Alcohols, C16-18 (even numbered, C18-unsatd.), ethoxylated, and alcohols C20-22 (even numbered), sulfates, ammonium salts

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

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

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study

Remarks:

Comparable to guideline study without detailed documentation (publication). Read-across is justified due to the fact that the test item contains siginificant amounts of long-chain fatty alcohols (Octadecan-1-ol, CAS 112-92-5, icosan-1-ol, CAS 629-96-9 and docosan-1-ol, CAS 661-19-8). Toxicological data on Docosan-1-ol, CAS 661-19-8 is considered as representative for this set of long chain, linear fatty alcohols.

Data source

Reference

Reference Type:

review article or handbook

Title:

Unnamed

Year:

2002

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

yes (only 1000 PCEs per animal scored for micronuclei) Principles of method if other than guideline

Principles of method if other than guideline:

Study according to protocol very similar to OECD guideline 474.

GLP compliance:

not specified

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

Docosan-1-ol

EC Number:

211-546-6

EC Name:

Docosan-1-ol

Cas Number:

661-19-8

Molecular formula:

C22H46O

Constituent 2

Reference substance name:

behenyl alcohol

IUPAC Name:

behenyl alcohol

Test material form:

solid: compact

Details on test material:

- Name of test material (as cited in study report): behenyl alcohol
- Substance type: colourless, waxy solid
- Physical state: solid
- Analytical purity: no data, but for repeated dose toxicity (oral), 98%
- Impurities (identity and concentrations): no data
- Composition of test material, percentage of components: no data
- Isomers composition: no data
- Purity test date: no data
- Lot/batch No.: no data, but for repeated dose toxicity (oral), obtained from Condea, Germany
- Expiration date of the lot/batch: no data
- Stability under test conditions: no data
- Storage condition of test material: no data
- Other:
- Melting point: 70 deg C
- Soluble in: ethanol and chloroform
- Insoluble in: water

Test animals

Species:

mouse

Strain:

NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: BRL Tierfarm Fullinsdorf, Switzerland
- Age at study initiation: >=10 weeks
- Weight at study initiation: no data
- Assigned to test groups randomly: no data
- Fasting period before study: 18 hours, but continued to receive water ad libitum
- Housing: Markrolon Type 1 cages with wire mesh tops and granulated soft wood bedding
- Diet (e.g. ad libitum): standard pellet diet, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: no data

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21
- Humidity (%): not regulated
- Air changes (per hr): no data
- Photoperiod (hrs dark / hrs light): 12 / 12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: polyethylene glycol
- Justification for choice of solvent/vehicle: no data
- Concentration of test material in vehicle: no data [calculated: 5, 15 and 50 mg/ml]
- Amount of vehicle (if gavage or dermal): 10 ml/kg bw
- Lot/batch no. (if required): no data
- Purity: no data

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: few details; test material suspended in vehicle

Duration of treatment / exposure:

single administration

Frequency of treatment:

single administration

Post exposure period:

none

Doses / concentrations

Remarks:

Doses / Concentrations:
50, 150, 500 mg/kg bw
Basis:
nominal conc.

No. of animals per sex per dose:

6

Positive control(s):

cyclophosphamide
- Justification for choice of positive control(s): no data
- Route of administration: presumably oral gavage
- Doses / concentrations: 40 mg/kg bw

Examinations

Tissues and cell types examined:

bone marrow cells

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: based on previous study - 500 mg/kg bw estimated to be the "maximum attainable dose"
TREATMENT AND SAMPLING TIMES (in addition to information in specific fields): 24, 48 and 72 hours after dosing

DETAILS OF SLIDE PREPARATION: femurs removed, marrow flushed out with foetal calf serum, cell suspension centrifuged and supernatant discarded, small drop of cell pellet spread on slide, air dried, stained with May-Grunwald, mounted; 1 slide/sample

METHOD OF ANALYSIS: 1000 polychromatic erythrocytes (PCEs) scored for micronuclei; polychromatic:normochromatic (PCE:NCE) ratio scored

OTHER: only 5/sex per dose level evaluated

Evaluation criteria:

To be considered positive, either a statistically significant dose-related increase in the number of micronucleated PCEs or a reproducible, statistically significant positive response for at least one dose level.

Statistics:

Mann-Whitney test

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

not specified

Remarks:

presumably toxic at >500 mg/kg bw

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- Dose range: no data
- Solubility: no data
- Clinical signs of toxicity in test animals: no data
- Evidence of cytotoxicity in tissue analyzed: no data
- Rationale for exposure: no data
- Harvest times: no data
- High dose with and without activation: no data
- Other: presumably toxic above 500 mg/kg bw since this maximum dose was chosen for the main study on the basis of the results of the range-finding study

RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): 0.03-0.09% for vehicle controls, 0.04-0.10% for test material treated, 0.71% for positive control
- Ratio of PCE/NCE (for Micronucleus assay): 1.05-1.27 for vehicle controls, 0.98-1.55 for test material treated, 0.93 for positive control
- Appropriateness of dose levels and route: appropriate (top dose was apparently the maximum tolerated dose, oral route relevant to humans)
- Statistical evaluation: no statistically significant increases in the frequency of micronuclei in mice treated with the test material; statistical significance not presented for positive controlAny other information on results incl. tables

Any other information on results incl. tables

Toxicity unclear, but possibly one male and one female mouse [per group?] died either spontaneously or due to gavage error.

Table 1: Results of micronucleus assay 24 hour sampling time

Treatment	Suspending agent	Low dose	Mid dose	High dose
Concentration mg/kg bw	0	40	50	150
Harvest time	24	24	24	24
Micronucleated PCE (%)	0.03	0.71	0.07	0.08
Ratio PCE/NCE	1.27	0.93	0.98	1.07

Table 2: Results of micronucleus assay 48 hour sampling time

Treatment	Suspending agent	Test substance	Test substance	Test substance
Concentration mg/kg bw	0	50	150	500
Harvest time	48	48	48	48
Micronucleated PCE (%)	0.09	0.1	0.04	0.05
Ratio PCE/NCE	1.05	1.06	1.01	1.23

Table 3 Results of micronucleus assay 72 hour sampling time

Treatment	Suspending agent	Low dose	Mid dose	High dose
Concentration mg/kg bw	0	50	150	500
Harvest time	72	72	72	72
Micronucleated PCE (%)	0.09	0.09	0.05	0.07
Ratio PCE/NCE	1.41	1.33	1.55	1.46

PCE = polychromatic erythrocytes

NCE = normochromatic erythrocytes

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative

Executive summary:

Behenyl alcohol did not increase the incidence of micronuclei in mouse bone marrow cells after a single oral gavage dose of up to 500 mg/kg bw.