Document g2ROMb7X9ozR12RywgV5NQEwQ

. GERRY -ZLEREH BR AR12G-0308~ BOER RS Ww HREEE (21162) OXR (HMMES: 50622) T- 6322.3 Sawfle D-1 FA-1B Chromosome aberndion ded). 1995108238 ARGFRU EHES AT i woos | 005242 - hb STATEMENT FOR ENGLISH TRANSLATION _ . Kashina Laboratory Mitsubishi Chesical Safety Institute Lid." Sponsor: Sumitono 3M Linited Title: Chromosonal Aberration Study of Sasple D-1 in Cultured Mammalian Cells Study No. : 21162 This study vas conducted in Kashina Laboratory of Mitsubishi Chemical Safety Institute. The original report was wriften in Japanese. | hereby declare that this report reflects faithfully the original report as accurately as possible to ay kaowledge. vane NC Vi.llon' TasotsuNishitoni, M.S. Senior Research Scientist we $5201 * 1s fron October 1, 1994, the conpany nase has been changed. 005243 Subsitted to: Sumitomo 3M Linited (REPORT) Chromosoazl Aberration Study of Sample D-1 in Cultured Massalian Cells (Study No. : 21162) Septeaber 30. 1992 MITSUBISHI KASEI INSTITUTE OF TOXICOLOGICAL AND ENVIRONMENTAL SCIENCES 005244 "STATEMENT _ Kashiaa Laboratory Nitsubishi-Kasei Institute of Toxicological and Bavironsental Sciences Sponsor: Sumitomo 3 Limited Title: Chrososomal Aberration Study of Sample D-1 in Cultured Mammalian Cells Study No. : 2162 This study has been conducted in accordance with the GLP Standards applied to Industrial Chesicals of Japan. Managesent: __ Masanobu Katoh sealed Date: Septesber 30. 1992 005245 QUALITY ASSURANCE STATEMENT - kashisa Laboratory Nitsubishi-Kasei Institute of Toxicological and Enviromental Sciences Sponsor Sumitomo 3H Limited Title: Chromosomal Aberration Study of Ssaple D-1 i in CulturedVamal an Cells Study to. : 21162 Study procedures were periodically inspected and the report was audited by Quality Assurance Unit. The standards of laspection adopted were in accordance With the GLP standards applied to Industrial Chemicals of Japan Tonrspaeucdtiiton Study procedure Study resort Dorateaudoift inspection RJoorriill 2130, 11998%2 SeptesJbuelry 303. 1199922 Dtahtee sotfudyredpiorretcintgor lo and to the sanagenent Jhoorriill 2103.. 11990822 SeptesJbuelry 303,. 11996922 Quality Assurance Unit :_ Yoshihiro Miura sealed Date: Septesber 30. 1952 005246 Title: Chrososomal Aberration Study of Sample D-1 in Cultured Mammalian Cells (Study No. 21162) Purpose: To assess the clastogenicity of the test substance by the chrososonal aberration test in cultured sammalian cells Guideline : The Guidelines for Screening Toxicity Testing of Cheaicals of Japan (kaspogyoNo.700, Yakuhatsu No. 1039, 61 Kikyoku No. 1014, 1986) GP: The GLP Standards applied to Industrial Chemicals of Japan (KaspogyoNo.39. Yalahatsu No.229. 59 Kikyoku No.85. 1984) Sponsor + Sumitomo 3H Linited Testing: Kashina Laboratory facility Mitsubishi-Kasei Institute of Toxicological and Bavironsental Sciences 14 Sunayasa, Hasaki-sachi, Kashisa-gun, Ibaraki Study Director + Tamotsu Nishitoni Other Contributors : Akihiko Kido. Miyuki Tanaka Study date : Initiation of the study) (Subsission of the final report) April 10. 1992 Septesber 30, 1992 Unforeseeable circunstance that may have affected on the test results and deviation fron the protocol have not occurred. Retention of records : ALL data, documents. the protocol and the final report will be retained in the safekeeping facility of Kashisa Laboratory for 10 years after the submission of the final report. Further retention will be discussed with the sponsor. 005247 Report of Results of Crromosonal Aberration Test in Cultured Mammalian cells -- 1. General Ttes Nchaegmeicaolf stuhbestannecwe| 2-Fethyl-N-perfruoloaleyl(C-1~8)sulfonylaminolethylacrylate URAC nomenclature) SGtorrucotuutrlailne fooframualnaufoarctruartiinongalsetfhoordm.ula tn case both are unkoom) Aorpdpinearayratenmpceeratuarte wg [ting ine [er~te|e oi n ) chemical TCT [ous] v=8(uain comp1oen1e~nt) : ca. T8K | sofubstthaencneew| Partition w=I~1 c{ao.t2lYoA.Cto0tDa)l) coefficient Purity of the new Solubility soluble in oil chesial sustance [Water|insolubte | - [iotte_| ciomnpucreinztireastion of hydrogui6n0on:e2m0onpom- soluble(z508) seth etTheor 350mm Sforleuonble1.1i.n3 2. Cell line and culture condition Name of cell line|CHL/1U Otained from | GDoi.-,NipLipon Parsacentical Galf serum, 10% 10 Memufacturer (Lob)| GGiibbccoo LLaabb.,_((3SI0PKGTETIISL)) a Te ro -- chrosososes (ode) pCoulture ES [em [v cet woe roe a mse ir 005248 of 59 Gacircle the applicable auaber, and fill in the relevant entries) Ft m ri] (2)Storage Temperature, etc. of $8 (Preparation of $9 (If purchased. fill in spaces to extent possible) [Cm ooie sn ue ra eals w] w[ e || 5.6-benzoflavone(BF) [(b 4h )Ceompomsitiion osf $9 Mi[x me e o1ewm epJwnem[sr e m[n wo]]mr] f[orms T| w+ eomw [aowmwawn ||t_w_]| @treataont condition with S9 Mix (Encircle the applicable number. and fill in the relevant CeT e tem] Amount of $9 E ffE m arriraemeae |m.g =vN v e Ts 1|77 , a (Final concentration) 2/11 005249 4{.TeCesltl GcroonwdtithioInnhainbditipornepTaersattion of the test substance solution - roe Nusber of cells seeded sftorea laNtuumfbaerctoufreprlates for each concentration Witahcotuitvamteitoanbolic Wiatchtisveattaiboonlic Tn] x 100 7 4 x 100 Jl 8Placsntiicn ddiisahseter Becton Dickinson & Co. GPloasitinc ddiisahmeter | Becton Dickinson & Co. 2 plates 2 slates 5 alplate 3 alhlate Preparation SSuhbasttainnce oChroeingcLieennsattlrsautsbisotolanuntcioeofn thoef| Asuabosutanntce of the test 1000 sg/al 1000 mg/ml S00 = s[uSbtsattaenceof(enctihrecletethset ahaiicable one) Dissolved, Others( Gouspended)| Dissolved, | others Couspended > Treatsant of |sAumbosutnasteeosfoleutaicohn test| the cells 0.025 al/plate 0.015 l/plate Nethad of counting of cell usher countin+g heawoictyhLoneter -tixing': with countin+ghe: sowciytthoreter fixing with sta0i1n%ingcr1y0:%swtaifltohrvsiaolliente | staining 10 %wiftohraatine 0.18 crystal violet Remark: + The initiation day of culturing vas defined as 0 dar. 3/11 005250 (@)Cell growth index (Relative value when the value of the solvent-treated group is 100%)_ [CT concentration Cavan Cell growth index () Without metabolic activation 100 x rd [s x[o se o ] T Tmwe w r] ee Without metabolic activation too x | es | fh man [s we [o veo ee Te 1000 x 12 3: The test substance precipitated or floated in culture sediua. wn 005251 5T.TCehsrtorocsoonndailtioAnberarndatipornepTaersattion of the test substance solution _ Withaocuttivasteitoanbolic Witahctmievtaatbioonlic .ell Nusber of cells seeded 4 x 100 fl ww Sfiozrea Manufacturer 6Plcaastiicn ddiisahseter Becton Dickinson & Co. 6Placsntiicn ddiiasmheter | Becton Dickinson & Co. Ncuascbhercoonfcenptlartaetsionfor 2 plates 2 plates [Mofwoesinn t| 5 siiolate 3 alla oCThroeingctieennsattlrsautsbisootlanuntcieoofn tohfe 0 wal . 0 wal Preparation of| sAauobusnttancoef the test of the test Fiese vein] a|wm Solution Sstubastteancofe (tehnecitrecslte| Dissolved. (Guspended)| Dissolved. the applicable one) | Others ) others ) Treataent of | Asauobusnttancoef seoalcuhtitoenst the cells 0.025 al/plate 0.015 al/plate, Tr ee siimthoitbiictor Aaount of $9 Mix | Concentration [__----| os ame Final conc. 0.1ug/al | Final conc. 0.lut/al PT ros n 1h Remark: +The initiation day of culturing was defined as 0 day. 005252 5/11 @eTasbulelt1 2 show the results. - CotearesTo Ga (Judgement of the result Reason for judgement: - The test substance did aot increase the cells with structural chrososonal aberrations aad polyploid cells with or without metabolic activation. These results led to the conclusion that the test substance did not have clastogenic potential. @ Referential matters IL The test substance was insoluble in water and DUSO and was soluble in acetone. But the test substance in NSO was dispersed in culture sediva better than that in acetone. Therefore DSO was selected for solvent of the test substance. 2 In the cell growth inhibition test. SOK inhibition dose of cell growth (TCIDss) was 60ua/al in 24-hour treatuent, T3us/al in 48-hour treatment without metabolic activation and 71 4/af with setsbolic activation. In all treatsent groups. cell srouth index at 1004 8/at or more did not increase or slightly increased and this concentration coincide with that beginaing to precipitate. So it was considered that the test substance wes saturated at 100ug/af or sore (Fig. 1 2). From these results. the highest concentration in chromosomal aberration test was set at 100 g/wd with or without metabolic activation. 3 In chromosomal aberration test, the Lest substance precipitated or floated in culture mediua at 100 ug/at 4 Data were statistically analysed by the x *-test against the negative control Structual aberrant cells anly with gaps were excluded fron this analysis. 005253 6/11 6. Others = Testing facility Kashia Laboratory. Mitsubishi-Kasei Institute of Toxicological and Bavironsental Sciences 14 Sunayama, Hasaki-sachi, Kashina-gun. Ibaraki Tel. 0479(46)2871 Study director Tasotu Nishitoni Senior Research Scientist signed & sealed from 4/10/1992 to 9/30/1992 005254 11 100 a ON g x 10 1 0 10 5 10 S00 1000 200 500 Concentration (x g/nf) @ 24-hour treatment O 48-hour treatment Pig.1 Cell growth index of Sample D-1 (without metabolic activation) 100+ 50 8 z 10 | 0 0 50 100 500 1000 Conce2n0t0r0ation C50u00/) @ With 9 mix (6-hour treatment and 18-hour recovery) Fig.2 Cell growth index of Sample D-1 (with metabolic activation) 005255 8/11 FE| EHrE eEmare -- E re -- e a ---- EEPEEEEy eEprEEeE|e ebf a e ompbEp ebpeee efeefa hae peeepfpo e aay| | ` FFEEE eeaEro fp a e obpa e fo pefa e o efo p| S.$:X Fe C F--oEOgEp bAS E pS p e e fS o pSeo YepSe Y feSYSS |1 i!. gs FEPE EE EeeEEe mE bee]ede ep a fe ppeo fe ee pepe BT eeeaewk [I t Lo PainET m dEoEe fEer Faat Se REerea---- a-- EE PEE EotpREoe]r peo efoeo poeaa e rpfp ee rf pee pp| . BE PeE] fe eeP fpr| E ee e y m . :} EPEEepeEee]afe e a e perfe fe e e per fu | |, = CE[BFEeE ee ]reo e A af ef ae afe es) a 3) + f g. heEEe Ee n ed a Aberrant coliut-g) were statisticully compared with the control by the x3-Geati +4+(p<D.001) ` ii Appendix : List of Control Reagen(s _ [Jo|mowo ie| Negative Control (Vehicle) [ou or Jorma] |oie] Positive Control [sitmenc | we | T19MB | Kyowa Hakko Kogyo Co., Ltd. 1/11 005;258