Document KG7J1Y6mRkg1KXKJjYMq8JwEK

sjoiuijd^ HEPCRT TO THE KOIEWITO CICSUCAL COU7JIY by Oeoll K. Drinker, V..D, Dean and Professor of Physiology Harvard School of Public Health, Boston, tines. September 15, 1930 HONS 048113 i. Experiments to determine the possible toxicity of the following substances: ..................................................................................................... 1. Ghloroosane 8, tjlphonyl phthelete..................................................... 3, Chlorinated diphenyl #1868 .................................... , 4, l^lxture of ohlorlnoted diphenyl and ohlorlnntod diphenyl benzene #5460 .... ............................. , Km 2 3 4 7- II. Some eenorol considerations: 9 1. Erldenoe for the destruction of a mixture of pcnta and bexachlonmphthnlenos (#1006) In the body................ 9 8. ijhe effeot of inoreaelng the sodium chloride In the dlot upon animals receiving toxlo -doses of i'XOOO . . . . 10 3. Tie effeot of high and lov; oaloiun intake on enlmals 1ihallns high concentrations of #1006 ................................. 10 $4. e effect of lnjoctlons of xonthlnn on unlwils haling high concentrations of #1006 ................ 11 M0NS 048124 I. BO'EHII.mfe TO PSITOMB TOB P0S3IB1E TOXICITY 07 THE TOIXOLIMO SUBBTAHCfS 1. CHLORCOSANK On July :i4, 1937, we wore Informed by T, D. Smith tlmt each 6 ounce bottle of 6 soft drink might contain 60 to 70 mgm. of ehlorcosane. Thors was no phannacolOGloi l reason to bellove the ooropound would be Irritating or toxic In any way, and therefore It was deolded to dose rats with it very heavily, the argument bslnj; that If they survlTod heavy dosage without any evidence of damage a verdict of no toxlolty might bo given and much time saved. If, on the other hand, tie large doses proved poisonous It would be necessary to reduce dosage until t non-toxic level was reached. The first of these alternatives proved correct. Experiments. -- August 10, 1937. SO adult white rate ranging In weight from 163 to 311 pa, wore ench fed 70 mom. of ohlorcor.ane in 1 ee, of olive oil dally by stomach tube. These animals showed no evldenoe of damage In any way attributable t> ohlorooaana. They were examined microscopically from the 36th to tbo 92nd dnr. The only oonstant change observed was a possible slight inoroase In th> granularity of the calls of the liver ocrdo, but the pathologist in ohargs fait this to ba within normal llmlta. Tbla finding la doubly roassur ng sines many of the animals developed a chronic suppurative lung condition very prevalent In Boston stock rots at the tiim. But even with this added handicap there was no evidence of damage and we thus conclude that ohlorooanne ev< n In anorrooua doeage Is on lnort material In the body and In oil probability pauses through tbo Intestine unchanged. HONS 048125 a. Dimma phthaiatb On Ansae t 83, 1937, wo woro informed by Dr. R. X. Kelly that diphenyl ' phthalate might be absorbed through the skin or might be Inhaled and swallowed during spray1 ng oparatione. After consultation It was decided to food heavily, and If there eere indications of toxicity to project other experiments. Experiments. -- September 80, 1937. 80 adult white rats ranging In weight from 848 to 9)1 gn. wero each fed 0.5 mfpi. of diphenyl phthalate suspended In 1 oc. of water by stomach tube dally. The animals were from new etook and with one exoeption remained clear of the lung oondltlon which existed Id the ohloroosane croup. During the period of experiment, whloh lasted 92 duys, practically ail the anlmele gained might. Examination of 8 animals killed at Intervale curing this period of test resulted as follows; The organa wore normal crooaly, und on microscopic examination tho llvtr alone fias of possible interest. The changes In the liver varied from those In v.hlob tho oclle of tho llvor cords vicre almost normal, huvlng only a slight swelling and gj-anulerlty of their cytoplasm, to those in whioh these changes wore pronounce 1. In tho lottor there was a moderate degree of vacuolization end a raro hyaLlno body. These alterations It must be understood were the renult of very certain doeogo, elnco tho compound was given by stomach tube and the enlmnln had no possible voy of avoiding it. Neither tho pathologist nor myself wus able to consider than of enough mcmient. to cause us to moke further exporlimnts, I am confident that with the ordinary precautions accompanying apray lacquering no poeniblo ham could be done, even if the concentration of dlpbonyl phthalate In tie lacquor wik fnr nbovo the 5 nor eont figure given us by I>. IS. Kolly HONS 04812b I 8. OHLOItlt'ATZD DITILUIVL -- OOUTOUMD #1860 This material was furnished by the Monsanto Cheraloul Coiqpany. It was said to have a |ohlorlne oontent of 60 per oent, the highest ohlorlna figure of any oompoun^ tested by us during tho past three years. It had boon our 1Repression that; toxicity and chlorine oontent followed one another olosely, but this has not turned out to be the case for #186B.Q!dxlnxanjietlui;Klt)i attsxxBMjsraitdxk Further dlsousolon of this point will follow the description of the experimental observations and their results. Experiment^. -- august 8, 1837. 'Dio compound #1860 was ndminlstored by inhalation, the techniquo being that described in the paper entitled The Problem of Possible Sys ionlo Effects from Certain Chlorinntod Hydrocarbons. Journal of Industrial Hygiene, 1937, 19, 883. For the first observations 80 adult whlto rets were used. They ware exposed os follows: Oroup 1. - 80 animals. Terpersturo to which tho compound v.as houtud to Introduce fume Into air line 140-190" C. Average concentration in air breathed by rats 0,53 men. per cubio meter. Average dally exposure 16 hours. Tho experiment was oontinued for 119 days, eninals being sacrificed at intervals for pathological examination and for tho carbon tetrachloride and alooliol test (see paper previously cited), Tho conditions donuribed above enured swelling end lnci'oure of granularity of tha liver cells. Hyaline lncluaions were rare. These changes wen: quite uniform in nil e:Inala examined after the Slst day, but wore not certainly progressive. Th rats wars very healthy throughout the period and thero was an 'driest uniform c -.In in woight. Tho carbon totrachlori.le-ulcohol teal was pooitlve niter 58 days, v.hioh Indicates that though liver di.muga was apparently alight lomu (lug;1 pa of In"M had boon ilono the organ. Thero use no evidence of HONS 048127 damage In any part of the body except the liver. At the end of 93 days 10 rata, apparently in exoellert condition, wore removed from the experiment and not aside for observation!-, upon liver rocovery. These rocovery animals shoved no cllnioal Chances of any sort. When sacrificed 78 and 141 days after removal from exposure it was observed that the spelling Of the liver jells had disappeared, but the granular and hyalino material remained in t ie liver cells and had apparently beoome porunnent. There was absolutely no progression of damage eftcr removal from exposure. liiese clvmges may be coopered with those produced in rats by inhalation of ohlorlnatcil diphenyl #4463, administered in a similar manner in concontrotiono of 0.57 to 0.93 m^.i. per cable meter over similar poriods of tlms. In the eaoo of this compound the conspicuous difference wuo the for greeter incldonoo of lyalinlzatlon of the liver calls, which in our opinion v;nr the ohornctcrlctlc lesion omipod by #4465 ond mixtures containing it. Ono con therefore conclude that #1260 in low concontretions is definitely loss toxic than #4465. Croup 8. -- After 119 daye, the low concentration of #16D having proved but slightly poisonous, it was docided to increase the concentration in the air breathed by tlie 54 rots then remaining. Temperature end other conditions were msIntel tied ns during the first 119 doys, but by the ueu of vaporisers lnsteeil of 1 ti e iivurtige corcerIrction of i"l?60 in the air vun brought to 6.83 mem. por cubic nntor of air. Exposure to this very high concentration wee continued for 67 days. Dndor these extreme conditions the animals agoin remeinod in perfect haalth. The c irbon tetrachloride t.r.1 uleoV>l test. var. positive but there wen nbr.olutely v.o >lher indleution of 1 Ivor dur-wige ond no evidence of dletin-boroo to other orgur.i. . HONS 048128 -6. When examined at autopsy at the end of 87 deye -- the animals had experienced a toted exposure of 806 dayB -- there were no gross sTldencee of abnormality In any part. On microscopic examination no organ showed changes except the liver. The number of oells containing hyallno and the amount of hyaline In tie cello Involved lncroaaed during thlo period of exposure to the hlghor concentration of #1868. The hyaline was most abundant In the portal zone of the lobuln with very little In the central area. Aftor 48 days of exposure 10 rate rare set aside and were saorifloed 73 days later. They wore In excellent condition when removed and did not change. Grossly, at autopsy, they showed nothing abnormal, and on microscopic examination the liver cells had lost tholr swelling but retalnod Increased granularity and hyaline Inclusions. The experiments on Inhalations of high concentrations of #1860 reenforce the ootiolUBlon that this compound ie of lovi toxicity no compared with #4465 or with chlorlnsVed naphthalenes above trichlornophthalcmo. Tho Question ae to why #1868, the inoo t highly chlorinated compound tested, proved but slightly harmful cannot bo answered with any definitenons. It has been euggosted that the toxicity of alL these chlorinated compounds, even though of varied composition, may depend on the ability of the animal to decompose them eftor lodpmont In the tlssuos, and tiat thin decomposition might be shown by an increase in the chlorine In the urine In suitably conducted feeding experiments. In experiments upon this point we have thovm a definite incroauo in the urinary chlorides v.-hon dogs and rate ware fed e mixture of penta and hexschloruaphthal-on'E (#1006), e compound highly injurious to tho liver and containing C8.6 per cent of chlorine, but similar ohsorvetionr hare not been made with any of tho chlorinated diphenyls or allied compounds. It rnny, however, bo that whan about 65 por cent chlorination Is rase lu-8 tho aubateneoa :'ormnd aro Quito stable in tha body and no ceuno a miui'num of dnnuigo. ' MONS 0481-29 1b oonoljpalon, #1268, if handled with ordinary precautions ao to ventllatlon should bd entirely harmless to workmen. While it cunnot be given an absolutely eliian bill as to health, it is preferable to #4460 and #0460. 4. lapCfURK OF CHLORINATED DIPHENYL AND CHLORINATED DIPHENYL BENZENE -- COMPOUND #0460 This substance was furnished by the Monsanto Chemical Company end v;as said to hays a Ohio:'in* content of 60 per cent, being In this rr.spuot bolow #4460 and far below #12611. Experiments. -- The inhalation tocluilque was used an In the care of jlkt'O and #4465. August B, 1937. 80 odult white rats wore the subjects. Temperature to which the compound wes hoatod to Introduce funo into the air line lCO-lfO0 C. Average concentration in sir breathed by rats 0.085 mgr', per cubic rnnter. Average dally exposure 16 hours. Tlio experiment lasted 119 duys. a certain number of animals wore sacrificed for pathological examination, others wore used for tho oa'bon tetrachloride end alcohol tost, and ntlll others act tcii'o for observation os to recovery from possible damage. In spite o)' tho fact that the concentration in tho rtr breathed l.y tho r-.ir evitrcged ebout h/6 that obtained at comparable timpel::L' > from "HOC', (. mu ' of theso unlmolr became sick end loot weight towards tho end of tho record th of uxposui-o. Wen killed, ouch individuals showed crons mottling of the llvor but no changes In othor organs. On microscoplo oxamlnutlon ewclllng ol' the liver colls, lnu^enncd granularity and hyeline inclusion" ware notl in ' :li killed as early is tho 16th day of exposure. These changes obviously occurred rapidly. Kynllni deposits wore never as nuraoroue us with #lh6D. Animals MONS 048130 removed for recovery after SI and 119 days of exposuro did not gain markedly In weight, and one died for no obvious cause. l.Tion examined at autopsy the swelling of tho liver cells had subsided, but abnormal granularity and hyallnlzation remnlned. Needless tj say tiro carbon tetrachloride and alcohol test was positive vhsnevor used. In vievi of| the faot that f5d60 in euoh low concentration proved so definitely toxip, no higher concentrations wore tested. It seems imperative thnt whenever tils compound Is used in industry, Croat oaro be taken to keop concentrations in the air at on extremely low level. No libertine eon be talien with it, ns with ;"l2Ce. HONS 048131 II. 801S JprjtAl. COHSIDWATIOHS la addition to those tests of cornpounde, certain experiments were done whloh are of interest to those nanufacturinc or using chlorinated hydrocarbons. Details as to| these experiments are of course available but are not included in thlo report since they are not of diroct industrial interests X. E7IDKWC3 FOR TK3 BKS1UUCTI0N OF A MDCTUHK OF PKHTA AND lISXAOIILOniJAKrniAL^^S (#100$) IN TOC BODY It has alleys been a quontion as to whether the chlorinated hydrocarbons which have booh examined by otirsolvoe and by others do hurzu per se or whether toxicity deponnlls on their broalidoun In tho body with tho liberation of somothinG harmful ;o the liver. A partial answer has boon obtained by footling #1006 to rate and docs vhich v;ere on a low chloride diot with uniform excretion of chlorides in the urine. VJion these animals rocoivod the ohloriivsted hyurooarbon (1006) the urinary ohlorldec rose. Tbit; Indicates that the body certainly hus power to detach chlorine from this compound, and it is probtiblo that the same condition in true for allied toxic compounds. It would bo :.:ost interesting to see vdiether ingestion of /'1260 results In similar findings or whethor in tho car.'O of this relatively non-toxic coi^inm*! there ic practically no flitting off of chlorine. IJoithor tine nor out* financial resources permitted such tests* MQNS 048132 -10- S|. TOE EFFECT OF INCREASE.'!; TIE SODIUt: CHEOUltE IN TOE DIET I UPON ANUIAIS RBC3IVIIIG mciO DCSES OF #1006 On the ground thut clilorldes night be fundamentally aaeocioted with toxiolty, e froup of 15 rets was fed a lov.- toxic doso of #1006 and coloured with a similar group on the same dosage of #1006 plus a narked lncreo.no In chloride intake secured by giving 5 oo. per kilogram of body weight of 4 per oent KaCl solution dally. No differences wore found between the two groups, and it may be concluded thct chlorine lncroeso secured through the diet doeo not enhance toxicity. This oxperlno.it was done in order to find out whether increese in chloride Intake during! hot weather might be harmful. A pnrticjilur phaso of the problem, the possible enhancement of typioel skin lesions by increasing. chloride intake, cannot In docldud by experiments on fur-booring animals with no sebuceous glands. All that cod be seld at tho moment Is that increased chloride intake does not increase systemic toxicity. THE EfTUCT OF ai Alii) DO'.i CALCIl'-i liiitJul Cli AMEAlS li.ih'J.E;j iilo;. C.e C'S ,,'lOGu It is noli known that a diet rich in calcium is luirkedly effective li; CO f a provostlug tho|aoute yellow atrophy of tho liver produced by oorbon tetro- ohlorlde (The prevention und Treatment of Curbon Tetrachloride Intoxication. By J>. D. Uxjsor, E.D., a. S. Elliot, Pii.D., end !). II. Hobbiuc, I..G., Journal of the Avcrloon Ledical Ascociotion, 1920, volume 90, pupa 340). To discover whether calclur. in the diet would project L^.-.lnul. liver d:cv. gu -11- from a toxin chlorinated hydrocarbon, 30 adult white rats were plaoed upon a diet of lean borsemeat, eteroh and lord, a combination adequate for maintenance but Very loir in oalolun. Another group was given a diet consisting of dog chow, milk, lettuce and eggs, with added calcium laotete -- a ration very high In calcium. Both groups were oxposed simultaneously to inhalation of high concentra tions of |?1016, an averuge of 11.SI mgm. per cubic meter for 1C hourn a day. After Id days, IS high calcium diet ratB were nllvo and 7 of the low oaloium group. At Intervals animals were killed for examination. In both groups the liver was abnormal grossly and microscopically, and In both groupe animals died from liver damage. It woe impossible to consider that the high calcium diet was in the leust degree protective. It may, therefore, be oor.cluded that adding calcium to the diet of workers either in the form of extra milk or of calcium lactate will not prevont liver damage. THE EFFECT OF INJEC7I0K3 OF XANTHIHE Oil AHL.'ALS ord.halij:o high co;;ci'm/j'iois //ioog In 1937, (r. 0. Neale publiehed a brief paper (The Protective Action of Certain Purineib agalnnt Liver liecrosls Produced by Carbon Tetrachloride and Chloroform. F.ilence, 1937, volume 66, page 63). He clninod that rets lnjeotcd with podium xanthine became markedly resistant to onrbon tetrachloride. This suggested that xanthine might have similar protective power over liver damage from the ohloriimted hydrocarbons on examination in this laboratory. accordingly 36 adult vhlto rats worn cuused to lnhala ("1006 In concentra tions uveroginc 15 mgm, per cubic voter for 16 hours daily. Ono croup of 80 rets wee given BO mer:. of xanthine ouboutanoously evory other day and 40 HONS 048134 of xanthine by etomaoh tube on the alternate days. Hie seoond group of 18 rata had the same exposure to #1006 without xanthine treatment. No differences mere noted and one oannot expeot any effloaoy from xanthine either in the prevention or treatment >f liver disease due to this ohlorlnated hydrooarbon, and in all probability tie earns negative result would be enoountered In connection with allied toxlo uoopoundo. MOWS 048135