Leberco*Celsis Testing
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1 n km Leberco*Celss Testng 23 Hawthorne Street Roselle Park, NJ / LABS Fax Nov. 5, 996 SUBMTTED TO: ACTS Testng Labs Buffalo, NY Patty Dck ASSAY NUMBER: DATE RECEVED: /25/96 TEST MATERAL: Renlla Lucderase & Coelenterazne w/table Salt + Sodum Bcarbonate - (composte) Log #: 6B-593 DATE NTATED: /29/96 COMPLETED: H/l/96 PURPOSE OF ASSAY: To determne the degree of eye r r t a t o n, f any, that the test materal may produce when n s t l l e d to the eyes of rabbts, n accordance wth the method descrbed n the Federal Hazardous Substances Act, 6 CFR, Secton RESULTS: Not a prmary eye rrtant. METHOD OF ASSAY: A. ANMALS The albno rabbts used n ths study were obtaned from a USDA lcensed dealer. The anmals were housed ndvdually and acclmated to laboratory condtons for at least four days pror to beng placed on test. Ths toport s submtted for the exclusve use of the person, partnershp or corporaton to whom t s addressed, and nether the report nor the ncme of ft*** Laboratores r«ca arty m*mb*rt of H staff, may us*d f, cof<f#ctof, #ttt, ft* adv6fth<f(j Of t(mt of any product or process wthout wrtten authorzaton. M E M B E R
2 leberco*celss Testng Durng the acclmaton and test perod each anmal was housed and mantaned accordng to The Gude For The Care and Use of Laboratory Anmals (NH 86-23). Each anmal was dentfed by an eartag number and a cage card. The anmals were provded wth Agway Prolab Hgh Fber Rabbt feed and cty water ad lbtum. B. PROCEDURE: Sx normal healthy albno rabbts were used for ths study. On the day of the study pror to testng, each anmal had ther eyes examned for any r r t a t o n or corneal damage; any anmal exhbtng abnormaltes was excluded from testng. Pror to dosng, TThe test materals were composted. Fve-tenths ml of DMSO (Prolume Ltd.) was added to tube of Coelenterazne, then nverted 5 tmes to dssolve. To one tube of Prolume l t d. # whch contaned Renlla L.Ase, NaCl, NaHC3, tap water was added to the 45 ml lne, then nverted tmes to dssolve. To the tube of Coelenterazne & DMSO mxture,.5 ml of Prolume Ltd. "Gn" was added, nverted 5 tmes to mx; then added to the Renlla L.ASE & tap water soluton. Ths resulted n a yellow lqud. Each anmal had. m l l l t e r s of the composted test materal, as suppled, nstlled nto the conjunctval sac of the test eye. The contralateral untreated eye served as a control for that anmal. The treated and untreated eyes were examned and graded at 24, 48 and 72 hours post nstllaton. The crtera for scorng s shown n Table. The ndvdual readngs taken may be found on Table. C. NTERPRETATON OF RESULTS: A test s nterpreted as: Postve - f four or more anmals exhbt a postve response on the cornea, r s or conjunctva. Negatve - f one or less anmals exhbt a postve response on the cornea, r s or conjunctva. nconclusve - f two or three anmals exhbt a postve response on the cornea, r s or conjunctva. The test may be repeated wth a dfferent group of sx anmals and s postve f three or more anmals exhbt a postve response. 2
3 Leberco«Celss Testng SUMMARY: The submtted materal, when tested as a composte n accordance wth the above procedure, produced mnmal conjunctval rrtaton n two of the sx anmals tested. Four anmals had no rrtaton. All sgns of r r t a t o n cleared by day 3 of the study. Therefore, the test materal would not be consdered a prmary eye rrtant as defned by the Federal Hazardous Substance Act, 6 CFR, Secton 5.3 (c) (4). Performed Under the Supervson of:., LEBERCO-CELSS TESTNG Edwn C. Rothsten, Ph.D. Drector or Wllam S. Glman, M.S. Assocate Drector 3
4 Leberco*Celss Testng Sample D #: TABLE Prmary Eye rrtaton Sevres RABBT # Rght Eve. Cornea A. Opacty Total Score = A x B x 5. rs A. Values Total Score - Ax 5. Conjunctvae B. Chemoss C. Dscharge Total Score = (A + B + Total rrtaton Score RABBT n Rght Eve. Cornea A. Opacty Total Score =AxBx5. rs A. Values. Conjunctvae B. Chemoss C. Dscharge Total Score = (A + B + Total rrtaton Score RABBT #3 Rght Eve. Cornea A. Opacty Total Score = Ax B x 5. rs A. Values. Conjunctvae B. Chemoss C. Dscharge Total Score = (A + B + Total rrtaton Score C)x2 C)x2 C)x2 dm Zdav 33m "2! 2 dm 2 day Sdm ff a da? 4m 3dm \6 d \ _ j!!!!!!!
5 Leberco»Celss Testng Sample D #: RABBT Cornea #4 Left Eve A. Opacty. rs A. Values Total Score = Ax B x 5 Total Score - Ax 5. Conjunctvae B. Chemoss C. Dscharge Total Score = (A + B + C)x2 TABLE (Contnued) 4m 2 dm 34m Total Score = (A + B + C)x2 Total rrtaton Score RABBT. Cornea #5 Left Eve A. Opacty L. rs A. Values Total Score = Ax Bx 5. Conjunctvae B. Chemoss C. Dscharge \ $ Total Score = (A + B + C) x 2 Total rrtaton Score RABBT #6 Left Eve. Cornea A. Opacty. rs A. Values Total Score = Ax Bx 5. Conjunctvae B. Chemoss C. Dscharge 2 dm 2dav 3 dm & dm 24m 3 dm l ; j Total Score = (A + B + C) x 2 2 Total rrtaton Score \ \ J J.._.'_ AVERAGE RRTATON SCORES &7" fo!!
6 Leberco«Celss Testng TABLE GRADES FOR OCULAR LESONS CORNEA Opacty: Degree of densty (area most dense taken for readng) No ulceraton or opacty Scattered or dffuse areas of opacty (other than slght dullng of normal luster), detals of rs clearly vsble Easly dscernble translucent area, detals of rs slghtly obscured Nacreous area, no detals or rs vsble, sze of pupl barely dscernble Complete corneal opacty, rs not dscernble RS Normal Markedly deepened folds, congeston, swellng, moderate crcumcorneal njecton (any of these or combnaton of any thereof), rs stll reactng to lght (sluggsh reacton s postve) No reacton to lght, hemorrhage, gross destructon (any or all of these) CONJUNCTVAE Redness: refers to palpebral and bulbar conjunctvae excludng cornea and rs Vessels normal Some vessels defntely njected Dffuse, crmson red, ndvdual vessels not easly dscernble Dffuse beefy red Chemoss: lds and/or ncttatng membranes No swellng Any swellng above normal (ncludes ncttatng membrane) Obvous swellng wth partal everson of lds Swellng wth lds about half closed Swellng wth lds more than half closed Dscharge: No dscharge Any amount dfferent from normal (does not nclude small amounts observed n nner canthus of normal anmals Dscharge wth mostenng of the lds and hars just adjacent to lds Dscharge wth mostenng of the lds and hars, and consderable area around the eye * Starred fgures ndcate lowest grade consdered postve under the Federal Hazardous Substances Act Regulaton 6 CFR, 5.42.
ACTS Testing Labs, Inc. Buffalo, NY Patty Dick. Primary Dermal Irritation in Rabbits. 10/29/96 COMPLETED: H/l/96
Leberco*Celsis Testing 123 Hawthorne Street Roselle Park, NJ 07204-0206 908.245.1933 / 800.523.LABS Fax 908.245.6253 Nov. 5, 1996 SUBMITTED TO: ACTS Testing Labs, Inc. Buffalo, NY Patty Dick ASSAY NUMBER:
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