Alcohols, C12-14

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:

other: published data

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1992

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

no

GLP compliance:

not specified

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

Dodecan-1-ol

EC Number:

203-982-0

EC Name:

Dodecan-1-ol

Cas Number:

112-53-8

Molecular formula:

C12H26O

IUPAC Name:

dodecan-1-ol

Details on test material:

- Name of test material (as cited in study report): 1-Dodecanol (Lorol C12-99) - Substance type: fatty alcohol- Physical state: colourless liquid, pasty- Analytical purity: 100% (active substance)- Lot/batch No.: A 06502:91- Expiration date of the lot/batch: Apr-1992- Stability under test conditions: not given (pure: at least 6 months)- Storage condition of test material: at room temperature, dark- Other:Production date: 24-Oct-1991Receipt of test material: 26-Nov-1991

Test animals

Species:

mouse

Strain:

other: albino mice, CFW 1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Winkelmann
- Age at study initiation: 7-8 weeks
- Weight at study initiation: males 21-27 g, females 21-26 g
- Assigned to test groups randomly: yes, under following basis: allocated to treatment groups according to randomization table generated by computer programme or manually
- Fasting period before study: yes, overnight until 3-4 hours after dosing
- Housing: males, 1/cage, macrolon cages type I; females, <=3/cage, macrolon cages type II; filled with clean softwood bedding
- Diet (e.g. ad libitum): standard animal diet, Altromin No. 1314, ad libitum
- Water (e.g. ad libitum): tap water, ad libitum
- Acclimation period: >=6 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 25 +- 3 (occasionally 20-25)
- Humidity (%): 40 - 50 (occasionally 45-70)
- Air changes (per hr): no data, except "air-conditioned room"
- Photoperiod (hrs dark / hrs light): 12 / 12

IN-LIFE DATES (main study): From: 25-Feb-1992 To: 28-Feb-1992

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: arachis oil
- Justification for choice of solvent/vehicle: test material easily soluble at required concentration
- Concentration of test material in vehicle: not stated, but provided a dose level of 5000 mg/kg bw, so 500 mg/ml
- Amount of vehicle (if gavage or dermal): 10 ml/kg bw (main study), 20 ml/kg bw (range finding study)
- Lot/batch no. (if required): no data
- Purity: no data

Details on exposure:

PREPARATION OF DOSING SOLUTIONS: 500 mg/ml in arachis oil (main study)

Duration of treatment / exposure:

single administration

Frequency of treatment:

single administration

Post exposure period:

evaluated at 24, 48, 72 hours after administration

Doses / concentrations

Remarks:

Doses / Concentrations:
5000 mg/kg bw
Basis:
nominal conc.

No. of animals per sex per dose:

6

Control animals:

yes, concurrent vehicle

Positive control(s):

cyclophosphamide
- Justification for choice of positive control(s): not stated
- Route of administration: oral
- Dose: 20 mg/kg bw

Examinations

Tissues and cell types examined:

bone marrow

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: maximum tolerated dose, based on range-finding study (effects seen at 5000 mg/kg were piloerection only)

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): single administration, animals sacrificed 24, 48 and 72 hours after administration

DETAILS OF SLIDE PREPARATION: bone marrow collected from femurs, using foetal calf serum applied via a syringe, into a siliconised centrifuge tube; after centrifugation at 1000 rpm and removal of all but one drop of supernatant, cells of sediment carefully mixed; drop of cell suspension placed on clean, degreased microscope slide and immediately spread; 3 slides/animal; slides air dried at least overnight; stained with Giemsa; air dried and dipped in xylol for 3 minutes

METHOD OF ANALYSIS: 1 slide/animal chosen and given a random code; microscopic evaluation of slides from 5 males and 5 females per treatment group at 1000x magnification; number of micronucleated cells counted in 1000 polychromatic erythrocytes (PCEs)/animal; ratio of
polychromatic to normochromatic erythrocytes determined by counting and differentiating the first 1000 erythrocytes

OTHER: means and standard deviations calculated

Evaluation criteria:

Statistically significant (p<0.05) increase in PCE compared to controls at any sampling time in either sex
Acceptability of test: positive controls induced statistically significant increase in frequency of micronucleated PCEs; solvent control incidence of micronuclei should reasonably fall within historical control range for the testing facility.

Statistics:

Method used: Kastenbaum & Bowman

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Remarks:

piloerection for 8 hours after administration; no mortality

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- Dose range: 5000 mg/kg bw
- Solubility: used at 250 mg/ml
- Clinical signs of toxicity in test animals: piloerection
- Evidence of cytotoxicity in tissue analyzed: not examined
- Rationale for exposure: based on limit test in rats in which acute oral LD50 was >5000 mg/kg bw
- Harvest times: animals observed for 3 days
RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): no statistically significant increase in micronucleus frequency in any treatment group
- Ratio of PCE/NCE (for Micronucleus assay): treated groups similar to controls
- Appropriateness of dose levels and route: maximum tolerated dose of 5000 mg/kg bw used; guideline recommends maximum dose of 2000 mg/kg bw; oral route selected "taking into account the possible route of human exposure during manufacture, handling and use"
- Statistical evaluation: no statistically significant increases in micronuclei in treated groups of either sex; positive control produced a statistically significant increase in micronuclei
- Control incidence of micronuclei: not reported but presumably therefore within historical control range

Any other information on results incl. tables

Lorol 12 did not increase the frequency of micronucleated erythrocytes or the PCE:NCE ratio in mice at any time interval after treatment (24, 48 or 72 hours) at dose levels up to 5000 mg/kg bw when compared to vehicle controls.

Mean values per group  in the micronucleus test with 1-Dodecanol (Lorol C12-99)

a) Number of micronucleated cells per 1000 polychromatic erythrocytes (PCE)

b) Ratio of polychromatic to normochromatic erythrocytes (PCE/NCE)

Treatment group;  (sampling time)	Species and sex	Dose mg/kg	Micronucleated cells 1000 PCE		Ratio of PCE/NCE
			Mean	Range	Mean	Range
Negative control (24 hours) arachis oil	male mice	10 ml/kg	3.60	0 - 9	1.11	0.80 - 1.31
	female mice	10 ml/kg	2.00	0 - 4	1.34	1.02 - 1.07
Positive control (24 hours) cyclophosphamide	male mice	20	13.40	10 - 16	1.21	0.90 - 1.72
	female mice	20	10.80	7 - 14	0.95	0.67 - 1.28
1-Dodecanol (Lorol C12-99)
Limit dose (24 hours)	male mice	5000	2.60	0 - 5	1.08	0.94 - 1.26
	female mice	5000	2.40	2 - 3	1.01	0.90 - 1.18
Limit dose (48 hours)	male mice	5000	3.00	1 - 4	0.89	0.48 - 1.16
	female mice	5000	2.00	0 - 5	1.18	0.90 - 1.68
Limit dose (72 hours)	male mice	5000	2.60	2 - 4	1.65	0.91 - 2.14
	female mice	5000	1.60	0 - 4	1.33	1.08 - 1.55

Applicant's summary and conclusion

Conclusions:

Interpretation of results: negative
Dodecan-1-ol has been tested a reliable study, conducted according to OECD guideline 474, no genotoxicity was seen in mice after a single oral dose of 5000 mg/kg bw. . The test substance, dodecan-1-ol is closely related to the registration substance, Alcohols, C12-14 and it is considered that read-across is valid.Dodecan-1-ol (C12) is supporting substance for Alcohols,C12-C14 and the main component.