Document 4aNozbmXdoz1azgJk0Q3yNrZa

foe-HAZLETON LAECRATOF4ES AMEFIC-A. 3301 KINSMAN BLVD., P.O.BOX 7545 MADISON, Wl S3707 USA FINAL REPORT Karen Armstrong 3M Toxicology Services Building 220-2E-02 St. Paul, MN 55144-1000 Sample: T-4416 HLA Number: 91201739 CA APPRP, Enclosed: Acute Oral ToxicityStudy in Rats (OECD Guidelines) o Key Personnel o Methodology o Summary of Results o Individual Pathology Comments o References o Pathology Report o Raw Data/Appendix Signed: Steven M. Study Director Acute Toxicology PHONE (608)241-4471 FACSIMILE (608)241-7227 TELEX TLX 703956 HAZRAL L4DS UD Date a OORNING Lxbwatory Services Con"ny 1):HAZLETON LABC)RATORIES AN4EFNCA. r4c. 3301 KINSMAN BLVD., P.0, BOX 7545 MADISON, Wi 53707 USA HLA Number: 91201739 Sample: T-4416 Acute Toxicology Steven M. Glaza Study Director Section Supervisor Steven R. Sorenson Study Coordinator Patricia Padgham Group Leader Sharen L. Howery Report Coordinator Ouality Assurance Deborah L. Keller Manager Page 2 KEY PERSONNEL Laboratory Animal Veterinarian Cindy J. Cary, DVM Diplomate, ACLAM Anatomical Patholoqy Thomas E. Palmer, PhD Anatomical Pathologist Dennis Hoffman Group Leader Necropsy Anne Mosher Group Leader Pathology Data PHONE (608)241-4471 FACSIMILE (608)241-7227 TELEX TLX 703956 HAZRAL MDS UD 8C)ORPOYD Labmawy Serwcm Cwnpay HAZLETON LABC)RATCRIESANO'ERC-A@r-'C. 3301 KINSMAN BLVD., P.O.BOX 7545 MADISON, Wi W707 USA HLA Number: 91201739 Sample: T-4416 Page 3 METHODOLOGY Objective: To assess the acute oral toxicity produced when a test material is administered by oral gavage to rats.' Test Material: T-4416 Physical Description: Purity and Stability: Storage and Retention: Safety Precautions: Clear colorless liquid Sponsor assumes responsibility for purity and stability determinations (including under test conditions). The test material was stored at room temperature. Any unused material will be returned to the Sponsor according to HLA Standard Operating Procedure. Normal handling procedures were used according to HLA Standard Operating Procedure. Test Animal: Adult albino rats, Crl:CDOBR, were procured, separated by sex, maintained in group cages in temperature- and humidity-controlled quarters, provided continuous access to Rodent ChowO 5001, Purina Mills, Inc., and water, and held for an acclimation period of at least 7 days. Animal husbandry and housing at HLA comply with standards outlined in the "Guide for the Care and Use of Laboratory Animals".2 If variations from the prescribed environmental conditions existed, they were documented and considered to have no effect on the study outcome. No contaminants were expected to have been present in the feed or water which would have interfered with or affected the results of the study. Acclimated animals were chosen at random for the study. Five male and five female rats, weighing from 204 to 299 g, were used for each dose level. The study consisted of three dose levels (0.2, 1.0 and 5.0 g/kg of body weight). Test animals were housed by sex in groups of five and identified by animal number and corresponding ear tag. Food and water were available ad libitum throughout the study, except for an overnight period just before test material administration when food, but not water, was withheld. Reason for Species Selection: Historically, rats have been used as a representative of a rodent species and are preferred by various regulatory agencies. Preparation and Administration of Test Material: The test material was administered as received. An individual dose of the undiluted material was calculated for each animal based upon its fasted body weight and administered by gavage. The dose volume of the test material varied for each dose level based upon the bulk density of the material of 1.28 g/mL. PHONE (608)241-4471 FACSIMILE (608)241-7227 TELEX TLX 703956 HAZRAL MDS UD aOC)F4f4WJLOabwalarSywvcm COffMny HAZLETON LABC)FTATORF-S AMEFICA. 3301 KINSMAN BLVD., P.O. BOX MADISON, Wi S3707 USA f'JC. 7545 HLA Number: 91201739 Sample: T-4416 Page 4 Reason for Route of Administration: Historically, the oral route has been the route of choice for administering a known amount of test material. Observations: The animals were observed for clinical signs and mortality at 1, 2.5 and 4 hours after test material administration. The animals were observed daily thereafter for 14 days for clinical signs and twice daily for mortality. Animals were weighed just before test material administration. Surviving animals were weighed again at Day 7 and at termination of the experimental phase or at death (when survival exceeded I day). Pathology: All animals whether dying on test or euthanatized at termination of the experimental phase were subjected to a gross necropsy examination and all abnormalities were recorded. After necropsy, the animals were discarded and no tissues were saved. Statistical Methods: The LD value for males, females and the combined sexes was calculated based upon t%e observed mortality using a computer program utilizing a modified Behrens-Reed-Muench Cumulant Method .3 No other statistical analysis was performed. Location of Raw Data and Final Report: The raw data and a copy of the final report will be retained in the archives of HLA. PHONE (608)241-4471 FACSIMILE (608)241-7227 TELEX TLX 703956 HAZRAL MDS UD 0 CORNW40 LabMSWV S@" CDRWnny MAZLOTON LABORATOPAES APAEF%C'&- t'4r-' 3301 KINSMAN BLVD., P.O.BOX 7545 MADISON, Wi 53707 USA HLA Number: 91201739 Sample: T-4416 Page 5 SUMMARY OF RESULTS Test Animal: Albino Rats - Crl:CDOBR Source: Charles River Laboratories, Inc., Portage MI Date Animals Received: 12/04/89, 12/18/89 and 01/08/90 Method of Administration: Oral Gavage Start Date (In-life): 12/28/89 End Date (In-life): 02/16/90 Estimated Oral LD,, : Male - 1.8 g/kg of Body Weight 95% Confidence Limits - 0.7 to 4.9 g/kg Female - 0.6 g/kg of Body Weight 95% Confidence Limits - 0.2 to 1.5 g/kg Sexes Combined - 1.0 g/kg of Body Weight 95% Confidence Limits - 0.4 to 2.4 g/kg Sex Male Female Dose Level (q/kq) 0.2 1.0 5.o 0.2 1.0 5.0 - Average Body Weights (g) Initial Dav 7 Terminal 290 332 362 272 264 315 274 --- --- 211 247 254 233 257 268 230 --- --- Mortality (Number Dead/Number Dosedl 0/5 1/5 5/5 0/5 4/5 5/5 PHONE (6os)241-4471 FACSIMILE (WS) 241-7227 TELEX TLX 703956 HAZRAL MDS UD C>DRr4w4Q LabW*$Wy Sdwvom C@mr"ny ePluevH"A3Z@LlCMCN LABOF:tATORF-SAN-iEF4CA-r@f-' 3301 KINSMAN BLVD., P.O. BOX 7545 MADISON, Wi S3707 USA HLA Number: 91201739 Sample: T-4416 Page 6 Observation Appeared normal Clinical Signs (Number of Animals Affected) Hour 1.0 2.5 Males (0.2 q/kq) Day .2 10 Ll 12 13 14 5 5 5 555555555 5 5 5 5 5 Appeared normal Females (0.2 q/kq) 5 5 5 55555 5 5 5 5 5 5 5 5 5 Males (1.0 q/kq) Appeared normal Hypoactivity 5 5 5 52 1 1 1 2333 3 3 3 4 4 0 0 0 03 2 222 1 1 1 0 0 0 0 0 Red-stained face 0 0 0 0 1 2 2 2 1 1 1 1 1 1 0 0 0 Yellow-stained urogenital area 0 0 0 0 1 3 3 3 1 1 1 1 1 1 1 0 0 Ataxia 0 0 0 00 00 0 1 1 1 1 0 0 0 0 0 Thin appearance 0 0 0 000001 1 1 1 1 0 0 o o Diarrhea 0 0 0 00 000 00 1 1 0 0 0 0 0 Death 0 0 0 0 1000 o0 oo o o o o o Females (1.0 q/kq) Appeared normal Hypoactivity 5 5 5 511111111 1 1 1 1 1 0 0 0 02 2 100000 0 0 o o 0 Red-stained face 0 0 0 0 2 2 1 0 0 0 0 0 0 0 o 0 0 Ataxia 0 0 0 001 000000 0 0 0 0 o Prostration Death 0 0 0 0 00 1 000 00 0 0 o o o 0 0 0 0 12 1 00000 0 o 0 0 0 PHONE (W8) 241-4471 FACSIMILE (608)241-7227 TELEX TLX 703956 HAZRAL MDS UD HAZLETON LABC)RATDRF-S ANAERCAr4-3301 KINSMAN BLVD., P.O.BOX 7545 MADISON. Wi S3707 USA HLA Number: 91201739 Sample: T-4416 Page 7 Clinical Signs (Continued) (Number of Animals Affected) Observation Hour Day 1.0 2.5 i.0 I Z a 1 5 @k Z a 2 10 11 l@2 l@3 14 Males (5.0 q/kq) Appeared normal 1 0 0 0 -------- - - - - - Diarrhea 4 4 4 1 -------- - - - - - Hypoactivity 0 4 4 1 -------- - - - - - Red-stained face 0 2 4 1 - - - - - - - - - - - - - Lacrimation 0 0 1 0 -------- - - - - - Ataxia 0 0 0 1 -------- - - - - - Death 0 0 1 4 -------- - - - - - One animal found dead after the morning observation. Females (5.0 q/kq) Appeared normal Diarrhea Hypoactivity 2 0 0 0 -------- - - - - 3 3 2 0 -------- - - - - 1 3 2 0 -------- - - - - - Lacrimation 0 2 2 0 -------- - - - - - Ataxia 0 1 1 0 -------- - - - - - Red-stained face 0 1 1 0 -------- - - - - - Death 0 2 1 2 -------- - - - - - PHONE (608) 241-4471 FACSIMILE (608) 241-7227 TELEX TLX 703956 HAZRAL MDS UD a C)ORNWJQ LaboratorSyer@en Cowoony HAZLETON LAEOP"TORES AK4ET4C-ok r4r-' i3OiKINSMAN 13LVD.P,.O.BOX 7545 MADISON, Wi 53707 USA HLA Number: 91201739 Sample: T-4416 INDIVIDUAL PATHOLOGY COMMENTS Dose Level: 0.2 g/kg of Body Weight Animal Test Day Number Sex Died Sacrificed Necropsy Observation C50366 m - 14 No visible lesions C50367 m - 14 No visible lesions C50363 m - 14 No visible lesions C50369 m - 14 No visible lesions C50370 m - 14 No visible lesions C50501 F - 14 No visible lesions C50502 F - 14 No visible lesions C50503 F - 14 No visible lesions C50504 F - 14 No visible lesions C50500 F - 14 No visible lesions Page 8 PHONE (6W) 241-4471 FACSIMILE (608)241-7227 TELEX TLX 703956 HAZRAL MDS UD m OORNW@G Lobwasory S@ Com"ny HAZLETON LABOFLATORF-S AH@ERCA- 3301 KINSMAN BLV[).,P.O.BOX MADISON. Wi s37o7 usA t4C-' 7545 HLA Number: 91201739 Sample: T-4416 Page 9 INDIVIDUAL PATHOLOGY COMMENTS (continued) Dose Level: 1.0 g/kg of Body Weight Animal Test Day Number Sex Died 'Sacrificed- Necropsy Observation C49861 m 14 No visible lesions C49862 m C49863 m - C49864 m 2 C49860 m - C49921 F 3 14 No visible lesions 14 No visible lesions - Stomach, Glandular Mucosa: diffusely red 14 No visible lesions - Stomach, Glandular Mucosa: diffusely red C49917 F 2 No visible lesions C49918 F 4 C49919 F 3 C49920 F - Perinasal Discharge: red Stomach, Glandular Mucosa: multiple dark areas, up to 2 x 4 mm - Stomach, Glandular Mucosa: diffusely red 14 No visible lesions PHONE (608)241-"71 FACSIMILE (608)241-7227 TELEX TLX 703956 HAZRAL MDS UD A OORNWOO LabwolorSy@ ConV*ny HAZLEOCON LABC)RATORF-S AHAERC.A. r4C. 3301KINSMAN BLV[).P,.O.BOX 7545 MADISON. Wi 53707 USA HLA Number: 91201739 Sample: T-4416 Page 10 INDIVIDUAL PATHOLOGY COMMENTS (continued) Dose Level: 5.0 g/kg of Body Weight Animal Test Day Number Sex Died Sacrificed Necropsy Observation C49191 m I - No visible lesions C49192 m I - No visible lesions C49193 m I - Stomach, Glandular Mucosa; multiple brown areas, up to 3 x I mm C49194 m 0 - Stomach, Glandular Mucosa: diffusely dark red C49490 m I - No visible lesions C49131 F 0 - Stomach contents: clear white fluid C49132 F 1 - Stomach, Glandular Mucosa: multiple brown areas, up to 4 x 2 mm C49128 F 0 - Stomach Contents: clear white fluid C49129 F 0 - Stomach Contents: clear white fluid C49130 F 1 - No visible lesions PHONE (608)241-4471 FACSIMILE(608)241-7227 TELEX TLX 703956HAZRAL MDS UD a CX@ORNW.L0aboralary SWAM Company HAZLETON @@W) LABOF2ATC)FIESAA,*F:KC-A-r-,r3301 KINSMAN BLVD., P.O. BOX 7545 MADISON. Wi S3707 USA HLA Number: 91201739 Sample: T-4416 Page 11 REFERENCES 1. "Acute Oral Toxicity," Organisation for Economic Cooperation and Deve7opment's Guide7ines for Testing of Chemicals, Section 401 (adopted May 12, 1981). 2. NIH Publication No. 86-23 (revised 1985). 3. Thakur, A. K. and Fezio, W. L., "A Computer Program for Estimating LDSO and its Confidence Limits Using a Modified Behrens-Reed-Muench Cumulant Method," Drug and Chemical Toxicology, 1(3):297-305 (1981). PHONE (608)241-4471 FACSIMILE (608)241-7227 TELEX TLX 703956 HAZRAL MDS UD 4 C)ORNW40 LOOMaWy SwvCoa CaMnny HAZLETON LABCY:;ATCW%ES AMERC.A. tJC. 3301KINSMAN BLVD.,P.O.BOX 7545 MADISON, Wl S3707 USA HLA Number: 91201739 Sample: T-4416 Page 12 PATHOLOGY REPORT There were 10 rats (5 males, 5 females) from each of 3 dose levels (0.2, 1.0, and 5.0 g/kg) necropsied. The animalsthat died on test (DOT) were refrigeratedand necropsiedwithin 24 hours, usuallythe same day. All survivinganimalswere euthanatizedand necropsiedat the terminationof the study. The dose level, day of death, and gross observationsrecorded for each animal are on page 8 through10 of this report. All animalsin the 5.0 g/kg dose group died on test. At necropsy,the only changes observedwere in the animalsthat died on test. The most frequently recordedobservationspertainedto the content and colorationchanges in the mucosa of the stomach. These findingspossiblyrepresenttest material mixed with ingesta,postmortemchange,or both. All remainingobservationsin the DOTs were considered postmortem change, incidental,and unrelated to the test material. There were no visiblelesionsin the sacrificedanimals. Thomas E. Palmer, PhDPathologist (91201739.cjr) -3 ?C,; Date PHONE (608)241-4471 FACSIMILE (608) 241-7227 TELEX TLX 703956HAZRAL MDS UD 9 (X)RNWJG LabotalarSyervc" cornpany HLA: q/z,-i-7311@ PERSOt4liEL SIGTIATURE SliEET ACUTE TOXICOLOGY Name Becky Beckwith Job Title Lab Animal Technician Signature Initials Ann Bursaw Sr. Clerk Anthony Cass Lab Animal Assistant Cindy J. Cary, DVM Lab Animal Veterinarian I- U@kd@t@ Kari Gassen Sr. Lab Animal Assistant it@n J?I Steven M. Glaza Section Supervisor Ben Haley Sr. Lab Animal Assistant Jeff Hicks Lab Animal Technician cl,C- A LZ J ]Lt) .77-7/ Sharen L. Howery Research Assistant Jim Jirschele Lab Animal Assistant Wayne Madison Senior Section Supervisor Bud McDonald Sr. Lab Animal Assistant Cqwl Eileen McConnell Administrative Clerk 7 Henry Mikula Lab Animal Assistant Albert Oleson Patricia Padgham Sr. Lab Animal Assistant Group Leader /I.., Jane Polnow Lab Animal Technician Steven R. Sorenson Study Coordinator 41 zSahli ,TI. - Dawn Thieding @lerk Annette R. Turner Sr. Clerk 14"tL lei t@-) (09/11/89) cmn In 0 rj ci rJ CD rci Li LA C,2 is 92 lu CL C,2 ex C3 o 0 Gn ma C7. tz cz ci ra ol -1@ @ CP w .0 a 11 CL. w 0 'A4 C2 ol 2, -,2 0 0 -@ cr 0 olo+l ci di cn cn lzr@ E cu E 00 a) cl 0 >E u V) G) w fe to cm 0) ra a u 2= > lm > L- 7;;-o Li a) n mcw Ln la 0.- > c 0 %A 0 0) to 4, = u .:; . 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