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-4 HAZLOTON L"BDPffDRIES ANFERICA INC Chemica&l BioMedicaSlcienceDsivision @'N@iol orK.I-NSsMoAN BLVD. - P.O.BOX 7545 * MADISON, WISCONSIN 63707 PHONE (6082)41-4471* TLX 703956HAZRAL MDS UD FINAL REPORT ROGER PERKINS MINNESOTA MINING & TOXICOLOGY ST. PAUL, SERVICES MN 55101 MANUFACTURING COMPANY PURCHASE ORDER NUMBER: T837389-410 712 SAMPLE NUMBER: 70100355 SAMPLE ENTERED: REPORT PRINTED: 01/07/'8-/-@ 01,1,.)/-/ ENCLOSED: PRIMARY EYE IRRITATION STUDY IN RABBITS RAW DATA APPENDIX METHOD,, summAR@, SIGNED: I..................... STEVEN M. GLAZA STUC,@' DIRECTOR ACUTE TOXICOLOG@' ........ DATE ......... BY AND FOR HAZLETON LABORATORIES AMERICA, INC. RAW DATA AMERICA, FOR THIS STUDY INC., MADISON, ARE KEPT ON FILE WISCONSIN. AT HAZLETON LABORATORIES r@@.-'HAZLETONLMBDR=URir=S AKAERICA, INC Chemical& BloMedicalSciencesDivision MADISON, WISCONSIN 53707 - PHONE (6082)41-4471o TLX 703966HAZRAL MDS UD 3301 KINSMAN BLVD. - P.O.BOX 7645 SAMPLE NUMBER: 70100355 SAMPLE: T-4016 PAGE EYE IRRITATION Objective: To determine the level of irritation single exposure of a test material to one eye according to the U.S. Environmental Protection Testino Pesticides and Toxic Substances. produced of albino Agencyl5 following rabbits Guidelines a for Test Material: T-4016 Physical De5cription: Purity and Stability: Thick brown liquid Spon5or a55ume5 responsibility stability determinations. for purity and Test Animal: Young adult rabbits of the New Zealand procured, maintained individually in 5creen-bottom and humidity-controlled quarters, provided access and a measured amount of Purina High Fiber Rabbit an acclimation period of at least 7 days. White strain were cages in temperatureto water ad libitum Chow, and held for Six acclimated animals, weighing from 2354 to 2514 g, were ch05en at random for the test. The animal5' eyes were examined within 24 hours prior to test material administration Using sodium fluorescein dye procedures. Onlv those animals with no sign of ocular injury or irritation were used on the test. Test animals were identified by animal number and corresponding ear tag. Reason for Species animal of choice Selection: based upon The New Zealand White albino rabbit 15 the its large orbit and nonplqmented -'Iris. Preparation of Test Material; was not determined. The sample was dosed as received. The pH Treatment: Each rabbit received 0.1 ml of the liquid test material placed on the everted lower lid of one eyey with the contralateral eye serving as the untreated control. The upper and lower lids were gently held together for one second to prevent 1055 Of material and then released. The eyes of the rabbits remained unflU5hed. Reason for based on Route of Administration: the method of Draize. Historically, the route of choice HAZLETON 3301 KINSMAN BLVD. - P.O. BOX 7545 - MADISON, WISCONSIN Chemical& BioMedicalSciencesDivision 53707 9 PHONE (608) 241-4471 o TLX 703956 HAZRAL MDS UD PAGE SAMPLE NUM13ER: 70100355 SAMPLE: T-4016 EYE IRRITATION (CONTINUED) Ob5ervation5: The treated eyes were ob5erved for ocular irritation at 11 24, 48, 72 and 96 hours after treatment. At the 72-hour reading, POS5ible corneal injury. the Draize technique. sodium fluorescein Was Irritation Was qraded used and to aid scored in revealing accordinq to Animals were weighed iU5t prior to test material administration. Pathology: discarded. At study termination all animals were euthanatized and HAZLOOCON LMX3=.XrC)RIES APVERICA. lrlf- Chemical& BioMedicalSciencesDivision ,*i-- - 3301KINSMAN BLVD. - P.O.BOX 7545 - MADISON, WISCONSIN 53707- PHONE (6082)41."71 - TLX 703956HAZRAL MOS UD QAMPLE NUMBER: 70100355 SAMPLE: 7-4016 PAGE 4 EYE IRRITATION SUMMARY (CONTINUED) Test Animal: Albino rabbits - New Zealand White Source: Hazleton Research Products, Inc., Denver PA Date Animals Received: 12/16/86 Date Test Started: 01/08/67 Date Test Compieted: Oi,-12,,87 PRIMAR@' EYE IRRITATION OBSERUATION PERIOD 1 Ho u r : 24 Hours: 48 Hours: 72 Hours: 96 Hours: SCORES* 6 Rabbit Mean 0.1 ml (Unwashed) 22.7 25.7 12.5 5.7 0.0 The Primary for all the observation Eye Irritation Score is the total eye irritation animals divided by the number of animals (6) at period. score each "-4AZLE'tCNL-M)R@IES "IC 3301 KINSMAN BLVD. - P.O. BOX 7545 - MADISON, WISCONSIN Chemical& B!oMedicalSciencesDivision 53707 o PHONE (608)241-4471 o TLX 703956 HAZRAL MDS UD SAMPLE NUMBER: 701003155 SAMPLE: T-4016 EYE IRRITATION Animal Number FI'0208 F16219 F16220 F16221 F16222 F16223 Observation Period I Hour 24 Hours 48 Hours 72 Hours 96 Hours 1 Hour 24 Hours 48 Hours 72 Hours 96 Hours 1 Hour 24 Hours 48 Hours 72 Hours 96 Hours I Hour 24 Hours 48 Hours "-@2 Hours 96 Hours I Hour 24 Hours 48 Hours 72 Hours 96 Hours I Hour 24 Hours 48 Hours 72 Hours 96 Hours PAGE 5 (CONTINUED) Table I Individual Eye Irritation Scares Cornea A B 0 0 1 3 1 1 0 0 0 0 Score AXE3x5 0 15 5 0 0 Iris Score A A x 15 1 5 1 13 0 0 0 0 0 0 Conjunctivae A B c 3 4 3 2 3 2 2 0 2 1 0 0 0 0 0 0 0 1 5 3 4 1 20 1 5 3 4 3 1 1 5 1 5 3 3 1 0 0 0 0 0 2 2 1 0 0 0 0 0 0 0 0 0 0 0 1 5 3 3 1 1 15 1 5 3 0 0 0 1 5 2 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 5 3 4 1 1 5 1 F, 3 4 1 0 0 0 1 5 2 1 1 0 0 0 0 0 2 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 5 3 14 0 0 3 2 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 i 3 -:> 0 0 0 1 0 0 0 0 0 2 0 1 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 Score (A+B+C)'@, 211 14 6 0 18 20 14 10 0 16 12 e 6 0 18 16 e 6 0 le 14 8 4 0 16 8 4 2 0 HAZLETON L-M)-@IES AfV*F;ICA. ltJC Chemical& B!oMedicalSciencesDivision TLX 703956HAZRAL MDS UD 3301 KINSMAN BLVD. - P.O.BOX 7545 - MADISON, WISCONSIN 53707o PHONE (6082)41-4471 SAMPLE NUMBER: 70100355 SAMPLE: T-4016 EYE IRRITATION (CONTINUED) Sodium Table -? Fluorescein Examination Animal Number Fib2O8 F16219 F16220 F16221 F16222 F16223 Observation Pre-initiation NEG NEG NEG NEG NEG NEG Period 7el Hours 114r,- L@ NEG NEG NEG NEG NEG NEG = No stain retention POS = Positive 5tain retention (area of cornea involved). PAGE 6 Comments: No pain response (vocalization) W85 instillation of the test material. elicited from any animal follot@iinq Blanching of the conjunctivae was seen in all six animals at 1 and 24 hours, in three animals at 48 hours anc in two animals at /7,1h1o-urz,. Corneal epithelial peelinq was exhibited by four animals at 24 hours and by two animals at 48 hours. References: 1. Hitr-Fi,R*.K..,"Primary Eye Irri*atior, Study," Pesticide Assessment Guidelines, Subdivision F, Hazard Evaluation: Human and Domestic AniM8151, U.S. Environmental Protection Hqency Office of Pesticide and Toxic Substances Series 81-4, pp. 51-55 (November 1982). Draize, J.H. , "Apprai5al of the Safety of Chemicals in Food5, DrUq5, and Cosmetics - Dermal Toxicitv." A550Ciation of Food and Drug Officials of the U.S., pp. 46-@9 (19/-5). 3. 21 CFR 58. 4. DHHS Publication No. (NIH 85-23 19e'.',)Guide for the Care and U5e of Laboratory Animals. 01 (A) 22a= - d.ip 80 of dw0itY takm for reading) (aros MOST dms* 0 go opacity early vi"blo 1* S"tterea or d:LffUSOU'*&, dgt&il* Of irS C' Easily dincerniblg trunSIUCOnt-&rOsb,.detail*Of iri& 2* Ali rLtIrODSOLU opal*mmt &rwo, ED det&Uo of iris viable, siSO Of'PUPLL 3* barely discernible Opaquw4.iris invis:LblAw (31 ACISACMZM 'r-'ve* one quarter (or lase), but not =rO Grmtar th@ am quarter,but IOM halr 2 3 Gr"ter "- balf, but 1022 th" throw Quartw2 Orvoter tbm t%L@" Tiartem, uck tv vb0l*'-Ars& Ax Bz Tatal Maxl@ (Zi I:La abom DWML. omwap"ao@. MOLUM. airamowo-LL wastim (mv or ALL of theme or amoblmtlm ae mr tbw4wf) irl& XLLIL - I fo reactin to Light (31,.gg:krmatim to PO@dt:Lve)-- . me rematim to ll&M, banarruaw, sp damtroutus (MW or a" 2* of thme) ASS (3) ZMIM"UM (A) I&== (rorwe to palpobml omi=wt iv@ =IT) ToneeLsnorm" TosaWA dnfi"telr Injected abom owml Mm dlfram. doopm win=& easily dlmoernible r". iRWIT16=1 TG=DOLD got Difrom booty red (B)Mona" nosweLLlag Any vj*LUng above no-ml Cloolud4o act:Ltating saWA-MO) 2 Obviam sualling idth partial wwwsim of IL" 3 sweLLtm w:LtiL Lids abo" hiLLr alas" 3waLlin with Li" about b&U alcg" to eampletaLT alowd (C) 2LWJLN= So discharge Any amomt difrer=t commi (dow not include =MLLL ammmts obmwvod I* inner canthus or norml ani=Lls) MUaftrge wU& maditealm of the 11" and ;ai@ j@L adjamt. 2 to Li" MUmbarge w:Lth m"stenin of the Li" and bairs, a" oomddw2blw are& -ratod the We 3 a 4. C) z I Totakl morinew 7b* total war* for the @" Is the @ aormm, U-U. and eWmetAvedi@. of &U sear@ to St&cred fixuree indicato.positive 4ffoct. obtained for the L( c c 0 -cr tn cr at cm C-i = cm 0. LLi LLS = -CC 3c LLJ cn as 0) C12 u !@ a) P-4 LLJ -i x ui L@ r;; I 0 d) 4-) 4J ea So S- Ect cu = 0- to In r- 0 talc S0 4- u S- $4) a) = 4-3 (U 0 im 0 4-) a) (a d) CU r_ rL= cu S- to a) 4J = ai 4-J S- 0 ai = 3: 4-) r- 0 tv s- 4-) 0 u CU 4) >1 4.; tm S- tu to (1)cu 4J tn = r- 0 Ol.- cu u 0) ID C; a Ul) S- lu cu 4a 3: 0 In 0 to a *0 = 4n u S- ic Q) uw r_ 10 x LA.1 CL (L) E ai u tn d) (L) (L) - 4-J S- .0 cu CK CU 4) ,rL to- cm 4A 4..)ea C, (L)0 0 == = (U 0 ;'-= cu to "49' CU cn cc ui C) V) 0 = 0 cu 0) k 4z1 tm 4-1 4- >S- 0 (U u ai (U t C4 - LLJ (U is to 9= vy 4-) cn -cc 311 P-0 a a r= UA 00 00 e--Oo, Ln -c0u1 to op LL- cu '3 0 31 'A 0 72 - 4-4 cc 4-A IV rlo 3C 00 0 0 to tu Primary Eye Irritation Test Observations est Material: Test Eye: Right HLA No. Group: Washed: PA@ seconds following instillation of test material, the test eye was washed with mL of lukewarm tap water for NO, seconds. L,@nwashed Ariim&T No. Location Of rnrneal Lesions (9 ol C) Tail ---- Head Cornea - OpacitY Area Iris Conjunctivae- Redness Chemosis Discharge Sodium Fluorescein Examl tJA- OBSERVATIOlt PERIOCI'a 19 houv- Fr I A16 AIP t> N)4 ILIA TechnicianDate 2tt!l NA Not Applicable A Petite hemorrhage B alanching C Ciear disr-harge 0 - Purulent discharge E - Hair loss around the eye F - Necrotic Areas G - Unable to *.,4,sL:a7%'41,1ut-co-,- severe swelling H - No reaction to light I - Injected j KL1# 14 PR- SPOS NEG - Corneal epithelial damage, Cornea.1 epithelial damage, Corneal epithelial damage, liypopyon Corneal neavascularizatioit Pannus Unable to visualize due to ::cve-e opa=it.,, Granulation scar tissue Positi-ve-stainretention Negative stain retention peeling piling pitting C)eviewed By. (4253A) Date.- Eye I"itation Score:lz SFAA.- Primary Eye Irritation Test Observations est Material: Test Eye: Right LAC)tLO HLA No. Group: Washed: xj# seconds following instillation of test material, the test eye was washed with mL of lukewarm tap water for L& seconds. L.4n@w a s h e d OBSERVATION PERIOD: Animal No. to (0 Location of Tc ail Head Cornea - Opacity Are& Iris ,@njCuonnjcutnctivae - Redness Chemosis Discharge Sodium Fluorescein Exa-I Technician- Date I-@l-'97 rilvi I @l's C, t.P,,) N Pr Npr-.- N C> N4 I NA - Not AppTicable A - Petite hemorrhage B - Blanching C - Clear discharge 0 - Purulent discharge E - Hair loss around the eye F - Necrotic Areas ','.,abtleo vis,-.al".Zdc:;--c; severe swelling H - No reaction to light - Injected jKLRItPR- SPas NEG - Corneal epithelial damage, Corneal epithelial damage, Corneal epithelial damage, Hypo-pyofv Corneal neavascularization Paffnus Unable to visualize due to savere opacity Granulation scar tissue Positi-ve-stainretention Negative stairt retention peeling piling pitting viewed By: (4253A) Date: -am% Eye Irritation Score: Primary Eye Irritation Test Observations 10 est material: Test Eye: Right HLA No. Group: ---K)pr Washed: ALP, seconds following instillation of test material, the test eye was washed with mL of lukewarm tap water for &I seconds. L,-iTnwa sh ed Animal No. Location or Corneal Lesions OBSERVATION PERIOD'- qg hour.5 'FL--(901CI)OsR19 (aI;t0li (.all Tail ---- Head C) Cornea - Opacity Area Iris C> ,@riCjounnjcuntcit.ivvaaee - Redness Chemosis Discharge C) Sodium Fluorescein Exam m,4 N)q Technician- A) A- A3 A)A Date 0 C) AJI'@ NA - Not Applicable A - Petite hemorrhage R - glanching C - Clear discharge D - Purulent discharge E - Hair loss around the eye F - Necrotic Areas G - Unable to -.IsL:alize-4L:cta severe swetling H - No reaction to light I - Injected J Corneal epithelial damage, peeling K Corneal epithelial damage, piling L - Carceal epithelial damage, pitting 14 - Hypopyon - N - Corneal neovascularizatiorr P - Pannus R - Unable to visualize due to severe op-"--@lity S - Granulation scar tissue POS - Positi-ve-stain retention NEG - Negative stain retention eviewed By.(4253A) Date: Eye Irritation Score: 7i4@ Primary Eye Irritation Test Observations est Material: Test Eye: Riqht HLA No. Group: Qpf N,4 Washed: _&J_4seconds following instillation of test material, the test eye was-washed with mL of lukewarm tap water for && seconds. L@4n@washed OBSERVATION PERIOD: Animal No. Location of Corneal Lesions Ft- to a 19 - (Ollaao Tail Cornea Head Opacity Area n 10 Iris Conjunctivae - Redness 9L Chemosis Discharge Sodium Fluorescein Examl C- N)ea Technician 17> Date gLgLi I ;I 3 tpf,N@, NA - Not Applicable A - Petite hemorrhage B - Blanching C - Clear discharge D - Purulent discharge E - Hair loss around the eye F - Necrotic Areas r. - Una-hieto visualize due to severe swelling H - No reactionto light I - Injected J - Corneal epithelial damage, peeling K - Corneal epithelial damage, piling L - CorneaT epithelial damage, pitting 14 - Hypdpyon N - Corne&T neovascularizatiort P - Pannus R - Unable to visualize due to saverp opacity S - Granulation scar tissue POS - Positive-stainretention NEG - Negative stain retention iewed By-. o rv (4253A) Date: Eye Irritation Score-4#moil-z%-%at Primary Eye Irritation Test Observations est material: Test Eye: Riqht HLA No. Group: opt NPC Washed: ALA. seconds following instillation of test material, the test eye was washed with ML of lukewarm tap water for &L seconds. L@@nwashed OBSERVA7rO N P ER JO D.- n u r-s Animal Na. Location or Corneal Lesions Ft- Tail ---- Head (OCIOR toGiict (0;@.10 t. 9Lal Cornea - Opacity Area Iris conjunctivae - Affft Redness Chemosis Discharge S dium FluoresceinExam A).A AZA *'f-Nl AJ4 Aliq-.AJff Al lechnician- tr;-Oy@ Date NA - Not Applicable A - Petite hemorrhage 8 - Blanching C - CTear discharge D - Purulent disch ge E - Hair loss around the eye F - Necrotic Areas G - Un!ti)let.,vnisualize die,*tn severe swelling H - No reactioffto light I Injected ,_Oev iewed By.(4253A) Date: J - Corneal epithelial damage, peeling K - Corneal epithelial damage, piling L - Corneal epithelial damage, pitting 14 - Hyp6pyon 4 Corneal neovascularization P Pannus R Unable to visualize due to se-,!rtopxctty S Granulation scar tissue POS Positi-vestain retention NEr3- Negativestain retention Eye Irritation Score: I-zt-fill.