Document 4aEJgykrQLBD4k4ZG8x69m3NQ
STUDY CODE : K01-1802
A R 1 2 S -O I3 C
Receipt No. T96-2481 Report No. T-4637
FINAL REPORT
BACTERIAL REVERSE MUTATION TEST OF T -l
September, 1996
Hita Research Laboratories Chemical Biotesting Center Chemicals Inspection & Testing Institute
Japan
002000
QUALITY ASSURANCE STATEMENT
Hita Research Laboratories, Chemical Biotesting Center Chemicals Inspection & Testing Institute, Japan
Sponsor:
SUMITOMO 3M LIMITED
Title:_______ Bacterial reverse mutation test of z -1
Study code: K01-1802
This report was audited by the Quality Assurance Section. I, the undersigned, hereby declare that this report reflects the original Japanese report.
(Date)
(Signature) Section Chief, Quality Assurance
02001
KOl-1802
I, the undersigned, hereby declare that this report provides a correct English translation of the Final Report. (Study code No. KOI-1802 issued on September 25, 1996)
(date)
/?9 7
('signature') Shozo Ogura
Hita Research Laboratories Chemical Biotesting Center Chemicals Inspection & Testing Institute, Japan
CG2002
K01-1802
GLP STATEMENT
Hita Research Laboratories, Chemical Biotesting Center Chemicals Inspection & Testing Institute, Japan
Sponsor:___________ SUMITOMO 3M LIMITED Title:_______________Bacterial reverse mutation test of T - l Study Code No : K01-1802
I, the undersigned, hereby declare that this study was conducted in compliances with "Standards to be observed by Testing Institutions for Toxicity Investigations" (Japan's MOL, No.76, September 1, 1988).
Management:
Signed in original
September 25, 1996
Shigetaka Yamane, Ph. D.
002003
GLP STATEMENT
Hita Research Laboratories, Chemical Biotesting Center Chemicals Inspection & Testing Institute, Japan
Sponsor:___________ SUMITOMO 3M LIMITED Title: _________ Bacterial reverse mutation test of T -1 Study Code No.:_____K01-1802
I, the undersigned, hereby declare that this study was conducted in compliances with "Standards to be observed by Testing Institutions for Toxicity Investigations" (Japan's MOL, No.76, September 1, 1988).
This was reissued because of the final report amendment.
Management:
Signed in original Shigetaka Yamane, Ph. D.
November 27,1996
002004
KOl-1802
QUALITY ASSURANCE STATEMENT
Hita Research Laboratories, Chemical Biotesting Center Chemicals Inspection & Testing Institute, Japan
Sponsor:___________ SUMITOMO 3M LIMITED Title:______________ Bacterial reverse mutation test of T -1 Study Code No : K01-1802
This study was audited by the Quality Assurance Section and the study procedures were inspected on the following dates.
Dates of Inspections
Dates of Reports to
Dates of Reports to
and Audits
Study Director
Management
August 15, 1996
August 15, 1996
August 15, 1996
September 20,1996
September 24, 1996
September 24, 1996
September 25, 1996
September 25, 1996
September 25, 1996
I, the undersigned, hereby declare that this report provides an accurate description of the
methods and procedures used in this study and that the reported results accurately reflect
the raw data obtained.
Section Chief, Quality Assurance:
Signed in original Keiji Shiraishi, B.S.
September 25, 1996
CG200S
QUALITY ASSURANCE STATEMENT
Hita Research Laboratories, Chemical Biotesting Center Chemicals Inspection & Testing Institute, Japan
Sponsor:___________ SUMITOMO 3M LIMITED
Title:______________ Bacterial reverse mutation test of T - l
Study Code No.:
KO1-1802
This study was audited by the Quality Assurance Section on the following dates.
Auditor Kumiko Matsui
Date of Audit
Date of Report to Study Director
Date of Report to Management
November 27, 1996 November 27, 1996 November 27, 1996
This statement was added to the Quality Assurance Statement issued on September 25, 1996.
Section Chief, Quality Assurance:
Signed in original Keiji Shiraishi, B.S.
November 27, 1996
\
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Study code:
K01-1802
Test substance code: HR3272
Sponsor code:
S-030
TITLE Bacterial reverse mutation test of T - l
SPONSOR SUMITOMO 3M LIMITED 8-8, Minami-Hashimoto 3-chme Sagamihara-shi, Kanagawa, 229 Japan
TESTING FACILITY Hita Research Laboratories, Chemical Biotesting Center Chemicals Inspection & Testing Institute, Japan 822, 3-chome, Ishii-machi, Hita, Oita 877, Japan
PURPOSE OF STUDY The purpose of this study was to determine the mutagenic potential o f the test substance using Salmonella typhimurium and Escherichia coli.
TESTING METHOD This study was conducted in accordance with the following guidelines: "Standards for Toxicity Investigations" (Japan's MOL, No.77, September 1, 1988).
GLP COMPLIANCE This study was carried out in compliance with the following GLP requirement: "Standards to be observed by Testing Institutions for Toxicity Investigations" (Japan's MOL, No.76, September 1, 1988).
PERIOD OF STUDY Commencement of test: Dose finding test: Completion of observation: Presentation of final report:
August 21, 1996 September 10, 1996 September 17, 1996 September 25, 1996
CG2007
KOl-1802
LOCATION AND PERIOD FOR RETENTION OF RAW DATA Data and test substance are retained in the archives and the test substance storage room of Hita Research Laboratories for 10 years following the date o f the notification specified under Item 1 of Article 57-2 of Industrial Safety & Health Law, respectively.
After termination of the retention period, any measures taken are done so with the approval of the sponsor.
PERSON CONCERNED WITH STUDY
Study Director:
Signed in original
September 25, 1996
Shozo Ogura
Hita Research Laboratories
Mutagenicity Section
Study Staff: Person in charge of Storage:
Tsunehiko Inai, B.S. Shizuka Kouda
ANY UNEXPECTED SITUATIONS AND DEVIATIONS FROM PROTOCOL There were no unexpected situations and deviations from protocol which might have affected the test results.
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CONTENTS
SUMMARY
MATERIALS AND METHODS
1. TEST SUBSTANCE AND POSITIVE CONTROLS
............................
2. BACTERIAL STRAINS
3. MEDIUM AND S9 MIX
4. PRE-CULTURES
....................................................
5. PREPARATION OF TEST SUBSTANCE AND POSITIVE
CONTROLS
....................................................
6. METHODS
....................................................
7. MICROSCOPIC OBSERVATION AND COLONY COUNTING .......
8. INTERPRETATION OF RESULTS ....................................................
2
6
7 7 8 9
RESULTS
.................................................... 9
CONCLUSION
......................... : .........................
10
REFERENCES
..................................... .............. 10
APPENDIX TABLES AND FIGURES .................................................... 1-6
1
4 5
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SUMMARY
The reverse mutation test o f T - l was performed on Salmonella typhimurium strains TA100, TA1535, TA98, TA1537 and Escherichia coli strain WP2 uvrA using the pre incubation method with and without metabolic activation. The results showed that the numbers of their revertant colonies for all strains in groups which were treated with the test substance were less than twice that o f each negative control, with and without S9 Mix. The numbers of the revertant colonies in the negative control and the positive controls were within the background data in our laboratories. Based upon the above results, T - l was judged to have no reverse mutagenic potential under the present test conditions.
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KOI-1802
MATERIALS AND METHODS
1. TEST SUBSTANCE AND POSITIVE CONTROLS 1.1 Test substance (Information provided by the sponsor)
1) Name Reaction products of perfluorodimethylcyclohexylsulfonyl fluoride, perfluoroalkyl (C=0-2) cyclohexylsulfonyl fluoride, potassium carbonate and sulfuric acid Other name: T - l CAS No.: 67584-42-3 (the main component A(n=2))
2) Lot No. 293
3) Supplier SUMITOMO 3M LIMITED
4) Structural formula or rational formula (Outline of manufacturing method, in case both were unknown)
(n=0-2)
76%
C CmF2mfiS03K
(m=8)
D K2S 04
Others (molecular formula --)
20%
3%
0.4% 0.6%
100 w/w% 6) Impurities
7) Physicochemical properties
Appearance at ordinary temperature: white powder
Molecular weight:
about 500
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K01-1802
Stability:
stable
Melting point:
Boiling point:
Vapor pressure:
Partition coefficient:
Solubility:
Degree of solubility:
Water:
S 0.1%
DMSO:
^ 5 w/v%*
Acetone: about 5%
Others:
methanol about 5%
* Examined in our laboratories
8) Storage conditions
room temperature
9) Care on handling
Gloves, a mask, a head cap and a lab coat were worn when handling.
1.2 Positive controls
1) 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (AF-2)
Manufacturer: Wako Pure Chemical Industries, Ltd.
Lot No.:
LEN0571
Properties:
reddish-orange crystalline powder
Purity:
99.5%
Grade:
special grade
2) Sodium azide (NaN3)
Manufacturer: Wako Pure Chemical Industries, Ltd.
Lot No.:
DLP2438
Properties: white crystalline
Purity:
99.4%
Grade:
special grade
3) 2-Methoxy-6-chloro-9-[3-(2-chloroethyl)-aminopropylamino]acridine 2HC1
(ICR-191)
Manufacturer: Polysciences, Inc.
Lot No.:
412795
Properties:
yellow crystalline powder
Purity:
--
Grade:
--
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4) 2-Aminoanthracene (2AA)
Manufacturer: Wako Pure Chemical Industries, Ltd.
Lot No.:
DLR7869
Properties: yellowish-green-brown powder
Purity:
95.7%
Grade:
--
5) Storage conditions
A cold and dark place
6) Care on handling
Gloves, a mask, a head cap and a lab coat were worn when handling.
2. BACTERIAL STRAINS 2.1 Strains selected
Salmonella typhimurium strains TA100, TA98, TA1535 and TA1537 were obtained from Dr. B.N. Ames, University o f California, U.S.A. ,on June 20, 1990.
A Escherichia coli strain WP2 uvrA was obtained from Japan Bioassay Laboratories, on April 6, 1995.
S. typhimurium strains TA100, TA1535 and E. coli strain WP2 uvrA were used for the detection of base-pair substitution mutation, while S. typhimurium strains TA98 and TA1537 for the detection of frameshift mutation. 2.2 Storage The test strains were stored as frozen stock cultures (0.045 ml of dimethyl sulfoxide (DMSO)*/0.5 ml o f broth culture) at -80C (ultra-deep freezer MDF291, Sanyo). * Purity ^ 99.0%, Lot No. CF103, Dojindo Laboratories
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2.3 Characterization of the strains 1) Characteristics of the strains__________________________________ _
Mutation on Mutation on Membrane R-factor
Strains
synthesis of excision mutation (pKMIOl)
_______________________amino acid repair_______(LPS)______________
Salmonella typhimurium
TA1535
hisG46
AuvrB
rfa
-
TA1537 TA98 TA100
hisC3076 AuvrB rfa -
hisD3052 AuvrB rfa +
hisG46
AuvrB
rfa
+
Escherichia coli
WP2 uvrA
trp
AuvrA
+
The amino acid requirement for growth was demonstrated by using histidine for
S. typhimurium strains and tryptophan for E. coli strain. The presence of R-
factor, membrane mutation and mutation on the ability to repair DNA lesions were
confirmed by ampicillin resistance, sensitivity to crystal violet and UV sensitivity,
respectively.
2) Date of characterization_____________________________________________
Salmonella typhimurium
TA1535
July 18, 1996
TA1537
June 7, 1996
TA98
June 7, 1996
TA100
March 6, 1996
Escherichia coli
WP2 uvrA
April 18, 1996
3. MEDIUM AND S9 MIX 3.1 Medium
1) Minimal glucose agar plate (prepared in our Laboratories)
Components
Amount included in one litre
20 x Vogel-Bonner E
50 ml
40 w/v% Glucose
50 ml
Agar
15 g
(1) Agar:
Bacto-Agar (Lot No. 84707AJA, Difco Laboratories)
(2) Manufacturing date: dose finding test on August 29, 1996
main test on September 5, 1996
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2) Soft agar
The solution containing 0.5 mM histidine and 0.5 mM biotin for S. typhimurium
strains or 0.5 mM tryptophan for E. coli strain was added to the soft agar
solution containing 0.6 w/v% agar (Bacto-Agar, Lot No. 71892AJB, Difco
Laboratories) and 0.5 w/v% NaCl in a ratio o f 1 : 10.
3.2 S9 Mix
1) Rat liver S9 (Kikkoman Co., Ltd.)
Induction method:
SD male rats, 7-week-old (203-254 g), were
intraperitoneally administrated phnobarbital (30 mg/kg
x 1 time, 60 mg/kg x 3 times) and 5,6-benzoflavone (80 mg/kg x 1 time).
Lot No.:
RAA-350 (manufactured on August 23, 1996,
Storage:
purchased on September 4, 1996) -80^3 (ultra-deep freezer MDF-291, Sanyo)
2) Cofactor for S9 Mix (Oriental Yeast Industries, Ltd.)
Lot No.:
999601
Storage:
-20C (bio-freezer GS-2603, Nippon Freezer Ltd.)
3) Composition of S9 Mix
One ml of S9 Mix contained 8 pmol MgCL, 33 pmol KC1, 5 pmol G-6-P, 4 pmol NADPH, 4|imol NADH, 100 pmol of 0.2 M sodium-phosphate buffer (pH 7.4) and 0.1 ml S9.
4. PRE-CULTURES From the stock cultures, 20 pi of bacterial suspension was inoculated to L-tube containing 10 ml nutrient broth No.2 (Lot No. 194 56443, OXOID Ltd.), the bacterial culture was incubated at 37 0.5C for 8 h with shaking at 50 times/min by the Monod shaker (MONOSIN- II A, Taitec Co., Ltd.) The viable cell counts calculated from the values which were determined at 660 nm by spectrophotometry (Novaspec, LKB Japan) at the end of incubation are shown below.
No. o f Dose viable cells finding test (x l0 9/ml) Main test
TA100 1.9 1.9
TA1535 WP2 uvrA 2.2 3.4 2.1 3.6
TA98 2.4 2.3
TA1537 2.1 2.0
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5. PREPARATION OF TEST SUBSTANCE AND POSITIVE CONTROLS 5.1 Test substance
1) Preparation The test substance was dissolved in DMSO (Lot No. CF103) to make 5 w/v% concentration and diluted with the same solvent to give appropriate concentrations.
2) Stability of the test solution No denaturation of the test solution was observed to a color and exothermic reaction until 2 hours after preparation.
3) Preparation time Prepared immediately before use and used within 0.5 h at room temperature.
5.2 Positive controls 1) Preparation NaN3 was dissolved in distilled water (Distilled water for injection, Lot No. K6B74, Otsuka Pharmaceutical Factory). AF-2, ICR-191 and 2AA were dissolved in DMSO (Lot No. CD069). 2) Preparation time and storage condition Prepared on every 3 months and stored at -80C (ultra-deep freezer MDF-291, Sanyo).
6. METHODS The test was carried out for S. typhimurium strains TA1535, TA1537, TA98, TA100 and a E. coli strain WP2 uvrA using the pre-incubation method both with and without metabolic activation system. The plating was done in triplicate for the negative control and in duplicate for the test substance and positive controls.
6.1 Procedures After 0.1 ml of the test substance solution, 0.5 ml of 0.1 M sodium phosphate buffer (pH 7.4) or S9 Mix, and 0.1 ml o f the bacterial culture were added to a tube, the mixtures were incubated for 20 min at 37 0.5C. Two ml of the soft agar was then added to each tube and poured onto a minimal glucose agar plate. After incubation for 48 h at 37 0.5C, the number o f revertant colonies were counted.
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As the sterility test, each 0.1 ml of each bacterial suspension, test substance solution, S9 Mix or 0.1 M sodium phosphate buffer (pH 7.4) were smeared on a minimal glucose agar plate and incubated at 37 0.5C for 48 h, and then checked the bacterial contamination. DMSO was used as a negative control, and the following positive controls were used for each bacterial strains.
S9 Mix (-) S9 Mix (+)
TA100 AF-2 0.01 2AA
1
TA1535 NaN3 0.5 2AA 2
WP2 uvrA AF-2 0.01 2AA 10
TA98 AF-2 0.1 2AA 0.5
TA1537 ICR-191
1 2AA
2
(pg/plate) 6.2 Dose selection
1) Dose finding test The test was carried out at the highest dose of 5,000 pg/plate and 6 doses of 1,000, 500, 100, 50, 10 and 5 pg/plate. As a result, growth inhibition observed at doses of more the 1,000 pg/plate for S. typhimurium strains TA100, TA1535, TA1537 and E. coli strain WP2 uvrA and 5,000 pg/plate for S. typhimurium strain TA98 without S9 Mix. Growth inhibition observed at dose of 5,000 pg/plate for S. typhimurium strains TA100, TA1535, TA1537 a n d coli WP2 uvrA with S9 Mix.
2) Main test Based on the results of the dose finding test, a main test with S9 Mix was performed at the highest dose of 5,000 pg/plate and 5 lower doses diluted with a geometric progression of 2. Without S9 Mix, a maximum dose was decided 1,250 pg/plate in the case of S. typhimurium strains TA100, TA1535, TA1537 and E. coli strain WP2 uvrA and 5,000 pg/plate in the case of S. typhimurium strain TA98 and 5 lower doses were decided in each bacterial strain by dilution with a geometric progression of 2.
7. MICROSCOPIC OBSERVATION AND COLONY COUNTING 7.1 Microscopic observation
The state of revertant colonies (size and number of colonies), deposition of the test substance and the growth inhibition were examined with a stereo microscope.
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7.2 Colony counting The number of colonies were counted with a manual counter or a colony analyzer (CA-9, Toyo-sokki Co., Ltd). Correction for counting errors was made for measurements with the colony analyzer. Each plate was measured three times, and the average of these three measurements was adopted as the number of revertant colonies on the plate. The average for each dose was calculated from the values of the plates used. Decimals of the average figures were rounded off.
8. INTERPRETATION OF RESULTS The test substance was judged to be positive, when the number of revertant colonies was twice or more of the negative control, and the dose-relationship and the reproducibility were obtained. Any statistical procedures were not used.
RESULTS
The numbers of their revertant colonies for all strains in groups which were treated with the test substance were less than tw ee that of each negative control with and without S9Mix.
The growth inhibition was observed at more than 1,000 pg/plate in S. typhimurium strains TA100, TA1535, TA1537 and. coli strain WP2 uvrA and at 5,000 pg/plate in S. typhimurium strain TA98 without S9 Mix. The growth inhibition was noted at 5,000 pg/plate in S. typhimurium strains TA100, TA1535, TA1537 and E. coli strain WP2 uvrA with S9 Mix.
The positive controls showed the distinct increase of revertant colonies, and the positive controls and the negative control were within a range o f the background data in our laboratories.
There were no fluctuations which affected the test results since the sterility test confirmed the absence of any micro-organisms.
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CONCLUSION
In conclusion, T - l was judged to have no reverse mutagenic potential under the present test conditions.
REFERENCES
1. Ministry of Labor (1991) Guidebook on Mutagenicity Tests using Micro organisms, New Edition (in Japanese) published by Japan Industrial Safety and Health Association,
2. Green M.H.L. and W.J. Muriel (1976) Mutagen testing using Trp+ reversion in Escherichia coli, Mutation Res., 38: 3-32.
3. Maron, D.M., and B.N. Ames (1983) Revised methods for the Salmonella mutagenicity test, Mutation Res., 113: 173-215.
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Dose finding test
KOl-1802
Test substance: r --1
With(+) or without(-)
S9 Mix
Test substance concentration
(ug/plate)
negative control
Number of revertants (number o: colonies/plate)
Base-pair substitution type
Framesh] ft type
TA 100
104 97 ( 98) 94
TA 1535 9 9 ( ID 16
WP2tfwii
19 21 ( 20) 21
TA 98
20 25 ( 23) 23
TA 1537
12
14 ( 13) 13
S9 Mix (-)
5 * 98> M6 ( 10) 19 < 22> u { 21) 9 ) 10 u ! * io) 16 1 "> S < z) 22 < 2) 2 i n)
50 2 < 94) 10 < 10> 3 ( 18) 28 ZZ) 12 ( 9)
100 S < 85) u 1 111 3 ( 2i) 3 ( 21) 12 ( ID
500 2 ( 95)
" ( 17) 2 < 245 " t lfl)
1000 < 5*) p i8*) 3 < 21) i 10*)
5000 negative control
<- 85*)
94 94 ( 98) 107
3 < *) 17 9 ( ID 7
m i 16*) 29 24 ( 29) 33
! f . ( is>) S i *)
34 20 30 ( 35) 15 ( 18) 42 18
S9. M i x (+)
5 94 < 95) " < 12) " < 35 i 37) u t 17>
10 85 < 9) " t ^ 3 i Z9) 3 i Z9) p i 19)
50 S < 85> S ( 11] 23 < 27> 'S < ) 2 ( i4)
100 2 ( ^
1! <
3 < 29) a
500 3 < 9?) " t io) 3 < Z5> 40 (. 3Z) 22 < 18)
1000 105 < "> " i a) 5 ( zfi) 38 ( 34) 12 1 111
5000 S * 90*) 7*)
> S i 29) 3< 6*)
Positive control not requiring
S9 Mix
Name AF-2
Concentration (ug/plate)
0.01
Number of
colonies/plate S <*)
NaNa 0.5
290 i 302)
AF-2 0.01
157 <147)
AF-2 ICR-191 0.1 1
S! i ) 3 S i2417)
Positive control requiring S9 Mix
Name
Concentration (ug/plate) Number of
colonies/plate
2AA 1
m t 666)
2AA 2
S ( i)
Notes Parenthesis shows the mean of each plate.
* : Observed bacterial growth inhibition.
AF-2: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide
2AA 10
S ( fi" )
2AA 0.5
m t 287)
2AA 2
UR t 1" )
NaN3: Sodium azide
ICR-191: 2-Methoxy-6-chloro-9-(3-(2-chloroethyl)-aminopropylamino) acridine-2HCl
2AA: 2-Aminoanthracene
CO2020
1
Main test
Test substance: t -- 1
K01-1802
Nith(t) oi without(-)
S9 Mix
Test substance concentration
(lig/plate)
negative control
Number of revertants (number o colonies/plate)
Base-pair substitution type
Framesh: ft type
TA 100 TA 1535 WP2yrri TA 98 TA 1537 93 6 21 26 9 84 ( 94) 14 ( 10) 18 ( 21) 28 ( 27) 9 ( 10) 105 11 25 28 11
39.1 S < 90> ( 14) 5 < l9>
"i
78.1 }5 ( 112) S ( 16> S<
12 ( 10)
S9 M ix (-)
156 B < 97> ( 17> ( " > S ( 21) n < 15> 313 B 19 ( 10) t 2j 29 t 2?) 11 " )
625 S t ) " ( 9) S a > S t 23) ( 9>
1250 < 91*>
16*> S t 18*) a < W St 8*)
2500 23 ( 22)
5000 negative control
102 84 ( 98) 109
8 8 ( 8) 8
31 30 ( 33) 37
S t 18*) 36 21 38 ( 37) 20 ( 22) 38 24
156 m <103> 1 < 1) S t ) S ( 34> 1 2D
S9 M ix (+)
313 U3 ( 101> 11 ( 9) 27 t 89) S ( 30) S t 18)
625 106 t 97> < " 1 S t 33) S t 31) S ( 14)
1250 92 < 86> ( 101 " t BJ 1 " ) 2t 2D
2500 m ( 97) !< 16 t *> S t 30) 5! 1 15>
Positive control not requiring
S9 Mix
5000
Name Concentration
(ug/plate) Number of colonies/plate
SJ* -
AF-2
0.01
m (W
5< *> 30*t 36*) <
NaNa AF-2 AF-2
0.5 0.01 0.1
, S t io*> ICR-191
1
387 ( 37) iS t m S! 579> SS <1886>
Positive control requiring S9 Mix
Name Concentration
2AA
1
2AA
2
dumber of
coloniem/plate m <728> m ( 127)
Notes Parenthesis shows the mean of each plate.
* : Observed bacterial growth inhibition.
AF-2: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide
2AA
10 S! t 861)
2AA
0.5 S t 27)
2AA
2
1I5543 (( i1m54i)
NaNa: Sodium azide
ICR-191: 2-Methoxy-6-chloro-9-(3-(2-chloroethyl)-aminopropylamino) acridine*2HC1
2AA: 2-Amirioanthracene
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Revertant colonies per plate
Revertant colonies per plate
KOI--1802
Dose finding test
Dose finding: test
K01-1802
Revertant colonies per plate
Revertant colonies per plate
0:TA 98 :TA 1537
Fig. 4 Dose-re s p o n s e curve wit h S9 Mix
~ 4-
(g/plate)
02023
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02024
Revertant colonies per plate
Revertant colonies per plate
K01-1802
Main test
--6--
C02025
Revertant colonies per plate
Revertant colonies per plate
M a i n test
K01-1802
FINAL REPORT AMENDMENT
Hita Research Laboratories, Chemical Biotesting Center Chemicals Inspection & Testing Institute, Japan
1. Title (Study code) Bacterial reverse mutation test of T - l (K01-1802)
2. Amendment (Items) CAS No. and structural formula or rational formula (Annex 1)
3. Authorization Study Director
Signed in original Shozo Ogura
November 27. 1996
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Annex 1
Before changes: 1)
(Information provided by the sponsor) Page 2 Name Reaction products of perfluorodimethylcyclohexylsulfonyl fluoride,perfluoroalkyl (C=0-2) cyclohexylsulfonyl fluoride, potassium carbonate and sulfuric acid Other name: T-1 CAS No.: 67584-42-3
4) Structural formula or rational formula (Outline o f manufacturing method, in case both were unknown)
CnF2n+l
F )-- S 0 3K
(n=0-2)
CmF2mHS03K
(m=8)
K2S 0 4
(molecular formula --)
-2 -
After changes: 1)
Name Reaction products of perfluorodimethylcyclohexylsulfonyl fluoride,perfluoroalkyl (C=0-2) cyclohexylsulfonyl fluoride, potassium carbonate and sulfuric acid Other name: T - l CAS No.: 67584-42-3 (the main component A(n=2i)
4) Structural formula or rational formula (Outline of manufacturing method, in case both were unknown)
A CnF2n+l
B CF3
s o 3k
cf3
C CmF2mflS03K
(m=8)
D K2SO4
Others (molecular formula
--)
76%
20% 0.4%
M%
Reason for changes: To document new information supplied by the sponsor.
Date effective:
November 13,1996
- 3 - C-03028
