Document v63RYNMrbOy3oOxQyZ0XE6dMY

9n&uM/iuiL B IO - T E S T JaJxm iotiei,, Jnc. 1810 f r o n t a g e r o a d NORTHBROOK. ILLINOIS 60062 OM HH REPORT TO 3M COMPANY 28-DAY ORAL TOXICITY STUDY WITH FC-143 IN ALBINO MICE IBT NO. 8532-10655 001670  n d ttih ia l B I O - T E 5 T lai& ud& tiM Sy ttc. ) REPORT TO 3M COMPANY 28-DAY ORAL TOXICITY STUDY WITH FC-143 IN ALBINO MICE IBT NO. 8532-10655 I. Introduction A m aterial identified as FC-143 (T-1742CoC, Lot No. 269) was obtained from 3M C om pany fo r th e p u r p o s e of c o n d u c tin g a 2 8 -d a y p ilo t fe e d in g s tu d y in albino m ice to e sta b lish feeding lev els for lo n g -term stu d ie s in th a t s p e c ie s. This rep o rt p resen ts the resu lts of the experim ent. 001671 HdtuUial B I O - T E ST Jaio>iaio*ieSr Sue. 2 ^ II. Summary A 28-day oral toxicity study w as conducted in w hich g roups of albino mice w ere fed d ietary concentrations of 0, 30, 100, 300, 1,000, 3,000, 10,000, o r 30,000 p a rts of FC-143 p e r m illion p a rts of d iet. The resu lts obtained during the investigation revealed reduced body w eight gains followed by losses in body w eight among all te st group anim als (3 0 , 100, 300 p p m ) th a t w e re s a c rific e d a fte r 28 d a y s o f te s tin g . An in crease in av erag e food consum ption w as exhibited by all anim als fed FC-143. H ow ever, excessive w astage of food from th e ir feed ers m akes interpretation of this param eter difficult. All m ales and fem ales fed 1,000 ppm o r m ore of FC-143 d ied w ithin the fir s t 9 d ay s of te stin g . All T -m (300 ppm ) mice except 1 m ale died w ithin 26 d a y s of te s tin g . O ne an im al d ie d in e a c h of th e 30 a n d 100 ppm te s t g ro u p s . No o th e r d e a th s o c c u rre d . Reactions noted d uring the investigation included roughed fu r and m uscular w eakness for all anim als fed 3,000 ppm o r g re a te r after the firs t 4 d ay s of te stin g . Animals fed 1,000 ppm (T-IV ) of FC-143 exhibited sim ilar reactio n s and sig n s of cyanosis after 6 days of te s tin g . All of th ese re a ctio n s w ere e v id en t in th e 300 ppm (T -H I) g ro u p s after te s t d ay 9. Some T -II (100 ppm ) a n im als e x h ib ite d s lig h t c y a n o s is on d a y s 10 a n d 11 of te s t in g , b u t a p p e a re d n o rm a l fo r th e re m a in d e r of th e s tu d y . A ll 30 ppm (T -I) a n im a ls r e a c te d sim ila r to th e c o n tro ls d u rin g th e 28 d a y s of te s tin g . 001672 HtuLubUal B I O - T E S T Jaiotalvuei, oe. 3 G ro s s p a th o lo g ie ex am in atio n of a ll a n im a ls s a c rific e d a f te r 28 d a y s of testing revealed enlargem ent an d /o r discoloration of 1 o r m ore liver lobules. Statistical analyses revealed significant increases in the absolute liver w e ig h ts from m ales an d fem ales of th e re m a in in g 30 ppm ( T - I ) , an d 100 ppm (T -II) te s t g ro u p s . In c re a s e s w e re a lso n o ted in th e liv e r to body w eight ratios for these anim als. H istopathologic exam ination of all sacrificed mice re v ealed treatm en t-related l i v e r fin d in g s in th e m ales a n d fem ales of a ll te s t g ro u p s e x a m in e d , 30 ppm (T -I) , 100 ppm ( T - I I ) , an d in two 1,000 ppm (T -IV ) a n im a ls th a t d ie d d u r in g the study. T hese findings consisted of diffuse cytoplasm ic enlargem ent (hypertrophy) of hepatocytes throughout the entire liv er lobules (panlobular) accom panied b y focal to m ultifocal cytoplasm ic lip id v a c u o le s. T h e re w as also degeneration an d /o r necrosis of hepatocytes and focal bile duct proliferation among anim als of all test groups. G01673 y n d u ih ia l B I O - T E S T 2 a U *d to *isi, Su c. 4 The other liv er changes observed w ere those of natu rally o ccu rrin g d isea se o r re la te d to the m ethod of sa c rific e . Respectfully subm itted, INDUSTRIAL BIO -TEST LABORATORIES, INC. Report prepared by: _____________ Bonnie C hristo p h er Animal Group T echnician Rodent Toxicity Andrew J . M arias, B .S . Group Leader Rodent Toxicity Report approved by: 0J- D en n is W. A rn o ld , B .S . Acting Section Head, Toxicology -uZt F lorence K. Kjfioshibi', P h .D . Technical M anager, Toxicology B arrie M. P h illip s, P h.D . Vice P resid e n t and D irector of R esearch Data V erified by Q uality A ssurance Uni. U Quality A ssurance S upervisor qtc 001674 H*uf*ubiial B I O - T E S T Jabo*aio*i*d* 9"c. 5 HI. P ro ced u re A to ta l of 80 young (43 d a y o ld ) C h a rle s R iv e r CD a lb in o m ice*, 40 of each s e x , was random ly divided into 8 g ro u p s of 5 male and 5 female anim als each . The anim als w ere allow ed an 8 day p re -te s t p erio d to a d ju st to the caging system . G roups w ere fed 0 (C ontrol), 30, 100, 300, 1,000, 3,000, 10,000, o r 30,000 p a rts of FC-143 p e r m illion p a rts of diet**. All anim als w ere in d iv id u ally housed w ith food and w ater available ad lib itu m . Daily observations for behavioral reactions o r m ortalities w ere m ade. Animals w ere w eighed and food consum ption calculations w ere m ade w eekly for a 28-day period. At the final sacrifice, a complete gross pathologic exam ination w as conducted on all su rv iv in g m ice. Animals th at died d u rin g the in v estig atio n w ere also exam ined g ro ssly . At the time of g ro ss exam ination, a re p re se n ta tiv e set of organs and other tissues was rem oved and p reserv ed in neu tral buffered form alin for future histopathologic exam ination. The w eight of the liv er of each sa c rific e d m ouse w as d eterm in e d an d liv e r to body w e ig h t ra tio s w e re calculated. M icroscopic exam ination w as conducted on the liv e rs from all sacrificed m ice. Also at sacrifice, 5 ml of sodium hep arin ized blood (w hen p o ssib le) and 10 g of w hole liv e r (p o o le d ) from th e C o n tro l (0 p p m ) , T -I (30 p p m ) , a n d T -II (100 ppm ) g roups w ere collected and quick fro zen . T he sam ples w ere s u b m itte d to 3M C om pany fo r a n a ly s e s . * C harles R iver B reeding L ab o rato ries, W ilmington, M ass. ** P u rin a R at C how , ( d r y , p u lv e riz e d d ie t) , R a lsto n -P u rin a C o m p an y , S t. Louis, M issouri. 001675 S tu L a in ia l B I O - T E S T JiaiM afo titi, Hhc. 6 > IV. R esults A . Body W eights Mean male and female body w eight data collected d u rin g the investigation are p re sen te d in T able I. All te st group anim als lo st w eight. A nim als fed 30, 100, o r 300 ppm of FC-143 ex h ib ite d re d u c tio n s in body w eight gains followed by losses in body w eight d u rin g the in v estig atio n . The body w eights w ere d epressed in a dose-related m anner. 001676 SitatiAhtal B I O - T E S T Jai&iato'iUy Sue. 7 ) TABLE I TEST MATERIAL: FC-143 28-Day O ral T oxicity Study - A lbino Mice Body W eight Data Summ ary of Mean Values G ro u p and D ietary Level (ppm ) C o n tro l (0) T-I (30) T-H (100) T-IH (300) T-IV (1.000) T-V (3,000) T -V I (10,000) T-VII (30,000) Sex 0 M 33 F 23 M 33 F 23 M 33 F 23 M 33 F 23 M 33 F 23 M 33 F 23 M 33 F 23 M 33 F 23 Body W eight Week: 12 3 34 37 35 24 25 24 33 34 32 24 25 22 28* 25** 23** 21 18** 16** 23** 17** 23** . 21** 17** 17* 21** 14** - - - - -- - -- - - -- ---- 4 40 28 32 2i** 25** 17** 20** - - - - - 4-W eek Total W eight Change (g/m ouse) +7 +5 -1 -2 -8 -6 -13 - - - - * - * S ta tis tic a lly s ig n ific a n t in te r g r o u p d iffe re n c e at the 95 p e r c e n t (P < 0 .0 5 ) confidence le v e l. ** S ta tis tic a lly s ig n ific a n t in te r g r o u p d iffe re n c e at th e 99 p e r c e n t (P < 0 .0 1 ) confidence le v e l. - = All anim als died . 001677 f ad u itn ia l B I O - T E S T Jia io 'ia io 'U e i, Hkc. 8 ) B . Food Consumption Mean w eekly food consum ption data collected d u rin g the investigation a re p re sen te d in T able II. All su rv iv in g te st group anim als ex h ib ited an in c re ase in the av erag e food consum ption w hen com pared to c o n tro ls. H ow ever, in each of the groups fed FC -143, excessive food w astage was observed during the study. 001678 ittdudiUal B I O - T E S T JaiMafatte, 9tie. 9 TABLE H TEST MATERIAL: FC-143 28-Day O ral T oxicity Study - Albino Mice Food Consum ption Data G ro u p and D ietary Level (ppm ) C o n tro l (0) T -I (30) T-II (100) T -in (300) T -rv (1,000) T-V (3,000) T-VI (10,000) T-VH (30,000) Sex M F M F M F M F M F M F M F M F Food Consum ption (g/m ouse/7 days) Week: 12 3 4 NA 40 110 54 NA 42 52 57 NA 84 118 89 NA 87 57 76 NA 56 136 76 NA 63 77 77 NA 121 143 101 NA 133 96 -- NA NA -- " -- - - -- ----" A verage Food Consum ption (g/m ouse/7 days) 68 50 97 73 89 72 122 -- - - - - -- - ----" -- - ** " - - = All anim als died. NA= Data not available. Food consum ption in advertently collected as a mean p e r group for males and fem ales to g eth er. ) 001679 SttiLti&Ual B I O - T E S T jlai&iafMiMi Stte. 10 C . M ortality and Reactions All anim als in the T-IV (1,000 p p m ), T-V (3,000 p p m ), T-V I (10,000 p p m ), and T-VII (30,000 ppm ) groups died w ithin the firs t 9 days of te stin g . All m ice, b u t 1 T-HI (300 ppm ) anim al (male No. 31), died w ithin 26 d a y s of te s tin g . O ne anim al d ie d in each of th e 30 a n d 100 ppm te s t g ro u p s d u rin g th e 28 d a y fe e d in g p e r io d . No o th e r d e a th s o c c u r r e d d u r in g the study. Reactions noted during the study included roughed fu r and m uscular w eakness for all anim als fed 3,000 ppm or m ore after the firs t 4 days of te stin g . Animals fed 1,000 ppm (T-IV ) of FC-143 exhibited sim ilar reactio n s and sig n s of cyanosis after 6 days of te stin g . All of these reactio n s w ere e v id e n t in th e 300 ppm (T -H I) g ro u p a fte r 9 d a y s of te s tin g . Som e 100 ppm (T -H ) anim als e x h ib ite d s lig h t s ig n s of c y a n o sis on d a y s 10 a n d 11, b u t a p p e a re d n o rm a l fo r th e re m a in d e r of th e s tu d y . A ll 30 ppm (T -I) a n im a ls a p p e a re d sim ilar to those of th e c o n tro ls. No o th er u n u su a l b eh av io ral reactions w ere noted. D . Pathologic Studies 1. G ross Pathologic Findings G ro ss p a th o lo g ic e x am in atio n of a n im als s a c rif ic e d a fte r 28 d a y s of testing revealed enlargem ent an d /o r discoloration of 1 or m ore lobes of th e liv e rs of aU te st g ro u p m ales an d fe m a les. No o th er g ro ss pathologic differences betw een the test and control anim als w ere noted. G016S0 SneLukial B I O - T E S T a&Mafotiei, Hue. 11 2. L iv er W eight and L iv er to Body W eight Ratio D ata The resu lts of the statistical analyses conducted on the absolute liv e r w eights and liv e r to body w eig h t ra tio s a re sum m arized in T ab le in . Significant differences betw een a test group and a control group are designated by a ste risk s. Statistically significant increases w ere noted in the absolute liv er w eights among the m ales and fem ales of the rem aining test g ro u p s (30 an d 100 p p m ). T h e in c re a se d liv e r to b o d y w eig h t ra tio s fo r th e s e an im als c an b e a ttr ib u te d to th e re d u c e d b o d y w e ig h ts a fte r 28 d a y s of feeding. 001681 BIO-TEST TABLE I I I TFST MAJFKIAL - FC-143 2 8 - D A Y URAL TUA 1C I T Y S i U U Y - A1.HJ.NU M1CF FINAL SACH1F1CF ORGAN WFI GHT AND R A T I O OATA SUMMARY UK MKAN VAL UF S ORGAN - Li Vf cR GhOlJH DI F i AHY LFVFI, (RRm) ORGAN WFIGHT (GJ MALFS FFMALlt-.S KGAN/HODY WFIGHT RATIO ( G / 100 G) MALFS Fb.MA L F S RGAN/bKAIN WFIGHT RATIO CG/G) MALFS FFMALFS CONTROL T-J 1- I l 1-1 I I U.O 30. U 100.0 300. U 2.12 5.70 7** 4.058** 3 . 2 30a 1.272 3 . 8 3 b** 2. y 3D** 44444444 5.2448 17.9530 18.4740* 1b . 1b<ula 4 . bOOb 1 8 . 1 /U 3* 17.245/ 44444444 *4444444 44444444 44444444 44444444 44444444 44444444 4444444* S T A I 1ST 1C A L I , Y S I G N I F I C A NT IN' F h GHu UP D l F F t R b . N C F S AT T Hb. I R C . 0 5 ) C U Nb 1DFNCb L b i Vb L . * * S T A T I S T I C A L L Y S I G N I F I C A N T 1 iVl FKGHUUP Dl b b c HFUCb.S Al l i t F W S , I R C . 0 1 } C ONF 1Db nC b L t V F L . DATA nUT A V A l L A H L t * VALUES FOR 1 REMAINING ANIMAL AFTER 28 DAYS. O K <n 00 fO i D tvfiu U ia l. B I O - T E S T Ja io ta fo tto i, Hue. 13 \ 3. H istopathologic Findings H istopathologic exam ination was conducted on sections of liv e rs ta k e n from all m ice s a c rific e d a fte r 28 d a y s of te s tin g a n d from 2 T -IV (1,000 ppm ) anim als that died d u rin g the investigation. T reatm ent-related liv e r findings w ere noted among the m ales and fem ales from all test groups exam ined. T hese findings are presen ted in Table IV. The key for the grad in g system and abbreviations u sed is show n below: + = minimal in sev erity ++ = m ild in s e v e rity +++ = m o d e ra te in s e v e r ity ++++ = m a rk e d in s e v e rity F = focal M = multifocal D = diffuse T he path o lo g ist's statem ent follows on the n ex t p ag e. G016S3 H H itudhial B I O - T E ST Ja k u d v tls L, Une. EBT N o. 8532-10655 3M C om pany 14 J u n e 9, 1977 I h a v e com p leted a h isto p a th o lo g ic a l ex am in atio n of H&E s ta in e d s e c tio n s of liv e r from control and test (G roups T -I, T -II, T-IH ) mice of IBT No. 8532-10655. In addition, sections of liv e r from two anim als of te st group T-IV th at died during the study w ere also exam ined. T reatm en t-related m orphologic changes w ere p re se n t among both male and female anim als of all test g ro u p s. The lesions consisted of diffuse cytoplasm ic enlargem ent (hypertrophy) of the hepatocytes situated throughout the e n tire liv e r lobules (p an lo b u lar) accom panied b y focal to m ultifocal cytoplasm ic lipid vacuoles of variable size w hich w ere random in d istrib u tio n . T here was also degeneration an d /o r necrosis of hepatocytes and focal bile duct proliferation among anim als of all test g ro u p s. The other liv er changes observed w ere lesions of naturally occurring d ise a se s o r re la te d to th e m ethod of s a c rific e . All fin d in g s a re tab u lated in T able IV. Reviewed and A pproved: Staff Pathologist Donovan E. Gordon, D.V.M., Ph.D. D iplom ate, A m erican College of V eterinary Pathologists C016S4 Group and D ietary Level (ppm) Animal Number and Sex Control (0) 1-M 2-M 3456789- F 10-F 12-M 13-M 14-M 15-M 16-F 17-F 18-F 19-F 20-F Hepatocellular ' H ypertrophy FM D M M M F F F 44-444 4 4 -+ 4 4 444 44~444 44 44"444 44-444 44-444 44-444 TABLE IV TEST MATERIAL: FC-143 28-Day O ral T oxicity Study - Albino Mice Individual Histopathologic Liver Changes H epatocellular D egeneration and/or Necrosis F MD Cytoplasmic Vacuoles - Lipid F MD Bile Duct Proliferation F MD 4-44 4-44 4 4-44 4-44 44 44 44 44 4-44 4 - 4+ 4-44 4 - 4+ 4-44 4 4-44 44 4 4 4 4 Lymphoid Infiltrations Periportal F MD C ytoplasm ic Vacuoles H ydrophlc F MD O m tn + 001.655 in Group and D ietary Level (PP" ) Animal Number and Sex T-II (100) 21-M 22-M 23-M 24-M 25-M 26-F 27-F 28-F 30-F T-Ul (300) 31-M T-IV (1,000) 45-M* 50-F* H epatocellular H y p ertro p h y FM D ++-+++ H ++-+++ *f-+++ +++ ++-+++ 4-*-+++ * Animal found dead TABLE IV continued TEST MATERIAL: FC-143 28-Day O ral T oxicity Study - Albino Mice Individual Histopathologic Liver Changes H epatocellular D egeneration and/or Necrosis F MD ++ ++ ++ Cytoplasmic Vacuoles - Lipid FM D +-++ -M- ++ ++-+++ +-++ Bile Duct Proliferation F MD + + + ++-+++ + Lymphoid Infiltrations Periportal F MD C ytoplasm ic Vacuoles Hydrophlc F MD + + +-++