-------
the occurrence of hyperpigmentatIon, keratosis, and skin cancer.  A study



made  in  1949-1959 indicated a higher proportion of deaths from  cancer in the



arsenical region than  in the rest of the province - 23.8% and 15.3%,.


             59
respectively.    The excess was due mainly to cancer of the respiratory and



digestive tracts in both men and women.  The excess cancer was  unrelated to



socioeconomic differences.



Experimental Animal Studies



      This section presents examples of tests that illustrate oral, topical,



and parenteral administration of arsenicals to rats, mice, and  fish.  A number



of laboratory animal studies designed to test arsenic compounds for carcino-



genicity are not included here, for such reasons as inadequate  numbers of test



animals, too short a test period, too low an exposure, and poor survival.  To



our knowledge, no adequate animal studies have been omitted that would add sub-



stantially to the examples that follow.



      In  general, animal studies have not shown carcinogenicity  for arsenic



compounds even when administered at near the maximally tolerated dosage for



long  periods.  Two notable exceptions are described first, and  then several of



the negative studies.


                    283
      In  1962, Halver    reported the occurrence of hepatomas in trout fed a



synthetic diet containing Carbarsone at 480 mg/100 g of diet.  The data were


                                 395
reviewed by Kraybill and Shimkin;    the original report is not readily avail-



able.  Of 50 trout exposed to Carbarsone, five developed hepatomas.  There



were  no  hepatomas in a large control group fed the synthetic diet without



Carbarsone.  However,  aflatoxin contamination of the diet may have been a



confounding variable.



                                         549
     More recently, Osswald and Goerttler    reported that subcutaneous injec-



tions of sodium arsenate in pregnant Swiss mice caused a considerable increase



in the incidence of leukemia in both the mothers and their offspring.  A
                                   -301-

-------
0.0057o aqueous sodium arsenate solution was injected daily during pregnancy



for a total of 20 injections, each containing arsenic at 0.5 mg/kg.  Some



groups of offspring from the arsenic-treated females were given an additional



20 subcutaneous injections of arsenic (0.5 mg/kg) at weekly intervals.  Leuk-



emia occurred in 11 of 24 mothers (46%), 7 of 34 male offspring (21%), 6 of 37



female offspring (16%), and, in the offspring given the additional 20 injections,  ,



17 or 41 males (41%) and 24 of 50 females (48%).  Leukemia developed in only



3 of 35 male (9%) and in none of 20 female offspring of untreated cortrol mice.



Furthermore, 11 of 19 mice (58%) developed lymphoma after 20 weekly intravenous



injections of 0.5 mg each of arsenic as sodium arsenate.



     Long-term studies of effects of arsanilic acid on chickens, pigs, and


                                  240
rats were reported by Frost et al.     No adverse effects were seen in the



chickens and pigs after 4 years of feeding, nor in pigs fed 0.01% arsanilic



acid for 3 generations.  Male and female weanling rats from the F» generation



of a six-generation breeding study in which 0.01% and 0.05% arsanilic acid



was fed* were held on the 0.01% arsanilic acid diet or on the control diet for



116 weeks.  The overall tumor incidence was the same in all groups, and



resembled the historical incidence of tumors in the colony, 35-45%.  The signi-



ficance of these data lies in the fact that transplacental exposure to a



carcinogen followed by lifetime exposure to the same carcinogen is often the



most sensitive technique for detecting carcinogenicity of a substance,   a



but this test was negative.



     Boutwell  a used female mice (Rockland and a specially bred strain highly



susceptible to skin tumors) in a test for cocarcinogenicity of potassium



arsenite, KAsOj.  It was tested as an initiator, both orally by stomach tube



(a total of 2.4 mg in 5 days) and locally (a total of 1.2 mg in 8 applications
                                    -302-

-------
during 5 days).  This initiating treatment was followed by topical applica-



tion of croton oil twice a week for 18 weeks.  He also tested potassium



arsenite as a promoter by daily applications (a total of 2.3 mg/week) after



a single 75->ug dose of dimethylbenzanthracene (DMBA).  The prolonged skin



applications of potassium arsenite were hyperkeratotic and ulcerogenic.  Other



experiments were done to determine whether arsenic would increase the yield of



skin cancers caused by suboptimal regimen of DMBA plus croton oil either when



given at the time of DMBA initiation or during the 24-week period of croton



oil promotion.  Under the latter condition, the mice were fed potassium arsenite



at 169 rag/kg of food.  This dietary concentration of 169 ppm (as potassium



Vtrsenite) is very high compared with the 0.5 ppm usually found  in the human



diet.   In no case was there  an effect  of arsenite on skin carcinogenesis in these



experiments.  Many tumors developed in the positive control mice beginning as



early as 6 weeks after treatment began.


                                    41
     Baroni, van Each, and Saffiotti   carried out a similar study with male



and female Swiss mice, testing the oral effects of potassium arsenite (100 ppm



in drinking water) as an initiator with croton oil promotion and as a promoter



for DMBA and urethane initiation.  Local skin applications of sodium arsenate



were tested as a promoter after initiation with DMBA or urethane.  The



arsenicals had no effect on tumorigenesis; and only a very slight degree of



keratosis was observed.


           494
     Milner    used 3 strains of mice that differed in susceptibility to the



induction of skin tumors by the application to the skin of methylcholanthrene-



impregnated paraffin, disks for 2-3 weeks.  The treated site was transplanted



syngeneically and observed for 8 weeks for tumor formation.   Arsenic trioxide



(100 ppm in drinking water) was administered either during methylcholanthrene
                                    -303-

-------
exposure, to animals with transplanted skin, or both.   Arsenic exposure pro-



duced a small increase in the yield of papillotnas in the low-susceptibility



strain, a small decrease in the high-susceptibility strain, and no effect in



the intermediate-susceptibility strain.


                 104
     Byron et al.    fed either sodium arsenite or sodium arsenate to Osborne-



Mendel rats in a 2-year study at dietary concentrations of 15-250 ppm for



arsenite and 30-400 ppm for arsenate.  No carcinogenic activity of either



material was found.  These investigators also did a 2-year arsenic feeding



experiment on dogs, with negative results; however, this was an inadequate



observation period for studying carcinogenic responses in dogs.



     Hueper and Payne     incorporated arsenic trioxide in the drinking water



(either plain or with 12% ethanol) of groups of rats and mice.  The initial



concentration of 4 mg/liter was increased by 2 mg/liter each month to a maximum



of 34 mg/liter at 15 montha.  Thus, the daily intake of arsenic trioxide ranged



from 0.1 to 0.8 mg/rat.  The administration of arsenic trioxide was continued



until 24 months.  Neither the rats nor the mice developed any cancers in sus-



pected target organs - skin, lung, and liver.



     Kanisawa and Schroeder    and Schroeder et al.    found no carcinogenic



effects on mice exposed from weaning to senescence to potassium arsenite at



5 ppm in drinking water    or on rats on the same regimen.


                 395a
     Kroes et al.     studied the carcinogenicity of lead arsenate and sodium



arsenate with SPF-Wistar-derived male and female rats.  In addition, some groups



were intubated with a subcarcinogenic dose of diethylnitrosamine to investIgate



for a possible synergistic action leading to lung tumors.  Food intake and body



weights were recorded, and complete gross and microscopic examinations were made



on all animals.  Lead arsenate that was incorporated in the diet at 1,850 ppm
                                    -304-

-------
was toxic and caused increased mortality; an adenoma of the renal cortex and



a bile duct carcinoma were found in this group but no significance can be



attached to one or two tumors in any group.  No cancer was associated with the



feeding of lead arsenate at 463 ppm or sodium arsenate at 416 ppra.  No syner-



gism with the nitrosamines was observed.  There was a high spontaneous tumor



incidence in this experiment.  The test diets were fed to female rats from the



time of parturition until the young were weaned, and these young were the test



animals.  Surviving rats were killed after 29 months of feeding.



     As Fraumeni points out, it is largely because laboratory studies have not



succeeded in producing tumors in animals that arsenic has not been accepted


                            23 2a
universally as a carcinogen.
                                    -305-

-------
EVALUATION



     Skin Cancer.  There is evidence from clinical observations and occupa-



tional and population studies that inorganic arsenic is a skin carcinogen in



man.  There is a characteristic sequence of skin effects of chronic exposure



to arsenic that involves hyperpigmentation initially, then hyperkeratosis


                                    790
(keratosis), and finally skin cancer.   This sequence has been observed under



a variety of circumstances involving chronic exposure:   potassium


                                                 530
arsenite (Fowler's solution) was used medicinally, vineyard workers used



sprays and/or dusting powders containing arsenic compounds and drank
                          86U609
                         ;, cneml
arsenic-contaminated wine, cnemlcal workers manufactured sodium arsenite for


                 572
use as a sheepdip, and residents of a southwest area of Taiwan had as their



only source of drinking water for over 45 years artesian wells contaminated by


                              714
arsenic from geologic deposits.  The similarity of responses under these



diverse circumstances is important, because studies in human populations al-



ways involve variables that cannot be controlled as in laboratory experiments;



hence, the credibility of information derived from human studies depends on the



demonstration of comparable effects under different conditions,  This require-



ment has been amply met with arsenic as a cause of skin cancer.



     The earliest skin effect of chronic arsenic exposure, hyperpigmentation



(melanosis), occurs in a dappled pattern predominantly in unexposed areas.



After the onset of melanosis, the skin begins to atrophy patchily in hyperpig-



mented areas, with the formation of keratoses which are the pathognomonic


                                   790
lesions of chronic arsenic exposure.  Only a small proportion of the keratocjs



evolve into skin cancer, and this takes place only after very many years.  The



sequence is illustrated by the Taiwan data -- the prevalence of melanosis,



keratosis, and skin cancer reaches 10% in the male, population roughly at ages


                                      714
of 18, 30, and 60 years, respectively.  Chronic exposure to inorganic arsenic





                                  -306-

-------
thus causes a slowly progressive form of patchy skin damage involving the



epidermis and adnexal structures, as well as the underlying dermis, with the



precancerous keratosea and cancers forming in the areas of chronic atrophy.



The chronic damage and tumorigenesis resulting from arsenic are similar to the



effects of ionizing and ultraviolet radiation on the skin.



     Arsenical skin cancer is readily distinguished from skin cancer induced



by sunlight, in that it occurs predominantly on surfaces that are shielded



from sunlight and multiple lesions are much more common; for example, in 428



of the 429 cases of skin cancer studied in Taiwan, there was more than one


       790
cancer.



     Substantial doses of inorganic arsenic are required to produce an appreci-



able incidence of skin cancer.  The average intake of persons treated with



Fowler's solution who developed skin cancer was around 20-30 g.  The prevalence



of skin cancer in Taiwanese men exposed to drinking water containing arsenic



at 0.3-0.6 ppm was about 15% at age 60 and over.  The normal incidence is 2-37».



On the basis of a 2-liter/day water intake for the period over which the



artesian wells were used (45 years), the total arsenic intake must have been



about 15 g, which is roughly in the same domain as that in clinical cases of



the use of Fowler's solution.  Thus, the Taiwanese data that demonstrate the



requirement for large doses of arsenic to obtain even a modest yield of skin



cancer are consistent with the relatively low frequency of skin cancer in



patients treated with Fowler's solution.  The low potency of inorganic arsenic



may explain why no skin effects have been reported in people treated for



syphilis with organic arsenicals, inasmuch as the total doses amounted to only



a few grams.  However, it is also possible that the metabolism of the organic



arsenicals is sufficiently different to preclude the occurrence of skin cancer



and other forms of arsenical damage even at higher doses.
                                   -307-

-------
     The relative frequency of melanosis, keratosis, and skin cancer was


roughly similar in the Taiwanese population and the chemical workers who


manufactured sheepdip.  On direct examination, the latter showed a 907«


prevalence of melanosis and a 30% prevalence of keratosis for a ratio of


melanosis to keratosis of 3:1.  At comparable ages, the Taiwanese showed


a ratio of about 4:1.  Two of the nine keratosis patients in the sheepdip


factory had already been treated for skin cancer, and the proportionality


between keratosis and skin cancer was about the same in Taiwan.  As in the


Taiwan experience, the sheepdi.p chemical workers had been exposed to large


doses of inorganic arsenic (up to 1 g/year), but much of this was by inhalation.


     It is possible that the trivalent and pentavalent forms of inorganic


arsenic produce the same effects on skin.  This is of interest, particularly


in view of the different metabolic patterns of trivalent and pentavalent


inorganic arsenic, the former by interaction with sulfhydryl groups and the


latter by substituting for phosphate.  The clinical use of Fowler's solution


and the manufacture of sodium arsenite as a sheepdip both involved exposure to


trivalent inorganic arsenic.  The two categories of people developed similar  skin





responses.  The Rhodesian gold miners, in whom the incidence of typical


arsenical keratoses was very high, were exposed to arsenopyrite, in which the


arsenic becomes trivalent on weathering; the reactions of arsenopyrite in the

                 *-
body are unknown.    The chemical form of arsenic in the Taiwanese artesian-well


water is also unknown; however, the reported occurrence of methane gas in the

                                                                      *•
water could preclude the existence of arsenic in the pentavalent form.


     Lung Cancer.  Of the published reports on mortality from respiratory

                                                                            AI 9
cancers in copper smelters, the most impressive is that of Lee and Fraumeni.


The study Involved a population of 8,047 white male smelter workers who were
*
 K. J. Irgolic, personal coiwnunication.
                                    308

-------
followed for 26 years; for each employee, information was available on time,

place, and duration of employment, maximal arsenic and sulfur dioxide exposures
                                                                         was  used
(descriptive, rather than numerical), and cause of death.  The life-table method/

to evaluate age-specific mortality rates for the various causes of death, and


the rates were compared with those of the states in which the smelters were.


The number of deaths available for analysis was very substantial, 1,877.   The


study demonstrated a systematic gradient for respiratory cancer according to  the


magnitude and duration of exposure to both arsenic and sulfur dioxide.  These

agents, however, were inseparably linked, because of the nature of the smelter


operations.  The amount of excess cancer was impressive, with an eightfold


increase in the workers who had the heaviest arsenic exposure for the longest


duration, i.e., more than 15 years.  The latent period — the interval between

first employment and death from respiratory cancer -- was extraordinarily long


and was inversely related to the magnitude of exposure:  34, 39, and 41 years


for the categories of heavy, medium, and light arsenic exposure.  There are


deficiencies in the study, some of which were unavoidable.  For example,  no


indication was given of whether the study population was representative of the

total worker population; the exposure rankings were based on the maximal arsenic

concentrations, rather than weighted averages derived from work histories.  No

quantitative data were available on exposure.  No attempt was made to validate

the stated causes of death.  No smoking histories were obtained.  However, none

of these deficiencies could be seriously regarded as invalidating the conclusions


of the study.

     The Kuratsune report dealt  with a smaller study that compared lung-cancer


mortality rates calculated from the 22 deaths that occurred in a 30-year period


in a smelter town with the lung-cancer experience in the same period in a

                                        398
neighboring city and in Japan as a whole.  The standardized mortality rate for
                                    -309-

-------
males in the smelter towns was four times higher than that for the rest of



the country, but equal to that for women.  This fourfold excess is comparable



with the 3.3-fold excess observed in the Lee-Fraumeni study.  Although many of



the men in the town worked in the refinery, a much higher proportion of the

                                         heavily exposed to arsenic as

lung-cancer cases, compared with controls, occurred in men who were/smelter



operators.  As in the case of the Lee-Fraumeni study, the latent period from



first exposure to the diagnosis of lung cancer was very long, ranging from



26 to 48 years.  The duration of employment was also very long, with a median



of about 30 years, although two cases occurred in people who worked for only



2-3 years.



     Two lung-cancer studies of the American Smelting and Refining Company



smelter have produced conflicting results.  The 1963 Pinto and Bennett report



examined the proportional mortality from lung cancer in a total of 229 deaths

                        coo

in the period 1946-1960.     This study dealt only with pensioners and workers



who died during their employment and did not include people who had left the



plant.  The reported data showed that the 18 lung-cancer deaths in the plant



population as a whole indicated a rate that was higher than the rate in the state



of Washington.  However, the excess lung cancer for the plant as a whole was due



to the high occurrence in controls, i.e., in workers who were considered not to

                           and Strong,

have arsenic exposure.  Milham /by contrast, found, In the years 1950-1971, that



there were records of 39 deaths due to respiratory cancer in Pierce County (the


                                                                        491
smelter locale) in people who were stated to have worked at the smelter.



Application of U.S. mortality rates to the published figures for the smelter



population at risk yielded an expected number of 18 respiratory-cancer deaths,



compared with the 39 deaths observed.


                *                                                                582
    Pinto et al.  recently resolved the discrepancy between the Pinto and Bennett


                     491
and Milham and Strong    papers in a study of the same smelter that reevaluated


                                                           582
the exposure categories used in the Pinto and Bennett paper    (which were



apparently in error) and also included a longer observation period and therefore more
 Pinto, S. S. ,  V. Henderson, and P. Enterline.   Mortality experience of arsenic:

 exposed workers.  Unpublished data.

                                      -310-

-------
deaths.  The data, shown in Table 6-2, include a total of 32-respiratory-cancer
cases and show a progressive increase in standardized mortality ratio with
increasing arsenic exposure.  The arsenic-exposure index was calculated as a
weighted average based on urinary arsenic concentration and duration of
employment.  It is of interest that the eightfold excess in respiratory cancer
for workers with the highest exposures and the threefold excess for all the
smelter workers reported by Pinto ejt al.   were very close to the figures re-
                          412               398
ported by Lee and Fraumeni    and Kuratsune.
     The studies described here  indicate that excess respiratory cancer occurs
in copper-smelter workers as a function of the magnitude and duration of
exposure to arsenic.          with latent periods of three to four decades from
the  time of initial exposure.  However, the studies do not permit a resolution
of the issue of whether concomitant exposure to sulfur dioxide and other smelter
dusts  is necessary for the carcinogenic response.  Evidence from studies
involving  entirely different circumstances of exposure -- including workers in
three pesticide manufacturing plants,317'550'Baetjer — *!•** vintners who
applied pesticides,   '    and Rhodesian gold miners   a -- suggests that sulfur
dioxide and other unspecified smelter dusts are not essential cofactors for
the  respiratory carcinogenicity  of arsenic.  All the nonsmelter studies have
obvious limitations,  but the lung-cancer excess in each study was relatively
large and, taken as a group, they provide significant evidence that arsenic is
a lung carcinogen.
     The Hill-Faning  study of 75 deaths in a sheepdip factory used the indirect
method of  proportional mortality to evaluate the small group of 22 deaths from
cancer; seven of them were cancers of the respiratory tract, compared with an
expected 2.4 deaths.     The Dow arsenic workers    were evaluated in two ways:
first by an analysis  comparing death records in terms of the 16.2% proportional
mortality  from lung cancer (28 deaths) in 173 chemical-worker deaths, compared
with 5.77,  for 1,809 control-case deaths; and then as a retrospective cohort
 Pinto, S. S., V. Henderson, and P. Enterline.  Mortality experience of arsenic
 exposed workers.  Unpublished data.  (See Table  6-5)
  Baetjer, A., M. Levin, and A. Lilienfeld.  Analysis of mortality experience
  of Allied Chemical plant.  Unpublished data.
                                   -311-

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-------
study the mortality from respiratory cancer (obtained from the records used

in the first approach) among 603 persons Identified as having worked in the

arsenic plant from 1940 to 1973 was compared with the mortality for the

corresponding U.S. white male population.  The two approaches gave essentially

the same results, namely a threefold to fourfold excess.  However, the

puzzling aspect of the data is that almost 60% of the respiratory-cancer deaths

were in people who had worked with arsenic for less than a year, three decades

earlier.  Most of the arsenic workers were unskilled short-term employees, of

whom a large proportion left the company after a brief period of employment.

The follow-up study, however, dealt only with the people who remained in the

company.  A confirmation of the excess lung cancer in a follow-up of short-term

arsenic workers who left the company would be very useful.  Nevertheless, there

were about a dozen cases in people who worked longer than a year and who were

in the highest dose categories, where the excess risk was maximal, fourfold to

sixfold.  It is possible that the apparent twofold excess in lung cancer in the

lower exposure categories, including those who worked with arsenic for less

than a year, would not be ascribable to arsenic, because there was no change in

cancer risk over a very wide range of total doses (42-1556 mg).  Furthermore,

these low dose categories consisted predominantly of short-term unskilled

workers who as a group might have had higher exposures to other hazardous

chemicals than the controls.

     The Allied Chemical Company pesticide manufacturing operations produced

a range of products, including some arsenical compounds.   A preliminary study

of the proportional mortality among retired employees showed a sevenfold excess

of lung cancer that accounted for about 40% of all deaths (A. M. Baetjer, personal
communication).
Both the Dow and Allied studies also showed a few excess deaths from lymphoma

and Hodgkin's disease.  The results of a more detailed study of the Allied

Chemical Company that is now in progress will be very useful.


                                 -313-

-------
     Arsenic sprays and dusts were widely used in Germany between 1925 and




1942, at which time they were banned.  '      Vineyard workers also drank wine




containing arsenic.  Hundreds of workers  developed acute and chronic arsenic




poisoning.  In the 1950's, vineyard workers with lung cancer began to appear




in hospitals serving the vineyard regions.  An association between arsenic and




lung cancer is further suggested by the high proportion of vineyard workers




with lung cancer who had the characteristic hyperpigmentation and keratoses




associated with chronic arsenic exposure.




     The same high degree of association of skin arsenism and lung cancer




occurred in Rhodesian gold miners who were heavily exposed to arsenopyrite




dust.      In the period 1957-1963, the occurrence of 37 cases of lung cancer




in gold miners represented an incidence of 206/100,000, compared with 34/100,000




for adult males in the Gwanda region of Rhodesia.  This represents a sixfold




difference in lung cancer in miners.




     The probability of death from lung cancer in persons with keratosis shown




in Table 6-6 ranges from 32 to 56%, which is roughly 5-10 times higher than




might be expected.
                                   -314-

-------
                                Table 6-6


      The Frequency of Lung Cancer In Persons with Keratoses Who Had


                    Heavy Exposure to Arsenical Dusts
(a)
Cases of
Keratosis
40
16
30
12
(b)
Cases of
Lung Cancer
13
9
10
5
b/a
7.
32
56
33
42
Refer'
ences
54 8a
86
609
572
     Rhodesian gold miners


     Vintners (Braun)


     Vintners (Roth)


     Sheepdip workers
               Total             98             37             38


      Assumes that 41 chemical workers who died in 1910-1943 had the
      same skin changes as chemical workers examined in 1946.
     The data suggest that there is a very high risk of lung cancer when


 the exposure to  inorganic arsenic dust  is high enough to cause keratoses.
     Liver Cancer.  The only evidence that arsenic is a. liver carcinogen comes


from German vintners.  Thirteen of the 27 persons whose autopsies were reported

      609
by Roth had cirrhosis, and three had angiosarcoma, a rare form of liver cancer


associated with exposure to a vinyl chemical and Thorotrast.  Only two cases of
angiosarcoma have been reported in people treated with Fowler's solution.


There is no evidence of either cirrhosis or        liver damage in any of the


other studies on arsenic.  It is possible that the combined effect of a high


alcohol intake and arsenic is responsible for the unusual forms of cirrhosis and


liver cancer observed in vintners.  It should also be pointed out that the


chemical form of arsenic in wine is unknown.


                                   -315-

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     Experimentally Induced Cancer.  The fact that there is no established
method for producing cancer by treatment with any form of arsenic in an animal
model system remains an enigma.  One must conclude either that arsenic is not
a carcinogen or that particular circumstances not yet achieved are essential
to demonstrate a role for arsenic in experimental carcinogenesis.  A conclusion
that carcinogenesis by arsenic is restricted to humans (or cows, horses, and
    1 ft 1 a
deer    ) is highly suspect.  Therefore, much effort should be spent In attempt-
ing to find conditions in which the presence or absence of arsenic determines
the appearance or nonappearance of cancer in an animal model.  Some questions
need to be explored (their answers may account for the variable incidence of
human cancer associated with arsenic exposure):
•    Potassium arsenite, arsenic trioxlde, and possibly other compounds of
arsenic appear to have an unusual propensity to alter epithelial morphology
(at least in humans), often acting as irritants and causing hyperplasia, as
well as hyperkeratosis.  Thus, appropriate forms of arsenic should be tested
with known lung carcinogens for synergistic action.  Possibilities include the
ferric oxide-benzopyrene model in the hamster developed by Saffiotti et al.
and the sulfur dioxide-benzopyrene inhalation model of Kuschner and Laskin.
Controls designed to test exposure to arsenicals alone should be included;
proper and long-term inhalation studies have not been done.
                                41             77a
     Although both Barone et ajU   and Boutwell    based their tests in mouse
skin on a possible cocarcinogenic role for arsenic and found none, additional
experiments of this nature are reasonable.
     Moreover, because morphologic changes in epithelial tissue are ascribed to
arsenic, and because vitamin A and some retinoids control normal epithelial
           674a
morphology,     it is appropriate to design experiments in which vitamin A
deficiency is induced in animals as a test system for arsenical carcinogenicity
(and cocarcinogenicity).  In experimental animals, vitamin A deficiency
increases susceptibility to chemical carcinogenesis, and high
                                  -316-

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dietary concentrations of retinoids have remarkable ability to prevent chemical




carcinogenesis in epithelial tissues,     including skin, breast, and lung.




     Again, because arsenicals alter epithelial morphology, the possibility that




the function of mucus-secreting cells or of the ciliated cells of the lung is


                                                       S76
interfered with by respirable particles bearing arsenic    should be investigated.



Interference with mucus secretion or ciliary action would facilitate the action




of a carcinogen entering the lungs, such as tobacco smoke.




•    Because compounds of arsenic and the heavy metals that may be associated with



them are enzyme poisons, it is possible that chronic exposure to abnormal amounts




of these substances partially  poisons enzymes that inactivate carcinogens.



Model systems might be devised to test this possibility.




•    The interaction of arsenic with some essential nutrients, such as sodium




selenite and potassium iodide, is known.  This should be considered in designing




animal models.

              ,                      arsenite and sodium arsenate are

     Peoples   a has shown that potassium /         detoxified via methylation




pathways.  Biologic changes attributable to arsenic might be accentuated in



animals fed diets that are low in labile methyl groups.




•    The administration of some carcinogens to pregnant females may result in




an unusually high Incidence or early development of cancer in the offspring.


                                                549
Because one such test, by Osswald and Goerttler,    resulted in an unusual



incidence of leukemia in mice, it is especially urgent to design appropriate




transplacental tests.  Repetition of such a test is essential.  The credibility



of the Osswald and Goerttler study is limited by their failure to give the



vehicle solution to the controls.




•    Because of the failure of repeated tests in lower animals to show carcino-




genicity due to arsenicals, consideration should be given to the use of nonhuman



primates as test animals.




     These are only a few examples of approaches to the problem of ascertaining




whether an animal model may be devised to account for the association of human



cancer with exposure to arsenic.


                                    -317-

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                                 CHAPTER 7




                          SUMMARY AND CONCLUSIONS
CHEMISTRY
     The compound of arsenic produced in largest quantity is arsenic trioxide,




As 0~.  It is a byproduct of the copper smelting industry.  Arsenic exhibits




oxidation states of III and V and forms a great variety of inorganic and




organic compounds.  In addition to arsenic trioxide, some widely encountered




inorganic compounds are arsenic pentoxide, As^O-; arsenous acid, H^AsOoj




arsenic acid, H~AsO, ; tetraarsenic tetrasulfide (realgar), As,S, ; arsenic tri-




sulfide (orpiment), As.S,; and arsenic pentasulfide, As^S,-.  Some of the more




common organic compounds are methylarsonic acid, CILAs(O)(OH)„; dimethylarsinic




acid, (CH3)2As(0)(OH); methyldihydroxyarsine, CH As(OH>2; dimethylhydroxyarsine,




(CH3)2AsOH; trimethylarsine, (CHO-jAs; and trimethylarsine oxide, (CH3)3AsO.




Some aromatic arsenic derivatives with veterinary and medicinal uses are




4-aminophenylarsonic acid, 3-nit:ro-4-hydroxylphenylarsonic acid, 4-nitro-




phenylarsonic acid, and 3~nitro-4-ureidophenylarsonic acid.




     Cationic species of As(III) are probably not present in aqueous solution.




Arsenous acid likely exists as As(OH)3>  The fact that the hydroxides of iron(II),




iron(III), chromium(III), and aluminum strongly adsorb or form insoluble pre-




cipitates with arsenites and araenates is important in the control of arsenic




pollution.  The ability of various molds and bacteria to convert arsenic com-




pounds to various methylated arsines is well known.  Because the methylated




arsines are sparingly soluble in water, volatile, and sensitive to air, they are




returned to the environment as methylarsonates, dimethylarsinates, and tri-




methylarsine oxide.  Arsenic-sulfur bonds are less subject to hydrolysis than




arsenic-oxygen bonds, and the formation of arsenic-sulfur bonds with sulfur-




containing biologic molecules is considered to be of great importance.





                                    -318-

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DISTRIBUTION




     Arsenic is ubiquitous in the environment and is found in all living




organisms.  Natural sources include a variety of sulfur-containing minerals




of which arsenopyrite is the most common.  The amounts of arsenic in soil and




water depend largely on the geologic inputs from mineral weathering processes,




whereas the amounts in indigenous plants and animals reflect species differ-




ences.  Some species of marine plants, such as algae and seaweed, and marine




organisms, such as crustaceans and some fish, often contain naturally high con-




centrations of arsenic.




     Manmade sources of arsenic are generally byproducts of the smelting of




nonferrous metal ores, primarily copper and to a lesser degree lead, zinc, and




gold.  In the United States, the sole producer and refiner of arsenic trioxide




is the copper smelter of the American Smelting and Refining Company in Tacoma,




Washington.  Major imports of arsenic come from Sweden, the world's leading




producer.
                                   -319-

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     The largest use of arsenic is in the production of agricultural pesticides,




under the categories of herbicides, insecticides,  desiccants,  wood preserva-




tives, and feed additives.   Arsenic trioxide was the raw material for the older




inorganic pesticides, including lead arsenate,  calcium arsenate>  and sodium




arsenite.  The newer major  organic arsenical pesticides include two herbicides,




monosodium and disodium methanearsonate and cacodylic acid,  and four feed




additives in current use are




substituted phenylarsonic acids.  Arsenic has several minor  uses, primarily as




an additive in metallurgic  applications, in glass  production,  as a catalyst in




several manufacturing processes, and in medicine.   Arsenical drugs are still




used in treating tropical diseases, such as African sleeping sickness and




amoebic dysentery, and are  used in veterinary medicine to treat parasitic dis-




eases, such as heartworm (filariasis) in dogs and  blackhead  in turkeys and




chickens.




     The major arsenic residues resulting from use of agricultural pesticides




and fertilizers are found in soils and to a lesser degree in plants and animals




living on contaminated soils.  The highest pesticide residues occur primarily




in orchard soils that received large applications  of lead arsenate.  Large




accumulations of arsenic also occur in soils around smelters.   Two important




closely related effects measurable in plants are arsenic residues and phyto-




toxicity.  Some soils that  received massive applications of  arsenate are cur-




rently incapable of supporting plant growth.




     Arsenic in air has three major sources:  smelting of metals, burning of




coal, and use of arsenical  pesticides.  Two known acute incidents of arsenic




pollution from smelters have occurred in the United States.   The most serious
                                    -320-

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 air pollution problem, however, is associated with manufacturing processes and




 occupational hazards to workers.  Some arsenic in water results from




 industrial discharges.  Several endemic poisonings of water supplies have been




 reported.




      Safe disposal of arsenic wastes still constitutes a major administrative




 and technologic problem.  The major sources of arsenical wastes are residues




 in empty pesticide containers; surplus pesticides stored by government agencies,




 manufacturers, state and municipal facilities, and users; and soil contaminated




 by extensive use of arsenical pesticides.  Recommended procedures for manage-




 ment of arsenical wastes are recycling and reuse (preferred), long-term storage,




recovery of other metals and long-term storage of arsenic trioxide, and disposal




 in landfill sites.




      Several arsenic cycles have been proposed to interrelate the source, emis-




 sion, movement, distribution, and sinks of various forms in the environment.




 Arsenic is continuously cycling in the environment, owing to oxidation, reduc-




 tion, and methylation reactions.  Man's activities can alter the distribution




 of arsenic in finite geographic areas or in selected components of the environ-




 ment, but man has little control over the natural processes.




 METABOLISM




      Arsenic compounds must be in a mobile form in the soil solution in order




 to be absorbed by plants.   Except for locations around smelters or where the




 natural arsenic content is high, the arsenic taken up is distributed  throughout




 the plant body in less than toxic amounts.




      In nature, arsenic absorption by plants from the air is negligible.  Al-




 though smelter fumes and dusts may deposit on plant leaves, there is no evidence




 that arsenic from this source is taken into plants.
                                     -321-

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     Translocation of arsenicals in plants is demonstrated by the fact that



arsenical solutions applied to foliage of some weeds results in the killing




of root tissue.  Metabolic experiments with radiolabeled organic arsenic compounds




indicate that these compounds or metabolites thereof form complexes with some




plant constituents.




     Bacteria and fungi can metabolize inorganic arsenic salts to form




methylated derivatives.  Algae can biosynthesize complex organic arsenicals




that are associated with the lipid fraction of these microorganisms.  Mollusks




and crustaceans can contain rather high concentrations of arsenic, but there




appears to be no relationship between their arsenic content and the collection




date or geographic location; this suggests that industrial pollution is not a




factor.  Fish also can contain arsenic, which apparently is derived from their




diet.  The arstnic that occurs naturally in seafood is metabolized quite




differently from inorganic arsenic.  The form of arsenic in shrimp, for example,




is not retained by the human body and is rapidly excreted.




     The rat has a unique arsenic metabolism that renders it unsuitable for




metabolic studies with arsenic compounds.  This rodent stores arsenic in the




hemoglobin of its red cells, which release the arsenic only when they break




down.  The resulting very slow excretion led to the belief that arsenic is a




cumulative poison.  Trivalent sodium arsenite seems to be almost entirely




oxidized to pentavalent sodium arsenate in vivo.  Evidence of the opposite




process - i.e., the in vivo reduction of arsenate to arsenite - is much less clear.




     Arsenic in normal urine of man, dog, and cow is principally in




the methylated form.  When the dog and cow are fed large doses of trivalent or




pentavalent inorganic arsenic, about half the arsenic appears in the urine as




methylated derivatives.  This methylation process is true detoxicification,
                                    -322-

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   inasmuch as methylarsonates and dimethylarsinates are only one two-




   hundredth as toxic as sodium arsenite.




   EFFECTS ON ANIMALS AND PLANTS




        A number of different factors can influence the toxicity of arsenicals,




   including chemical form, physical form, the mode of administration, species,




   and criterion of toxicity.  There are several reports in the literature that




suggest that arsenic can exert biologic effects at concentrations below those




   generally considered "safe," but the physiologic significance of such findings




   is not known.




        The trivalent forms of arsenic apparently exert their toxic effect chiefly




   by reacting with the sulfhydryl groups of vital cellular enzymes.  Pyruvate




   dehydrogenase seems to be a particularly vulnerable site in metabolism, because




   it contains the dithiol lipoic acid that is especially reactive with trivalent




   arsenicals.  The biochemical basis for the toxic action of pentavalent arsenic




   compounds is known with less certainty, but such arsenicals may well compete




   with phosphate in phosphorylation reactions to form unstable arsenyl esters that




   spontaneously hydrolyze and thereby short-circuit energy-yielding bioenergetic




   processes.




        The use of phenylarsonic animal feed additives as recommended is beneficial




   and does not constitute a human or animal health hazard.  Animal losses and




   excessive arsenical residues in poultry and pork tissues occur only when the




   arsenicals are fed at excessive dosages for long periods.  The mechanism of




   action of phenylarsonic animal feed additives remains obscure, and these com-




   pounds are for the most part absorbed and excreted without metabolic change.
                                      -323-

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     Toxicoses caused by the phenylarsonates are manifested by an entirely



different syndrome from those caused by the inorganic and aliphatic organic




arsenicals.  The latter produce the typical signs and lesions usually asso-




ciated with arsenic poisoning, whereas the former are less toxic and produce




     demyelination and gliosis of peripheral and cranial nerves.




     Poisoning of forage-eating livestock by inorganic and aliphatic organic




arsenical compounds, especially those used as herbicides and defoliants, has




been reported.  Most cases result from accidental or careless contamination




of forage that becomes accessible to livestock.




     The large-scale use of arsenicals in the United States has caused some




scientists to suspect that the use of these compounds may have a deleterious




effect on wildlife.  However, there is little evidence to confirm such suspi-




cions in the scientific literature.  Wildlife kills that have been attributed




to arsenic compounds were all associated with misuse of the compounds in




question.  But several laboratory studies have shown that wild species are




generally more sensitive to arsenic poisoning than many domestic species;




therefore, some ecologic vigilance is appropriate.




     Most data on the effects of arsenicals on aquatic organisms, particularly




on fresh-water organisms, were collected in short-term, direct-lethality




studies.  Practically nothing is known about the sublethal long-term effects




of arsenic, singly or in combination with other pollutants, on aquatic




organisms.




     Although early workers were not able to demonstrate any adaptation in




animals to toxic concentrations of inorganic arsenic, some recent work suggests




that there may be a rather limited adaptive response to inorganic arsenicals




under some conditions.
                                   -324-

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     Abnormal physiologic responses have been noted in animals exposed to




arsenic trioxide aerosols at concentrations considerably below currently




accepted air quality standards.  Unfortunately, these experiments were carried




out with the rat, which has a unique ability to accumulate arsenic and is




therefore a poor animal model for studying arsenic metabolism.  It is difficult




to draw valid conclusions about the public health or environmental implications




of these investigations.




     High concentrations of arsenicals have been shown to decrease the ability




of mice to resist viral infection, presumably by inhibiting interferon forma-




tion or action.  However, low concentrations of arsenicals appear to enhance




the antiviral activity of interferon.




     Arsenic is known to protect partially against the effects of selenium




poisoning over a wide variety of conditions.  Arsenic decreases the toxicity




of selenium by enhancing its biliary excretion, thus clearing it from the liver,




the primary target organ in selenosis.




     Preliminary results have suggested a role for arsenic as a nutritionally




essential trace element.  Improved methodology in trace-element research - such




as the use of ultrapure water, highly refined diets, and plastic animal hous-




ing - apparently have enabled nutritionists to uncover a function for arsenic




in normal metabolism.




     The biologic effects of arsenic compounds on microorganisms appear to be




mediated very much by the same mechanisms as in mammals.  However, some micro-




organisms have a substantial ability to adapt to toxic concentrations of




arsenicals.  This adaptation seems in most cases to be due to decreased perme-




ability of the microorganism to arsenic.
                                  -325-

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     Arsenicals clearly can be toxic to plants, but the biochemical basis




for such toxicity is less understood than that of the toxicity of arsenicals




to animals.  As in animals, arsenates are generally less toxic to plants than




arsenites.  One of the first symptoms of plant injury by sodium arsenite ia




wilting caused by loss of turgor, whereas the symptoms due to arsenate do not




involve rapid loss of turgor, at least through the early expression of




toxicity.




     The phytotoxicity of organic arsenical herbicides is characterized by a




relatively slow kill; the first symptoms are usually chlorosis, cessation of




growth, and gradual browning followed by dehydration and death.  Several




variables can influence the response, including stage of growth, senescence,




moisture availability, temperature, light intensity, and insect or mechanical




wounding of foliage before treatment.




     Arsenic can interact with several plant nutrients in either soils or




nutrient solutions.  Phosphate can increase or decrease the toxicity of arsen-




icals, depending on the experimental conditions.  The toxicity in some species




grown on arsenic-contaminated soils could be reduced by foliar or soil applica-




tion of zinc or iron.




EFFECTS ON MAN




     The past medicinal use of: inorganic arsenic preparations has provided the




basis for reasonably clear definition of the consequences of chronic systemic




arsenic exposure - specifically, characteristic hyperkeratosis and, less fre-




quently, irregularities in pigmentation, especially on the trunk.  Association




of these features with other, less common disorders, such as arterial insuffi-




ciency and cancer, in exposed populations must be regarded as supportive




evidence of a causal function of arsenic.  It should also be noted that many
                                   -326-

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studies of populations "at risk" have failed to evaluate cutaneous changes


adequately.  Proper examination of the skin of people subjected to chronic


low-dose arsenic exposure has the potential for providing valuable information


related to the dose and duration of exposure necessary to cause changes in


given populations.  In a word, these benign skin lesions may be regarded as


sensitive indexes of exposure to an agent that has potentially serious con-


sequences.


     The present generation of physicians has not used arsenic and has little


direct knowledge of its toxic manifestations.  Thus, the "index of suspicion"


of the average practitioner may be relatively ineffective in diagnosing


isolated cases of arsenic toxicity.


     There is also considerable reason to believe that, judiciously used,


arsenic may have therapeutic value.  The time may be ripe to rediscover an old


remedy with modern analytic techniques.


     Several occupational and nonoccupational episodes of arsenic toxicity


have occurred.  Two of the best characterized and yet least known nonoccupa-


tional episodes occurred in Japan in 1955.  One involved tainted powdered milk;


the other, contaminated soy sauce.  In the former, 12,131 cases of infant


poisoning were recorded, with 130 deaths.  Evidence of severe damage to health,


including retarded growth and brain dysfunction, was found in a follow-up study


15 years later.


     Experimental teratogenic effects of arsenic compounds have been reported,


but none of the studies has  been sufficiently exhaustive to allow accurate


assessment of the human hazard.  For example, the doses that were administered


to achieve effects far exceeded likely environmental exposure, and accurate

                                          »
no-effect doses were generally not determined.
                                    -327-

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     There is some evidence that arsenicals can be mutagenic in humans:  an



increased incidence of chromosomal aberrations was observed in phyto-




hemagglutinin-stimulated lymphocyte cultures prepared from psoriasis patients




who had been previously treated with arsenic.




     There is strong epidemiologic evidence that inorganic arsenic is a skin




and lung carcinogen in man.  Skin cancer has occurred in association with




exposure to inorganic arsenic compounds in a variety of populations, including




patients treated with Fowler's solution, Taiwanese exposed to arsenic in




artesian well water, workers engaged in the manufacture of pesticides, and




vintners using arsenic as & pesticide.  The Taiwan data demonstrated a gradient




of incidence with degree of exposure and age.  All these populations had a




pathognomonic sequence of skin changes leading to cancer.




     Lung cancer has been observed to be associated with inhalation exposure




to arsenic in copper smelters, workers in pesticide manufacturing plants,




Moselle vintners, and Rhodesian gold miners.  Two of the three smelter studies




showed a gradient in the incidence of lung cancer with the degree of arsenic




exposure; one of these studies also suggested that sulfur dioxide may be a




carcinogenic cofactor for the lung.




     Although hemangioendothelioma has been reported occasionally in people




who have been exposed to arsenic, the case for arsenic as a liver carcinogen




is not clear.
                                   =328=

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The absence of a useful animal model is a serious handicap to the study of




arsenic as a skin carcinogen and is probably due to metabolic differences be-




tween humans and the animals tested so far.  The failure to induce skin cancer




in test animals is perhaps not surprising, inasmuch as neither melanosis nor




keratosis has been duplicated in animals and these effects appear to be




inseparably linked to the tumorigenic action of arsenic in the skin of man.




The carcinogenicity of arsenic for the lung in animals has not yet been




evaluated by inhalation studies.




MEASUREMENT




     The  preparation of material  for the  determination of arsenic requires the




usual  care to  ensure that the portion  of  the sample submitted for analysis




truly  represents  the whole.   Special hazards related  to arsenic  compounds are




the  possible loss  of arsenous oxide by volatilization and the rapid adsorption




of some arsenic compounds from  solution onto the walls of storage vessels.




     If total  arsenic  is to  be  measured in plant or animal tissue or  in  coal,




the  sample is  first wet-ashed with  some combination of nitric, perchloric, and




sulfuric  acids.   Arsenic originally present in the sample at very low concentra-




tions  must often  be preconeentrated before it can be  measured.   If  the sample




is a solution, the arsenic can  be coprecipitated on metallic hydroxides  or




precipitated with  organic reagents.  It can also be isolated from its original




matrix by liquid-liquid extraction  or  by  volatilization as a trihalide or as




arsine.
                                 -329-

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      Until  recently,  total  arsenic was usually determined colorimetrically,

 by either  the molybdenum blue  method  or  the  silver diethyldithiocarbamate

 method.  Arsenic  is now  usually determined by atomic absorption, with  the

 sample  solution introduced  into a flame  as an aerosol or deposited as  a

 droplet  inside a  tube or on a  metallic strip, which is then  strongly heated.

 Greater  sensitivity has  been achieved with atomic absorption, however, by

 converting  the arsenic to arsine and  introducing this gas into  a heated  tube.

 Equal sensitivity can  be  achieved by  introducing the arsine  into an arc  in

 helium and measuring the  resulting spectral emission.   Low detection limits

 for arsenic can also be  reached by neutron-activation analysis  (often without

 chemical treatment).   Electrochemical methods, such as differential pulse

 polarography,  can achieve comparable  sensitivity in the presence of natural

 pollutants  (e.g., sludge).

 CONCLUSIONS

     Environmental contamination with and human exposure to arsenic compounds

have resulted  from incidents of air pollution from smelters,  the improper use

 of arsenical pesticides,  and episodes of tainted food  and drink.  The degree of

 arsenic air pollution due to amelter operations and  pesticide use should decrease
                            and environmental
 if currently proposed occupational/standards  are promulgated.  The technical and

 economic feasibility of the changes in engineering controls or work practices

needed to achieve compliance with such standards,  however,  has yet to be

determined.

     Although the total amount:  of arsenic injected into  the atmosphere in the

United States as a result of coal-burning is  very large,  the  sources  of such air

pollution are widely dispersed, and  arsenic  exposure due  to  fossil-fuel com-

bustion does not seem to  constitute  a health  hazard.  This  contrasts  with the

situation in some  other countries (e.g.,  Czechoslovakia), where the arsenic
                                 -330-

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content of coal is high and high ambient air concentrations of arsenic result

Although petroleum generally contains only small quantities of arsenic, oil

from shale can contain significant amounts; therefore, if use of this fo.<;nii

fuel becomes common, removal of arsenic from the oil and/or more careful

environmental monitoring of arsenic is indicated.

     The food supply normally contains small amounts of arsenic, but these are

not considered harmful.  Some seafood naturally has appreciable concentrations

of arsenic, but in such a form that it is rapidly and completely excreted by

humans after ingestion.  Arsenic residues in foodstuffs due to arsenical pesti-

cide or feed additive use do not seem to warrant concern.  There have been

isolated epidemics of food poisoning due to arsenic as a result of manufactur-

ing accidents, but they are rare.

     Water supplies generally contain negligible quantities of arsenic, although

some cases of endemically poisoned waters have been reported.  Industrial

effluents have been shown to contain arsenic, but the self-purifying tendency

of rivers and streams and improved quality of waste-water discharges should
help to minimize this problem.
     The use of arsenical pesticides in food crops declined greatly after intro-

duction of the chlorinated hydrocarbon and organophosphorus chemicals.  However,

as more and more restrictions are placed on the use of the latter two families

of compounds, the use of arsenical pesticides may once again assume importance.

If this occurs, more careful monitoring of arsenic in the environment and food

supply would be imperative.

     Our greatest area of ignorance about arsenicals in the environment has to

do with the ecologic cycling of arsenic compounds.  Little or no quantitative

information is available regarding the fate of arsenicals in the ecosphere, so
                                  -331-

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 it is not possible to state with certainty whether arsenic is building up in




 any sector of the ecosystem.  For example, organic arsenicals are widely used




 as herbicides and desiccants, but we do not know whether such use will eventu-




 ally render the soil phytotoxic, as has happened in some orchards in which lead




 arsenate was heavily applied.  More research is needed to investigate such




 problems.




      Suitable methods for arsenic determination are available for environmental




 analysis.  However, sample-handling may present difficulties because of losses




 of arsenic compounds via sublimation, especially during air monitoring, and




 analytic personnel should be alerted to this possible procedural pitfall.




      Individual arsenic compounds can be determined only after isolation by




 an appropriate method, such aa volatilization, paper chromatography, gas




 chromatography, or electrophoresis.




                                                                       When




 the nature of the compound is known, the quantity present can be measured by




 measuring the amount of arsenic present.




      The continued concern about the association between inorganic arsenic




 and cancer has raised questions regarding the implications of widespread




dispersion of inorganic arsenicals in the environment.  Clearly, the ecologic




 uncertainties about arsenic compounds deserve more effort and attention.
                                     -332-

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                                CHAPTER 8
                             RECOMMENDATIONS
     Review of the scientific literature by the Subcommittee on Arsenic
identified several subjects on which additional information is needed.   If
the following recommendations for research are successfully carried out, the
new knowledge thereby generated should allow a more accurate assessment of
the environmental impact of arsenic compounds.
1.   Further epidemiologic and laboratory experimental research should be con-
     ducted on the question of the possible carcinogenicity of arsenic.
     The possible carcinogenicity of arsenic remains controversial, and it is
     urged that more studies be done to settle this issue.  A group of experts
     should be convened to address this question specifically and to recommend
     and oversee studies in man and experimental animals designed to resolve
     the enigma.  Experts in the following subjects should be on the working
     group:  pathology of cancer; epidemiology; statistics; chemistry,  bio-
     chemistry, and metabolism of arsenic; and experimental design (especially
     as the last relates to the many possible confounding factors that may
     modify carcinogenesis).  Numerous opportunitities exist for additional
     epidemiologic work, and follow-up studies should be performed on popula-
     tions that have been inadvertently exposed to arsenic.  The problem of
     experimental arsenic cancer in laboratory animals also requires more effort,
     and a series of studies designed as rationally as possible should be carried
     out, to determine whether arsenic can be demonstrated to be a carcinogen
     under experimentally controlled conditions.  In such studies, careful
     attention should be given to various experimental characteristics, such
     as the species of animal, the dosage of arsenical administered, the nature
     of arsenical tested, the duration of arsenical exposure, and the route of
     exposure to the arsenical.   Possible cocarclnogenic effects of arsenic
     compounds with other chemicals should also be considered.
 The term "arsenic" is used here as a general term for arsenic compounds.
                                      -333-

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2.    More research is required to clarify the effects of long-term low-dose




     exposures to arsenic on man, domestic animals,  wildlife,  and aquatic.




     organisms.




     Recent studies using sensitive indicators of biochemical toxicity, such




     as alterations in enzyme activity, or physiologic criteria of poisoning,




     such as impaired reproductive performance, have suggested subtle changes




     at exposures of arsenic that were previously thought to be innocuous.




     How pertinent such results are to environmental problems is not certain,




     but at least the preliminary experiments should be confirmed or refuted




     and an attempt made to put such experiments into perspective.







3-   Additional studies on the possible teratogenic and mutagenic effects of




     arsenicals need to be carried out.




     All experimental teratology studies that have been carried out with




     arsenic compounds have used doses far in excess of those likely ever to be




     encountered as a result of environmental contamination.  Research with




     more realistic doses should be encouraged, to evaluate whether arsenic in




     the environment actually constitutes a teratogenic risk.   Experiments carried




     out with humans previously treated medically with arsenicals revealed




     chromosomal abnormalities, which suggest a mutagenic potential for some




     arsenic compounds.  Again, however, the doses of arsenic given to patients




     in the past were higher than any reasonable degree of environmental arstaic




     exposure that one would expect.   But the positive results argue strongly




     for further work along these lines.
                                    -334-

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 4-   Much more effort is required regarding the inhalation toxicology of
      arsenic.
      The physiologic significance of some of the experiments in this field
      is open to debate, but the observation that biologic changes occur under
      certain conditions apparently is not.  Alterations in metabolic or
      biochemical characteristics are observed at exposures that seem very low.
      This work needs to be repeated, and any possible physiologic relevance
      of these data needs to be pointed  out.


5.    Possible metabolic  interrelationships of arsenic with other pollutants
      should  be  explored.
      Metabolic  antagonisms  between arsenic and  some minerals suggest that
      arsenic may have antagonistic or synergistic effects with  other pollutants.
      This  illustrates the  fact  that  environmental standards for pollutants  can-
      not  be  set  in isolation,  but should take into account possible  interactions
      among pollutants.
 6.    The  use of the rat  as  an  experimental animal in  studies of arsenic meta-
      bolism  should be  strongly discouraged.
      The  rat has a unique  arsenic metabolism that is  totally unlike  that  of
      man  or  other  mammals.   Therefore,  research conducted with  rats  is difficult
      to apply to man; such  research  has led  to  many misinterpretations.   One
      such misconception is  the idea  that arsenic is retained in the  body  to the
      same extent as heavy  metals, such  as lead, mercury, and cadmium.
                                   -335-

-------
7.    More information about the chemical nature of arsenic in soil, water,
     foodstuffs, and plant and animal tissues is desirable.

     The behavior of arsenic in the food chain cannot be fully understood with-
     out increased knowledge of the various chemical forms of arsenic.
     The data are very incomplete, although it is clear that the naturally™
     occurring arsenic in foods is metabolized quite differently from inorganic
     arsenicals.  The recent attempts to characterize the arsenolipids in marine
     oils show what can be accooiplished in this direction, but more effort is

     indicated,   The forms of  arsenic in foods have  unknown toxicities  and

     environmental  behavior.   Further examination of their identity,  toxielty,
     and  fate in the environment  is needed,  so that  their  significance  to
     both man and his  environment  can be assessed.

8.   Better analytic techniques and  sample-handling  procedures for...arsenic
     compounds need to be developed.
     Most current analytic techniques for arsenic give values only for the total
     amount of arsenic in the  sample and do not characterize the various chemical
     forms  of arsenic present.  Because the toxicity and ecologic behavior of:
     arsenic depend strictly on its  chemical forms,  means to identify these izorms
     are needed.  Recent evidence also suggests that the equilibrium vapor
     pressure of some arsenic  compounds (e.g., arsenic trioxide) is great enough
     for appreciable losses to occur as a result of sublimation when dust
     particles are collected on high-volume air samplers.  Sublimation losses
     may also occur during sample storage or drying.  Surveillance personnel
     need to be alerted to these possible problems of analysis, and alternative
     procedures may have to be worked out.  Once acceptable methods for the
     determination of arsenic  compounds are established, routine monitoring of
     arsenic in environmental  samples should be undertaken.
                                  -336-

-------
 9.    An economic assessment  should  be  made of  the  possible  effects  that  not




      using  arsenical  pesticides  would  have on  food and  fiber  production.




      The organic arsenical pesticides  play an  important role  in protecting crops




      and livestock from damaging pests.   Current usage  is estimated at 15,000-




      20,000 tons a year.  Loss of these  pesticides or major price adjustments




      due to low  availability of  starting materials (arsenic trioxide)  could have




      a  major economic impact on  American agriculture.   It is  urgent to assess




      the domestic and foreign consequences of  the  loss  of these compounds.




10.    Guidelines  on the disposal  of  arsenical wastes should  be developed.




      Arsenic is  an unavoidable byproduct of smelting operations.  It must be




      used,  stored, or disposed of safely.  For pesticides,  the safe disposal




      of containers is important.  Slag  from smelting operations,  as well as




      the arsenic trioxide that is collected, must  be used or  disposed of in  an




      acceptable  manner.  Perpetual  storage should  most  likely be avoided.
11.   Additional work is needed to elucidate^ the biochemical mechanisms res-




      ponsible for arsenic poisoning.




      Although the toxic effects of trivalent arsenicals are accounted for




      reasonably well on the basis of their reactivity with sulfhydryl groups,




      the mechanism of action of pentavalent arsenicals, both organic and




      inorganic, is much less understood.  Careful metabolic studies should be




      carried out to determine whether pentavalent arsenicals are reduced to




      trivalent arsenicals in vivo and, if so, to what extent.
                                    -337-

-------
12.   Experiments should be carried out to establish whether animals can adapt




     to the toxic effects of arsenic.




     It seems to be well documented that microbial systems can adapt Lo toxic




     concentrations of arsenic,  although the precise molecular mechanism of




     this effect is unknown.  Recent results that indicate that mammals also




     can adapt to arsenic to a limited extent should be followed up, and addi-




     tional work along these lines should be encouraged.






13.   The possible effect of arsenicals in decreasing the ability to resist




     infection needs to be investigated further.




     The mechanism of this effect of arsenic, inhibition of interferon forma-




     tion or action, is of both theoretical and practical interest.  If this




     work can be verified and is found to hold true in other species, the




     implications for public health could be considerable.






14.   Arsenic should be studied as a possible nutritionally essential trace




     element.




     The occasional favorable effects of arsenic in animal metabolism suggest




     that it may play a physiologic role at very low concentrations.  Such a




     role has recently been shown in experiments using modern techniques in




     trace element research, but these studies need to be verified and expanded.
                                   -338-

-------
15.  The mechanism of action of arsnical "growth-promoting" agents should be




     studied.




     Although many theories have been advanced in an attempt to explain the




     growth-promoting effect of organic pentavalent arsenicals, none of these




     hypotheses seems totally satisfactory.   If these compounds are to be con-




     tinued in use, a better understanding of their mode of action might allow




     the design of equally active yet less toxic compounds.






16.  Studies on environmental characteristics that can affect the redistri-




     bution of arsenic within the ecosystem should be undertaken.




     Environmental conditions can seriously affect the toxicity of arsenical




     residues in soils.  Dissipation of applied arsenicals is subject to




     changes in rate and is a function of the environment and the arsenical.




     Information is needed on how these dissipation rates can be changed to




     prevent the buildup of toxic residues.






17.  An estimate of annual arsenic usage in agriculture is needed.




     The exact annual production, distribution, imports, exports, and inven-




     tories of arsenical pesticides is unknown.  Furthermore, it is impossible




     with current estimates to predict market trends as influenced by shortages




     in petroleum-based feedstocks, development of new pesticides, or any other




     economic change.  Consequently, the short- or long-term environmental




     impact of continued arsenic usage on agricultural production cannot be




     determined.
                                   -339-

-------
                 APPENDIX A




ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS




            Arsenic Concentration, ppm
     Treat-
(dry wt)
Plant or Product
CEREALS
Wheat



Wheat flour
Corn, grain



Corn, stalk and leaves




Corn, seedling
Corn, pop
Barley


Barley, straw
Rye
ment3-
S

SP
6S
7S


1
6S
7S

3
6S
7S
IS
3S


S
SP
SP

Treated
14.6

trace-0.0-
0.09-0.30
0.06-0.50


0.04-0.07
«0.05-0.8
<0. 05-0. 19

0.09-17.1
1.82-3.75
0.2504.36
2.76
1.8-252


4
0.0-0.9^
14. 3^

Nontreated

0.007-0.3

0.09-0.16
0.09-0.16
0.01-0.09
<0.01-0.4
<0. 01-0. 05
0.05-0.07
<0. 05-0. 10
0.04^
0.6-2.5
1.83-1.90
0.10-1.94
0.71
3.0
0.1
<0.1-0.55



<0.1
Reference
216
39,81,110,362,653,691,
740,771
322
19a
19a
34,38,371,691
81,110,353,364,740,771
125
19a
19a
353
780
19a
19a
364
783
110
110,353,362,653,771
71
322
322
653
                  -340-

-------
                 APPENDIX A




ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS




            Arsenic Concentration, ppm
     Treat-
(dry wt)
Plant or Product
Oat


Straw
Millet t
Rice, grain
Rice
Rice, whole plant
Bread
Whole wheat
Ginger
VEGETABLES
Soybean


Soybean, fodder

Soybean, oil
Soy sauce
Beans, green




mentlL Treated

IS 0.74-1.03
SP 0.1-
1S 2.07


S 0.5-5.0
2 0.9-9.4





6S 0.05 1.31
7S 0.06-2.03
6S 0.09-2.44
7S 0.48-3.08

100^-

IS trace-0.22
SP traced
3S 1.2-28.5
4S trace-26.6
N entreated
<0.1-2.28
0.09-0.13

0.28
<0.1
<0. 07-3. 53
0.4
0.8-5.0
0.016-0.03
0.008-0.02
0.05-0.07

0.08
0.05-1.22
0.05-1.22
0.07-2.12
0.07.2.12
0.09

0.1^-0.4
0.01-0.08

trace
trace
Reference
274,353,362,653
124
322
124
653
38,81,353,371,653,689
596
208
461
461
461

371
19a
19a
19a
19a
81
498
38,110,353
124,125,351,479
322
132
132,351
                  -341-

-------
APPENDIX A
ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS
Arsenic Concentration, ppm
Treat- (dry wt)
Plant or Product ment£.
Beans, pod
Beans , leaves
Beans
Bean
Bean, vines
Bean, roots
Bean, kidney
Bean, lima
Bean, yellow eye, leaf
Bean, black wax, leaf
Bean, black wax
Pea

Peas , pod
Peas, vine
Peas, root
Peas, sweet

Peanut
Carrot


Carrot, tops
Carrot, roots
IS
IS

N
IS
IS


IS
IS
is

IS
IS
IS
IS
SP



IS
SP
IS
IS
Treated
0.79
1.92

4.50
1.82
5.78


0.25-3.00
4.58
0.25

0.04-0.48
0.88
2.04-5.70
1.20
trace-0.1—



trace-0.27
0.0-2.91
0.57
0.18
Nontreated
0.27
0.21
0.05-0.40
0.07
0.18
0.29
0.33
0,4
0.08-1.14
1.57
0.08
<0. 01-0. 49
0.01-0.40
0.05
0.12-2.82
22.70

0.3
0.01-0.30
0.03-0.80
<0. 01-0. 08

0.00-0.57
0.32
Reference
351
351
479,771
451
364
364
368
110
124
124
124
353,364,479,653,740
364,455
364,455
364,455
364
322
110
81,740,771
81, 110, 123, 353, 479, 65j, 771
124,125,479
322
81,364
364
-342-

-------
                 APPENDIX A




ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS




            Arsenic Concentration, ppm
     Treat-
(dry wt)
Plant or Product
Potato




Potato, peelings
Potato, sweet
Onion


Onion, tops
Turnip

Turnip , greens
Rutabaga
Parsnips
Beets, tops
Beets, roots

Beets
Radish


mentf.

IS
SP
5S
4S
AS


IS
SP
IS

IS



IS
SP
IS
IS

SP
iS
Treated

0.06-0.11
0.0-0.1-^
Sl.O
trace-0.6
0.2-83.0


0.16-0.36
trace-3.2^
8.85

0.10



0.08-1.46
0.0-0.4^.
0.08-1.28
20.2

trace
0.02-0.22
N entreated
0.0076-1.25
0.01-0.05

0.2
trace-0.1
0.4-2.4
0.00
0.015-1.54
0.08-0.36

3.19
0=036-0.83
<0.01
0.03
0,80
0.20
0.07-3.48
0.1-1.3
0.34
1.27
0.01-2.02


Reference
81,110,353,369,651,740,
771
124,125
322
657
351,516,678
678
81
110,353,368,479,740,771
124,364
322
364
125,353,771
125
81
771
110,771
124,364,479
110,364,368,479,651,771
364,377,479
364
81,110,322,353,368,479,
771
322

                   -343-

-------
                 APPENDIX A




ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS




            Arsenic Concentration, ppm
     Treat-
(dry wt)
Plant or Product
Tomato




Tomato, stem and

Tomato, root

Eggplant

Eggplant, roots
Cucumber

Pickle, sweet
Lettuce


Lettuce, roots
Parsley
Watercress
Spinach
Kale

mentfL

IS
4S
3N
4N
leaves 4S
IS
4S
IS

IS
IS

IS


SP
IS
IS




IS
Treated

f:ra"ce-0.09
0.68-39.5
3.75-145,28
trace-18.1
-334
LI. 4
-1,707
1.93-12.83

trace-19.68
9,84

0.2


0.0-2.1-
0.08-0.32
10.98




0.98
N entreated
0.01-2.95
0.08-0.09
<0.2
trace
trace
<0.2
6.75
<0.2
0.26-0.49
0.18-0.77
0-6.14
0.98
0.02-2.4

0.14
0.01-3.78

0.12
0.47
0.1-8.0
1.84-2.10
0.04-2.25
0.11-0.22
0.01-0.99
Reference
81,110,368,651,771
124,364,479
132,250
132
132
250
364
250
364
368,651
479
364
81,368,479.651,751
751
110
38,81,353,364,479,740
322
364,479
364
110,227,248,771
353,368
81,353,368
409
81,364
                   -344-

-------
                 APPENDIX A




ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS




            Arsenic Concentration, ppm
     Treat-
Cdry wt)
Plant or Product
Kale, roots
Swiss chard
Kohlrabi
Cabbage
Chicory

Lentil

Celery
Celery, whole plant
Celery, stalks
Celery, root
Salsify
Asparagus
Mushroom, canned
Broccoli
Cauliflower
Endive
Pepper

Pepper, roots
Squash
uent*. Treated Nontreated
IS 17.49 0.39
IS 0.04-0.27 <0. 91-0. 08
SP trace-O.lk-
0.0-2.01
0.62
0.1*
0.70
0.3*
0.2-0.75
2.32
0.60
IcOO
0.11^
0.1
0.45-0.79
IS trace
0.86
0.21
IS trace-0.47 0.39
0.00
IS 6.89 1.57
0.023-0.034
Reference
364
124,125
322
81,110,322,353,368,651,
771
38
353
38
353
110,123,353
353
771
123
353
353
353,461
479
353
740
364,479
81
364
479
                 -345-

-------
                 APPENDIX A




ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS




            Arsenic Concentration, ppm
     Treat-
(dry wt)
Plant or Product
FRUITS
Apple


Apple, skin
Apple, butter
Apple, juice-cider

Orange
Orange, juice
Pear

Pear, skin
Peach
Peach, leaves
Apricot
Lemon
Lemon, leaves
Lemon, roots
Pineapple
Banana
Pumpkin
mentJL Treated Nontreated

0.04-1.72
SP trace-0.1-
1 0.33-14.2 0.03-1.91
0.07.1.35
0.43-2.41
0.065-0.165
1 0.18-0.47
0.11-^-0.35
0.008-0.12
0,17^-0.39
1 4.0
0.40-0.60
0.07-1.5
S 1.38-2.39
0.15-1.5
0.50
3N 11.0 0.35
3N 1,200
0.08^.
OoOob
0.09^
Reference

110,304,651,675,709,740,
771
322
68,353,602
675,700
110
675

38,353,492
492
304,353
70
700
81,134,740
44
134
38
419
419
353
353
353
                  -346-

-------
                 APPENDIX A




ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS




            Arsenic Concentration, ppm
     Treat-
(dry wt)
Plant or Product
Blueberry
Blueberry, leaves
Blueberry, stems
Blueberry, roots
Grapes


Grape, leaves
Grape, juice
Grapefruit , leaves
Mandarin
BEVERAGES
Wine




Wine, white
Wine, red
Wine, fruit
Wine, port
Lemonade
Whiskey
Beer
ment*- Treated Nontreated
2S <1.4-4.3
2S 6.74-14.97 0.78
2S 7.6-13.3 0.27
2S 93.7-164.2 2.40
0.75-1.20
6S 0.24-0.28 0.05
IS 0.6-3.8
2.3
None Detected
2.0-3.0
0.85

0.005-0.15
IS 0.0-4.0
2S 0.4-1.0 0.01-0.02
2 2.76 <0.1
1 0.02-0.18
0.06-0.56
0.03-1.38
0.06-0.11
0.02-0.07
0.000-0.005
0.02-0.07
0.01-2.0
Reference
43
13
13
13
700
169
311
314
314
612
353

80,461
251
212
575
738
133
133
165
262
461
80
80,461,470,743
                 -347-

-------
                 APPENDIX A




ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS




            Arsenic Concentration, ppm
     Treat-
(dry wt)
Plant or Product mentS. Treated
Ale
Malt
Hops, sundried
Hops, sulfured 0.15-19.5

Coffee, bean 1 0.5-1.5
Liquid fruit products
TREES
Pine, short leaf
Fir, Douglas, needles
Fir, Douglas, twigs (ash)
Fir, Douglas, twigs AM
Cypress, Italian
Fir, (Abies alba)
Pine (Pinus laricius)
Chestnut (Castanea vesca)
Pine, Scotch
Beech, European
Spruce, white
Spruce, black SP
Oak, chestnut
Oak, chestnut, acorns
Oak, chestnut, roots
Hickory
Poplar, tulip
N on treated
<0.02
0.26-0.35
0.08-0.15

0.03-0.82

0.09-0.21

0.1-0.2
4-8000
<100
>1,000
1.4
0.11
0.13
0.05-0.11
<0.,05
<0.05
1.0-2.4
<1.0-96
0.05-0.40
0.1
11.0
0, 10-0. 40
0.08-0.40
Reference
470
470
134
134,423,681
422,423,681
569
165

17
172
749
749
495
495
495
353,495
495
495
605
401 b
17
17
17
17
17
                   -348-

-------
                 APPENDIX A




ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS




            Arsenic Concentration, ppm
     Treat-
(dry wt)
Plant or Product
Walnuts
Walnut, black
Hazelnuts
Date
Filbert
Cherry, leaves
Almond
Hemlock, foliage
FORAGE CROPS
Grass





Clover

Clover , red

Clover
Clover, white
Hay
Alfalfa




mentS.









IS
S
SP
4
1,5

IS

AS
S
IS
IS

SP
S
IS

Treated





8.60
2.0-4.8


0.5-2.0
0.5-75.5
2.5-12.0^
938-1,462
15,000-60,000

1.32
12.0
0.09-0.84
12.0
6.24
1.26

004-5.7^
14 .,0-860
3.37
-349-
N on treated
0.07
0.13^
0.78
0.12^-
0.11

0.3
0.2-0.4

0.1-0.7
0.50-0.94
0.5



<0. 10-0. 17
0.46
0.37
0.11-0.39

3.64
0»52
0.05-3.38


1*97

Reference
38
353
38
353
353
424
353
659 a

31
110,124
216,771
322
527
95
509
124
110,353,771
655
771
364
124
353,509
322
110,728
364

-------
                 APPENDIX A




ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS




            Arsenic Concentration, ppm
     Treat-
(dry wt)
Plant or Product ment£ Treated
Alfalfa, roots IS 0.78
Sudangrass
3N 9.6-384.8
4N 3.1-68.4
4S 0.38-33.4
Vetch IS 1.92
Vetch, roots IS 15.82
Sunflower
Sunflower leaf SP 3.3^
MARINE ALGAE, SEAWEED
Macrocystis pyrifera
Chondrus crlspus
Laminaria digitata lamina
Laminaria digitata
Laminaria digitata, oil
Laminaria digitata, fatty acid
Laminaria saccharina
Laminaria saccharina, oil
Laminaria saccharina, fatty acid
Halidrys siliquosa
Fucus nodosus
Entarompha compressa
Ahnfeltia plicata
Fucus vesiculosus
Fucus vesiculosus, oil
Fucus vesiculosus, fatty acid
N entreated
3,15
0.70
trace
trace

1.22
7.15
<1. 0-2.0


4.0-60.0
3.8-18.0
107-109
47.0-93.8
221.0
36.0.
45. OF- - 52.5
155
7.5-52o5
26.0-30.0
45.0
11.2
39.0
24-65
35.0
5ol
Reference
364
451
132
132
132
364
364
110
322

264,771,
121,361,
443
361,409
438
438
361,409
438
361,409,
361,409
361
361
409
361,443
438
438










773
409
438


                   -350-

-------
                 APPENDIX A




ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS




            Arsenic Concentration, ppm
Treat- (di
Plant ot i'roduct ment5 Treated
Fucus serratus
Fucus serratus, oil
Fucus serratus, fatty acid
Piocamium coccineum
Ulva latissima
Gigartina mammillosa
Laminar ia hyperborea, oil
Laminaria hyperborea, fatty acid
Ascophyllum nodosum, oil
Ascophyllum nodosum, fatty acid
Fucus spiralis
Fucus spiralis, oil
Fucus spiralis, fatty acid
Pelvetia canaliculata
Pelvetia canaliculata, oil
Pelvetia canalicilata, fatty acid
ALGAE
Algae
Alj^ae , Odegodeum 7 4.5-71.4
Sceletonema costatum, oil
Chlorella ovaliSj on
Chlorella pyrenoidosaj Oil
Phaedactylum tricornutum 0-n
Oscillatoria rubescens On
Pterygophera californica
Agarum fimbuetum
Rhodemia pertusa
Casteria castata
•y wt)
N entreated
28-67.5
27.0
6.1
7.5
6.0
4.5-17.2
197
16
7.8-49.0
5.2-21.0
15-34
5.7
5.0
15-22
10.8
7.3
0.5-12.0
1.3
0.7
0.5
3.6-4.8
Oo4-0.5
12oO
4.0
1.0

Reference
361,443
438
438
361
361
362,443
438
438
438
438
443
438
438
443
438
438
110,465,771
347
441
441
441
441
441
771
771
771
771
                   -351-

-------
                 APPENDIX A




ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS




            Arsenic Concentration, ppm
     Treat-
(dry wt)
Plant or Product menti Treated
MISCELLANEOUS
Baking powder
Cottonseed 6 0,05-0.08
Cottonseed products 5 On68

Cotton leaves 6 0.13-41.4
Sugar
Glucose
Honey
1 0.9
Pectin
Organic food color
Gelatin
Polyphosphates
Chocolate
Jam
Jam and marmalades
Tobacco
Johnson grass
Sugar cane
AQUATIC PLANTS, NEW ZEALAND
Ceratophyllum demur sum GTA 20-1060
Lagarosiphon major GTA 29-1450
Nontreated

1.0
0.05

0.58
0.055
0.15
0.2
0.14

1.0-3.55
3.0
1.0
<0. 3-3,0
0.07-1.53
0.00-0.1
0. 04-0 o 08
trace-42.8
Oo65
2,,0

1.4

Reference

34
36
81
400
769
38
461
110
197
87,110
87
87
87,579
38,461
461
165
110,282,428,519,662,792
400
563

401,594,595
401.594,595
                  -352-

-------
                 APPENDIX A

ARSENIC CONTENT OF PLANTS AND PLANT PRODUCTS

            Arsenic Concentration, ppm
     Treat- 	(dry wt)
Plant or Product mentS. Treated
Elodea canadensis GTA 307-700
Potamogeton sp. GTA 45-436
Lemna sp. GTA 30
Nitella hooker i GTA 182
A. Braun
Enteromorpha nana GTA 14-40
Compsopogon hookeri GTA 550
Typha orientalis Presl GTA 8
Egeria densa GTA 266-310
Atriplex confertifolia
Myriophyllum propinquum GTA 456
MISCELLANEOUS
Mustard, paste
Rhubarb
Astragalus bisulcatus
Russian thistle
Turpentine weed
Wild aster
Ironweed
Ragweed
Tulipa sp.
Scarlet mallow
Oreocarya sp.
Stanleya pinnata
Cocklebur
Spurge
Nontreated Reference
3.0 401,595
<6.0 401,595
2.5 401,595
13.0 401,595
401,594,595
594,595
401,595
3.2 304
— 401

0.28 771
<0.1 771
2.0 771
<1.0 771
<1. 0-2.0 771
2.0 771
1.0 771
1.0 771
2.0 771
loO 771
-------
                                    APPENDIX A


                   ARSENIC CONTENT OF PLANTS AND PLANT  PRODUCTS


                               Arsenic Concentration, ppm
                        Treat-
(dry wt)
Plant or Product
Aplopappus fremontii
Astragalus pectinatus
Lambsquarters, common
Mustard , common
Clintonia borealis
Sorrel
Dandelion
Wild leek
Buckhorn plantain
Sourgrass
Daisy
Milkweed , tops
Sour dock, tops
Burdock, leaves
False indigo
MOSS
Hylocomium splendens
Pleurozium schreberi

ment£.



S
SP
S
S
S
S
S
S
S
S
S
SP
SP
Treated



<1.0
<2-52
7.0
8.0
16.0
18.0
12.0
1.0
1.0
-------
                                  APPENDIX B

                          ARSENIC CONTENT OF ANIMALS
                                     Arsenic concentrat-
                                     lon, ppm  (fresh wt)
Hair

  Distal

  Proximal
Brain
Teeth

Esophagus

Thyroid
Lung
  Female

Heart
KpO-
urea



1
2

3
4

4


2
3
4


4


Exposed Normal
0.3-1.75
0.79
0.03^-1.92
0.4-816 <3.0
3.58 0.997
0.001—0.14
1.0-1.4
1.9
0.003-0.635
168
0.06-0.13
0.001-0.314^.
0.003-0.332^
0.002-0.093
0.08-0.17
2.3-2.6
20.0
0.006-0.514^
0.006-0.038
64.0
0.002-0.078^.
0.001-0.016
References
80,123
253
80,253,370,472,
666,730
103,189,491,585
47
253,370,666
272
480
666,733
480
61,253
666
666
393a
253,370
272
480
666
501
480
666
253,370,761
                                      -355-
-------
        APPENDIX B

ARSENIC CONTENT OF ANIMALS
           Arsenic concentrat-
           ion, ppm (fresh wt)
Animal
Liver



Kidney

Left
Right

Pancreas


Bladder
Gall Bladder
Stomach

Walls
Contents

Intestine
Small
Large
Expo-
sure8 Exposed Normal References
0.09-0.30 80,253,370
3 4.4-6.9 272
4 12.8-143 480
0.005-0.246^ 666
0.07-0.14 253,370
3 0.4-1.3 272
4 15.8-92 480
4 81 480
0.002-0.363^ 666
0.07 253
4 94 480
0.005-0.410^- 666
0.06 253
4 41 480
0.04 253
3 0.1-0.3 272
4 5-246 480
4 5-8,836 480
0.003-0.104^ 666
0.07 253
4 132 480
4 259 480
            -356-
-------
                                  APPENDIX B

                          ARSENIC CONTENT  OF ANIMALS
                                     Arsenic  concentrat-
                                     Ion,  ppm (fresh wt)
Bone

  Calvarium

  Rib

Nail
Blood
  Women, venous

  Menstrual

  Serum

  Skin

  Spinal cord

  Urine
xpo-
ure* Exposed

3 0.5-2.2
4 12.8





5 7.1-17.8

4 20-130
6 0.82-3.0

4 5.0

7 0.03-0.27




4 20.6
1 0.04-0.9
Normal
0.08-0.13


0.001-0.132^-
0.16-0.50
59-61 (in ash)
20-27 (in ash)
1.70
0.04-0.11
0.02-2.90^-


0.01-0.59

0.001-0.920^
0.01-0.13
0.06-1.44
0.18
0.000-0.0028
0. 009-0. 59^

0. 01-0 o 22
References
61,253,370
272
480
666
253,370
538
538
253
344
666
731
183
253,281,472
480
666
744
281,742
742
168,472
666
480
103, 356,491,558
                                      -357-
-------
                                    APPENDIX B

                            ARSENIC CONTENT OF ANIMALS
                                       Arsenic  concentrat-
                                       ion, ppm (fresh wt)
Animal
                               Expo-
    Uterus

      Membrane

    Aorta

    Adrenal

    Breast

    Muscle, pectoral

    Ovary

    Prostate

Domesticated Animals

Beef

  Calf muscle

  Calf liver

  Liver

  Milk



  Milk, dried

  Milk, sterilized

  Milk, condensed

  Butter

  Veal
Exposed Normal
0.000-0.11
27.2
0.010-0.188^
45.6
0.003-0.570^
0.002-0.293^-
0.030-0.221^-
0 o 012-0. 431-
0.013-0.260^-
0.010-0.090^-
0.008
0.52
0.15
0.063
0.0005-0.07
«0. 05-0. 27
<0o5
0.03-0.04
0.01-0.014
0.07
0.005-0.010
References
253,472
480
666
281
666
666
666
666
666
666
39
580

547
39,372,461
216,467
702
461
461
38
39
                                         -358-
-------
                                   APPENDIX B

                           ARSENIC CONTENT OF ANIMALS
                                      Arsenic concentrat-
                                      ion, ppm (fresh wt)
Animal
Pork
Muscle
Muscle
Salami
Chicken
Meat
Kidney

Liver

Liver
Bone, marrow
Eggs
Yolk
Rabbit, muscle
Liver
Kidney
Heart
Lungs
Meat , canned

Fat
Wild Animals
Expo-
sure8 Exposed Normal

0.22-0.32
7 0.29-0.92 <0.02
0.14-0.20
7 0.2-1.2
7 0.01-0.4 0.02
7 0.08-1.2 0.05
0.02
12.2
0.02
7 0.19-2.43 0.08
0.0-0.4
7 0.40
0.005
7 1.0-2..0
7 3.0
7 3.0
7 trace
7 trace
Oo 01-0. 18
0.20-4.13^-
0.13-0.54-

References

38,580
411
38,110
140
40,140
40,140
252
140
252
40
140
210
39
652
652
652
652
652
580
653
653

Squirrel
0.8
17
                                       -359-
-------
        APPENDIX B

ARSENIC CONTENT OF ANIMALS
           Arsenic concentrat-
           ion, ppm (fresh wt)
Expo-
Animal sure3 Exposed
Sparrow
Mice
Hawk
Owl
Fox
Crow
Oppossum
Starling
Bee, dead 4 3.22-12.0
2 20.8-31.2
Pollen 2 Lrl20
4 7.75-30
Larvae, dead 4 4.95-13
Pupae 4 10
Honey 4 16-22
2 1-2
Aquatic Organisms
Shrimp
English potted
Edible portion
Canned
Pandalus borealis, oil
Pandalus borealis, fatty acid

Normal
0.2k
1.0*
0.4k
0.05k
o.sk
0.1*
o.2k
0.01-0.21








1.27-41.6
8«2-18.8
0.95-31.2
0.08
10.1
4»8
13. 0-42. G£
References
17
17
17
17
17
17
17
470
197,450
430
697
197,199
197,199
199
199
430

110,121,142
121
143
174
438
438
444,450
             -360-
-------
                                     APPENDIX B

                             ARSENIC CONTENT OF ANIMALS
                                        Arsenic concentrat-
                                        ion, ppm (fresh
Animal

  Palamon serratus, cooked

  Parapeneus longirostris

  Crab

    Dressed

    Canned

    Muscle



  Carcinus maenas, cooked

  Cancer pagurus, cooked

Clam, minced



  Canned

  Pecten maximus

  N-liquor

  Oil

  Fatty acid

Prawns

  Dublin Bay

  American tinned

  Japanese tinned

Oysters

  English

  Portuguese
                                Expo-
Exposed Normal
1.0-2.7
1.7-38.2
27.0-52.5
18.8-62.6
0.71
6.1
S7.95
2.5-7.0
2.1-33.4
0.85
1.42-2.56
0.36
11.6
18.0
4.8
1.9
34.1
27.0-130.5
10.5-30.0
15.0-63.8
0.3-3.7
2.2-7.5
24.8-5205
References
141
141
121
121
174
334
708
141
141
110
110, 764
174
439
439
438
438
110
121
121
121
110, 147, 321, 454b, 764
121
121
                                       -361-
-------
                                    APPENDIX B

                            ARSENIC CONTENT OF ANIMALS
                                        Arsenic concentrat-
                                        ion,  ppm (fresh wt)
                                Expo-
  Ostrea edulis
    N-liquor

  Gryphea angulata

Lobster (Homarus vulgarus)

  Canned

  Fillet

  Fillet, N-liquor

  Cooked

  Muscle

  Whole

  Norwegian, cooked
  (Nephrops norvegicus)
Scallop

Mussel

  Mytilus edulis


  Mytilus edulis

  Whole

  N-liquor

  Oil

  Fatty acid
Exposed Normal
0.4-0.8
0.22
1.00
2.6-8.2^
9.8
1,2-3.6
2o27-54.5
0.94
5.3
14.0
10.8-17.2
0.022
00453
7.2-19.4
27oO-63«8
2.58-89.2
Oo08-8.0
9o 5-15.0^-
.ob
9.7
18.0
22.0
References
121
174
174
409,439
439
141
110,121,141,173
174
439
439
141
39
39
141
110,121
110,121,173,444
141,173,264,439,
450
409
444
439
438
438
                                         -362-
-------
                                     APPENDIX B

                             ARSENIC CONTENT OF ANIMALS
Animal

  Mytilus edulis, whole
        Arsenic concentrat-
        ion, ppm (fresh wt)

Expo-
sure*   Exposed      Normal
        0.04-0.09    o.Ol
References
628
  Mvtilus magellonicus

Cockle

  Cardium edule



  Cardium edule
                     10.5-26.1-
  Tapes decussatus

Whelk

  French edible snail

  Buccinum undatum

Periwinkle
  Littorina littorea
  Littorina littorea

  Littorina littorea, oil

  Littorina littorea, fatty acid

  Cooked

Crawfish

  Palinurus vulgaris, cooked

  Palinurus vulgaris

  Astacus pallipes

Squid

  Omnastrephes sagittatus
5.1-8.4
12.8-30.0
1.3-2.4
3.7-6.6
9.0-30.0
0.4
11.0H
12*
14. 0-19 oO^-
84,0
32.0
3.6-6.3
12.0-54.6
15 o 0-33. 8
Oo8-1.5
409
110,121
141
141
121
121
409
409
409
438
438
141
141
121
121
                     6.5
439
                                         -363-
-------
        APPENDIX B

ARSENIC CONTENT OF ANIMALS
           Arsenic concentrat-
           ion, ppm (fresh wt)
Expo-
Animal sure3 Exposed
N-liquor
Loligo vulgaris. raw
Loligo vulgaris, cooked
Fatty acid
Starfish (Asterlas rubens)
Starfish, oil
Starfish, fatty acid
Cuttlefish
Sepia officinalis, 'gills
Sepia officinalis, mantle
Sepia officinalis, raw
Sepia officinalis, cooked
Anchovy
Octopus, blood
Octopus bimoculatus, tentacles
Octopus vulgaris, raw
Octopus vulgaris, cooked
Cod (Gadus morrhua)
Fillet
Fillet, N-liquor
Muscle
Liver
Liver, oil
Black Marl in
Muscle
Liver
Normal
17.0
0.8-7.5
0.4-3.3
?67-
9.1
7.5

198^
73*
6.2-11.5
0,8-6.8
7.1-10.7
0.01
0.12
2.6-40.3
3.0-31.0
3.69-24.3^
2.2
13.0
0.4-0.8
0.7-3.2
1.4-10.0
0.1-1.65
0.1-2.75
References
439
141
141
438
409
438
438

409
409
141
141
446
488a
264
141
141
110,444
439
439
621
621
110,329,43*
621
454 ~
454 -
          -364-
-------
        APPENDIX B

ARSENIC CONTENT OF ANIMALS
           Arsenic concentrat-
           ion, ppm (fresh wt)
Expo-
Animal surea Exposed
Tuna
Tunny (Thunnus thynnus)
Haddock (Melanogrammus aegleflnus)
Mullet, red
Dogfish
Plaice
Fillet, oil (Pleuronectes plates sa)
Fillet, fatty acid
Sole (Solea solea)
Dab
Caviar, Russian
Pike (Esox lucius)
Pike 7 0.0
Perch (Perca fluviatllis)
Perch, yellow
Tench
Bream
Roach
7 $0.09
Trout, viscera 2 5.3
muscle 2 2«4

Normal
0.71-4.6
9.(£
5.54-10.8^-
Io54
0»53
4.5-7.5
6.1
5.2
5.2
2.2-3.0
3.8
0.8
0.0-0.11
0.6
0.06
0.4
0.4
0.4



0.069-0»149
References
110,547
444
110,444
38
38
121
438
438
121
121
121
121
599a,708
121
599 a
121
121
121
708
203
205
587
            -365-
-------
        APPENDIX B

ARSENIC CONTENT OF ANIMALS
           Arsenic concentrat-
           ion, pptn (fresh wt)
Expo-
Animal sure3 Exposed
Whitefish, viscera 2 3*6
Whitefish, muscle 2 2.7
Sucker, spotted, whole
Sucker, white
Shiner, golden
Bass, black, liver, oil
Bass, black, large mouthed
Large-mouth (Micropterus
Salmoides lacepede) 0.44-0.93
Salmoides lacepede, white
Carp
7 50.19
Catfish
Herring, fillet (Clupea harengus)
N- liquor
Meal
Oil
Muscle
Mackerel (Scomber scomber)
Meal
Fillet, N-liquor
Fillet
Fillet, oil
Fillet, fatty acid
Normal


0.062-0.253
0.11
0.55-1.95
7.37-77.31
0.01-1.86
0.08-1.20
0.28-0.48
0.055-0.51

0.07-0.298
3.8
6.4-24.0
2,7-6.9^
3»1-20.2
2.0
Oo027-9.2-
2.7-3.8
3<,2-17oO
2.2^-3.5
8.2
4ol
References
203
203
205,587
579a
205
205
205,587
205
205,579a
205,579a,587
708
579a, 587
439
439,447
437,444
438,442,446,621
621
444
437
439,447
409,439
438
438
             -366-
-------
        APPENDIX B

ARSENIC CONTENT OF ANIMALS
           Arsenic concentrat-
           ion, ppm (fresh wt)
Expo-
Animal surea Exposed
Liver, oil
Liver, fatty acid
Oil
Capelin, meal
Capelin, N-liquor
Oil (Mallotus villosus)
Fatty acid
Pout, Norway, meal
Pout, Norway, oil
Whale (Balaenoptera physalus)
N-liquor
Coalfish (Pollachius virens)
North Atlantic Finfish
Catfish Bagre Marinus
Eel Anguilla rostrata
Flounder Paralichthys lethostigma
Decapterus pujictatus
Normal
13.0
6,2
4.3-15.0
2.6-19.1
10.3
5.2-23.2
6.3
3.9
11.8
0U4
0.9
7.2*
-------
        APPENDIX B

ARSENIC CONTENT OF ANIMALS
           Arsenic concentrat-
           lon, ppm (fresh wt)
Animal

Eel (Conger sp.)

Anchovy (Anchova mltchelll)

Mullet(Mugil cephalus)

Hygophum hygomi

Ceratoscopelu warmingil

Notoscopelus caudispinous

Lobianchia dofleini

Lepidophanes indicus

Diaphus mollis

Lampanyctus pusillus

Ophichthus ocellatus

Ophichthus gomesi

Mo rone saxatilis

Sea trout (Cynoscion nebulosus)

Euthynnus alletteratus

Scomberomorus maculatus

Centropristes striatus

Coastal Organisms, England

Halichondria panicea

Tealia felina

Nereis diversicolor

Palaemon elegans
   Expo-
   sure8   Exposed
Normal
References
:1.0*
:1.0*
:1.0*
1.0*
:1.0—
1.0*
:1.0*
1.0—
1.0*
1.0*
1.0*
2.5*
1.8*
6.4^
2.8*
72.0*
16.0*
775
775
775
775
775
775
775
775
775
775
775
775
775
775
775
775
775
409
409
409
409
            -368-
-------
        APPENDIX B

ARSENIC CONTENT OF ANIMALS
           Arsenic concentrat-
           ion, ppm (fresh wt)
Expo-
Animal sure8
Patella vulgata
Crepidula fornicata
Nucella lapillus
Styela clava
Botryllus schlosseri
Anguilla anguilla, muscle
Marone labrax, muscle
Platichthys flesus, muscle
Shellfish, Portugal
Rock shell (Murex trunculus), cooked
Donax trunculus
Solen Marginatus
Cooked
Aristeus antennatus
Aolliceps cornucopia, cooked
MISCELLANEOUS
bluegills (.Lepomis macr ochirus )
7
Gar, long-nosed
Shad (gizzard)
Small-mouthed buffalo
Brook silversides
Drum, freshwater
Salmon, coho
Exposed Normal
11-24 ^
b
16. 0-38.0"
6.6^
ft 7b
o. / —
14.6-26.4
1.8-3.7
1.9-4.2
1.4-2.7
4.4-19.6
1.2-8.6
0.52
0.09-11.60
0.35-0.40
0.13-1.47
0.05-2.75
00 30-1. 26
0.09
0.09
References
409
409
409
409
409
409
409
409
141
141
141
141
141
141
205
256
205
205, 5793
205
205
579a
579a
            -369-
-------
                                     APPENDIX B

                             ARSENIC CONTENT OF ANIMALS
                                        Arsenic concentrat-
                                        ion, ppm (fresh wt)
 Animal

 Minnows

 Pickerel

 Black  bullhead

 Horned dace

 Gambusia

 Insects

 Cryptozoa

 Earthworms
 Snail
 Snail, garden

 Crustacea, planktonic

 Sea star (Pisoster ochraceus)

 Fish,  muscle

 Daphnia magna
Expo-
sure3
Exposed



0.07-11.20







3.9-254
Normal
0.14-1.95
0.13-0.73
0.22
0.42-0.65

1<£
10(£
b
0.3
3.2-5.5
102<£
3.06-6.8

References
205
205
205
205
347
17
17
17
347
121
205
264
334
347
—1 = industrial; 2 = pollution; 3 = AsH ; 4 = poisoned; 5 = As polyneuritis;

 6 = aerosol treatment; 7 = ted arsenic

—dry weight
                                         -370-
-------
                                APPENDIX C



            DETERMINING TRACES OF ARSENIC IN NATURAL MATERIALS







     This discussion is intended primarily for the consumer of analytic



information, i.e., for the physician, biologist, or ecologist who collects



and selects samples and wishes to obtain the most useful information from



them.  The principal paths by which arsenic can be accidentally added to or



lost from the system are mentioned, and the advantages and disadvantages of



the more commonly used analytic techniques are pointed out, so that the



investigator can choose among the available services and critically evaluate



the results.  The general approach is that followed in the recent review by


                  699
Talmi and Feldman,    although new material has been added and some of the



less accessible techniques omitted.



COLLECTION, SUBDIVISION AND STORAGE OF SAMPLES



     The sample collected should be large enough to represent the material



studied.  Because a single mean value is desired for the concentration of each



arsenical species of interest, the sample must be homogenized and a subsample



of suitable size for analysis must be taken.  To minimize contamination, unused



sample material should be stored in closed containers or (depending on sample



composition) at a low temperature.



     Choices must sometimes be made regarding what to include in the sample



taken for analysis.  Vegetation may be found to be contaminated with dust; a



decision must be made whether to remove the dust or include it in the sample.



Natural waters often contain suspended matter, which must either be filtered



out or allowed to remain.  If the particles filtered from an air stream contain



volatile forms of arsenic, consideration must be given to the losses that may
                                     -371-
-------
occur at the temperatures and air velocities to which the particles are



exposed on the filter and to the duration of exposure (arsenic trioxide has




a vapor pressure of 0.68 ran Hg at 200 C).



     Many authors (e.g., Portman and Riley;    Whitnack and Brophy;   a Al-


                lOa
Sibbai and Fogg;    and C, Feldman, personal  communication)  have found that




acidic, neutral, or basic solutions of inorganic arsenites and arsenates can




be stored without substantial changes in concentration for several weeks.




However, some arsenic compounds present in natural waters are said to disappear




rapidly from solution after collection of the sample (R. S. Braman, personal




communication).  The investigator must always be aware of the possibility of




losing some of the species of interest through adsorption on vessel walls or



on suspended matter or through volatilization.




     Large liquid samples can be properly divided into aliquots only if homo-



genous, i.e., if the species of interest does not adhere to the vessel walls




and if suspended matter is uniformly distributed before division.  Large solid



samples of a mineral nature may, of course, be subdivided by conventional




crushing or impact treatment followed by mixing and quartering or riffling.



Large samples of biologic tissues can be homogenized in a blender (with the



addition of water, if necessary).  If the density of the resulting slurry can



be stabilized  long enough, the sample can be subdivided in this manner.



Alternatively, the slurry can be centrifuged, and proportionate amounts of



residue and supernatant liquid taken for analysis.  Another possibility is




lyophilization of the slurry; the cake obtained is easily pulverized, and the


                                215b
resulting powder is homogeneous.     If volatile species are to be detert ined,




the lyophilization technique may not be appropriate.  The amount of handling




must always be minimized in order to minimize contamination.
                                    -372-
-------
PRETREATMENT AND DISSOLUTION OF SAMPLES



     If organic arsenic compounds are to be determined, the species in



question must be isolated.  '   a  If total arsenic is to be determined, the




arsenic must be brought into solution and, if necessary, converted to




inorganic form.  Regardless of the dissolution procedure used, care must be




taken to ensure that no arsenic is lost by the volatilization of trivalent




arsenic halides.  Loss can usually be prevented by boiling the sample with


                                                             699
concentrated nitric acid under reflux early in the procedure.




     The following sample preparation procedures are typical of those used in




environmental work.




     Coal is heated to fumes with concentrated sulfuric acid and treated with




successive small portions of concentrated nitric acid until degradation




essentially ceases.  Destruction of the remaining nitrogenous compounds is




completed by snail additions of fuming concentrated perchloric acid.  The




latter step is essential if the arsine generation-arc emission procedure is



                                            84
to be used for the final determination step.




     The arsenic in fly ash is usually assumed to exist as a surface coating.




All this arsenic can be dissolved with fuming sulfuric acid, as is shown by




comparison with analyses of the same material by neutron-activation analysis.



Refluxing such material in boiling water  for  1 hr recovers only 13% of the




arsenic present (C. Feldman, personal communication).  If the arsenic was




deposited from the vapor phase, it may have been thinly covered by other sub-




stances deposited  later.
                                   -373-
-------
     Coal slag is a highly refractory glass and usually contains only small



amounts of arsenic.  The arsenic that it does contain cannot be leached out




with ordinary acids.  Treatment with hydrofluoric acid in the usual way would




be of dubious value -- on the one hand, this reagent may contain substantial




amounts of impurities; on the other, arsenic trifluoride and especially arsenic




pentafluoride are rather volatile, so both contamination and losses might




occur.  Attack of the slag by fusion is open to similar objections.




     Quartz can be attacked without metallic contamination by vapor-phase



                                                                    793a
treatment with hydrofluoric acid arid nitric acid in a closed system.      This




approach was therefore tried with slag, albeit with some misgivings regarding




the volatility of arsenic fluorides.  No losses or contamination seem to have




occurred, however, inasmuch as the results obtained on fly ash agreed well with


                                                                  215c
those obtained with neutron activation and sulfuric acid leaching.




     Procedures for digesting plant or animal tissues for determining total




arsenic must completely convert the arsenic to inorganic form (preferably




arsenate) and must eliminate any substances that would interfere with the




particular procedure to be used in later determination.  Only the more widely




used digestion methods will be mentioned here; others have been reviewed


          699
elsewhere.     Small samples) can be charred with concentrated sulfuric acid


                                                                          127
and then subjected to repeated small additions of concentrated nitric acid


      A 9^ K
or 30%     or 50?0 hydrogen peroxide.  In the latter case, trivalent arsenic




will be lost if chloride is present.      Ordinary and fatty tissue weighing




up to 5 g can be safely wet-ashed in a volumetric flask by refluxing under a




short air condenser with appropriate mixtures of sulfuric, nitric, and


                                                         f\ -I c 1

perchloric acid, with potassium dichromate as a catalyst.
                                    -374-
-------
PRECONCENTRATION OF ARSENIC SPECIES



     To increase the sensitivity and accuracy of analysis, the arsenic-bearing



species is often isolated from its matrix and concentrated.  The principal pre-



concentration procedures used are coprecipitation, liquid-liquid extraction,



and volatilization.



     Coprecipitation with ferric hydroxide, Fe(OH)-, has long been known to



collect pentavalent arsenic quantitatively from solution at concentrations as



low as 2 ng/ml.   c>     The hydroxides of cerium and zirconium appear to be
                                             C QO „

effective as ferris hydroxide in this regard.      Thionalide can collect
                                                                             as
arsenic efficiently from comparatively large amounts of seawater,    but this



reagent apparently does not function well at low salt concentrations.



     Trivalent arsenic can readily be extracted from 6N hydrochloric acid with


                                            24 2a
mixtures of ketone and carbon tetrachloride.      At lower acidities, (pH, 2-6),



it can be precipitated with ammonium pyrrolidine dithiocarbamate and the pre-


                          515a
cipitate can be extracted.      If the arsenic is originally present in the



pentavalent state, this fact can be turned to advantage:  while the arsenic is



still pentavalent, other potentially interfering metals that are extracted



under the same conditions can be extracted and discarded.  The arsenic can then



be reduced and extracted without the metals that would otherwise have accompanied



it.



     Arsenic can also be separated from its matrix by volatilization, as arsine



(boiling point -55 C) or a substituted arsine.  The necessary reduction can be



effected by using zinc and acid in the presence of stannous chloride or


          , _,._,  219a,248a,456a        ,,  _,                          ,  ,
potassium iodide.                The reducing agent most commonly used, however,



is sodium borohydride, NaBH, .  The properties of this reagent can affect



analytic results, especially at low arsenic concentrations ( <. 1 ppm) , and will
                                  -375-
-------
therefore be discussed briefly.  Sodium borohydride is supplied commercially

in the form of 0.20- 0.25-g pellets or powder; the grade usually used for

analysis is the same as that used in preparative organic chemistry.   The

quantity of this reagent commonly used per determination (0.25 g) often con-

tains 10-20 ng of arsenic;      the amount varies from portion to portion.

This degree of contamination is of little consequence if the sample  aliquot

used in the determination contains several hundred nanograms of arsenic.  But

if it does not (e.g., in natural waters and small tissue specimens), both the

contamination and the variability of the blank are sources of error.  The

kinetics of the reaction between sodium borohydride and arsenic, are  an addi-

tional complicating factor:  if a sodium borohydride pellet is dropped into an

arsenic-free acid solution, it produces a considerably higher blank  arsenic

reading than if the same pellet is converted to a 1% solution before being added

to the acid solution.  Moreover, this blank response diminishes with the age of

the solution.  The fading of the blank response due to arsenic in the sodium

borohydride appears to result from the gradual adsorption of dissolved arsenic

onto suspended impurities in the reagent solution.  The adsorbed arsenic is

apparently held so tightly that acidification fails to convert it to arsine.

The simplest way to avoid errors from this source is to use the analytic-grade

reagent, which is more expensive, but usually contains less than 0.5 ng of

arsenic per portion (C. Feldman, personal communication).  The efficiency of

sodium borohydride in generating arsine can be impaired by the presence of
                                                                              666a
other substances that react with sodium borohydride.  This effect can be serious.

METHODS OF DETERMINATION OF TOTAL ARSENIC

Molecular-Absorption Spectrophotometry

     Molecular absorption spectrophotometry in aqueous solution has  long been

one of the most reliable methods for determining small quantities of arsenic.
                                 -376-
-------
Because of Its simplicity and low cost, it will probably continue to be widely

used for all but the lowest concentrations.  Arsenomolybdic acid is formed

when arsenate reacts with acidified molybdate.  This heteropoLyacid can be

partially reduced to give a blue color, which develops slowly (^/ 30 min) ,
                                          CQf.
but is stable and free from interferences.     The other color imetric method

in common use involves the bubbling of arsine through a 0.5% solution of the

silver salt of diethyldithiocarbamate in pyridine.  An intense red color is

produced ; absorption is measured at 533 nra.    '

Atomic Absorption

     Atomic absorption (nebulized sample solution plus argon-hydrogen or air-
                                                                        o £ CK
acetylene slot burner) is claimed to give sensitivities of 50-100 ng/ml.

In the flameless atomic-absorption method, a small volume of sample (1-50 pi)

is deposited in a graphite tube or on a tantalum strip.  Strong heating

vaporizes the arsenic and reduces it to As°, which is then determined by atomic

absorption.  The absolute and concentrational detection limits of this method

are good (40 pg and 10 ng/ml, respectively), but care is required in controlling


sample vaporization and in dealing with interferences.     The arsenic can also

be introduced into a gas stream as arsine, with conversion to As  by a flame
                127 ^ftftr*
or a heated tube   '     and detection by atomic absorption.  Detection limits

can be reduced to 1.0 and 0.2 ng, respectively, for these two methods by

accumulating the arsine in a cold trap and releasing it quickly.

Atomic-Emission Spectroscopy

     Arsenic can be determined by atomic -emission spectroscopy with various

types of excitation.  For example, arsine can be accumulated in a cold trap

                                                                            84
and then introduced into a d-c glow discharge in helium (Braman and Foreback

and C. Feldman, personal communication) , giving absolute and concentrational
                                  -377-
-------
detection limits of 0.5 ng and 25 pg/ml, respectively.  Other volatile forms



of arsenic  (e.g., triphenylarsine), introduced into a microwave discharge in



argon,      can give an absolute detection limit of 0.02 ng of arsenic.  An



arsenic-bearing aerosol, introduced into an induction-coupled radiofrequency



plasma, gives a concentration1 detection limit of 40 ng of arsenic per milli-



liter.214a'388b



Neutron-Activation Analysis



     Neutron-activation analysis has the advantages of being nondestructive



(in the many cases in which postirradiation radiochemical separations are not



necessary)  and of being immune from any danger of contamination during post-



irradiation handling.  Its absolute sensitivity is 0.1 ng for a thermal-neutron


          12            2
flux of 10   neutrons/cm -s.   in tissue and mineral samples,  however,  this



sensitivity can seldom be reached.  The activity induced is the 559-keV photo-



peak of arsenic-76.  A relatively great amount of sodium-24 activity is



induced in the sodium present in such samples, and, although the decay of



sodium-24 (half-life,14.96 hr) is faster than that of arsenic-76 (half-life,



26.5 hr), the sodium-24  activity must be allowed to decay for several days



before the arsenic-76 activity can be counted.  This delay does not seriously



interfere with the determination of arsenic at concentrations above a few



parts per million, and the elimination of all chemical treatment of the sample


                                  545a
compensates for the inconvenience.      If greater sensitivity is needed or if



radiochemical interferences appear (e.g., bromine or antimony activities),



chemical-group separations can still be performed to isolate the arsenic-76


  _, ..   315a,547
activity.



Electrochemical Methods



     In the electrochemical methods that have been proposed for determining



traces of arsenic, the arsenic is usually first isolated by volatilization or
                                   -378-
-------
extraction, then converted to the trivalent form and determined polarographi-


      25a
cally.     The most sensitive such technique is differential pulse polaro-



graphy, which has a detection limit of about 0.3 ng of arsenic per milliliter



and can be used in the presence of natural pollutants, such as unfiltered


 ,  ,   517a,549a
sludge.



Gas Chromatography



     Total arsenic can be determined by gas chromatography if the arsenic is



first collected and converted to triphenylarsine.  The collection-conversion



procedure is somewhat long, but the absolute limit of detection is quite low



(20 pg) when an atomic-emission detector is used.



Other Methods



     There are other valid methods of determining traces of arsenic, such as



coulombmetry, x-ray fluorescence, atomic optical fluorescence,     and


                                                699
ordinary and isotope-dilution mass spectrometry.



METHODS OF DETERMINATION OF ARSENIC COMPOUNDS



     Most of the analytic work on separating and identifying arsenic compounds



has been done with substituted arsines and substituted acids of arsenic (e.g.,



methylarsonic and dimethylarsinic).  The compounds have been isolated with paper



chromatography, electrophoresis, volatilization,   and (after silylation   a



or conversion to the corresponding arsine     or iodide   a) gas chromatography.



A specific compound is identified by its retention characteristics, sometimes



in combination with a specific detector for arsenic.  Among the detection


                                       474              84
methods used have been autoradiography,    arc emission,   and microwave



emission.      Absolute sensitivities have been in the picogram range.
                                    -379-
-------
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            Injuries, and Causes  of  Death.  Vol. 1.  Geneva:  World Health


            Organization, 1957.   393 pp.


 787a. Yamashina, H. , Y. Nagae, and S. Sasaki.  Organoarsenic compounds.  Japanese


            Patent 21,072, 1963 to Toa Agricultural Chemical Co.,  Ltd.   (UNVERIFIED)



 790a.   Yeh, S.   Skin cancer in chronic arsenicism.  Hum. Path. 4:469-485,  1973.




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                                 -480-
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                                  TECHNICAL REPORT DATA
                           (1'lcasc read Instructions on the reverse he/ore completing)
]  RS- PORT NO
 EPA-600/1-76-036
a' MILL AND 'JUKI! I L L

 ARSENIC
                                                          3 RECIPIENT'S ACCESSION NO.
             5 REPORT DATE

              NoyembejiJ_2Z6_	
             6 PERFORMING ORGANIZATION CODE
7 AUTHORIS!

 Subcommittee on Arsenic
                                                          8. PERFORMING ORGANIZATION REPORT NO
9 PERFORMING ORGANIZATION NAME AND ADDRESS
 Committee on Medical  and Biologic  Effects of
   Environmental Pollutants
 National Academy of Sciences
 Washington, D.C.
                                                           10 PROGRAM ELEMENT NO.
              1AA601
             11. CONTRACT/GRANT NO.


              68-02-1226
 12. SPONSORING AGENCY NAME AND ADDRESS
 Health Effects Research Laboratory
 Office of Research and Development
 U.S. Environmental Protection  Agency
 Research Triangle Park, N.C. 27711
                                                           13. TYPE OF REPORT AND PERIOD COVERED
              Final
             14. SPONSORING AGENCY CODE

              EPA-ORD
 15. SUPPLEMENTARY NOTES
 16. ABSTRACT
           This report  is  an  in-depth study that attempts to assemble,  organize, and
 interpret present-day  information on arsenic and its compounds,  and  the effects of
 these substances on man,  animals, and plants. Emphasis is given  to the effects of
 arsenic on man, conclusions  are drawn from the evaluation of  current knowledge on the
 subject, and recommendations  are made for further research. Although   arsenic is
 highly toxic in many of its  forms, a number of factors suggest that  it probably is not
 a general pollution problem.   In fact, there are indications  that it may be an essentia
 trace element.  There  is  some evidence that arsenicals can be mutagenic in humans.
 There is epidemiologic evidence that inorganic arsenic is a skin and lung carcinogen
 in man. Skin cancer has occurred in association with exposure to inorganic arsenic
 compounds in a variety of populations, including patients treated with Fowler's
 solution, Taiwanese exposed  to arsenic in artesian well water, workers engaged in the
 manufacture of pesticides, and vintners using arsenic as a pesticide.   Lung cancer has
 been observed to be associated with inhalation exposure to arsenic in  copper smelters,
 workers in pesticide manufacturing plants, Moselle vintners,  and Rhodesian gold
 miners.
         While much arsenic enters the atmosphere from the burning of coal, the
 concentrations are too low to be a matter of concern.
17
                               KEY WORDS AND DOCUMENT ANALYSIS
                  DESCRIPTORS
 Arsenic
 Air Pollution
 Toxicity
 Health
 Ecology
 Carcinogens
b.IDENTIFIERS/OPEN ENDED TERMS  C.  COSATI 1'ieid/Group
                            06 F, H, T
 3 DISTRIBUTION STATEMENT

 RELEASE TO PUBLIC
19 SECURITY CLASS (This Report)

  tlNr.l AS.STFTFH
21. NO OF PAGES

  488
                                             20 SECURITY CLASS (This page)
                                                UNCLASSIFIED
                                                                        22. PRICE
EPA Form 2220-1 (9-73)
                                            481
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