Document g5BMNEjE0GmY3mdz1jkgd7KQ
HAZLEOCON LAE30RATORiES AJVERICA. INC.
Chemical& BloMedIcaSlciencesDivision
3301 KINSMAN BLVD. P.O.BOX 7545 MADISON, WISCONSIN 53707 PHONE (608)241-4471 TLX 703956HAZRAL MD& UD
REPORT OF ANALYSIS
W. C. MCCORMICK MINNESOTA MINING AND TOXICOLOGY SERVICES ST. PAULP MN 55101
MANUFACTURING
SAMPLE: T-3609 PURCHASE ORDER NUMBER: T351371
SAMPLE NUMBER: DATE ENTERED:
REPORT PRINTED:
407039c, 07 /19/E 10/05/E
ENCLOSED:
GUINEA PIG MAXIMIZATION RAW DATA APPENDIX
METHOD, SUMMARY
SIGNED:
STEVEN M. GLAZA STUDY DIRECTOR ACUTE TOXICOLOGY
BY AND FOR HAZLETON LABORATORIES AMERICA, INC.
RAW DATA FOR THIS STUDY ARE KEPT ON FILE AT HAZLETON AMERICA, INC., MADISON, WISCONSIN.
LABORATORIES
HAZLR;OCCN LABORATORIES AN@ERICA IP4r-.
3301 KINSMAN BLVD. 9 P.O. BOX 7545 o MADISON, WISCONSIN
53707
Chemical & BloMedical Sciences Division
PHONE (608)241-4471 9 TLX 703956 HAZRAL MDS@UD
SAMPLE NUMBER: 40703984 SAMPLE: T-3609
PAGE
GUINEA PIG MAXIMIZATION
Test Animal: Young adult male albino guinea pigs of*the Hartley strain were procured, maintained individually in 5tainless-steel caaes in temperatureand humiditv-controlled rooms, provided continuous access to Purina Guinea Pia Chow an@ water and held for an acclimation period of at least 7 days. Th; animals were identified by animal number and corresponding ear tag.
Test System: Fourteen male albino guinea pigs, weighing between 443 g and 549 g, were chosen at random for this study. The animals were divided into.two groups consisting of a test group of 10 animals and a naive control group of four animals. A common vehicle control group of ten animals, weighing between 473 9 and 553 g, was utilized for this study and Study No5. 40703985 and 40705145.
D05e Range Study: The purp05e Of the range-finding study was to find a
moderately irritating d05e level to U5e. for the topical application
and a 5ubirritating concentration to be Used for the challenge application.
Four additional animals were treated with the test Material at concen-
trations of 25@ 5op 75 and 90% w/v in 80% ethanol/distilled water.
The test mixtures were applied to 2.0 x 2.0-cm Whatman No. 3 filter papers
w hich were applied to the shaved animal5l backs and were then overwrapped
with Blenderm and Ela5tOpla5t
tapes. The dressings remained in place for
24 hours. The test 5ites were evaluated for dermal irritation 24 and
48 hours after bandage removal. Based upon the results obtained, the
test material was administered as a 90% w/v suspension for the topical
and challenge applications.
Preparation of Test Material:
Intradermal Injections
Solution No. 1: The Freund'5 Complete Adjuvant Solution was prepared bv adding 3.0 ml of sterile water for injection in increments io 3.0 ml of Freund's Complete Adjuvant.
Solution No. 2: A 5% w/v mixture was prepared by weighing 0.50 g of T-3609 into a 30-ml beaker and adding 80% ethanol/ distilled water to a total volume of 10.0 ml.
Solution No. 3: A 5% w/v mixture of test material in Freund'5 Adjuvant was prepared by adding 2.0 ml of a 10,% W/v suspension of T-3609 in 80% ethanol/di5tilled water in increments to 2.0 ml of Freund's Complete Adjuvant.
For the topical induction and challenge application, the test material was administered as a 90@'a W/V suspension in 80% ethanol/distilled water.
All test mixtures were thorouahly stirred durina preparation and dosing.
HAZLETON
LABCFTATORIES ATIERICA, INC.
3301 KINSMAN BLVD. * P.O.BOX 7545 * MADISON, WISCONSIN 53707
Chemical& BloMedicaSlciencesDivision
PHONE (608)241-4471 TLX 703966HAZRAL
c,AMPLE NUMBER: 40703984 SAMPLE: T-3609
PAGE
3
GUINEA PIG MAXIMIZATION
(CONTINUED)
Preparation of Vehicle Control Mixtures:
Intradermal Injections
Solution No. 1: The Freund'5 Complete Adjuvant Solution was prepared by adding 3.0 ml of sterile water-for injection in increments to 3.0 ml of Freund's Complete Adjuvant.
Solution No. 2: A 5% w/v mixture of 80% ethanol/distilled water in sterile water was prepared by adding 0.5 g of 80% ethanol/distilled water into a 30 ml beaker and adding sterile water for injection to a total volume of 10 ml.
Solution No. 3: A 5% w/v mixture of 9096 ethanol/di5tilled water in Fraund'5 Adjuvant was pr.epared by addina 1.5 ml of a 10% w/v mixture of 60% ethanol/distilled water in sterile water for injection in increments to 1.3 ml of Freund's Adjuvant.
For the topical induction and challenge application,, the vehicle control animals were treated with 80% ethanol in distilled water.
All vehicle control mixtures were thoroughly stirred during preparation and dosing.
Method: A 4.0 x 6.0-cm area was clipped along the midline over the shoulder region. Two rOW5 Of three-deep intradermal injections (total of six injection sites) were made within the boundaries of a 2.0 x 4.0-cm area, one row on each side of the midline as follows per site: 0.05 ml of the prepared Freund's Adjuvant Solution, 0.05 ml of the 5% w/v test mixture, and 0..05 ml of the E;% wlv tes.t material in Freund's Complete Adjuvant mixture. The respective vehicle control mixtures were administered to the vehicle control animals in a similar manner.
Because the test and vehicle control material5 were non-irritating, the areas of the intradermal injections were pretreated with a 10*,iw/to 5odium laurvl sulfate (SLS) suspension in petrolatum 24 hours prior to topical application. Six days after the intradermal injections the test area was closely shaved and the 10% SLS was massaged into the skin with
a glass rod. Twenty-four hours after pretreatment a 2.0 x 4.0-cm patch of Whatman No. 3 filter Paper was saturated with a 90,% w/v suspension of T-3609 in 80,% ethanol/distilled water, placed over the injection
sites, then covered with an overlappina of eienderm tape and secured by
an overwrap of Elastoplast tape. The @ressing remained in place for 48 hours. The vehicle control animals were dC5ed in the same manner
using 80% ethanol/distilled water only.
HAZLPmTmON LABORATORIES ANAEi:41CA.INC.
Chemical& B]oMedicaSlciencesDivision
3301KINSMAN BLVD.o P.O.Box 75459 MADISON,WISCONSIN 53707o PHONE (6082)41-447o1TLX 703956HAZRAL MDS-UD
SAMPLE NUM13ER: 40703984
SAMPLE:
T-3609
PAGE
4
GUINEA PIG MAXIMIZATION
(CONTINUED)
Two weeks after topical induction the test and naive control (previously untreated) animals received a challenge do5e. The hair was removed
from a 5.0 x 5.0-cm area on the right flank by shaving as before. The
90% w/v suspension of test material in 80% ethanol/di5tilled was applied to 5aturation on a 2.0 x 2.0-cm piece of filter vehicle control animals were d05ed in the same manner using
water paper. The 80% ethanol,/
distilled
water.. The patch Was 5ealed to the right flank for 24 hours
under strips of Blenderm tape. Complete occlusion was made by an over@orap
with Ela5tODla5t
tape wound around the trunk.
Observations:
Twenty-four hours following patch removal the test 5ite5
were
examined for ervthema and edema.
The 5it85
were examined again at 48 hours
after patch rem@val to detect any weak, 51OWly
developing reactions.
The
test 5ites were shaved 3 hours prior to the 24-hour reading.
The reactions
were scared according to the following .4-point 5Cale;
0 - no reaction
1 = scattered mild,redness
2 =.moderate and diffuse redness
3 - inten5e'redne55'and
swelling
The important 5tati5tiC
in maximization
sensitization, not the intensity.
testing is the frequency of
Pathology: At discarded.
study termination all animals Maximization Ratings
were euthanatized
Sensitization,Rate
0% 1% - 9% 10,% - 39% 40% - 69% 701% - 1001%
Classification
Not a Skin Sensitizer Weak Sensitizer Mild Sensitizer Moderate Sensitizer Strong Sensitizer
and
HAZLEOCON LABC3FtATC)RIES AA/ERICA. INC
3301 KINSMAN BLVD. 9 P.O.BOX 7545 9 MADISON, WISCONSIN 53707
Chemical& BloMedIcaSlciencesDivision
PHONE (608)241-4471 9 TLX 703956 HAZRAL MDS-UD
SAMPLE NumeER: 4070 3984 PAGE SAMPLE: T-3609
GUINEA PIG MAXIMIZATION
(CONTINUED)
Test Animal: Albino guinea pigs
Source: Hazleton Dutchland, Inc., Denver PA
Date Animals Received: 07110 and 07/24/84
Date Test Started: 08/08/84
Date Test
Completed:
Test Material:
T-3609
09/07/84
Dermal Reactions
to Challenge
Application
An i ma 1 Number
24-Hour Score
EO-2117 EO-2118 EO-2119 EO-2133 EO-2121 EO-2122 EO-2123 EO-2124 EO-2125 EO-2126
Average Score
1 1 0 0 0 0 0 1 0 1
0.4
48-Hour Score
Test Gr-oup
1 1 0 0 0 0 0 0 0 1
0.3
EO-2127 EO-2128
EO-2131 EO-2111
Averace Score
Naive Control Group
0
0
0
0
a
0
0
0
0.0
0.0
HAZLR;OCCN LABORATORIES ANAERICA, INC.
Chemical& B[oMedicaSlciencesDivision
3301KINSMAN BLVD.a P.O.BOX 7545a MADISON,WISCONSIN 53707 PHONE (608)241-4471 o TLX 703956 HAZRAL MDS-UD
SAMPLE NUMBER: 40703984 SAMPLE: T-3609
PAGE
6
GUINEA
PIG MAXIMIZATION
(CONTINUED)
Vehicle Control (80% Ethanol in Distilled Water)
Animal Number
24-Hour Score
48-Hour Score
Test Group
EO-2030
0
@o
EO-2027
0
0
EO-2051
0
0
EO-2052
0
0
EO-2053
a
0
EO-2135
0
0
EO-2140
1
0
EO-2054
0
0
EO-2055
0
0
EO-2056
0
0
Average Score
0.1
0.0
General Behavior study appeared one test group EO-2140, which
and Appearance: All of the guinea pigs used in this normal throughout the study period with the exception of animal, EO-2133, and one vehicle control group animal, had diarrhea on day 3 of the study.
Normal body weight gains were recorded for all animals during the study with the exception of one vehicle control and two test group animals which exhibited body weight losses of 3-24 g during the first week of the study. During the last week of the 5tudy, six test, four naive control and five vehicle control animals exhibited body weight losses
of 2-31 9.
Skin Rea ction5 to T-3609: Slight dermal reactions were observed in four test group animals 24 hours following the challenge application. None
of the naive control animals reacted to the challenge application of
the test material. A slight dermal reaction was observed in one vehicle
control animal following the challenge application with 80% ethanol/
distilled
water.
Conclusion:
Based on the results obtained, the test material, T-3609,
is considered to be a moderate skin sensitizer in guinea pigs.
HAZLE'CCN LABC)PATC)RIES At,/ERICA. INC.
3301 KINSMAN BLVD. * P.O.BOX 7545 MADISON, WISCONSIN
53707
Chemical& BloMedicaSlcienceDsivision
PHONE (608)241-4471 - TLX 703956 HAZRAL MDS-UD
SAMPLE NUMBER: 40703984 SAMPLE: T-3609
PAGE
GUINEA PIG MAXIMIZATION
(CONTINUED)
References: 1. Magnusson, B., MD, and A. Kligman, PhD, MD, Allergic Contact Dermatiti5 in the Guinea Pig, Charles C. Thomas Pub., pp. 113-117 (1970).
2. Hitch, R.K., Ilt)ermalSensitization.Study," Pesticide Assessment Guidelines, Subdivision F, Hazard Evaluation: Human and Domestic Animal@s, U.S. Environmental Protection Agency Office of Pesticide and Toxic Substance Series 81-1, pp. 59-62, November 1982.
Guinea Pigs
@--Test Group: Positive Control Group
-r-3(oO9
A))4
Vehicle Control Group
Naive Control Group
Body Weights (g--ams)
-Vehicle
Vehicle
Vehicle
RT No 4107039
u@a4-c
Animal Number 450-
2111 ztig ?.IICI
Z 12-2-
Technician
Date
19951
,5ncn) 5oz
L4'7(e
!S@q 5-7-7
60;1-7-7 ?e5
51'5 s"q3 593 (o(o& (055
'Act'% 503
5'(06(a 66a
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H 61 S-o73 Yq.? 15vs- 6-08
S73 -T'7-5 r
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9-14 1?-21
SP SP
2 Li
Animal Number elo
5 Zi L
Technician
Date
19
5 5i4cl
676
4LIq 4f3
3-5
NA - Not Applicable
F31t.
sw,
61q
Scale Used: VITgot4 13412 MH Scale Used: )(T)-o)i
I- ILI
Scale Used: k-ry-o@i
Scale Used: @Mn
Scale Used: K-rroh
Lg,o iI?
S;t;;t.P
sr
--9/7
Scale Used:
Reviewed by
v
Date
Test Group: Positive Control Group Vehicle Control Group Naive Control Group
Guinea Pigs Daily Observations
RT No. Room No.
T-3(oO9
ik)
Vehicle 807o6k&4L- @Ylel
Vehicle
pf
Voblcle
2111 ?//g
A)
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Ani=Ll Number
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Room No.
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AJA
A)#4
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Vehicle A)/@
Animal gtum=mr E6 -
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Date
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Test Compound Data AnlaiLLs.Rac"vod Data of ratraderml xmj=tiom Data of Topii&L ApplicAtion Date Of Ch&Uda"
-7 9-,el-g
Trm
i,(r5A6-rouo
RT No q070399Y
Ha7-lek4 DU"101)0'
Tacbmiciam Tacbnic:Um
Aui=l to., sez
24 Noun -
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2 Mdwacs and D:LffusaRoi=ss
3 Tuten" tedness and Sw&Llius
RV.VX= 3y:
ATV-: 9-M-SL,
Guinea Pigs Body Weights (grams)
4--'TestGroup: Positive Control Group Vehicle Control Group Naive Control Group
Pr
T-3(gQ2
Vehicle Vehicle
a Vehiel
Animal Number
21 z Z131 2111-
Y?o
(09-/
9
So
6921
(o*7,"@" 7 @q
--).I&
5-97
& /7
eo5q_
4@o
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U)a14cr-
Technician
Date
M14 g-14
r-2/
-sp FtIq
Animal Number
Technician
Date
NA Not Applicable
Scale Used:
Scale Used: Scale Used:
k-ry@-3a /Y,-1)9
Scale Used: L/Tropi
Scale Used:
@7
Scale Used:
Reviewed by
Date
lb,Clnkv.li
Test Group: positive Control Group Vehicle Control Group Naive Control Group
Guinea Piss Daily observations
RT No. el07ONY
Room No.
Vehicle
NA
Vehicle
7--3&0?
Vehicle 8070L@@
@0;5-hZletl
Animal Number
;7131 ;?111
@j
IAVI
A.1 Al
4-
t\l
t%/
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Tachn:iclan
Date
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N - No visibleAbuo=alitias ,IA- Not Applicable
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Guinea Pigs Daily Observations
RT No. e1070399q
Room No.
Test Group: Positive Control Group Vehicle Control Group Naive Control Group
A)RJ& 0 91
Vehicle Vehicle
Vhi.l.Fo,70L@A
Animal Num r
Tech-
Date
niciau 191?
AJ
pj
A)
A)
A)
A)
A)
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1 9-S'
N - No Visible Abnormalities NA - Not Applicable
GUOU pTr,xiina=ON T=
Test C*qMsnd
-r -3&0?
D&" AUIM&IS R=aiV"
Data of Ta=aderma IMIOCCIMS
A)
Data of Topl@ia
A))q
DataofCh&Umw
9-g-ge@
im 30. qO 7039;?
Scu=a
TwJzdxljm
AJ
Technician
A)#
Ani=l Nig-,Sax
24 Bourg
Ught
Left
48 Imrs
tixllc
Rill
Recorded 37
sf
Date
Saarj=gCo":
Q a Swz%mL,No lowt:LCM
2.a scatter" WAU toduses 2 a MdMM&te and D:Lfftseladz=S 3 - Tac.on" lodu"s and Sw*lling
&w.Vl= 3Y:
C.li*st Group: Positive Control.Group Vehicle Control Group Naive Control Group
Guinea Pigs Body gaighes (grams)
q07OStclS-
RT go. C/O70
I'@or
0/
Vehicle N)A
&)A
%@eA U)4d.,f-
Animal NuaWr
Eo-
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Tectiu:Lcian
Date
9w
5-aa 5
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12054 z 5 5L-.LO56
Technician
Date
416
5,asr
jfq(g
1-74
&31
15-qn
r MA - Not Applicable
Scale Used:
ScaleUsed: IkTronI'@'019
Seal* Used: K-rrnvv /6,0/
Scale Used: tcreco)
Scale Used:
ScalUesed:
Reviewed by
Date
IOVK .
anima Pigs Da@Ur Observations
RT tio..Room 1*.
Test Ormp.*
fteltic"oatagararap
VOUCIS
A)A
mi" coat=elra"
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DA=:
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Guinea Pigs Body Weights (grams)
RT No. q,07093?
Test Group:
Positive Control Group
Vehicle Control Group
A
Naive Control Group
Vehicle Vehicle
Vehicle
IVA
CD
200S
aoo?
Animal Number :P-o 10, tq 9
Technician
Date
5"7 -7 53 5 5'p71
5p 2-i-q4
Animal Number
Technician
Date
NA Not Applicable
Scale Used: KTroy, @@c.al geal1e Usseedd:
Scale Use Scalee Used.:Scale Used: Scale Used: Reviewed by
1.5oig
/yo
Dat e-9. IC(.
Test Group: Positive Control Group Vehicle Control Group Naive Control Group
Ran5e-
Guinea Pigs - Daily Observations
RTNO. Room No.
703'?EY
(oo I?
Vehiclego/060;4@6,00, tomr-@--
Vehicle
&3.4
Vehicle
Animal Number 60 doce ;2009 2010
TechUlcian
Dat a /9 g
tj @3
@i
Al -AJ
ello
Dose, RcL%r,
'I-
cum" PIG 9A;29AMC)N TLST
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;;&09 '7-10-gq-
30=ce
Data of Togiisa
Limt:wn
&)A
Date, of cuu4qp
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Laft
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6
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9/to
it/to
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0 - 3mc=Ll, No R"c=Lcm L a S"ttered %,tLURedness 2 - Mderace and Difftee ledmss 3 - tatense Redn"s and SwelLing
Rv.7=v= 3T:
q -L-1
HAZLC-OCONLABORATORIES 3301 KINSMAN BLVD. P.O.BOX 7545
AMERICA, INC. MADISON, Wl 53707
(608)241-4471
TLX 703956 HAZRAL MDS UD
Guinea Pig Maximization Sample: T-3609
Study No. 40703984
MAYi.51910)5
Sponsor
Minnesota Mining & Manufacturing Toxicology Services St. Paul MN 55101
-
Sponsor's Representative
W. C. McCormick
Contractor
Hazleton Laboratories America, Life Sciences Division 3301 Kinsman Boulevard Madison WI 53704
Inc.
Study Director
Steven M. Glaza
Amendment No. 1 to the Final Report Effective May 13, 1985
This amendment modifies the following portion of the report.
Page 6, Conclusion. To correctly identify the degree of skin sensitization, substitute the underlined for "moderate".
Based on the results obtained, the test material, T-3609, is considered to be a mild skin sensitizer in guinea pigs.
'gteven M. Glaza Study Director, Acute 'Loxicology Hazleton Laboratories America, Inc.
(070it/cja)
"Bate
Chemical & BioMedical Sciences Division
