Amides, C16-18 (even numbered) saturated and C18 unsaturated, N,N'-[[(2-hydroxyethyl)imino]di-2,1-ethanediyl]bis-, ethoxylated (1-2.5 mol)

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-03-25 until 2013-09-30

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: GB/T 21773-2008 Chemicals

GLP compliance:

yes

Type of assay:

other: mammalian bone marrow cells

Test material

Test material information

Constituent 1

Reference substance name:

Amides, C16-18 and C18-unsatd., N,N'-[[(2-hydroxyethyl)imino]di-2,1-ethanediyl]bis-,ethoxylated, lactates (salts)

Cas Number:

1434133-00-2

Molecular formula:

C47H96N3O5+ C3H5O3-

IUPAC Name:

Amides, C16-18 and C18-unsatd., N,N'-[[(2-hydroxyethyl)imino]di-2,1-ethanediyl]bis-,ethoxylated, lactates (salts)

Test material form:

solid: bulk

Details on test material:

Batch ST02697671

Specific details on test material used for the study:

Lot: ST02697671
purity: 86.8%

Test animals

Species:

mouse

Strain:

other: KM mice

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Changsa Tianqin biotechnology Ltd.
- Age at study initiation:
- Weight at study initiation:
- Assigned to test groups randomly: [no/yes, under following basis: ]
- Fasting period before study:
- Housing: SPF barrier environment
- Diet (e.g. ad libitum):
- Water (e.g. ad libitum):
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-25
- Humidity (%): 54-60
- Air changes (per hr):
- Photoperiod (hrs dark / hrs light):

IN-LIFE DATES: From: To:

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

distilled water

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:
weight 3.75 g test substance and add distilled water to 30 mL to prepare the stock solution with the concentration of 125 mg/mL for high dose group (2500 mg/kg). Then draw 4.0 mL and 8.0 mL high dose group solution, add distilled water to 16 mL, to prepare stock solution for low dose group (625 mg/kg) and medium dose group (1259 mg/kg) animals respectively. The concentrations were 31.2, 62.5 mg/mL respectively.

Duration of treatment / exposure:

The test substance was administered to mice at a volume of 0.2 mL/10 g bw by gavage twice during the period of 24h (at the beginning and at the 24h respectively). The solvent control group was treated by the same method. The positive control group was treated with cyclophosphamid (0.2%) by celiac injection at a volume of 0.15 ml/10 g bw.

Frequency of treatment:

twice, at 0 and 24 hours

Post exposure period:

18 hours

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

2 500 mg/kg bw (total dose)

Doses / concentrations 2

Dose / conc.:

1 250 mg/kg bw (total dose)

Doses / concentrations 3

Dose / conc.:

625 mg/kg bw (total dose)

Doses / concentrations 4

Dose / conc.:

30 mg/kg bw (total dose)

Remarks:

positive control, Cyclophosphamide

No. of animals per sex per dose:

5

Control animals:

yes

yes, concurrent vehicle

Positive control(s):

cyclophosphamide
- Route of administration: celiac injection
- Doses / concentrations: 30m/kg bw

Examinations

Tissues and cell types examined:

bone marrow cells

Details of tissue and slide preparation:

Bone marrow cells were harvested from the sternum and cell smears were prepared and stained according to conventional procedures. The region of cell-completely, well-distributed and well-colored up was selected and examined under a microscope. 2000 polychromatic erythrocytes per animal were scored for the incidence of micronclei. The ratio of PCEs to NCEs was also determined for each animal by counting a total of 200 erythrocytes, as an indication of cytotoxicity to the bone marrow cells.

Statistics:

The micronucleated PCE ratios for all dose-groups and solvent control group were analyzed using the poisson distribution and that of positive control group was handled with Gaussion distribution. Each sex was analyzed respectively. The parameters and data were analyzed statistically by SPSS 13.0.

Results and discussion

Test results

Key result

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Positive controls validity:

valid

Additional information on results:

No clinical signs were observed in any test animal. And no death occurred in any test animal during the period of the test.
The incidence of MNPCE in each treatment group were summarized Table 1. The incidence of MNPCE in solventr control groups were 1.4% and 1.6% for female and male mice respectively. The incidence of MNPCE were 1.0%, 1.7%, 1.5% for female mice and 0.8%, 1.3%, 0.9% for male mice in low, medium and high dose groups respectively, there were no significant differences compared with the solvent control (p>0.05). The incidence of MNPCE in positive control group were 24.9% and 22.6% for female and male mice respectively, there were significant differences compared with the solvent control group (p<0.01)
The result indicated, that the test substance could not increase the micronuclear rates of KM mice obviously.

Applicant's summary and conclusion

Conclusions:

It was concluded that the test substance was negative in the mammalian erythrocyte micronucleus test in vivo.