Disodium tartrate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Ames test (OECD 471): negative in S. typhimurium TA 1535, TA 1537, TA 1538, TA 98 and TA 100 and E. coli WP2 uvrA with and without metabolic activation

RA from source substance potassium sodium tartarate (CAS 304 -59 -6)

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study with acceptable restrictions

Remarks:

/ 2-AA used as sole indicator of S9 mix efficacy; duplicate plating without scientific justification; only 1 plate for some positive controls; no result of 2-AA (+S9 mix) in 1 exp (TA 1537, TA 1538 and WP2 uvr A); AF-2 used as positive control of WP2 uvr A

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Version / remarks:

21 July 1997

Principles of method if other than guideline:

- Principle of test: similar to OECD 471 but with several differences:
2-AA was used as sole indicator of S9 mix efficacy; duplicate plating without scientific justification; only 1 plate for some positive controls; no result of 2-AA (+S9 mix) in 1 exp (TA 1537, TA 1538 and WP2 uvr A); AF-2 used as positive control of WP2 uvr A

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Target gene:

his operon, trp operon

Species / strain / cell type:

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

Species / strain / cell type:

S. typhimurium TA 1538

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of male rats, treated with Aroclor 1254 (500 mg/kg bw)

Test concentrations with justification for top dose:

Based on a range-finding study (performed in tester strain TA 100; doses applied: 0.3 - 10000 μg/plate), the following concentrations were used in the main experiments:
TA 1535 / TA 100:
1st exp.: 33.3, 100, 333.3, 1000, 3333.3 and 10000 µg/plate with and without metabolic activation
2nd exp.: 0.3, 3.3, 33.3, 100, 333.3, 1000, 3333.3 and 10000 µg/plate with and without metabolic activation

TA 1537 / TA 1538 / TA 98 / WP2 uvr A:
1st exp.: 0.3, 3.3, 33.3, 100, 333.3, 1000, 3333.3 and 10000 µg/plate with and without metabolic activation
2nd exp.: 33.3, 100, 333.3, 1000, 3333.3 and 10000 µg/plate with and without metabolic activation

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: potassium phosphate buffer

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Remarks:

potassium phosphate buffer (0.067 M, pH 7)

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

other: AF-2 (2-(2-Furyl)-3-(5-nitro-2-furyl) acrylamide), 2AA (2-Aminoanthracene)

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation) (Range Finding Test, first and second experiment)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 2

DETERMINATION OF CYTOTOXICITY
- Method: not provided

Statistics:

Mean values were calculated.

Key result

Species / strain:

S. typhimurium TA 98

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 applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 100

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 applicable

Positive controls validity:

valid

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 applicable

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 applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1538

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 applicable

Positive controls validity:

valid

Key result

Species / strain:

E. coli WP2 uvr A

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 applicable

Positive controls validity:

valid

Additional information on results:

RANGE-FINDING/SCREENING STUDIES: A range finding assay was performed in TA 100. No mutagenicity and toxicity were observed.

Table 1: Summary of test results (Exp. 1, Plate Incorporation Method)

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 2 plates)
		Frameshift type			Base-pair substitution type
		TA98	TA1537	TA1538	TA100	TA1535	WP2 uvrA
–	Solvent control (potassium phosphate buffer)	24*	16*	13*	204*	41*	50*
	0.3	22	13	15	-	-	49
	3.3	23	15	13	-	-	51
	33.3	21	18	19	204	49	47
	100	24	12	10	198	43	41
	333.3	22	14	25	201	38	45
	1000	22	11	11	174	48	36
	3333.3	28	19	14	205	47	48
	10000	14	14	16	183	37	53
	Positive controls (µg/plate)	2-NF (5)	9AA (50)	2-NF (5)	SA (0.5)	SA (0.5)	AF2 (0.1)
	Mean No. of colonies/plate (average of 2 plates)	359	185	692	411**	264**	103
	Positive controls (µg/plate)	2AA (2.5)	2AA (1)	2AA (1)	2AA (2.5)	2AA (1)	2AA (10)
	Mean No. of colonies/plate (average of 2 plates)	39**	nt	nt	181**	37**	72**
+	Solvent control (potassium phosphate buffer)	36*	11*	22*	252*	41*	102*
	0.3	47	14	21	-	-	47
	3.3	34	18	17	-	-	53
	33.3	40	16	32	244	69	48
	100	31	18	32	219	78	51
	333.3	38	14	27	246	67	52
	1000	45	12	25	259	59	59
	3333.3	34	13	24	256	62	49
	10000	35	19	34	239	52	54
	Positive controls (µg/plate)	2AA (2.5)	2AA (1)	2AA (1)	2AA (2.5)	2AA (1)	2AA (10)
	Mean No. of colonies/plate (average of 2 plates)	965**	nt	nt	360**	119**	166**

2AA = 2-aminoanthracene

2-NF = 2-nitrofluorene

9AA = 9-aminoacridine

AF-2 = 2-(2-Furyl)-3-(5-nitro-2-furyl) acrylamide

SA = sodium azide

* = 4 results

** = 1 result

Table 2: Summary of test results (Exp. 2, Plate Incorporation Method)

With or without S9-Mix	Test substance concentration (μg/plate)	Mean number of revertant colonies per plate (average of 2 plates)
		Frameshift type			Base-pair substitution type
		TA98	TA1537	TA1538	TA100	TA1535	WP2 uvrA
–	Solvent control (potassium phosphate buffer)	15*	5*	11*	77*	15*	33*
	0.3	-	-	-	77	14	-
	3.3	-	-	-	76	17	-
	33.3	21	5	14	78**	14	35
	100	20	4	9	70	11	36
	333.3	16	5	18	77	14	41
	1000	21	5	11	70	11	34
	3333.3	17	3	15	82	16	36
	10000	15	5	17	67	12	39
	Positive controls (µg/plate)	2-NF (5)	9AA (50)	2-NF (5)	SA (0.5)	SA (0.5)	AF2 (0.1)
	Mean No. of colonies/plate (average of 2 plates)	359**	154**	591**	417	300	1581
	Positive controls (µg/plate)	2AA (2.5)	2AA (1)	2AA (1)	2AA (2.5)	2AA (1)	2AA (10)
	Mean No. of colonies/plate (average of 2 plates)	18**	3**	19**	89**	7**	nt
+	Solvent control (potassium phosphate buffer)	27*	7*	21*	85*	5*	40*
	0.3	-	-	-	65	9	-
	3.3	-	-	-	84	7	-
	33.3	44	9	26	75	7	44
	100	36	9	25	66	6	58
	333.3	27	8	25	84	6	43
	1000	28	8	24	71	5	52
	3333.3	29	5	25	72	6	49
	10000	27	6	21	65	7	57
	Positive controls (µg/plate)	2AA (2.5)	2AA (1)	2AA (1)	2AA (2.5)	2AA (1)	2AA (10)
	Mean No. of colonies/plate (average of 2 plates)	104**	41**	89**	882**	76**	nt

2AA = 2-aminoanthracene

2-NF = 2-nitrofluorene

9AA = 9-aminoacridine

AF-2 = 2-(2-Furyl)-3-(5-nitro-2-furyl) acrylamide

SA = sodium azide

* = 4 results

** = 1 result

Reference 2

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Justification for type of information:

refer to analogue justification provided in IUCLID section 13

Reason / purpose for cross-reference:

read-across source

Key result

Species / strain:

S. typhimurium TA 98

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 applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 100

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 applicable

Positive controls validity:

valid

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 applicable

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 applicable

Positive controls validity:

valid

Key result

Species / strain:

S. typhimurium TA 1538

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 applicable

Positive controls validity:

valid

Key result

Species / strain:

E. coli WP2 uvr A

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 applicable

Positive controls validity:

valid

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Genetic toxicity

 

Justification for read-across

There are no data for genetic toxicity in bacteria are available for  disodium tartrate (CAS 868 -18 -8). To fulfil the standard data requirements defined in Regulation (EC) No. 1907/2006, Annex VII, 8.4, read-across from an appropriate substance is conducted in accordance with Regulation (EC) No 1907/2006, Annex XI.

According to Article 13 (1) of Regulation (EC) No 1907/2006, "information on intrinsic properties of substances may be generated by means other than tests, provided that the conditions set out in Annex XI are met”. In particular for human toxicity, information shall be generated whenever possible by means other than vertebrate animal tests, which includes the use of information from structurally related substances (grouping or read-across) “to avoid the need to test every substance for every endpoint”.

 

For each specific endpoint the source substance(s) structurally closest to the target substance is/are chosen for read across, with regard to the requirements of adequacy and reliability of the available data. Structural similarities and similarities in properties and/or activities of the source and target substances are the basis of read-across. A detailed justification for the analogue read- across approach is provided in the technical dossier (see IUCLID Section 13).

 

As no reliable measured/experimental data are available on genetic toxicity in bacteria with  disodium tartrate (CAS 868 -18 -8), read-across to reliable data on the analogue substance potassium sodium tartrate (CAS 304 -59 -6) was conducted.

 

A bacterial gene mutation assay with the test substance was performed, similar to OECD Guideline 471 (Simmon and Eckford, 1979). In this study the substance was not mutagenic in any of the six strains (TA 1535, TA 1537, TA 1538, TA 98, TA 100 and WP2 uvrA) tested with and without metabolic activation.

Justification for classification or non-classification

Based on the analogue read-across approach and on the available data, there is no indication that the substance induces genetic toxicity in bacteria. Nevertheless, no final decision on classification for genetic toxicity according to Regulation (EC) 1272/2008 can be made, as no information on chromosomal aberration and mutagenicity in mammalian cells/in vivo is available.