Document VJXdk8yQ0Z3M3Ov1m7yd4KxDZ

AR226-3213 TRADE SECRET Study Title H-25134: Acute Eye Irritation/Corrosion Study in Rabbits Laboratory Project ID: DuPont-9104 TEST GUIDELINES: OECD Guidelines for Testing of Chemicals Section 4: Health Effects, No. 405 (1987) Commission Directive 92/69/EEC EEC Method B.5 (1992) AUTHOR: Carol Finlay, B.A. STUDY COMPLETED ON: January 3, 2002 PERFORMING LABORATORY: E.I. du Pont de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 Page 1 of 16 Company Sanitized. Does not contain TSCA CBI CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation of the data obtained from this study. yJaimgC.Macfcayn' Reviewed by; ^.}/t^i^ 'TJ^J^^-______________ .Macfcayn Associate Scientist 3'Jif^' ><ftl- - Date Issued by Study Director: j SLA&C ^Ui4itU^i._________ S -^^h -^6 /L Cai6lFinlay,B.A^VDate Staff Scientist 2 Compaq Sanded. Doe3 ^ ^ . TSCA C8J TABLE OF CONTENTS Page CERTIFICATION.........................................................................................................................! STUDY INFORMATION.............................................................................................................4 STUDY PERSONNEL...................................................................................................................5 SUMMARY ....................................................................................................................................6 INTRODUCTION..........................................................................................................................? A. Objective....................................................,...................................,.....................................? B. Principles of the Methodology.............................................................................................7 MATERIALS AND METHODS..................................................................................................7 A. Test Guidelines ....................................................................................................................7 B. Test Substance .....................................................................................................................7 C. Animal Husbandry...............................................................................................................8 D. Protocol................................................................................................................................8 E. Data Analysis and Interpretation of Results.........................................................................9 RESULTS AND DISCUSSION ..................................................................................................11 CONCLUSIONS..........................................................................................................................11 RECORDS AND SAMPLE STORAGE ....................................................................................11 TABLES........................................................................................................................................12 1. DRAIZE SCALE FOR SCORING OCULAR LESIONS...............................................:.................................... 13 2. INDIVIDUAL RABBIT OCULAR IRRITATION SCORES FOLLOWING TREATMENT WITH H-25134 ............................................................................................................................................................... 15 3. MEAN SCORES FOR OCULAR RESPONSES FOR INDIVIDUAL RABBITS.............................................. 16 4. BODY WEIGHTS AND CLINICAL SIGNS OF TOXICITY OBSRVED IN RABBITS.................................. 16 Company Sanitized. Does not contain TSCA CE 3- STUDY INFORMATION Substance Tested: Synonyms/Codes: Submitter's Notebook Numberfs) Haskell Number: 25134 CAS Registry Number: Physical Characteristics: Dark brown solid Sponsor: E.I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A. Study Initiated/Completed: November 28, 2001 / (see report cover page) In-Life Initiated/Completed: December 10,2001 / December 13, 2001 Company San,.,zed. Does no. co.,aln TSCA CB1 4- STUDY PERSONNEL Study Director: Management: Primary Technician: Carol Finlay, B.A. Judith C. Stadler, Ph.D., D.A.B.T. James C. Mackay II Toxicology Report Preparation: Wanda F. Dinbokowitz Laboratory Veterinarian: William Singleton, D.V.M., A.C.L.A.M. Company Sanitized. Does not contain TSCA Cffi 5- SUMMARY H-25134 was evaluated for acute eye irritation potential in 3 adult New Zealand White rabbits. Approximately 100 mg of test substance was administered to 1 eye of each test animal. The eyes remained unwashed following treatment. The conjunctiva, iris, and cornea of each treated eye were evaluated and scored according to a numerical scale approximately 1, 24,48, and 72 hours following administration of the test substance. No irritation was observed in the treated eyes of 2 of the 3 rabbits. Conjunctival redness (score of 1) and discharge (score of 1) were observed one hour after treatment in the treated eye of the remaining rabbit. No irritation was observed 24, 48, or 72 hours after treatment. Fluorescein stain examinations were negative for comeal injury in the treated eyes of all rabbits. No clinical signs were observed, and no body weight loss occurred. Mean values were calculated for each animal separately by using numerical scores obtained from the quantitative evaluation of each ocular response observed in each rabbit approximately 24, 48, and 72 hours following treatment. These values are as follows: RABBIT NUMBER 35417 35408 35414 CORNEAL OPACITY 0.00 0.00 0.00 IRITIS 0.00 0.00 0.00 CONJUNCTIVAL REDNESS 0.00 0.00 0.00 CONJUNCTIVAL CHEMOSIS 0.00 0.00 0.00 Based on the mean values for each rabbit calculated according to the guide for the labeling of dangerous substances published in the Official Journal of the European Communities (EEC Directive 93/21), H-25134 is not classified as irritant. ^^----^cw 6- INTRODUCTION A. Objective The objective of this study was to evaluate the eye irritation potential and the reversibility of ocular effects ofH-25134 following a single ocular application to albino rabbits. The results of the study were used to determine the appropriate toxicity classification and labeling requirements in accordance with EEC (Directive 93/21). B. Principles of the Methodology Ocular effects are evaluated and scored approximately 1,24, 48, and 72 hours after instillation of 0.1 mL of test substance (or a weight of not more than 100 mg of test substance) into the lower conjunctival sac of 1 eye of each of 3 rabbits. The reversibility of any effects is assessed for up to 21 days, if necessary. Ocular effects are scored according to the Draize Scale (Table 1). For each animal, a mean value is calculated for each ocular response by averaging the scores obtained from the 24-, 48-, and 72-hour observations. Classification of ocular imtancy potential is based on the mean values for the lesions observed as well as the persistence of the lesions observed. MATERIALS AND METHODS A. Test Guidelines The study design complies with the following testing guidelines: Organisation for Economic Co-Operation and Development (OECD) (1987). 405 Acute Eye Irritation/Corrosion. Guideline/or Testing of Chemicals. European Economic Communities (EEC) (1992). Directive 92/69/EEC Annex V, Part B.5, Acute Toxicity (Eye Irritation). Methods for the Determination of Toxicity. B. Test Substance The test substance, H-25134, was supplied by the sponsor as a dark brown solid. The test substance appeared to be stable under the conditions of the study. No evidence of instability, such as a change in color or physical state, was observed. S.niferi.^"01""1""150*0" Company 7- H-25134: Acute Eye Irritation/Corrosion Study in Rabbits__ C. Animal Husbandry __________DuPont-9104 Adult HM:(NZW)fBR New Zealand White rabbits were received from Covance Research Products, Denver, Pennsylvania. The rabbits were housed singly in stainless steel, wire-mesh cages suspended above cage boards. Each rabbit was assigned a unique identification number which was recorded on a card affixed to the cage. The last 3 digits of the identification number were written on the inside of each rabbit's ear with a water insoluble marker. The rabbits were offered approximately 125 grams ofPMI Nutrition International, Inc. Certified High Fiber Rabbit LabDiet 5325 daily during the study. Water was available ad libitum. As specified in the Haskell Laboratory animal health and environmental monitoring program, the following procedures are performed periodically to ensure that contaminant levels are below those that would be expected to impact the scientific integrity of the study: Water samples are analyzed for total bacterial counts, and the presence ofcoliforms, lead, and other contaminants. Feed samples are analyzed for total bacterial, spore, and fungal counts. Samples from freshly washed cages and cage racks are analyzed to ensure adequate sanitation by the cagewashers. Certified animal feed is used, guaranteed by the manufacturer to meet specified nutritional requirements and not to exceed stated maximum concentrations of key contaminants, including specified heavy metals, aflatoxin, chlorinated hydrocarbons, and organophosphates. The presence of these contaminants below the maximum concentration stated by the manufacturer would not be expected to impact the integrity of the study. The animal health and environmental monitoring program is administered by the attending laboratory animal veterinarian. Evaluation of these data did not indicate any conditions that affected the validity of the study. Rabbits were quarantined, weighed, and observed for general health for 7 days. Animal rooms were maintained on a timer-controlled, 12-hour light/12-hour dark cycle. Environmental conditions of the rooms were targeted for a temperature of20C 1C and relative humidity of 50% 10%. Excursions outside these ranges were judged to have been of insufficient magnitude and/or 'duration to have adversely affected the validity of the study. D. Protocol On the day of treatment, the eyes of 3 male New Zealand White rabbits were examined using illumination, magnification, and fluorescein stain. Animals used on this study were free of pre existing comeal or conjunctival injury or irritation and were judged to be in good health. The rabbits weighed from 1635 to 2104 grams on the day of treatment. ^s" UO-S"-^0"181"^031 Company Sand. ed,, , ---^- H-25134: Acute Eye Irritation/Corrosion Study in Rabbits___ ____________________DuPont-9104 The container of test substance was heated in a 64C waterbath until the test substance was liquefied. The test substance was then stirred. The weight of a 0.1 mL aliquot of the liquefied test substance was determined to be 119.4 mg. Therefore, approximately 100 mg of test substance was weighed out for each animal. The aliquot of test substance designated for each rabbit was pulverized before dosing. Approximately 100 mg of H-25134 was introduced into the lower conjunctival sac of the right eye of each rabbit. The left eye of each rabbit was not treated with the substance and served as a control. The treated and control eyes of all rabbits remained unwashed. Each rabbit was observed for approximately 30 to 60 seconds before being returned to its cage. Approximately 1,24, 48, and 72 hours after H-25134 was administered, the rabbits were examined for evidence of eye irritation and for clinical signs oftoxicity. At each of these observation periods, eyes were examined using illumination and magnification and scored for ocular reactions according to the Draize Scale (Table 1). Any other adverse ocular reactions, particularly those indicative of corrosive action, were also noted. Control eyes were not scored. These untreated eyes were used for comparison and were considered "normal" relative to the treated eye. Fluorescein stain examinations were conducted at the 24-hour and each subsequent evaluation. The rabbits were weighed on the day of treatment and at the last ocular evaluation. E. Data Analysis and Interpretation of Results The numerical values corresponding to each animal tissue (cornea, iris, conjunctiva) and time were recorded. For each animal, a mean value was calculated for each lesion (comeal opacity, iritis, conjunctival redness, and conjunctival chemosis) by averaging the scores obtained from the 24-, 48-, and 72-hour observations. The results were interpreted according to Annex VI of the EEC Commission Directive 93/21 relative to the general classification and labeling requirements for dangerous substances. Interpretation according to Annex VI - Eye Irritation Criteria: The test substance will be classified as "IRRITANT" and will require the symbol "Xi" and the indication of danger "IRRITANT" in accordance with the criteria below. 'In addition, the following risk (R) phrases will be assigned to substances classified "IRRITANTS" as applicable, according to the following criteria: R36 "IRRITATING TO EYES" The risk phrase, R36 "IRRITATING TO EYES", will be assigned if, when applied to the eye of the test animal, the test substance causes significant ocular lesions which occur within 72 hours after exposure and which persist for at least 24 hours. Ocular lesions are significant if the individual animal mean values of the 24-, 48-, and 72-hour evaluations in 2 or more animals comply with any of the following criteria: __ ^-"---"'"" _^_ H-25134: Acute Eye Irritation/Corrosion Study in Rabbits________________________DuPont-9104 - comeal opacity equal to or greater than 2.0, but less than 3.0 - iridial lesion equal to or greater than 1.0, but less than 2.0 - conjunctival redness equal to or greater than 2.5 - conjunctival chemosis equal to or greater than 2.0 R41 "RISK OF SERIOUS DAMAGE TO THE EYES" The risk phrase, R41 "RISK OF SERIOUS DAMAGE TO THE EYES", will be assigned if the test substance causes severe ocular lesions which occur within 72 hours after exposure and which persist for at least 24 hours. Ocular lesions are severe if the individual animal mean value of the 24-, 48-, and 72-hour evaluations in 2 or more animals comply with any of the following criteria: - comeal opacity equal to or greater than 3.0 - iridial lesion equal to 2.0 R41 "RISK OF SERIOUS DAMAGE TO THE EYES" risk phrase is also required if: - ocular lesions are still present 21 days following treatment - the test substance causes irreversible coloration of the eyes % noiconia'-"^081 Sa"^0085"010 Company 10- RESULTS AND DISCUSSION No irritation was observed in the treated eyes of 2 of the 3 rabbits. Conjunctival redness (score of 1) and discharge (score of 1) were observed one hour after treatment in the treated eye of the remaining rabbit. No irritation was observed 24,48, or 72 hours after treatment. Fluorescein stain examinations were negative for comeal injury in the treated eyes of all rabbits. No clinical signs were observed, and no body weight loss occurred. The ocular scores from individual animals with respect to observation time are presented in Table 2. Mean ocular scores calculated in accordance with EEC Directive 93/21 are summarized in Table 3. Individual body weights and clinical signs oftoxicity are presented in Table 4. CONCLUSIONS Based on the mean values for each rabbit calculated according to the guide for the labeling of dangerous substances published in the Official Journal of the European Communities (EEC Directive 93/21), H-25134 is not classified as irritant. RECORDS AND SAMPLE STORAGE Specimens (if applicable), raw data, and the final report will be retained at Haskell Laboratory, Newark, Delaware, or at Iron Mountain Records Management, Wilmington, Delaware. Company S. ani.t.iz.e^d- n"oow-s, n" ot contain TSCACBI 11 - TABLES Company Sanitized. Does not contain TSCACB1 - 12- TABLE 1 DRAIZE3 SCALE FOR SCORING OCULAR LESIONS (1) Cornea (A) Opacity-degree of density (area most dense taken for reading) No opacity....................................................................................... 0 Scattered or diffuse area, details of iris clearly visible.................... 1 Easily discernible translucent areas, details of iris slightly 2 obscured.................................................................................... Opalescent areas, no details of iris visible, size of pupil barely 3 discernible............................................................................... Opaque, iris invisible....................................................................... 4 (B) Area of cornea involved One quarter (or less) but not zero.................................................... 1 Greater than one quarter, but less than half..................................... 2 Greater than one half, but less than three quarters.......................... 3 Greater than three quarters, up to whole area.................................. 4 (2) Iris (A) Values 0 Normal............................................................................................. 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)............ 1 No reaction to light, hemorrhage, gross destruction (any or all of 2 these)......................................................................................... (3) Conjunctiva (A) Redness (refers to palpebral and bulbar conjunctiva excluding cornea and iris) Vessels normal................................................................................ 0 Vessels definitely injected above normal...............'......................... 1 More diffuse, deeper crimson red, individual vessels not easily 2 discernible................................................................................. Diffuse beefy red...................................;......................................... 3 Company Sanitized. Does not contain TSCA CQ/ 13- TABLE 1 (Continued) DRAIZE SCALE FOR SCORING OCULAR LESIONS (B) Chemosis No swelling..................................................................................... 0 Any swelling above normal (includes nictitating membrane)......... 1 Obvious swelling with partial eversion of lids................................ 2 Swelling with lids about half closed ............................................... 3 Swelling with lids about half closed to completely closed ............. 4 (C) Discharge No discharge.................................................................................... 0 Any amount different from normal (does not include small amounts observed in inner canthus of normal animals)............ 1 Discharge with moistening of the lids and hairs just adjacent to 2 lids............................................................................................. Discharge with moistening of the lids and hairs, and considerable area around the eye............................................... 3 a Draize, J. H., "Dermal Toxicity." Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics. The Editorial Committee of the Association of Food and Drug Officials of the United States, Austin, Texas, 1959, pp. 46-59. I Company Sanitized. Does not contain TSCA CB/ 14- 1-25134: Acute Eye Irritation/Corrosion Study in Rabbits TABLE 2 INDIVIDUAL RABBIT OCULAR IRRITATION SCORES FOLLOWING TREATMENT WITH READING 1 HOUR 24 HOURS 48 HOURS 72 HOURS RABBIT NUMBER 35417 35408 35414 35417 35408 35414 35417 35408 35414 35417 35408 35414 CORNEA OPACITY AREA 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 IRITIS 0 0 0 0 0 0 0 0 0 0 0 0 REDNESS 1 0 0 0 0 0 0 0 0 0 0 0 CONJUNCTIVA CHEMOSIS DI 0 0 0 0 0 0 0 0 0 0 0 0 -15- Coinpa"^ S.nifc. TABLES MEAN SCORES" FOR OCULAR RESPONSES FOR INDIVIDUAL RABBITS RABBIT NUMBER 35417 35408 35414 CORNEAL OPACITY 0.00 0.00 0.00 IRITIS 0.00 0.00 0.00 CONJUNCTIVAL REDNESS 0.00 0.00 0.00 CONJUNCTIVAL CHEMOSIS 0.00 0.00 0.00 a 24hr+48hr+72hr/3. Calculated according to EEC Directive 93/21. TABLE 4 INDIVIDUAL BODY WEIGHTS OF RABBITS RABBIT NUMBER 35417 35408 35414 SEX Male Male Male INITIAL WEIGHT (g) 2028 1635 2104 FINAL WEIGHT (72 HOURS) (g) 2192 1752 2208 CLINICAL SIGNS OF TOXICITY OBSERVED IN RABBITS No clinical signs oftoxicity were observed in any of the rabbits during the study. Company Sanitized. Does ->l contain TSCA CB) 16-