EPA-540/1-86-025
                                             Office of Emergency and
                                             Remedial Response
                                             Washington DC 20460
Off'ce of Research and Development
Office of Health and Environmental
Assessment
Environmental Criteria and
Assessment Office
Cincinnati OH 45268
                       Superfund
&EPA
                        HEALTH EFFECTS ASSESSMENT
                        FOR  COPPER

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                                           EPA/540/1-86-025
                                           September 1984
       HEALTH EFFECTS  ASSESSMENT
                FOR  COPPER
    U.S. Environmental Protection  Agency
     Office of Research and Development
Office of Health  and  Environmental Assessment
Environmental Criteria and Assessment  Office
            Cincinnati,  OH  45268
    U.S. Environmental Protection  Agency
  Office of Emergency  and Remedial Response
Office of Solid Waste and Emergency  Response
            Washington,  DC  20460

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                                  DISCLAIMER

    This  report  has  been  funded  wholly  or  In  part  by  the  United  States
Environmental  Protection  Agency under  Contract  No.  68-03-3112  to  Syracuse
Research Corporation.  It has been  subject  to the Agency's peer and adminis-
trative review, and  1t has  been  approved  for  publication as an EPA document.
Mention of  trade  names or  commercial  products  does  not  constitute  endorse-
ment or recommendation for use.
                                      11

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                                    PREFACE


    This report  summarizes  and evaluates Information relevant  to  a prelimi-
nary  Interim  assessment  of  adverse health  effects  associated with  copper.
All  estimates  of acceptable  Intakes and  carcinogenic  potency presented  1n
this  document  should  be  considered   as  preliminary  and  reflect  limited
resources   allocated   to   this   project.     Pertinent   toxlcologlc   and
environmental  data  were  located  through  on-line literature searches  of  the
Chemical  Abstracts,  TOXLINE,   CANCERLINE   and   the  CHEMFATE/DATALOG  data
bases.  The basic literature searched supporting this document  1s  current  up
to September, 1984.   Secondary sources  of Information have  also  been  relied
upon  In the  preparation of  this  report  and  represent  large-scale  health
assessment  efforts   that entail   extensive   peer  and  Agency  review.   The
following Office  of  Health  and  Environmental  Assessment (OHEA)  source  has
been extensively utilized:


    U.S.  EPA.    1980a.    Ambient  Water  Quality  Criteria Document  for
    Copper.   Environmental  Criteria and Assessment  Office,  Cincinnati,
    OH.   EPA  440/5-80-036.    NTIS  PB  81-117475.  (Cited In U.S.  EPA,
    1985)

    U.S.  EPA.   1983a.   Technical  Support  Document  on  the Ranking  of
    Hazardous  Chemicals  Based  on  Cardnogenldty.   Prepared  by  the
    Carcinogen Assessment Group,  OHEA,  Washington, DC for the  Office of
    Emergency and Remedial Response, Washington,  DC.

    U.S.  EPA.    1985.   Drinking  Water  Criteria  Document  on  Copper.
    Prepared  by  the  Environmental  Criteria  and  Assessment  Office,
    Cincinnati,  OH  OHEA  for the  Office of  Drinking Water,  Washington,
    DC.  Final Draft.
    The Intent 1n these assessments  1s  to  suggest  acceptable exposure levels
whenever sufficient data  were  available.   Values were not  derived  or larger
uncertainty  factors  were  employed  when the  variable data  were limited  1n
scope tending  to  generate conservative  (I.e.,  protective)  estimates.   Never-
theless, the  Interim  values  presented reflect the relative  degree  of hazard
associated with exposure or risk to the chemlcal(s) addressed.

    Whenever possible, two categories of values  have  been  estimated for  sys-
temic toxicants (toxicants for which  cancer  Is  not the endpolnt of  concern).
The  first,  the AIS  or  acceptable  Intake  subchronlc, 1s  an estimate of  an
exposure  level  that  would not  be  expected  to cause  adverse  effects  when
exposure occurs during  a  limited  time  Interval  (I.e., for  an  Interval  that
does not  constitute  a  significant  portion of  the Hfespan).   This  type  of
exposure estimate  has not been  extensively used  or  rigorously  defined,  as
previous  risk  assessment  efforts  have  been  primarily  directed  towards
exposures from toxicants  In  ambient air or water  where  lifetime exposure  1s
assumed.  Animal  data  used  for  AIS estimates  generally  Include  exposures
with durations of  30-90  days.   Subchronlc  human data are  rarely available.
Reported exposures are  usually  from chronic  occupational  exposure situations
or from reports of acute accidental exposure.
                                      111

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    The AIC,  acceptable  Intake  chronic,  is  similar 1n  concept  to  the  ADI
(acceptable  daily  intake).   It  is  an  estimate  of an  exposure  level  that
would not  be expected  to cause  adverse effects  when  exposure occurs  for  a
significant portion  of  the lifespan  [see  U.S.  EPA  (1980b)  for a discussion
of  this  concept].   The  AIC  is  route specific   and  estimates   acceptable
exposure  for  a given  route with  the  Implicit  assumption  that  exposure  by
other routes is insignificant.

    Composite  scores  (CSs)  for  noncarcinogens  have  also  been  calculated
where data  permitted.   These  values  are used for  ranking  reportable quanti-
ties; the methodology for their development is explained  in U.S.  EPA (1983b).

    For compounds for which there  is  sufficient  evidence  of  carcinogenicity,
AIS  and AIC values  are  not derived.   For a  discussion  of risk assessment
methodology  for  carcinogens  refer  to  U.S. EPA  (1980b).   Since  cancer  is  a
process that  is  not  characterized by  a threshold,  any exposure  contributes
an Increment of risk.   Consequently,  derivation of  AIS and  AIC values would
be inappropriate.   For  carcinogens,  q-|*s  have been computed  based  on  oral
and Inhalation data 1f available.
                                      1v

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                                   ABSTRACT


In  order  to  place  the  risk  assessment  evaluation  1n  proper  context,  the
reader  1s  referred  to  the  preface of  this document.  The  preface outlines
limitations  applicable  to   all  documents  of   this  series  as  well  as  the
appropriate Interpretation and use of the quantitative estimates.

    Copper Is an essential trace  element.   In  Individuals  with normal copper
metabolism and  normal levels  of  G6PD,  there  seems  to be a  wide separation
between required levels and  toxic  levels.   The present document reflects the
estimate of 2.6 mg/day  for both  the  AIS and AIC.   This Is  the dally exposure
level suggested by U.S. EPA  (1985) and  1s  based on human GI  symptoms follow-
ing acute  exposure.   This value  is  also 1n good  agreement  with the limited
animal  data.   A CS  of  19 was  estimated for elevated  serum  AST activity and
jaundice In pigs fed high levels of copper sulfate.

    No  good   quantitative animal  data  exist   for  Inhalation  exposure  and
effects of  copper.   For  this  reason  the  TLV values, 0.2  mg/m3  for  fumes
and  1.0 mg/m3  for  dusts  and mists, were  used to estimate  Inhalation  AICs.
The ACGIH  (1983)  based  these levels  on extensive  Industrial  experience with
copper  In Great Britain.  The  estimated  AICs are:   0.14 mg copper vapor/day;
0.71  mg copper mist  or  dust/day.  These  suggestions  should  be reviewed  as
more complete data become available.

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                               ACKNOWLEDGEMENTS
    The  Initial  draft  of  this  report  was  prepared  by  Syracuse  Research
Corporation  under-  Contract No.  68-03-3112  for  EPA's  Environmental  Criteria
and  Assessment  Office,  Cincinnati,   OH.   Dr.  Christopher  DeRosa and  Karen
Blackburn were the Technical  Project  Monitors  and  Helen Ball  was >,the Project
Officer.  The final documents  1n  this series  were  prepared for the Office of
Emergency and Remedial Response, Washington, DC.

    Scientists from  the  following U.S. EPA offices  provided  review comments
for this document series:

         Environmental Criteria and Assessment Office, Cincinnati, OH
         Carcinogen Assessment Group
         Office of A1r Quality Planning and Standards
         Office of Solid Waste
         Office of Toxic Substances
         Office of Drinking Water

Editorial review for the document series was provided by:

    Judith Olsen and Erma Durden
    Environmental Criteria and Assessment Office
    Cincinnati, OH

Technical support services for the document series  was provided by:

    Bette Zwayer, Pat Daunt, Karen Mann and Jacky Bohanon
    Environmental Criteria and Assessment Office
    Cincinnati, OH
                                      v1

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TABLE OF CONTENTS
1.
2.


3.








4.




5.
6.









7.
,PPE
ENVIRONMENTAL CHEMISTRY AND FATE 	
ABSORPTION FACTORS IN HUMANS AND EXPERIMENTAL ANIMALS . . .
2.1. ORAL 	
2.2. INHALATION 	
TOXICITY IN HUMANS AND EXPERIMENTAL ANIMALS 	
3.1. SUBCHRONIC 	
3.1.1. Oral 	
3.1.2. Inhalation 	
3.2. CHRONIC 	
3.3. TERATOGENICITY AND OTHER REPRODUCTIVE EFFECTS. . . .
3.3.1. Oral 	
3.3.2. Inhalation 	
3.4. TOXICANT INTERACTIONS 	
CARCINOGENICITY 	
4.1. HUMAN DATA 	
4.2. BIOASSAYS 	
4.3. OTHER RELEVANT DATA 	
4.4. WEIGHT OF EVIDENCE 	
REGULATORY STANDARDS AND CRITERIA 	
RISK ASSESSMENT 	
6.1. ACCEPTABLE INTAKE SUBCHRONIC (AIS) 	
6.1.1. Oral 	
6.1.2. Inhalation 	
6.2. ACCEPTABLE INTAKE CHRONIC (AIC) 	
6.2.1. Oral 	
6.2.2. Inhalation 	
6.3. CARCINOGENIC POTENCY (q-|*) 	
6.3.1. Oral 	
6.3.2. Inhalation 	
REFERENCES 	
NDIX: Summary Table for Copper 	
Page
1
3
. . . 3
4
, , , 5
5
. . . 5
, , , 10
. . . 10
. . . 11
. . . 11
12
. . . 12
, . . 14
. . . 14
. . . 14
. . . 15
, , , 17
18
. . . 20
, . . 20
. . . 20
. . . 22
. . . 22
. . . 22
. . . 23
. . . 24
. . . 24
25
. . . 26
. . J34

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                               LIST OF TABLES

No.                               Title                                Page

3-1     Effects of Copper Sulfate*5H20 Administered
        1n Corn-Soy Diet	    7

4-1     Tumorlgenlclty of Some Copper Compounds 	   16

5-1     Current Regulatory Standards and Criteria 	   19

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                             LIST  OF  ABBREVIATIONS
ADI                     Acceptable dally Intake
AIC                     Acceptable Intake chronic
AIS                     Acceptable Intake subchronlc
AST                     Aspartate transamlnase
BCF                     Bloconcentration factor
bw                      Body weight
CAS                     Chemical Abstracts Service
CS                      Composite score
CNS                     Central nervous system
DNA                     Deoxyr1bonucle1c acid
GI                      Gastrointestinal
GRAS                    Generally regarded as safe
G6PD                    Glucose-6-phosphate dehydrogenase
MED                     Minimum effective dose
NOAEL                   No-observed-adverse-effect level
ppm                     Parts per million
RVj                     Dose-rating value
RVe                     Effect-rating value
TLV                     Threshold limit value
TWA                     Time-weighted average
                                      1x

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                     1.   ENVIRONMENTAL CHEMISTRY AND FATE
    Copper  1s  a  metal  belonging  to  the  First Transitional  Series  of  the
periodic  table.   Elemental copper  has a  CAS  number  of  7440-50-8.   Copper
occurs 1n nature as the elemental metal  (zero  valence), and  In the +1  and +2
valence  states.   In addition  to a  variety of  Inorganic compounds,  copper
forms  a  number  of  compounds  with  organic  Ugands.    Both  organic   and
Inorganic copper  compounds  have a variety  of  uses  (Kust, 1979).   Most-  Cu
compounds are not stable 1n the  environment,  particularly In  the presence of
water  or moisture  and  air,  and tend  to  change  to  the  stable  Cu+   state
(Kust, 1979).
    In  the  atmosphere,   copper  1s  present as  dusts  and  fumes from  copper
smelting  Industries,  Iron  and  steel  Industries,  coal burning  power  plants
and  other  miscellaneous   fabricating  operations   Involving   copper   (NAS,
1977).   The atmospheric  fate  of  copper  has  not  been  studied  comprehen-
sively.   Any chemical  Interaction  of  copper  compounds 1n the  atmosphere  1s
likely to result  1n  spedatlon (I.e., conversion of copper compounds  Into a
stable  species  such as   CuO),  not  1n  Us  direct removal   through  decomposi-
tion  as   frequently  occurs with  organic compounds.   The principal  removal
mechanisms  for atmospheric  copper are probably wet  and dry deposition.   The
atmospheric   half-life  for  the  physical  removal  mechanism  1s  expected  to
depend on the  particle  size and particle density  of atmospheric copper.   No
estimate for the atmospheric half-life of copper Is available.
    The  aquatic  fate  of copper  has   been  studied  more extensively than  Us
atmospheric  fate (Callahan et  al., 1979).   The  two  processes  that  are  likely
to dominate  the  fate  of  copper 1n aquatic  media are chemical  spedatlon  and
sorptlon  (Callahan  et  al.,  1979).   The nature  of chemical  spedatlon  of
copper  in aquatic media 1s  determined by  the  oxidation-reduction  potential
                                     -1-

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of  the  particular  copper  compound  and  the  pH  of  the  aquatic media.   In
aquatic media  of  pH <7, copper  may  exist  1n  Cu*  form,  whereas at  pH >7,
copper may exist as  the  carbonate  complex.   In  polluted water bodies, copper
may  form  complexes  with organic  material  1n  the  water.   Various  sorptlon
processes  reduce   the  level   of  1on1c  state  carbonate  complex  or  organic
complex of copper  from aquatic media.   Sorptlon onto  clay materials,  hydrous
Iron,  manganese  oxides  and  organic  material   1s  the primary  controlling
factor  (Callahan  et  al., 1979).    In  organically rich  sediments, the sorbed
and  precipitated  copper  may  become redlssolved  through complexatlon  and  may
persist 1n the  water for a long time.  No  estimate of  the aquatic  half-life
of copper 1s  available 1n the literature.
    The fate  of copper   1n soil  has  been studied Inadequately;  however,  the
fate may  depend upon the pH of  the  soil,  Us moisture content and  Its clay
and organic matter content (NAS, 1977).   In  acidic  soils,  copper  may be more
soluble, which  would enhance  Us  mobility  (NAS, 1977);  the reverse  may  be
true 1n basic  soils.  Soils  rich 1n organic  matter may enhance  the  mobility
of copper through  complexatlon.  Both clay  and  organic  matter may facilitate
the  sorptlon  of copper   in  soil, however,  and  may  retard  Its  Teachability.
Soils with suitable  moisture  content may enhance the microorganism  activity
and  the partial  removal of  copper   through  uptake  by microorganisms.   No
estimate of  the half-life  of copper  in soils  is available;  however,  copper
is expected to  be  leached more readily  from acidic  and  sandy soils  than from
basic soils containing a  higher  percentage of clay and/or  organic  matter.
    The BCFs  for copper   1n aquatic organisms  have been  determined by several
investigators and  have  been  found  to  vary  from 12  for an  alga,  Scenedesmus
quadricarda.  to 30,000 for  molluscs (Callahan et al.,  1979).
                                     -2-

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           2.   ABSORPTION  FACTORS  IN  HUMANS  AND  EXPERIMENTAL  MAMMALS
2.1.   ORAL
    According  to Schroeder  et  al.  (1966), humans  Ingest  an  average  of 2.5-5
mg  of  copper/day  from dietary  sources.   These  authors  estimated   that  61
absorption of  -3.2  and 0.2 mg  of  copper occurs from food and  fluid Intake,
respectively.  The actual quantities  of  copper  absorbed  depend  on geography,
climate, soil chemistry, diet,  water  softness and pH.
    Weber  et  al.   (1969)   administered   64Cu   as  copper  acetate  to  seven
fasted human  subjects  without  liver  damage.  Because of  the short half-life
of  64Cu  (12.8 hours),  labeled  (9SZr)   zirconium oxalate  was  given  as  a
non-absorbable stool marker  to  enable location of the copper  acetate bolus.
The radlonucHdes were counted dally  for  4 days  1n a whole body scintilla-
tion counter,  and GI movement  of  the administered  bolus  was  monitored with a
scintillation  camera.   Radioactive copper  1n  blood was  determined hourly for
6 hours and 1n urine and stoo.ls dally.
    Absorption  of  64Cu  appeared   to  be  diphasic.   Maximum absorption  from
the stomach and duodenum  occurred  within 1  hour of administration.  A second
and slower absorption  phase was observed >3.5 hours  post-administration.  At
2 hours  post-administration the 64Cu  acetate  bolus  had  left  the stomach of
the subjects  and  was  located  1n  the small  Intestine;  3 hours  later  H was
located  in  the  terminal   Heocecal  region  and  proximal   large  Intestine.
Average  net  absorption of  64Cu was  -60%, with a  range  of  15-97% (Weber et
al., 1969).   Evans  (1973)  stated  that,  1n  mammals,  alimentary  absorption of
copper occurs  only from the upper  GI tract  and that  the extent of absorption
may be Influenced by competition  of  other  metals  for metallothloneln binding
sites  (necessary for active transport  of copper),  levels  of  dietary  protein,
kinds  and amounts of anlons present and  the level of  dietary ascorbic add.
                                     -3-

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2.2.   INHALATION
    Quantitative  data   regarding   absorption  of   copper   from  Inhalation
exposure could not be located  1n the  available  literature;  however,  presump-
tive  evidence  has been  located.    Vlllar  (1974)  observed  copper-containing
granulomas  1n  the  lung,  liver  and  kidney  upon  necropsy  of an  Individual
occupatlonally exposed to  Bordeaux  mixture (an aqueous  solution  of  lime and
1-2%  copper  sulfate)  used  1n  spraying  vineyards.   Plmental  and  Menezes
(1975)  noted  copper-containing liver  granulomas  1n  three  patients  who had
used  Bordeaux  mixture  while spraying  vineyards to  prevent  mildew.   Gleason
(1968)  reported  symptoms of "metal  fume fever" (general discomfort,  fever,
chills, stuffiness of the  head) In  three workers  exposed to fine copper dust
at  concentrations of  0.03-0.12 mg/m3.   Installation  of an exhaust  fan that
reduced air levels to <0.008 mg/m3  promptly alleviated these symptoms.
    Batsura (1969) exposed rats (strain,  sex  and  number not  specified) for
15,  30,  45,   60  or  180  minutes   to  50-80  mg copper  ox1de/m3.   Electron
microscopy  showed  that  copper   absorption  had  occurred  1n  rats  exposed for
180 minutes.   Copper oxide particles  had penetrated  the epithelial  cells  of
the pulmonary alveoli and were  found  1n  plasma  6 hours after exposure began.
Particles were also found 1n the proximal convoluted tubules of the kidney.

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                3.  TOXICITY IN HUMANS AND EXPERIMENTAL ANIMALS
3.1.   SUBCHRONIC
3.1.1.    Oral  Exposure.   There  are  a  number   of   acute/subchronlc  case
reports of accidental human exposure.  Only  those  which  provided  data suffi-
cient to estimate exposure levels are summarized here.
    Chattanl   et  al.  (1965)  evaluated  clinical data from  53  patients  who
Ingested copper sulfate 1n suicide attempts.  The  amounts  of  copper  Ingested
ranged   from  0.25-7.6  g of  copper.   Five  patients  died  and the  survivors
exhibited  a   variety  of   symptoms  most  prevalent   of  which  were  nausea,
vomiting and  epigastric pain.
    Semple et  al.  (1960)  reported an outbreak  of  gastroenteritis  affecting
18/50  workmen  following   1ngest1on  of  copper  sulfate  contaminated  water.
Symptoms  Included   dizziness,   headache,  diarrhea,   vomiting  and  abdominal
pain.  Later  analysis  of  the water  source  showed  copper  levels  of  >44  ppm.
Assuming each  man  drank 1 cup  (0.23 8.)  the  estimated dose was  0.143 mg/kg
copper  (U.S.  EPA, 1985).
    Nicholas  and Brlst (1968)  reported  a similar  Incidence.   In  this  case
9/20 had diarrhea,  6/20 vomiting and  9/20 nausea.   A  sample of  tea  showed 30
ppm copper.  U.S. EPA (1985)  estimated the dose  was >0.1  mg/kg.
    Wyllle (1957)  reported an  outbreak  of  copper poisoning  due to  copper
leaching from a  cocktail  shaker.   Amounts of copper  Ingested were  estimated
to  be   5.3-32  mg  copper.   Of   the  women  exposed 10/15  reported  symptoms
Including  weakness,   abdominal   cramps,   headaches,  nausea,   dizziness   and
vomiting.
    Little Information  exists  concerning subchronlc   toxldty  of copper  In
the usual  laboratory  species.   Howell (1959) maintained rats on diets  con-
taining 5000  ppm copper acetate for  16  months.  Assuming  that rats  consume
                                     -5-

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food equivalent  to 5%  of  their bw/day,  these  rats  consumed -250  mg copper



acetate  or  -80 mg  Cu/kg  bw/day.   No  criteria  of toxldty  were  mentioned.



Liver and  kidney  were found  to accumulate copper heavily,  but  no  accumula-



tion was found 1n the cornea or brain.



    Dietary  levels  of -200  ppm copper  have  long been  used as growth  pro-



moters  In the production of  market  hogs.   Kline  et al. (1971) exposed groups



of 12 Hampshire and Yorkshire  pigs  weighing  an  average of 22.2 kg to dietary



levels  of  0, 150,  200  or  250  ppm copper sulfate for 88 days  (Table 3-1).



Accelerated  rate  of  weight  gain  and elevated  levels  of liver copper  were



demonstrated  at  all  treatment  levels.   Hepatic  copper  levels   linearly



(p<0.05) correlated  with  dose.   Depressed growth rate and  blood  hemoglobin



concentration were  observed  1n pigs  fed a diet  containing 500 ppm copper



sulfate for 61 days.



    Suttle  and  Mills  (1966a)   added  750 ppm  of  basic  copper  carbonate



(CuCO/,'Cu(OH)9'H_0)   to   the   cornmeal   diets   of  weanling   female  Large
     
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                                                 TABLE 3-1

                        Effects of Copper Sulfate«5H20 Administered 1n Corn-Soy Diet*
Species/Strain
Pig/Hampshire
and Yorkshire
Pig/Hampshire
Sex/No.
NR/8
NR/8
Average
Body Weight
(kg)
23.6
23.6
Dose
250 ppm diet
3.2 mg Cu+2/kg/day
500 ppm diet
Duration
(days)
61
61
Effects
accelerated growth with
less feed
reduced growth and
and Yorkshire
Pig/Hampshire      NR/12
and Yorkshire
Pig/Hampshire      NR/12
and Yorkshire

Pig/Hampshire      NR/12
and Yorkshire

Pig/Hampshire      NR/12
and Yorkshire
22.2



22.2


22.2


22.2
            5.5  mg Cut2/kg/day
0 supplemental
Cu
150 ppm diet supplement;
1.8 mg Cu*2/kg/day

200 ppm diet supplement;
2.5 mg Cu*2/kg/day

250 ppm diet supplement;
2.9 mg Cu*2/kg/day
         hemoglobin levels.
         Increased liver copper
         concentrations

88       normal hemoglobin,
         hematocrlt and liver
         copper levels

88       accelerated weight gain
88       accelerated weight gain
88       accelerated weight gain
'Source:  Kline et al., 1971

NR = Not reported

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appears that  the Increases  1n  AST levels,  jaundice and anemia  principally
reflected  liver  damage,  since  the  postmortem  examinations  revealed  gross
hepatic  degenerative  changes,  hlstologlc  centrllobular necrosis  and  bile
canallcull  disruption,  and  there  was no  evidence of  Increased  erythrocyte
fragility.  The  addition of  zinc  or  Iron  eliminated the jaundice  and  pro-
duced  serum copper  and AST concentrations similar  to  control  levels after 4
weeks, but only supplemental Iron afforded protection against the anemia.
    In a  second  study  (Suttle  and  Mills,  1966b),  weanling female Large White
pigs  (6/group) were  maintained  on  cornmeal  diets  that contained either soya-
bean  meal,  dried skim  milk  or  whlteHsh  meal as protein supplements with 600
ppm  basic copper carbonate  for  48 days  (6.4,  11.0  and  15.4  mg Cu "Vkg/day,
respectively).   The  soya-bean  meal diet  was  similar  to  that  used previously
1n the 750  ppm  study (Suttle and Mills,  1966a).  Results for control experi-
ments  (no supplemental  copper) were not reported,  but  1t was concluded that
600  ppm  copper  carbonate was  only marginally  toxic  (causing   slight growth
depression  and  a temporary  Increase  1n  serum copper,  negligible  effect  on
AST  levels  and  gross evidence  of  toxicosis  and jaundice 1n only  1  of the 6
pigs).  The effect  of  the dried skim  milk diet with 600 ppm copper carbonate
on the pigs was  also unremarkable, but  the Introduction of whlteflsh meal to
the   diet  reportedly  caused  a  moderately  severe  toxicosis  (marked growth
retardation,  elevated  serum Cu and AST  levels, visible loss of condition and
jaundice  1n 4 of the  6  pigs).  The  greater toxlclty of  the  whlteflsh meal
diet  was  attributed  to  a  higher  calcium level,  which  presumably adversely
Influenced   zinc  availability.    Anemia   developed   gradually  throughout
exposure  1n pigs maintained  on  all the diets.
     In a  related experiment,  250  or  425  ppm of  copper sulfate pentahydrate
(CuSO.-5H?0)  was added  to  the  high calcium cornmeal  whlteflsh  diet  and
                                      -8-

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fed  to  weanling female Large  White  pigs (6/group),  creating  optimal  condi-
tions  for  the  development  of  copper  toxicity  (Suttle  and  Mills,  1966b).
Administration  of  425 ppm  copper  sulfate  1n  the  diet caused  severe  growth
depression after 14 days, when  "severe  toxicosis"  became  evident; three pigs
were slaughtered on  the  47th day and another one  on  the  60th  day to prevent
unnecessary  suffering.   Autopsy revealed  generalized  jaundice,  hypertrophy
and  cirrhosis  of  the liver, and  GI  hemorrhages.   Food consumption  and food
conversion efficiency  data were not  provided  for   these  animals; therefore,
the  daily  intake  of  Cu  *  could not  be estimated.   When pigs were fed 250
ppm  in  the  whHeflsh  meal  diet  for 79  days  (2.6  mg Cu   /kg/day),  slight
weight  gain  was noted over  the first 30 days of  treatment,  but  both  serum
AST  and copper  concentrations  were significantly greater  than  control  values
at  the  46th  day,  when  three  of  the  six   pigs showed  signs of  jaundice.
Concentrations  of  copper  in the liver were  significantly Increased  relative
to  unexposed controls  after  79  days  of  treatment,  but hemoglobin  levels
remained normal.
     It  is  well  recognized  that sheep are  especially sensitive  to  toxicity
due  to  copper.  Underwood  (1977)  described  chronic copper toxicity  1n  sheep
grazing pastures  in  Australia  in  which the  content of  copper  In  soil  and
forages  was  abnormally  high  and/or   forage   levels  of   molybdenum   were
unusually low.  Liver damage from grazing Heliotropium  europaeum  exacerbated
the  toxicity of high  levels  of  copper.   In  ruminants,  dietary levels  of
other  trace  elements,  such as  zinc, affect  the  toxicity  of copper  (MAS,
1977).   Ruminants,  however,  are particularly unsuitable  for  use  as a  basis
for  human  risk assessment,  and  further discussion  of  copper  toxicity  in
these species will  not be included  in this document.
                                     -9-

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3.1.2.   Inhalation.   Pertinent   data  regarding  the  subchronlc  Inhalation
toxlclty  of  copper  1n  laboratory  species  could  not   be  located  In  the
available literature.
3.2.   CHRONIC
    Pertinent  data  regarding chronic   exposure   of  laboratory  animals  to
copper could  not  be located  In  the available literature.   The  most  studied
form of chronic copper toxlclty  1n  humans  Is  Wilson's disease,  or hepatolen-
tlcular  degeneration  (Williams,  1982).   An   Inherited   autosomal  recessive
disorder  of  copper metabolism,  this  disease  1s characterized  by abnormally
low plasma  levels  of ceruloplasmln,  Increased  plasma copper levels  and In-
creased copper  deposition  In  liver, brain, kidneys and  cornea  (Schroeder  et
al.,  1966;  Evans,  1973).   Copper  may  accumulate  In  the liver  of  affected
persons to a  level  -20-fold that  of unaffected  Individuals.   This high level
of hepatic copper  destroys the hepatocytes, resulting In a release of copper
Into the  general circulation.  This  released  copper has  many adverse effects
Including damage  to erythrocytes,  kidneys, corneas and  the  CNS (Schelnberg
and Sternlleb,  1969).  Symptoms  Include  tremors  to drooling, Incoordlnatlon,
seizures, behavioral abnormalities, anemia, jaundice and eventually death.
    Another  manifestation  of  chronic copper  toxlclty 1n man Is  the occur-
rence of  "vineyard  sprayer's  lung"  resulting  from  exposure to copper  sulfate
1n Bordeaux mixture, used  to  control  mildew 1n  grapes (Plmental  and Marques,
1969).   Vlllar  (1974)  further described  the  symptoms of vineyard sprayer's
disease 1n 14 male  patients and  1  female patient  with a prolonged history of
Intermittent  (~3  months/year) exposure  to  Bordeaux mixture.   Dyspnea,  weak-
ness,  anorexia, weight  loss,  radlographlc  opacities  and  the   presence  of
copper  In the lungs were  noted.   Eventually  the  pulmonary  opacities showed
                                     -10-

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regression followed  by calcification.   Later,  Plmental  and Menezes  (1975)
demonstrated   copper-containing   granulomas   In  vineyards   sprayer's   lung
patients exposed for 3-15  years to Bordeaux mixture.
    The  occurrence  of  metal   fume  fever  In  workmen  Involved   1n  polishing
copper  plates has  been discussed  1n  Section  3.1  (Gleason,  1968).   General-
ly, air  samples 1n  the workplace contained  0.30-0.75 mg Cu/m3.   A breath-
Ing  zone air  sample  of  the  polishing  wheel  operator  contained   0.120  mg
Cu/m3.   Installation  of a  ventilation  system  reduced  copper levels  1n  air
to <0.008 mg/m3  and resulted 1n total abatement of symptoms.
3.3.   TERATOGENICITY AND OTHER REPRODUCTIVE EFFECTS
3.3.1.   Oral.  Copper  deficiency  has  been associated with  neural  degenera-
tion,  reduced  growth,  skeletal  malformations  and  cardiovascular  lesions  1n
lambs,  goats, rats,  guinea  pigs, dogs and chickens  (Hurley  and  Keen, -1979).
Ferm  and Hanlon (1974) and DICarlo  (1980)  showed  conclusively  that paren-
teral  administration  of   solutions   of   copper  sulfate  or  copper  citrate
produced  terata 1n  golden hamsters.   Ferm  and  Hanlon  (1974)  demonstrated
that  the chelated   (citrate)  form was a  far  more potent teratogen  than  the
Inorganic (sulfate) form.
    The  teratogenldty of  copper  sulfate administered  In  the   diets  to  two
strains  of  mice was  studied  by Lecyk (1980).  Groups  of 7-22  DBA  or C57B1
mice  were given diets containing 0, 500,  1000, 1500,  2000,  3000 or 4000 ppm
copper  sulfate  equivalent  to  added concentrations of  0,  199, 398,  597, 796,
1195  or  1593  ppm  copper,  respectively.   Assuming that  mice   consume  food
equivalent  to  13%  of their  body  weight/day,  these  doses correspond  to
Intakes  of  0  (Intrinsic  copper  content not  specified  for  control  group),
25.9,  51.7,   77.6,  103.5,  155.3 and  207.1  mg  Cu/kg bw/day,  respectively.
Mice  were treated from  30 days before mating until day 19 of  gestation.
                                     -11-

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    Low doses  (500  and  1000 ppm) of copper  sulfate  stimulated embryonic de-
velopment;  Increased  Utter  size  and fetal  weight  resulted.   Higher  doses
Increased  fetal  mortality,   resulting  In  decreased  Utter   size.   Dietary
levels of  3000 and 4000 ppm  copper  sulfate caused a  low  level  of embryonic
malformation,  which was  not observed 1n  control  mice or mice  on  lower  die-
tary  concentrations.   In 55  living  fetuses  from  C57B1  mice  given  3000 ppm
copper sulfate,  1   malformed  fetus was  noted with  a defect  In  the -lumbar
vertebrae.   Among  35  living  fetuses  from  C5781  mice given 4000  ppm  copper
sulfate,   3  abnormal  fetuses  were found:  1  with  a   hernia  of  the  thoracic
wall, 1 with hydrocephalus and  1  with Mb and vertebral  fusions.  In 56 sur-
viving  fetuses from  DBA  mice   given  3000  ppm  copper  sulfate,  2  abnormal
fetuses were found, both with fusions of  adjacent  ribs.   From DBA mice given
3000  ppm  copper  sulfate, 45  fetuses  were found alive;  2  had  encephaloceles
and  2 had defects  1n their lumbar vertebrae.   The  Incidence of  terata ap-
peared to be similar  for both strains of mice tested.   The  levels of  copper
that  resulted  1n   formation  of   terata  were  considerably  higher  than  those
that  resulted  1n   toxic  effects 1n pigs  (Kline et  al., 1971); hence,  this
study 1s  not suitable  for quantitative risk assessment.
    No reports of  terata 1n  humans associated  with  oral  exposure to  copper
have been located  In the available literature.
3.3.2.   Inhalation.  No  reports of  terata  1n  humans  or  animals  resulting
from  Inhalation exposure to copper or Us  compounds  have been  located  1n the
available literature.
3.4.   TOXICANT INTERACTIONS
    Suttle and Mills  (1966a,b)  demonstrated an  Interaction  of copper  with
both  zinc and  Iron.  Groups  of  six  Large White female  pigs were  maintained
                                     -12-

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on  diets  containing  750  ppm  of  basic  copper  carbonate
H_0]  alone  or  with  500  ppm  zinc  carbonate  or  750  ppm  Iron  sulfate
(FeSO.'7H»0).   Severe   toxic   effects    (elevated  serum  copper   and   AST,
jaundice, reduced  hemoglobin,  centrllobular liver  necrosis  and  bile  canall-
cu!1 disruption) were  noted  1n control  (copper carbonate  fed)  pigs,  but the
addition  of  either   zinc carbonate or  Iron  sulfate  appeared  to  afford
protection.
    In ruminants,  an  antagonism between copper and molybdenum  has  long been
recognized  (Underwood,  1977).    High levels of molybdenum  In  feedstuff  may
protect  sheep  exposed  to high  levels  of  copper  or  may  precipitate  copper
deficiency   1n  animals   exposed   to marginal  levels   of  dietary  copper.
Excessive  dietary  levels  of  copper,  particularly when  combined with  high
levels of inorganic sulfate, have  been  shown  to elevate the  dietary require-
ment of selenium (Underwood,  1977).
                                     -13-

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                              4.   CARCINOGENICITY
4.1.   HUMAN DATA
    Pertinent  data   regarding the  cardnogenldty  of  copper  compounds  1n
humans were not located 1n the available literature.
4.2.   BIOASSAYS
    B1onet1cs  Research  Labs   (BRL,   1968)  studied  the  cardnogenldty  of
copper  hydroxyqulnonne  In   B6C3F.J  and  B6AKF,   mice.   Groups  of  18  male
and  18  female  7-day-old mice  of both  strains  were  given  1000  mg  copper
hydroxyqulnonne  In  0.5% gelatin  by  gavage until 28  days  of age, at  which
time the compound was  added to the basal  diet  (containing  5.7 ppm  copper)  at
the  rate  of 2800  ppm  (505.6 ppm copper).   Animals were  fed the  treatment
diet until 78 weeks  of  age, at which  time they were  killed.   All mice killed
or  found  dead  were  subjected to  necropsy and hlstologic examination.   Data
were compared  to  those from  positive,  negative  and  vehicle  control  groups.
No  statistically  significant  Increases   in   the   Incidence of   lymphatic
leukemias,  retlculum   cell   sarcomas,  pulmonary  adenomas   or  carcinomas,
hepatomas,  hepatic  carcinomas,  mammary   carcinomas,   skin   carcinomas   or
cancerous anglomas were observed  in orally treated mice.
    In  the  same  study,  groups  of 18  male  and  18  female  28-day-old  B6C3F,
and  B6AKF,  mice  were  maintained on  the  basal  diet  described  above and
given a single  subcutaneous  injection  of  0.5%  gelatin or  1000  mg  copper
hydroxyquinoline/kg  bw  in  0.5%  gelatin   (BRL,  1968).   The  animals,  were
observed for 78 weeks,  after  which they were  killed and subjected  to exami-
nation  as  described  previously  for  Incidence  of  tumors.    Treated  male
B6C3F.J   mice  had a significantly  (p<0.001)  Increased Incidence of  reticulum
cell  sarcomas   (6/17),  compared   with  controls  (8/141).  Male  B6AKF,  mice
evidenced no  tumor  formation.  Female  treated and  control  B6C3F,  mice had
                                     -14-

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Incidences of  1/18  and  1/154 re.tlculum cell sarcomas,  respectively.   Female
B6AKF,   treated  and  control  females  had  Incidences  of  3/18  and  5/157
retlculum  cell  sarcomas,   respectively.    Presumably,  these  Incidences  of
retlculum cell sarcomas  1n female rats were not statistically significant.
    In an earlier study, Gllman  (1962)  studied  the  carclnogenldty of cuprlc
oxide,   cuprlc sulflde  and  cuprous   sulflde  1n  2-3-month-old  Wlstar  rats.
Groups  of  30-32  rats were  given  single-dose  bilateral Injections of  20  mg
cuprlc  oxide  (16  mg copper),  cuprlc  sulflde  (13.3  mg  copper)  or  cuprous
sulflde  (16  mg copper)  Into the thighs.   Control groups  were not mentioned.
All groups  were  observed for  up to  20 months, and  survivors  were subjected
to  histopathologlcal  examination.   Surviving  to   termination  were  10/32,
19/30  and  18/30  of  the rats  treated  with cuprlc oxide,  cuprlc  sulfate and
cuprous  sulfate,  respectively.   No  Injection-site  tumors  were  observed.
Rats 1n  the  cuprlc  oxide,  cuprlc sulfate  and  cuprous sulfate groups had 0, 2
and  1   tumors,  respectively.   No further explanation  of  tumor   types  was
available.
    Haddow  and Horning  (1960)  published  bloassay results  on  various copper
compounds (Table 4-1);  no other  experimental details were provided.
4.3.    OTHER  RELEVANT DATA
    The  available  data  from  in vitro   mutagenldty bloassays   In  micro-
organisms  are not  sufficient  to allow a  conclusion  regarding  the mutagenl-
dty  of  copper.   Demerec  et  al.  (1951)  reported  positive  results  In  an
Escher1ch1a   coll  reverse  mutation   assay with  2-10 ppm copper  sulfate.
Morlya  et  al. (1983) reported a  lack  of  mutagenldty 1n  E_. coll and In Sal-
monella  typhlmurlum strains TA98, TA1535,  TA1537 and  TA1538 with up to 5 mg
copper  qulnollnolate/plate.  This compound was mutagenlc  to  S.  typhlmurlum
strain  TA100, but  only  1n  the  presence  of a  mammalian metabolic activation
                                      -15-

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                                                TABLE 4-1



                                 Tumor 1 genIdty of Some  Copper  Compounds*
Agent Under Test
Copper -dextran
L 8-Hydroxyqu1nol1ne
T copper complex
Cross-conjugated
macrocycle copper
porphyrln
Copper phthalocyanlne
Copper phthalocyanlne
tetra-3-sulfonlc acid
Copper phthalocyanlne
tetra-4-sulfonlc acid
Number and
Strain of Mice
20 stock
20 stock
20 stock
20 stock
20 stock
20 stock
Number of Weekly
Subcutaneous
Injections/Dose
6/0.1 cc of 1 In 4
dilution
39/0.1 mg
4/0.5 mg
34/0.5 mg
36/0.5 mg
25/0.5 mg
Months of
Experiment to
Date and Survivors
10(13)
10(14)
10(14)
8(17)
8(20)
8(11)
Tumors
Recorded
none
1 pleomorphlc
sarcoma
none
none
none
none
'Source:   Haddow and Horning,  1960

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system.   Up  to  5  mg of  copper  sulfate/plate failed to Induce  reverse  muta-
tions In S.  typh1mur1um strains TA98  or  TA100 either with or  without  meta-
bolic activation  (Moriya  et al., 1983).   Negative  results with  copper  sul-
fate and copper  chloride In Saccharomyces  cerevlslae  D-7 (Singh,  1983)  and
Bacillus subt111s  (N1sh1oka,  1975;  Matsul,  1980;  Kanematsu  et al.,  1980)
have also been reported.
    Results  from  several  Isolated  cell  mutagenldty  bloassays   indicate
mutagenlc potential for some copper  compounds.   Errors  In ONA synthesis have
been  Induced 1n  viruses  (Slrover  and Loeb,  1976),  and chromosomal  aber-
rations  in  rat  hepatocytes  have been  Induced  by  15-20  mM cupric chloride or
cuprlc  acetate  and 1.0  mM  copper  sulfate,  respectively.  Simian adenovirus
cell  transformation  in Syrian  hamster embryo cells  was  induced by  0.38 mM
cuprlc  sulflde  and,  to a  lesser  extent,  with 0.08 mM  copper  sulfate (Casto
et  al.,  1979).   A positive recessive  lethal  response  1n  D.  melanogaster was
observed to  result from  exposure of larvae or eggs  to copper  sulfate  (Law,
1983).
4.4.   WEIGHT OF EVIDENCE
     Data  regarding  the  carcinogenicity  of  copper  were  not  sufficient  to
enable  an   IARC  rating  on  the  carcinogenicity  of  this  element  (UrfS.  EPA,
1983a).  Cuprlc  acetoarsenlte  was  classified 1n 2A,  but this  was  based on
the  weight  of evidence  for  the arsenical moiety to  react as arsenic trlox-
ide,  and was  not related  to   the  carcinogenicity  of copper.   Applying the
criteria  for evaluating  the  overall weight of  evidence  for  carcinogenicity
to   humans  proposed  by  the  Carcinogen  Assessment  Group  of  the  U.S.  EPA
(Federal  Register, 1984),  copper  1s  most appropriately  designated  a  Group
D-Not Classified  substance.
                                     -17-

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                     5.   REGULATORY  STANDARDS  AND  CRITERIA
    Current regulatory  standards  and  criteria for copper are  shown  1n Table
5-1.
    The AC6IH  (1983) has set  the TWA-TLV for copper  fumes at  0.2  mg Cu/m3
and  the  TLV for  copper  dusts  and mists  at   1  mg  Cu/m3  of  air.   Although
Gleason (1968)  reported symptoms of  metal  fume  fever  1n workers  exposed to
0.1  mg  copper  dust/m3  air,  the  ACGIH  felt   that  extensive  Industrial
experiences with  copper  welding and  refining experience  1n  Great  Britain
supported  the  view  that  no  111  effects result  from  exposure  to  fumes  at
concentrations  up to 0.4 mg Cu/m3.
    The NAS  (1977)  has given  15  ppm copper  In  pig  feed a  GRAS categoriza-
tion.  Levels  up  to  200  ppm  are  often  used 1n  market   hogs  as  a  growth
promoter.
    The U.S. EPA (1980a), based  on  the  organoleptlc  threshold of copper, .has
set  the  ambient  water  quality  criterion  for human  effects  at  1.0  mg/a.
U.S. EPA  (1985)  recommended  this same  level  as  the criterion  for  drinking
water based on  organoleptlc criteria.
                                     -18-

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                                   TABLE  5-1
                   Current  Regulatory Standards  and  Criteria
    Criteria
     Value
  References
TLV:fumes
TLVrdust
GRAS
Ambient water quality
criterion
Dally recommended
allowance for man
0.2 mg/m3
1.0 mg/m3
15 ppm 1n pig feed
1.0 mg/9.
2-5 mg
AC6IH, 1980
ACGIH, 1980
NAS, 1977
U.S. EPA, 1980a

NAS, 1980
                                     -19-

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                             6.  RISK ASSESSHENT
    Pertinent risk  assessment  data  are  summarized In  the  Appendix of  this
report.
6.1.   ACCEPTABLE INTAKE SUBCHRONIC (AIS)
6.1.1.   Oral.   Very  few  pertinent  data  were  located  In   the   available
literature  concerning  the subchronlc  toxlclty of  orally administered  cop-
per.   Howell (1959)  studied  the   tissue  distribution  of  copper  In  stock
laboratory rats maintained for 16  months  on  diets  containing  5000  ppm copper
acetate  (250 mg copper acetate or  80  mg Cu/kg bw/day).  Both  the  liver and
kidneys  heavily accumulated  copper.   Since  only  one  treatment   level  was
used,  and presumably  no  other  criteria  of  toxlclty  were  evaluated,  this
study  was judged Inadequate  for  use In  deriving  an  MOT.  Suttle  and  Mills
(1966b)  observed elevated  serum AST activity and  jaundice 1n  pigs  fed a diet
containing  250  ppm copper  sulfate  (2.6  mg  Cu  Vkg/day).   Although  this
level  of copper 1s  slightly  less  than that  in the  Kline et  al.  (1971)  study
(3.2 mg  Cu/kg/day)  In  which  no  adverse  effects were observed.  It Is Inappro-
priate to use the dosage  from  the  Suttle and Mills (1966b)  study for deriva-
tion  of   an  AIS because the  diet  was artificially altered  to  maximize the
probability  of  copper   toxlcity.   Kline  et  al.  (1971)  exposed  groups  of 12
pigs  to   dietary  levels of  0,  150,  200 or  250  ppm  copper  sulfate for  88
days.   In an  earlier   study,  Kline et  al.   (1971) determined   that  500 ppm
dietary  copper  sulfate  depressed the rate of weight gain.  In these studies,
therefore,  250  ppm  dietary copper   sulfate appeared to be a  NOAEL.   Assuming
that  a pig  eats food  equivalent  to 5% of Its  body weight/day,  this dietary
level  corresponds  to a dally Intake of  12.5 mg copper sulfate/kg  bw, which,
assuming 5 molecules of water of  hydratlon,  corresponds to  3.2 mg  Cu/kg bw.
For  a 70  kg man,   this  exposure  would  be  equivalent to 3.2  mg Cu/kg  bw  x
                                     -20-

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70  kg *  100 =  2.2  mg  Cu/man/day.   Division  by  100  represents  an  uncer-
tainty factor  of 10,  Introduced for  Interspedes  extrapolation,  and another
uncertainty  factor  of  10 to  afford  greater protection for  unusually  sensi-
tive  populations.   This  estimate 1s  essentially  the same as  the  ADI  of 2.6
mg/day estimated by the U.S.  EPA  (1985).  It  1s  suggested  that  the  ADI  of
2.6 mg/day be  adopted here  as  the  AIS (see Section 6.2.1. for the derivation
of this number).
    Patients  suffering  from  G6PD  deficiency  may  be  at  greater   risk  from
excess levels  of copper  than  the  general population.   Excessive  copper has
been  shown   to  reduce  the  activity  of  the hexose monophosphate  shunt,  1n
which G6PD Is  apparently Involved  (D1ess  et a!.,  1970;  Boulard et al., 1975;
Calabrese et al.,  1980).   It has been  reported  that  -13%  of  the American
Black male population has G6PD  deficiency,  and  consequently  may  be at  exces-
sive  risk from the  toxic  effects  of  copper.   Individuals  occupatlonally
exposed  to   fine aerosols  of  copper  or  sprays   containing  copper  sulfate
(Plmental and  Menezes,  1975)  may be  at  additional risk,  although  1t  1s not
known what effect elevated oral Intake of copper may have  on  this population.
    The derived  AIS 1n  this  report  of  2.2 mg   for  humans  Is  consistent  with
the recommendations of the  NAS (1980) that an  "adequate  and  safe" Intake  of
2-3 mg copper  In a 70 kg man will  satisfy the nutritional  requirements and
be protective  of health.  The Food and Drug Administration  suggested  that  a
40-fold  Increase 1n the dietary  requirement  may  represent  a threshold  for
mild  to  severe  chronic  toxldty (U.S.  EPA,  1985).   It  1s  also  consistent
with  the  drinking water   standard  of  1  mg/a when  water  consumption  1s
estimated at 2 l/day (U.S.  EPA, 1980a).
    U.S.  EPA (1983c) calculated a CS  for  copper,  based  on the  elevated serum
AST activity and jaundice observed by Suttle and  Mills  (1966b)  1n  pigs fed  a
                                     -21-

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diet containing  copper  sulfate  that  contributed 2.6  mg  Cu +/kg  bw/day  for
79 days.   Since  this  was a  subchronlc  study,  the animal  MED was  divided  by
10 to  convert  to  chronic  exposure.   The result  was  multiplied by  the  cube
root of the ratio  of  the body  weight  of the pigs (33 kg)  to that of an aver-
age man to  derive  a human  MED of  0.20  mg/kg/day of Cu + or  14  mg/day for a
70 kg  human.   This MED  corresponds   to  an  RVd  of  3.8.  The elevated serum
AST activity  and jaundice  were  assigned  an RVg  of  5, since anemia  was  not
observed.   A CS of 19, the  product of RVd and RVg, was calculated.
6.1.2.   Inhalation.   No  satisfactory  reports  of   subchronlc   Inhalation
exposure of laboratory animals to copper have been  located 1n  the available
literature.  Consequently,  no subchronlc maximum dally  dose for  Inhalation
exposure 1n man can be derived.
6.2.   ACCEPTABLE  INTAKE  CHRONIC (AIC)
6.2.1.   Oral.   Howell   (1959)  exposed  rats  to  5000  ppm  dietary  copper
acetate  for 16  months.    By  histochemlcal   techniques,  both  the   liver  and
kidneys were  shown to  "heavily  accumulate"  (U.S.  EPA,  1985)  copper.  U.S.
EPA  (1985) calculated  a  dally  Intake 1n  treated  rats  of  250   mg copper
acetate/kg  bw/day,  corresponding  to -80  mg  Cu/kg bw/day.   This  figure Is -25
times  the  level  of copper  (3.2 mg Cu/kg bw/day)  to which pigs (Kline et al.,
1971)  were exposed  In  the  study from which a reasonable  subchronlc  oral
acceptable  Intake  was calculated.   Since  only  one  treatment level  1n  rats
was  studied  (Howell, 1959),  and  since  copper  was  observed  to   "heavily
accumulate" In  the Hver and  kidneys,  this study was judged unsatisfactory
for  determining an  oral  acceptable  Intake  1n  man.   Furthermore,  a dally
Intake of  -6.4  mg  Cu/kg  bw/day for 61 days  has been shown  to reduce  the rate
of  body weight  gain 1n pigs  (Kline  et  al.,  1971),  assuming that market pigs
eat the equivalent of 5% of  their body weight/day.
                                     -22-

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    Frequently,  when no suitable chronic studies are available  from which  to
derive a maximum  tolerable  oral  dose,  the maximum  tolerated  subchronlc  oral
dose 1s divided by an uncertainty factor of  10  to  derive a maximum tolerated
chronic oral  dose.   That  rationale was  considered  and rejected  for  copper,
because copper  1s an  essential  trace  element  1n  human  nutrition, and  the
body ordinarily has  homeostatlc  mechanisms to deal with  reasonably moderate
deficiencies  or   excesses.   Furthermore,   the   animal   based   subchronlc
acceptable Intake for oral  exposure,  2.2 mg/man/day,   1s  consistent  with  the
recommended dally allowance proposed  by the National  Research  Council  (NAS,
1980).  A  recommended dally  allowance  (U.S.  EPA,  1985)  of  2-5 mg/day  has
been proposed by  the National Research  Council  (NAS,  1980).   This  estimate
1s also 1n  good agreement  with U.S. EPA (1985).  In that  document  an  ADI  of
2.6 mg/day was estimated based on a human  LOAEL  for acute GI  symptoms  of  5.3
mg/day  (Chattanl  et  al.,   1965;  Semple  et a!.,  1960;  WylUe,  1957).   An
uncertainty factor  of  2  was  applied  to this LOAEL due  to a number of  con-
siderations Including the transient nature of the effects  and the essential-
ity of  copper  1n  human nutrition.  It  1s  proposed  that  the  U.S.  EPA  (1985)
ADI of 2.6 mg/day  be adopted here as the oral AIC.
6.2.2.   Inhalation.  Pertinent data regarding  chronic Inhalation of  copper
1n laboratory animals have  not been located  1n  the  available  literature.   On
the basis  of  extensive  Industrial  experience with  copper welding  and  metal
refining operations  In Great  Britain,  the  ACGIH (1983) adopted  a TLV  of  0.2
mg/m3  for   fumes  and  1.0  mg/m3  for   dusts  and mists.    Since  these  TLVs
were based  on extensive experience  with  Industrial exposure,  and  since  no
animal  toxldty  studies  were available,  1t  was  deemed  prudent  to use  the
TLVs as  a  starting  point  to derive a  maximum tolerated  chronic Inhalation
                                     -23-

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dose.  Assuming  a  man Inhales  10  m3  of air  1n  a workday and  works  5 days/
week, the dose of copper  vapor  expected  to  be Inhaled can be estimated to be
0.2  mg  Cu/m3  x  10  m3   Inhalation  rate x  5/7  days/week  =  1.4 mg  copper
vapors/day.   Similarly,   starting   with  a  TLV  of  1.0  mg  copper  dust  or
m1st/m3 of  air,  the  dose  1s  equivalent  to 7.14  mg of copper  dust  or mist/
day.  An  uncertainty  factor  of 10 1s  Introduced  to provide  an additional
safety factor for susceptible populations.  Dividing  the  values for  the dose
by  the  uncertainty  factor  of  10  results  1n  an  AIC   for  chronic Inhalation
exposure  of  0.14 mg  copper vapors  or  fumes/day and 0.71 mg  copper  mists or
dusts/day.   The  use  of   Inhalation exposure  levels   expressed  1n  units  of
mg/kg Implicitly  assumes  that  the  exposure  will  be  spread  uniformly across
the  day.
6.3.   CARCINOGENIC POTENCY (q.,*)
6.3.1.    Oral.   Blonetlc   Research  Labs  (BRL,  1968)   studied  the  carcino-
genldty  of  copper  hydroxyquinoline  1n  B6C3F1  and  B6AKF1  mice.   Dietary
levels of 2800  ppm copper hydroxyqulnollne (505.6  ppm copper,  25.3 mg Cu/kg
bw)  failed  to produce tumors  1n mice  after  77 weeks of exposure.  In another
part  of  this  study, a  single  subcutaneous  Injection  of  1000 mg  copper
hydroxyqu1no!1ne/kg   bw   was   associated  with   a   significantly   (p<0.001)
Increased incidence  of reticulum  cell  sarcomas  1n male B6C3F,  mice.  Male
B6AKF, mice manifested no  tumor  formation,  and  treated  female mice of both
strains  had no  statistically  significant  Incidence of  tumor  formation com-
pared  to  controls.    Mutagenidty  bioassays  do not clearly  delineate a car-
cinogenic role   of  copper  in  the   systems  tested.   Lack of  sufficient data
regarding carcinogenicity of copper  in humans  or  1n  animal  bioassays pre-
cluded derivations  of health advisories  on this basis.
                                     -24-

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6.3.2.   Inhalation.   The only  evidence  of human  cancer  related to  inhala-
tion  exposure  to  copper was  the suggestion  by  Plmental  and Menezes  (1975)
that  persons  exposed  to copper  sulfate  mist  In  Bordeaux mixture  (vineyard
sprayer's disease)  may be  at additional  risk for  the  development of  pul-
monary  alveolar  cell   carcinoma.   No  cardnogenlcHy  bloassays  Involving
Inhalation exposure were  found;  hence, H  was not possible to derive a  q^*
for Inhalation exposure to copper.
                                     -25-

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                                7.   REFERENCES

ACGIH  (American  Conference   of  Government   Industrial  Hyg1en1sts).   1980.
Documentation of the Threshold Limit  Values,  4th  ed.   (Includes  Supplemental
Documentation, 1981).   ACGIH,  Cincinnati,  OH.

ACGIH  (American  Conference   of  Government   Industrial  Hygienists).   1983.
Threshold Limit  Values  for Chemical  Substances and  Physical  Agents  in  the
Workroom Environment with Intended  Changes for 1984.   ACGIH,  Cincinnati,  OH.

Batsura, Y.   1969.   Electron-microscopic  study of the penetration  of  copper
aerosol  from  the lungs  into  the blood and  internal  organs.  Bryull.  Eksp.
Biol. Med.   (Rus.)  68(10): 105.   (Cited in NIOSH, 1982;  U.S. EPA,  1985)

Boulard, M.,  K.G. Blume  and  E.  Beutler.  1975.  The  effect  of copper  on  red
cell enzyme activities.  J. CUn. Invest.  51:  456-461.   (Cited  in  U.S.  EPA,
1985)

BRL  (Bionetlcs  Research  Labs).   1968.   Evaluation of Carcinogenic,  Terato-
genlc  and  Mutagenlc  Activities   of   Selected   Pesticides   and  Industrial
Chemicals.    Vol.   I.   Carcinogenic  Study   Prepared for   National   Cancer
Institute.  NCI-OCCP-CG-1973-1-1.   (Cited in U.S.  EPA,  1985)

Calabrese,   E.J.,  G.S. Moore  and  S.C.  Ho.   1980.   Low  glucose-6-phosphate
dehydrogenase  (G-6-PD)  activity in red  blood cells  and susceptibility  to
copper-induced oxldative damage.   Environ.  Res.   21:  366-72. (Cited  in  U.S.
EPA, 1985)
                                     -26-

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Callahan,  M.A.,   H.W.   SUmak,  N.W.  Gabel,  et  al.   1979.   Water-Related
Environmental  Fate  of  129  Priority  Pollutants,  Vol.  I.   Office  of  Water
Planning  and Standards,  Office  of  Water and  Waste  Management,  U.S.  EPA,
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Casto,  B.C.,  J.  Meyers  and  J.A.   DIPaolo.    1979.   Enhancement  of  vital
transformation for  evaluation  of  the carcinogenic or  mutagenlc  potential  of
Inorganic metal salts.  Cancer  Res.   39: 193.   (Cited In  U.S. EPA,  1985}

Chattanl, H.K., P.S. Gupter, S. Gallatl  and D.N.  Gupta.   1965.   Acute copper
sulphate poisoning.   Am. J.  Med.   39: 849.

Demerec,  M., G.  Bertanl  and  J.  Flint.   1951.   A  survey  of chemicals  for
mutagenlc action on E_. coll.  Am.  Natur.  85:  119.  (Cited 1n U.S.  EPA, 1985)

DICarlo,  F.J.   1980.    Syndromes  of  cardiovascular malformations  Induced  by
copper  citrate  1n hamsters.  Teratology.  21:  89-101.   (Cited  1n  U.S. EPA,
1985)

D1ess,  A.,  G.R.  Lee and G.E. Cartwrlght.  1970.   Hemolytlc  anemia  1n Wilson
disease.  Ann. Intern. Med.  73: 413.  (Cited In U.S.  EPA, 1985)

Evans,  G.W.  1973.   Copper homeostasls  1n  the mammalian  system.   Physio!.
Rev.  53: 535-570.  (Cited  In U.S. EPA, 1985)
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Federal  Register.    1984.    Environmental.  Protection   Agency.    Proposed
guidelines    for   carcinogenic   risk   assessment.    Federal   Register   49:
46294-46299.

Ferm,  V.H.  and  D.P.  Hanlon.   1974.   Toxlclty  of  copper  salts  1n  hamster
embryonic development.   B1ol.  Reprod.   11:  97-101.   (Cited 1n  U.S.  EPA,  1984)

Gllman,  J.P.W.   1962.   Metal  cardnogenesls.   II.  A  study on  the-carclno-
genlc  activity  of  cobalt, copper,  Iron  and  nickel  compounds.   Cancer  Res.
22: 158-166.   (Cited In  U.S.  EPA, 1985)

Gleason, R.P.  1968.  Exposure to copper dust.   Am. Ind.  Hyg. Assoc.  J.   29:
461-462.  (Cited In U.S. EPA,  1985)

Haddow,  A.  and  E.S.  Horning.   1960.   On  the  cardnogenldty of an  Iron  dex-
tran complex.  J. Nat.  Cancer  Inst.   24:  109-146.  (Cited  .1n U.S.  EPA,  1985)

Howell,  J.S.   1959.   H1stochem1cal  demonstration  of copper  1n  copper-fed
rats  and  In  hepatocentlcular   degeneration.    J.  Pathol.  BacteMol.    77:
473-484.  (Cited 1n U.S. EPA,  1985)

Hurley,  L.S.  and C.L.  Keen.   1979.  Teratogenlc effects  of copper.   Copper
Environ.  2:  33-56.  (Cited  1n U.S.  EPA,  1985)

Kanematsu,  N.,   M.  Hara  and  T.  Kada.   1980.   Rec assay  and  mutagenlclty
studies  on metal compounds.   Mutat.  Res.   77: 109-116.   (Cited 1n  U.S.  EPA,
1985)
                                     -28-

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KUne,  R.D.,   V.W.   Hays  and  G.L.  Cromwell.   1971.   Effects  of  copper,
molybdenum and sulfate  on performance,  hematology  and copper stores,  of  pigs
and lambs.  J. An1m.  Sd.   33:  771-779.   (CHed  1n  U.S.  EPA,  1985}

Kust,  R.N.    1979.    Copper  compounds.   JJK   Kirk-Othmer  Encyclopedia  of
Chemical Technology,   3rd ed.,  Vol.  7,  M. Grayson, Ed.   John Wiley  and  Sons,
Inc., NY.  p.  97-109.

Law,  L.W.   1983.   The effects  of  chemicals  on  the lethal mutation rates  1n
Drosophlla  melanoqaster.   Proc.   Nat.  Acad.  Sc1.   24:  546-550.    (Cited  1n
U.S. EPA, 1985)

Lecyk, M.   1980.  Toxldty  of  cupMc sulfate 1n mice embryonic  development.
Zool. Pol.  28(2): 101-105.   (CHed In  U.S.  EPA,  1985)

Matsul,  S.   1980.   Evaluation of  a Bacillus  subtillus  rec-assay for  the
detection of  mutagens which may  occur  1n  water  environments.   Water  Res.
14(11): 1613-19.   (CHed 1n  U.S.  EPA, 1985)

Morlya,  M.,  T.  Ohta, K. Watanabe,  T.  Mlyazawa,  K.  Kato  and  Y. Shlrasu.
1983.   Further mutagenlcHy  studies on  pesticides   1n  bacterial  reversion
assay systems.  Mutat. Res.   116(3-4):  185-216.   (CHed  1n  U.S.  EPA, 1985)

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Washington,  DC.
                                     -29-

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NAS  (National  Academy  of  Sciences).   1980.   Recommended Dally  Allowances,
9th ed.  Food and Nutrition  Board,  NAS,  Washington,  DC.   (Cited In U.S.  EPA,
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Plmental, J.C.  and   F.  Marques.  1969.   'Vineyard sprayer's  lung1:   A  new
occupational disease.  Thora.  24: 678-688.  (Cited 1n U.S. EPA, 1985)

Plmental, J.C.  and  A.P. Menezes.  1975.  Liver granulomas containing  copper
In vineyards sprayer's  lung.  A  new  etiology of hepatic  granulomatosls.   Am.
Rev.  Resplr. 01s.  Ill:  189-195.  (Cited in U.S.  EPA,  1985)

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Schroeder, H.A., A.P. Nason, I.H. Upton and  J.J.  Balassa.   1966.   Essential



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screening  for  potential  metal  mutagens   or  carcinogens.   Science.   194:



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pigs.   I.    Effects  of  oral  supplements   of zinc  and  iron  salts  on  the



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                                     -31-

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U.S.  EPA.    1980a.   Ambient  Water  Quality  Criteria   Document  for  Copper.
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                                     -32-

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VUlar, T.G.   1974.   Vineyard sprayer's  lung:   Clinical aspects.  Am.  Rev.



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Toxlclty In  the World Population.   Ir±: Clinical,  Biochemical  and  Nutritional



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Health.  47: 617.
                                     -33-

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                                                      APPENDIX

                                              Summary Table for Copper
CO
4k
I

Inhalation
AIS
AIC
Oral
AIS
AIC
Maximum
composite
score
Species Experimental Dose/
Exposure

NA NA
man NA

man 5.3 mg/day
man 5.3 mg/day
pig 250 ppm copper sulfate
1n diet for 79 days
(2.6 mg Cu?Vkg/day
(RVd = 3.8)
Effect

NA
NA

GI symptoms
GI symptoms
elevated serum
AST and jaundice
(RVe = 5)
Acceptable Intake Reference
(AIS or AIC)

ND NA
0.14 mg fumes/day ACGIH, 1980
0.71 mg dusts/day

2.6 mg/day U.S. EPA. 1985
2.6 mg/day U.S. EPA, 1985
19 Suttle and
Mills, 1966b
    NA = Not applicable

    ND = Not derived

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