Sodium pyruvate

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Study period:

07 September 2017 - 06 December 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Report date:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

1997

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Version / remarks:

2008

Deviations:

no

Test guideline 3

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

Version / remarks:

df. Sofuni, 1993

Deviations:

no

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Sodium pyruvate

EC Number:

204-024-4

EC Name:

Sodium pyruvate

Cas Number:

113-24-6

Molecular formula:

C3H4O3.Na

IUPAC Name:

sodium 2-oxopropanoate

Test material form:

solid

Method

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2

Metabolic activation:

with and without

Metabolic activation system:

beta-naphthoflavone/phenobarbital induced rat liver S9 mix

Test concentrations with justification for top dose:

test concentrations: 5, 15.8, 50, 158, 500, 1580 and 5000 µg/plate
top dose: maximum recommended concentration according to current regulatory guideline (OECD 471, 1997)

Vehicle / solvent:

- Vehicle/solvent used: ultrapure 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

sodium azide

other: daunomycin: 1 µg/plate, 2-aminoanthracene: 2-10 µg/plate

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 2 days

NUMBER OF REPLICATIONS: 3 replicates for test item concentrations and positive controls, 6 replicates for solvent controls

DETERMINATION OF CYTOTOXICITY
- Method: counting numbers of revertants

OTHER:
-S9 concentration: 1st series 10%, 2nd series 30%

Evaluation criteria:

A test material was to be defined as positive or mutagenic in this assay if
- the assay is considered valid and
- a biologically relevant increase in the mean number of revertants above a threshold of 2-fold (TA 98, TA 100, WP2 uvrA) or 3-fold (TA 1535, TA 1537) as compared to the concurrent negative controls is observed
- an increase exceeding the threshold at only one concentration is considered as biologically meaningful if reproduced in a second independent experiment
- a concentration-dependent increase is considered biologically meaningful if the threshold is exceeded at more than one concentration
A test material is defined as negative or non-mutagenic in this assay if
- the assay is considered valid and
- none of the above mentioned criteria are met

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No precipitation occurred.

Any other information on results incl. tables

Table 1: Summary 1st series

Metabolic Activation	Test Material	Concentr. [µg/plate]	Revertants per plate (Mean ± SD)
			TA 98	TA 100	TA 1535	TA 1537	WP2 uvrA
Without Activation	H2O		31 ± 8	113 ± 8	24 ± 5	10 ± 3	39 ± 16
	Test item	5.00	30 ± 10	110 ± 3	24 ± 6	9 ± 2	35 ± 5
		15.8	35 ± 9	106 ± 17	30 ± 10	7 ± 1	33 ± 5
		50.0	31 ± 5	128 ± 16	28 ± 3	8 ± 2	34 ± 5
		158	38 ± 4	125 ± 17	23 ± 1	12 ± 3	32 ± 7
		500	26 ± 1	118 ± 12	24 ± 1	10 ± 1	37 ± 3
		1580	31 ± 5	125 ± 30	26 ± 5	6 ± 1	33 ± 4
		5000	30 ± 2	115 ± 8	24 ± 9	7 ± 1	36 ± 1
	DAUN	1.00	405 ± 22
	NaN3	2.00		1830 ± 110	993 ± 52
	9-AA	50.0				1376 ± 505
	NQO	2.00					1491 ± 231
With Activation	H2O		41 ± 9	117 ± 17	22 ± 4	11 ± 4	41 ± 5
	Test item	5.00	33 ± 2	106 ± 10	17 ± 4	10 ± 5	36 ± 7
		15.8	41 ± 4	118 ± 15	12 ± 3	8 ± 2	41 ± 4
		50.0	33 ± 5	119 ± 31	20 ± 8	10 ± 3	41 ± 7
		158	32 ± 5	107 ± 24	16 ± 3	12 ± 5	43 ± 9
		500	35 ± 7	109 ± 6	24 ± 3	7 ± 7	32 ± 8
		1580	29 ± 6	110 ± 14	18 ± 6	11 ± 2	35 ± 5
		5000	28 ± 3	114 ± 7	18 ± 3	7 ± 2	39 ± 5
	2-AA	2.00	827 ± 59	1511 ± 150
	2-AA	5.00			154 ± 12	397 ± 28
	2-AA	10.0					489 ± 9

Key to Positive Controls: NaN3: Sodium azide, 2-AA: 2-Aminoanthracene, 9-AA: 9-Aminoacridine, DAUN: Daunomycin, NQO: 4-Nitroquinoline-N-oxide

 

Table 2: Summary 2nd series

Metabolic Activation	Test Material	Concentr. [µg/plate]	Revertants per plate (Mean ± SD)
			TA 98	TA 100	TA 1535	TA 1537	WP2 uvrA
Without Activation	H2O		36 ± 5	102 ± 16	18 ± 3	6 ± 3	34 ± 6
	Test item	50.0	37 ± 10	94 ± 6	22 ± 6	9 ± 1	29 ± 3
		158	31 ± 8	108 ± 10	25 ± 5	5 ± 2	33 ± 7
		500	24 ± 6	100 ± 15	25 ± 4	7 ± 1	39 ± 12
		1580	27 ± 5	101 ± 4	20 ± 1	5 ± 2	31 ± 2
		5000	35 ± 3	104 ± 17	15 ± 4	8 ± 3	30 ± 4
	DAUN	1.00	199 ± 9
	NaN3	2.00		1592 ± 80	914 ± 42
	9-AA	50.0				820 ± 212
	NQO	2.00					2085 ± 77
With Activation	H2O		42 ± 8	98 ± 12	18 ± 8	11 ± 3	41 ± 10
	Test item	50.0	39 ± 13	102 ± 9	18 ± 5	10 ± 3	32 ± 14
		158	39 ± 8	96 ± 6	16 ± 2	10 ± 4	38 ± 5
		500	36 ± 1	93 ± 26	16 ± 5	15 ± 7	44 ± 1
		1580	36 ± 2	95 ± 3	12 ± 4	7 ± 3	34 ± 4
		5000	36 ± 3	94 ± 7	14 ± 3	9 ± 4	43 ± 12
	2-AA	2.00	439 ± 96	784 ± 255
	2-AA	5.00			132 ± 18	138 ± 81
	2-AA	10.0					232 ± 48

Key to Positive Controls: NaN3: Sodium azide, 2-AA: 2-Aminoanthracene, 9-AA: 9-Aminoacridine, DAUN: Daunomycin, NQO: 4-Nitroquinoline-N-oxide

 

 

Applicant's summary and conclusion

Conclusions:

The test item is considered non-mutagenic under the test conditions described.

Executive summary:

The present study was conducted to investigate the test material for its mutagenic potential in a bacterial reverse gene mutation assay in the absence and presence of a rat liver metabolizing system (S9 mix). The investigations were performed using Salmonella typhimurium tester strains TA 98, TA 100, TA 1535 and TA 1537 and Escherichia coli WP2 uvrA. The plate incorporation test with and without addition of liver S9 mix from rats pretreated with beta-Naphthoflavone/Phenobarbital was used. In this study, two experimental series were performed. The S9 mix used contained 10% S9 in the 1st and 20% S9 in the 2nd series, respectively. Solvent and positive control treatments were included for all strains. The mean numbers of revertant colonies were all within acceptable ranges for solvent control treatments, or were clearly elevated by positive control treatments, thus, showing the expected reversion properties of all strains and good metabolic activity of the S9 mix used. Following treatment of all bacteria tester strains with the test item in the absence and presence of S9 mix, no relevant increases in revertant numbers were observed. It was concluded that with and without addition of S9 mix as the exogenous metabolizing system, the test item was not mutagenic under the experimental conditions described.