Bis(2-methoxyethyl) ether

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:

7 Sept - 12 Sept 1979

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Well performed study equivalent to current OECD Guidelines with quality inspection; E. coli not tested

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:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

E. coli not tested

GLP compliance:

no

Remarks:

study performed before GLP

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Bis(2-methoxyethyl) ether

EC Number:

203-924-4

EC Name:

Bis(2-methoxyethyl) ether

Cas Number:

111-96-6

Molecular formula:

C6H14O3

IUPAC Name:

1-methoxy-2-(2-methoxyethoxy)ethane

Constituent 2

Reference substance name:

Diglyme

IUPAC Name:

Diglyme

Constituent 3

Reference substance name:

Diethylenglycol-Dimethylether

IUPAC Name:

Diethylenglycol-Dimethylether

Details on test material:

Colorless liquid

Method

Species / strain

Species / strain / cell type:

other: Salmonella thyphimurium TA98, TA100, TA1535, TA1537, TA1538

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

0.005, 0.01, 0.1, 1.0, 5.0, 10.0, 25.0 and 50.0 µL/plate

Vehicle / solvent:

Distilled water

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

Distilled water

Positive controls:

yes

Positive control substance:

other: without S9 mix: TA98/1538 - 2-Nitrofluorene; TA100/1535 - Sodium azide; TA1537 - 9-Aminocaridine; with S9 mix: 2-Anthramine

Details on test system and experimental conditions:

- Dosage Selection
All tests are run at a minimum of 4 concentration. In the standard plate test, at least 6 dose levels of the test material, dissolved in a suitable solvent, are added to the test system. The standard test doses are 0.005, 0.01, 0.1, 1.0, 5.0 and 10.0 µL/plate.

- Mutagenicity Testing
the procedure is based on the Ames publication (1975) and performed as follows:

(1) Nonactivation Assay
To a sterile test tube placed in a 43°C water bath the following is added in order:
(a) 2.00 mL of 0.6% agar containing 0.05 mM Histidine and 0.05 mM Biotin
(b) 0.05 mL of a solution of Diethylene glycol dimethyl ether to give approximate dose
(c) 0.1 mL - 0.2 mL of indicator organisms
(d) 0.5 mL of 0.01M Phosphate buffer, pH 7.4
This mixture is swirled gently and then poured into minimal agar plates . After the top agar has set, the plates are incubated at 37°C for approx. 2 days. The number of his+ revertant colonies growing on the plates is counted.

(2) Activation assay
The activation assay is run concurrently with the nonactivation assay. The only difference is the addition of 0.5 mL of S9 mix (S9 supernatant from Aroclor 1254 induced adult male Sprague-Dawley rats) to the tubes in place of 0.5 mL of Phosphate buffer which is added in nonactivation assays. All other details are similar to the procedure for nonactivation assays.

- Control compounds
A negative control consisting of the solvent (distilled water) used for the test material is performed in all cases. For negative controls, step (b) of Nonactivation assays is replaced by 0.05 mL of the solvent. The negative controls are employed for each indicator strain and are performed in the absence and presence of S9 mix.
Specific positive control compounds known to revert each strain are also used in the assays:
(a) Nonactivation
- Sodium azide (1 µg/plate): TA100, TA1535
- 2-Nitrofluorene (10 µg/plate): TA98, TA1538
- 9-Aminocridine (50 µg/plate): TA1537

(b) Activation
- 2-Anthramine (2.5 µg/plate): all strains

Evaluation criteria:

(1) Strains TA1535, TA1537 and TA1538
If the solvent control value is within the normal range, a test material that produces a positive dose response over three times concentrations with the highest increase equal to three times the solvent control values will be considered to be mutagenic.

(2) strains TA98 and TA 100
If the solvent control vallue is within the normal range, a test material that produces a positive dose response over three concnetrations with the highest increase equal to twice the solvent control value for TA98 and TA 100 will be considered to be mutagenic.

(3) Reproducibility
If a test material produces a response in a single test that cannot be reproduced in additional runs, the initial positive test data lose significance.

The preceding criteria are not absolute, and other extenuating factors may enter into a final evaluation decision. However, these criteria will be applied to the majority of situations.

Statistics:

None

Results and discussion

Test results

Test results 1

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:

valid

True negative controls validity:

not specified

Positive controls validity:

valid

Test results 2

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:

valid

True negative controls validity:

not specified

Positive controls validity:

valid

Test results 3

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:

valid

True negative controls validity:

not specified

Positive controls validity:

valid

Test results 4

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:

valid

True negative controls validity:

not specified

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 1538

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

True negative controls validity:

not specified

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 1: Summary of mean number of revertants in Salmonella typhimurium with and without metabolic activation (negative and positive controls)

Concentration	TA 98		TA 100			TA1535					TA 1537
[µL/plate]	- S9 mix	+ S9 mix	- S9 mix		+ S9 mix	- S9 mix		+ S9 mix			- S9 mix	+ S9 mix
0	25	42	121		121	21		13			13	21
0.05	24	36	126		104	10		8			19	16
0.01	22	37	128		98	15		10			13	11
0.1	34	36	109		109	15		10			15	16
1.0	25	27	127		130	16		6			7	14
5.0 10.0 25.0 50.0	27 30 35 36	35 32 40 45	130 137 173 173		110 146 109 146	15 18 - -		14 5 - -			9 10 - -	9 14 - -
Positive:
9-Aminoacridine											518
Sodium azide			1433			1119
2-Nitrofluorene	1170
2-Anthramine		2109			2631			592				499
Concentration	TA 1538
[µL/plate]	- S9 mix	+ S9 mix
0	21	23
0.05	14	29
0.01	11	19
0.1	19	22
1.0	14	33
5.0 10.0 25.0 50.0	19 20 - -	36 22 - -
Positive:
9-Aminoacridine
Sodium azide
2-Nitrofluorene	1367
2-Anthramine
		1876

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results:
negative

Diethylene glycol dimethyl ether was not genotoxic in any of the bacterial reverse mutation assays (Ames test) performed with Salmonella th. TA98, TA100, TA1535, TA1537 and TA1538.

Executive summary:

Diethylene glycol dimethyl ether was examined for mutagenic activity in Salmonella th. The compound was tested directly and in the presence of liver microsomal enzyme preparations from Aroclor-induced rats.The dose range employed for the evaluation of this compound was from 0.005 to 50 µL/plate.

Diethylene glycol dimethyl ether exhibited slight toxicity in the activation assay with the strain TA1535 at 10 µL/plate.

The results of the tests conducted on Diethylene glycol dimethyl ether in the presence and absence of metabolic activation were all negative.