Nitrosylsulphuric acid

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010 -01-12 - 2010-01-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study was performed and reported in compliance with the OECD Guideline 471, EC No. 440/2008 B13/14 and German principles of GLP. Guideline-conform study under GLP without deviations.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Principles of method if other than guideline:

Not applicable.

GLP compliance:

yes (incl. QA statement)

Remarks:

GLP certificate of the test facility is available

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Nitrosylsulphuric acid

EC Number:

231-964-2

EC Name:

Nitrosylsulphuric acid

Cas Number:

7782-78-7

Molecular formula:

HNO5S

IUPAC Name:

hydroxy(nitrosooxy)sulfane dioxide

Test material form:

other: liquid

Details on test material:

- Name of test material (as cited in study report): LZ5593
- Physical state: liquid
- Expiration date of the lot/batch: November 19, 2014
- Storage condition of test material: At room temperature

Method

Target gene:

Not applicable.

Species / strain

Species / strain / cell type:

other: TA 1535, TA 1537, TA 98, TA 100, TA 102

Additional strain / cell type characteristics:

not applicable

Metabolic activation:

with and without

Metabolic activation system:

Phenobarbital/ß-Naphthoflavone induced rat liver S9

Test concentrations with justification for top dose:

3, 10; 33; 100; 333; 1000; 2500; and 5000 µg/plate / pre-experiment/experiment I

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: deionised water
- Justification for choice of solvent/vehicle: better than others

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: sodium azide; 4-nitro-o-phenylene-diamine; methyl methane sulfonate, 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: plate incorporation


DURATION- Preincubation period: 1 hour
- Exposure duration: 72 hours


NUMBER OF REPLICATIONS: 3 plates


DETERMINATION OF CYTOTOXICITY
A reduction in the number of spontaneous revertants (below the induction factor of 0.5) or a clearing of the bacterial background lawn.

Evaluation criteria:

A test item is considered as a mutagen if a biologically relevant increase in the number of revertants exceeding the threshold of twice (strains TA 98, TA 100, and TA 102) or thrice (strains TA 1535 and TA 1537) the colony count of the corresponding solvent control is observed.
A dose dependent increase is considered biologically relevant if the threshold is exceeded at more than one concentration.
An increase exceeding the threshold at only one concentration is judged as biologically relevant if reproduced in an independent second experiment.
A dose dependent increase in the number of revertant colonies below the threshold is regarded as an indication of a mutagenic potential if reproduced in an independent second experiment. However, whenever the colony counts remain within the historical range of negative and solvent controls
such an increase is not considered biologically relevant.

Statistics:

According to the OECD guideline 471, a statistical analysis of the data is not mandatory.

Results and discussion

Test results

Species / strain:

other: TA 1535, TA 1537, TA 98, TA 100, TA 102

Metabolic activation:

with and without

Genotoxicity:

positive

Remarks:

in strain TA 1535 with and without S9 mix

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No precipitation
- Other confounding effects:
COMPARISON WITH HISTORICAL CONTROL DATA: performed
ADDITIONAL INFORMATION ON CYTOTOXICITY: no

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

 Summary of Results Pre-Experiment and Experiment I

Study Name: 1301600	Study Code: Harlan CCR 1301600
Experiment: 1301600 VV Plate	Date Plated: 12/01/2010
Assay Conditions:	Date Counted: 15/01/2010

Metabolic Activation	Test Group	Dose Level (per plate)		Revertant Colony Counts (Mean ±SD)
				TA 1535	TA 1537	TA 98	TA 100	TA 102
Without Activation	Deionised water			20 ± 2	12 ± 4	30 ± 7	146 ± 12	510 ± 16
	Untreated			17 ± 4	9 ± 1	28 ± 8	143 ± 18	476 ± 36
	LZ5593	3 µg		15 ± 5	11 ± 2	31 ± 4	133 ± 8	454 ± 20
		10 µg		20 ± 7	12 ± 4	29 ± 3	128 ± 5	486 ± 12
		33 µg		18 ± 1	10 ± 2	27 ± 5	145 ± 6	473 ± 20
		100 µg		23 ± 7	10 ± 5	34 ± 9	124 ± 2	476 ± 42
		333 µg		18 ± 3	9 ± 3	33 ± 18	141 ± 8	473 ± 25
		1000 µg		42 ± 5	6 ± 4	33 ± 4	155 ± 23	470 ± 73
		2500 µg		74 ± 6	11 ± 2	35 ± 8	192 ± 12	531 ± 42
		5000 µg		106 ± 13	11 ± 3	30 ± 12	230 ± 36	498 ± 18
	NaN3	10 µg		2332 ± 40			2485 ± 198
	4-NOPD	10 µg				298 ± 15
	4-NOPD	50 µg			98 ± 1
	MMS	3.0 µL						5941 ± 298
With Activation	Deionised water			24 ± 6	13 ± 1	39 ± 7	153 ± 11	637 ± 30
	Untreated			23 ± 3	10 ± 4	34 ± 6	164 ± 15	670 ± 18
	LZ5593	3 µg		18 ± 6	14 ± 2	43 ± 8	151 ± 9	651 ± 38
		10 µg		19 ± 3	12 ± 1	38 ± 6	161 ± 12	681 ± 23
		33 µg		21 ± 3	15 ± 1	43 ± 2	155 ± 7	674 ± 21
		100 µg		23 ± 3	16 ± 4	49 ± 6	149 ± 22	650 ± 35
		333 µg		29 ± 7	15 ± 5	43 ± 4	153 ± 9	655 ± 21
		1000 µg		40 ± 8	10 ± 1	42 ± 3	171 ± 23	687 ± 34
		2500 µg		67 ± 10	11 ± 3	43 ± 8	195 ± 6	669 ± 55
		5000 µg		91 ± 7	20 ± 6	41 ± 12	202 ± 22	613 ± 5
	2-AA	2.5 µg		633 ± 97	350 ± 68	2604 ± 42	2296 ± 88
	2-AA	10.0 µg						3129 ± 154

Key to Positive Controls
NaN3 2-AA MMS 4-NOPD	sodium azide 2-aminoanthracene methyl methane sulfonate 4-nitro-o-phenylene-diamine

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
positive in strain TA 1535 with and without S9 mix

In conclusion, it can be stated that during the described mutagenicity test and under the experimental conditions reported, the test item did induce gene mutations by base pair changes in the genome of strain TA 1535.

Executive summary:

The study was performed in January 2010 and reported in compliance with the OECD Guideline 471, EC No. 440/2008 B13/14 and German principles of GLP. Guideline-conform study under GLP without deviations.

The assay was performed with and without liver microsomal activation. Each concentration and the controls, were tested in triplicate. The test item was tested at the following concentrations:

Pre-Experiment/Experiment I:    3; 10; 33; 100; 333; 1000; 2500; and 5000 µg/plate

The plates incubated with the test item showed normal back­ground growth up to 5000 µg/plate with and without S9 mix in all strains used.

No toxic effects, evident as a reduction in the number of revertants (below the indication factor of 0.5), occurred in the test groups with and without metabol­ic activation.

A biological relevant increases in revertant colony numbers were observed following treatment with LZ5593 in strain TA 1535 in the presence and absence of metabolic activation (S9 mix). The number of colonies exceeded the threshold of thrice the number of the corresponding solvent control at concentrations at 2500 µg/plate and at 5000 µg/plate.