Document Xk2xQno5ynRgb91eYo2oKOOd

B 10 - T E S T 1810 FRONTAGE ROAD NORTHBROOK. ILLINOIS 60062 REPORT TO 3M COMPANY 28-DAY ORAL TOXICITY STUDY FC-143 IN ALBINO WCE WITH IBT NO. 8532-10655 inj"A"t B 10 T E ST Ja4uW"1&4Aa REPORT TO 3M COMPANY 28-DAY ORAL TOXICITY STUDY FC-143 IN ALBINO NUCE WITH IBT NO. 8532-10655 I. Introduction A material identifiedas FC-143 (T-1742CoC, Lot No. 269) was obtained from 3M Company for the purpose of conducting a 28-day pilot feeding study in albino mice to establishfeeding levels for long-term studies in that species. This report presents the results of the experiment. J^dAdiAxBi10 -T E S T Idwm4s" Yw- 2 II. Summary A 28-day oral toxicity study was conducted in which groups of albino mice were fed dietary concentrations of 0, 30, 100, 300, 1,000, 3,000, 10,000, or 30,000 parts of FC-143 per million parts of diet. The results obtained during the investigation revealed reduced body weight gains followed by losses in body weight among all test group animals (30, 100, 300 ppm) that were sacrificed after 28 days of testing. An increase in average food consumption was exhibited by all animals fed FC-143. However, excessive wastage of food from their feeders makes interpretation of this parameter difficult. All males and females fed 1,000 ppm or more of FC-143 died within the first9 days of testing. All T-M (300 ppm) mice except 1 male died within 26 days of testing. One animal died in each of the 30 and 100 ppm test groups. No other deaths occurred. Reactions noted during the investigation included roughed fur and muscular weakness for all animals fed 3,000 ppm or greater after the first 4 days of testing. Animals fed 1,000 ppm (T-IV) of FC-143 exhibited similar reactions and signs of cyanosis after 6 days of testing. AU of these reactions were evident in the 300 ppm (T-M) groups aftertest day 9. Some T-Il (100 ppm) animals exhibited slight cyanosis on days 10 and 11 of testing, but appeared normal for the remainder of the study. All 30 ppm (T-I) animals reacted similar to the controls during the 28 days of testing. 8 10 -T E S T lak"a4ve4Yaa 3 Gross pathologic examination of allanimals sacri:Ecedafter 28 days of testingrevealed enlargement and/or discolorationof I or more liver lobules. Statisticaalnalyses revealed significantincreases in the absolute liver weights from males and females of the remaining 30 ppm (T-I), and 100 ppm (T-II)testgroups. Increaseswere also noted in the liverto body weight ratiosfor these animals. Histopathologicexamination of all sacrificedmice revealed treatment-related liver findingsin the males and females of all testgroups examined, 30 ppm (T-I), 100 ppm (T-II),and in two 1,000ppm (T-IV) animalsthatdied during the study. These findings consisted of diffusecytoplasmic enlargement (hypertrophy) of hepatocytesthroughoutthe entireliverlobules (panlobular) accompanied by focalto multifocalcytoplasmiclipidvacuoles. There was also degeneration and/or necrosis of hepatocytes and focal bile duct proliferationamong animals of alltest groups. B 10 T E S T fam 4 The other liver changes observed were those of naturallyoccurring disease or related to the method of sacrifice. Respectfully submitted, INDUSTRIAL BIO-TEST LABORATORIEES, INC. Report prepared by: Bonnie Christop Animal Group Technician Rodent Toxicity Andrew J. M-@rias,B.S. Group Leader Rodent Toxicity Report approved by: &i. Dennis W. Arnold, B.S. Acting Section Head, Toxicology Ffofren-c-e-K-. Ks-@ioshi.@Z,Pl@.D Technical Manager, Toxicology Barrie M. Phillips,Ph.D. Vice President and Director of Research Data Verified by Quality Assurance Unit QiQli=loAistsyAusrs@uarnacnece Supervisor qtc .fodad4ABd 10 -T E S T 5 III.Procedure A totalof 80 young (43 day old) Charles River CD albino mice*, 40 of each sex, was randomly divided into 8 groups of 5 male and 5 female animals each. The animals were allowed an 8 day pre-test period to ad;,ustto the caging system. Groups were fed 0 (Control), 30, 100, 300, 1,000, 3,000, 10,000, or 30,000 parts of FC-143 per millionparts of diet**. AU animals were individually housed with food and water available ad libitum. Daily observations for behavioral reactions or mortalitieswere made. Animals were weighed and food consumption calculationswere made weekly for a 28-day period. At the finalsacrifice,a complete gross pathologic examination was conducted on all surviving mice. Animals that died during the investigationwere also examined grossly. At the time of gross examination, a representative set of organs and other tissues was removed and preserved in neutral buffered formalin for future histopathologicexamination. The weight of the liver of each sacrificedmouse was determined and liver to body weight ratios were calculated. Microscopic examination was conducted on the livers from all sacrificed mice. Also at sacrifice,5 ml of sodium heparinized blood (when possible) and 10 g of whole liver (pooled) from the Control (0 ppm), T-I (30 ppm), and T-II (100 ppm) groups were collectedand quick frozen. The samples were submitted to 3M Company for analyses. Charles River Breeding Laboratories, Wilmington, Mass. Purina Rat Chow, (dry, pulverized diet),Ralston-Purina Company, St. Louis, Missouri. JaA"Aid B 10 -T E S T ld*wA"" Yot4 6 IV. Results A. Body Weights Mean male and female body weight data collected during the investigationare presented in Table 1. All test group animals lost weight. Animals fed 30, 100, or 300 ppm of FC-143 exhibited reductions in body weight gains followed by losses in body weight during the investigation. The body weights were depressed in a dose-related manner. ima@@ B@:1'-0T E S T-IaA"aAW*410= 7 TABLEI TEST MATERIAL: FC-143 28-Day Oral Toxicity Study - Albino Mice Body Weight Data Summary of Mean Values Group and Dietary Level (ppm) Control (0) T-I (30) Sex 0 Body Weight Week: 1 2 3 m 33 34 37 35 F 23 24 25 24 m 33 33 34 32 F 23 24 25 22 4 40 28 32 21** 4-Week Total Weight Change (g/mouse) +7 +5 -1 -2 T-II m 33 28* 25** 23** 25** -8 (100) F 23 21 18** 16** 17** -6 T-II.I m 33 23** 23** 21** 20** -13 (300) F 23 17** 17** 17* - - T-IV m 33 21** - - (1,000) F 23 14** T-V m 33 - - (3,000) F 23 - - T-VI m 33 - - (10,000) F 23 - - T-VII m 33 - - (30,000) F 23 - - Statisticallsyignificantintergroup difference at the 95 percent confidence level . Statisticallysignificantintergroup difference at the 99 percent confidence level. AU animals died. (P < (P < 0.05) 0.01) B 10 -T E ST la&m@ fa4 8 B. Food Consumption Mean weekly food consumption data collectedduring the investigation are presented in Table 11. All surviving test group animals exhibited an increase in the average food consumption when compared to controls. However, in each of the groups fed FC-143, excessive food wastage was observed during the study. JaZahid B 10 -T E S T la4w&tie4Jx4 9 Group and Dietary Level (ppm) Sex Control m (0) F T-I m (30) F T-II m (100) F T-III m (300) F T-IV m (1,000) F T-V m (3,000) F T-VI m (10,000) F T-VTI m (30,000) F TABLE 11 TEST MATERL4,L: FC-143 28-Day Oral Toxicity Study - Albino Mice Food Consumption Data Food Consumption (g/mouse/7 days) Week: 1 2 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 - - - - - - - - - - - - - - - - - - - - - Average Food Consumption (g/mouse/7 days) 68 50 97 73 89 72 122 - - - - = AU animals died. NA= Data not available. Food consumption inadvertently collected as a mean per group for males and females together. B 10 T E S T lakoaam" Y^,e- 10 C. Mortalityand Reactions All animalsin the T-IV (1,000ppm), T-V (3,000ppm), T-VI (10,000 ppm), and T-VU (30,000 ppm) groups died within the first9 days of testing. All mice, but 1 T-M (300 ppm) animal (male No. 31), died within 26 days of testing. One animal died in each of the 30 and 100 ppm test groups during the 28 day feeding period. No other deaths occurred during the study. Reactions noted during the study included roughed fur and muscular weakness for allanimals fed 3,000 ppm or more afterthe first4 days of testing. Animals fed 1,000 ppm (T-IV) of FC-143 exhibitedsimilarreactions and signs of cyanosis after6 days of testing. All of these reactionswere evidentin the 300 ppm (T-M) group after9 days of testing. Some 100 ppm (T-II)animals exhibitedslightsigns of cyanosis on days 10 and 11, but appeared normal for the remainder of the study. All 30 ppm (T-I) animals appeared similarto those of the controls. No other unusual behavioral reactionswere noted. D. PathologicStudies 1. Gross PathologicFindings Gross pathologicexamination of animals sacrificedafter 28 days of testingrevealed enlargement and/or discolorationof 1 or more lobes of the Iiversof all testgroup males and females. No other gross pathologic differencesbetween the testand controlanimals were noted. YaAed4zaBt10 -T E S T loSmaimAs4Ima 2. LiverWeightand Liverto Body WeightRatioData The resultsof the statisticaalnalysesconducted on the absolute liverweights and liverto body weight ratiosare summarized in Table M. Significandtifferencesbetween a testgroup and a controlgroup are designatedby asterisks. Statisticalsliygnificanitncreaseswere noted in the absoluteliverweights among the males and femalesof the remaining test groups (30 and 100 ppm). The increasedliverto body weight ratiosfor these animals can be attributedto the reduced body weights after28 days of feeding. TABLE III MA'lt:HIAI, - @-C-143 28-I)AY ()RAb 'I'LIXICL-1S-lY'tlL)-Y AlitAJ14muick. T*JWAL SACHI@'IC@: ORGAN WL'I(;H'I' Alil) liA'1'11)I)A'I'A -SUMMA14Y (i@' IIK'AN VALUK@S OH(;AN - LilVE'k ------------------------------------------------------------------------------------- I)Ik:'NI'IA CJP(;AN 4LIGHJ' UH(jAN/tiUI)Ywtl(;Iil' PA'I'IU UH(;AN/bH lik:VEII (G) ((;/Ii();i)) (;htltjp t i'Pi4) MAtiES k'bt:-Ai1, ----------------------------------------------------- MAI,i!:S m -----------------m ------- CONTROL o.o 2.128 1.212 5.244H 4 .bot)b 4 ** 4* *1 J. ki3 u** I I . 95 1 u Iti. I u i* loo.o 4 . ti5 8 2.93o** 18.474o* 11.24S/ 300.u 3. . ,,a *44*+*#+ I().,5,,,,a @@4444+** ------------------------------ m --------------------- ----------------------- -s'lA'l16'1 1 A 1,1,1 -4@1,61, 1 @ I A t, S'lA'ii 5'rI I N 'I@N (iHtJ(Pl 1)1r'kt 14@ NC t.-S A'I''I'll 141)l@ (p < .u t) Co14 @ I L)krj(, Lt: A L,I,i @'Ii(piI-ik t Afq1'.1 N'lk'R(,H(IUP i)I k t'tkiP@1,Ct,b A I- I-11 '1ii'l@( P < * 0 1 CLltgF I AVAII,Aht,E, a VALUES FOR 1 REMAINING ANIMAL AFTER 28 DAYS. JaA"AidB 10 T E S T &4wAm" Ina 13 3. HistopathologicFindings Histopathologicexamination was conducted on sectionsof livers taken from allmice sacrificedafter28 days of testingand from 2 T-IV (1,000 ppm) animals thatdied during the investigation.Treatment-related liver findings were noted among the males and females from all test groups examined. These findingsare presented in Table IV. The key for the grading system and abbreviationsused is shown below: + = minimal in severity ++ = mild in severity * & = moderate in severity + @ = marked in severity F = focal M = multifocal D = diffuse The pathologist'sstatementfollowson the next page. 8 10 -T E S T AA"a&%44 Jam IBT No. 8532-10655 3M Company 14 June 9, 1977 I have completed a histopathologicalexamination of H&E stained sections of liverfrom controland test(Groups T-I, T-II,T-III)mice of IBT No. 8532-10655. In addition,sectionsof liverfrom two animals of test group T-IV thatdied during the study were also examined. Treatinent-relatedmorphologic changes were present among both male and female animals of all testgroups. The lesionsconsistedof diffuse cytoplasmicenlargement (hypertrophy) of the hepatocytes situatedthroughout the entireliverlobules (panlobular)accompanied by focalto multifocal cytoplasmiclipidvacuoles of variable size which were random in distribution. There was also degeneration and/or necrosis of hepatocytes and focal bile duct proliferationamong animals of all test groups. The other liverchanges observed were lesionsof naturallyoccurring diseases or relatedto the method of sacrifice. All findingsare tabulated in Table IV. @-X Refnaldo J. Arceo, "M.D. StaffPathologis.t Reviewed and Approved: I Donovan E. Gordon, D.V.M., Ph.D. Diplomate, American College of Veterinary Pathologists Group and Dietary Level (ppm) Animal Number and Sex Control (0) 1-M 2-M 3-M 4-M 5-M 6-F 7-F B-F 9-F 10-F T-1 (30) 12-M 13-M 14-M 15-M 16-F 17-F 18-F 19-F 20-F flepatocellular' flypertropliy F m D ...... ++-+*+ +-++ ...... +4 ...... ++-++#. ...... ...... TABIE IV TEST MATERIAL: FC-143 28-Day Oral ToxicityStudy - Albino Mice IndividuallflotopathologiLciver Changes Hepalocallular Degeneration and/or Necrosis F m D Cytopisomic Vacuoles Lipid F mD Bile Duct Proliferation F m D +-++ + +-++ 4 +-++ + ++ +-++ + +-++ +-++ +-++ ++ +-++ ++ + + +-4+ Lymphoid Infiltrations Periportal F m D + Group and Dietary Level (ppm) Animal Number and Sex T-11 (100) 21-M 22-M 23-M 24-M 25-M 26-F 27-F 28-F 30-F T-111 (300) 31-M T-IV (1,000) 45-M* 50-F* Hepatocellular Hypertrophy F m D ... ...... ...... ... ...... ...... ...... ...... ++-4++ ...... ++-+4-+ Animal found dead. TABLE IV continued TEST MATERIAL: FC-143 28-Day Oral Toxicity Study - Albino Mice IndividualliistopathologiLciver Clianges liepatocellular Degeneration and/or Necrosis F M D ++ ++ ++ ... ++-+++ ...... ++ ++ ++ ++ Cytoplasmic Vacuoles Lipid FM D ++ ++ ++ ++-+4+ ...... ++ +-++ ++-+#+ Bile Duct Proliferation F m D + + +-++ + + + 4 t-41 + ++ ++-+++ Lymphoid Infiltrations Periporial F m D