Potassium hydrogen tartrate

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:

1979

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Well performed, well described study, commissioned in 1979 by USFDA and performed by reputed laboratory. Raw data available and method essentially as described in EU test method B13/14. Lack of evidence of GLP is only reason for not asigning Klimisch 1.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1979

Report date:

1979

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

yes

Remarks:

Duplicate instead of triplicate plates. However tests were performed twice.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Potassium sodium tartrate

EC Number:

206-156-8

EC Name:

Potassium sodium tartrate

Cas Number:

304-59-6

Molecular formula:

C4H6O6.K.Na

IUPAC Name:

potassium sodium tartrate

Details on test material:

F76-019 - Potassium sodium tartrate. Mallinckrodt Lot #WBPS
Purity not stated but assumed analytical grade (i.e. ca. 99%)

Method

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Arochlor-1254 stimulated rat-liver homogenate (S9 mix) according to Ames et al.

Test concentrations with justification for top dose:

A range-finding assay was performes on strain TA 100 at concentrations of 0.3 - 10.000 µg/plate.
Sunsequently two assays were performed using the following test concentrations: 0.3, 3.3, 33.3, 100.0, 333.3, 1000.0, 3333.3 and 10000 µg/plate. In the second assay only the 6 highest concentrations were used.

Vehicle / solvent:

0.067 Potassium Phosphate buffer is used as vehicle / solvent (ph 7.0).

Controls

Controls 1

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

sodium azide

Controls 2

Untreated negative controls:

yes

Positive controls:

yes

Positive control substance:

other: 2-anthramine

Controls 3

Untreated negative controls:

yes

Positive controls:

yes

Positive control substance:

9-aminoacridine

Controls 4

Untreated negative controls:

yes

Positive controls:

yes

Positive control substance:

2-nitrofluorene

Controls 5

Untreated negative controls:

yes

Positive controls:

yes

Positive control substance:

2-acetylaminofluorene

Details on test system and experimental conditions:

Test performed according to test developed by B. Ames. Briefly, innoculum from stck cultures is grown overnight a 37°C in Oxoid nutrient broth. After stationary overnight growth cultures are shaken for 3 - 4 h for optimal exponential growth.The standard plata-incorporation method is employed and test substance and controls are plated, with and without metabolic activation. Plates are incubated for 48 h at 37 °C and revertants are counted and recorded. Metabolic activation mixture contains 10% S9 fraction.

Evaluation criteria:

Not recordad, as no positive response was found. Presumably the modified two-fold rule is employed as stated in source publications and reference test method.

Results and discussion

Test results

Species / strain:

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

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Report states that in the second experiment performed abnormally high numver of revertabts were seen with strains TA 1535 and TA100 (in samples and negative controls). This was subsequently attributed to the presence of traces of ethylene oxide in the plates used.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

TABLE 1. SALMONELLA TYPHIMURIUM STRAIN TA1535. FDA COMPOUND F76-019 (Potassium Sodium Tartrate)
Compound  Metabolic  Micrograms of compound  Histidine revertants per plate     Activation added per plate   Experiment 1*     Experiment 2*           14 March 1978     7 July 1978             Average     Average Negative control −   31 36 41 19 14 15   −   37 60   14 14     +   48 35 41 4 4 5   +   37 45   7 4   Positive controls                 Sodium azide − 0,5 264     344 256 300 2-Anthramine − 1 37     7       + 1 119     76     F76-019 − 0,3       14 13 14   − 3,3       15 18 17   − 33,3 30 67 49 13 14 14   − 100 44 41 43 13 9 11   − 333,3 37 39 38 13 14 14   − 1000 51 45 48 14 7 11   − 3333,3 42 52 47 15 17 16   − 10000 24 50 37 16 8 12   + 0,3       9 9 9   + 3,3       8 6 7   + 33,3 54 84 69 4 9 7   + 100 43 113 78 5 6 6   + 333,3 63 70 67 7 4 6   + 1000 54 63 59 4 5 5   + 3333,3 56 67 62 4 8 6   + 10000 51 53 52 11 2 7

TABLE 2. SALMONELLA TYPHIMURIUM STRAIN TA1537. FDA COMPOUND F76-019 (Potassium Sodium Tartrate)
Compound	Metabolic	Micrograms of compound	Histidine revertants per plate
	Activation	added per plate		Experiment 1			Experiment 2
				28 February 1978			14 March 1978
					Average			Average
Negative control	−		17	24	16	6	3	5
	−		9	14		4	5
	+		12	5	11	11	3	7
	+		15	13		10	4
Positive controls
9-Aminoacridine	−	50	150	219	185	154
2-Anthramine	−	1	NT			3
	+	1	NT			41
F76-019	−	0,3	12	13	13
	−	3,3	10	19	15
	−	33,3	15	20	18	5	4	5
	−	100	17	7	12	6	2	4
	−	333,3	13	15	14	5	5	5
	−	1000	9	12	11	1	8	5
	−	3333,3	14	23	19	1	5	3
	−	10000	18	9	14	5	4	5
	+	0,3	14	14	14
	+	3,3	16	19	18
	+	33,3	15	16	16	6	12	9
	+	100	18	18	18	8	9	9
	+	333,3	17	10	14	7	8	8
	+	1000	13	11	12	11	4	8
	+	3333,3	14	12	13	6	3	5
	+	10000	13	24	19	8	4	6

TABLE 3. SALMONELLA TYPHIMURIUM STRAIN TA1538. FDA COMPOUND F76-019 (Potassium Sodium Tartrate)
Compound	Metabolic	Micrograms of compound	Histidine revertants per plate
	Activation	added per plate		Experiment 1			Experiment 2
				28 February 1978			14 March 1978
					Average			Average
Negative control	−		13	13	13	9	14	11
	−		12	13		9	10
	+		21	16	22	24	19	21
	+		28	23		18	21
Positive controls
2-Nitrofluorene	−	5	597	787	692	591
2-Anthramine	−	1	NT			19
	+	1	NT			89
F76-019	−	0,3	13	17	15
	−	3,3	14	11	13
	−	33,3	21	17	19	20	8	14
	−	100	5	14	10	14	4	9
	−	333,3	29	21	25	19	17	18
	−	1000	14	8	11	15	7	11
	−	3333,3	18	10	14	14	16	15
	−	10000	13	19	16	20	14	17
	+	0,3	9	32	21
	+	3,3	14	19	17
	+	33,3	26	37	32	29	23	26
	+	100	30	34	32	27	22	25
	+	333,3	31	23	27	28	21	25
	+	1000	29	20	25	25	22	24
	+	3333,3	21	27	24	25	25	25
	+	10000	44	24	34	27	15	21

TABLE 4. SALMONELLA TYPHIMURIUM STRAIN TA98. FDA COMPOUND F76-019 (Potassium Sodium Tartrate)
Compound	Metabolic	Micrograms of compound	Histidine revertants per plate
	Activation	added per plate		Experiment 1			Experiment 2
				28 February 1978			14 March 1978
					Average			Average
Negative control	−		28	21	24	19	18	15
	−		20	27		11	12
	+		37	38	36	18	37	27
	+		41	29		27	27
Positive controls
2-Nitrofluorene	−	5	345	373	359	359
2-Anthramine	−	2,5	39			18
	+	2,5	965			104
F76-019	−	0,3	26	17	22
	−	3,3	25	21	23
	−	33,3	23	19	21	31	11	21
	−	100	25	22	24	19	21	20
	−	333,3	26	18	22	15	17	16
	−	1000	20	23	22	15	27	21
	−	3333,3	22	33	28	19	14	17
	−	10000	12	15	14	17	13	15
	+	0,3	51	42	47
	+	3,3	30	37	34
	+	33,3	32	48	40	39	48	44
	+	100	27	35	31	36	35	36
	+	333,3	37	38	38	18	36	27
	+	1000	42	47	45	18	37	28
	+	3333,3	38	30	34	21	36	29
	+	10000	32	37	35	29	24	27

TABLE 6. ESCHERICHIA COLI WP2. FDA COMPOUND F76-019 (Potassium Sodium Tartrate)
Compound	Metabolic	Micrograms of compound	Tryptophan Revertants per plate
	Activation	added per plate		Experiment 1			Experiment 2
				28 February 1978			14 March 1978
					Average			Average
Negative control	−		65	45	50	33	36	33
	−		60	31		31	30
	+		55	53	57	57	45	40
	+		65	56		24	34
Positive controls
AF2	−	0,1	104	102	103	1581
2-Anthramine	−	10	72			NT*
	+	10	166
F76-019	−	0,3	43	54	49
	−	3,3	53	48	51
	−	33,3	39	55	47	42	28	35
	−	100	45	37	41	32	39	36
	−	333,3	51	39	45	39	43	41
	−	1000	42	30	36	39	29	34
	−	3333,3	44	52	48	23	48	36
	−	10000	57	48	53	29	49	39
	+	0,3	57	37	47
	+	3,3	52	53	53
	+	33,3	48	48	48	43	44	44
	+	100	53	48	51	50	66	58
	+	333,3	53	50	52	50	36	43
	+	1000	63	55	59	46	57	52
	+	3333,3	48	50	49	50	48	49
	+	10000	52	56	54	53	60	57

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Potassium sodium tartrate tested in the Ames test with and without metabolic activation on Salmonella strains TA 1535, TA 1537, TA98, TA 100 and E.coli strain WP2 did not reveal any significant increase in revertants attributable to the test concentrations tested, neither signs of toxicity (as seen by background lawn observation) up to a maximum concentration of 10 mg/plate. Thereby the sample is considered to be negative in the bacterial mutagenicity test performed.

Executive summary:

Potassium sodium tartrate tested in the Ames test with and without metabolic activation on Salmonella strains TA 1535, TA 1537, TA98, TA 100 and E.coli strain WP2 did not reveal any significant increase in revertants attributable to the test concentrations tested, neither signs of toxicity (as seen by background lawn observation) up to a maximum concentration of 10 mg/plate. Thereby the sample is considered to be negative in the bacterial mutagenicity test performed.