Document ExKJDZ3jLe85wvyMD4XmQdVK0

N ovem ber 19, 1975 SUMMARY OF TOXICOLOGICAL STUDIES ON COMMERCIAL PCB's J. C, Calandra* The urgent need for additional toxicological data becam e ap p aren t when the refinem en t of an aly tical techniques m ade p o ssib le the identification of PCBs in many portions of the ecosystem . To a s s e s s the biological h azard s of these m a te ria ls , chronic feeding studies in ra ts and dogs w ere conducted according to accepted tra d itio n a l p ro c e d u re s w ith A ro c lo rs 1242, 1254 and 1260, It should be pointed out that A ro c lo r 1260 is no lo n g er produced in th is country since it does not m e e t the phy sical sp ecificatio n s fo r its re s tr ic te d u ses (ref, 1), In addition, th ree generation two litte r reproduction and terato lo g y studies in albino ra ts a s w ell as a dom inant le th a l m utagenic study in albino m ice w ere considered to be n ecessary for the broad spectrum safety evaluation of th ese m a te ria ls . Effects such as d ecreases in eggshell thickness described in certain avian species suggested that toxicity and reproduction studies in chickens would be helpful in evaluating possible untoward effects in birds, A lis t of the studies conducted a re p re se n te d in fig u re 1. The ex p erim en tal design of the chronic ra t and dog studies is given in fig u res 2 and 3. It should be noted th at these studies w ere of typical Food and Drug A dm inistration design fo r the evaluation of the safety of d ire c t and in d ire c t food additives. NEV 0 0 8 5 6 9 * In d u stria l B IO -TEST L a b o ra to rie s , Inc. , 1810 F ro n tag e Road, N orthbrook, Illinois 60062, 1 707821 J. C. Caiandra The im portant re su lts in the chronic ra t studies which w ere conducted on C harles R iver ra ts a re sum m arized briefly in fig u res 4, 5 and 6. Food consumption, m ortality and hem atologic, urine and blood chem istry studies did not differ among treatm en t and control groups. The only w eight d e p re ssio n o b serv ed w as a t the 24 m onth point in fem ales fed 100 ppm of A ro c lo r 1254. L iver weight in creases w ere noted in m ales and fem ales fed 100 ppm A ro c lo r 1254 o r A ro c lo r 1250, and in fe m a les only in the 100 ppm group fed A ro c lo r 1242. As expected, the im portant histopathologic changes in the ra t study w ere p re se n t in the liv e rs of the 24 m onth sacrifice anim als and consisted of hepatocellular alterations such as focal hypertrophy, cytoplasm ic lipid changes and in som e an im a ls a t 100 ppm , hepatom as o r cholangiohepatomas. No evidence of hepatocellular carcinogenicity of the A roclors was found in this study. The conclusion that "no evidence of hepatocellular carcin o genicity of the A roclors was found in this study" was reached only after extensive re-evaluation of the original liv er slides as well as additional liv e r sectio n s fro m a ll of the an im a ls a fte r the K im brough re s u lts on A ro c lo r 1260 becam e known to us (ref. 2). NV 0 0 8 5 7 0 2 707822 J. C. Calandra The slides w ere read independently and separately by Dr. Donovan Gordon (ref. 3), P ro fe sso r W ard R ichter (ref. 4), and by P ro fe sso r P, P o u r (ref. 5) of In d u stria l B IO -TEST L a b o ra to rie s , the U n iv ersity of Chicago, and the Eppley Institute for C ancer R esearch, respectively. The in te rp re ta tio n of pathologic changes th a t o ccur in the liv e r as a re su lt of absorption of chlorinated hydrocarbons is a key issu e that needs resolution by the community of pathologists (refs. 6-7). The problem is highlighted in the context of PCBs since th ere a p p e a rs to be d isa g re e m e n t as to w h eth er A ro c lo r 1260 is o r is not a hepatocellular carcinogen (ref. 8). P our re-ev alu ated the K imbrough slides and does not agree with the reported findings. The findings in the chronic dog study a re sum m arized in figure 7. The feeding of A ro c lo r 1260 a t 100 ppm showed an in c re a s e in serum alkaline phosphatase activity and liv er w eights. A slight d e c re a se in body w eight gain w as noted at the 100 ppm dose lev el in m a le s and a t 10 and 100 ppm in fe m a le s . No re m a rk a b le histopathologic changes w ere found. However, evidence of gastrointestinal inflam m atory lesions and u lceratio n s was noted which appear to be sim ila r to those found by Allen in the rh esu s monkey by feeding A ro clo r 1248 (ref. 9). (This m aterial is no longer a product of com m erce (ref. 1).) The design of the dominant lethal m utagenic study is presented in fig u re 8. NEV 0085 71 3 707823 J. C. Calandra No effects re lated to A ro clo r tre a tm e n t w ere seen in the p a ra m ete rs listed in figure 9 - m o rtality , m ating index, num ber of im plantation and reso rp tio n s ite s, num ber of viable em bryos, p r e implantation loss or mutation rates. This finding is in agreem ent with that of G reen et a l. (ref. 10). The standard design fo r a th ree generation two litte r reproduction study in albino ra ts is given in fig u re 10 and the terato lo g y study in figure 11. A b rie f su m m ary of the re s u lts (figure 12) in d icates that none of the A ro d o rs studied produced ad v erse effects in the two litte rs of the fir s t generation. All progeny delivered by fem ale rats in each of 3 generations exposed to d ie ta ry lev els of up to 100 ppm of e ith e r A ro c lo r 1242, 1254 o r 1260 w ere s tru c tu ra lly n o rm a l. A reduction in the m ating index was o bserved in the 2nd and 3rd g en eratio n s of an im a ls fed e ith e r 10 o r 100 ppm. The ab ility of fem ales to_ conceive, c a r r y the d e liv e ry p ro c e ss to p artu ritio n , and to successfully nourish the young was not affected by the 3 A ro clo rs. It should be stre sse d that no changes in the reproductive tra c t of eith er m ale or fem ale ra ts w ere produced by any of the 3 A roclors. Findings in the 3rd generation w ere sim ilar to those in the 1st with no suggestion of any alteratio n s in response as a function of succeeding generations. New 0 0 6 5 7 2 707824 J. C. Calandra Teratologic studies in which albino ra ts w ere exposed to either A ro c lo r 1242, 1254 o r 1260 a t doses of up to 30 m g /k g during rap id organogenesis - gestatio n days 6 through 15 - w ere conducted in our la b o ra to rie s. No evidence of em bryotoxicity as reflected by an in c re ase of fetal resorption or teratogenicity as m easured by com plete external, skeletal, and internal evaluation of fetuses obtained from the A ro clo rtreated fem ales, was obtained. Other experim ental data pertaining to the potential teratogenicity of polychlorinated biphenyls supports the lack of a d v e rse effects. M izunuya (ref. 11) and h is colleagues found no evidence of te ra to g e n ic ity in the r a t when fed a t d ie ta ry lev els of up to 250 ppm. In the m ouse, T oeruek (ref. 12) found th at doses of up to 500 m g/kg w as n o n -terato g en ic although when given on gestatio n days 1 through 6, a decrease in im plantation sites and fetal weights was observed. These doses given on g estation days 7 through 11 fa ile d to produce any changes with re sp ec t to eith er reproductive p a ra m ete rs or teratogenicity. F u rth e r, no evidence of m alform ation was obtained in infant m onkeys d eliv ered to fe m a les fed e ith e r 2. 5, 5 o r 25 ppm A ro c lo r 1248 although the b irth w eights w ere re p o rte d to be reduced (ref. 9). The la st study in this series was a toxicity/reproduction study in white Leghorn chickens (figures 13, 14 and 15). A ro c lo r 1260 a t a ll te s t lev els did not produce a d v e rse effects on the v arious p a ra m e te rs in v estig ated . Egg production in hens fed NEV 008573 5 707825 J. C. Calandra 100 ppm A ro c lo r 1242 o r 1254 was d e c re a se d as w as egg h atch ab ility . In fa c t, poor hatch ab ility of eggs fro m hens fed 8 ppm A ro c lo r 1242 w as found (figure 15). In addition, A ro c lo r 1242 a t 10 and 100 ppm and A ro c lo r 1254 a t 100 ppm w ere a sso c ia te d w ith reduced eg g sh ell th ic k n e ss. Chick v ia b ility w as affected by both su b stan ces a t the 10 ppm dose lev el. Conclusions 1. The n o -e ffe c t lev el fo r the A ro c lo rs in the ch ro n ic r a t and dog studies is about 10 ppm. 2. No teratogenic or m utagenic effects w ere found. 3. No h e p ato c e llu la r c arcin o m a s w e re p re s e n t. 4. The no-effect level in the ra t reproduction study is between 1 and 10 ppm and is the r e s u lt of low m ating in d ices. 5. In the chicken reproduction and teratology stu d ies, effects w ere m o re se v e re w ith A ro c lo r 1242 w ith the n o -e ffe c t le v el being 2 -4 ppm. NEV 0 0 8 5 7 4 6 707826 J. C. Calandra REFERENCES 1 Private communication, Monsanto Company. 2 Induction of L iver Tum ors in Rats by Polychlorinated Biphenyl A ro clo r 1260. K im brough, R. D. et a l. In p r e s s . 3 R eport on H istopathological E valuation of A ro c lo r 1242, 1254 and 1260, M arch 24, 1975, by D. E. Gordon, 4 R eport on H istopathological E valuation of A ro c lo r 1242, 1254 and 1260, M arch 24, 1975, by W ard R. R ichter. 5 R eport on H istopathological R e-evaluation of L ivers for Rats T reated w ith A ro c lo rs , A ugust 1, 1975, by P. P o u r. 6 R eport of a W orkshop on C la ssific a tio n of Specific H ep ato cellu lar Lesions in R ats. Squire, R. A. et al. C ancer R esearch 35:3214 (1975). 7 Subcom m ittee on E nvironm ental C arcin o g en esis of the N ational C ancer A dvisory B oard, N ovem ber 10-11, 1975. 8 R eport on H istopathological R e-evaluation of T issues from F em ale Sherm an Rats Fed A roclor 1260, O ctober 31, 1975, by P. Pour. 9 Response of the Nonhuman P rim ate to Polychlorinated Biphenyl Exposure. A llen, J . R. Fed. P ro c. 34:1675 July (1975). 10 Lack of Dom inant L eth ality in Rats T reated w ith P o ly ch lo rin ated B iphenyls (A roclors 1242 and 1254). G reen , S. et a l. Fd. C osm et. Toxicol. 13:507 (1975). 11 E ffects of PC B s on F e tu s e s and O ffspring in R ats. M izunuya, Y. et al. Shakuhim Eiseigaku Z assh: 15:252 (1975), 12 E ffects of PCB on the Developing M ouse. T oeruek, P . Chemo sphere 2:173 (1973). NEV 0 0 6 5 7 5 707827 Figure 1 TOXICITY STUDIES CONDUCTED WITH AROCLORS 1242,1254 AND 1260 Two-Year Chronic Oral---------------------Albino Rats 9 Two-Year Chronic Oral-------------------- Beagle Dogs Three-Generation Reproduction---------Albino Rats Teratology------------ :----------------------------Albino Rats Dominant Lethal Mutagenic--------------Albino Mice Toxicity / Reproduction----------------------White Leghorn Chickens NfcV 00U 576 Figure 2 TWO-YEAR CHRONIC ORAL TOXICITY STUDY ALBINO RATS AND BEAGLE DOGS Test Material None Aroclor 1242 Aroclor 1254 Aroclor 1260 Dietary Levels (ppm) ---- -- 1, 10, 100 1. 10, 100 1. 10, 100 Number of Animals per Diefary Level Rats MF Dogs MF 50 50 50 50 50 50 50 50 44 44 44 44 NfcV 0 0 8 5 77 Figure 3 TWO-YEAR CHRONIC ORAL TOXICITY STUDY ALBINO RATS AND BEAGLE DOGS Parameters Investigated: Body Weight Food Consumption Hematology Clinical Blood Chemistry (B U N , SAP, SGPT, fasting blood glucose, S C O T -- dogs only) Urinalyses Pathology ( gross, organ weights, m icroscop ic) N tV OObi>7ti Figure 4 INGESTION OF AROCLOR 1242--ALBINO RATS LIVER EFFECTS Increase in Liver Weights -- IOO ppm -- Females Primary Liver Lesions None at 3, 6, 12 month sacrifice 24 month sacrifice--100ppm--Increased incidence of: Nodular Hyperplasia (8/20) Hepatoma (2/20) Cholangiohepatoma (1/20) Hepatocellular Carcinoma i (0/20) NfcV 0U579 Figure 5 INGESTION OF AROCLOR 1254--ALBINO RATS LIVER EFFECTS Increase in Liver Weights -- 100 ppm Primary Liver Lesions None at 3, 6, 12 month sacrifice 24 month sacrifice --100ppm-- Increased incidence of*- Nodular Hyperplasia (13/27) Hepatoma (4/27) Cholangiohepatoma (2/27) Hepatocellular Carcinoma i (0/27) NEV 0 0 8 5 8 0 Figure 6 INGESTION OF AROCLOR 1260-ALBINO RATS LIVER EFFECTS Increase In Liver Weights -- IOOppm Primary Liver Lesions None at 3, 6 , 12 month sacrifice 24 month sacrifice - 1 0 0 ppm-- Increased incidence of: Nodular Hyperplasia (7/27) Hepatoma (5/27) Cholangiohepatoma (2/27) Hepatocellular Carcinoma i ( 0 / 27 ) NfcV t5 tl 707834 j | i !Ii i1 i * i i i fi I j (i Figure 7 EFFECTS OF INGESTION OF AROCLORS -- BEAGLE DOGS Aroclor 1242 ----- no effects Aroclor 1254 -- slight decrease in body weight gain -1 0 0 ppm Aroclor 1260 ----- slight decrease in body weight gain - 100 ppm males -- 10 and 100 ppm females ----- increase in SAP-1 0 0 ppm -- increase in liver weights - 100 ppm i NfcV Figure 8 DOMINANT LETHAL MUTAGENIC STUDY -- ALBINO MICE Test Material Dose* Corn O H--------------------------------------------0 .9 m l / k g Methyl methane sulfonate (M M S )------100 mg / kg Aroclor 1 2 4 2 ------------------------------------ 5 0 0 or 1000 mg / kg Aroclor 1 2 5 4 ------------------------------ 5 0 0 or1000mg / kg Aroclor 1260 -----------------------------------500 or 1000 mg / kg *single dose given i.p. to 12 males / group \ 707835 Nfcv ooat>aj i Figure 9 DOMINANT LETHAL MUTAGENIC STUDY -- ALBINO MICE S No effects related to Aroclor treatment seen in > l Mortality Mating index Number of implantation sites 5 Number of resorption sites Number of viable embryos > Pre - implantation loss Mutation rates 1 707836 NfcV 00d5b<t F ig u r e 10 THREE-GENERATION REPRODUCTION STUDY*-- ALBINO RATS Test Material Dietary Levels (ppm) None Aroclor 1242 I, 10, 1 0 0 Aroclor 1254 I, 10, 1 0 0 Aroclor 1260 I, 10, 100 i *Tw o litters per generation Number of Animals per Dietary Level MF 8 16 8 16 8 16 8 16 ftV OOU5B5 Figure II TERATOLOGY STUDY-- ALBINO RATS Test Material Dose* (m g /kg /d ay) Number of Gravid Rats Corn Oil Aroclor 1242 -- 10 or 30 26 26 Aroclor 1254 10 or 30 26 Aroclor 1260 10 or 3 0 26 i administered on gestation days 6 through 15 (10 doses) NtV OUtittb F ig u r e 1Z EFFECT OF AROCLORS ON REPRODUCTION/TERATOLOGY -ALBINO RATS 1. Reproduction Study First generation -- no effects Second and Third generations - 1 0 and 100 ppm -- decrease in mating Index -- decrease in incidence of pregnancy (Aroclor 1242- no third generation with 100 ppm) 2. Teratology Study -- no effects NfcV 000507 \ ) F i g u r e 13 TOXICITY / REPRODUCTION STUDY WHITE LEGHORN CHICKENS Test Material Dietary Levels (ppm) Number of Animals per Dietary Level MF I None 8 40 Aroclor 1242 1, 2,4, 8, IQ 100 4 20 > Aroclor 1254 1, 10, 100 4 20 Aroclor 1260 l, 0 , 100 4 20 V NEV 0U5 707840 F ig u r e 14 TOXICITY/REPRODUCTION STUDY-- WHITE LEGHORN CHICKENS Parameters investigated body weight food consumption egg production egg quality egg hatchability eggshell thickness chick body weight chick viability pathology i NfcV OOB589 F ig u r e 15 EFFECTS OF AROCLORS ON CHICKEN REPRODUCTION Parameter Dietary Level Causing Effect 1242 1254 Egg Production 100 100 Egg Hatchability 8 100 Shell Thickness 100 100 Chick Viability 10 10 Aroclor 1260-- no effects at any dose level i NEV 0 0 8 5 9 0 ajooKSfGToir voris C apacitor C alt Engineering t ___ .. 2>. McClain :^ S S t P itts b u r g 2-0-U6 -- ............--------------- -- wa -- ........... . . ' -Sr >*>etober 6 , 19T2 oe :6b Bldg. 1T06 - le v Lepartasent - J . C. K irk ee:Qb Bldg. 1502 - B dqtrs. Vorks Bngrg. Bnv. C ontrol - B. A* S m i | Mgr. c c : RiD Center - 601-1256 - B dqtr*. Engrg. Prod. IsprovaB eat - 1 . Boquist c c : Bloomington Vorka - In d u rtria l R elations - A. M axln,-Safety Supvr. I t la one of the functions of th e Headquarters In d u s tria l Hygiene Laboratory to form ulate precautionary statem ents laaued by th e Corporatio n on la b e ls , Base p la te s , d ata sh eets, in stru c tio n manuals, e tc . I t has ease to our a tte n tio n th a t the B aseplate fo r Inerteen cap acito rs has been rec en tly rev ised , end ve have obtained copies of the artw ork fo r th e proposed B aseplates. In our opinion, the precautionary statem ent does not fu lly s e e t the requirem ents of th e Federal Occupational Safety and B ealtb Act and Corpora tio n p o lic y ; and consequently ve are recensealing th a t th e vording be changed to rea d : KAM EHATE THIS EQUIPMENT CONTAINS CELORIHATED IXBR0CARB0HS. EQBEME CARE SHOULD BE TAISJ? TO PREVENT AHI ENTRY 2BT0 THE OT7XROHMENT. PROLONGO SO ? CONTACT ASP PR0LOTGIZ BHEATECNG OF TAPORS SHOULD BE AVOIDED. REFER TO SSTRUCTXON BOOK FOR INFORMATION ABOUT SAFE USE, HANDLING, AND DISPOSAL PROCEDURES. Ve have been unsuccessful In lo c a tin g a copy o f th e In stru c tio n book to vhieb reference Is sa&e on th e B aseplate th a t vas proposed In May, 1972. A pparently, i t Is the fe e lin g of our Sloonington people th a t th is in stru c tio n book vas never a c tu a lly v r itte n nor do they laov vbo, I f anyone, has been assigned to th is p ro je c t. C e rta in ly , eur customers Bust receiv e adequate in stru c tio n s fo r c o rre ct in s ta lla tio n and aaintenanee of e a p a e ito rs. Our In te re s t stems iro n th e f a c t th a t such in stru c tio n s should include precautions fo r safe handling of In erteen and reeo n en d atio n s fo r d isp o sal o f th e contaminated s a te r i a l s . To th is end, ve recom end th a t th e follow ing statem ents be included: / - 906285 B . X>. McClain 2 - ..--October 6, 1972 .......... CAOTIOK ----------- ----- ------ THIS WUHWEHT CONTAINS CHLORINATED HYDROCARBONS. .JR0L0RGn> OR BE&AXED CONTACT OP ZEIS MATERIAL MAT CAUSE SON IRRITATION. BREATHING OP ZEE 1APORS HI fcUPPiClEKT QUANTITIES MAT CAUSE IRRITATION OP ZEE HESPIRATORY TRACT. TAPORS 7ROM TEE HEATED MATERIAL SHOULD HOT SE SHEATHED. -CARE SB5UED BE TAKER TO AVOID SPILLS, USABAS, OR ECOSSSIVZ VAPORIZATION. BSOPSBCE OLOTES SHOULD BE VORR VEST BANDUNG ZEIS PLU3D. PROPERLY PITIED CHEMICAL RESPIRATORS APPROTSD BY ZEE U.S. BUREAU OP XXBES POR PROTECTION AGA2SST ORGAXZC TAPORS SHOULD EE VORN BY ZEDTPISCAIS SOLDERING MINOR IZAIS IP ZEE AVAILABLE VZNTXXATIOR IS BOT ADEQUATE SO REMOTE TAPORS PROM SEE BREATE2HG ZOBE. DISPOSAL ZEIS CELOPJRATED HYDROCARBON PLUS) IS CLASSED AS AH 1S7XR0HMENTAL CONTAMLNANT* CORTAIQtASD PImID SHOULD SE RETURNED TO ZEE SUPRSR POP, DISPOSAL. CORTAMIHATg) PARTS AXD MATERIAL; OTHER TEAM CAPACITOR PLUS, SHOULD EE SHIPPED 2B IZAK-RtOOP CQHTA1HERS TO CEEM-TROL POILCTIOR SERVICES, IR C ., P.O. BOX 200, M3D&. CITY, HEW YORK 11*107 (A RECOiKENDZD DISPOSAL AGENCY AS 0? SRFTBGER 1 , 1972) OR TO ANY ADDICTOKAL VASTS DISPOSAL ASNCY SEAT MAY BE BBC0M4EHSED IB ZEE ^^TTURE You v i l l note th a t ve have need th e te rn "CSL0R3XAZED HYDROCARBONS" re the r then "CELQRBIATED BIPHENYLS". This is se re ly e suggestion. Chlorine ted biphenyls ere one p o u p of eh lo rlo eted hydrocarbons th a t e re c u rre n tly ST sileble fo r use es cep ecito r f lu id s . On th e p o s s ib ility th a t you sig h t a t any tla e d esire to e v itcb to one of these o th er flu id s fo r any of your c a p a c ito rs, th e use of th e broader "CHLORINATED HYDROCARBON" term inology vould not present the n ecessity fo r rev isin g tb s vording on your precautionary ta te a e a ts . I vould appreciate your ecnneats. Very tru ly y o u rs, 906286 j / . E. V ilbur Speieher, A d aln istreto r Beadquarters In d u s tria l Eygieae BBRN003185