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EC number: 200-677-4 | CAS number: 68-11-1
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![](https://echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 003
- Report date:
- 2003
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- Ammonium mercaptoacetate
- EC Number:
- 226-540-9
- EC Name:
- Ammonium mercaptoacetate
- Cas Number:
- 5421-46-5
- Molecular formula:
- C2H4O2S.H3N
- IUPAC Name:
- ammonium sulfanylacetate
- Details on test material:
- Ammonium Thioglycolate 71%
Source: Bruno Bock Chemische Fabrik GmbH & Co KG
Description: colourless liquid
Batch number: 4804
Content of active ingredient: 71.2%
Constituent 1
Method
- Target gene:
- Histidine reversion
Species / strain
- Species / strain / cell type:
- S. typhimurium, other: TA1535, TA1537, TA98, TA100 and TA102
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-mix (from livers of male Sprague-Dawley rats treated by Aroclor 1254)
- Test concentrations with justification for top dose:
- 15, 50, 150, 500, 1500 and 5000 µg a.i./plate.
Two distinct experiments were performed using these doses. - Vehicle / solvent:
- sterile distilled water
Controls
- Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- 9-aminoacridine
- N-ethyl-N-nitro-N-nitrosoguanidine
- benzo(a)pyrene
- mitomycin C
- other: 2-aminoanthracene, 1,8-Dihydroxyanthraquinone
- Details on test system and experimental conditions:
- METHOD DETAILS:
- Test concentrations:
. Preliminary study (on TA100): 0, 0.15, 0.5, 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate
. Main study: see the "Test concentration" field
- Number of replicates: 3
- Positive controls:
without S9-mix
. for TA1535 and TA100: N-ethyl-N'-nitro-N-nitrosoguanidine (5 and 2 µg/plate respectively)
. for TA1537: 9-aminoacridine (80 µg/plate)
. for TA98: 4-Nitroquinoline-1-oxide (0.2 µg/plate)
. for TA102: mitomycin C (0.5 µg/plate)
with S9-mix
. for TA1535, TA1357 and TA100: 2-aminoanthracene (2; 2 and 1µg/plate respectively)
. for TA98: benzo(a)pyrene (5 µg/plate)
. for TA102: 1,8-Dihydroxyanthraquinone (10 µg/plate)
- Pre-incubation time: no
- Pre-incubation temperature: no
- Incubation time: 48h
- Incubation temperature: 37°C
EXAMINATION:
- Bacterial toxicity: determined by examination of background lawn growth
- Number of revertants / plate
ANALYTICAL DEVICE: Colonies were counted electronically using a Domino Colony Counter. - Evaluation criteria:
- The reverse mutation assay may be considered valid if the following criteria are met: All tester strain cultures exhibit a characteristic number of spontaneous revertants per plate in the vehicle and untreated controls. Acceptable ranges are presented below with historical control ranges for 2001 and 2002 presented in Table 1.
Spontaneous Mutation Ranges:
TA1535 7 to 40
TA100 60 to 200
TA1537 2 to 30
TA98 8 to 60
TA102 180 to 400
The test material may be considered positive in this test system if the following criteria are met: The test material should have induced a reproducible, dose-related and statistically significant increase in the revertant count in at least one strain of bacteria. - Statistics:
- Dunnett's method of linear regression
Results and discussion
Test resultsopen allclose all
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity, but tested up to precipitating concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- = 5000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 102
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- CYTOTOXICITY:
The test substance was toxic at 5000 µg/plate in the TA100 strain (without S9 mix: very weak bacterial background lawn, with S9 mix: sparse bacterial background lawn).
GENOTOXICITY:
No significant increases in the number of revertants were observed at any dose level, with or without metabolic activation.
Any other information on results incl. tables
Table 1:Spontaneous Mutation Rates (Concurrent Negative Controls)
EXPERIMENT 1
Number of revertants (mean number of colonies per plate) |
|||||||||
Base-pair substitution type |
Frameshift type |
||||||||
TA100 |
TA1535 |
TA102 |
TA98 |
TA1537 |
|||||
136 |
9 |
351 |
15 |
5 |
|||||
139 |
(135) |
14 |
(10) |
350 |
(358) |
18 |
(15) |
5 |
(6) |
129 |
7 |
373 |
13 |
8 |
EXPERIMENT 2
Number of revertants (mean number of colonies per plate) |
|||||||||
Base-pair substitution type |
Frameshift type |
||||||||
TA100 |
TA1535 |
TA102 |
TA98 |
TA1537 |
|||||
62 |
18 |
267 |
12 |
6 |
|||||
97 |
(73) |
11 |
(14) |
318 |
(301) |
12 |
(12) |
5 |
(5) |
60 |
13 |
318 |
12 |
5 |
Table 2: Test Results: Experiment 1 – Without Metabolic Activation
Test Period |
From : 25 March 2003 |
To : 28 March 2003 |
||||||||||
With or without S9-Mix |
Test substance concentration (µg/plate) |
Number of revertants (mean number of colonies per plate) |
||||||||||
Base-pair substitution type |
Frameshift type |
|||||||||||
TA100 |
TA1535 |
TA102 |
TA98 |
TA1537 |
||||||||
- |
0 |
153 121 131 |
(135) 16.4# |
9 7 8 |
(8) 1.0 |
367 310 385 |
(354) 39.2 |
13 16 24 |
(18) 5.7 |
6 7 11 |
(8) 2.6 |
|
- |
15 |
128 114 130 |
(124) 8.7 |
6 9 5 |
(7) 2.1 |
353 296 434 |
(361) 69.3 |
16 17 17 |
(17) 0.6 |
5 5 3 |
(4) 1.2 |
|
- |
50 |
104 129 118 |
(117) 12.5 |
12 12 5 |
(10) 4.0 |
399 360 399 |
(386) 22.5 |
16 14 12 |
(14) 2.0 |
5 6 4 |
(5) 1.0 |
|
- |
150 |
144 113 118 |
(125) 16.6 |
11 8 8 |
(9) 1.7 |
406 372 360 |
(379) 23.9 |
15 7 16 |
(13) 4.9 |
4 5 6 |
(5) 1.0 |
|
- |
500 |
101 106 118 |
(108) 8.7 |
11 13 11 |
(12) 1.2 |
362 359 393 |
(371) 18.8 |
15 8 12 |
(12) 3.5 |
5 3 3 |
(4) 1.2 |
|
- |
1500 |
116 104 106 |
(109) 6.4 |
5 3 11 |
(6) 4.2 |
382 356 368 |
(369) 13.0 |
17 15 18 |
(17) 1.5 |
7 7 4 |
(6) 1.7 |
|
- |
5000 |
0 V 0 V 0 V |
(0) 0.0 |
3 S 9 S 5 S |
(6) 3.1 |
204 221 153 |
(193) 35.4 |
17 S 14 S 13 S |
(15) 2.1 |
0 S 0 S 4 S |
(1) 2.3 |
|
Positive controls S9-Mix - |
Name Concentration (µg/plate) No. colonies per plate |
ENNG |
ENNG |
MMC |
4NQO |
9AA |
||||||
3 |
5 |
0.5 |
0.2 |
80 |
||||||||
553 474 505 |
(511) 39.8 |
287 284 256 |
(276) 17.1 |
862 859 903 |
(875) 24.6 |
75 81 80 |
(79) 3.2 |
2605 2689 2688 |
(2661) 48.2 |
|||
Table 3: Test Results: Experiment 1 – With Metabolic Activation
Test Period |
From : 25 March 2003 |
To : 28 March 2003 |
||||||||||
With or without S9-Mix |
Test substance concentration (µg/plate) |
Number of revertants (mean number of colonies per plate) |
||||||||||
Base-pair substitution type |
Frameshift type |
|||||||||||
TA100 |
TA1535 |
TA102 |
TA98 |
TA1537 |
||||||||
+ |
0 |
173 145 180 |
(166) 18.5# |
16 11 15 |
(14) 2.6 |
380 373 370 |
(374) 5.1 |
21 26 28 |
(25) 3.6 |
5 6 7 |
(6) 1.0 |
|
+ |
15 |
150 125 163 |
(146) 19.3 |
17 14 12 |
(14) 2.5 |
361 352 376 |
(363) 12.1 |
24 21 25 |
(23) 2.1 |
5 7 12 |
(8) 3.6 |
|
+ |
50 |
156 158 149 |
(154) 4.7 |
9 6 12 |
(9) 3.0 |
332 362 403 |
(366) 35.6 |
22 22 19 |
(21) 1.7 |
5 6 5 |
(5) 0.6 |
|
+ |
150 |
127 140 150 |
(139) 11.5 |
13 19 17 |
(16) 3.1 |
419 369 363 |
(384) 30.7 |
33 20 18 |
(24) 8.1 |
9 3 6 |
(6) 3.0 |
|
+ |
500 |
119 136 164 |
(140) 22.7 |
17 15 18 |
(17) 1.5 |
389 337 345 |
(357) 28.0 |
19 24 20 |
(21) 2.6 |
12 3 6 |
(7) 4.6 |
|
+ |
1500 |
133 130 121 |
(128) 6.2 |
8 6 9 |
(8) 1.5 |
404 412 304 |
(373) 60.2 |
23 25 19 |
(22) 3.1 |
3 14 2 |
(6) 6.7 |
|
+ |
5000 |
98 82 77 |
(86) 11.0 |
6 6 7 |
(6) 0.6 |
75 88 94 |
(86) 9.7 |
7 25 16 |
(16) 9.0 |
0 0 0 |
(0) 0.0 |
|
Positive controls S9-Mix + |
Name Concentration (µg/plate) No. colonies per plate |
2AA |
2AA |
DAN |
BP |
2AA |
||||||
1 |
2 |
10 |
5 |
2 |
||||||||
1982 2219 1854 |
(2018) 185.2 |
378 376 401 |
(385) 13.9 |
789 787 779 |
(785) 5.3 |
220 262 259 |
(247) 23.4 |
284 336 315 |
(312) 26.2 |
|||
Table 4: Test Results: Experiment 2 – Without Metabolic Activation
Test Period |
From : 11 April 2003 |
To : 14 April 2003 |
||||||||||
With or without S9-Mix |
Test substance concentration (µg/plate) |
Number of revertants (mean number of colonies per plate) |
||||||||||
Base-pair substitution type |
Frameshift type |
|||||||||||
TA100 |
TA1535 |
TA102 |
TA98 |
TA1537 |
||||||||
- |
0 |
95 102 128 |
(108) 17.4# |
17 11 16 |
(15) 3.2 |
345 350 358 |
(351) 6.6 |
19 16 14 |
(16) 2.5 |
14 6 C |
(10) 5.7 |
|
- |
15 |
66 50 63 |
(60) 8.5 |
4 19 11 |
(11) 7.5 |
364 322 393 |
(360) 35.7 |
16 C 16 |
(16) 0.0 |
4 7 6 |
(6) 1.5 |
|
- |
50 |
63 66 55 |
(61) 5.7 |
4 11 7 |
(7) 3.5 |
391 383 362 |
(379) 15.0 |
16 18 17 |
(17) 1.0 |
5 9 1 |
(5) 4.0 |
|
- |
150 |
84 82 66 |
(77) 9.9 |
13 8 11 |
(11) 2.5 |
380 345 398 |
(374) 27.0 |
13 14 9 |
(12) 2.6 |
9 7 4 |
(7) 2.5 |
|
- |
500 |
82 92 95 |
(90) 6.8 |
8 17 14 |
(13) 4.6 |
376 326 397 |
(366) 36.5 |
15 12 15 |
(14) 1.7 |
7 6 5 |
(6) 1.0 |
|
- |
1500 |
86 103 85 |
(91) 10.1 |
11 8 6 |
(8) 2.5 |
326 390 329 |
(348) 36.1 |
11 13 22 |
(15) 5.9 |
9 6 3 |
(6) 3.0 |
|
- |
5000 |
0 S 0 S 0 S |
(0) 0.0 |
7 S 3 S 0 S |
(3) 3.5 |
22 23 25 |
(23) 1.5 |
0 S 0 S 0 S |
(0) 0.0 |
0 S 0 S XS |
(0) 0.0 |
|
Positive controls S9-Mix - |
Name Concentration (µg/plate) No. colonies per plate |
ENNG |
ENNG |
MMC |
4NQO |
9AA |
||||||
3 |
5 |
0.5 |
0.2 |
80 |
||||||||
419 381 274 |
(358) 75.2 |
305 227 222 |
(251) 46.5 |
1080 1040 C |
(1060) 28.3 |
160 170 151 |
(160) 9.5 |
1379 1196 1552 |
(1376) 178.0 |
|||
Table 5: Test Results: Experiment 2 – With Metabolic Activation
Test Period |
From : 11 April 2003 |
To : 14 April 2003 |
||||||||||
With or without S9-Mix |
Test substance concentration (µg/plate) |
Number of revertants (mean number of colonies per plate) |
||||||||||
Base-pair substitution type |
Frameshift type |
|||||||||||
TA100 |
TA1535 |
TA102 |
TA98 |
TA1537 |
||||||||
+ |
0 |
94 113 115 |
(107) 11.6# |
11 8 11 |
(10) 1.7 |
397 326 384 |
(369) 37.8 |
22 26 24 |
(24) 2.0 |
5 7 7 |
(6) 1.2 |
|
+ |
15 |
135 92 107 |
(111) 21.8 |
5 11 14 |
(10) 4.6 |
380 399 396 |
(392) 10.2 |
23 23 29 |
(25) 3.5 |
12 7 5 |
(8) 3.6 |
|
+ |
50 |
128 107 129 |
(121) 12.4 |
13 8 14 |
(12) 3.2 |
345 381 394 |
(373) 25.4 |
29 29 28 |
(29) 0.6 |
7 16 7 |
(10) 5.2 |
|
+ |
150 |
110 137 114 |
(120) 14.6 |
8 11 11 |
(10) 1.7 |
422 424 425 |
(424) 1.5 |
21 29 33 |
(28) 6.1 |
6 11 4 |
(7) 3.6 |
|
+ |
500 |
116 119 134 |
(123) 9.6 |
5 12 13 |
(10) 4.4 |
399 405 361 |
(388) 23.9 |
31 C 29 |
(30) 1.4 |
6 3 5 |
(5) 1.5 |
|
+ |
1500 |
76 85 92 |
(84) 8.0 |
11 7 9 |
(9) 2.0 |
434 380 376 |
(397) 32.4 |
36 22 25 |
(28) 7.4 |
7 5 6 |
(6) 1.0 |
|
+ |
5000 |
46 46 37 |
(43) 5.2 |
5 5 7 |
(6) 1.2 |
39 74 44 |
(52) 18.9 |
15 12 21 |
(16) 4.6 |
3 7 1 |
(4) 3.1 |
|
Positive controls S9-Mix + |
Name Concentration (µg/plate) No. colonies per plate |
2AA |
2AA |
DAN |
BP |
2AA |
||||||
1 |
2 |
10 |
5 |
2 |
||||||||
1099 1410 1350 |
(1286) 165.0 |
236 292 243 |
(257) 30.5 |
1135 1103 1090 |
(1109) 23.2 |
383 477 536 |
(465) 77.2 |
876 723 678 |
(759) 103.8 |
|||
S Sparse bacterial background lawn
V Very weak bacterial background lawn
# Standard deviation
C Contaminated
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative
Ammonium thioglycolate was considered to be non-mutagenic under the conditions of this test. - Executive summary:
Ammonium thioglycolate was tested in a bacterial gene mutations assay performed following a protocol compliant with the OECD guideline # 471. The direct plate incorporation procedure was performed with Salmonella typhimurium TA 98, TA 100, TA 102, TA 1535 and TA 1537 at concentrations up to 5,000 µg/plate. Cytotoxic effects were observed in the absence and in the presence of a metabolic activator (Aroclor 1254-induced rat liver S9) at the concentration of 5,000 µg/plate. Ammonium thioglycolate did not induce mutations in the bacterial mutation test in either the absence or presence of metabolic activator in any strain tested. The positive and negative controls included in the experiment showed the expected results.
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