Document jy4zD8gE5JQDbOvnjvk9kBb1O
AR226-2894
COMPLETE REPORT IN MR FOLDER.
E. 1 du Font de Nemours and Company Haskell laboratory for Toxicology and Industrial Medicine
Elkcon Road, Newark, Dexaware 19711
HASKELL LABORATORY REPORT NO. 88-81
Material Tested
Haskell No 13,383
Stud'1' Initiated/Completed 5/5/80-5/30/80
Material Submitted bv
Eimicals & Pigments Department Jackson Laboratory
SUBACUTE INHALATION TOXICITY STUDY O F i M M --
M P O N JjATS
Introduction: The purpose of this study was to observe the effects on sale
rats of repeated inhalation exposure t o g m H H H M ) vapors. In a previous
study g p | c h e LC30 for
was determined to be 107 mg/1.
Design concentrations for this s t u d y a r ^ ^ ^ m g / 1 (Group I), 1.0 og/1 (Group
II), and 10.0 mg/1 (Group H I ) f ( ^ ^ B B B k i n air.
Procedure:
A. Animals
Male Crl:CD rats were housed in pairs in stainless steel wire mesh cages. Purina Certified Rodent Chow #5002 and water were available ad libitum. All rats were held under these conditions for a 1 week pre-test period to insure suitability.
B. Exposure Protocol
Groups of 10 rats weighing 234.0 to- 245.0 grams were exposed to
atmospheres containing
either 1.0 mg/1 " Group II or 10.0 mg/1 ~
Group III) 6 hours/day, 5 davs/week for 2 weeks. Control raes (Group I)
weighing 234.0 to 242.0 grams jre simultaneously 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.
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C. Generation
Atmospheres of the teat material were generated by syringe driving the
compound through a Spraying Systems nebulizer. The syringe and nebulizer
were heated to a temperature of approximately 70C. The aerosolized material
sprayed onto the heated (50C) surface of a 3-neck round bottom flask.
Houseline air swept the 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 tandem midget impingers 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 20% S.E. 30 on 80/100 mesh Chromosorb W-HP. The G.C. oven temperature was programmed with an initial 2 minute hold at 50C, increased at 32 /minute to 2 0 0 and held there for 2 minutes
G.C. analysis was based on tl._ f^hbomponent. This peak was identified
on the basis of the G.C* peak height area and the reported sample
composition. Sample concentrations were determined by comparison with
standards prepared by dilution of the test compound. Chamber concentrations
were expressed as the equivalent
concentration (mg/1).
Oxygen and chamber temperature were monitored throughout each exposure. E. Clinical Measurements
Clinical laboratory measurements were made on urine samples collected overnight following the 9th exposure and the 13th day of recovery. Blood samples were taken from the rats* tails after the 10th exposure and the 14uh day of recovery. Detailed clinical chemistry procedures and indices are outlined in the Clinical Chemistry report (1).
F Pathology
After the 10th exposure, 5 rats from each group were selected at random and sacrificed for gross ind histopathological examination. Remaining rats were sacrificed on the 14th day of recovery for an identical examination. Pathological procedures and indices are included in the Pathology reoort (2).
G. Organ and Bodv Weight Analysis
Organ weights and organ-to-body weight ratios were determined for the heart, liver, lungs, kidneys, spleen, testes, and thymus (Appendices I and
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Result: high bo
During exposures, low boiling components evaporated readilv while ng solids plugged the generation apparatus. This problem accounts
I t l , ?ard devlacon and range for these exposure concentrations
Detailed data is listed in following Tables.
cions.
GROUP II
Design Level - 1.0 mq/1
Exposure No.
1 2 3 4 5 6 7 8 9 10
Average (mg/1)
2.02 1.77 0.99 1.44 1.91 1.11 3.58 0.97 o.-: ' 0.78
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 0.23-2.74 0.14-8.09 0.14-6.23 0.16-5.31 0.08-8.26 0.27-2.25 0.14-2.17 0.0-1.68
GROUP III Design Level - 10.0 pg/j
Exposure No.
1 2 3 4 5 6 7 8 9 10
Average (me/l>
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
Ranee
3.7-48.50 1.8-16.0 2.3-17.3 2.9-9.8 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 J 5 + 1.9 (Group II) and 9.6 + 6.4 (Group III)
Chamber temperature wais jC28C during all exposures. ^20^ during all exposures.
Chamber oxygen
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3-
zszsxf*concentration excursions.
zzs.C^ .n ' < t W W . raduccd
s ^ j r s"l.- -11ce-r n-ighr
nmnoraaral tsrateexhoifbiwteedighmtodgeariante. to severe 2 i g8zS 1loosbs8jffroirll?lldowwieenddcbr?y*>*repc1oo,v-uer*ryt oGf roup\
c^sr^lll
tte --
Another
~or? 1f4Cd^ayCs hreetcllosvrrevry: . NGoreoufpfeTc tIsI wraetrse tseenednedin toGrhoauvpe TsI rla tes ha ftle rv l ^ " 1in0
M
which died during the test period. tlb^ e s ^ f orlVl lireLve5 L i
pulmonary edema and was concluded to have been i r e a ^ ^ r ! ? J W *
results are detailed in the Pathology Report.
related. Pathologic
Organ andto BwodevigWhetisghotf Acnoanltyrosilss:duMreinagn rbtoadv. weiah. *ts of Gm..n(F nI " 5*
Sir* ^ ' S S S ^ ^ S S - ^ r : r
^ i S 3 -S5r a s were significanrly higher in rats sacrificed after 10 exposures.P
4
Ti v
V
the. difficult t,, e " " , ; .
do..-t.l.t.d trend. 1,, the.. effect. ,, h e .
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uaMrjr: Groups of 10 Dale Crl:CD0 rats were exposed 6 hours/day. 5 ^ ^ g w e e k for 2 weeks to concentrations of either 1.5 or 9.6 mg/1 ofk j g g g ln alr* A control group was simultaneously exposed to air only?
J e*po#ureB* reduced response to sound was observed in all test
rats. Rapid breathing was observed in Group TIT rsts following high
concentration excursions.
"** "
Following exposure, clinical signs of Group II rats were indistinguishabie iron controls. Group TII rats exhibited severe weight loss following 2 exposures - normal weight gain thereafter. Two Group III rats died - 1 found dead on day 2 and the other died during the second exposure.
f r n m Cli"lr*1 `henistry measurements of Group II rats were induetinguishable fron controls. After 10 exposures, Group III rats had slightly lower values for urea nitrogen and total protein. The total protein was also lower in Group ITT rats following the 14-day recovery period.
No treatment-related gross or microscopic pathologic lesions were observed in any animals sacrificed at design intervals. Of the 2 animals which died during the test period, tissues of only 1 were saved for
pathologic examination. Death of this animal was attributed to severe pulmonary edema and was concluded to have been treatment related.
Mean body weights of Group TT rats were similar to the controls throughout the test period. Group TTI rats mean body weight was
significantly lower than the controls on days 2 through 5 of the exposure period. The rate of weight gain of Group TIT rats was similar to the controls through the remainder of the test 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.
-5-
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(1) Walter B. Koniecki and J. R. Barnes^Subacute Inhalation of^
Clinical Laboratory Report,"
H-13,383, Septeaber 9,
(2) Raymond M. Everett and William C. Krauna, Haskell Pathology Report No. 4 6 - 8 0 H-13,383, October 24, 1980.
Report by:
Technician
Approved by:
f- *
--- jf*
Gerald L. Kennedy
Chief, Acute Investigations Section
S D N :vim Study Director: R. L. Ferenz Date Issued: Februarv 24, 1981
Report No. 88-81
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SUBACUTE WGT. GROWTH CURVE
07
eupi-.
APPENDIX I
*
i i
?)
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r>-UK* SUBACUTE BODY WEIGHT SUMMARY H* 13383
GROUP
INTERVALS IN TEST DAYS 12 3 4 5
CONTROLS LOW LEVEL(1 MG/L) HIGH LEVEL(10 MG/L)
F RATIO i1) L3D(2) DUNNETT(3 > WMS(4)
236 >8000 238.8000 238>37*0
0.980 3.2029 3.6547 11.1630
236.8000 239.3000 217.25004
28.678* 6.3334 7.2268 43.6480
241.7000 244.2000 218.5000*
15.391* 10.1443 11.5753 111.9880
249.8000 251.9000 231.3750*
13.270* 8.7071 9.9352 82.4950
254.4000 257.3000 240.3750*
11,955* 7.5188 8.5794 61.5150
GROUP
CONTROLS LOW LEVEL(1 MG/L) HIGH LEVELdO MG/L)
F RATIO(l) LSD(2) DUNNETT(3) WMS(4)
GROUP
. "i
INTERVALS IN TEST DAYS
8< 1
9
10
272.1000 279'.3000 274.4000 ! 281.1000 266.8750 i 274.3750
24,7000
282-.4000 277.2500
2.102 I ' 7.3814 ! 6.7810
S.4226 59.2870
7.7375 50.0350 v
. j'
-0.924 #.1072 :?2508 71.5200
i 1 '
INTERVALS IN TEST DAYS
15 ! i
16
17
CONTROLS LOW LEVEL(1 MG/L) HIGH LEVELdO MG/L)
F RATIOd) LSD<2) DUNNETT(3) WMS(4)
305.8000 305.0000 302.0000
0.152 15.4624 17.7/ 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
-iJ*.
286.5000 287.8000 282.5000
1.155 7.2455 8.2675 57.1240
18
317.8000 317.2000 318.7500
0.026 14.5830 16.7628 101.3045
12
289.7000 263.20C0 265.2500
1.181 5.9008 6*7331 37.8880
19
322.6000 319.6000 322.0000
0.118 14.8427 17*0613 104.9455
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09
GROUP
23
24 .
25
CONTROLS LOW LEVEL(1 MG/L) HIGH LEVEL(10 MG/L)
F RATIO(l) LSD<2> DUNNETT<3> WMS(4)
345.0000 340.6000 343.2500
0.105 22.0743 25.3969 232.5409
347.6000 343.8000 349.5000
0.167 22.0643 25.3624 231.9091
351.2000 348.4000 353.5000
0.129 21.8383 25.1026 227.1818
350.2000 345.8000 357.5000
0.623 22.7104 26.1051 245.6909
! " R A T 1 F m m > - T U IT H IN - R UP V AR IATIO N " --ONE--FACTOR A N A L Y S IS OF V AR IAN C E.
ANY TWO HEANS ^ D IF F E R ! NG^BY^MRE^THAN^THE L S n I2 > r F 5',N T ` A L P H A -O .O S J F R A T IO . ITH A VARIABLE-UISE FALSE P O S ltlV E
THEN^ N Er ? r S- ^ i i Si Pr i f ^ GNM^ i c A ; n r i ? F ^ ^ " T TF ^ ^ " NTR0L * * IT H A V A R IA F LE - N IS E FALSE P O S IT IV E X E S t f ERROR RATe Sf V . ' TM TM "
<4) W ITH IN-G RO U P MEAN SQUARE.
HORE
t SIONIFICANTLY DIFFERENT <P<0.:0S> FROH CONTROL SROUP SY LSD.
# C E R E N T <P<0.i>3 , FROH C O N T R O L * O U P SY DUNNETT T E S T AND L S D .
S I G N I F I C A N T AT THE 0.05 PROBa Ib I L I T T L E V E L .
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APPENDIX II
#
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11
,- J
2-wk s u b a c u t e o r g a n /bod y w e i g h t ,1EAN a b s o l u t e d a t a I H* 13383 RATS SACRIFICED AFTER 10 EXPOSURES
GROUP
FINAL WGT.
HEART
LUNGS
LIVER
SPLEEN
CONTROLS
289.8000
LOW LEVEL(1.0 MG/L) 286.4000
HIGH LEVEL(10.0 MG/L) 284.5000;
1.0260 .9680
1.0700
F RATIO'J> LSD(2) DUNNETT(3) WMS(4)
1.197 7.6042 8.7409 ! 27.5455
2.434 .1009 .1160 .0049
1.6880 1.5820
1.8300
1.225 .3423 .3935 .0558
11.1920 10.6140
12.1525
3.676 1.2297 1.4135
.7203
.6700 .5620
.6525
2.994 .1075 .1236 *0055
GROUP
KIDNEY
TESTIS
THYMUS
CONTROLS LOW LEVEL(1.0 MG/l ' HIGH LEVEL(10.0 MG/L)
F RATIO(l) LSD(2) DUNNETK3) WHS(4)
2.3360 2.3380
2*5775
2.716 .2527 .2905 .0504
-
2*9040 3.0260
3*0250 ..;
337 .3845 .4420 .0704
.7740 .7580
.7425
.051 .2144 .2465 .0219
(1) Ratio of among- to within-gr|pup. variation-- one-factor analysis of variance.
(2) Least sianifleant differenceh^giygjn f significant (alPha*0.05) F ratio
ana two naans differing by more thwrit$iiESD ire-iisnificantlw different
with a variable-wise false nositiM J:a^|i^ errpf ^ate of 0.05.
<3) Dunnett twst-- Ana treatment meeh/differing from the control mean by more
than the Dunnett statistic is sighi^ieentla different from the control mean with a variable-wise false positive lelrha) error rate of 0.05.
<4> Uithin-group Mean Souare. |
+ Significantly different (F<0.GS) from control group by LSD.
# Significantly different (PCO.OjiS) from control group by Dunnett test
and LSD.
!
I
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2-WK SUBACUTE ORGAN/BODY WEIGHT MEAN R E L A T I V E DATA H* 133 8 3 RATS S A C R IF IC E D AFT E R 10 EXPO SU RES
GROUP
HEART,
LUNGS
LIVER
SPLEEN
KIDNEY
CONTROLS LOW LEVEL<1.0 MG/L) HIGH LEVEL<10,0 MG/L) *
F RATIO(1> LSD<2) DUNNETT<3> WMS <4>
.3541 .3381'
.3765
2.391 .0380, .0437 .0007
.5828 .5528
.423
-341 . 190 .1368 .0068
3.8602 3.7049
4.2743+
4,614* .4133 .4751 .0814
.2311 .1960
.2295
3.016 .0364 .0418 .0006
,8059 .8157
.9064
4.085* .0822 .0945 .0032
GROUP
CONTROLS LOW LEVEL<1.0 MG/L) HIGH LEVEL<10.0 MG/L)
F RATI0<1> LSD(2) DUNNETT<3> WMS <4)
TESTIS;
1.0029; 1.0560
1.0629
633 < 1302! .1497 ! .0081 :
THYMUS
.2675 .2649
.2607
.019 .0758 .0872 .0027
(1) Ratio of among- to withih+sroue vari#tion-- one-factor anala3is of variance.
<2> Least significant diffei^MW*T^ivj|i*a\-Significant Cslrha-O.OS) F ratio
ana two means differing by mpr* PtWj^i t h a L S D a r e significantly different
with a variable-wise false POiiive(alrh*> error rate of 0.05.
<3> Dunnett test-- Any treatment! mean differing from the control mean by more
than the Dunnett statistic is si[gnifiein$ly different from the control mean
with a variable-wise false rositjive (alpha) error rateof0.05.
(4) Within-group Hean Sauare. !
+ Significantly different <P<0.!05) from control groupbyLSD.
# Significantly different <P<0.!5) from control groupbyDunnett test
and LSD.
;
* Significant at the 0.05 probability level.
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GROUP
CONTROLS LOW LEVEL(1 MG/L) HIGH LEVEL d o MG/L)
F RATI0<1> LSD<2) DUNNETT <3 ) UMS<4)
""
final WOT.
----- --
heart
________________ _____
350 200Q
345.8000 357. 5000
1.1000 1.23004 1.1950
62? 22,7104 26.1051
2456?09
2.792 1298 ' .1492
0080
C U N uS
2.0420 2.0460 2.1750
355 3811 4380 .0692
GROUP
CONTROLS LOU LEVEL<1 MG/L) HIGH LEVEL<10 MG/L)
F RATIOil) LSD<2) DUNNETT(3 ) WMS(4)
kidney-
testis
2.5700i 2.7780! 2.8425#
j
3.729 2308 2653! .0254 (.
3.1000 3.1580 3.3175
1.407 *2864 .3292 .0391
ththus
.7580 7000 7825
428 2016 .2317 0194
mta
liver
12.8760 12.9560 13.4075
.143 2.2753 2.6155 2.4662
SPLEEN
6760 6440 7200
853 1257 1445 0075
ith - r v . 3 S r ^
;, s s S i
th. ,, i s
r a s S K S ^ ^ * ? i ' `^ .
,, a "...., , t e a * a ~ s ~ s a t .
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14
! ^ 2 CUTE ORGAN/BODY WEIGHT MEAN RELATIVE BATA H# 13383 RATS SACRIFICED AFTER 14 DAY OBSERVATION PERIOD
GROUP
HEART
LUNGS
LIVER
SPLEEN
KIDNEY
CONTROLS LOW LEVEL(1 MG/L) HIGH LEVEL(10 MG/L>
F RATIO(1) LSD(2) DUNNETT(3) WMS(4)
.3143 .3559* .3340
4.933 .0304 .0349 *0004
.5829 .5920 .6071
.153 .0949 .1091 .0043
3.6726 3.7401 3.7480
.060 .5339 .6137 1J58
.1931 .1862 .2009
.507 0315 .0362 .0005
.7331 .3056+ 7957
3.482 .0675 .0776 .0022
GROUP
CONTROLS
.
LOW LEVEL<1 MG/L)
HIGH LEVEL(10 MG/L)
F RATIO(l) ' LSD(2>
DUNNETT(3 > WMS(4)
TESTIS
.8861 .9144 929Q
.57# .0890 *102? 0033
THYMUS
.2170 .2011 .2187
.306 .0554 0.636 0015
-
f aeons- to within-rdrpup variation- ^pne-f,,,acvto.r. a,,nqatl3yasiSs OoTf varriiaaince i-*a*t significant difference-- given a significant (3lpha=0.05> F ratio
any two means differing bv more than the LSD are significantly different
i v*ri8ble"wise f*ls* positive (alpha) e m b r rate of 0.05
(3) Dunnett test-- Any treatment mean differina from the control mean by more
than the Dunnett statistic is pianificanti$ different from the control mean
with a verable-wise false positive (alpha) error rate of 0.05.
(4) Within--group Mean Sousre j
*
+ Sisnificantly different (P<oj*05) from control group by LSD.
* if2ificsmUw different (P<0j.05> from control Sroup by Dunnett test
and LSD*
I
* Significant at the 0.05 probability level.
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