Document jBOekRpDQonKyvR7ey4g7V7BR
HAZLOMON
LABCFUC%ORIEB ANAERICA. lNr-
Chemical& BlaMedicaSlciencesDivision
3301KINSMAN BLVD. p.o.BOX 7545 MADISON,WISCR SIN53707 PHONE (6082)41-4471 TLX703956HAZRAL MDS UD
FINAL REPORT
AM@AYY iI c i
..ILLAS D. ZIMMERMAN NNESOTA MINING & MANUFACTURING IXICOLOGY SERVICES PAUL, MN 55101
COMPANY
IMPLE: T-3727 IRCHASE ORDER NUMBER: T357842p REL. 4505
SAMPLE NUMBER: SAMPLE ENTERED: REPORT PRINTED:
50202473 .02/15/85 05/07/85
ENCLOSED:
ACUTE ORAL TOXICITY - METHOD., SUMMARY, PATHOLOGY@ PRIMARY DERMAL IRRITATION - METHOD- SUMMARY PRIMARY-EYE IRRITATION - METHOD, SUMMARY QAU REPORT RAW DATA APPENDIX
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.P MNDISON, WISCONSIN.
LABORATORIES
HAZLOMON
LABOPCWMIES
AAAERICA. INM
3301 KINSMAN BLVD. o P.O.BOX 7545 MADISON, WISCONSIN
53707
Chemical & BloMedicalSciences Division
PHONE (608)241-4471 TLX 703956 HAZRAL MDS Ui)
IMPLE NUMBER: 50202473
IMPLE: T-3727
PAGE
2
'CD ORAL SCREEN
Objective: To determine the acute oral toxicity produced when a test material is administered by oral gavage to rats according to the Organisation of Econom*ic Cooperation and Development's Gui.dolines for Te:5ting-Chamicals,, Section 401,, Acute Oral Toxicity., adopted May 12, 1981.
Test Material: T-3727 Physical Description: Off-white waxy solid Stability of Test Material: Sponsor has purity and stability determinations on file.
Test Animal: Young adult male and female albino rats (approximately 7 weeks of age) of the Sprague-Dawley strain were procurady maintained in group cages in temperature- and humidity-controlled quarters., provided continuous acce sa to commercial laboratory food and water, and held for an acclimation 'period of at least,7-days.'
Acclimated animals were chosen at random for the study. Test animals were housed by sex in groups of five and identified by animal number and corresponding ear tag. Food and water' were available Ad libitum throughout the study, except for an overnight period just before test material administration when food, but not water.,was withhold.
Reason for Species Selection: The rat is the animal classically used due to its small size, ready availability, and large amount of background data.
Method: Five male and five,femals rats weighing between 200 and 298 g were used for each dosage level. The study consisted of four dosage levels (0.20, 0.50y 2.00 and 5.00 g/kg).
Preparation and Administration of Test Material: For each dose level.,the test material was mixed with corn oil and heated in a water bath to form a uniform suspension at a specified concentration. Each suspension was allowed to cool prior to dosing. An individual dose was calculated for each animal based upon its fasted body weight and was administered by gavage. The dose volume of each test mixture was 10.0 ml/kg of body weight.
HAZLOCON
LABORMURIES
ANIERICA. INC.
3301 KINSMAN BLVD. 9 P.O.BOX 7545 P IDISON, WISCONSIN
53707
Chemical & BloMedicalSciences Division PHONE (608)241-4471 TLX 703956 HAZRAL MDS UD
.,.iMPLE NUMBER: 50202473 IMPLE.* T-3727
PAGE
3
--CD ORAL SCREEN
(CONTINUED)
Observations: The animals were observed for clinical signs and mortality at 1. 2.5 and 4 hours following test material administration. The animals were observed daily thereafter for 14 days for clinical signs
and twice daily for mortality.
All animals were weighed just before test material administrationy at 7 days and at study termination. At the and of the study an acute oral LD50 was calculated for each sex.
Pathology: At study termination surviving animals were outhanatized. Animals which died during the study or were euthanatized received a gross necropsy examination and all abnormalities were recorded.
lwqHwA@Z-L4E1ZON
LA13CXROM'C@RIES APVIERICA. lt@,C.
3301KINSMAN BLVD. 9 P.O.BOX 7545 MADISON, WISCONSIN
53707
Chemical& SloMedicalSciericeDsivi'sion PHONE (608)241-4471 TLX 703956 HAZRAL MDS UD
NUMBER: 50202473 IMPLE: T-3727
PAGE
4
:CD ORAL SCREEN
(CONTINUED)
SUMMARY
Test Animal: Albino Rats Sprague-Dawley strain Source: Harlan Sprague-Dawleyp Madison Wi Date Animals Received: 01/22, 02/19 and 03/19/85
Temperature and Humidity of Animal Room: 21 to 25 Degrees C.; 42 to 54% Relative Humidity
Vehicle: Corn oil Method of Administration:
Oral Gavage
Date Test Started: 03/01/85
Date Test Completed: 04/09/85
Estimated Oral LD50*:
Male - 0.28 g/kg ofibody weight 95% Confidence Limits@of 0.15 to 0.51 g/kg Female - 0.43-g/kg of body weight 95% Confidence Limits of 0.19 to 0.97 g/kg
Mortality Summary (Number of Deaths)
Dosage .Level
(g/kg)
Hours 0 -4 MF
11 2 MF M F
0.20 0.50 2.00 5.00
0 0 00 00 0 0 00 10
0 0 23 3 2 0 0 32 2 3
Dosage Level (g/kg)
Male
0.20 0.50 2.00 9;.00
Female
0.20 0.50 2.-00 5.00
Days
3
4
MF MF
00 10 ---
0. 0 11 ---
Average Initial
262 271 254 249
225 226 242 229
5
6
M.F M F
7-14 MF
Total M F Both
00 a0 ---
0a 00
---
10 22 --
--
1/5 0/5 1/10 5/5 3/5 6/10 5/5 5/5 10/10 5/5 5/5 10/10
Body Weights (g) Day 7 Terminal
277
236 ---
---
346 -----
---
220
189 -----
240 220
---
*Thakur., A. K., and W. L. Fazio, 1981. A computer program for estimating LD50 and its confidence limits using a modified BehrensReed-Muench cumulant method. Drug and Chemical Toxicology 4 (3)
297-305.
dab %7
HAZLEZON
LAE3C3FtA7C3MEB AP.IERICA.It-Jr-
-
Chemical& BloMedicaSlciencesDivision
3301KINSMANBLVD.o r-.B0O.X7645 MADISONW,ISCONSI5N3707 PHONE(6082)41-447T1LX70395H6AZRALMOS UD
IMPLENUMBER:50202473 IMPLE,. T-3727
PAGE
5
7CD ORAL SCREEN
(CONTINUED) Clinical Signs
Hours 1.0 2.5 4.0
Days 1 2 3 4 5 6 7 8 9 10 11 12 13 14
Dosage Level 0.20 g/kg.
Males
Appeared normal -5
5
5
00 00000 aa 0
Diarrhea
0
0
0
2 0 .0 1 1 1 0 : 0 0' 0
Dark/red/brown-stained anal/
genital area 0
0
0 @4 4 4 4 4 4 3 3 2 2
Red-stained face 0
0
0
1 433 33 0 0 1 1
Ocular discharge 0
0
0
0122220 00 0
Hypoactivity
0
0
0
3 3 3 4 4 4 0- 0 0 0
Ataxia
0
0
0
0 0 0 2 2- 2 0 0 0 0
High Carriage
0
0
0. 0 0 0 0 a 0 3 1 0 0
Hypersensitivity
to touch
0
0
0
0 000000 11 0
Hyperactivity
0
0
0
000000111 0
Clanic convulsions
0
0
0
000000000 1
Alopecia in abdominal
region
a
0
0
0 0 0 0 0 .0 4 4 4 4
Red-stained abdomen
Piloerection
0 .0
0
0
0
0
111
1 10a0 0
001111 000 0
Yellow-stained
. genital area 0
0
0
1112 22 111 1
Swollen genitals 0
a
0
00011 1000 0
Prostration Death
0
0
0
0000 0000 0
0
0
0
00000 0000 0
00 00
21 10 00 1- 0 10 10
00 a0
00
44
00 00
11 a0 00 1* 0
00 00
11 00 0a 00 0a 00
a0 0.. 0
00
44
00 00
11 00 00 00
.*Animal died in p.m.
HAZLEZON
L&SOF&WOCUBB
AN4ER$r-A, INM
3301KINSMAN 13LVD.9 P.O.BOX 7545 MADISON, WISCONSIN 53707
Chemical & BloMedical Sciences Division PHONE (608)241--3471TLX 703956HAZRAL MDS UD
IMPLE NUMBER: 50202473 IMPLE: T-3727
PAGE
6
:CD ORAL SCREEN
(CONTINUED)
Clinical Signs (continued)
Hours 1.0 2.5 4.0
Days 1 2 3 4 5 6 7 8 9 10 11 12 13 14
Do5aas Level 0.20 g/kg
Appeared normal 5
5
5
Diarrhea
0
0
0
Hypoactivity
0
0
0
Ataxia
0
0
0
Red-stained.face 0
0
0
Dark-stained
anal area
0
0
0
Yellow-stained abdomen/
genital area- 0
0
0
Red-stained
genitals
0
a
0
Alopecia in abdominal
region
0
0
0
Ocular discharge 0
0
0
Piloerection
a
0
0
Females
1 10000344 4 4 4 4 4 200000000 0 0 0 0 0 0 123 44000 0 0 0 0 0 0, 0 0 0 1 1 0 0 0 a 0 0 0 0 4433 33 000 0 0 0 0 0
0 2 2;, 2 2 2 0 0 0 0,@ 0 01 0 0
1 1 1 2 2 2 1 0 .0 0 0 0 0 0 . .:.@
111111000 a 0 0 0 0
000oo
iii i i i i i
0 0 1 1 1 1 0 0 0 0 0 0 0 a'
00011 1000 0 0 0 0 0
1IF9"1
HAZLETON
LABOP,=CR#E! AP.IERICA. INC.
3301 KINSMAN BLVD. n P.O.BOX 7546 MADISON, WISCONSIN 53707
Chemical& BloMedicaSlciencesDivision
PHONE (608)241-4471 TLX 703956HAZRAL MDS UD
IMPLE NUMBER: 50202473 IMPLE: T-3727
PAGE
7
:CD ORAL SCREEN
(CONTINUED) Clinical Signs (continued)
Hours 1.0 2.5 4.0
.Dosaaa Level
0.5 g/kg
Appeared normal 4
3
2
Diarrhea
1
2
3
Hypoactivity
0
0
0
Ataxia
0
0
0
Dyspnea
0
0
0
,Red ocular
discharge
0
0
0
Red-5tained face 0
0
0
Brown-stained anal area
0
0
0
Red-stained
genital region 0
a
0
Hypersensitivity
to touch
0
0
0
Piloorection
0
0
0
Thin appearance 0
0
0
Death
0
0
0
Appeared normal 4
3
4
Diarrhea Hypoactivity
1
2
1
0
0
0
AtaxiaI
0
0
a
Convu sions
0
0
0
Subconvulsive
jerking
0
a
0
Hypersensitivity
to touch
0
0
0
Piloerection
0
0
0
Brown-stained anal area
0
0
0
Red-stained
genital region 0
0
0
Red ocular
discharge
0
0
0
Red-5tained face 0
a
0
Thin appearance 0
0
0
Death
0
0
0
Days 1 2 3 4 5 6 7 8 9 10 11 12 13 14
Males
00 000 00 0- 44 3 000 0 0- 5 432 22 1 012100000- 10 00000 a- -
0010a000- 44 3 1 1 1 0 0- -
4432 22 10- 0 2 2 1 0 0 0* 0 - 00001 10a- 00000 1 10- 0010a 000- 01110011- -
Females
00000 0000 0 0 0 0 0
453 0000 00 0 0 0 0 0
55 54222 22
22
12
1
0 0012 222 2 3 3 2 2 1
0 000 a 00 1 1 0 0 0 0 0
00001 00 00 0 0 0 0 0
0000 000 00 2 2 1 2 1 00000000a 2 2 1 2 1
45 4444 44 4 4 3 2 2
0 1 1 0 0 0 0 0 0 .0 0 0 0 0
001 1111 11 0 0 0 0 0 3 3422 2222 3 3 2 2 1 00000 000 1 3 3 2 2 1 0 0 a 1 0 0 0 0 0 0 0 1 1* 0
*Animal died in p.m.
gab HAZLC%CN - w
LAsoPt=CRIES
AAAL:RICA. iPr-
Chemical & BloMedical Sciences Division
---
3301KINSMAN BLVD. a P.O.BOX 7545 MADISON,WISCONSIN 6370," PHONE (6082)41-4471 TLX 703956HAZRAL MDS UD
@MPLE NUMBER: 50202473 KMPLE: T-3727
PAR.E 8
:-'CDORAL SCREEN
(CONTINUED)
Clinical Signs (continued)
Hours 1.0 2.5 4.0
Days 1 2 3 4 5 6 7 "B 9 10 11 12 13 14
Do:5act Level - 2.00 g/kg
Appeared normal 3 2
2
Diarrhea
2
3
3
Hypoactivity
0
0
0
Ataxia
a
0
Brown-stained anal/
genital area 0
0
0
Death
0
0
0
Appeared normal 3
2
2
Diarrhea
2
3
3
Hypoactivity
0
0
0
Ataxi
0
0
0
Red-.satained face a
0
0
Dyspnea
0
0
0
Lacrimation
0
a
0
Yellow-stai-ned abdomen/anal/
. genital area 0
0
0
Death
0
0
0
Males 0 0 ---1 0 ---3 0 ---2 0 ----
2 0 -- -2 3 ----
Females 0 0 ------3 0. - - - - - - 4 0------4 0 ----- 3 0 ------1 0 ------2 0 -------
33* 02 -- --- -- -- -- -- -- - - - - -
Do5ace Level - 5.00 g/kg
Appeared normal 5
4
4
Diarrhea Hypoactivity
0
1
1
0
0
0
Ataxia
0
0
0
Brown-stained
anal region
0
.0
0
Death
0
0
0
Appeared normal 4
0
0
Diarrhea
a
4
4
Hypoactivity
1
1
1
Ataxia
0
0
0
Dark-stained no5e
and mouth
0
1
2
Death
0
0
a
*Two animals died in p.m.
Males 0 a ------- - - - - 2 @O - - - - - - - - - - - 3 0 -- -- --- - - - 3 0 ------- - - - - 2 0 ------- - - - - 2 3 ------- - - - - -
Females 0 0 - - - - - - -- - 3 0 ------- - 3 0 ------- - 3 0 ------- - 1 0 ------- - 2 3 ------- - -
OHAZLMZCN
L&A3CWt4MMRiES
AHAL:RICA. lf'S--
Chemical& BloMedicalSciencesDivision
3301KINSMAN BLVD.a P.O.BOX 7545 MADISON,WISCONSIN 53707 PHONE (6082)41-4471TLX 703956HAZRAL MDS UD
@MPLE NUMBER: 130202473 IMPLE: T-3727
PAGE
9
-.CD ORAL SCREEN
Dosage Level:
Animal
Number
Sax
C28605
m
C28574
m
C28604
m
C28547
m
C28414
m
C2e329
F
C28394
F
C28395
F
C28346
F
C28399
F
(CONTINUED)
PATHOLOGY 0.20 g/kg of body weight
Date Dosed: 03/26/85
Test Day Died Sacrificed
14
Necropsy Comments
Diffuse alopecia on ventral abdominal.region.
14
-
14
11
-
No visible lesions.
No visible lesions.
Red parinasal discharge; parinsum stained brown.
-
14
Diffuse alopecia an ventral abdominal region.
-
14
-
14
-
14-
-
14
-
14
No visible lesions. No visible lesions. No visible lesions. No visible lesions. No visible lesions.
HAZLEOCON LAEK3qM'OqieB ANAERICA, INM
Chemical& BloMedicalSciencesDivision
3301KINSMAN BLVD. 9 P.O.BOX 7545 MADISON. IISCONSIN53707 PHONE (6082)41-4471TLX 703956HAZRAL MDS UD
IMPLE NUMBER: 130202473 IMPLE*. T-3727
PAGE 10
-'CD ORAL SCREEN
Dosage Level:
Animal
Number
Sax
C29302
m
C29313
m
C2?575
m
C28418
m
C27576
m
C28529
F
C28534
F
C28537
F
C28532
F
C28533
F
(CONTINUED)
PATHOLOGY (continued) 0.50 g/kg of body weight
Date Dosed: 03/12/85
Test Day Died*Sacrificed
4
Necropsy Comments Periocular, parinasal., and parineal areas stained dark brown; lungs diffusely dark red.
2
Stomach - multiple, dark brown foci, up
to 3 mm in length, on glandular mucosa.
3
Dark red pariocular
discharge
(bilateral); parinsum - stainad dark
green; stomach - glandular mucosa
diffusely red, with multiple, dark
green foci, pinpoint up to 1 mm in
diameter, on nonglandular mucasa.
8
Stomach - raised@ tan areas, up to
2 x 2 x 1 mm, on nonglandular mucasa.
7
Parinsum/parianal area stained
dark brown; stomach - contains dark
brown material, glandular mucosa
diffusely red, with multiplep raised
areas, up to 1 mm in diameter, on
nonglandular mucosa;'small intestine
contains dark brown, mucoid material.
12
Stomach - dark brown areas, up to
x 5 mm, on glandular mucosa, with
raised, white areas, up to 1 x 3 x
2 mm, on nonglandular muco5a.
-
14
4
-
No visible le5ions.
Brown parinasal stain; stomach dark brown foci, up to 2 mm in diameter, on glandular mucasa.
-
14
13
-
No visible lesions.
Animal thin; stomach - multiple brown areas on gl-andular mucosa; -liver accentuated lobular pattern on all lobes.
HAZLMZCN
3301KINSMAN BLVD.
LA13CR=CYMES
AA@r--RICA.
P.O. BOX 7645 MADISON, WISCONSIN
53707
Chemical & BloMedicalSciencesDivision
PHONE (608)241-4471 TLX 703956 HAZRAL MDS UD
IMPLE NUMBER: 50202473
IMPLE*. T-3727
PAGE 11
:CD ORAL SCREEN
Dosage Level:
Animal
Number
Sax
C29326
m
C29328
m
C29330
m
C29335
m
C29331
m
C28664
F
C28497
F
C28695
F
C .28693 F
C28687
F
(CONTINUED)
PATHOLOGY (continued) 2.00 g/kg of body weight
Date Dosed: 03/06/85
Test Day
Died Sacrificed
1
1
2
2 2 I
1 2 2 1
Necrapsy Comments
Stomach
contains normal food and
tan granular material; small
intestine - filled.with tan/yellow,
mucoid semifluid.
Stomach - contains normal food and tan granular material; small intestine - filled with tan/yellow,
mucoid semifluid.
Stomach - glandular mucosa diffusely red; liver - accentuated lobular pattern.
Liver - accentuated lobular pattern.
No visible lesions.
Stomach - contains normal food and tan granular material; small intestine - filled with tan and clear,
mucoid semifluid.
No visible le5ions.
No visible lesions.
Liver - accentu-ated lobular pat.tern.
Liver - accentuated lobular pattern.
HAZLEZON
3301KINSMAN BLVD.
LMK3MOURIEB
AK4ERlf--A. W@C.
P.O.BOX' *45 MADISON, WISCONSIN
53707
Chemical& BloMedicalSciencesDivis@ion
PHONE (608)241-4471 TLX 703956 HAZRAL MDS UD
IMPLE NUMBER: 50202473 ;MPLE: T-3727
PAGE 12
."CD ORAL SCREEN
Dosage Level:
Animal
Number
Sax
C28545
M
C29314
m
C28198
M.
C29319
m
C28456
m
C28498
F
C28501
F
C28500
F
C28499
F
C28503
F
(CONTINUED)
PATHOLOGY 5.00 9/kg of body weight
Date Dosed: 03/01/85
Test Day Died Sacrificed
I 1 1
2 2
Necrapsy Comments
Perinsum - stained brown.
Perineum - stained brown.
Perineum - stained brown.
Red parinasal discharge.
Perineum
stained brown; red
pariocular discharge (bilateral);
red parinasal discharge-.
I
Parineum - stained brown; lungs
dark red and firm; thoracic cavity
contains a light tan granular
material.
1
Parineum - stained brown.
2
Perinsum - stained brown; red
perinasal discharge.
2
Perineum - stained brown; red
perinasal discharge.
2
Parineum - stained brown; red
parinasal discharge.
Deviations from the protoccli Some rats received a commercial laboratory food other than Purina Rodent Chow. During the study period the temperature of the animal room ranged from 21 to 25 degrees C. These deviations are not considered to have had an effect on.the validity of the study.
References: Organisation for Economic Cooperation and Development's Guidelines for Testing of Chemicals) Section 401, Acute Oral Toxic*ity,, adopted May 12, 1981.
HAZLOTON
LABCFWnX:41ES AN@ERICA. INC.
3301 KINSMAN BLVD. o P.O. BOX 7546 MADISON, WISCONSIN
53707
Chemical& BloMedicaSlciencesDivision
PHONE (608)241-4471 T')i-'703956 HAZRAL MDS UD
IMPLE NUMBER: 50202473 IMPLE: T-3727
PAGE 13
'CD SKIN IRRITATION
Objective: To determine the relative level of primary skin irritation/ corrosion of a test substance on rabbits under somioccluded conditions according to the Organisation of Economic Cooperation and Development's Guidelines for Testing Chemicals, Section 404, Acute Dermal Irritation/ Corrosion, adopted May 12,, 1981.
Test Material: T-3727
Physic al Description: Purity' and Stability:
Off-white waxy solid Sponsor has purity and stability determinations on file.
Test Animal: Young adult rabbits (approximately 14 weeike of age) of the New Zealand White strain were procured, maintained individually in 5creen-bottom cages in temperature- and humidity-controlled quarters, provided continuous access to Taklad Laboratory Rabbit Diet and water.,
and-held for an acclimation period of at least 7 days.
Three acclimated female animals, weighing from 2840 to 3112 g, were chosen at random for the test,,-treated,,and maintained during the observation period as specified for the acclimation period. Test animals were*identified by anima 1 number and corresponding ear tag. Approximately twenty-four hours *before treatment the hair was clipped
from the back of each animal.
Reason for Species Selection: Historically, the New Zealand White albino rabbit has been the animal of choice.for evaluating the effect of chemicals on the skin.
Preparation of Test Material: The sample was dosed as received.
Treatment: The test material was applied to the intact skin of each rabbit in the amount of 0.5 g per site and moistened with 0.9% saline. The treated area was covered with a 2.5 x 2.5-cm gauze patch secured with paper tape and overwrapped with Saran Wrap and Elastoplast.tape to provide a somiocclusive dressing. Collars were used to restrain the test animals for the 4-hour exposure period.
HAZLOZON
LABCXUUURIEB
ANAERICA. INC.
3301KINSMAN 81.1@0. 9 P.O.BOX 7645 MADISON, WISCONSIN
53707
Chemical& SloMedicalSciencesDivision
PHONE (608)241-4471 TLX 703966 HAZRAL MDS UD
IMPLE NUMBER: 150202473 IMPLE: T-3727
PAGE 14
.-'CDSKIN IRRITATION
(CONTINUED)
Observations: After the exposure period, the patches were removed. The test sites were wa*hed using lukewarm tap water and disposable paper towels. The test material was removed from the-test 5ites as thoroughly as possible without irritating the skin. Thirty minutes following removal of the test material, the degree of arythema and edema was read according to the Draize* technique. Subsequent examinations
were made at 24, 48 and 72 hours after patch removal.
Individual body weights were taken just prior to a.tudy initiation.
Pathology: At study termination all animals were authanatized and discarded.
*Draize., J. H.., "Appraisal of The Safety of Chemicals in Foods., Drugs and Cosmetics - Dermal Toxicity." Association of Food and Drug Officials
of the U.S., Topeka, Kansas, pp. 46-59 (1959).
HAZLEMON
LAEX3F44913RIES APAERICA. N%r-
3301KINSMAN BLVD. 9 P.O.BOX 7545 MADISON, WISCONSIN
63707
Chemical & BloMedicalSciencesDivision
PHONE ','082)41-4471 TLX 703956 HAZRAL MDS UD
IMPLE NUMBER: 50202473 IMPLE: T-3727
PAGE 15
'@CDSKIN IRRITATION
SUMMARY
(CONTINUED)
Test Animal: Albino Rabbits - New Zealand White Source: Hazleton Research Products., Inc., Denver PA Date Animals Received: 02/05/85
Temperature and Humidity of Animal Room: 20 - 22 Degrees C.; 40 - 44% Relative Humidity
Date Test Started: 03/01/85
Date Test Completed: 03/04/85
Animal Number
F07819 F07IB16 F07800
Mean
Individual Dermal Irritation Scores Test Material: T-3727
Erythema Score Hours
4 24 48 72.
Edema Score Hours
4 24 48 72
0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0
0.,o 0.0 0.0 0.0
0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0
0.0 0.0 0.0 0.0
0.0 0.0 0.0 0.0
0.0 0*.o 0.0 0.0
Primary Dermal Irritation Scores
Observation Period
3 Rabbit Mean
4 Hours:
0.0
24 Hours:
0.10
48 Hours:
0.0
72 Hours:
0.0
Results:
No dermal irritation was observed at any time during the study period.
Deviation from the protocol: The test material was moistened with 0.9% saline rather than deionized water as stated in the protocol. This deviation is not considered to have had an effect on the validity of
the study.
HAZLEMON
LABCFUUMIES
AKAERICA, INM
3301 KINSMAN BLVD. o P.O.BOX 7545 MADISON, WISCONSIN
53707
Chemical & BloMedicalSciencesDivision
PHONE (608)241-4471 TLX 703956 HAZRAL MDS UD
IMPLE NUMBER: 50202473
IMPLE: T-3727
PACE 16
7.CD SKIN IRRITATION
(CONTINUED)
References:
1. Organisation
for Economic Cooperation and Development'5
Guidelines
.for Testing of Chemicals, Section 404, Acute Dermal Irritation/
Corrosion, adopted May 12, 1981.
2. Draize, J.H., "Appraisal of the Safety of Chemicals in Foods,, Drugs., and Cosmetics - Dermal Toxicity",, Association of Food and Drug
Officials Of the U.S., Topeka, Kansas, pp. 46-59 (1959).
dab IWIWI-41
HAZLOTON
LPJBCWTAMMIEB
AK#IERICA. INC.
3301 KINSMAN BLVD. - P.O.BOX 7545 MADISON, WISCONSIN
Chemical & BloMedical Sciences Divliion
';3707 PHONE (608)241-4471 TLX 703966 HAZRAL MDS UD
IMPLE NUMBER: 50202473 IMPLE: T-3727
PAGE 17
7.CDEYE IRRITATION
Objective: To determine the level of ocular irritation produced following a single exposure of a test substance to one eye of albino rabbits accordingf to the Organisation for Economic Cooperation and Development's Guidelines for Testing Chemicals, Section 405, Acute Eye Irritation/
Corrosion, adopted May 12, 1981.
Test Material: T-3727 Physical Description: Purity and Stability:
Off-white waxy solid Sponsor has purity and stability determinations
on file.
Test animal: Young adult rabbits (approximately 14 weeks of age) of the New Zealand White strain were procured, maintained individually in screenbottom cages in temperature- and humidity-controlled quarters.,provided continuous access to Tekl-ad.LaboratoryRabbit Diet and!watori,apnd hold-
for an acclimation period of at least 7 days.
Three acclimated female animals, weighing from 2666 to 3BaO g. were chosen at random for the test. The animals' eyes were examined within 24 hours prior to test material administration using sodium fluorescein dye procedures. Only those animals with no sign of ocular injury or irritation were used. Test animals were identified by animal number
and corresponding ear tag.
Reason for Species Selection: The Now Zealand White albino rabbit is the animal of choice based upon its large orbit and nonpigmented iris.
Preparation of Test Material: The 5ample was dosed as received. A bulk density determination was niade to determine the weight equivalent of a 0.1 ml dose. Based upon the density determination..,an individual
dose of 0.09 g was weighed out for each animal.
Trea tment: Each rabbit received 0.09 g (0.1 ml weight equivalent) of the Ct,e':t material placed on the everted lower lid of one eya. with the tralateral eye serving as the untreated control. The'upper and lower
lids were gently hold together for one second to prevent los5 of material
and then released. The eyes of the rabbits remained unflu5hod.
HAZLMZON
LAEK3RArORIEB
ANOERICA. lKr-
3301KINSMAN BLVD. 9 P.O.BOX 7545 MADISON, WISCONSIN
53707
Chemical & BloMedical Sciences Division PHONE (608)241-4471 TLX 703956 HAZRAL MDS UD
IMPLE NUMBER: 50202473 IMPLE: T-3727
PAGE 18
:CD EYE IRRITATION
(CONTINUED)
Observations:
The treated eyes were observed for ocular irritation at
11 24, 48, 72 and 96 hours after treatment. At the 72-hour reading,
sodium fluorescein was used to aid in revealing possible corneal injury.
Irr'itation was graded and scored according to the Draize* technique.
Animals were weighed just prior to test material administration.
Pathology: At study termination all animals were outhanatized and discarded.
*Draize, J.H., "Appraisal of'the Safety of Chemicals in Foods, Drugs, and Cosmetics - Dermal Toxicity." Association of Food and Drug Officials
of the U.S., Topeka,*Kansas, pp. 49-51 (1959).
HAZLOTON
LAEKX4=URIeS
AT.AERICA. W%C.
Chemical& B!oMedicaSlciencesDivision
3301KINSMAN BLVD.o P.O.BOX 7545 Mt iiSON,WISCONSIN S3707 PHONE (6082)41-4471 TLX703956HAZRAL MDS UD
IMPLE NUMBER: 50202473 IMPLE: T-3727
PAGE 19
.'CDEYE IRRITATION
SUMMARY
(CONTINUED)
Test Animal: Albino rabbits - Now Zealand White Source: Hazleton Research Products.. Inc.,, Denver PA Date Animals Received: 02/05/85
Temperature and Humidity of Animal Room: 19 to 22 Degrees C.; 40 to 44% Relative Humidity
Test Material: T-3727 Date Test Started: 02/28/85
Date Test Completed: 03/04/85
PRIMARY EYE IRRITATION
OBSERVATION PERIOD
1 Hour: 24 Hours: 48 Hours: 72 Hours: 96 Hours:
SCORES*
.3 Rabbit Mean . 0.09 g (Unwashed)
7.0 3.7 3.0 2-3 0:0
The Primary Eye Irritation Score is the total eye irritation 5care for all the animals divided by the number of animals (3) at each
observation period.
Comments: No pain response (vocalization) was elicited from any animal following instillation of.the test material.
No corneal irritation was observed during the study.
(EHAZLC%CN
LAECP,=C)MES
ANAERICA. INC.
3301KINSMAN BLVD. o P.O.BOX.7545 MADISON, WISCONSIN
53707
Chemical& SlaMedicalSciencesDlvition PHONE (608)241-4471 TLX 703956 HAZRAL MDS UD
@IMPLE NUMBER: 50202473 IMPLE: T-3727
PAGE 20
'.CDEYE IRRITATION
Animal Number F07813
F07814
F07815
Observation Period
1 Hour 24 Hours 48 Hours 72 Hours 96 Hours
1 Hour .24 Hours .48 Hours
72 Hours 96 Hours
1 Hour 24 Hours 48 Hours 72 Hours 96 Hours
(CONTINUED)
Table 1 Individual Eye Irritation Scores
Cornea
A
B
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
Score AXBX5
0 0 0 0 0
0 0 o 0 0
0 0 0 0 0
Iris Score
A
A x5
1
5
1
93
1
5
1
9
0.
0
0
0
0
0
6@
o.
0
Q
0
0
a
0
0
0
0
0
0
0
0
Conjunct.ivae Score
A
B
c (A+B+C)2
1
1
1
6
1
1
0
4
1
1
0
4
1
0
0
2
0
0
0
1
1
1
6
1
0
a
2
:o , -o
0
o;
0
0
0
0
0
0
0
1
1
0
4
0
0
0
0
a
0
0
0
0
0
0
a
0
0
a
0
Table 2 Sodium Fluorescein Examination
Animal Number
F07813 F07814 F07815
Observation Period
Pro-initiation
72 Hours
NEG
NEG
NEG
NEG
NEG
NEG
NEG - No stain retention POS - Positive stain retention (area of cornea involved).
References:
1. Organisation for Economic Cooperation and Development'5 Guidelines for Testing Chemicals, Section 405, Acute Eye Irritation/Corrosion, adopted May 12, 1981.
2. Draize J.H... "Appraisal of the Safety of Chemicals in Foods, Drugs,, and Cosmetics - Dermal Toxicity",, Association of Food and Drug Official Officials of the United States, Topeka, Kansas, pp. 49-51 (1959).
PAGE 21
QUALITY ASSURANCE STATEMENT Study No. 50202473
The report as herein attached for the above-mentioned study has been reviewed
by the assigned Quality Assurance Unit of Hazleton laboratories America, Inc. in accordance with the Good Laboratory Practice Regulations as set forth in 21 CFR 58.35 (b) (6) (7). 'Ithas been found to accurately identify and/or describe the authorized methods and'standard operating procedures followed in
the conduct of the study and that the reported data accurately reflect the raw data of the laboratory study. Furthermore, the Quality Assurance Unit has conducted the following inspections of the testing facilities @tilized in the conduct of this study and has submitted written reports of said inspections to
the study director-and/or management.
Date of Inspection
Type of Inspection
Acute Oral Toxicity Study in Rate
2/25-26/85 3/26/85 5/01/a5
Process Audit Process Audit Report Review
Primary Dermal Irritation Study in Rabbits
2/25-26/85 5/01/85
Process Audit Report Review
Primary Eye Irritation Study -inRabbits
2/25-26/85 5/01/85
Process Audit Report Review
Date Issued to Management
2/26/85 3/26/85 5/01/85
2/26/85 5/01/85
2/26/85 5/01/85
zo
Diana-E. Skalitzky Inspector, Quality Assurance Unit
Date
ACUTE ORAL TOXICITP(LD50) RECORD
Test Haterial T-.5la7
CIO
iioo.03OIL-
RT No.
Bulk Density
(S/nl) Species Fasti@4: Dote
Source 90.r(&n
Date Received
Tim 2.-'3g0.P-Tech. CK -Room
Doisafte 0,20 C&L Dogs Volum
Sex
Animal No./Ear TallNo.e
Protasted lgdv Wetaht (a
FLFasted Body WeiRlit (R)
Done m ]!s5s-,cai
79Q
73 71
0
Tech. DLYIT s
Actual Dose
PDraoy" 7 Body Weight (g) ADDDcaa8ayty"t14 B-ody Welgbt t
37
3.1
Verified b
Dosagg
r),I
/k
Dose Volume 10.0- (al/kg)
Be I
Animal No./Ur TaftNo
39q 140;k 95,
Prefaated Body
rFFasted Body wW lght (R
-
q ,.a- 111 .700
APID,racctual Dose ( 1
.0
Day 7 Body Weight (g)
Day14 BodyWeight(s)
;119
Teche Are
ga OIL C2.
hones v r tied hy
3-al
Sf '44t
Dose Level
lioura U-o4 urg4
.3
lkd Teclin,i@)rin
HORTALITY (NO. DIED/NO. DOSED)
Dav
2 -3
4
DR Ina
60
016
ol 0
L
L IrL-
6
-itLCk
9 io BJL
Date
7- 43
NA - Hot Appltcphle
Reviewed by
t
- Waage calculated, but not adminiiitPrIB4
lintifiaeltiimnl returned to stock
ACUTE ORAL TOXICITY (LP.50)RECORD
Test Haterial
'T-A7.Q-7
v4phLcle.
C-ne-bi C)IL-
RT No
Bulk Density IUA
(9/141) Species
Source "O@fkn-n
Date Receive
raisted: Dote
Tim 3.'oao.vh.Tech.latm --R.oom
Donag@
52
Does Volwn io,D (al k
Sex Animal No./*Ear Tait-No.Ca 3Da 93L3
Dose as i.,z55,o,.
lig
Tech 3-i;t
Prefasted 12dy
Fasted Body.Weigilt (g)
7
-ia
Actual Dose (al
Day 7 Body Weight (s)
Demb Palo -110 3*iq-lw E@.T-
Q34,
Day 14 Body Welitht (&I
Doses Verit:Le Oil
Dosage C).Sb
Dose Volum
(jr/kg)
wee It" 30 0.-VA-
_AnlilmaNlo./gar Tax No
7
Prefasted BodX Wet bt
A
Fasted Body VOiRbt (R)
3(o
010
Teidh,
Actual Dose ( ut Day 7 Body Wel@lit
Day 14 Body Weight (g) s -.2-qs
all=
lftedb3i
HORTALITY(NO.DIED/NO* DOSED)
Doi3e
IlOuvB
s
Lev6LDeovle3le
04
1 liP2 3
4_
5
6
a
21&L
8
9 10
-q)4&1 KaL .ofI 'r
D,
Tecitar@ia@'a
-P <-. @QL.-Co
-Date ige6S
ir3
NA - Hot Applica0le
- I)osagecalculated but not odataistered llnilqen(iliimnlretiirnedto stock
(A C44
c9 -31
Reviewed by #irC
Date D4
Test Haterial
3 227
ACUTE ORAL TO@XICIT@P.(LD 50 RECORD -Vehicle- et)FAl
RT H
Bulk Density
tjft -(B/Ml). Species e.&-+ Source A&-c-I*-n
Date Receiv
F&pted: pfte
Time P?:DOPM Tech. e-& Room
DOBaR6 Dose Volu-me lt).O $ex Animal No./gar TaitNo
a(p 9330
Doise me 33-1- q 3,2g q3;1,7 1.31_1
%,11 3 -
Pretasted Bod!i Wein4 Fasted Body Weight
L AA ctual Dose (al
D"ay 7 Body Weight (g) PDra" Dacay 14 Body Welgbt
7 1(603
OF
on
70 al
.i.l
3-t-ar
bir
;t47 ;L. i.s. In$ .4-6-is 3
D-poes Ver tied bvi IYI
Dosage
C)Q (R/kR)
Dose voltme Animal No./gar-Tait W.Ca
i(o(,3
Prefasted Body Weiglit(it) Fasted Body Weight (it)
@51 ;t33 .253
-Actual Dose ( nt Day 7 BodZ Weight (g)
Day 14 Body Weight KtWom
ia
s 3-1-dis ion_I& (,*A
MORTALIT; (NO. DIED/NO. DOSED)-
D4s Dosea
liours
Dav
Level Levce)'l
o"D O"r4o Wi1t
23
4
5
6
7 1-8
9
am Una IM AS UL
J?JL
UL
km
Tacit.
---10 ].1 gamma
Date
II
"A - Not Appi@cable
owrjy)i erro,-a--i-sstepvjPlripd by
Dg
- I)osagecalculated but not administered@ -T,.d-"+e-.0i01-14cvr.1,l@kerr4&u4EAvvs@l
.1 ret roed to stock
A
Vk&G -C Vol vcl
1,.A;"f 214 -j
Test Hategla]L Bulk Density
UbBaRS Does volume l(@'D
Sax Animal No./Bar TO
Prefasted 12ay weigh Fasted Body Weight Actual Does (al Day 7 Body_weight
Day 14 tioditweight tal
0
ACUTE ORAL TOXICITY (Lp.50) RECORD
vehicleer3nr o i
Source
RT No pate Receive
rasteds Upto
as Tim
7314 opt
Time S-02d6AA Tscb- -6#"t-Room -
AW-AW
Tech. Date
1311
q?
ll.()
&,Mon
Vpelf I-A bvi
Dosage
,T,no
(a/k
Dose Volume lo.n
Animal No./ga-r T4K
Pretasted Body VoiliLit Fasted Body WeilLht
Actual Done ('81 Day 7 Body WeiRht-(g
idyWeight (a)
Dose
Hours
Level
o-4
1
no,taoroe- '6ezr9D#e 00
Ila #I
Tech.
.731
-ILi
-107 :1.25 q
2.,aL7.4
S;;j@"
iN@ 3 g Ve itted by
MORTALITY (no. DIED/NO. DOSED)
Dav 2
3
4
-5- 6
7
9
ML
91<
Teclialeenn
Date lql-T- 3/1
NA - Hot Applicable
- I)OOSsecalculat:d
d
llnistiaclillinalr titbruote4ntoo@ satdo*ctknioter
Reviewed by
Da
A,iV&
UT me. io@2i3.2A7I
lr#st OTMIAL. -r
fffuov TRUXI
su
OWAalt LIIVBL
STUOV PAV Am---NATS'
ql 71
ti'
rj
11112 Li iq
-jeClan (.1
Ct,i1i4
IA DATR
\00'- Sills rt*gdlbt
31 J)DIED lt4
Nu - not IIA - Not Appiic&Uke
Sf
ATUOV TITLgs
UT NO. TIMT HATUIAL
7-3
-r-57J-7
su
T DoAas IAVGL
lllnov
simmora "a AlPfEAPfjlW Om U
e
0
I IQ3
Aif Aumi-
pe 06 Al
V)6
121103lq
&001
3 PATR
also remodat slan rcesent, slighc
st obselfvttiiondotif,oil
pot ftlov
Us - Not iguilause "A - flat Appoleahle
o q-,D-4!5 et,-
EMV T&ILKS
OIT10.S020.2171
-tautimirmiAL r--vF..2'7
1
"SAGIS exl*SL
3 siumaxo AIPI?NANI
(L A
I-ivttd
PA #-if,'
to It12 t3llq
730
tic
+0@* C-
Ail4kvv/
09AIN lemiCIAN
$one
rfamout.
sign Vroments
slight
m - "at
MA - Hot
otwi4leat
App I I ca4i a
VMV TITIAs
me.@,VO,2i73.2Y
-rimIv"TIMIALZ--'A
au
TA 00"Ga LOIL
sumv PAY 3
un
IP A*
file
A4o@*ic-
lii!Alr Tti:mmgcAM
IDATI,
'130
$INS read"& as alga rcdoango $fight
INV
yl1t',/7
a
Ng - Mat ltwAent OU - Nat Applicable
one C
lr\
VI -.o
000ol
.oo oor
1000011,
0000,
-.00
84
UUDV TITIAI
TIMT,u&TatAL r-,A
su
TA DN"a IAVBL
STioV PAV AIPIPEARZO
(0
.30 31
e
1133' 1'4
id oe-
-171!A-Ti OATS
$INS sign rcesents slight
NK - Not avident MA - Hot Applicable
-4
r
7-1
Eta--,
<-- Crh CrW
CA
0#100
'4o0o,001-I I FI-J @oe
II
a T u 0. so o gil-
TIMT amir=iAL r- P-2-7 LWBL T
PAV
SrAwonim PAT#
A r AltisNo]" 5@wicj @ace-
eA ox^;44.s
q
XV//
ql4
4 '17
v./
Tl!io.m IAN
also
al sign redgent.
slight
SP
UK Nor
MA Hot Applic&Ule
"mv
OITme.90;2tl.2Y.7
Iran iavni&L r-
TinAt
q s-nov PAV
.
simvmu" un
-EM
octti
"$AM
su
SL ..0!.J@ IAW
q y/9
,WAN irmiciali
"TR
D /3;
Sligo rfandat
21$A rcogent, al ltht
Not swidest
Applicable
awv N:wowim
IIIA@O I"
I-IIA,
90"109 LBVRL
Y/A A 3
2 13liq
ql.q
1/9
34; PATK
some reason& also Firement. slight
Y/7
UK - Mat Swident MA - Not Applicable
STMOV TITISs
OITNO.SOAoA-I-ZlL-
TIMT HATIMIAL :E- %772-7
Ali
kA P:rpEm, ail
Le
149
aim
onsm OAVBL A.-go,& 1-ilik
liq
Adil A
LV/ loIoA
4
IIVATU
PATIE
till
L-4.
)/13
Some ressamt ol Alan reasent. al litht
0
Nu - Hat KV14009 MA - Hot Applicalsto
ST mo.5oaoA47 1riMMTTIMIAL
STUOV
glUOVDAV
saF',E:@
ck.&&-
OA
I:UI20
N+axic,
V
on"as IZVSL
ou We
iq
L)EATU
"Tit
Sf
3/la go-go reagent
al lose rfogauto slight
us - Not gwident liA - liat Applicable
ami
liTNo.SoaoAq*7,&.
TJWT H&TOIAL
Tglr$Ai
STIOV PAV
so3wim.0
ArfrAsis WOUL
a_
P&A
jA
sa "SAGS UVBL -oisna@@
10 11 12 13 1
lti!ATd' Ti-(:iwlclAm
IIATit
lit
silrltsemeat
sign froments ltht
Not liatAloplicolois
itTmi.
TimirISTIMIAL
in
to 12 13 iq
IIATR
son* $is*
slight
Na - Not av 140*t NA - Not Applicable
UT NO. Scjk;ID,?q7 TggT BATIMIAL -r-A"?
L3
su ONAGS izval
a ql IIQ 3 5 (P
"To[
IOU 49,,wtri,-
i4:W
al - sign reagents slight
Hit - NOR: ltv14001
MA - Hot Applicalois
PRIM&RY SKIN IRRITATION SCORING SCALE
(1) Erythema and Eachar Formation
No arythem 0
Very slight arythems (barely perceptible)
1
2
Well-defined erythosa
Moderate to severe erythema
3
Severe arythema (beet redness) to slight eschar
formation (injuries in depth)
4
Highest possible arythema score 4
(2)Edema Formation
No edema 0
Very slight edema (bartly.pareeptible)
Slight edema (edges of area well-defined by
definite raising)
2
Moderate edtm (raised approximately I on)
3
Severe edema (raised more than 1 m and extending
beyond area of exposure)
4
Highest possible edema score
4
. Date Aulm.le secolve4i
maid Abkl"lo Ciip"ils
skill trspd4sti4ang
4..16L
niVAil DUVAL IMITATION.WMV
vehicles ma;ipff-AIFZI tiif-h 0,111.0
SourcesUA-Lit4on I&A(rAoli-b
Tech$
lattlatodbys Reviewedbys
NIA NuaWto
liesults
Roommumbe
CAL
li..116.a FO
7@4? lgf
"Y Waigigt,(A)
0
*0
1"r
--49116
I*cbmiclna
by
Memo 12 lluut a
theme
7 I)ay lkuly lteigiol (a)
MA A rousibla muctutic area.
clic
govlewed bye
!$U
"to$
Cl)
(A) 92WA= dQV*
demdtr CWoa go" doM takle
or aff=e aea. ""I" of U-18 CLOW13, viame
3astUr"
trwwluamt w@t detalu ae iris
Nagar dloswame
2*
sugw27. obsoured -detalu of iris TIAUGO SLM of PUP" 3*
is
ope4m, ifts uwiam*
ca) An& it a=u ALULU, am 48mmw Car l@)o b" not,um wmtw two @ quwtw, but low than Ulf
W,@tw than wIt, b" low than thrm quertwo L.MSW than UK 4owtAn, up to vb*Wuma
Az 2 x 5
Tota nmxgum so
(2)
(A) ZRIM
norml
ftlow obwe pami. cmw or au at tbom
or =Ma"Un
an.raw"= to ugut, b.mwrmge.
'24 ne. aLta ow
vow
OW"t)
U48 au
4r
To"&
10*
(3) anamium
to papowel awwouvm
MIT)
CA) - am=
Cru'ws
V46mas awmi Tess*U docialtOL7 udgeted abo" WOOL
lbm dltf=wt ""W WIN=* r". -inuftem, "@"
2*
easily DLNWELUO
. ....
nfr@ b@dY red
(3) aMM10A
So MMLLUIS --
joW =WaUg abom mwml CisOLudm ALOUtatift
obvism @114
%tta partial eve@Mm at U"
3"U in With Luis ~
MU aloood
&AdLift IIL%BLt" aboutMU dAM"
aloo"
cc) ALUM=
ft dlaebwp AmW =want dUTOrMt
(dow ML I=Lu" Mau
aboa.vw in umm autbw of nor=L odmis)
or the U" 2
U"
tbo LUig and bal"g and oon"dersue.
usobwv Witb OBLaignin of
3
wm around ow "a
3owe (A o 3 # C) x 2
Toga VANd@m a 20
ran total sowo for the S" is the @ of all wo-se swed@a.iris, ad omjmativ"-
iadic&te.Poolti'we bffect.
obtain" -forthe
imary Eye T=itacion Test initialSodium Fluorasr-ain Ex= =d Ard=l
Bod-yWeights
Test Cm*ound
RT No.
<I-7
PH Result Doze
el
Data Art4msl[2 RaCfdV*d
e-rgiLa4;er*)
Room No. Dosed By Revieved BT
Data
to ----r-
sot=*H:azle@nlyRted,-n-rc-k
Animal No.
VOC&Liz
'Initialticm Sam. Sp* ftllaw:Ln
ftoecailal an@@t
Animal ]BftWody izhts (s)
iac:Lon
Da7 7
Day 14
FO- -71 a-
(a(-o
0-7
tien)
Da7 Zi
RZCOIMED
DATE
I2sic:zUSEZ)
Sodium Fluarescsin Exazination
NEG - Negative
POS a Posit:Lv*
o-f
IW
NA - Not Applicable
y - yes
N m NO
./-.'4'TeCsctapound.4 Test Eye ?N @o,@@-
Washed
@rimary Eye Irritation Test Observations
itTNo. rio0,2o7Y-7,3
Group
Seconds following instillation of test
material, the teks3tp0t7,0wfaosr was d With
N)A- al of
seconds
Unwashed
inal No./
rn"lL"ious ar Tax No.
cestioonf
No ail
Read
oa=vATiow Pmaoo:
@NnLkr-
7813-- -73r/(4 7RIS'-@
cular- Structure rnaceruII.rLu1-alTt"MaALrIou-STt-r0o0%;pacity
Area
Iris r.oujunctiv" -
Redness
Ch@asis Dischar"
edium Vluarasetin gxm
chnician
scor ad Bv
ta t*
Aa aa Ca D Inj HU
POS
Pm=lent Discharge Clear Discharge Petits amorrhate slauching lujo--ted Negative. Positive
za 7a C-
RIA
cormal lpithelial Damage, Corneal Epithelial Damage, Cormal Zpithelial Damage, ftume Corneal neawascularization Not Applicable
Peeling Piling Pitting
Reviewed By:
Date:
Eye Irritation Score:
(1311A)
l,@-N-4Test Compound
Test Eye
Washed
@,rimryEye irritationTest Observations
:3721
Group
PiA IT NO.
Seconds followinginstillation of test
mterial, the test *70 Was wasbpd with
N) A al of
or
seconds
V"O'
*7 2 Unwashtd
rFnt"3 imal No./ r Tax No. tiIoLnoefsions LuaT,a
FO-
OBSUVATT I
'7913 -7XIq
ail
Read().
-laSrtructure a Opacity
Area
ris
ALiLrs
7x/s--N
nu(
v," Ud@ness
Chmosis
Discharte
odium IFluorescein gxw
chon.icdian
corded By
A ftrutent Discharge 3 clear Discharge C Petite nomorrhate D Blanching INJ Injected MG Negative POS n Positive
Reviewd By:
.........
3/1
g Corneal gpithelial D=eage, Peeling IP Corneal Epithelial Damtet Piling r, coru"i zpicholial Damage, Pitting 3 Pannas I Cormal Neovascularization RA Not Applicable
Date:
zy* irritation Score: S.-75A-*
(1311A)
primary Eye Irritation Test Observations
Test Compound
Tesc Eye
0%amk
t-13Ot W"bed
Group
RT No.
N)A
seconds following instillationof test
material, the test eye was Was W2.th
N)A@alof -kit; for
seconds
Umashad
imal No./ ar Tax No. ocation of
roaal Lesions
Iaci.14
OBSERVATL
FO 7813--
!m%aims
cular Structure
rwa - 22&Cit-f
Are&
Pzuoo-. Wnur -,Fg s-
ris junativae-
Redness
h
1)
Choweis Discharte odium Fluorescein zxm
r)
A@.4
A)A
schnician
*corded1"B1v
or"d tacte..
3
Aa 3a Ca D I.NJ MG ?93
fttulout Discharge
Clear Discharge
Petite Remarrbag* stambing Injected Negative Positive
(DIN4oij,#VDr-S-Z-OIJVCorn"l gpitholial Danage,*Peeling I a Corneal Zpithelial Damage, Piling a a Corneal spithelial Dasage, Pitting B - Pamus I - Cormal Neavascularixation NA a not Applicable
Reviewed By:
Date:
Eye Irritation Scare: '3 C) (1311A)
,ntest Compoun d
Test 2 y*
e%
Ot Washed
is& ar Tax No.
ocationof I Lesions
Primary Eye irritation Test Observations
RT NO. riozo.7
Group.
JN-.Jrl
Seconds following instillationof test
mcarial,.the test ye 0 wasbpd with
of
f r K3t-t seconds
V/00' Unwashed
7913 OBSUVATI
PMAOD.- r @N^ar
7>?/S'm@
cular
Structure
mriurmais'aTtaar'3ria-S-Nto0oepri2citt,-7
Area
Iris
r,onjunativae-
Redness
Cheamis
Di
Sodium Moreseein-S.
Ai.
chnician
corded
19
MMMW,4
A a Purulent Discharge A a Clear Discharge
C a Patiia goorrhage D - sl&waing. INJ - Injected no a Negative
PCs a tositive
r)
0r,
Inn
E a Corneal Epitholial D@aget Peeling I a Corneal EpithelialDamg$t Piling a a Corneal EpithelialD@ge, Pitting a a Panwa I a covmal ileovascularixation MA a Not Applicable
Reviewd By:
-st.2. Date:
Eye irritation Scare: 2-S s%z, (1311A)
4'"".IsestCompound Test 270 t,3Pt. Washed
Primary tye Irritation Test Obp,..,vations
07'3:7-2-7
Group
RT No. S10:20;) (-173 QA
Seconds following instillationof test
mterial, the test eye we vashpd with
QA alof . U
fa: OM saccmds
Unwashed
I N0.1
r TaitNo.
tionof
rueal Lesions
a@aiiIl l-
- sead
et&likr structure
a 22&city
Area
Iris
7813
0
0 0
033MEnMIVAT=nl PMOD:., 0
'71rfq 7R/5'
u
0
0 0
\-,nLiLer,
Radmso
I)ischar" 0
Sodium Yluor*scain-Ezaii Ala
chnician
@carded Bv 9 st57
A a ftzulent Discharge a a Clear Discharge C a Petita Hemorrhage D a slambins Inj a Injected NU o Negative PCs a Positive
Reviewd By:
n
I Altq--
c P-
I Corneal Epithelial Dmage, Peeling Ir corneal Epithelial Da@get piling 0 Cormml Epithelial Dmage, Pitting 9.0 Paimas I a Cormal Neovascularixation NA a Not Applirable
Sk-t, Date:
Eye irritation Score: !!@L--, (1311A)
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HAZLE"CCNLABORATORIES
3301 KINSMAN BLVD. P.O. BOX 7545
AMERICA, INC.
MADISON, Wi 53707
(608)241-4471
TLX 7bM
HAZRAL MDS UD
February 25, 1985
Dallas D. Zimmerman, Manager, Toxicology 3M Center St. Paul MN 55144
PhD Services
International
Dear Dallas
Enclosed please find two copies each of the following protocols for sample T-3727, HLA No. 50202473:
Protocol No.
Study
TP-2069 TP-2072 Tp-2071
Acute Oral Toxicity Study in Rats Primary Eye Irritation Study in Rabbits Primary Dermal Irritation Study in Rabbits
These studies will be conducted in accordance with the OECD testing guidelines and GLP regulations.
Please sign all cop@ies, retain one set for your file, and return the others to me. We can initiate these studies upon your verbal authorization.
Should you have any questions, please feel free to call.
Sincerely
Steven M. Glaza Study Director Acute Toxicology
SMG/mvh Enclosures
Chemical & BioMedical Sciences Division
HAZLE"CDNLABORATORIES 3301 KINSMAN BLVD. - P.O.BOX 7545
AMERICA, INC. MADISON, Wi 53707
(608)241-4471
TLX 703956 HAZRAL MDS UD
PROTOCOL TP2071
Primary Dermal Irritation Study in Rabbilts (OECD Guidelines)
Study -No. 50202473
for
3M St. Paul, Minnesota
by Hazleton Laboratories America, Inc.
Life Sciences Division 3301 Kinsman Boulevard Madison, Wisconsin 53704
February 25, 1985 1985, Hazleton Laboratories America, Inc.
Chemical & BioMedical Sciences Division
PROTOCOL TP2071
Primary Dermal irritation Study in Rabbits (OECD Guidelines)
Study No. Study Location
50202473
Hazleton Laboratories America, Inc. Life Sciences Division 3301 Kinsman Boulevard Had*Lson,.Wisconsin 53704
Test Material
T-3727
Sponsor's Representative Study Director
Dallas D. Zimmerman, PhD Steven M. Glaza
Pi!oposed Timetable Starting Date Completion Date Final Report Date
W eek of February 25, 1985 Week of February 25, 1985
Week of April 1, 1985
OBJECTIVE The objective of this study is to determine the relative level of primary
on rabbits under aemioccluded conditions.. skin irritation of a test material All aspects of this study will conform to the Organisation for Economic
Cooperation and Development's Guidelines for Testing Chemicals, Section 404, 1 and the@U.S. Food
Acute Dermal irritation/Corrosion, Adopted May.12,.1981
and Drug Administration's Good Laboratory Practice Regulations for Nonclinical
2 Laboratory Studies.
All procedures will be done according to.Hazleton
Laboratories America, Inc. (HLA) Standard Operating Procedures (SOPS)
referenced in this protocol.
TEST MATERIAL
Test material: Physical Description:. Purity and Stability:
T-3727.
Off-white solid. Sponsor has purity and stability determinations on file.
Storage Conditions: Test Material Retention:
Store at room temperature.
Any unused test material will be discarded 30 days after issuance of the final report.
Safety Precautions:
Laboratory personnel will take the normal necessary precautions in handling a substance of unknown toxicity. Laboratory clothing, latex gloves, safety glasses, and a particle mask approved for, toxic dusts must be worn.
TEST SYSTEM Test Animal
Young adult albino rabbits of either sex of the New Zealand White strain, approximately 14 weeks of age, will be obtained from Hazleton Research
2
Products Inc., Denver, Pennsylvania. An adequate number of*extra animals will be purchased so that no animal in obviously poor health is placed on test. Historically, the New Zealand White albino rabbit has been the animal of choice for evaluating the effect of chemicals on the skin.
Acclimation Upon receipt, the animals will be taken to a designated animal room where
they will be acclimated for at least 1 week before being placed on test (OP-GENB 36). During acclimation, the animals will be exam@Lned for clinical abnormalities indicative of health problems (e.g., diarrhea, ectoparasites, rough hair coat, nasal or ocular discharge, evidence of injury, etc.). Any animals regarded as unsuitable for study purposes because of poor physical condition will not be released from acclimation and the reason(s) will be documented.
Identification Each animal in the study will be assigned a permanent identification
number and will be identified with a metal ear tag (OP-GENB 24). All data collected from an animal will be recorded and filed under its identification number.
Housing and Maintenance The following environmental conditions will be maintained in the animal
room used for this study (OP-TARC 230).
0
Temperature: 21*C +2@
0
Relative humidity: 50% +20%
0
Air change: At least 10 changes an hour of 'filtered 100% outsiae air
0
Light cycle: 12 hours light/12 hours dark
3
Temperature and humidity will be monitored throughout the study. Variations from prescribed environmental conditions will be documented.
Animal husbandry and housing at HLA comply with standards outlined in the "Guide for the Care and Use of Laboratory Animals.g,3 Care will be taken to ensure that the animals are not disturbed for reasons other than data collection and routine maintenance. The animals will be housed individually in screen!--bottosmtainless 'steelcages (heavy gauge) held on racks, with absorbent pan liners in the urine- and feces-collectingpans. Pan liners will be changed at least three times each week.
Feed and water will be provided ad libitum. The diet will be Teklad Laboratory Rabbit Diet. No contaminants are expected to be present in the feed or water which would interfere and affect the results of the study.
Stud Design Three rabbits will be selected at random based upon health and a body
weight of 2.0-3.5 kg. Each animal will serve as its own control.
PROCEDURES Preparation and Administration of Test Material
Twenty-fourhours prior to test material administration, the hair will be*. clipped from the back and flanks of each animal. The treatment sites will be inspected for interferinglesions, irritation, or defects that would preclude the use of any of the animals.
2 The test material will be applied to the test area (approximately 6 cm on each rabbit, in the amount of 6.5 g and will be moistened with deionized water. The treated area will be covered with a 2.5-cm x 2.5-cm gauze patch
4
secured with paper tape and loosely overwrapped with Saran-Wrap and Elastoplast tape to provide a semiocclusive dressing. Collars will be used to restrain the animals during.the 4-hour exposure period.
Reason for Route of Administration 4
Historically, the route of choice based on the method of Draize.
Observations After the 4 hours of exposure the patches and'the test material will be
removed as thoroughly as possible using water or an appropriate solvent without irritating the skin. Thirty minutes'after removing the patches, the degree of erythema'and.edema will be recorded according to the Draize Technique (Attachment 1). Subsequent readings will be taken at 24, 48, and 72 hours after patch removal. Further observations may be recorded, as necessary, to establish revei@sibility.if irritation is increasing in severity at the 72-hour examination period, observations will be repeated at 96 hours and at 7 and 14 days, if applicable.
Body weights will be taken just prior to test material administration and at weekly intervals during the study. Observations and body weights will be recorded in the study notebook.
Pathology All animals, whether dying on test or sacrificed at study termination,
will be discarded.
Re por t The final report will present a description of the test materials a
description of the test system, dates of study initiation and termination, a tabulation of irritation data, and a description of any toxic effects other than dermal irritation.
Maintenance of Raw Data and Records Original data or copies thereo,fwill be available at RIA to facilitate
auditing the study during its progress and prior t-oacceptance of the final' report. When the final report is completed, all original paper data, as well as the final report, will be retained in the archives of HLA, Madison, Wisconsin (OP-GEN 44).
REFERENCES
1. "Acute Dermal Irritation/Corrosion", OECD Guidelines for Testing Chemicals, Section 404, May 12, 1981.
2. 21 CFR 58 3. DHEW Publication!3 No. (NIH) 78-i3 (1978). 4. Draize, J. H., "Dermal Toxicity," Appraisal of the Safety of Chemicals in
Foods, Drugs, and Cosmetics, Asso@Lati@on of F-o-odand Drug Officials of the U.S., Topeka, Kansas, pp. 46-59 (1959).
6
PROTOCOL APPROVAL
Dallas D. Zimmer
PhD
Sponsor's Rep@Tesentative
3M
Steven M. Glaza Study Director Group leader, Acute Toxicology Hazleton Laboratories America,. Inc.
(10 69S/j g)
Date Date
7 ATTACHMENT I
PRIMARY SKIN IRRITATION SCORING SCALE
1. Erythema and Eschar Formation
0
No erythema
1
Very slight erythema (barely perceptible)
2
Well-defined erythema
3
Moderate to severe erythema
Severe erythema (beet redness) to slight eschar formation
4 (injuriesin depth)
4 Highest possible erythema score
2. Edema Formation
0
No edema
1
Very slight edema (barely perceptible)
Slight edema (edges of area well-defined by definite raising)
2
3
Moderate edema (raised approximately 1 mm)
Severe edema (raised more than 1 mm and extending beyond 4
area of exposure)
4 Highest possible edema score
HAZLC%CNLABORATORIES
3301 KINSMAN BLVD.
P.O. BOX 7545
AMERICA, INC.
MADISON, Wi 53707
(o'082)41-4471
TLX 7039% HAZRAL MDS UD
PROTOCOL TP2069
Acute Oral Toxicity Study in Ratt'
(OECD Guidelines)
FEr2,
7 lqt,-
Study No. 50202473
for
3M
St. Paul, Minnesota
by
Hazleton Laboratories America, Life Sciences Division 3301 Kinsman Boulevard
Madison, Wisconsin 53704
Inc..
February 25, 1985 1985, Hazleton Laboratoiies Americas Inc.
Chemical & BioMedical Sciences Division
PROTOCOL TP2069
Acute oral Toxicity Study in Rats (OECD Guidelines)
Study No.: Study location:
Test material: Sponsor's Representative: Study Director: Proposed Timetable
Starting Date: 'Completion Date: Final Report Date:
50202473
Hazleton Laboratories America, Life Sciences Division 3301 Kinsman Boulevard Madison, Wisconsin 53704
Inc.
T-3727 Dallas D. Zimerman, PhD
Steven M. Glaza
Week of February 25, 1985 Week of March 11, 1985 Week of April 1, i985
OBJECTIVES
To determine the acute oral toxicity produced when the test material is
administeredby the oral route (gavage) to rats. All aspects of this study
will conform to the Organisation for Economic Cooperation and Development's 1
Guidelines for Testing of Chemicals, Section 401, adopted May 12, 1981 and
the U.S. Food and Drug Administration's Good Laboratory Practice Regulations
2 for Nonclinical Laboratory Studies.
All procedures will be done according
to Hazleton Laboratories America, Inc. (HLA) Standard Operating Procedures
(SOPS) referenced in this protocol.
Test Material: Physical Description: Purity and Stability: Storage Conditions: Test Material Retention:
Safety Precautions:
TEST MATERIAL
T-3727.
Off-white solid.
Sponsor has purity and stability determinations on file.
Store at room temperature.
Any unused test material will be discarded 30 days of issuance of the final report.
Laboratory personnel will take the normal necessary precautions in handling a substance of unknown toxicity. Laboratory clothing, latex gloves, safety glasses, and a particle mask approved for toxic dusts must be worn.
Disposal All waste feed, animal wastes, pan liners, and carcasses resulting from
this study will be disposed of in a high-temperature incinerator (U.S. Smelting Furnace Company, Belleville,Illinois).
2
TEST SYSTEM Animal Model
Young adult male and female albino rats (approximately 7 weeks of age) of the Sprague-Dawley strain will be obtained from Harlan Sprague-Dawley, Madison, Wisconsin. Rats will be selected at random from healthy animals that had been acclimated at HLA for at least 1 week. An adequate number of extras will be purchased in order that no animal in obviously poor health is placed on test. The weight variation in animals used on test will not exceed +20% of the m#an weight (i.e., mean 250 g, range 200 to 300 g).
Reason for Species Selection The rat is the itnimal classically used due to its small size, ready
availability, and large amount of background data.
Identification Each animal will be assigned an individual animal number and ear tag which
will accompany data collected from that animal throughout the study (op-GENB 24).
Housing and Maintenance The following environmental conditions will be maintained in the animal
room used for this study (OP-TARC 230).
Temperature: 22*C +2*
Relative humidity: 50% +20%
Air change: At least 107-changes an hour of filtered 10.0% outsicie air
Light cycle: 12 hours light/12 hours dark
Temperature and humidity will be monitored throughout the study. Variatioas
from prescribed environmental conditions will be documented.
3
Animal husbandry and housing at HLA comply with standards outlined in the ,3 Care will be taken to
"Guide for the Care and Use of Laboratory Animals. ensure that the animals are not disturbed for reasons other than data collection and routine maintenance. The animals will be individually housed, in screerr-bottom stainless steel cages held on racks, with absorbent pan liners in the urine- and feces-collecting pans. Pan liners will be changed at least three times each week.
Feed and water will be provided ad libitum. The diet will be Purina Rat Chow'. No contaminants are expected to be present' in the.feed or water which would interfere and affect the results of the study.
PROCEDURES Experimental Design
Initially, a single dose of 5.0 g/kg will be administered to 10 animals (five males and five females). If no test material-related mortality is produced at this level, no fur'thertesting is required. if any mortality oc curs at the 5.0-g/kg dose level, at the Sponsor's request, three or four geometrically spaced dose levels may be added. Each dose level will consist of 10 animals (five males and five females). Animals will be assigned to groups according to HLA Standard Operating Procedure OP-TOK 42.
Test Material ?reparation and Administration The test material will be suspended in an appropriate vehicle. Individual
dosages will be calculated based upon the animal's body weight taken just before administration of the test material and administered by gavage.
4
Justification of Route of Administration This is the method for administering a known quantity of test substance
and has been the route of choice historically.
Observations The animals wil@ be observed -individually for clinical signs and mortality
at 1.0, 2.5, and 4 hours after test material administration. The animals will be observed daily thereafter for at least 14 days for clinical signs and twice daily (morning and afternoon) for mortality. The 'duration of observations may be extended when considered necessary. The time of death will be recorded as precisely as possible.
Individual body weights will be recorded just prior to study initiation and at 7 and 14 days following test material administration and at death. Changes in body weight will be calculated and recorded when survival exceeds 1 day.
Pathology All test animals, whether dying during the study or sacrificed at
termination, will be subjected to a gross necropsy examination and abnormalities recorded.
Report The final report will contain a description of the test mikterial, a
description of how the study was conducted, response data for clinical signs, mortality and body weights by sex, a discussion of the data, and gross pathology findings.
5
Maintenance of Raw Data and Records Original data or copies thereof will be available at HLA to facilitate
auditing the study during its progress and prior to acceptance of the final report.. When the final report is completed, all original paper data, as well
as the final report, will be retained in the archives of-MA, Madison,
Wisconsin (OP-GEN 44).
REFERENCES
1. Organisation for Economic Cooperation and Development's Guidelines for Testing of Chemicals, Section 401, Acute Oral Toxicity, adopted May 21, 1981.
2. 21 CFR 58. 3. DHEW Publications No,.(NIH) 78-23 (1978).
6
PROTOCOL APPROVAL
Dallas D. Zi;nmerman, FnL) Sponsor's Repreo sentative 3H
Steven M. Glaza Study Director Group Leader, Acute Toxicology Hazleton Laboratories America,
Inc.
(1064S/jg)
Date Date
HAZLEOCON' LABORATORIES AMERICA, INC.
3301 KINSMAN BLVD.
P.O. BOX 7545 MADISON, Wi 53707
(608)241-4471
TLX 7039M HAZRAL MDS UD
PROTOCOL TP2072
Primary Eye Irritation Study in Rabbits (OECD Guidelines) Study No. 50202473
for
3M St. Paul, Minnesota
by'
Hazleton Laboratories America, Life Sciences Division 3301 Kinsman Boulevard
Madison, Wisconsin 53704
Inc.
February 25, 1985 1985, Hazleton IAboratories America, Inc.
Chemical & BioMedical Sciences Division
PROTOCOL TP2072
Primary Eye Irritation Study in Rabbits (OECD Cuidelines)
Study No. Study Location
Test Material Sponsor's Representative Study Director
50202473 Hazleton Laboratories America, Inc. Life Sciences Division 3301 Kinsman Boulevard Madison, Wisconsin 53704
T-3727
Dallas D. Zimmerman, PhD Steven M. Glaza
Proposed Timetable Starting Date Completion Date Final Report Date
Week of February 25, 1985 Week of March 1, 1985 Week of April 1, 1985
OBJECTIVE The objective of,this study is to determine the level of irritation produced following a single exposure of a test material to one eye of albino rabbits. All aspects of this study will conform to the Organ3.sation for Economic Cooperation and Development's Guidelines for Testing Chemicals., Section 405, Acute Eye irritation/Corrosion, Adopted May 12, 1981 and the U.S. Food and Drug Administration's Good Laboratory PractLce Regulations for Nonclinical Laboratory Studies.2 All procedures will be done according to Hazleton Laboratories America, Inc. (MA) Standard Operaring Procedures (SOPS)
referenced in this protocol.
Identification Test -Material: Physical Description: Purity and Stability:
Storage Conditions: Test Material Retention:
Safety Precautions:
TEST MATERIAL
T-3727.
Off-white solid.
Sponsor has purity and stability determinations on file.
Store at room temperature.
Any unused test material will be discarded 30 days after issuance of final report.
Laboratory personnel will take the normal necessary precautions in handling a substance of unknown toxicity. Laboratory clothing, latex gloves, safety glasses, and a particle mask approved for toxic dusts must be worn.
2
TEST SYSTEM Test Animal
Young adult albino rabbits of either sex of the New Zealand White strain, approximately 14 weeks of age, will be obtained from Hazleton Research Products, Inc., Denver, Pennsylvania. An adequate number of extra animals will be purchased so that no animal in obviously poor health is placed on test. The New Zealand White albino rabbit is the animal of choice based upon its large orbit and nonpigmented iris.
Acclimation Upon receipt, the animals will be taken to a designateo animal room where
they will be acclimated for at least 1 week before being placed on test (OP-GENB 36). During acclimation, the animals will be examiued for clinical abnormalities indicative of health problems (e.g., diarrhea, ectoparasites, rough hair coat, nasal or ocular discharge, evidence of injury, etc.). Any animals regarded as unsuitable for the study purposes because of poor physical condition will not be released from acclimation and the reason(s) will be documented.
Identification Each animal in the study will be assigned a permanent identification
number and will be identified with a metal ear tag (OP-GENB 24). All data collected from an animal will be recorded and filed under its identification number.
3
Housing and Maintenance The following environmental conditions will be maintained in the animal
room used for this study (OP-TARC 230).
0 Temperature: 21*C +20
0 Relative humidity: 5OZ +20% 0 Air change: At least 10 changes an hour of filtered 100% outside air
0
Light cycle: 12 hours light/12 hours dark
Temperature and humidity will be monitored throughout the study. Variatioas
from prescribed environmental conditions will be documented.
Animal husbandry and housing at HLA comply with standards outlined in the
"Guide
for
the
Care
and
Use
of
laboratory
.,,3 Animals
Care
will
be
taken
to
ensure that the animals are not disturbed for reasons other than data
collection and routine maintenance. The animals will be housed individually in screen@-bottomstainless steel cages (heavy gauge) held on racks, with absorbent pan liners in the urine- and feces-collecting.pans. Pan liners will
be changid at least three times each week. Feed and water will be provided ad libitum. The diet will be Teklad
Laboratory Rabbit Diet. No contaminants are expected to be present in the feed or water which would interfere and affect the results of the study.
Study Design Three rabbits will be selected at random based upon healttiand a body
weight of 2.0 to 3.5 kg.
PROCEDURES Pre ration and Administration of Test Material
The rabbits' eyes will be examined using fluorescein dye procedures within 24 hours prior to test material administration. Only animals with no sign of
4
corneal injury or eye abnormalitieswill be utilized. one eye of each animal will be treated with the test material and the other eye will serve as the untreated control .
Each rabbit will receive 0.1 g (or the weight equivalent of 0.1 mL) of solid test material. If necessary, the solid test materialswill be finely ground into a dust or powder. The test material will.be placed into the everted lower lid of the rabbit's eye. The upper and lower lids are then to be gently held together for 1 second befo*rereleasing to prevent loss of material. The eyes of the rabbits will remain unflushed for 24 hours following instillation of the test material. After 24 hours, a washout may be used if considered appropriate.
Reason for Route of Administration 4
Historically, the route of choice based on the method of Dra3.ze.
Observations The treated eyes of all animals will be examined for ocular irritation at
1, 24, 48, and 72 hours after treatment. If no irritationor injury is
present at 72 hours, the study will be terminated. ik irritation is present
at 72 hours, additional observations will be made at 96 hours and at 7, 14, and 21 days. If at any of these time points there is no 3.riitation,the study will be terminated. If injury is still present at 21 days, the Sponsor will be-contacted to determine whether the study should continue or be terminated.
After recording the 24-hour observations, sodium fluorescein may be used to
aid in revealing possible corneal injury. irritation will be graded and
4 scored using the Draize ter-hnique(Attachment 1).
All eye abnormalities
will be recorded.
5
All animals that have a damaged eye producing undue stress or discomfort will be sacrificedfor humane reasons after consultingwith the Sponsor.
Body weights will be recorded prior to test material administrationand at weekly intervals throughout the study. Observations and body weights will be recorded in the study notebook.
Pathology All animals,whether dying or sacrificedat study termination,will be
discarded.
Report The finalreport will present a descriptionof the test material,a
descriptionof the test system, dates of study initiationand termination,a summary table showing the irritation data at each observation period, and any special observations that were recorded.
Maintenance of Raw Data and Records Original data or copi.es thereofwill be availableat HLA to.facilitate
auditing the study during its progressand prior to acceptance of the final report. When the finalreport is completed,all original paper data, as well as the final report,will be retained in the archives of HL&, Madison', Wisconsin (OP-GEN 44).
6
REFERENCES 1. "Acute Eye Irritation/Corrosion," OECD Guidelines for Testing Chemicals,
Section 405 (May 12, 1981). 2. 21 CFR 58. 3. DHEW Publications ND. (NIH) 78-23 (1978)'. 4. Draize, J. H., Appraisal_of the Safety of Chemicals in Foods, Druits, and
Cosmetics - Dermal Toxicity, Assoc-Lation of Food and Drug Officials of the U.S., Topeka',Kansas, pp. 49-51 (1959).
7 PROMCOL APPROVAL
Dallas D. Zimmeriban, PhD @ponsor's "pres@ntative 3M
A"*n M. Glaza Study Director Group Leader, Acute Toxicology Hazleton Laboratories America, Inc.
(10 68S/ g)
Date, Date
PROTOCOL - ATTACHMENT 1
(i) Cornea
(A) onneity degree of density(area most dense taken for madiig) 0
No opacity Scatteredor difrUSSarea,detailsor iris clearlyvisible
1
Easily discernible translucent areas, details of iris.
slightly obscured
2
Opalescentarea3vno detailsor iris visible,size of pupil
barely discernible
3
Opaque,iris invisible
(B)
Sg= involved
One quarter(or less),but not zero
Greaterthanone quarter,but lessthanhalf
2
Greater than half, but less than three quarters
3
Greater than three quarters,up to whole,-arta
4
Ax 3 x 5
Total Hwciau2 so
(2)
(A) ILIUU
0
Normal
FOld2 above COrMal, COnggXtICM,swenim, eircumarneal Weation
(anyor all of theseor combinationof any thereof)lrLS still
rewtim to light(slu"Libreactionis positive)
I
So reactionto Light,hemorrhage,gross do2truction(any or all of these) 2
Az 5
Total Maximum 10
(3)
(A) at== (refgr2to PLIpebralconjt=clttze Only)
0
To"*ls acrual V"aels definital7 injected above CCr2LI
I
More diffuse, deeper ae-mson rod, individual TC33013 not
easily discernible
2
Diffuse beefy red
3
go swening
0
Any xwe"4n
above UCMal (includes nictitating membrane)
I
Obvious swelling with partial ever2ion,of lids
2
Swelling with lids about half closed
3
SweLLing idth Li-daabout half closed to completelyclosed
4
N(oC)diDsiscehhaa"rage 0
Any amwmt different from normal (does not include =all amounts
observed in inner canthus of normal animals) --
-
I
Dischargewith saistanirig of the lids and bairs just adjacent
to Lids
-
2
Discharge With 2C13t*niMg Of the lids and hairs, and considerable
ares@ wound the eye
3
Score (A o B * C) x 2
Total Kazimum a 20
The total 2care for the eye is the suc of all scores obtained 'or the cornea, iris, and conj=ctivas.
