Document zo86dmLo7d7D9MOvv58NkGLwR

M D fil Acute Ocular Irritation Test with T-3421 in Albino Rabbits 'i-- \ Experiment No.: 0883EB028 6 Conducted At: Safety Evaluation Laboratory Riker Laboratories, Inc. St. Paul, Minnesota Dates .Conducted: Conducted By: June 21 , 1983 to August 1, 1983 ."T' y L 'L . n\ ' S ' . V w . ' - 1 J- D. M. Markoe, Jr., BS Toxicologis t Study Director y ) i ~>i "J Date Reviewed By: K. D. O'Malley, BS V Senior Toxicologist Acute Toxicology O ~Da te K. L. Ebbens, BS Date Supervisor, Toxicology Testing 000122 1. Summary The results of the acute ocular irritation test conducted from June 21, 1983 to August 1, 1983 at Riker Laboratories, Inc., St. Paul, Minnesota indicate thar T-3421 is moderately irritating (40.0/110.0) to the unwashed eye of the female albino rabbit. A five second and thirty second, contact washed eye procedure ''ere also conducted employing a five liter wash. The irritation ratings for T-3421, using the limited contact procedures, were mildly irritating (15.7/110.0) for the five second and mildly irritating (18.3/110.0) for the thirty second contact procedure. Minimal corneal opacity, iritis and moderate conjunctivitis were produced during the unlimited contact procedure by the one hour evaluation. The irritation subsided to minimal corneal opacity with vascularization (two animals) and slight conjunctivitis (three animals) by the day seven evaluation. T-3421, when allowed a five and thirty second contact, produced iritis and mild conjunctivitis by the one hour evaluation. All irritation subsided by the day two evaluation. Corneal opacity was not observed in any animal of either treatment group. Introduction The objective of this study was to assess the acute ocular irritation properties of T-3421 when instilled into the washed and unwashed 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. 000123 Method and Results a 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 t 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 interval, the cornea, iris and palpebral conjunctiva were examined and graded for irritation and injury according to a standard scoring b system-- . The maximum possible score at any one examination and scoring period 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 iris 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 Tables 4-6. a -- Hazleton Dutchland, Inc., Denver, PA -- Draize: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics (196S). 000124 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 dissipation 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 characteristics and Quality Assurance statement are contained in Appendices I - IV. 000125 Table 1 Eye Irritation Test - Albino Rabbits Treatment Procedure Test Article T-3421 T-3421 T-3421 Number of Animals Evaluated Form Administered 6 waxy solid 3 waxy solid 3 waxy solid Quantity of Test Article Administered 0.1 gm 0.1 gm 0.1 gm Contact Period (seconds) Volume Evaluation Time of Wash Pos t Dose (tap water) Administration unlimited none 1 Hour, 1 , 2 , 3 and 7 Days 5 seconds none 1 Hour, 1, 2, 3 and 7 Days 30 seconds none 1 Hour, 1 , 2 , 3 and 7 Days Table 2 5. Eye Irritation Test - Albino Rabbits Scale of Weighted Scores for Grading the Severity of Ocular Lesions Ocular Tissues__________________________ Description________________________ Conjunctiva 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. Chemosis (B) Any swelling above normal (included nictitating membrane). Obvious swelling with partial eversion of the lids. Swelling with lids about half-closed. Swelling with lids about half-closed tocompletely closed. 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. Draize Grade 1 2 3 1 2 3 4 1 2 3 Score ( A + B + C ) x 2 Total maximum = 20 Cornea 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. 1 2 3 4 1 2 3 4 Score equals A x B x 5 Total maximum = 80 Iris Values (A) Folds above normal, congestion, swelling, circumcorneal 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 destruction (any or all of these). 1 2 Score equals A x 5 Total maximum = 10 Note: The maximum total score is the sum of all scores obtained for the cornea, iris and conjunctiva. 000127 BEST COPY AVAILABLE TABLE 3 EYE IRRITATION TEST - ALBINO RABBITS Classification of Test Materials Based on Eye Irritation Properties 6. Rating Non-Irritating Range 0.0 - 0.5 Practically Non-Irritating Minimally Irritating Mildly Irritating > 0.5 - 2.5 > 2.5 - 15.0 > 1 5 . 0 - 25.0 Moderately Irritating > 25.0 - 50.0 Severely Irritating > 50.0 - 80.0 Extremely Irritating > 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, increase rating one 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 < 2 0 . If 7-day mean score is < 20 but < 60% 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 one 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 < 4 0 . If 7-day mean score is < 40 but < 6 0 % 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. 000128 MB0129 TABLE 4 Examination Tissuu Period Cornua(D-A) 1 Hour Iris Conjunctiva (RSD) Total Cornua(D-A) 1 Day Iris Conjunctiva (RSD) Total Cornea(D-A) 2 Days Iris Conjunctiva (RSD) Total Cornea(D-A) 3 Days Iris Conjunctiva (RSD) Total Cornua(D-A) 7 Days Iris Conjunctiva (RSD) Total 3B988 1 0 (1-2) 5 16(2-3-3) 31 20(1-4) 5 18(3-3-3) 43 20(1-4) 5 12(2-2-2) 37 20(1-4) 5 8 (2- 1- 1) 33 10(1-2)V 0 8 (2- 1- 1) 18 EYE IRRITATION TEST - ALBINO RABBITS with T-3421 3B981 15(1-3) RESULTS ANIMAI, NUMBERS 3B1012 5(1-1) 3B1014 5(1-1) 0 10(2-1-2) 5 14(2-2-3) 14(2-2-3) 25 24 19 20(1-4) 20(1-4) 20(1-4) 5 55 12 (2- 2- 2) 16(3-2-3) 12(2-2-2) 37 41 37 20(1-4) 20(1-4) 20(1-4) 5 55 12(2-2-2) 14(2-2-3) 10(2-1-2) 37 39 35 20(1-4) 5 8 (2- 1- 1) 33 10(1-2)V 0 8 (2- 1- 1) 18 20(1-4) 20(1-4) 55 12(2-2-2) _8J2-1-1)_ 37 33 00 00 2 (1-0-0) _0 20 Key : Cornea : D=Density A=Area Conjunctiva : R=Redness S=Swolling D=Discharge V=Vascularization 3B995 10(1-2) 5 1 0 (2- 1- 2) 25 20(1-4) 5 18(3-3-3) 43 5 12 (2- 2- 2) 37 20(1-4) 5 8 (2- 1- 1) 33_______ 0 0 0 0 3B962 5 (1-1) 0 14(2-2-3) 19 20(1-4) 5 14(2-2-3) 39 1 0 (1- 2 ) 5 12 (2- 2- 2) 27 1 0 (1-2 ) 5 8 (2- 1- 1) 23 0 0 0 Moans 8.3 2.5 13.0 23.8 20.0 5.0 15.0 40.0 18.3 5.0 12.0 35.3 18.3 5.0 8.7 32.0 3.3 0.0 3.0 6.3 000130 TADLE 5 EYF. IRRITATION TEST - ALBINO RABBITS with T-3421 (5 Second Contact) Tissue Cornea (D-A) Examination Period __3B1161 1 Hour 0 RESULTS ANIMAL NUMBERS 3B1164 0 3B1148 0 Iris 55 5 Conjunctiva (MO) Total 12(2-2-2) 17 12(2-2-2) 17 8(2-1-1) 13 Cornea (D-A) Iris Conjunctiva (UD) 1 Day Total 0 5 8 (2-1-1) 13 0 5 6(1-1-1) 11 0 5 6 (2-1-0) 11 Cornea (D-A) 2 Days Iris Conjunctiva (MD) Total Cornea (D-A) 3 Days Iris Conjunctiva (RSD) Total 0 0 0 0 0 0 0 00 00 00 00 0 " " " ..... 00 00 00 Cornea (D-A) 7 Days Iris Conjunctiva (RSD) Total 0 0 0 0 Key: Cornea: D = Density A = Area 0 0 0 0 Conjunctiva: R = Redness S = Swelling D = Discharge 0 0 0 0 HEANS 0.0 5.0 10.7 15.7 0.0 5.0 6.7 11.7 0.0 0.0 0.0 0-0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 000131 TABLE 6 Tiaaue Corn*(D-A) Examination Period 1 Hour Iria Conjunctiva (USD) Total Cornaa(D-A) iria Conjunctiva (MO) 1 Day Total Cornaa(D-A) 2 Days Iria Conjunctiva (MO) Total Cornaa (D-A) Iria Conjunctiva (MO) 3 Days Total Cornaa (D~A) 7 Days Iria Conjunctiva (BSD) Total EYE IRRITATION TEST - ALBINO RABBITS with T-3421 (30 Second Contact) 3B1170 0 RESULTS ANIMAL NUMBERS 3B1162 0 3B1165 0 55 5 12 (2-2-2) 16(3-3-2) 12(2-2-2) 17 21 17 0 5 8 (2-1-1) 13 0 0 0 0 0 5 10(2-2-1) 15 0 0 0 0 0 5 10(2-2-1) 15 0 0 0 0 00 00 00 00 00 00 00 00 0 0 0 0 0 0 0 0 Key: Cornea: D = Density A = Area Conjunctive R = Redness S = Swelling D = Discharge nutr*&Aknicj 0.0 5.0 13.3 18.3 0.0 5.0 9.3 14.3 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 BEST COPY AVAILABLE Riker Experiment No .:_ i^ J Z L j_i __2_______ APPENDIX I PROTOCOL 10. T S T : Acute Ocular Irritation Test__________________________________________________________________________ S P O N S O R : 3 M ____ C o r . : ^ ; i a i C h e m ica l________________________________________________________ D ivisio n C O N D U C TED BY: Safety Evaluation Laboratory, Riker Laboratories, Inc., St. Paul, Minnesota T E S T A R T IC L E :_____ I z l z l l _____________________________________________________________________________ ____ C O N T R O L A R TIC L E : ^ ** ________________________________________________________________________________ P R O P O S E D STA R TIN G /C O M P LETIO N D A TE O F T E S T : . - - t t ? ~ '</ *___________________________________ T E S T S Y S TE M : Female New Zealand White Albino Rabbits SOURCE: O B JE C T IV E : The objective of this test will be to determine the irritation potential of the test article to the ocular tissues (cornea, iris and conjunctiva) of -- i ------------albino rabbits. Rabbits 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 fooc 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 o f ___ : U _____ with the contralateral eye of each animal serving as a control. A t _____ 3____________________ hours and V ' ~ v-.d 7_________days (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 et a/k. After completion of the test, the scores will be analyzed, and a descriptive eye irritation rating assigned to the test article. Eye examinations may be carried out with the aid of sodium fluorescein. If deemed necessary by the study director, washed eye procedures entailing a 5 and 30 second contact period with a I I __ 3 i .Ir. 1t>. wash over a ___ 5 - d n _____ 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. Purina Rabbit Chow, Ralston-Purina, St. Louis, Missouri ^ Draise: Appraisal of the Safety of Chemicals in Foods, Drugs, and C osmetics (1965) Published by the Editorial Committee of The Association of Food an^TDrug Officials of the United States. / p rr {-LA LC e** W. C . .''c C o n n ic k Sponsor w 33 Date Study Director 000132 BEST COPY AVAILABLE Si'or .-oer^en: No. : 0883EB0286 APPENDIX I (concludaci) Deviations and/or Aran cut.ants to Protocol t. The source for the test animals is Hazelton-Dutchland Labs., Denver, PA. 11, - 1- r s. Study Director 6/21/83 Date 2. St udy Director Date 3. Study Director cata 4. Study Director Cate S 000133 Study Director Da te APPENDIX II Principal participating personnel Involved in the Study Name D. M. Markoe, Jr., BS K. L. Ebbens, BS K. D. O'Malley, BS G. C. Pecore Function Toxicologist Study Director Supervisor Toxicology Testing Senior Toxicologist Acute Toxicology Supervisor Animal Laboratory 000134 APPENDIX III 13. BEST COPY AVAILABLE Test and/or Control Article Characterization k T 'S r -/ ^ T c c `/ - Y ) T -2 4 2 -1 1 . The identity strength, uniformity, composition, purity or other per tinent characterizations of the test and/or control substances have been determined and documented as of 3 - f t t <%,/ % v 23 j7 ,y 's / S 2 . The method of synthesis or origin of the test and control substances, including their amount and the method of bioassay (if applicable) is documented. .S yes jr no _____ 3. The stability of the test and/or control substances have been deter mined or will be determined as of fa ~f~u ~i f t. c - 1 The above information and documentation are located in the sponsor's re cords . sponsor - f a s S 3 Date <20 .A Lu c. / i c ^ U i> . Q u Jf - i S <*" 2 , - / y APPENDIX IV QUALITY ASSURANCE STATEMENT Acute Toxicology Laboratory Studies Study No.: 0883EB0 286 This short term study was audited by Compliance Audit and the final report examined against the raw data on August: 29 , 1983 The results of the audit were reported to the study director and to management on August 30, 1983___ . 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 Audit August 30x 1983 Date 000136