   drug.   Environ.  Health
Perspect.  65: 317-327.
04790                                IX-29                           07/05/91
-------
Rhodes,  C.,  T.C. Orton,  I.S.  Pratt,  et  al.   1986.   Comparative  pharmaco-
klnetlcs and  subacute  toxkUy  of d1(2-ethylhexyl) phthalate (DEHP) in  rats
and marmosets:   Extrapolation of  effects  1n  rodents to man. Environ. Health
Per spect.  65: 299-308.
Rodrlcks,  J.V.   and   0.   Turnbull.   1987.   Interspecles  differences   In
peroxlsomes and  peroxlsome  proliferation.   Toxlcol. and Ind. Health.   3(1):
197-212.

Rowland,  I.R.    1974.   Metabolism  of  d1-(2-ethylhexyl)  phthalate  by  the
contents of  the  alimentary  tract  of  the  rat.   Food  Cosmet.  Toxlcol.   12:
293-302.

Rowland,  I.R.,  R.C.   Cottrell  and J.C.  Phillips.   1977.   Hydrolysis   of
phthalate esters  by the  gastrointestinal  contents of the rat.   Food  Cosmet.
Toxlcol.  15:  17-21.

Rubin,  R.J.    1975.    Metabolism  and  acute  lung  tox1c1ty  of   solubllized
d1-(2-ethylhexyl) phthalate  (DEHP)  In  rats.   Irr. Mechanisms of  Tox1c1ty  and
Metabolism.   Proc.  Sixth  Int.  Congr.  Pharmacol.   Helsinki,  Finland.   6:
205-213.

Rubin,  R.J.  1976.  Transcript  of proceedings.   Workshop on adenlne  and  red
cell preservation.  Food Drug Admin. Bureau B1ol.. DHEU.
04790
IX-30
07/05/91
-------
Rubin, R 0. and  Chang.   1978.  Effect of  Intravenous  administration of the
solublzlllzed   plastldzer,   d1(2-ethylhexyl)phthalate,   on   the  lung  and
survival   of  transfused  rats.   Presented   at  17th  Ann.  Meet.   Society  of
Toxicology.  (Abstract)

Rubin, F.J. and  R.J.  Jaeger.   1973.   Some pharmacologlc  and   toxlcologlc
effects  of  d1-(2-ethylhexyl)  phthalate   (OEHP)  and  other  plastldzers.
Environ.  Health Perspect.   3:  53-59.   {CHed 1n Bell, 1982).

Rubin, R.J. and P.P. Na1r.  1973.  Plastldzers In human  tissues.  New Engl.
J. Med.  288:  915-916.

Rubin,  *.   and  C.  Schlffer.    1976.   Fate  In  humans  of  the  plastlclzer,
d1-(2-ethylhexyl) phthalate, arising from transfusion of platelets stored In
vinyl plastic  bags.  Transfusion.   16:  330-335.

Rubin, i.J.,  H.  Kozumbo  and  R.  Kroll.   1979.   Ames  mutagenlc  assay  or  a
series of   phthallc  add  esters:   Positive response  of  the dimethyl  and
dlethyl  esters  In  TA 100.   Soc.  Toxlcol.  Ann.  Meet.,  New  Orleans,  March
11-15.  p.  11. (Abstract)

Ruddlck  J.A., O.C. Vllleneure,  I.  Chu,  E.  Nestman and 0. Miles.  19.81.  An
assessment  of  the  teratogenldty  In the rat and mutagenldty 1n Salmonella
of mono-2-ethylhexyl  phthalate.   Bull.  Environ.  Contam.  Toxlcol.   27:  181.
(CHed in Thomas  and Thomas, 1984;  Hopkins,  1983)
Saeger,  V.W.   and   E.S.  Tucker.   1973a.   Phthalate  esters  undergo  ready
blodegradatlon.  Plast.  Eng.  (August)   p.  46-49.
04790
IX-31
07/05/91
-------
Saeger,  V.H.  and E.S. Tucker.   1973b,   Blodegradatlon  of phthalate esters.
Technical  Paper   for  Regulatory  Technology  Conference  of  the  Society  of
Plastlclzers.  England Palisades Section.   March  20-22.   p. 105-113.
Saeger, V. and  E.  Tucker.   1976.   Blodegradatlon of phthallc acid esters 1n
river water and activated sludge.   Appl.  Environ.  MUroblol.   31:  29-34.

Saicural, T.,  S.  Wyazawa and T.  Hashimoto.   1978.   Effects of d1-(2-ethyl-
hexyl) phthalate administration on carbohydrate and fatty add metabolism 1n
rat liver.  J. Blochem.  (Tokyo).   83:  313-320.

Sasaki, S.  1978.  The  scientific aspects  of the chemical  substance control
law   In   Japan.    In:   Aquatic  Pollutants:  Transformation   and   Biological
Effects, 0. Hutzlnger,  L.H.  Von  Letyoeld and B.C.J. Zoeteman. Ed.  Pergamon
Press, Oxford,  p.  283-298.

Schmld,   P.   and  C.   Schlatter.   1985.   Excretion   and  metabolism  of
d1(2-ethylhexyl)phthalate In man.   Xenoblotlca.   15: 251-256.

Schultz, C.O. and  R.J.  Rubin.  1973.  Distribution, metabolism and excretion
of  d1-(2-ethylhexyl)  phthalate   1n   the  rat.    Environ.   Health Perspect.
3: 123-129.
Schultz, C.O., R.J. Rubin and G.H. Hutchlns.  1975.  Acute  lung  toxldty and
sudden  death  In  rats  following  Intravenous  administration  of  the plastl-
clzer,  d1-(2-ethylhexyl)  phthalate,   solublUzed  with   Tween   surfactants.
Toxlcol. Appl. Pharmacol.  33:  514-525.
04790
IX-32
07/05/91
-------
Seed,  J.L.    1982.   Mutagenlc  activity  of  phthalate  esters  in  bacterial
liquid suspension assays.  Environ.  Health Perspect.   45:  111-114.

Seth,  F.K.   1982.   Hepatic  effects  of phthalate  esters.   Environ.  Health
Perspec:.  45: 27-34.

Seth.  P.K.,  S.P.  SMvastava,  M.  Mushtag,  O.K.  Agarwal  and  S.V.  Chandra.
1979.  Iffeet of d1-(2-ethylhexyl) phthalate on rat  liver Injured  by  chronic
carbon ietrachlorlde treatment.  Acta Pharmacol. Toxlcol.  44:  161-167.

Seth,  P  K.,  O.K. Agarwal  and S.  Agarwal.   1981.   Effect  of  phthalU  acid
esters 
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Shlota,  K.  and  H.  Nlshlmura.   1982.   TeratogenlcHy  of  d1-{2-ethylhexyl)
phthalate  (DEHP)  and d1-n-butyl  phthalate (DBP)  In  mice.   Environ. Health
Perspect.  45: 65-70.
Short,  R.O.,  E.G.  Robinson,  A.H.  Llngton and  A.E.  Chin.   1987.  Metabolic
and peroxlsome  proliferation  studies  with d1(2-ethylhexyl)phthalate In  rats
and monkeys.  Toxlcol. Ind.  Health.   3(1): 185-195.

Slintion,  V.,  K.  Kauhanen  and  R.  TardUf.   1977.   Mutagenlc  activities  of
chemicals  Identified   In   drinking  water.   Oev.   Toxlcol.   Environ.   Scl.
2: 249-258.

Singh, A.R., W.H.  Lawrence  and  J.  Autlan.  1972.  TeratogenlcHy of phthal-
ate esters In rats.  J. Pharmacol.  Scl.   61:  51-55.

Singh, A.R., W.H.  Lawrence  and  3.  Autlan.  1974.  Mutagenlc and  antlfertll-
Hy   sensitivities   of  mice   to  d1-(2-ethylhexyl)  phthalate   {DEHP)   and
dlmethoxyethyl phthalate (DEMP).  Toxlcol.  Appl.  Pharmacol.   29:  35-46.

S'mgh, A.R., W.H.  Lawrence  and 3. Autlan.  1975.  Maternal-fetal  transfer  of
1
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SJoberg,  P.,  N.6.  Linguist,  G.  Hontln and  L.  Ploen.   1985b.   Effects  of
repeated  intravenous  Infusions  of the  plastldzer  d1-(2-ethylhexyl)  phthal-
ate 1n young male rats.  Arch. Toxlcol.  58: 78-83.

SJoberg,  P.,  U.  Bondesson,  L.  Kjellen,  N.-G.  L1nqu1st,  G.  Montln  and  L.
P15en.    i985c.   Kinetics  of dl-(2-ethylhexyl)  phthalate  In  Immature  and
mature  rats and effect on testls.  Acta Pharmacol. Toxlcol.  56: 30-37.

Sjoberg,   P.,  U.G.   Bondesson,   E.G.   Sedln  and  I.P.  GustaFfson.   1985d.
Exposure    of   newborn    Infants    to  plastlclzers:   plasma   levels   of
d1(2-eth>lhexyl)phthalate  and   mono(2-ethylhexyl)phthalate  during  exchange
 transfusion.   Transfusion.   25:  424-428.

 Sjoberg,   P.,  U.G.  Bondesson,  T.J.  Gray  and L. Ploen.   1986a.  Effects  of
 d1(2-eth;'lhexyl)phthalate and five  of Its metabolites  on rat testls  In  vivo
 and in y__tr£.   Arch.  Toxlcol. 58: 72-77.

 Sjoberg,  P., N.G.  Undqvlst  and L.  Ploem.  1986b.   Age-dependent  response of
 the  rat  testes  to  d1(2-ethylhexyl}phthalate.  Environ.  Health Perspect.   65:
 237-242.

 Smith,  C.C.   1953.   Toxlclty  of  butyl   stearate,   dlbutyl  sebacate,  dlbutyl
 phthalate and methoxyethyl  oleate.  Arch.  Ind.  Hyg. Occup. Med.  7: 310-318.

  Smith, ,i.C.,  E.G.  McOanlel, F.F.  Fan and J.A.  Hasted.  1973.  Zinc: A  trace
  element  essential  1n vitamin A  metabolism.  Science.   181:  954-955.
  04790
                                       IX-35
                                                                       07/05/91
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SMvastava,  S.P.,  O.K. Agarwal  and  P.K.  Seth.   1977.   Effects  of  d1-(2-
ethylhexyl)  phthalate  on  activity  of  succlnk  dehydrogenase and  adenoslne
tMphosphatase  of  some  vital   organs  of  rat.   Toxicology.   7:  163-168.
(CHed In Seth,  1982).

SMvastava, S.P., O.K.  Agarwal,  H.  Mushtag  and  P.K.  Seth.   1978.   Effect of
d1-(2-ethylhexyl) phthalate  (DEHP)  on  chemical  constituents and  enzymatic
activity of rat  liver.   Toxicology.  11: 271-275.  (CHed In Seth,  1982).

Stalling,  D.,  J.W.  Hogan  and  3.L.  Johnson.   1973.   Phthalate ester, resi-
dues - their  metabolism and analysis  1n  fish.   Environ.   Health  Perspect.
3: 159-173.

Stenchever,  M.A.,   M.A.  Allen,  L.   Jeromlnskl   and   R.V.   Peterson.   1976.
Effects  of  b1s-(2-ethylhexyl)  phthalate  on chromosomes  of  human  leukocytes
and human fetal  lung cells.   3.  Pharm. Scl.  65: 1648-1651.

Sugatt,  R.H.,  D.P.   O'Grady,  S.  Banerjee,  P.H.  Howard  and  W.E.  Gledhlll.
1984.  Shake  flask  blodegradatlon of  14  commercial  phthalate esters.   Appl.
Environ. M1crob1ol.   47: 601-606.

Sullivan,  K.F.,  E.L.  Atlas  and C.S.  Glam.   1982.    Adsorption of  phthallc
add esters from seawater.   Environ. Sd. Technol.  16: 428-432.
04790                                IX-36                           07/05/91
-------
Tabak, H.H., S.A.  Quave,  C.I.  Mashnl and E.F. Barth.   1981.   B1odegradab1l-
1ty studies for predicting the environmental  fate  of  organic  priority  pollu-
tants,   in:   Test  protocols   for   Environmental   fate  and  Movement   of
Toxicants.  Proc.  Symp.  Assoc.  of  Official  Analytical  Chemistry, 94th  Ann.
Meeting, Washington, DC.   p.  267-328.

Tanaka,  A., T.  Adachl,   T,  Takahashl  and  T.  Yamaha.   1975.   Biochemical
studies  )n  phthalk esters,   I.  Elimination, distribution and metabolism  of
d1-{2-ethylhexyl) phthalate In rats.  Toxicology.   4:  253-264.

Tanaka,  A., A.  Matsumoto  and  T.  Yamaha.   1978.    Biochemical  studies  on
phthallc  esters.    III.  Metabolism  of  dlbutyl  phthalate  (08P)  In  animals.
Toxicology.  9:109-123.

Taylor,   B.F.,   R.W.  Curry   and   E.F.   Corcoran.   1981.    Potential   for
blodegr^datlon  of  phthallc acid esters 1n  marine regions.    Appl.  Environ.
Mlcroblcl.  42(4): 590-595.

Telrlynck,  O.A.  and F. BelpaUe.  1985.  Disposition of  orally  administered
d1(2-eUylhexy1}phthalate  and   mono   (2-ethylhexyl)phthalate   In   the  rat.
Arch. T>x1col. 57: 226-230.

Thless,   A.H.   and   1.   Flelg.     1979.    Chromosomeh    untersuchungen   be1
MHarbeHern   m1t   Exposition  gegenuber   D1-2-Sthylhexylphthalat   (OOP).
Berlchtlgung  Zbl.  Arbeltsmed.,   Bd.    29,  H.  4,   April   1979,  S.  120.  p.
351-35*.  (Ger.)
04790
IX-37
07/05/91
-------
Th'iess, A.M., A.  Korte  and  H.  Flelg.   1978a.  Untersuchengen zur morbldUat
bel  MHarbeltern  mU   Exposition  gegenuber  D1-2-athylhexylphthalat  {DOPJ.
Vortr. Anl, d. Jahrestg. d. Deutschen Gesellschaft f. Arb. Med.  In Frankfurt
V.  25. - 27.5.78.  p.  137-151.   (Ger.)
Thiess, A.M., R.  Frentzel-Beyme  and  R.  Wleland.   1978b.   Mortality study In
workers exposed  to  d1-(2-ethylhexyl) phthalate  (OOP)  (Ger.).   ITK Moglich-
kerten und  Grenzen  des  Biological Monitoring.  Arbe1tsmed1z1n1sche Probleme
des Dlenstlelstungsqewerbes.  ArbeHsmedlzlnlsches kolloqulum  [Possibilities
and Limits  of Biological Monitoring.   Problems  of  Occupational  Medicine In
Small   Industries.   Colloquium 1n  Occupational  Medicine],  Frankfurt/M.,  May
1978.   Stuttgart, A.W.  Gentner,  p. 155-164.
Thomas,  J.A.  and  M.J.  Thomas.   1984.   Biological  effects  of d1-(2-ethyl-
hexyl) phthalate  and  other phthallc  add  esters.   CRC  Crlt.  Rev. Toxlcol.
13(4): 283-317.
Thomas, J.A., T.D.  Darby,  R.F.  Wallln,  P.J.  Garvln and L. Martls.  1978.  A
review  of  the biological  effects  of  d1-(2-ethylhexyl) phthalate.  Toxlcol.
Appl. Pharmacol.   45: 1-27.

Thomas, 3.A.,  K.A.  Curto  and  M.J. Thomas.   1982.   MEHP/DEHP:  Gonadal  tox-
Iclty and effects on rodent accessory sex  organs.   Environ. Health  Perspect.
45: 85-88.
TcmHa,  I.,  Y. Nakamura,  Y.  Yag1  and  K.  Tutlkawa.  1982a.  Teratogenlclty/
fetotoxIcHy of DEHP 1n mice.  Environ.  Health Perspect.   45:  71-75.
0*790
IX-38
07/05/91
-------
TomUa,  I.,  t.  Nakamura, N.  Aok1  and  N.  !nu1.   1982b.  Mutagenlc/carclno-
genlc po:ent1al  of DEHP and  MEHP.   Environ.  Health Perspect.  45:  119-125.

Tsuchlya, K.  and K. HattoM.   1977.   Chromosomal  study  on  human leukocyte
cultures  treated  wHh  phthallc  add  ester.   HokkaldorHus  Elsel Kenkyusho
Ho.  26: 1H.  (Abstract)

Turnbull, D.  and  J.V.  RodMcks.   1985.   Assessment  of possible carcinogenic
risk to  humans  resulting from exposure to  d1(2-ethylhexyl)phthalate (DEHP).
J. Amer. Coll. Roxlcol.  4:  111-145.

Turner,  J.H.,  O.C.  Petrlcdanl,  M.L.  Crouch and  S.  Henger.    1974.   An
evaluation of  the effects of  dlethylhexyl  phthalate  on mHotlcally capable
cells 1r blood packs.  Transfusion.  14: 560-566.

Tyl, R.rf.,  C.O.  Price,  H.C.  Marr  and  C.A.   Klmroel.   1988.   Developmental
toxUUy evaluation  of dietary. d1(2-ethylhexyl Jphthalate In Fischer 344 rats
and CD-'  mice.  Fund. Appl.  Toxlcol.   10(3): 395-412.

U.S. EPA.   1978.  Chemical  Hazard  Information Profile  Draft  Report:  Alkyl
Phthala:es.   Office of  Toxic  Substances, Washington, DC.

U.S.  El'A.    1980.    Ambient  Water   Quality  Criteria  for Phthalate  Esters.
Prepare! by the Office of Health and  Environmental Assessment, Environmental
Criteria  and  Assessment  Office,  Cincinnati,  OH For  the  Office of  Hater
Regulations    and   Standards,  Washington,  DC.   EPA   440/5-80-067.    NT1S
PB81-117780.
04790
IX-39
07/05/91
-------
U.S. EPA.  1982.  Aquatic  Fate  Process  Data  for  Organic Priority Pollutants.
Office of Water Regulations and Standards, Washington, DC.  EPA 440/4-81-014.

U.S. EPA.   1985.   TSCAPP  —  Toxic  Substance  Control  Act  Plant  Production;
Online.

U.S.  EPA.   1986.   Guidelines  for  Carcinogen  Risk  Assessment.    Federal
Register. 51(185): 33992-34003.
U.S.  EPA.   1987a.   Health and  Environmental  Effects  Profile  for  Phthalic
Acid  Alkyl,  Aryl and  Alkyl/Aryl  Esters.  Prepared  by  Office of  Health  and
Environmental  Assessment,  Environmental  Criteria  and  Assessment  Office,
Cincinnati,  OH  for  the  Office   of  Solid  Waste  and  Emergency  Response,
Washington, DC.  EPA/600/X-87/384.  NTIS PB89-120158/AS.

U.S.  EPA.   1987b.   Health  Effects  Assessment  for  Selected Phthalic  Acid
Esters.   Prepared  by  the  Off1.ce  of  Health  and  Environmental  Assessment,
Environmental  Criteria  and Assessment Office, Cincinnati, OH  for  the Office
of  Emergency and Remedial  Response, Washington,  DC.   EPA/600/8-88/053.   NTIS
P888-178934/AS.
U.S.  EPA.   1991.   Integrated  Risk  Information  System  (IRIS).   Online.
Office  of Health  and  Environmental  Assessment,  Environmental Criteria  and
Assessment Office, Cincinnati, OH.  April.
USITC  (U.S.  International  Trade  Commission).   1983.   Synthetic  Organic
Chemicals.   United  States  Production  and   Sales,   1982.   U.S.  Government
Printing Office, Washington, DC.
04790
IX-40
07/05/91
-------
USITC  (J.S.  International  Trade  Commission).   1985.   Synthetic  Organic
Chemicals.  United  States  Production  and  Sales,   1984.   U.S.   Government
Printing Office. Washington, DC.  p. 165.

von Danlken, A., W.K. Lutz,  R.  Jackh  and C.  Schlatter.   19*84.   Investigation
of  the  potential  for  binding  of  d1-(2-ethy1hexy1)  phathalate  (DEHP)  and
d1-(2-etiyhexyl) adlpate  (DEHA)  to  liver ONA In vivo.  Toxlcol.  Appl.  Phar-
macol.   ?3: 373-387.

Waddell,  W.H.,  C.  Marlowe,  O.E.  M1r1po1 and  P.J.  Garvln.   1977.  The  dls-
trlbutloi  In  mice  of  Intravenously  administered  plasma  solutions of  14C-
d1-(2-et lylhexyl)  phthalate  determined   by   whole-body   autoradlography.
ToxUol. Appl. Pharmacol.  39: 339-353.

Wallln,  *.F., B. Klamer,  R.W.  Nlcora  and C.R.  Thompson.   1974.   Q1-(2-ethyl-
hexyl)  phthalate (DEHP)  metabolism 1n   animals  and  post-transfusion  tissue
levels In man.  Bull. Parenteral Drug Assoc.   28:  278-287.

Walseth,  F.,  R.  Toftgard and  O.G.  NHsen.   1982.    Phthalate  esters.   I:
Effects on cytochrome P-450  mediated  metabolism In rat  liver and  lung,  serum
enzymatic activities  and serum protein levels.   Arch.  Toxlcol.   50:  1-10.
Ward,  J.I.,  3.M.  R1ce,  D. Creasla,  et al.   1983.   Dissimilar  patterns  of
promotlor by d1-(2-ethylhexyl)  phthalate  and  phenobarbHal  of  hepatocellular
neoplaslt     Initiated     by    dlethylnltrosamlne
Cardnogcnesls.  4(8): 1021-1029.
                  1n
B6C3F
                             1
mice.
04790
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Ward,  J.M.,  B.A.  Dlwan,  H. Ohshlma,  H.  Hu,  H.H.  Schuller  and  3.M.  Rice.
1986.    Tumor-Initiating   and  promoting   activities   of   d1(2-ethylhexyl)
phlhalate in. vWo and \n vitro.   Environ.  Health Perspect.   65:  279-291.

Warren,  3.R.,  N.D.  Lalwanl  and  J.K.  Reddy.   1982.   Phthalate  esters as
peroxlsome prollferator carcinogens.   Environ. Health  Perspect.  45:  35-40.

Watts,  P.   1985.   01-2-ethylhexylphthalate  metabolism In  man.   Food  Chem.
Toxlcol.  23: 1023.

Weast,  R.C.,  Ed.  1983.   Handbook  of Chemistry  and  Physics, 63rd  ed.   CRC
Prsss, Inc., Boca Raton, FL.

Wllbourn, 3. and R. Montesano.   1982.  An  overview of phthalate ester  car-
clnogenlclty  testing  results:  The  past.   Environ.   Health  Perspect.   45:
127-128.

Wlldbrett, G.  1973.   Diffusion of phthallc  add esters  from PVC milk  tub-
Ing.  Environ.  Health Perspect.   3:  29-35.

Williams, O.T.  and  8.J.  BlanchHeld.  1975.   The  retention,  distribution,
excretion and  metabolism  of dlbutyl  phthalate-7-l«C  In  the rat.   a.  Agrlc.
Food Chem.  23: 854-857.  (Cited In  U.S.  EPA, 1980;  Gangolll, 1982).
Williams, G.M., H. Maruyama and T.  Tanaka.   1987.   Lack  of  rapid Initiating,
promoting or  sequential  syncardnogenlc  effects  of d1(2-ethylhexyl)phthalate
In rat liver carclnogenesls.  Carclnogenesls.  8(7): 875-880.
04790
IX-42
07/05/91

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Wolfe, N l., W.C.  Steen  and  L.A. Burns.  1980.  Phthalate  ester  hydrolysis:
Linear f-ee  energy relationships.   Chemosphcre.  9: 403-408.   EPA  600/J-80-
016.

YanagHa   T.,  S.  Kuzuhara,  N.  Enomoto,  T.  Shlmada and  M.  Sugano.   1979.
Effects  of  d1-(2-ethylhexyl)  phthalate on  the content  and  composition  of
hepatic  mltochondrlal  and  mlcrosomal  phosphollplds  1n  the  rat.   Blochem.
Pharmacol.  28: 3115-3122.

Yoshlkawi,  K.,  A.  Tanaka,  T.  Yamaha  and  H.  Kurata.   1983.  Mutagenldty
study of  nine  monoankyl  phthalates  and a dlalkyl phthalate using Salmonella
typhlmurlum and EscherlcMa coll. Food Chem.  Toxlcol.  21(2):  221-223.

Yurchenko,  V.V.   1977.   Cytogenetlc  study  of mutagenlc properties  common  to
the  rep^^lants  dimethyl  phthalate   and  N,N-d1ethylam1de  of  phenoxyacetU
add.  Farmakol. Tokslkol. (Moscow).   4: 454-457.   (Rus.)

Yurchenko,  V.V.  and S.  Glelberman.    1980.   Study   of  long-term  effects  of
repellart  use.  Part III.  Study of mutagenlc properties  of dimethyl  phthal-
ate  and  phenoxyacetlc  add  N.N-dlethylamlde by dominant lethal  mutations.
Med. Parazltol. Parlzlt.  Boleanl.  49: 58-61.  (Rus.)

Zelger, E.,  S.  Haworth,  W.  Speck and  V. Mortleraans.   1982.   Phthalate  ester
testing  In the National  Toxicology Program's environmental mutagenesls  test
developnent program.  Environ.  Health  Perspect.   45:  99-101.
04790
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