REPORT 

SANITIZED 

ACUTE ORAL TOXICITY WITH 

1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE 

IN THE RAT (FIXED DOSE METHOD) 

/"--

NOTOX Project( . NOTOX Substanc

~/ 

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This report contains the unpublished results of research sponsored by Akzo Nobel Chemicals B.V .. Reproduction, issue or disclosure to third parties in any form is not permitted without prior written authorization from the sponsor. 

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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: 

The United States Food and Drug Administration. Title 21 Code of Federal Regulations Part 58. 

The United States Environmental Protection Agency (FIFRA). Title 40 Code of Federal Regulations Part 160. 

The United States Environmental Protection Agency (TSCA). Title 40 Code of Federal Regulations Part 792. 

study Director: Dr.Ir. W.R. Pels Rijcken 

~--~ 

------

Management: Drs. W.J.A.M. Frieling Section Head General and Industrial Toxicology 

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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/ REPORTING DATES 

AUDITS 

on-site inspection(s) 

19 December 1996 

protocol inspection(s) 

2 December 1996 

report audit(s) 

29 January 1997 

19 December 1996 

2 December 1996 

29 January 1997 

Quality Assurance Manager: C.J. Mitchell B.Sc. 

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The study was carried out in accordance with OECD Guideline No. 420, "Acute Oral Toxicity - Fixed dose method" and EEC Directive 92/69/EEC, Part 8.1-bis, "Acute Toxicity-Oral, Fixed dose method". 

Initially, 1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE was administered by oral gavage to one female at 2000 mg/kg body weight. As no mortality occurred and no signs of toxicity were observed, the main study was conducted with a fixed dose of 2000 mg/kg body weight administered to five rats of each sex. The animals were subjected to daily observations. Body weights were determined on days 1, 2, 3, 4, 8 and 15. Macroscopic examination was performed at the end of the experimental period. 

No mortality occurred. 

Hunched posture and uncoordinated movements were observed in one female, which had recovered from the symptoms by day 4. 

The body weight gain shown by the animals over the study period was considered to be normal. 

No abnormalities were found at macroscopic past mortem examination of the animals. 

The minimum oral lethal dose of 1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE in rats was established as exceeding 2000 mg/kg body weight. 

Based on these results and according to the EEC criteria for classification and labelling requirements for dangerous substances and preparations (Guidelines in Commission Directive 93121/EEC, 27th April 1993), 1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE does not have to be classified and has no obligatory labelling requirement. 

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Study Monitor 

Testing Facility 

Study Oi rector 

study Plan 

TEST SUBSTANCE 

The Netherlands 

-------=---~ 

NOTOX B.V. 

Hambakenwetering 3 5231 DO 's-Hertogenbosch 

The Netherlands 

Start 

Pilot Study 4 December 1996 

Main Study : 10 December 1996 

End : 24 December 1996 

The sponsor is responsible for the completeness and GLP Compliance of all test 

substance data. 

Identification 

Chemical name 

CAS No. 

Description 

Batch 

Purity 

Test substance storage 

Stability under storage conditions 

Expiry date Density 

Preparation 

PURPOSE AND RATIONALE 

1,1,3,3-tetramethylbutyl peroxypivalate Propaneperoxic acid, 2,2-dimethyl-,1,1,3,3- tetramethylbutyl ester (9CI) 22288-41-4 

~olourless liquid 

(__ ' . See-eer·Joi-rtcate of Ana 1 ys is 

In freezer in the dark 

Stable 

01 June 1997 

0.87 g/ml (Determined at NOTOX) 

The test substance was dosed undiluted as delivered 

by the sponsor. 

The objective of this study was to assess at which maximum fixed dose level 1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE causes evident toxicity, but no 

mortality, when administered to rats as a single oral dose. This study should provide part of a rational basis for risk assessment in man. 

The oral route was selected as it is a possible route of human exposure during 

manufacture, handling or use of 1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE. 

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; 

, 

for the Determination of Toxicity, 81 bis: "Acute toxicity-oral, Fixed Dose Method". Official Journal of the European Communities, L383, December 1992. 

DECO "Guidelines for Testing of Chemicals", Section 4, Health Effects, No.420, "Acute Oral Toxicity- Fixed Dose Method", Paris Cedex, adopted 17 July, 1992. 

ARCHIVING 

NOTOX B.V. will archive the following data for at least 10 years: raw data, protocol, report and test substance reference sample. All magnetic media produced on this study will be maintained indefinitely. 

TEST SYSTEM 

Test System 

Age at start of Treatment 

Body weight at start 

. of treatment 

Total number of animals 

Identification 

ANIMAL HUSBANDRY 

Conditions 

Rat, Wistar strain Crl:(WI) BR (outbred, SPF-Quality) Recognised by international guidelines as the recommended test system (e.g. DECO, EEC). Source : Charles River, Sulzfeld, Germany. Approx. 6 weeks. 

Within +1- 20% of the sex mean. 

Pilot study: 1 female. Main study : 5 males and 5 females. Earmark 

Air-conditioned room with approximately 15 air changes per hour and the environment contra 11 ed wi. th optima 1 conditions considered as being a temperature of 21°C and a relative humidity of 50%. Fluctuations from these optimal conditions were noted, but were considered not to have affected study integrity. Lighting was 12 hours artificial fluorescent light and 12 hours dark per day. 

Accommodation 

Individual housing of animals in the Pilot study and group housing of five animals per sex per cage in the Main study in labelled polycarbonate cages containing purified sawdust as bedding material (Woody SPF, supplied by B.M.I., Helmond, The Netherlands). Certificates of analysis were examined and then retained in the NOTOX archives. 

Acclimatisation period was at least 5 days before start of treatment under laboratory conditions. 

Diet 

Free access to standard pelleted laboratory animal diet (from Carfil Quality BVBA, Oud-Turnhout, Belgium). Certificates of analysis are examined and then retained in the NOTOX archives. 

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Initially a "Pilot study", i.e. NOTOX Project 192847 was conducted in which one 

female rat was orally dosed with 1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE at 2000 mg/kg (2.3 ml/kg) body weight. No mortality occurred at this dose level. 

Based on the results, the main study was selected with one dose level and five 

male and five female animals dosed at 2000 mg/kg body weight. 

TREATMENT 

A health inspection was performed prior to commencement of treatment, to ensure 

that the animals were in a good state of health. 

Method 

Fasting 

Frequency 

Dose level/volume 

OBSERVATIONS 

Period Mortality/Viability 

Body weights 

Pilot study Main study 

Clinical signs (Pilot and main study) 

Necropsy (Main study) 

INTERPRETATION 

Oral gavage using stainless steel stomach tube. 

Food was withheld overnight prior to dosing until 

approximately 3-4 hours after administration of the test 

substance. 

Once, on day 1. 

Group 1: 2000 mg/kg (2.3 ml/kg) body weight. 

Dose volume calculated as follows: 

dose level (g/kg): density (g/ml) 

.Pilot study: 8 days. Main study: 15 days. 

Twice daily ·(Pilot and main study). 

Days 1 (pre-administration) and 8. 

Days 1 2, 3, 4, 8 and 15. 

At periodic intervals on the day of dosing (day 1) and 

once daily thereafter. The time of onset, degree and duration were recorded. 

All animals surviving to the end of the observation period were sacrificed by oxygen/carbon dioxide 

asphyxiation. All animals assigned to the study were subjected to necropsy and descriptions of all macroscopic 

abnormalities recorded. 

The results were evaluated according to the EEC criteria for classification and labelling requirements for dangerous substances and preparations (Guidelines in 

Commission Directive 93/21/EEC, 27th April 1993). 

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MAIN STUDY 

Mortality 

No mortality occurred in the animals during the study. 

Clinical Signs 

Hunched posture and uncoordinated movements were observed in one female, which had recovered from the symptoms by day 4. 

Body Weight 

The body weight gain shown by the animals over the study period was considered to be similar to that expected of normal untreated animals of the same age and strain. 

Macroscopic Findings 

No abnormalities were found at macroscopic post mortem examination of the animals. 

CONCLUSION 

The minimum oral lethal dose of 1,1,3,3-TETRAMETHYLBUTYL PEROXYPIVALATE in rats was established as exceeding 2000 mg/kg body weight. 

Based on these results and according to the EEC criteria for classification and labelling requirements for dangerous substances and preparations (Guidelines in Commission Directive 93/21/EEC, 27th April 1993), 1,1,3,3- TETRAMETHYLBUTYL PEROXYPIVALATE does not have to be classified and has no obligatory labelling requirement. 

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NUMBER OF DECEDENTS AFTER TREATMENT 

Test day 1 1 1 2 3 4 5 6 7 8 9 10 11 12 l3 14 15 Hours after treatment 0 2 4 Group 1 (2000 MG/KG) 

CLINICAL SIGNS 

FEMALES 

GROUP 1 (2000 MG/KG) 

Test day Time after treatment. Hours: 

ANlMAL 

NUMBER SIGNS 

1 NO CLINICAL SIGNS NOTED 

MAX. 

GRADE 

BODY WEIGHTS (GRAM) 

GROUP I SEX 

GROUP 1 I FEMALES 

(2000 MG/KG) 

ANIMAL 

1 

DAY 1 

145 

1 1 1 2 3 4 5 6 1 s 9 10 11 12 13 14 15 

0 2 4 

DAY S 

184 

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Test daY 

Time after treatment. Hours: 

ANIMAL 

NUMBER SIGNS 

1 NO CLINICAL SIGNS NOTED 

2 NO CLINICAL SIGNS NOTED 

3 NO CLINICAL SIGNS NOT~D 

4 NO CLINICAL SIGNS NOTED 

5 NO CliNICAL SIGNS NOTED 

FEMALES 

MAX. 

GRADE 

1 1 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 

0 2 4 

GROUP 1 (2000 MG/KG) 

Test day . Time after treatment. Hours: 

ANIMAL 

NUMBER SIGNS 

6 NO CLINICAL SIGNS NOTED 

7 NO CLINICAL SIGNS NOTED 

8 NO CLINICAL SIGNS NOTED 

9 NO CLINICAL SIGNS NOTED 

10 POSTURE 

MAX. 

GRADE 

1 1 1 2 3 4 s 6 7 8 9 10 11 12 13 14 15 

0 2 4 

HUNCHED POSTURE, ........... (1) - - l 1 1 

1 -

GAIT I MOTILITY 

UNCOORDINATED MOVEMENTS.,,. (3) 

- ~ sign not observed I • = observation not performed I + ~ animal dead 

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4 170 188 204 213 255 318 

5 168 179 195 206 245 315 

MEAN 169 181 197 207 246 306 

ST.OEV. 3 5 5 4 6 10 

N ; 5 5 5 5 5 

GROUP 1 I FEMALES 6 1% 148 !58. 162 183 212 

(2000 MG/KG) 7 144 153 164 170 183 204 

8 148 157 171 176 197 211 

9 138 146 !64 17-1 !S9 201 

10 141 147 !50 160 189 217 

MEAN 141 150 161 168 188 209 

ST.OEV. 5 5 8 7 6 6 

N 5 5 5 5 5 5 

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NO FINDINGS NOTED 

ANIMAL 2 

NO FINDINGS NOTED 

ANIMAL l' 

NO FINDINGS NOTED 

ANIMAL 4 

NO FINDINGS NOTED 

ANII~AL 5 

NO FINDINGS NOTED 

FEMALES 

GROUP 1 (2000 MG/KG) 

ANIMAL 6 

NO FINDINGS NOTED 

ANIMAL 7 

NO FINDINGS NOTED 

ANIMAL 8 

NO FINDINGS NOTED 

ANIMAL 9 

NO FINDINGS NOTED 

ANIMAL 10 

NO FINDINGS NOTED 

(SCHEDULED NECROPSY, 24-DEC-96, DAY 15 AFTER TREATMENT) 

(SCHEDUlED NECROPSY, 24-0EC~96, DAY 15 AFTER TREATMENT) 

(SCHEDULED NECROPSY, 24-DEC-96, DAY 15 AFIER TREATMENT) 

(SCHEDULED NECROPSY, 24-DEC-96, DAY 15 AFTER TREATMENT) 

(SCHEDULED NECROPSY 1 24-DEC-96 1 DAY 15 AFTER TREATMENT) 

{SCHEOULEO NECROPSY, 24-DEC-96, DAY 15 AFIER TREATMENT) 

(SCHEDULED NECROPSY, 24-DEC-96, DAY 15 AFTER TREATMENT) 

(SCHEDULED NECROPSY, 24-DEC-96, DAY 15 AFTER TREATMENT) 

(SCHEDULED NECROPSY, 24-DEC-96, DAY 15 AFTER TREATMENT) 

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TEST SUBSTANCE CERTIFICATE OF ANALYSIS 

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ReffCS-331 

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Product nama 

i:'f;t~e _nama 

B~!chnumbar 

: 1,1 ,3,3-tetramethylbutyl peroxypiva!ate 

:-Ttig.QllilX 425 C75 

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Ari~IYtfcal 

documanta~c>"n · 

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Fii~cad~ 

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Autfiorlzad bv---~-----------­

Na(na 

Function 

D~ta 

Signature 

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