Document 91EN7qn9goEmRpmwv85aOVBg6

r*>V. iktfESPCBT TO THZ mSAOTO CEMCAL CODANT by C ec il E. Dr in te r,. 1.D. Pesa and Professor of Physiology E a rre ri School of Public Eoalth, Boston, lisas. r September 15, 1933 C AZb CC3h jc 22 - I . Experiments to determine the poasible to x ic ity of the fo ile d - e s u b sta n c e s: . . . . . . ..................... . .................................. .... 1 . C hlorcosaae .................................................................... . . . . . . 2 . D iphenyl p h t h a l a t e ,......................... . . . . . . . . . . . . . 3 . C hlorinated diphanyl #1269 . . . . . . . . . . . . . 4. liixture of chlorinated diphenyl and chlorinated diphenyl benzene #5460 . . . . . . . . . . . . . . . . . 2 2 3 7. I I . Sosa g e n e ra l c o n s id r tlo n e: . . . . . . . . . . ..................................9 1. Evidence fa r the d estru ctio n of a m ixture of penta and bexachlom aphthalenes(#1006) in th e body . . . . . 9 2 . The e f f e c t o f in c re a s in g th e sodium c h lo rid e in tb s d ie t upon aTvirals re c e iv in g to x ic doses of #1006 . . . . 10 3 . The e f f e c t of hig h and low calcium in ta k e on animals In h a lin g hig h c o n c e n tra tio n s o f #1006 ................. .... 10 4 . The e f f e c t o f in je c tio n s o f x a n th in e on anim als inhai ng hig h c o n ce n tra tio n s o f #1006 . . . . . . . . . 11 S I# TO r^ >TT ?l.rrN"? ZE2 PCSSI3I2 TGCICIT^ m^9 ~ 1 . CHLCHCOSAM2 On J u ly 14, 1937, we Tre inform ed by 7 . D. Sm ith th a t each 6 ounce b o ttle o f a s o f t d rin k might c o n ta in 60 to 70 mga. o f c h lo rc o sa n e . Thera was no pharm acological reaso n to b e lie v e th e compound would be i r r i t a t i n g or to x ic in any way, and th e re fo re i t was decided to dose r a t s w ith i t Tery h eav ily , th e argument being th a t if. they aurriT ed heavy dosage w ithout any evidence of damage a v e rd ic t of no to x ic ity might be given and m c h tim e saved. I f , on th e a o ther hand, th e larg e doses proved poisonous i t would be necessary to reduce dosage u n t i l a n o n -to x ic le v e l was reach ed . The f i r s t o f th e se a lte r n a tiv e s proved co rrect. S xperim ents. -- August 10, 1937 . 20 a d u lt w h ite r a t s ra n g in g in weight from 162 to 210 g s. were each fe d 70 mgs. o f e h lcrco sa n e in 1 c c . of o liv e o i l d a ily by stomach tu b e . These anim als showed no evidence o f damage in any way a t t r ib u ta b le t o ch lcreo ean e. `R.ey were examined m ic ro s c o p ic a lly from the 33t h to th e 92nd day. The only c o n sta n t change observed was a p o s s ib le s lig h t increase in the granularity of the c ells of the liv e r cerds, but the -1X-V:Ta-^^"o"sist*n*`c a r * ^ '? s l 't 't M s ' to'"be w ithin -' > -sir/iv*.j normal lim its . This finding is doubly reassvx'lng s in c e many of th e anim als developed a ch ro n ic suppurative lung co n d itio n very p rev alen t in Boston stock r a t s a t the tim e. But even w ith t h i s addsd handicap th e re was no evidence o f damage and we th u s conclude th a t c h i arco ana even in enormous dosage is an in a r t m a te r ls l in th e body and in a l l p ro b a b ility passes through the in te stin e unchanged. O- ^ 3* j c 2 . DIPEZNYI. rSTHAUCS On * ag u at 23, 1937, we were inform ed by D r. 2 . 2 . f o ll y th a t diphenyl p b th a la te m ight be absorbed thro u g h the ak in o r s ig h t be inhaled and swallowed d u rin g sp ray in g o p e ra tio n * . A fte r c o n s u lta tio n i t was decided to fe e d h e a v ily , and i f th e re were in d icatio n s of to x ic ity to p ro je c t other experim ents. S r o e r i n e r t s . -- September 20, 1937 . 20 a d u lt w hite r a ta ranging in w eight f r o a 242 to 301 gnu were each fe d 0 .5 mgm. o f d iphenyl p b th a la te suspended in 1 c c . o f w ater by stomach tube d a ily . Ris anim als were from new sto ck and w ith one exception remained c le a r of th e lung condition which existed in the chlorcosane group. D uring th e p erio d of experim ent, idilch la s te d 92 days, p r a c tic a lly a l l th s animals gained weight. Scag in atio n of 8 animals k ille d a t lm terralB during th is perio d of t e s t re s u lte d ae follow s: The organa were normal ^ o e s l y , and on m icro sco p ic exam ination th e l l r e r alone was o f p o s s ib le i n t e r e s t . The changes in th e l i v e r v a rie d from those in w hich th s c e l l s of th e H vbt cords were alm ost norm al, haTing only a s l i g h t sw ellin g and g ra n u la rity o f th e ir cytoplasm, to th o se in which these changes were pronounced. In th e l a t t e r th e re was a m oderate de^-ee of v a c u o liz a tio n and~a^rarY h y ^ in e 'b o < ^ T - Theae " a l t r a t ioils' it'm u s t' be understood were th e r e s u l t of v e ry c e r t a i n dosage, sin c e th e compound was given by stomach tube and th e animal* no p o s s ib le way o f av o id in g , i t . N e ith e r th e p a th o lo g is t no r m yself was able to c o n sid e r th e a o f enough masent to cause us to make fu rth er experiments. I am c o n fid e n t th a t w ith the o rd in a ry p re c a u tic n a accompanying sp ray la c q u e rin g no p o s s ib le harm could be done, even i f th e c o n c e n tra tio n of d ip h en y l p b th a la te in th e lacq u er was f a r above the 5 per cent fig u re given us by B. 2 . S a lly . i,K CC3i33 25 f V 3 . CELORIXATZD DIPHTm -- CCL20UO #1253 T his m a te ria l waa fu rn ish e d by tb s Uonsanto Chemical Cccpany. I t *bj s a id to b a rs a c h lo rin e onteat o f 63 p e r c s n t, th a h ig h e st c h lo rin e fig u r e o f any oompouad ta a ta d by us d u rin g th a p a s t th re e y e a r s . I t had been our | inpreasion th a t to x ic ity and chlorine content followed ana another clo sely , b u t t h i s has n o t tu rn e d out to be th a case f a r # l 2fifl.rw*v-!-w-r!r3rir ..i, <>T, m m a n u t n Turther discussion of th is point w ill follow tha description of tha experim ental o b eo n atio n s and th a ir r e s u l t s . E xperim ents. -- au g u st 2, 1937. Tha compound #1263 was adm inistered by in h a la tio n , th e tech n iq u e b ein g th a t d e sc rib e d in th e paper e n t i t l e d The Problem of P o ssib le Systemic S ffa c ts from C ertain C hlorinated Hydrocarbons. Jo u rn al of I n d u s t r i a l E ygiane, 1937, 19. 283. Tor th a f i r s t o b serv atio n s 80 a d u lt w h its r a t s were used. Risy were exposed as fo llo w s: Group 1 . -- 80 an im als. Temperature to which th a compound was heated to in tro d u c e fun in to a i r lin e 140-190 C. Average c o n c e n tra tio n in a i r b re a th ad by r a t s 0 .53 n e a . p e r cubic m eter. Average d a ily exposure 15 hours. The experim ent was co n tin u e d f a r 119 days, anim als b e in g s a c r if ic e d a t in te r v a ls fo r ~Xp~:a+th o lo g ic-a*l! exam ination and--5*f-o:r---t--h--a-*cTaVr-b:*o.-n- t-e---t-rTaTc-h*leoir.rvi,>d*e<-a.nd- -alcr.o-h o l te a t (sea paper previously c ite d ). The c o n d itio n s A s c rib e d above caused sw e llin g and in c re a se of g ra n u la rity o f th a l i v e r c a l l s . H yaline in c lu sio n s were r a r e , ^ .e se changes were q u ite uniform in a l l anim als examined a f te r the 31st day, but w r* not c e rta in ly p ro g r e s s iv e . The r a t s were v e ry h e a lth y throughout th e p e rio d and th e re was an 'alm o st uniform . g a in in w e ig h t. The carbon ta tr a e h i o x id e -a lco h o l t e s t was p o s itiv e a f t e r 52 daya, w h ic h -in d ic ates th a t though liv e r damage was a p p a re n tly s l i g h t soma d a ^ a e of harm had been done the org an . There wes no evidence of Cf4 CC 3 *i 26 damage in any p a r t o f th e body except tb s l i v e r . At th e and of 93 days 10 r a t a , a p p aren tly l a e x c e lle n t c o n d itio n , were removed ir o n th e experim ent aad a et a ald s f o r o b se rv a tio n s upon I I to? r e c r r e r y . These re c o v e ry anim als showed no c lin ic a l-c h a n g e s of any s o r t . tThea s a c r if ic e d 72 and 141 days a f t e r removal iro n exposure i t mas observed th a t the sw e llin g of the liv e r c e lls had disappeared, but the granular aad hyalins m aterial rem ained in th e l i v e r c e l l s and had a p p a re n tly become perm anent. H e re was a b s o lu te ly no p ro g re s sio n of damage a f t e r rem oval from exposure. These changes nay be cornered w ith those produced in ra ts by in h alatio n of c h lo rin a te d diphenyl #4463, ad m in istered in a s im ila r manner in co n cen tra tio n s of 0.57 to 0.93 mga. p e r cubic m eter over s im ila r periods of tim e. In th e case of th is compound th e conspicuous d iffe re n c e was th e f a r g re a te r in c id e n c e of h y a lin in a tlo n of th e l i v e r c e l l s , which in our opinion was th e c h a r a c t e r i s t i c le s io n caused by #4465 and m ix tu res c o n tain in g i t . One can th e r e f o r e conclude th a t #1268 in low c o n c e n tra tio n s i s d e f in ite ly le s s to x ic than #4465. Group 2. -- A fte r 119 days, th e low c o n c e n tra tio n o f #1263 having proved b u t s l i g h t l y p o isono u s, i t was decided to In c re a se th e c o n c e n trn tlsn in th e a i r b re a th e d by th e 54 r a t s th en rem aining. Temperature and o th er conditions were 'm a in ta in e d as d u rin g th e f i r s t 119 days, but by th e use o f 4 v a p o riz e rs In s te a d o f 1 th e average co n cen tratio n of #1268 in the a i r was brought to 6.23 ngn. per cubic m eter o f a i r . Exposure to th is v ery h ig h c o n c e n tra tio n was continued fo r 87 d a y s. Under these extreme conditions the animals again remained la perfect h e a lt h . The carbon te tr a c h lo r id e and alco hol t e a t was pcs i t ir e hut th e re was a b s o lu te ly no o th e r in d ic a tio n of l i v e r damage and no evidence cf d istu rb a n ce to other organa. at* CC3-3' 27  tan examined a t autopsy a t the end of 87 days -- th e animals ex p erien ced a t o t a l exposure of 2C6 days -- th e re were no groea e n d e rc e s of a b n o rm ality l a any p a r t . Ca m icroscopic exam ination no organ showed changes except th e l i f e r . H a number of c e lls co n tain in g h y a lin e and the amount of hyaline la the c e lls la ro lre d increased during th is period of exposure to the h ig h e r c o n c e n tra tio n o f #1268. The hyaline was most abundant in the p o r ta l to n of th e lo b u le w ith r e r y l i t t l e in th e c e n tr a l a r e a . A fte r 42 days of exposure 10 r a t s ware s e t a sid e and were s a c r if ic e d 73 days l a t e r . H ay were in e x ce lle : c o n d itio n when restored and d id not change. G ro ssly , a t autopsy, they shored % n o th in g abnorm al, and on m icroscopic exam ination th e l i r e r c e lls had l o s t t h e i r sw elling but re ta in ed increased gran u larity and hyaline inclusions. The experim ents on in h a la tio n s o f hig h c o n c e n tra tio n s of #1253 re e n fo rc e th e c o n c lu sio n t h a t t h i s compound is o f low to x i c i t y as compared w ith 4465 or w ith c h lo rin a te d n ap h th alan es a b o rt trle h lo r n a p h th a le n e . The q u estio n as to why #1268, th e most h ig h ly c h lo rin a te d compound te s t e d , p ro red but s lig h tly harmful cannot bo answered w ith any d e f tr it e n e s s . I t has been suggested th a t the t o x i c i t y of e l l th e se c h lo rin a te d compounds, a re a though of Yeried compos i t io n , may depend on the a b i l i t y o f th e anim al to decompose th a n a f te r lodgment in th e t i s s u e s , and th a t t h i s decom position might be shown by an in c re ase in the e h lc r l in the u rin e in s u ita b ly conducted feeding experim ents. In experiments upon th i p o in t we hare shown a d e f in ite in c re a se in the u r in a r y c h lo rid e s when dogs and r a t s were fe d a m ix tu re of ponta and h a x ach lcm ap h th alen es (#1006), a compound h ig h ly in ju rio u s to the liT e r and containing 62.6 p er cent of ch lo rin e, but simi o h e e r ra tic n a h are n o t been made w ith any of the c h lo rin a te d diphenyls or a llie d compounds. I t nay, how erer, be th a t when about 63 p e r can t c h lo rin a tio n is reac th e eubatancea formed are q u ite s ta b le In the body and ao cause a - y.mum of damage cc l a conclusion, #1268, I f handled w ith ordinary precautions as to ventiiatio n should be e n tire ly - harm less to workmen. TThile i t cannot he giren an a b s o lu te ly c le a n b i l l as to h e a lth , i t is p re fe ra b le to #4465 and # 5460. 4 . MECTR3 GF CELQSINATSD DIFHBYL AND CHL0S2iAT23 DIPHZIJTT, BENZEOC -- COMPOUND #5460 This su b sta n c e was fu rn ish ed by th e Monsanto Chemical Company and was s a id to h are a c h lo rin e c o n te n t o f 60 p ar c e n t, b ein g in t h i s re s p e c t below #4465 and % f a r below #1268. S r p a r ia e n ts . -- The in h a la tio n tech n iq u e was u sed as in th e case o f #1268 and #4465. August 2, 1937. 80 a d u lt white r a t s were th e s u b je c ts . Temperature to w hich th e compound was h e a te d to Introduce fvae in to th e a i r lin e 140-196* C. Average c o n ce n tra tio n in a ir breathed by r a ts 0.C35 sg a. per cubic m eter. Average d a ily exposure 16 h ours. The 'experim ent la s t e d 119 days, a c e r ta in number o f anim als were s a c rif ic e d fa r p a th o lo g ic a l exam ination, others were used fa r the carbon te tra ch lo rid e and alcohol t e s t , and s t i l l others set aside f o r o b se rv a tio n s a s to reco v ery from p o s sib le damage. In sp ite of th e fa c t th a t the co ncentration in th e a ir breathed by the ra ts averaged about 1/6 th a t obtained a t cosparable tem p eratu res from #1258, a number o f th e s e wt-Miw became sic k and l o s t w eight tow ards the end of the second month o f ex p o su re. TThen Je llie d , such in d iv id u a ls shewed g ro ss s e t t l i n g of th e l i v e r b u t no changes in o th e r o rg an s. Cn m icroscopic exam ination sw elling of th e liv e r c e lls , increased g ran u larity and hyaline in clu sio n s are noted in animals k ille d as e a r ly as the 16th day of exposure. These changes obriously occurred r a p id ly . H yaline d e p o sits were never as numerous as w ith #1268. Animals AC** 29 removed f o r re c o v e ry a f t 51 and 119 days of exposure did s o t g als markedly V in .w eight, and ona died, fo r so obvious cau se. 'Then examined a t autopsy the f e l l i n g of th a liT e r c a lls had subsided, b u t abnormal g ra n u la rity and b y a lin izatio n remained. N eedless to say th e carbon te tr a c h lo r id e and a lc o h o l t e s t v.aa p o s itiv e whenever used. I n view of th e f a c t th a t #5450 in such low c o n c e n tra tio n proved so d e f in ite ly to x ic , no-higher co n cen tratio n s were te s t e d . I t seeas im perative th a t whenever t h i s compound is used in in d u s try , p e a t care be taken to keep ^ c o n c e n tra tio n s in th e a i r a t an extrem ely low l e v e l . No l i b e r t i e s can be taken w ith i t , as w ith #1253. **CP C C 2 ** 3 c 30 C < U . SClg CgTZRAL CONSID-SlAnciS In a d d itio n to th a s s te s t s of compounds, c e r ta in experim ents se ra dons which are of in te re s t to those manufacturing or using chlorinated hydrocarbons. D e tails as to th ese experiments are of course ax ailab le hut are not included in th is rep o rt since they are not' of d irect in d u s tria l in te re s t. 1 . ZV2EENCS ?GR THZ ZESIHUCTXON CT A L2X7JR2 C? PS7IA AND H3AC2L0ENAPEIHA123S (#1006) E l TE2 3EJT I t has alley s been a question as to vhathar the chlorinated hydrocarbons which have been azmined by o u rselv es and by o th e rs do harm p e r as o r w hether t o x i c i t y depends on t h e i r breahdota in the body w ith th e .lib e r a tio n of some th in g h a r ^ u l to th e l i v e r . A p a r t i a l answer has been o b ta in e d by fe e d in g #1006 to r a ts and dogs ttiic h were on a low c h lo rid e d ie t w ith uniform ex cretio n of ch lo rid e s in the u rin e . Then th ese a n tra Is received the c h lo rin a te d hydrocarbon (#1006) th e u rin a ry c h lo rid e s ro s e . This in d ic a te s t h a t th e body c e r ta in l y hiss power to d etach c h lo rin e from t h i s compound, and * -it -ia 'p r o b d tr le " f h a tctiL e'a en B ''ro h d itim '?ls - tr u S '- f iir -ia l i i 8d t o x i c vc'c^cuSSS''.'''''I t'" would be most in te r e s tin g to see A e th e r in g e stio n o f #1263 r e s u lts in s im ile r fin d in g s or w hether i n th e case of t h is r e l a t i v e l y n o n -to x ic compound th e re is p ra c tic a lly no s p littin g o ff of ch lo rin e. N either tim e nor our fin a n c ia l resources perm itted such te s ts . t CC 3 <*3 S 31 2 . TE2 3F73CT C7 INCHZASING 7E2 SCDIZi r ~ ^ UPON ANHIALS RZCZTTZC T dlO DCSLS C? #1006 3^ On t i e ground th a t c h lo rid e s n ig h t he fu n d am en tally a sso ciated w ith t o x i c i t y , a group of 15 r a t s was fe d a l o r to x ic dose of #1006 and ccrp arsd w ith a s im ila r group on the sane dosage of #1006 p lu s a narked increase in ch lo rid e intake secured by giving 5 cc. per k ilo g rsa of body vaight of 4 per c e n t NaCI s o lu tio n d a ily . No d iffe re n c e * e re found between th e two groups, and i t nay be concluded th a t ch lo rin e increase secured through th e d ie t does not enhance to x ic ity . T his experim ent was done in o rd e r to fin d out w hether in c re ase in c h lo rid e in ta k e during hot w eather n ig h t be harm ful. A p a rtic u la r phase of the problem, the p o ssib le enhancecent of ty p ic a l s k in le s io n s by in c re a s in g c h lo rid e in tak e, cannot be decided by experim ents on fu r - b e a r in g anim als w ith no sebaceous g la n d s. A ll th a t can be s a id a t th e moment i s t h a t In c re ase d c h lo rid e in ta k e does n o t in c re a se s y s te s ic to x ic ity . 3 . TSS ZSY2CT 0? El OH AND LCV,' CAL0HZ ZZA IZ ON AIZZALS INHALING HIGH CONCSJSATICIS C? #1006 I t is w e ll ImoTOi th a t a d ie t r i c h in calcium is markedly e ffe c tiv e in p re v e n tin g th e acute yellow atrophy of th e l iv e r produced by carbon t e t r a c h lo rid e (The P re v e n tio n and Treatm ent of Carbon T e tra c h lo rid e In to x ic a tio n . 3y P . I). Larnson, M.D., A. 3 . U in o t, P h .D ., and 3 . H. Hobbins, U .3 ., T cum al o f th e American H edical A sso c ia tio n , 1923, volume 9C, page 342). To d is c o v e r w h eth er calciu m in th e d i e t would proceed ag ainsc l i 7 e r damage AZr r C3*t 32 ( fr o n a to x ic c h lo rin a te d hydrocarbon, 30 a d u lt w hite r a t a ware placed uocn a d ie t o f le an horsam eat, s ta re h and la r d , a com bination adequate fo r maintenance b u t Tery low in c a l c i m . Another ^*oup was given a d ie t c o n s is tin g of deg chow, n i l k , le ttu c e and eggs, w ith added calcium la c ta te -- a ra tio n very high in calcitmu Both groups were exposed sim ultaneously to in h a la tio n of high concentra tio n s o f #1006, an average of 11.21 agm. p e r cubic m eter f o r 16 hours a day. A fte r 16 days, 12 high calcium d ie t r a t s were a liv e and 7 of th e low calcium group. At in te rv a ls animals were k ille d fo r exam ination. In both groups th e l i v e r was abnormal g ro s sly and m ic ro s c o p ic a lly , and in b o th groups anim als d ied from l i v e r damage. I t was im possible to c o n sid e r th a t th e h ig h calcium d ie t was in th e le a s t degree p ro te c tiv e . I t say , th e re fo re , be concluded th a t adding calcium to the d ie t o f workers e ith e r in the form of e x tra m ilk or of calcium la c ta te w ill not prevent liv e r damage. .4 TEZ ZS72CT C7 INJZCTICIE CF XANTHZ3 CN AiJUaiS INEAUSG HIGH CGNCZ:73Ano:S C? #1006 In 1937, R. C. N eale p u b lish e d a b r i e f paper (The P r o te c tiv e A ction of C ertain Purines a g a in st Liver N ecrosis Produced by Carbon T etrach lo rid e and Chloroform S c ie n c e , 1937, volume 86, page 83}. Ha claim ed th a t r a t e in je c te d w ith sodium x a n th in e became markedly r e s i s t a n t to carbon te tr a c h l o r id e . This suggested th a t xanthine might have s im ilar p ro te c tiv e power over liv e r damage from th e c h lo rin a te d hydrocarbons on exam ination in th is la b o ra to ry . a c c o rd in g ly 38 a d u lt w hite r a t s were caused to in h a le #10C5 in c o n cen tra tio n s av erag in g 15 eng*- p e r cubic m eter fo r 15 hours d a ily . Cue ^ c u p of 20 r a t a we* given 20 of x an th in e subcutaneously every o th e r day and 40 ngm. A> CC3441 33 r v C o f x a n th in e by s tc c a c h tub* on tit a a lte r n a t e days. The second group of 18 r a ta had th a sane exposure to #1006 w ithout x an th in e tre a tm e n t. Ko d iffe re n c e s were n o te d and one oannot expect any e ffic a c y ir o n x an th in e e ith e r in th e p re v e n tio n or tre a tm e n t o f liT e r d ise a se due to th is c h lo rin a te d hydrocarbon, and in a l l p r o b a b ility th e same n e g a tiv e r e s u l t would be encountered in connection w ith a llie d toxic ccnpounda. c c 34