Glycerol trinitrate

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1.3.1975-25.2.1976

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to Guideline study study with acceptable restrictions.

Data source

Reference

Reference 1

Reference Type:

study report

Title:

Unnamed

Year:

1976

Report date:

1976

Reference 2

Reference Type:

publication

Title:

The development of a human amnion strain of cells.

Author:

Fernandes, M.V.

Year:

1956

Bibliographic source:

Texas Repts. Biol. Med., 16, 48.

Reference 3

Reference Type:

publication

Title:

Chromosome Cytology.

Author:

Moorhead, P.S. and P.C. Nowell

Year:

1964

Bibliographic source:

In Methods In Medical Research, H. N. Eisen, Ed. Yearbook Medical Publ., Inc., Chicago, p. 310.

Reference 4

Reference Type:

publication

Title:

Virus-cell interaction with tumor producing virus.

Author:

Vogt, M. and R. Dulbecco

Year:

1960

Bibliographic source:

Proc. Nat. Acad. Sci., 46, 365 - 370

Reference 5

Reference Type:

publication

Title:

Chromosome Praparations of Leukocytes Cultured from Humn Peripheral Blood

Author:

Moorhead, P.S:, Nowell, W.J., Mellman, W.J., Battips, D.M., Hunger, D.A.

Year:

1960

Bibliographic source:

Expt.Cell Res., 20: 613

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)

Principles of method if other than guideline:

Kidney cells from kidney tissue removed at autopsy were cultured by the trypsinization method of Fernandes (1958). Cultures were maintained in Eagle's medium as modified by Vogt and Dulbecco (1960). Cells were processed for spreading on slides by the method of Moorhead & Newell (1964). Slides were stained with Giemsa stain. Chromosomes were counted and morphological aberrations were examined from photographic negatives of up to 50 metaphase cells [ Lee, et al., (1976)].DEVIATIONS FROM OECD PROTOCOL #475. This study was carried out on rats that had been dosed in their diet for 5 and 13 weeks, respectively. The study also had no positive control. Two hundred cells were evaluated / rat for measuring cell ploidy. Kidney samples from fewer than five rats total were evaluated. The report does not specify whether the kidneys were from males, females, or both. Mitotic indices were not calculated.

GLP compliance:

not specified

Type of assay:

chromosome aberration assay

Test material

Test material information

Constituent 1

Reference substance name:

Glycerol trinitrate

EC Number:

200-240-8

EC Name:

Glycerol trinitrate

Cas Number:

55-63-0

Molecular formula:

C3H5N3O9 C3H5(NO3)3

IUPAC Name:

propane-1,2,3-triyl trinitrate

Details on test material:

- Name of test material (as cited in study report): TNG, trinitroglycerin
- Source: SDM No.17, Atlas Chemicals Division, ICI America Inc., Wilmington, Del.)
- Physical state. a 10% mixture on lactose

Test animals

Species:

rat

Strain:

other: CD® rats

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS- Source: Charles River Breeding Lab., Wilmington, Mass.- Fasting period before study: ad libitum- Housing: plastic cage with filter tops- Diet (e.g. ad libitum): ad libitum- Water (e.g. ad libitum): ad libitum- Acclimation period: 2 weeksENVIRONMENTAL CONDITIONS- Temperature (°C): 75+/-5°F- Humidity (%):50+/-10%- Air changes (per hr):10 air chenges per hour- Photoperiod (hrs dark / hrs light): 12-hour on and 12-hour off

Administration / exposure

Route of administration:

oral: feed

Vehicle:

- Vehicle(s)/solvent(s) used: lactose- Justification for choice of solvent/vehicle: TNG was desensitized in lactose.- Concentration of test material in vehicle: 10%

Details on exposure:

DIET PREPARATION- Rate of preparation of diet (frequency): weekly- Mixing appropriate amounts with (Type of food): standard rodent chow- Storage temperature of food: room temperature

Duration of treatment / exposure:

4 weeks or 13 weeks

Frequency of treatment:

24 hrs/day - The animal feeders were refilled after the 4th day and emptied completely after the 7th day before the freshly prepared TNG-feed mixture was used.

Post exposure period:

4 weeks

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:59 mg/kg/day Basis:nominal in dietmale average daily intake for first 5 weeks (from highest dose level), TNG intake was corrected for evaporation loss.

Doses / concentrations 2

Remarks:

Doses / Concentrations:229.5 mg/kg/dayBasis:nominal in dietmale average daily intake for weeks 6-13, incl. (from highest dose level). TNG intake was corrected for evaporation loss.

Doses / concentrations 3

Remarks:

Doses / Concentrations:59.3 mg/kg/dayBasis:nominal in dietfemale average daily intake for first 5 weeks (from highest dose level). TNG intake was corrected for evaporation loss.

Doses / concentrations 4

Remarks:

Doses / Concentrations:233.8 mg/kg/dayBasis:nominal in dietfemale average daily intake for weeks 6-13, incl. (from highest dose level). TNG intake was corrected for evaporation loss.

No. of animals per sex per dose:

16 females and 16 males per dose

Control animals:

yes, concurrent vehicle

Examinations

Tissues and cell types examined:

kidney cultures

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: Since the rat kidneys were from the highest dose group and the investigators concluded that there appeared to be no effect on the parameters measured, chromosome aberration analyses were not carried out on kidney cells from rats dosed at lower levels.KIDNEY CULTURESKidney tissue samples were removed at necropsy and cultured by the trypsinization method of Fernandes. All cultures were maintained in Eagle´s medium as modified by Vogt and Dulbecco.CHROMOSOME ANALYSISActively dividing kidney cultures and phytohemaglutin-stimulated lymphocytes were arrested in metaphase by short-term colchicine treatment. The cells were trypsinized, swollen in hypotonic solution and processed for spreading on glass slide by the method Moorhead and Nowell. Slides were stained with giemsa and scanned under low power optics. Cell polyploidy was estimated by examination of 200 cells. Slides showing minimum scattering of cells in metaphase were selected for analysis under oil immersion optics.

Evaluation criteria:

Chromosomes were counted and morphological aberration were examined from photographic negatives of up to 50 metaphase cells.

Results and discussion

Test results

Sex:

not specified

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

not specified

Any other information on results incl. tables

Rat kidney cells

Daily dose (mg/kg)	No.of rats	Chromosome frequency					Tetraploids per 100 cells
		40	41	42	43	44
WKS 1-5
0	4	4^1	4	42	1	0	0.59±0.19^2
Ave.59	3	3	1	44	1	0	0.65±0.33
WKS 6-13
0	4	4	4	42	1	0	0.59±0.19
Ave.229.5	4	4	2	43	1	0	0.75±0.25

¹mean,²mean±standard error

Dose (mg/kg)	No. of rats	Chromatid breaks&gaps/50 cells	Translocations/50 cells	Total aberrations/50 cells
WKS 1-5
0	4	1.5±0.3^1	0.5±0.6	2±0.3
Ave.59	3	2±0.6	1±0.1	2±1.3
WKS 6-13
0	4	1.5±0.3	0.5±0.3	2±0.3
Ave.233.8	4	1±0.7	0.5±0.3	1.5±0.9

¹mean

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negativeThe data are indicative, but incomplete by contemporary standards. There is no apparent suggestion of an effect, but the small number of control and test rats evaluated and the lack of information as to the sexes involved make statistical evaluation of the results questionable. The test animal standard errors usually were larger than those for the control animal physical aberration measurements, suggesting more scatter in the test animal measurements, but there does not appear to be any real suggestion of an effect.