Reaction products of monoethanolamine and boric acid (1:1)

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:

16 Jun 2017 - 10 Jul 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

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

1

Reference substance name:

3-hydroxypropan-1-aminium; tetrahydroxyspiro[5.5]pentaboroxan-6-uide

EC Number:

701-025-6

Cas Number:

26038-87-9

Molecular formula:

C2H7NO.BH3O3

IUPAC Name:

3-hydroxypropan-1-aminium; tetrahydroxyspiro[5.5]pentaboroxan-6-uide

Test material form:

liquid

Details on test material:

Appearance: Clear colourless liquid
Purity/Composition: UVCB
Test item storage: At room temperature protected from light
Stable under storage conditions until: 31 March 2018 (expiry date)

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:

Rat liver microsomal enzymes (S9 homogenate), prepared from male Sprague Dawley rats that had been injected intraperitoneally with Aroclor 1254 (500 mg/kg body weight).

Test concentrations with justification for top dose:

Dose-range Finding Test
Selection of an adequate range of doses was based on a dose-range finding test with the strains TA100 and the WP2uvrA, both with and without S9-mix. Eight concentrations, 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 µg/plate were tested in triplicate.

The highest concentration of the test item used in the subsequent mutation assays was 5000 μg/plate. At least five different doses (increasing with approximately half-log steps) of the test item were tested in triplicate in each strain in the absence and presence of S9-mix.

Vehicle / solvent:

The vehicle of the test item was Milli-Q water (Millipore Corp., Bedford, MA., USA).

Controls

Controls 1

Untreated negative controls:

yes

Remarks:

Milli-Q water

Controls 2

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

2-nitrofluorene

sodium azide

methylmethanesulfonate

other: ICR-191 (Sigma), 2-aminoanthracene (2AA)

Details on test system and experimental conditions:

Salmonella typhimurium bacteria and Escherichia coli bacteria

Rationale for test conditions:

Recommended test system in international guidelines (e.g. OECD, EC).

Results and discussion

Test results

Test results 1

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

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 2

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

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 3

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

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 4

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

Untreated negative controls validity:

valid

Positive controls validity:

valid

Test results 5

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

Untreated negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Dose-Range Finding Test: Mutagenic Response of MEA Polyborate 1:1 in the Salmonella typhimurium Reverse Mutation Assay and in the Escherichia coli Reverse Mutation Assay

Direct plate assay

Dose (µg/plate)	Mean number of revertant colonies/3 replicate plates (±S.D.) with one Salmonella typhimurium and one Escherichia colistrain.
	TA100	WP2uvrA

Without S9-mix

Positive control	722	±	32		1272	±	76
Solvent control	101	±	7		19	±	6
1.7	99	±	7		19	±	1
5.4	108	±	5		18	±	3
17	101	±	25		25	±	6
52	117	±	9		23	±	0
164	111	±	29		21	±	5
512	100	±	16		22	±	8
1600	110	±	6		20	±	5
5000	128	±	11	^n NP	19	±	6	^n NP

With S9-mix

Positive control	475	±	106		420	±	13
Solvent control	92	±	6		29	±	10
1.7	107	±	15		27	±	4
5.4	95	±	12		25	±	4
17	110	±	12		28	±	8
52	111	±	16		29	±	11
164	112	±	14		24	±	5
512	107	±	9		27	±	6
1600	101	±	11		27	±	2
5000	126	±	17	^n NP	38	±	4	^n NP

NP	No precipitate
n	Normal bacterial background lawn

Experiment 1: Mutagenic Response of MEA Polyborate 1:1 in the Salmonella typhimurium Reverse Mutation Assay

Direct plate assay

Dose (µg/plate)	Mean number of revertant colonies/3 replicate plates (±S.D.) with different strains of Salmonella typhimurium.
	TA1535	TA1537	TA98

 Without S9-mix

Positive control	790	±	47		985	±	99		1193	±	86
Solvent control	11	±	2		6	±	2		16	±	3
52	16	±	3		6	±	2		11	±	6
164	11	±	4		4	±	1		15	±	5
512	11	±	7		6	±	2		25	±	5
1600	11	±	4		6	±	2		16	±	9
5000	6	±	4	^n NP	4	±	1	^n NP	16	±	2	^n NP

 

With S9-mix

Positive control	328	±	21		546	±	104		1022	±	61
Milli-Q water	10	±	3		8	±	4		24	±	11
52	13	±	2		7	±	2		29	±	5
164	10	±	2		6	±	3		25	±	9
512	12	±	1		7	±	3		30	±	6
1600	13	±	3		5	±	0		24	±	3
5000	7	±	2	^n NP	8	±	4	^n NP	21	±	2	^n NP

 

NP	No precipitate
n	Normal bacterial background lawn

Experiment 2: Mutagenic Response of MEA Polyborate 1:1 in theSalmonella typhimuriumReverse Mutation Assay and in the Escherichia coli Reverse Mutation Assay

Pre-incubation assay

Dose (µg/plate)	Mean number of revertant colonies/3 replicate plates (±S.D.) with different strains ofSalmonella typhimuriumand oneEscherichia colistrain.
	TA1535	TA1537	TA98	TA100	WP2uvrA

Without S9-mix

Positive control	886	±	21		1183	±	75		1211	±	169		708	±	171		202	±	35
Solvent control	7	±	2		9	±	1		20	±	5		118	±	4		36	±	9
52	13	±	1		8	±	2		34	±	21		114	±	13		25	±	10
164	12	±	2		8	±	4		19	±	7		104	±	51		29	±	4
512	15	±	5		8	±	3		18	±	4		112	±	20		31	±	4	^i
1600	9	±	6		11	±	6		12	±	4		149	±	8		31	±	13
5000	9	±	3	^n SP	5	±	2	^n SP	15	±	5	^n SP	179	±	17	^n SP	38	±	17	^n SP

 

With S9-mix

Positive control	217	±	3		177	±	16		495	±	86		1008	±	343		541	±	13
Milli-Q water	19	±	5		4	±	0		26	±	2		118	±	21		42	±	9
52	8	±	1		8	±	2		28	±	13		116	±	10		41	±	6
164	11	±	4		6	±	5		28	±	2		135	±	27		61	±	7
512	10	±	2		8	±	4		25	±	7		145	±	27		54	±	9
1600	13	±	1		10	±	8		23	±	9		148	±	11		48	±	9
5000	7	±	2	^n SP	5	±	3	^n SP	21	±	4	^n SP	189	±	18	^n SP	52	±	5	^n SP

 

SP	Slight Precipitate
i	Plate infected
n	Normal bacterial background lawn

 

Applicant's summary and conclusion

Conclusions:

Based on the results of a GLP OECD 471 study it is concluded that MEA Polyborate 1:1 is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.