Document 7MzZDQzKDMGBd5JJKQkEgoqZe
Acute Ocular Irritation Test with T-2997CoC
in Albino Rabbits
Experiment No.: Conducted At: Dates Conducted: Conducted By:
Reviewed By:
M. T. Case K. L. Ebbens F. D. Griffith W. C. McCormick
0882EB0009
Safety Evaluation Laboratory Riker Laboratories, Inc. St. Paul, Minnesota November 19, 1981 to December 10, 1981
Z) D. M. Markoe, Jr., BS Toxicologist Study Director
j-, - '/`-di'* -- Date
K. L. Ebbens, BS
Date
Supervisor, Acute Toxicology
2_
1.
Summary The results of the acute ocular irritation test conducted from
November 19, 1981 to December 10, 1981 at Riker Laboratories, Inc., St. Paul, Minnesota indicate that T-2997CoC is severely irritating (6 0 .3 / 1 1 0 .0 ) when allowed a five second contact and severely irritating (59.0/110.0) when allowed a thirty second contact.
Mild corneal opacity, iritis and severe conjunctivitis were produced during the five and thirty second contact procedures by the one hour evaluation. The corneal opacity increased to moderate by the one day evaluation.
The irritation persisted and epithelial sloughing and vascularization were noted at the three and seven day evaluations. The fourteen day evaluation revealed diminished irritation and by the twenty-one day evaluation the irritation had generally subsided with the exception of one animal of the five second contact group.
Introduction The objective of this study was to assess the ocular irritation
potential of T-2997CoC to the washed eye of female albino rabbits. This study was conducted in accordance with the Food and Drug Administration s Good Laboratory Practice Regulation of 1978. The raw data generated by the Study Director and the final report are stored in the conducting laboratory's archives.
2.
Method and Results young albino rabbits of the New Zealand breed^ were used to evaluate
the ocular irritating properties of the test article. The test method was modeled after that of Draize et al-.
The test article was instilled into the conjunctival sac of the right eye of each rabbit according to the treatment procedure presented in Table 1 with the left eye of each animal serving as a control. At each scoring in terval, the cornea, iris and palpebral conjunctiva were examined and graded
.b for irritation and injury according to a standard scoring system-. The max imum possible score at any one examination and scoring period is 110 points, which indicates maximal irritation and damage to all three ocular tissues (cornea, iris, conjunctiva) while a score of zero indicates no irritation (Table 2). In this scoring system, special emphasis is placed upon irritation or damage to the cornea, while less emphasis is placed upon damage to the i n s and conjunctiva.
After completion of the test, the scores were analyzed, and a descriptive eye irritation rating was assigned to the test article. The criteria used for assignment of the descriptive rating were the frequency, the extent, and the persistence of irritation or damage which occurred to the three ocular tissues (Table 3). The individual results are presented in Table 4.
a_ Dutchland, Inc., Denver, Pennsylvania b Draize: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics
(1965)
3.
The rating is arrived at by selecting the maximum mean irritation score at one hour, one, two, or three days after instillation. If the rate of dis sipation of injury does not meet the requirements defined for the descriptive rating appropriate for a particular numerical score, the descriptive rating is raised by one or more levels. The rating system is presented in Table 3. The protocol, principal personnel involved in the study, composition char acteristics, and Quality Assurance statement are contained in Appendices I IV.
TABLE 1 EYE IRRITATION TEST - ALBINO RABBITS
Treatment Procedure
Test Article
Number of
Animals
Form
Evaluated_____Administered
T-2997CoC
3
liquid
T-2997COC
3
liquid
Quantity of Test Article Administered______
Contact
Volume of
Period
Wash
(seconds)______ (tap water)
Evaluation Time Post Dose Administration
0.1 ml 0.1 ml
30 5 liters-- 1 Hour, 1, 2, 3, 7, 14 and 21 days.
a 5 5 liters-- 1 Hour, 1, 2, 3, 7,
14 and 21 days.
-- The wash procedure was conducted using a Peri flow Pump, Technilab instruments, Pequannock, New Jersey; at a rate of one liter per minute for five minutes.
Cornea Iris
TABLE 2
EYE IRRITATION TEST - ALBINO RABBITS
Scale of Weighted Scores for Grading the Severity of Ocular Lesions
5.
Opacity (A) Opacity - Degree of density (area which is most dense is taken for reading). Scattered or diffuse area, details of iris
clearly visible. Easily discernible translucent areas, details of iris slightly obscured. Opalescent areas, no details of iris visible, size of pupil barely discernible. Opaque, iris invisible.
Area of Cornea Involved (B) One quarter (or less) but not zero Greater than one-quarter, but less than one-
half. Greater than one-half, but less than three-
quarters . Greater than three-quarters, up to whole area.
Score equals A x B x 5 Total maximum = 80
Values (A) Folds above normal, congestion, swelling, circumcomeal injection (any or all of these or combination of any thereof), iris still reacting to light (sluggish reaction is
positive). No reaction to light, hemorrhage, gross des-
struction (any or all of these).
Score equals A x 5
Total maximum = 10
1 2 3 4
1 2 3 4
Ocular Tissues
Conjunctiva
TABLE 2 (concluded)
EYE IRRITATION TEST - ALBINO RABBITS
Scale of Weighted Scores for Grading the Severity of Ocular Lesions
-- --------------------Description
""
Draize Grade
Redness (A) Redness (refers to palpebral conjunctiva only). Vessels definitely injected above normal. More diffuse, deeper crimson red, individual vessels not easily discernible. Diffuse beefy red.
1 2 3
Chemosis (B) Any swelling above normal (includes nictitating
membrane). Obvious swelling with partial eversion of the lids. Swelling with lids about half-closed. Swelling with lids about half-closed to completely
closed.
1
2 3 4
Discharge (C) Any amount different from normal (does not include small amount observed in inner canthus of normal
animals). Discharge with moistening of the lids and hairs
just adjacent to the lids. Discharge with moistening of the lids and hairs
and considerable area around eye.
1
2 3
Score (A + B + C) x 2
Total maximum = 2 0
6.
Note: The maximum total score is the sum of all scores obtained for the cornea, iris, and conjunctiva.
Rating Non-Irritating
Practically Non-Irritating Minimally Irritating Mildly Irritating
Moderately Irritating
Severely Irritating
Extremely Irritating
TABLE 3
EYE IRRITATION TEST - ALBINO RABBITS
Classification of Test Materials Based on Eye Irritation Properties
7.
Range 0.0 - 0.5 > 0.5 - 2.5 >2.5-15.0 >15.0-25.0 >25.0-50.0
>50.0-80.0
> 80.0 - 110.0
Definition
To maintain this rating, all scores by the one day reading must be zero ; otherwise, increase rating one level.
To maintain this rating, all scores by the one day reading must be zero; otherwise, ir.craaae rating cne level.
To maintain this rating, all scores by the three day reading must be zero; otherwise, increase rating one level.
To maintain this rating, all scores by the 7-day reading must be zero; otherwise, increase rating one level.
To maintain this rating, scores by 7 days must be < 10 for 60% or more of the animals. Also, mean 7-day score must be < 20. If 7-day mean score is < 2 0 but < 6 0 % of animals show scores < 10, then no animal among those showing scores > 10 can exceed a score of 30 if rating is to be maintained; otherwise, raise rating cne level.
To maintain this rating, scores by 7 days must be < 30 for 60% or more of the animals. Also, mean 7-day score must be < 40. If 7-day mean score is < 40 but < 60% of the animals show scores < 30, then no animal among those showing scores > 30 can exceed a score of 60 if rating is to be maintained ; otherwise, raise rating one level.
TABLE 5
Tissue Cornea(D-A)
Examination Period 1 Hour
Iris
Conjunctiva (RSD)
Total
Cornea(D-A)
Iria Conjunctiva
(RSD)
1 Day Total
Cornea(D-A)
Irie Conjunctiva
(RSD)
2 Days Total
Cornea(D-A)
Irie Conjunctiva
(RSD)
3 Days Total
EYE IRRITATION TEST - ALBINO RABBITS with T-2997CoC (Contact time: 5 Seconds)
RESULTS
1B2899 40 (2-4)
ANIMAL NUMBERS 1B2910
20 (1-4)
IP2925 20(1-4)
5 IB (3-3-3) 63
5 18 (3-3-3) 43
5 18(3-3-3) 43
40 (2-4) 5
16 (3-2-3) 61
40(2-4) 5
16 (3-2-3) 61
40 (2-4) 5
14 (3-2-2) 59
40 (2-4) 5
14 (3-2-2) 59
40 (2-4) 5
14(3-2-2) 59
40 (2-4) 5
14(3-2-2) 59
40 (2-4)E 5
12 (3-2-1) 57
40(2-41' 5
14 (3-2-2) 59
40(2-4)E 0
14(3-2-2) 54
MEANS 26.7
5.0 18.0 49.7 40.0
5.0 15.3 60.3 40.0
5.0 14.0 59.0 4o.O
3.3 13.3 56.6
KEY:
Cornea:
D=Density A=Area
Conjunctiva :
R-Redness S-Swelling ^Discharge
E--Epithelial slou^hmo
Tissue Cornea(D-A)
Iris
Examination Period
7 Days
Conjunctiva (RSD)
Total
Cornea(D-A)
Iris Conjunctiva
(RSD)
14 Days Total
Cornea(D-A)
Iris
Conjunctiva (RSD)
21 Days Total
TABLE 5 (concluded)
EYE IRRITATION TEST - ALBINO RABBITS with T-2997CoC (Contact time: 5 Seconds)
1B2893
RESULTS ANIMAL NUMBERS
1B2910
1B2925
20(2-2)E
40 (2-4)E
20(2-2)E
0 00
10 (2-2-1)
12 (3-2-1)
10(2-2-1)
30 52 30
10(2-1)V
30 (2-3)V
10 (2-1)V
0 00
24 (1-0-1)
10(2-2-1)
6(1-1-1)
24 40 16
0
20(2-2)V
5 (l-l)V
0 00
0
8 (1-2-1)
0
0 28 5
MEANS 26.7 0.0 10.7 37.4 16.7 0.0 6.7 23.4 8.3 0.0 2.7 11.0
KEY:
Cornea:
D=Density A=Area
Conjunctiva :
R=Redness S=Swelling D=Discharge
E=Epithelial sloughing V=Vascularization
TABLE 6
Tissue Cornea(D-A)
Iris
Conjunctiva (RSD)
Examination Period 1 Hour
Total
Cornea(D-A)
Iris Conjunctiva
(RSD)
1 Day Total
Cornea(D-A)
Iris
Conjunctiva (RSD)
2 Days Total
Cornea(D-A) Iris
Conjunctiva (RSD)
3 Days Total
EYE IRRITATION TEST - ALBINO RABBITS
with T-2997CoC (Contact time: 30 Seconds)
__________
RESULTS
ANIMAL NUMBERS
1B2898
1B2901
1B2914
40 (2-4)
20(1-4)
20 (1-4)
555
18(3-3-3)
___ 18 (.3-3-3)
18 (3-3-3)
63 40(2-4)
43 40 (2-4)
_4J} 40 (2-4)
555
14 (3-2-2)
14 (3-2-2)
14 (3-2-2)
59 59 59
40(2-4)
40 (2-4)
40 (2-4)
555
14 (3-2-2)
14 (3-2-2)
14(3-2-2)
59 59 59
40 (2-4)E
40 (2-4)E
40 (2-4)E
550
14 (3-2-2)
16(3-2-3)
12 (2-2-2)
59 61 52
MEANS
26.7 5.0
18.0 ______ 49.7________
40.0 5.0
14.0 59.0 40.0
5.0 14.0 59.0 40.0
3.3 14.0 57.3
01
KEY :
Cornea : D=Density A =Area
Conjunctiva : R=Redness S=Swelling D=Discharge
E=Epithelial sloughing
Tissue Cornea(D-A)
Iris
Examination Period
7 Days
Conjunctiva (RSD)
Total
Cornea(D-A) 14 Days Iris
Conjunctiva (RSD)
Total
Cornea(D-A)
Iris Tonjunctlva
(RSD)
21 Days Total
TABLE 6 (concluded)
EYE IRRITATION TEST - ALBINO RABBITS with T-2997CoC (Contact time: 30 Seconds)
RESULTS
____
ANIMAL NUMBERS
1B2898 30(2-3)VE
1B2901 10(2-1)V
__ i m i 4 _________ 2 0 (2-2)V
00
0
10 (2-2-1)
10(2-2-1)
8 (2-1-1)
40 20
28
10(2-1)V
5 (l-l)V
5 (l-l)V
00
0
8 (2-1-1)
4(1-0-1)
6(1-1-1)
18 9
11
5(1-1)
0
0
00
0
00
0
50
0
MEANS 20.0
0.0 9.3 29.3 6.7 0.0 6.0 12.7 1.7 0.0 0.0 1.7
IT
KEY:
Cornea : D=Density A=Area
Conjunctiva : R=Redness S=Swelling D=Discharge
E=Epithelial sloughing V=Vascularization
Riker Experiment No.:
APPENDIX I
PROTOCOL
12.
T E S T. Acute Ocular Irritation Test ;
~ '.-V-- .------------------------------------- ---------------- --------------------------------
-,,
SPONSOR: 3 M--
~' 1 "
r ---------------------------
"'
Division
CONDUCTED BY: Safety Evaluation Laboratory, Riker Laboratories, Inc., St. Paul, Minnesota
TEST ARTICLE: ~ " ' ' -- --------------------- ---------------------------------------------------- '
'
CONTROL A R TICLE:---------------------------------------- ---------------------------------------- '
'
PROPOSED STARTING/COMPLETION DATE OF T E S T :----------------------------------------------------------
TEST SYSTEM: Female New Zealand White Albino Rabbits
SOURCE:
OBJECTIVE:
The obiective of this test will be to determine the irritation potential of the test article to the ocular tissues
[cornea ?r,s and c o n ju n c t) o.
------- a,bino rabbi*. Babbits were selected as the test system
for their sensitivity to irritants, historical use, ease of handling and general availability.
METHOD:
The animals will be housed in standard wire-mesh cages in temperature and humidity controlled rooms with food^ and water offered ad libitum. Each animal will be assigned a numbered ear tag which will correspond to a card affixed to the outside of the cage. The test article will be instilled into
the conjunctival sac of the right eye at a dose of
------- with the contralateral eye of each animal
serving as a control. At _J----- '
--------------- hours and -------- ---------- --------- -- ----- ^
(additional scoring intervals may be added to further characterize the ocular reactions), the tissues will
be examined and graded for irritation and injury according to a standard scoring system of Draize
ef a/fe. After completion of the test, the scores will be analyzed, and a descriptive eye irritation rating assiqned to the test article. Eye examinations may be carried out with the aid of sodium fluoresceim
If deemed necessary by the study director, washed eye procedures entailing a 5 and 30 second
contact period with a - ~ ^ ______wash over a
-------------- period will be conducted using
3 animals per procedure. All raw data generated by the study director and the final report will be
stored in the Riker Laboratories' Archive, St. Paul, Minnesota.
/
k .< y Sponsor
a Purina Rabbit Chow, Ralston-Purina, St. Louis, Missouri b Draise: Appraisal of the Safety of Chemicals in Foods, Drugs, and Cosmetics (1965)
Published by the Editorial Committee of The Association of Food and Drug Officials
of the United States.
A, , , i*... . \y>.
Date
Study Director
v 0,'\V)0 W
Form 19171-7-A.PWO
Hiker Experiment No. : 0P>^> APPENDIX I (concluded) Deviations and/or Amendments to Protocol
7XXfj 13.
aA&ci
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lor^cMXun^.
0~a^ jzQ.
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l/ft
r^Au i& r pL rriizJ cord^r'i ymedu/Si -- U2C^?.i Director
IljujezAAejcQ> (<L-ckr.'uAcAZ&-&-
4U
- K/ iZ l ^ ~ Date
Study Director
Date
Study Director
Date
5.
Study Director
Date
6.
8.
Study Director
Date
APPENDIX II Principal Participating Personnel Involved in the Study
Name D. M. Markoe, Jr., BS
K. L. Ebbens , BS G. C. Pecore
Function
Acute Toxicology Toxicologist Study Director
Supervisor Acute Toxicology
Supervisor Animal Laboratory
APPENDIX III
15.
Test ana/or Control Article Characterization for
_________ T-2997CoG_---------------
1. The identity strenyth, uniformity, composition, purity or other per
tinent characterizations of the test and/or control substances have
been determined and documented as of
7 </
__________
2. The method of. synthesis or origin of the test and control substances, including their amount and the metlxxl of bioasr.ay (if applicable) in
documented.
3 . 'in*. ci-Ahiiitv of the test and/or control substances have been deter
The cords.
om 10703 PWO
RECEIVED
N O V 2 4 1980 jC'n'y Ivskniteu
16.
APPENDIX IV
QUALITY ASSURANCE STATEMENT Acute Toxicology Laboratory Studies
Study No.: .0882EB0009
This short term study was audited by Cou^j.j.ance nuun, ana the final report examined against the raw data on January 25, 1982 . The results of the audit were reported to the study director and to the management on January 25, 1982_____ .
In addition to the data audit, different significant phases for studies underway in the Acute Toxicology Laboratory are inspected weekly on a recurring cycle, and the facilities are examined by Compliance Audit on a three month schedule.
"Compliance Aucfit January 25, 1982
Date
