Reaction mass of octadecyl heptanoate and octadecyl octanoate

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:

experimental study

Adequacy of study:

weight of evidence

Study period:

13 Aug - 12 Oct 1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Guideline study with acceptable restrictions. There was no negative control for the 48-h sampling time, the purity of the test substance was not specified.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1993

Report date:

1993

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Version / remarks:

adopted Sep 2014

Deviations:

yes

Remarks:

no negative control group for the 48-h sampling time; purity of the test substance not specified

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayerisches Staatsministerium für Arbeit, Familie und Sozialordnung, München

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

octadecyl heptanoate; octadecyl octanoate

EC Number:

915-334-0

Molecular formula:

C25H50O2 C26H52O2

IUPAC Name:

octadecyl heptanoate; octadecyl octanoate

Details on test material:

- Name of test material (as cited in study report): stearyl heptanoate (and) stearyl caprylate
- Physical state: waxy, colourless solid
- Analytical purity: no data
- Lot/batch No.: A:257032
- Storage condition of test material: at room temperature in the dark

Test animals

Species:

mouse

Strain:

other: Crl:NMRI BR

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Wiga, Sulzfeld, Germany
- Age at study initiation: adult (approximately 3 months)
- Weight at study initiation: 21 g ± 15%
- Assigned to test groups randomly: yes
- Housing: animals were housed in groups of 5 in Macrolon cages
- Diet: rat/mouse/hamster feed, pellets (Eggersmann Futtermittelwerk, Rinteln, Germany), ad libitum
- Water: tap water, ad libitum
- Acclimation period: at least 6 - 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 2
- Humidity (%): 55 ± 10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: corn oil
- Amount of vehicle (if gavage or dermal): 10 mL/kg bw

Duration of treatment / exposure:

not applicable

Frequency of treatment:

single treatment

Post exposure period:

24 h after treatment (500 and 1500 mg/kg bw group)
24 and 48 h after treatment (5000 mg/kg bw group)
24 h after treatment (control groups)

Doses / concentrations

Remarks:

Doses / Concentrations:
500, 1500 and 5000 mg/kg bw
Basis:
actual ingested

No. of animals per sex per dose:

5

Control animals:

other: yes, concurrent vehicle. 2 negative control groups were used for the 24-hour sampling time and none for the 48-hour sampling time; no justification was given.

Positive control(s):

cyclophosphamide
- Route of administration: oral, gavage
- Doses / concentrations: 40 mg/kg bw

Examinations

Tissues and cell types examined:

Tissue: bone marrow
Cell type: bone marrow cells

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION:
A range-finding study was performed to identify the maximum tolerated dose.

DETAILS OF SLIDE PREPARATION:
Slides were fixed with methanol for 5 minutes and stained with a May-Grünwald and Giemsa solution. Two slides per animal were prepared.

METHOD OF ANALYSIS:
A total of 1000 polychromatic erythrocytes were examined per slide and the number of micronucleated cells were recorded. The ratio of polychromatic erythrocytes to normochromatic (mature) erythrocytes was calculated for a sample of 1000 cells. All slides were randomised before analysis.

Evaluation criteria:

The frequency of micronuclei in the polychromatic erythrocytes and the ratio of polychromatic erythrocytes to normochromatic (mature) erythrocytes were compared with the negative control groups, the positive control group and the historical data.

Statistics:

The data were analysed using a likelihood ratio test. Individual animal results were used as data points in the analysis. The set of micronuclei in polychromatic erythrocytes among the control groups were compared with the set of each treatment time.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

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
- Clinical signs of toxicity in test animals: there were no signs of toxicity during the 48-h observation period
- Rationale for exposure: the dose was selected based on the maximum dose level recommended by regulatory bodies

RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): the number of polychromatic erythrocytes with or without micronuclei and the number of normochromatic erythrocytes in the treatment groups at different sampling times did not differ significantly from the number for the vehicle control groups and historical vehicle control data (see Table 1 - 4 under 'Any other information on results incl. tables').
- Ratio of PCE/NCE (for Micronucleus assay): the ratio for the treatment groups at different sampling times did not differ significantly from the ratio for the vehicle control groups and historical vehicle control data (see Table 1 - 4 under 'Any other information on results incl. tables').
- Other: no clinical signs were observed during the 48-h observation period.

Any other information on results incl. tables

Table 1: Results of the in vivo micronucleus assay in male animals

 

			Mean PCEs / 1000 NCEs at sampling time		Total micronuclei per 1000 PCEs at sampling time
Group	Number of animals	Dose [mg/kg bw]	24 h	48 h	24 h	48 h
Vehicle control I (corn oil)	5	0	498.5 ± 54.4	n.d.	2.20 ± 1.10	n.d.
Vehicle control II (corn oil)	5	0	507.4 ± 78.6	n.d.	2.20 ± 1.10	n.d.
Positive control (cyclophosphamide)	5	40	555.0 ± 116.2	n.d.	41.8 ± 7.92	n.d.
Test substance	5	500	546.4 ± 125.5	n.d.	2.0 ± 1.22	n.d.
Test substance	5	1500	545.0 ± 81.9	n.d.	1.20 ± 0.84	n.d.
Test substance	5	5000	553.0 ± 68.9	582.2 ± 26.1	2.80 ± 1.10	2.20 ± 1.30

n.d. = not determined

 

 

Table 2: Results of the in vivo micronucleus assay in female animals

 

			Mean PCEs / 1000 NCEs at sampling time		Total micronuclei per 1000 PCEs at sampling time
Group	Number of animals	Dose [mg/kg bw]	24 h	48 h	24 h	48 h
Vehicle control I (corn oil)	5	0	578.6 ± 39.9	n.d.	2.40 ± 1.14	n.d.
Vehicle control II (corn oil)	5	0	557.6 ± 61.6	n.d.	2.40 ± 2.70	n.d.
Positive control (cyclophosphamide)	5	40	557.8 ± 73.6	n.d.	29.2 ± 11.9	n.d.
Test substance	5	500	584.8 ± 70.6	n.d.	1.0 ± 1.41	n.d.
Test substance	5	1500	618.6 ± 45.7	n.d.	2.20 ± 1.64	n.d.
Test substance	5	5000	549.6 ± 88.6	526.4 ± 59.4	1.0 ± 0.71	2.20 ± 1.30

n.d. = not determined

 

Table 3: PCE/NCE ratio males and females

Treatment group	Dose [mg/kg]	Sampling time [h]	PCE/NCE ratio
Vehicle control I (corn oil)	0	24	1.20 ± 0.29
Vehicle control II (corn oil)	0	24	1.19 ± 0.35
Test substance	500	24	1.40 ± 0.50
Test substance	1500	24	1.46 ± 0.46
Test substance	5000	24	1.29 ± 0.39
Test substance	5000	48	1.27 ± 0.24
Positive control (Cyclophosphamide)	40	24	1.24 ± 0.53

 

Table 4: Historical data, vehicle controls

Parameter	Mean	Normal range
Number of polychromatic erythrocytes with micronuclei per 1000 polychromatic erythrocytes
Male	1.95 ± 1.02	< 3.99
Female	1.98 ± 0.76	< 3.50
Male and female	-
Ratio of polychromatic to normochromatic erythrocytes (1000)
Male	1.12 ± 0.34	0.44 - 1.88
Female	1.13 ± 0.20	0.73 - 1.53
Male and female	1.13 ± 0.27	0.59 - 1.67

 

   

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative