1,4-dibromobutane

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:

guideline study

Justification for type of information:

Data is from experimental report.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1987

Report date:

1987

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Principles of method if other than guideline:

To evaluate the mutagenic potential of test chemical in Salmonella typhimurium strain TA 1535, TA 100, TA 1537, TA 98 by Ames test.

GLP compliance:

not specified

Type of assay:

bacterial forward mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

1,4-dibromobutane

EC Number:

203-775-5

EC Name:

1,4-dibromobutane

Cas Number:

110-52-1

Molecular formula:

C4H8Br2

IUPAC Name:

1,4-dibromobutane

Test material form:

liquid

Details on test material:

IUPAC name:1,4-dibromobutane
Mol. formula: C4H8Br2
Molecular Weight: 215.915
Smiles: C(CCBr)CBr
InChI: 1S/C4H8Br2/c5-3-1-2-4-6/h1-4H2

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA 1535, TA 100, TA 1537, TA 98

Details on mammalian cell type (if applicable):

Not applicable.

Additional strain / cell type characteristics:

other: All strains show a considerably reduced hydrophilic polysaccharide layer (rfa), which leads to an increase in permeability to lipophilic substances.

Cytokinesis block (if used):

not specified

Metabolic activation:

with and without

Metabolic activation system:

S-9 mix was prepared from Sprague-Dawley rats livers induced with Aroclor 1254.

Test concentrations with justification for top dose:

1st Experiment
Strains: TA 1535. TA 100. TA 1537, TA 98
Dose.: 0, 20, 100, 500. 2500 and 5000 µg/plate

2nd Experiment
Strains: TA 100, TA 98
Domes: 0, 20. 100, 500 and 1000 µg/plate

3rd Experiment
Strain: TA 100
Doses: 0, 50, 100, 500, 1000 and 2500 µg/plate

4th Experiment
Strains: TA 1535, JA 1537
Doses: 0, 20, 100, 500, 1000 and 1500 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Complete solubility of the test substance in DMSO.

Controls

Untreated negative controls:

yes

Remarks:

DMSO

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: + S-9 mix 10 µg 2-aminoanthracene for the strains TA 100, TA 98, TA 1537 and TA 1535 - S-9 mix 5µg N-methyl-N-nitro-Nnitroso;TA 100 and TA 1535 10 µg 4 -nitro-o-phenylenediamine ;strain TA 98 100 µg 9-aminoacridine chloride monohydrate ; TA 1537.

Details on test system and experimental conditions:

METHOD OF APPLICATION: Standard Plate method
- Cell density at seeding (if applicable): >108 bacteria/ml

DURATION
- Preincubation period: 16 hours
- Exposure duration:2days

DETERMINATION OF CYTOTOXICITY
- Method: A bacteriotoxic effect OTHER EXAMINATIONS:

Evaluation criteria:

A substance to be characterized as positive in the Ames test has to fulfill the following requirements:
- doubling of the spontaneous mutation rate
- dose-response relationship
- reproducibility of the results.

Statistics:

Yes ,,Mean ±standard deviation was observed.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium, other: TA 1535, TA 100, TA 1537 and TA 98.

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Positive controls validity:

other: As though the test substace was used as a positive control

Additional information on results:

No mutagenic effect were observed in the salmonella typhimurium strain TA 100 used in expriment number three , when exposed to test substance.

Remarks on result:

other: No mutagenic effect were observed.

Any other information on results incl. tables

TABLE 1

STRAIN: TA 1535

AMES TEST WITH: 86/221

METHOD: STANDARD PLATE TEST

 

DOSE MCG/PL	REVERTANTS/PLATES						TITER DIL	QUOTIENT
	-S9	M	SD	+S9*	M	SD	EXP-6	-S9	+S9*
NEGATIVE CONTROL DMSO	15 18 21	18	3	21 17 19	19	2	45 41 3	1.0	1.0
20	18 16 14	16	2	21 21 26	23	3		0.9	1.2
100	18 20 14	17	3	22 22 24	23	1		1.0	1.2
500	20 24 21	22	2	28 28 24	27	2		1.2	1.4
2500	B B B	-	-	13B 12B 8B	11	3	0 0 0	-	0.6
5000	B B B	-	-	B B B	-	-	0 0 0	-	-
POSITIVE CONTROL 2-AA 10				320 265 242	276	40			14.5
POSITIVE CONTROL MNNG 5	1960 2350 2300	2203	212					122.4

* : S-9 FRACTION/COFACTORS = 3: 7         EXP :EXP. TO 10

B: REDUCED his BACKGROUND           

           

 

 

 

 

 

 

 

TABLE 2

STRAIN: TA 100

AMES TEST WITH: 86/221

METHOD: STANDARD PLATE TEST

DOSE MCG/PL	REVERTANTS/PLATES						TITER DIL	QUOTIENT
	-S9	M	SD	+S9*	M	SD	EXP-6	-S9	+S9*
NEGATIVE CONTROL DMSO	94 109 117	107	12	110 104 107	107	3	15 28 11	1.0	1.0
20	113 119 111	114	4	102 118 125	115	12		1.1	1.1
100	116 117 111	112	7	119 125 113	119	6		1.1	1.1
500	113 124 146	128	17	137 139 138	138	1		1.2	1.3
2500	B B B	-	-	74B 73B 80B	76	4	0 0 0	-	0.7
5000	B B B	-	-	B B B	-	-	0 0 0	-	-
POSITIVE CONTROL 2-AA 10				1780 1860 1910	1850	66			17.3
POSITIVE CONTROL MNNG 5	1740 1930 1850	1840	95					17.3

 

* : S-9 FRACTION/COFACTORS = 3: 7         EXP :EXP. TO 10

B: REDUCED his BACKGROUND                       

 

 

 

 

 

 

TABLE 3

STRAIN: TA 1537

AMES TEST WITH: 86/221

METHOD: STANDARD PLATE TEST

DOSE MCG/PL	REVERTANTS/PLATES						TITER DIL	QUOTIENT
	-S9	M	SD	+S9*	M	SD	EXP-6	-S9	+S9*
NEGATIVE CONTROL DMSO	13 13 10	12	2	8 10 10	9	1	16 13 11	1.0	1.0
20	9 13 8	10	3	15 11 8	11	4		0.8	1.2
100	11 10 11	11	1	8 10 8	9	1		0.9	0.9
500	16 9 11	12	4	9 9 16	11	4		1.0	1.2
2500	B B B	-	-	5B 8B 9B	7	2	0 0 0	-	0.8
5000	B B B	-	-	B B B	-	-	0 0 0	-	-
POSITIVE CONTROL 2-AA 10				199 194 195	196	3			21.0
POSITIVE CONTROL MNNG 5	744 661 833	746	86					62.2

 

* : S-9 FRACTION/COFACTORS = 3: 7         EXP :EXP. TO 10

B: REDUCED his BACKGROUND                       

 

 

 

 

 

 

TABLE 4

STRAIN: TA 98

AMES TEST WITH: 86/221

METHOD: STANDARD PLATE TEST

DOSE MCG/PL	REVERTANTS/PLATES						TITER DIL	QUOTIENT
	-S9	M	SD	+S9*	M	SD	EXP-6	-S9	+S9*
NEGATIVE CONTROL DMSO	22 21 21	21	1	39 32 31	34	4	62 53 57	1.0	1.0
20	21 29 22	24	4	31 34 32	32	2		1.1	1.0
100	21 21 -	21	0	40 32 29	34	6		1.0	1.0
500	20 24 23	22	2	35 30 28	31	4		1.0	0.9
2500	B B B	-	-	17B 23B 17B	19	3	0 0 0	-	0.6
5000	B B B	-	-	B B B	-	-	0 0 0	-	-
POSITIVE CONTROL 2-AA 10				1992 1800 2000	1907	101			56.1
POSITIVE CONTROL MNNG 5	781 839 899	840	59					39.4

 

* : S-9 FRACTION/COFACTORS = 3: 7         EXP :EXP. TO 10

B: REDUCED his BACKGROUND                       

 

Table – 5

AMES TEST WITH 86/221

METHOD: STANDARD PLATE TEST STRAIN: TA 100

DOSE MCG/ PL	REVERTANTS / PLATE						QUOT IENT
	-S9	M	SD	+S9*	M	SD	-S9	+S9*
NEGATIVE CONTROL DMSO	116	114	10	105	103	8	1.0	1.0
	103			94
	123			109
20	99	102	13	115	110	8	0.9	1.1
	91			101
	117			113
100	93	103	10	97	104	8	0.9	1.0
	112			104
	103			112
500	109	111	10	112	106	6	1.0	1.0
	102			103
	121			102
1000	93	98	9	141	129	10	0.9	1.3
	92			123
	108			124
POSITIVE CONTROL 2-AA				2100	2033	76		19.8
				2050
				1950
10
POSITIVE CONTROL MNNG	1670	1703	42				14.9
	1750
	1690
5

* : S9 FRACTION/COFACTORS = 3. 7

 

Table – 6

AMES TEST WITH 86/33

METHOD: STANDARD PLATE TEST STRAIN: TA 100

DOSE MCG/ PL	REVERTANTS / PLATE						QUOT IENT
	-S9	M	SD	+S9*	M	SD	-S9	+S9*
NEGATIVE CONTROL DMSO	116	114	10	105	103	8	1.0	1.0
	103			94
	123			109
20	122	116	6	117	113	6	1.0	1.1
	113			-
	112			108
100	96	110	21	125	132	8	1.0	1.3
	100			140
	134			132
500	127	128	17	137	152	17	1.1	1.5
	112			148
	145			171
1000	149	145	7	207	206	2	1.3	2.0
	150			206
	137			204
POSITIVE CONTROL 2-AA				2100	2033	76		19.8
				2050
				1950
10
POSITIVE CONTROL MNNG	1670	1703	42				14.9
	1750
	1690
5

* : S9 FRACTION/COFACTORS = 3. 7 - : CONTAMINATION

Table – 7

AMES TEST WITH 86/221

METHOD: STANDARD PLATE TEST STRAIN: TA 98

DOSE MCG/ PL	REVERTANTS / PLATE						QUOT IENT
	-S9	M	SD	+S9*	M	SD	-S9	+S9*
NEGATIVE CONTROL DMSO	25	23	2	35	34	4	1.0	1.0
	22			38
	21			30
20	30	25	6	34	32	2	1.1	0.9
	25			32
	19			31
100	24	20	4	40	38	3	0.7	1.1
	19			36
	17			-
500	17	18	2	29	29	2	0.8	0.8
	20			30
	17			27
1000	12	10	3	33	30	3	0.5	0.9
	7			30
	12			27
POSITIVE CONTROL 2-AA				1433	1420	26		41.4
				1440
				1390
10
POSITIVE CONTROL MNNG	1060	1150	95				50.7
	1140
	1250
10

* : S9 FRACTION/COFACTORS = 3. 7 - : CONTAMINATION

Table – 7

AMES TEST WITH 86/221

METHOD: STANDARD PLATE TEST STRAIN: TA 100

DOSE MCG/ PL	REVERTANTS / PLATE						TITER DIL	QUOT IENT
	-S9	M	SD	+S9*	M	SD	EXP-6	-S9	+S9*
NEGATIVE CONTROL DMSO	117	119	8	110	108	3	14	1.0	1.0
	128			104			15
	112			109			18
50	91	105	16	121	113	7		0.9	1.0
	122			109
	101			108
100	123	131	7	103	124	26		1.1	1.1
	132			115
	137			153
500	113	126	14	108	129	18		1.1	1.2
	124			140
	140			138
1000	127	122	8	139	154	13	22	1.0	1.4
	113			162			31
	127			162			17
5000	B	-	-	77	74	9	21	-	0.7
	B			81			16
	B			63			11
POSITIVE CONTROL 2-AA				1660	1630	79			15.1
				1690
				1540
10
POSITIVE CONTROL MNNG	2150	2015	191					16.9
	1880
	-
5

 

*: S59 FRACTION/COFACTORS = 3.7 EXP : EXP. T0 10

-: CONTAMINATION B: REDUCED his- BACKOROUND

Table – 9

AMES TEST WITH 87/33

METHOD: STANDARD PLATE TEST STRAIN: TA 100

DOSE MCG/ PL		REVERTANTS / PLATE						QUOT IENT
	-S9	M	SD	+S9*	M	SD	TITER DIL Exp-6	-S9	+S9*
NEGATIVE CONTROL DMSO	117	119	8	110	108	3	14	1.0	1.0
	128			104			15
	112			109			18
50	127	119	7	120	119	8		1.0
	115			126
	114			110
100	116	113	3	136	132	5		0.9	1.2
	110			127
	113			133
500	135	141	9	132	156	26		1.2	1.4
	152			152
	137			183
1000	182	173	18	157	170	19	33	1.5	1.6
	153			192			31
	137			161			34
2500	211	210	16	235	250	13	26	1.8	2.3
	225			253			32
	193			261			31
POSITIVE CONTROL 2-AA				1660	1630	79			15.1
				1690
				1540
10
POSITIVE CONTROL MNNG	2150	2015	191					16.9
	1880
	-
5

 

 

 

 

 

 

 

 

Table – 10

AMES TEST WITH 86/221

METHOD: STANDARD PLATE TEST STRAIN: TA1535

 

DOSE MCG/ PL		REVERTANTS / PLATE						QUOT IENT
	-S9	M	SD	+S9*	M	SD	TITER DIL Exp-6	-S9	+S9*
NEGATIVE CONTROL DMSO	20	19	3	23	20	3	49	1.0	1.0
	16			17			44
	21			19			51
20	21	23	2	17	17	2		1.2	0.8
	24			15
	23			18
100	20	20	2	19	17	2		1.1	0.8
	22			16
	18			15
500	18	19	3	19	20	3		1.0	1.0
	17			18
	23			24
1000	22	20	3	18	18	1	55	1.1	0.9
	17			17			64
	21			19			48	1.2	1.0
1500	24	23	1	20	20	4	56
	22			24			57
	22			17			48
POSITIVE CONTROL 2-AA				577	481	140			24.4
				545
				320
10
POSITIVE CONTROL MNNG	2050	2233	176					117.5
	2250
	2400
5

 

 

 

 

 

 

 

 

 

Table – 11

AMES TEST WITH 86/221

METHOD: STANDARD PLATE TEST STRAIN: TA1535

 

DOSE MCG/ PL		REVERTANTS / PLATE						QUOT IENT
	-S9	M	SD	+S9*	M	SD	TITER DIL Exp-6	-S9	+S9*
NEGATIVE CONTROL DMSO	14	14	2	14	15	1	33	1.0	1.0
	12			15			27
	16			15			27
20	15	15	1	14	14	1		1.1	1.0
	14			13
	16			15
100	14	13	1	13	13	3		1.0	0.9
	13			16
	13			10
500	15	13	2	10	13	3		0.9	0.9
	13			15
	11			13
1000	10	11	2	12	13	2	32	0.8	0.9
	11			12			33
	13			16			44
1500	12	14	2	14	15	2	24	1.0	1.0
	14			18			32
	15			14			30
POSITIVE CONTROL 2-AA				203	221	16			15.1
				234
				226
10
POSITIVE CONTROL MNNG	661	702	109					50.1
	825
	619
5

 

 

Applicant's summary and conclusion

Conclusions:

Test chemical was evaluated for its mutagenic potential in Salmonella typhimurium strain TA 1535, TA 100, TA 1537, TA 98 by Ames test. Test chemical, which is known to be positive in the Ames test, was investigated in parallel as a positive control substance (chemical class control) using the strain TA 100. The test result was considered to be negative in the presence and absence of metabolic activation.

Executive summary:

Test chemical was evaluated for its mutagenic potential in Salmonella typhimurium strain TA 1535, TA 100, TA 1537, TA 98 by Ames test. The test chemical, which is known to be positive in the Ames test, was investigated in parallel as a positive control substance (chemical class control) using the strain TA 100. The test result was considered to be negative in the presence and absence of metabolic activation.Genetic toxicity study was assessed for its possible mutagenic potential. For this purpose AMES test was performed according to guideline OECD 471. The test material was exposed at different concentration in 4 individual experiments. These concentrations are mentioned below :

1st Experiment

Strains: TA 1535. TA 100. TA 1537, TA 98

Doses: 0, 20, 100, 500. 2500 and 5000 µg/plate

 

2nd Experiment

Strains: TA 100, TA 98

Doses: 0, 20. 100, 500 and 1000 µg/plate

 

3rd Experiment

Strain: TA 100

Doses: 0, 50, 100, 500, 1000 and 2500 µg/plate

 

4th Experiment

Strains: TA 1535, JA 1537

Doses: 0, 20, 100, 500, 1000 and 1500 µg/plate

The test material which is known to be positive in the Ames test, was investigated in parallel as a positive control substance (chemical class control) using the strain TA 100. The test material was exposed in the presence and absence of metabolic activation. No mutagenic effects were observed in any of the strain. Thereforetest chemical was considered to be non mutagenic in Salmonella typhimurium strain TA 1535, TA 100, TA 1537, TA 98 by Ames test. Hence the substance cannot be classified as gene mutant in vitro.