Terpenes and terpenoids, turpentine-oil, α-pinene fraction, oligomers

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:

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

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Materials and methods

Test guideline

Guideline:

other: Micronucleus analysis in NTP toxicity studies

GLP compliance:

yes

Type of assay:

micronucleus assay

Test material

Test material information

Constituent 1

Reference substance name:

Alpha-pinene

IUPAC Name:

Alpha-pinene

Constituent 2

Reference substance name:

Pin-2(3)-ene

EC Number:

201-291-9

EC Name:

Pin-2(3)-ene

Cas Number:

80-56-8

IUPAC Name:

2,6,6-trimethylbicyclo[3.1.1]hept-2-ene

Details on test material:

Chemical name: alpha-pinene
CASRN: 80-56-8

Test animals

Species:

mouse

Strain:

B6C3F1

Sex:

male/female

Administration / exposure

Route of administration:

inhalation: aerosol

Vehicle:

Air

Details on exposure:

Start date: 2012/07/25 male
Start date: 2012/07/25 female
Sample collection time: 24hr

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

INHAL x 5

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 25, 50, 100, 200, 400 ppm
Basis:
other: inhalation

No. of animals per sex per dose:

5

Control animals:

yes

Examinations

Tissues and cell types examined:

Peripheral Blood

Statistics:

Official NTP Protocol

Data presented are the mean number of micronucleated cells per 1,000 cells for each treatment group. A positive trend test is one in which the P value is equal to or less than 0.025. For the slide-based micronucleus data, the micronucleus frequency in any dose group is considered significantly elevated over the control group if the P value is equal to or less than 0.025 divided by the number of chemical-treatment groups. Thus, if the number of treated groups is 3, then the required pairwise P value is 0.008. This adjustment in the pairwise P value is a correction for multiple comparisons of the same data.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Any other information on results incl. tables

Start Date	Sample Collection Time	Sex	Cell	Methodology Used	Dosing Regimen	Route	Trend Test P-Value
07/25/2005	24 Hours	Male	NCE	Slide Scoring	INHAL x 5, 13 Weeks	Inhalation	0.742
				Dose (ppm)	Number of Animals Scored	Mean MN-NCE/1000 NCE ± SEM	Pairwise P
Vehicle Control		Air		0	5	1.60 ± 0.33
Test Chemical		alpha-Pinene		25	5	1.80 ± 0.30	0.3657
				50	5	1.90 ± 0.53	0.3059
				100	5	2.10 ± 0.43	0.2053
				200	5	1.90 ± 0.29	0.3059
				400	5	1.40 ± 0.40	0.6426
A82162 - 2
Start Date	Sample Collection Time	Sex	Cell	Methodology Used	Dosing Regimen	Route	Trend Test P-Value
07/26/2005	24 Hours	Female	NCE	Slide Scoring	INHAL x 5, 13 Weeks	Inhalation	0.899
				Dose (ppm)	Number of Animals Scored	Mean MN-NCE/1000 NCE ± SEM	Pairwise P
Vehicle Control		Air		0	5	1.40 ± 0.19
Test Chemical		alpha-Pinene		25	5	2.10 ± 0.43	0.1182
				50	5	1.80 ± 0.25	0.2396
				100	5	1.70 ± 0.44	0.2949
				200	5	1.70 ± 0.30	0.2949
				400	5	1.10 ± 0.19	0.7259

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negative
The Micronucleus peripheral blood study conducted on Alpha-pinene showed no genotoxicity under the conditions of this study.

Executive summary:

A Peripheral blood micronucleus test study test was conducted as part of a NTP 13 week inhalation study on B6C3F1 female and male mice.

At the end of the treatment period, blood sample 13-week exposure period a blood sample was obtained from male and female mice. Sample collection time was typically between 0 (in the case of continuous exposures) and 24 hours (in the case of single daily exposures). 1,000 to 10,000 mature erythrocytes (normochromatic erythrocytes or NCEs) were scored per animal for presence of micronuclei. The percent PCE (immature erythrocytes) was determined in the blood as a measure of chemical-induced toxicity to the bone marrow. All data are analyzed separately for male and female mice. The results of the study showed no genetic toxicity under the test conditions.