Document vy2O9mQL3dg69JMEepb5Dm2rY

2 6 - *3*23 c: C 0 UD REPORT ACUTE EYE IRRITATION/CORROSION STUDY WITH T-7809 IN THE RABBIT ro <-- C3 D=1 UD C"--oD3 -impr0On -- om Bo O CD 03 NOTOX Project 365333 NOTOX Substance 123723 - Page 1 of 10 - i T-7809 NOTOX Project 365333 STATEMENT OF GLP COMPLIANCE______________________________ ____ _________ __ NOTOX B.V., 's-Hertogenbosch, The Netherlands The study described in this report has been correctly reported and was conducted in compliance with the most recent edition of: The OECD Principles of Good Laboratory Practice which are essentially in conformity with: United States Environmental Protection Agency (FIFRA). Title 40 Code of Federal Regulations Part 160. United States Environmental Protection Agency (TSCA). Title 40 Code of Federal Regulations Part 792. United States Food and Drug Administration. Title 21 Code of Federal Regulations Part 58. Japanese Ministry of Agriculture, Forestry and Fisheries. 59 NohSan, Notifications No. 3850. Japanese Ministry of Economy, Trade and Industry. Kanpogyo No. 39 Environmental Agency, Kikyoku No. 85. Japanese Ministry of Health, Labor and Welfare. Ordinance No.21. Study Director: Drs. M.S. Teunissen Management: W.J.A.M. Frieling DVM Date: C & 2Oo3> T-7809 NOTOX Project 365333 QUALITY ASSURANCE STATEMENT_______________________________ _____ ________ NOTOX B.V., 's-Hertogenbosch, The Netherlands This report was audited by the NOTOX Quality Assurance Unit to ensure that the methods and results accurately reflect the raw data. The dates of Quality Assurance inspections and audits are given below. During the on-site inspections procedures applicable to this type of study were inspected. DATES OF QAU INSPECTIONS/ AUDITS on-site inspection(s) 1 1 - 1 4 November 2002 (Process) protocol inspection(s) 23 December 2002 (Study) report audit(s) 15 April 2003 (Study) REPORTING DATES 14 November 2002 (SPF Animal Unit) 23 December 2002 15 April 2003 Head of Quality Assurance: C.J. Mitchell B.Sc. Date: T-7809 NOTOX Project 365333 SUMMARY Acute eye irritation/corrosion study with T-7809 in the rabbit. The study was carried out based on the guidelines described in: EC Commission Directive 92/69/EEC, B.5, "Acute Toxicity - Eye irritation", OECD No.405, "Acute Eye Irritation/Corrosion" and US EPA, OPPTS 870.2400, Acute Eye Irritation, EPA 712-C-98-195, August 1998. Single samples of approximately 51 mg of T-7809 (a volume of approximately 0.1 ml) were instilled into one eye of each of three rabbits. Observations were made 1, 24, 48 and 72 hours after instillation. Instillation of the test substance resulted in effects on the cornea, iris and conjunctivae. Corneal injury was seen as opacity (maximum grade 2) and epithelial damage (maximum 65% of the corneal area) during the observation period. Iridial irritation (grade 1) was observed during the observation period. Irritation of the conjunctivae was seen as redness, chemosis and discharge during the observation period. Necrosis of the nictating membrane was observed in one animal on days 3 and 4 and in two animals on days 2, 3 and 4. In addition, necrosis of the eyelids was seen in one animal on days 3 and 4. Since sufficient data were available, the study was terminated after 4 days for ethical reasons Based on the presence of necrosis of the eyelids and/or nictating membranes, it was concluded that ocular corrosion had occurred by instillation of T-7809 into the rabbit eye in all three animals. Based on these results and according to the EC criteria for classification and labelling requirements for dangerous substances and preparations (Council Directive 67/548/EEC), T7809 should be labelled as: risk of serious damage to eyes (R 41 ). (O T-7809 NOTOX Project 365333 PREFACE Sponsor Study Monitor Testing Facility Study Director Study Plan 3M Belgium N.V. Haven 1005 Canadastraat 11 B-2070 ZWIJNDRECHT Belgium Dr. A Seacat 3 M Medical department 3M Center, Building 220-2E-02 I-94 and Me NOTOX B.V. Hambakenwetering 7 5231 DD 's-Hertogenbosch The Netherlands Drs. M.S. Teunissen Start: 20 January 2003 End : 23 January 2003 TEST SUBSTANCE The sponsor is responsible for all test substance data unless determined by NOTOX. Identification CAS Number Description Batch Purity Test substance storage Stability under storage conditions Expiry date Specific Gravity T-7809 332350-93-3 White crystals #1 See Certificate of Analysis At room temperature in the dark Stable 01 June 2003 1.23 TEST SUBSTANCE PREPARATION The test substance was ground to a powder using a mortar and pestle prior to weighing PURPOSE AND RATIONALE The purpose of this acute eye irritation/corrosion study was to assess the possible irritation or corrosion potential when a single dose of the test substance was placed in the conjunctival sac of the rabbit eye. This study should provide a rational basis for risk assessment in man. The absence of eye pigmentation in the albino rabbit facilitates the evaluation of induced eye reactions. The ocular route was selected because the test substance may accidentally come into contact with the eyes during manufacture, handling and/or use. 1 T-7809 NOTOX Project 365333 GUIDELINES As required by the Dutch Act on Animal Experimentation, the study protocol was reviewed and agreed by the Article 14-functlonary and the Ethical Committee of NOTOX (DEC NOTOX 97-0310) as required by the Dutch Act on Animal Experimentation (February 1997). The study procedures described in this report were based on the following guidelines: European Community (EC), Council Directive 67/548/EEC, Annex V, Part B, Methods for the Determination of Toxicity, as last amended by Commission Directive 92/69/EEC, B.5: "Acute Toxicity - Eye Irritation". Official Journal of the European Communities No. L 383, 1992 Organisation for Economic Co-operation and Development (OECD), OECD Guidelines for Testing of Chemicals, Section 4, Health Effects, No.405, "Acute Eye Irritation / Corrosion", Paris Cedex, 1987. United States Environmental Protection Agency (EPA). Health Effects Test Guidelines, OPPTS 870.2400, Acute Eye Irritation. Office of Prevention, Pesticides and Toxic Substances (7101), EPA 712-C-98-195, August 1998. ARCHIVING NOTOX B.V. will archive for at least 10 years raw data, protocol, report and test substance reference sample. No data will be withdrawn without the sponsor's written consent. TEST SYSTEM Species Number of animals Age and body weight Identification Albino Rabbit, New Zealand White, (SPF-Quality) Recognised by international guidelines as the recommended test system (e.g. EC, OECD) Source: Charles River Deutschland, Kisslegg, Germany 3 Animals of one sex. Animals used within the study were at least 6 weeks old and body weights were at least 1.0 kg. Earmark. ANIMAL HUSBANDRY Conditions A controlled environment was maintained in the room with optimal conditions considered as being approximately 15 air changes per hour, a temperature of 213C, a relative humidity of 30-70% and 12 hours artificial fluorescent light and 12 hours dark per day. Deviations from the maximum level for relative humidity (with a maximum of 20%) occurred which might have been caused by cleaning procedures in the room. Based on laboratory historical data these deviations were considered not to have affected the study integrity. Accommodation Individually in labelled cages with perforated floors (Scanbur, Denmark, dimensions 56x44x37.5 cm). Acclimatisation period was at least 5 days before start of treatment under laboratory conditions. 8 T-7809 NOTOX Project 365333 Diet Standard laboratory rabbit diet (Charles River Breeding and Maintenance Diet for Rabbits, Altromin, Lage, Germany) approx. 100 g. per day. Certificates of analysis were examined and retained in the NOTOX archives. In addition, pressed hay (BMI, Helmond, the Netherlands) was provided twice a week. Water Free access to tap-water. Certificates of quarterly analysis were examined and retained in the NOTOX archives. TREATMENT A health inspection was performed prior to commencement of treatment, to ensure that the animals were in a good state of health. Special attention was paid to the eyes, which were free from any abnormality. Based on the results of the skin irritation study (no irritation noted), the study was started with three animals. Each animal was treated by instillation of 51.3 mg (50.5 - 52.2 mg) of the test substance (a volume of approximately 0.1 ml) in the conjunctival sac of one of the eyes after gently pulling the lower lid away from the eyeball. The lids were then gently held together for about one second to prevent loss of the test substance. The other eye remained untreated and served as the reference control. Immediately after the 24-hour observation, a solution of 2% fluorescein in water (adjusted to pH 7.0) was instilled into both eyes of each animal to quantitatively determine corneal epithelial damage. When considered necessary, this procedure was repeated to assess recovery. Any bright green stained area, indicating epithelial damage, was estimated as a percentage of the total corneal area. OBSERVATIONS MortalityA/iability Toxicity Body Weight Necropsy Irritation Twice daily. At least once daily. Day of treatment (prior to instillation) and at termination. No necropsy was performed on animals sacrificed for severe irritation/corrosion of the eye. The eyes of each animal were examined approximately 1, 24, 48 and 72 hours after instillation of the test substance. The irritation scores and a description of all other (local) effects were recorded. The irritation was assessed according to the following numerical scoring system. At each observation, the highest scores given were recorded: CORNEAL IRRITATION Opacity: degree of density (area most dense taken for reading) No ulceration or opacity (may include slight dulling of normal lu s tre ).....................................................0 Scattered or diffuse areas of opacity, details of iris clearly visible........................................................... 1 Easily discernible translucent area, details of iris slightly obscured........................................................ 2 Nacreous area, no details of iris visible, size of pupil barely discernible................................................3 Opaque cornea, iris not discernible through the o pacity............................................................................ 4 T-7809 NOTOX Project 365333 Area of cornea involved: No ulceration or opacity.......................................................................................................................................0 O ne quarter or less but not zero ....................................................................................................................... 1 G reater than one quarter, but less than half................................................................................................... 2 G reater than half, but less than three quarters.............................................................................................. 3 G reater than three quarters, up to whole a r e a ..............................................................................................4 IRIS Normal ..................................................................................................................................................................... 0 M arkedly deepened rugae, congestion, swelling, moderate circumcomeal hyperaemia, or injection, any of these or combination thereof. Iris still reacting to light (sluggish reaction is positive)..............................................................................................................................1 No reaction to light hemorrhage, gross destruction (any or all of these)...............................................2 CONJUNC TIVAL IRRITA TION Redness (refers to palpebrae and sclera, excluding cornea and Iris): Blood vessels normal .......................................................................................................................................... 0 Som e blood vessels definitely hyperaemlc (injected).................................................................................. 1 Diffuse, crimson color, individual vessels not easily discernible.............................................................. 2 Diffuse beefy re d .................................................................................................................................................... 3 Chemosis (refers to lids and/or nictitating membranes): No sw elling.............................................................................................................................................................. 0 Any swelling above normal (Includes nictitating m em branes)...................................................................1 Obvious swelling with partial eversion of lid s................................................................................................. 2 Swelling with lids about half closed .................................................................................................................. 3 Swelling with lids more than half closed...........................................................................................................4 Discharge: No discharge (may include small amounts observed in inner canthus of normal a n im a ls )............. 0 Any amount different from normal and/or lacrimation...................................................................................1 Discharge with moistening of the lids and hairs just adjacent to lids....................................................... 2 Discharge with moistening of the lids and hairs (considerable area around the e y e ) ....................... 3 W here standard lighting was considered inadequate for observing minor effects, eye examinations were performed using an ophthalmic examination lamp. In cases of equivocal results when comparing the treated and untreated eyes, the illustrated guide from the Consumer Product Safety Commission, Washington, D.C. 20207 was used for additional control purposes. INTERPRETATION The results were evaluated according to the EC criteria for classification and labeling of dangerous substances and preparations (Council Directive 67/548/EEC and all adaptations to technical progress and amendments of this Directive published in the Official Journal of the European Communities). PROTOCOL DEVIATIONS Deviations from the minimum level of temperature occurred (with a maximum of 1C). Based on historical laboratory data this deviation was considered not to have affected the study integrity. T-7809 NOTOX Project 365333 RESULTS Irritation and corrosion (Table 1) Instillation of approximately 51 mg of the test substance (a volume of approximately 0.1 ml) Into one eye of each of three rabbits resulted In effects on the cornea, iris and conjunctlvae. Corneal injury was seen as opacity (maximum grade 2) and epithelial damage (maximum 65% of the corneal area) during the observation period. lridial irritation (grade 1) was observed during the observation period. Irritation of the conjunctivae was seen as redness, chemosis and discharge during the observation period. Necrosis of the nictating membrane was observed in one animal on days 3 and 4 and in two animals on days 2, 3 and 4. In addition, necrosis of the eyelids was seen in one animal on days 3 and 4. Since sufficient data were available, the study was terminated after 4 days for ethical reasons Colouration / Remnants No staining of (peri) ocular tissues by the test substance was observed. Toxicity / Mortality No symptoms of systemic toxicity were observed in the animals during the test period and no mortality occurred. CONCLUSION Based on the presence of necrosis of the eyelids and/or nictating membranes, it was concluded that ocular corrosion had occurred by instillation of T-7809 into the rabbit eye in all three animals. Based on these results and according to the EC criteria for classification and labelling requirements for dangerous substances and preparations (Council Directive 67/548/EEC), T7809 should be labelled as: risk of serious damage to eyes (R 41). u T-7809 NOTOX Project 365333 TABLE 1: INDIVIDUAL EYE IRRITATION SCORES Time after dosing S No 89# Comea Opacity Area Fluor area (%) Iris Conjunctivae Redness Chemosis Discharge Comments 1 hour 24 hours 48 hours 72 hours 12 1 2 4 2- 2 3 65 1 3 4 2 - 23 1 3 4 2B 2 3 55 1 3 4 2 B S No 90# 1 hour 24 hours 48 hours 72 hours 12 1 34 2 3 65 1 3 4 23 1 34 2 3 55 1 3 4 2. 2B 2 AB 2 AB S No 91# 1 hour 24 hours 48 hours 72 hours 12 1 2 4 2- 2 2 50 1 34 2B 21 1 3 4 2B 2 1 25 1 3 3 2 BF Fluor area (%): green staining (percentage of total comeal area) after fluorescein treatment. Comments: A Grey/white discolouration of the eyelids (sign of necrosis) B Grey/white discolouration of the nictitating membrane (sign of necrosis) F Reduced elasticity of the eyelids. TABLE 2: MEAN VALUE EYE IRRITATION SCORES Animal # Comeal opacity Mean 24 - 72 hours Iris Conjunctivae Redness Chemosis 89 2 1 3 4.0 90 2 1 3 4.0 91 2 1 3 3.7 # Animal specifications: Animal no Sex 89 <? 90 91 <? Age at start (weeks) 11-13 11-13 11-13 Body weights (grams) prior to application at termination 2537 2582 2348 2341 2376 2470 u