Document 9Jq8QoJx7z7qOXn5YL59Z36p6

AR226-2894 COMPLETE REPORT IN MB FOLDER, FOR DU FONT USE ONLY Copies co E. I du Pone de Nemours and Company Baskell Laboratory for Toxicology and Industrial Medicine Elkcon Road, Newark, Delaware l9?ll HASKELL LABORATORY REPORT MO. 88-81 Macer-ial Tested Haskell No. 13.383 Stud'- Initiaced/Cgffipleted .1/5/80-5/30/80 Material Subciffed bv aemicals & Pigments Department Jackson Laboratory __ SUBACUTE INHALATION TOXICITY STUDY OF< ION E(ATS III) Introdugtion; Tne purpose of this study was to observe the effects on assle rata of repeated inhalation exposureeo IB--------IH vapors. In a previous study fl^MUlMfr the LC30 for ^|^------1 was decenoined co be 1G7 mg/1. Design conecncratioas for this study are 0 ng/1 (Group I), 1.0 mg/1 (Group II), and 10.0 mg/1 (Group o(fl------bin air. Procedure! A. Animals Male CtliCW rats were housed in pairs in stainless steel 'dice mesh csgea. Purlna Certified Rodent Chow i'5002 and water were available aa libieun. All rats were held under these conditions for a 1 week pre-test period ee insure suitability. B. Exposure Protocol Groups of 10 rats weighing 234.0 to' 245.0 grams were exposed to II atmospheres containing fUB------9 (either 1.0 mg/1 - Group or 10.0 mg/1 III) Group 6 hours/day, 5 davs/week for 2 weeks. Control rats (Group I) weighing ^34.0 ro 242.0 grams -/ere siBultaneously exposed to air only. All rats were weighed and observed daily (except weekends and holidays) throughout the exposure period and a 14-day recovery period. Company Sanitized. Does not contain T.SCA cm C. Generation Acaospheres of the ceat material were generated by syringe driving the compound through a Spraying Systems nebulizer. The syringe and nebulizer were heated to a remperature of approxiaately 7QC. The aerosolized material sprayed onto the heated (50"C) surface of a 3-neck round bottom flask. Housellne air swept che evaporated material into 20 liter glass exposure chambers. D. Analytical Chamber samples were collected at approximately 30-minute intervals. A known volume of chamber atmosphere was drawn through 2 candem midget impingera containing FC-113 as a trapping solvent. These samples were analyzed using a Hewlett Packard Model 5710 gas chromatograph equipped with a flame ionization detector. The column used was a 6' x 1/4" O.D. glass coil packed with 20X S.E. 30 on 80/100 mesh ChromosorbS1 W-HP. The G.C. oven temperature was programmed with an initial 2 minute hold at 50C, increased at 32"/minute to 200 "C, and held there for 2 minutes. G.C. analysis was based on tL. Ukomponect. This peak was identified on the basis of che GC* peak height area and the reported sample composition. Sample concentrations were determined by comparison wi-h standards prepared by dilution of the test compound. Chamber concentrations were expressed as the equivalent ^Jf----concentration (mg/1). Oxygen and chamber temperature were aonicored throughout- each exposure. E. Clinical Measurements Clinical laboratory measurements were made on urine samples collected overnight following ehe 9th exposure and the 13th day of recovery. Blood saaplas were taken from the rats' tails afcar the 10th exposure and the l^ch day of recovery. Detailed clinical chemistry procedures and indices are outlined in the Clinical Chemistry report (1). P. Pathology After the 10th exposure, 5 rats from each group were selected at random and sacrificed for gross ind histopathologiral examination. Remaining rats were sacrificed on the 14ch day of recovery for an identical examination. Pathological procedures aaa indices are included in the Pathology reoort (2). G. Organ and Body Weight Analysis Organ weights and organ-co-body weight ratios were determined for che II). heart, liver, lungs, kidnevs. Spleen, testes, and thvrois (Appendices I and Company Sanitized. Does not contain TSCA cm Result During exposures, low boiling componencs evaporated readily while high bcx-ing solids plugged the generation apparatus. This problem accounts for the high standard deviation and range for chese exposure 'oncer.c rations. Detailed data is listed in following Tables. GROUP II design Level - 1.0 ng/1 Exposure No. 1 2 3 4 5 6 7 8 9 10 Average (me/I) 2.02 1.77 0.99 1.^ 1.91 1.1! 3.58 0.97 0.; 0.75 S.D. 2.56 2.10 0.90 2.39 2.05 1.67 3.17 0.57 0.70 0.89 Range 0.57-7.36 0.43-7.24 0.23-2."4 0.14-8.09 :.L4-6.23 0.16-5.31 0.08-8.26 0.27-2.25 0.14-2.17 0.0-:.68 GROUP III Design Level - 10.0 ng/1 Exposure No. 1 2 3 4 5 6 7 8 9 10 Average (ag/1) 14.6 6.7 9.4 7.8 7.3 8.9 11.0 12.3 8.0 9.8 S.D. 12.3 5.1 5.2 5.2 1.6 4.1 5.0 10.1 4.3 2.8 Range 3.7-48.50 1.8-16.0 2.3-17.3 2.9-19.8 4.6-9.8 4.8-16.1 4.0-21.4 4.6-40.6 2.2-16.2 5.7-17.3 The overall averages were 15 j^ 1.9 (Group II) and 9.6 ^ Chamber temperature yss jC28C during all exposures. ^20X during all exposures. 6.4 (Group III). Chaimber oxygen was Company Sanitized. Does not contain TSCA cm i.-K-- T iwifpiiyf Kr-f" v--'<i in'i!)'"^"^ Clinical Observations; During exposure, Group IT rats exhibited a reduced response to sound. Group ITT rats showed a reduced response to sound and rapid breathing. Rapid breathing, however, was observed only .-ifter high concentration excursions. III After one exposure, olid weight loss was observed in Group I and Group II rats followed by normal rate of weight gain. Following 2 exposures. Group \ rars exhibited moderate to severe weight loss followed by recovery of normal rate of weight galn III One Group rat was found dead prior to the second exposure. Group TIT rar died during the second exposure. Another Clinical Cheats fry: Ho effects were seen in Group II rats after exposure 10 III or 14 days recovery. Group rats tended to have slightly lower urea III nitrogen and total protein. Total protein was still depressed in Group rats 14 days later. Clinical results are detailed in the Clinical Report. Pathology: No treatoent-related gross or Microscopic pathologic lesions were observed in any animals sacrificed a*- '*--ign intervals. Of the 2 aniacis which died during the test period, tib-ues of only 1 were saved for pathologic examination. Death of this animal was attributed to severe pulmonary edeaa and was concluded to have been r rear Bent-re la red. Pathologic results are detailed in the Pathology Report. Organ and Body Weight Analysis: Mean body weights of Group IT rats were siailar to weights of controls during the exposure period (Fig. 1). Mean body weights of Group TTT rats were significantly lower than controls from exposure 2 through 5. Through the remainder of the exposure and recovery period, rate of weight gain was not significantly different between all groups. A comparison of absolute organ weights and relative organ-to-body weight III ratios between Group TT and controls showed no significant differences following the 10th exposure. Mean liver-to-body weight ratios of Group rats were significantly higher in rats sacrificed after 10 exposures* Following the 14-day observation period heart weight and heart-to-body weight ratio of Group TT rars were significantly higher than controls. Kidney-to-body weight rarios of Group TT rats was also higher. Kidney weight of Group TTT rats was significantly higher than controls. Although "'significantly" different, lack of dose-related trends in these effects makes them difficult to evaluate. ^Panysan^ooesno, GontefnTSCAcei 4 - c*i^'---- Su--ary: Groups of 10 male Crl:CD rats were exposed 6 hours/day, 5 davs/week for 2 weeks to concentracloM of either 1.5 or 9.6 ag/1 ofl B^B1" alr* A conrr0^ group was simultaneously exposed to air only. During exposures, reduced response to Bound was obaerved In all teat rat-s. Rapid breathing waa obaerved In Group HI rata following high concentration excursions' Following expoaure, clinical signa of Group IT rata were indlsrlngulshable from controls. Group TTI rats exhibited severe following 2 exposures - normal weight gain thereafter. Two Group died - 1 found dead on day 2 and the other died during the second weight IOBC TTT rata exposure. Clinical chemistry measureaents of Group II rats were Industingulshable froa controls. After 10 exposures. Group TTI rats had slightly lower values for urea nitrogen and total protein. The total protein waa also lower In Group TTT rats following the 14-day recovery period. No treatment-related gross or microscopic pathologic teutons were observed In any animals sacrificed at design Intervals. Of the 2 aolaals which died during the test period, tissues of only 1 were aaved for pathologic examination. Death of this aniaal was attributed to severe pulmonary edema anil was concluded to have been treacaent related. Mean body weights of Group TT rats were similar to the controls throughout the test period. Group TTT rats mean body weight was significantly lower than f-he controls on days 2 through 5 of the exposure period. The rate of weight gain of Group TTT fats was similar to the controls through the remainder of the rest period. No significant differences were observed in organ/body weights of Group TT rats sacrificed after 10 exposures. Mean liver weight of Group ITT rats was significantly higher than controls after 10 exposures. Company Sanitized. Does not contain TSCA CBI (1) Walter B. Konlecki and J. R. Clinical Laboratory Reporr," ^J^^^H Barnes, "Subacute Inhalation ^----------H-13,383,Septeaber 9.I9BU^^ ,^H|B> (2) Ravnond M. Evererr and UllllaB C. KrauBS, Haakell Pathology Report No. 46-80 H-13,383, October 24, 1980. Report by: ^ / ^ yV f StephHeenn D. Nah Terhnlclan Approved by: a^^.. ^~ ^ 'r Gerald Li.. Knnnneeddy Chief, Acute TnvesriIgarilons Section SDN:vln Study Director: R. L. Ferenz Date Tssued: February 24, 1981 Reoorr No. 88-81 .Company Sanitized. Does not contain TSCA CBI 6 - l-U > 0^, Z3 C-J Q CK: C-3 h- (-3 Il-U ^ - :-3 C_) <^ no -^) LO FIGURE I !" a u* d - ^^ id Q 5 s 07 Company Sanitized. Does not contain TSCA cfe APPENDIX I Company Sanitized. Does not contain TSCA CBI 08 (2-UK SUBACUTE BODY WEIGHT SUMMARY H* 13383 GROUP CON'rRCLS LOW LEVEL(;1 MO/L) HIGt^ LEVEL)no MG/L) F RATIO;!) L.3D(2) I l'JNNETT(3> h1.1S<4) INTERVALS IN TEST DAYS 1 2 3 236.8000 238.8000 2383770 236.8000 239.300C 217.250v 2tH.7000 2' >4.2000 22.8.5000* 0.980 3.2029 3.6547 11.1630 28.676* 6.3334 7.2268 43.6480 15.391* 3,0.1449 1,1.5758 11,1.9880 4 249.,8000 2SI.,9000 231.,3750* 12(.270* 8. 7071 9. 9352 82. 4950 5 2S4.>40GO 257..3000 240..3750* 1J1,955* 7..5188 8. S794 61. 5150 GROUP i INTEFtVALS IN TEST M iYS 8 9 10 11 12 9L3W CONTROLS LEVEL <I MG/L) HIGH LEVEL< 10 MG/L) F RATTIIOO((1I) LSD<2) DUNNETT(3> UHS(4) 272.1000 274*4000 266.8750 2*1082 7.3814 .4226 59.2870 28U^^ 279*39'00 28 4.7000 ' 291.4000 27AWSO "" 27j|.2500 . ; ' -^-siZis^^:,-^:^: r ^-00..9924 :' ^.wo':^. . I* 1072 7^7375' ',,. 5p.03SO^- ^Tt.-52250008 286. 5000 287. 8000 282. 3000 11.155 7.2455 8.2675 57.1240 289. 7000 268. 20CO 265. 2500 11.181 5*9008 6*7331 37.8880 GROUP INTERVALS IN TEST DAYS IS 14 17 CONTROLS LOW LEVELd MG/L) HIGH LEVEL(10 MG/L) F RATIO(l) LSD<2) DUNNETTO) UMS(4) 305.8000 305.0^00 302.0000 0.152 15.4624 17.77 6 113.8909 310.8000 308.0000 " )9.7500 0.099 14.5384 16*7116 100.6864 314.2000 312.6000 314.2500 0.042 14.4937 16.6602 100.0682 18 317.8000 317.2000 318.7500 0.026 14.5830 16.7628 101.3045 19 322,6000 319*6000 322.0000 0.118 14.8427 17.0613 104.9455 .Company Sanitized. Does not contain TSCA CBI 09 G^OUF ,INTE RVAA5 IN TES T DAY 5 i 23 24 25 26 CONT ROLS LOW LEVEL( 1 MG/L) HIGH LEVEL ( 10 M5/L) 345.0000 340>6000 343.2500 347. 6000 343. 8000 349. 5000 351 .2000 348 .4000 353 .5000 3SO. 2000 345* 8000 357. 5000 F RATIO<1> LSD<2> DUNNETT(3) UMS(4) 0.105 22*0943 25*3969 232.5409 0 .167 22. 0643 25. 3624 231. 9091 0.129 21 .8383 25 .1026 227 .1818 0 .623 22. 7104 26. 1051 245. 6909 (1) RATIO OF AMONG- TO WITHIN-SROUP VARIATION--ONE-FACTOR ANALYSIS OF VARIANCE. ;2> LEAST SIGNIFICANT DIFFERENCE--GIVEN A SIGNIFICANT (ALPHA0.05) F RATIO ANY TUO MEANS DIFFERING BY MORE;THAN THE LSD ARE SIGNIFICATLY DIFFERENT WITH A VARIAhLE-UISE FALSE POSI+IVE <ALPHA? ERROR RATE OF 0.05. ' i . . <3) DUNNETT rEST--ANY TREATMENT MEAN DIFFERING FROM THE CONTROL MEAN BY MORE THAN THE DUNNETT STATISTIC IS SIGNIFICANTLY DIFFERENT FROM THE CONTROL MEAN WITH A VARXftFLE-MISE FALSE POSITIVE (ALPHA) ERROR RATE OF 0.05. (4) WITHIN-GROUP MEAN SQUARE, + SIGNIFICANTLY DIFFERENT (P<0J05) FROM CONTROL,'GROUP BY LSD. * SiJ ^^ SIBNIFICANTLY DIFFERENT <P<0.io5) FitOM CONTROL-IBROUPBY DUNNETT TEST AND LSD. ^: SIGNIFICANTATTHEC.05PROBAiBILrTYI.EVeL, , ; . ''-,. '':. 'i: Company Sanitized. Does not contain TSCA CBI 10 yff^w^^~'-':-.^^--^'-'^---T-'---.7^y .".- --- ' ~^^"?Bt3.:"''"5Il'!^^a%^'s^ APPENDIX II Company Sanitized. Does not contain TSCA CB1 11 2-lx'K SUBACUTE ORGAN/BODY WEIGHT ,1EAN ABSOLUTE DATA H* 13383 RATS SACRIFICED AFTER 10 EXPOSURES GROUP FINAL UGT. HEART LUNGS LIVER SPLEEN CONTROLS LOW LEVEL(1 .0 MB/L) HIGH LEVEL( 10, 0 MG/L) 389,0000 386*4000 284,5000; F RATIO'1> LSD<2) DUNMETT<3) UHS(4> 1.197 7*6042 8*7409 27*5455 1.0260 .9630 1.0700 2.434 .1009 .1160 .0049 1.6B80 1.5820 1.8300 1.225 .3423 .3935 .0558 11.1920 10*6140 12*1325 3.676 1.2297 1.4135 .7203 .6700 .5620 6525 2*994 .1075 .1236 .0055 GROUP CONTROLS LOW LEVEL(1.0 MG/l ^ HIGH LEVELdO.O 1G/L) F RATIO<1) LSD(2) DUNNETT(3) UHS(4> KIDNEY i 2 * 3360 2.3380 ; 2*5775 ! 2*716 .2527 .2905 0504 TEST IS 2*9040 3.0260 3.0250 ^ ^ .3845 ,4430^ y070s4 THYMUS .7740 .7580 .7425 .051 .2144 .2465 .0219 (1) Ratio of amonfl- to within-s(p|oyfry<r(<tifl(c>--one--factor analusis of variance. <2> Lt significant diffrr>ce--^il3tjiir^y sis^if^ant <lfh005> F ratio? ana two irans differina bw tor "f'tiMioilllnll^S hiM>. fanificantlw different with a variable-wise false ^oslti.^l-'^al^^l^/.B'rror^ate of 0.05. (3) Dunrrtt twt--An treatment illfai6^ffepirsl from the control mean by aore than the Dunnett statistic is siati|^ie|intlu different from the control mean uith a variable-Mise falsa positive' <aliha) error rate of 005 <4) Uithin-tfroup Mean Sauare. I + Significantly different (P<0OS) froB control flroup by LSD* # Sianificantia different (P<0.0fe) froift control group b Dunnett test and LSD* ! Company Sanitized. Does not contain TSCA CB1 18 GROUP 2-UK SUBACUTE ORGAN/BODY WEIGHT MEAN RELATIVE D^TA H# 13383 RATS SACRIFICED AFTER 10 EXPOSURES HEART, LUNGS LIVER SPLEEN MONEY CONTROLS LGU LEVEL<1.0 MG/L) HIGH LEVEL(10 .0 M6/L) F RATIO(1) LSIK2) DUNNETT(3) UnS<4> .3541 * 3381: .3765 2.391 .0380 .0437 .0007 .5828 5S28 .6423 i .341 , 190 .1368 .0068 3.8602 3.7049 4.2743+ 4.614* .4133 .4731 .0814 .2311 .1960 .22PS 3.016 .0364 .0418 .0006 8059 .81S7 .9064 4.085* .0822 .0945 .0032 GROUP TESTIS: THYMUS CONTROLS LOU LEVEL(1.0 MG/L) HIGH LEVELdO.O MG/L> F RATIO(l) LSD<2) DUNNETT(3> UriS(4> 1*0029; 1.0560 1.0629 637; .1302' .1497' 0081 ; 2675 .2649 .2607 .619 .0758 *0872 0027 fair (1> Ratio of aaons- to within-a!pou> y^ia!;tion---o^ff-'f^cto^ analysis of variance* <2> any Le two ast lea si ns gni al ficant ffTina diff bw vre--Miv(n i idni BpNt.'(.hi|r 't^fj^^BK:;r9 f icant siani f (al ican h tly 0.05> diffe F pe ra nt t i o f with a vriab'-wi posit iv(lh> error rate of 0*05. <3> Dunnett test--Any tratrni nran difftrina from the control mean by more than the Dunnett statistic is significantly different from the control mean with a variable-wise false foitjive (alpha) error rate of 0*05* (4) Uithin-aroup Mean Sauare. + Significantly different (P^.'QS) fro control aroup bw LSD. # Sianifxceritia differertt <P<0!05> froa control aroup by Dunnett test and LSD. ' Sianifleant at the 0.05 probability level. Company Sanitized. Does not contain TSCA C81 13 2-WK SUBACUTE ORGAN/BODY WEIGHT MEAN ABSOLUTE DATA H# 13383 RAT5 SACRIFICED AFTER 1-4 DAY OBSERVATION PERIOC GROUP FINAL MGT. HEART LUNGS LIVER SPLEEN CONTROLS LOW LVEL( 1 MG/L) HIGH LEVEL ( 10 MG/L> F RATIO(1) L50(2) OUNNETT(3) UMS<4) 350.2000 345.8000 357.5000 .623 22.710-4 26.1051 245.6909 1.1000 1.2300+ 1.1950 2.792 .1298 .1492 .0080 2.0420 2.0460 2.1750 .355 .3811 .4380 .0692 12.S760 12.9560 13.4075 .143 2.2753 2.6155 2.4662 .6760 ,6440 .7200 ,853 ,1257 ,1445 .0075 GROUP KIDNEY' TESTI8 THYMOS CONTROLS LOU LEVELd MG/L) HIGH LEVEL(J10 MG/L) i 2.5700J 2.77801 2.8425* 3.1000 3.1580 3.3175 . 7580 .7000 .7825 F RATIO :!) LSD<2> JPUNNETT(3> UMS(4) 3.729 .2308 .2653' .0254 1*407 .2864; .3292 .^l 428 .2016 .2317 .0194 1) C<2) Ratio of on<- tu within^|wou> vyrtsi't^f's-'-iciw-fscl.or analwcis of variance. Least significant diffepric|e--stivffr 'a ."st^ni^icant <8lpha=0.05) F ratio- ana two means dif^epind baaore^hah the LSiDJ arc^siani fi cant 1 different uxth a variable-wise false ^Qsft^^(9liirf's^ TTor rate of 0.05, (3) Dunnett tt--Any traatineni.! iij(an differi'nai.f.roiii the control 9e-. by more than the Dunnett statitic/ic i-ifbi-^icantiM dif^ from the control aean with a variabie-uise fa2e positjtv (ali-ha) error ^rate of 0.05. (4) Uithin-droup Mean Sauare. , ; ( Siatnificaintia different <P<005) from oentrol tfroup b LSD. Sl^nificantlw different (P<0.05> from control tfroup bw Dunnett test and L50. i Company Sanitized. Does not contain TSCA C8 14 2-WK SUBACUTE ORGAN/BODY UE^HT MEAN RELATIVE DATA H# 13383 RATS SACRIFICED AFTER 14 DAY OBSERVATION PERIOD GROUP HEART LUNGS LIVER SPLEEN KIDNEY CONTROLS LOW LEVELd MG/L> HIGH LEVEL( 10 MG/L> F RAUO<1) LSD<2> DUNNETT<3) UMS<4) .3143 *3559* .3340 4.933 .0304 .0349 .0004 .5829 .3920 .6071 .153 .0949 .1091 .0043 3. 6726 3. 7401 3. 7480 ,060 .5339 .6137 .1358 .1931 ,1862 .2009 .307 ,0315 .0362 .0005 .7331 .S056+ .7957 3*452 .0675 .0776 .0022 GROUP TESTIS THYMUS CONTROLS . LOW LEVEL<1 MG/L) HIGH LEVEL<10 MG/L) F RATIO<1> LSD(2) DUNWETT<3> UMS(4) .8861 .914^ .9290 i .57^ .0890 *l023 .003^ .2170 .2011 .2187 .306 .0554 ,0636 .0015 1) Ratio of sftond- to within-Mroup vsristion-'-pne-factop analysis of variance. <2) Least sidni f leant diL;rfrerjce--fliven a siflnificant (8lpha=0.05) F ratio any two Beans differing bu ii9.rethan the LSD ape sianificantia different with vrisble-wise falsf posi'tiye (alpha) errj&r rate of 0.05. /<T3)' Dunnett test--Any treat--ni ran differinfl fron the control mean by more than th Dunnett statistic ^Silijillftificantl'a difftrent fro the control Biean uith a var.able-wise false pos^tiv (alpha) error rate of 0*05. <4> Mithiri-tfroup Mean Sauare, | ; .^ + Sianificantly different (P<0!05) fro contpei stroup ba LSD. # Sianificantlw different (P<0|.05> from control, stroup by Dunnett test and LSO* i * Significant at the O.OS probability level. Company Sanitized. Does not contain TSCA r*w 15