Styrene

Specific investigations: other studies

Administrative data

Endpoint:

specific investigations: other studies

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study.

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference 1

Reference Type:

publication

Title:

Ring-oxidized metabolites of styrene contribute to styrene-induced Clara-cell toxicity in mice.

Author:

Cruzan G, Carlson GP, Turner M, Mellert W.

Year:

2005

Bibliographic source:

J Toxicol Environ Health, Part A 68(3): 229-237

Reference 2

Reference Type:

review article or handbook

Title:

European risk assessment report, Styrene CAS No. 100-42-5, EINECS No. 202-851-5, Draft for submission to SCHER, November 2007.

Author:

European Union

Year:

2007

Bibliographic source:

Styrene CAS No. 100-42-5, EINECS No. 202-851-5, Draft for submission to SCHER, November 2007

Materials and methods

Principles of method if other than guideline:

In a study conducted to investigate the role of the ring-oxidized metabolite of styrene, 4-VP, in mouse lung toxicity, groups of 4-5 male CD-1 mice received a single ip injection of 0 or 100 mg/kg 4-VP.

GLP compliance:

yes

Type of method:

in vivo

Endpoint addressed:

not applicable

Test material

Test material information

Constituent 1

Reference substance name:

Styrene

EC Number:

202-851-5

EC Name:

Styrene

Cas Number:

100-42-5

Molecular formula:

C8H8

IUPAC Name:

ethenylbenzene

Specific details on test material used for the study:

The substance tested is a metabolite of styrene.

Test animals

Species:

mouse

Strain:

CD-1

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River, Wilmington, MA
- Weight at study initiation: 22-24 g
- Housing: in air conditioned rooms
- Diet (e.g. ad libitum): standard rodent diet (Purina 5001, Purina Milss, St. Louis, MO) or pellets (Provimi KLIBA SA, Kaiseraugst, Switzerland)
- Water (e.g. ad libitum): from water bottles


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24 °C
- Humidity (%): 30-70%
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

intraperitoneal

Vehicle:

physiological saline

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

single injection

Frequency of treatment:

once

Post exposure period:

12, 24, 48 and 72 h, 4 and 6 d

Doses / concentrations

Remarks:

Doses / Concentrations:
100 mg/kg bodyweight
Basis:
nominal conc.

No. of animals per sex per dose:

4-5

Control animals:

yes

Details on study design:

Groups of 4 to 5 male CD-1 control mice or mice administered 100 mg/kg 4-vinylphenol ip (weighing 22-24 g) were sacrificed after 12, 24, 48 or 72 h or 4 or 6 d. Mice were anesthetized with diethyl ether; the trachea was exposed, where a small nick was made and an oral feeding needle inserted and tied in place. The lungs were lavaged twice with 0.8 ml of room-temperature lavage fluid (total 1.6 ml); recovery was about 85%. The fluid consisted of NaCl (145 mM), KCl (5 mm), NaH 2PO 4 (1.9 mm), Na2HPO4 (9.4 mM), and glucose (5.5 mm) at a pH of 7.4.

The difference from control was evaluated using Student's test (Instat by GraphPad Software, San Diego, CA) with a significance value of p < 0.05. Serum sorbitol dehydrogenase was evaluated by the Kruskal-Wallis nonparametric test (Siegel, 1956) with a significance value of p < 0.05.

Examinations

Examinations:

The number of cells in 100 µl bronchoalveolar lavage fluid (BALF) was counted using a hemocytometer. The remaining BALF from each mouse was centrifuged at low speed, the amount of protein was determined using the bicinchoninic method (Redinbaugh & Turley, 1986), and lactate dehydrogenase (LDH) activity was measured by the spectrophotometric method of Vassault (1983).

Results and discussion

Details on results:

4-Vinylphenol administration resulted in lung damage as measured by increased lactate dehydrogenase (LDH) activity (approx. 2-fold compared to controls), and cells in bronchoalveolar lavage fluid (BALF) 12 h after a single 100-mg/kg ip administration to CD-1 mice. LDH activity remained elevated until d 4; the number of cells remained elevated until d 6 (approx. 7- to 20-fold compared to controls). Protein was significantly elevated only on d3.

Any other information on results incl. tables

	Lactate dehydrogenase (a)		Cells (b)		Protein (c)
Time after application	Control	4-VP	Control	4-VP	Control	4-VP
12 h	125.6 ± 26.3	200.2 ± 12.8 (d)	109 ± 37	795 ± 116 (d)	524 ± 59	452 ± 67
24 h	92.5 ± 23.1	254.2 ± 26.0 (d)	40 ± 14	1104 ± 89 (d)	408 ± 48	454 ± 52
48 h	97.5 ± 21.0	210.1 ± 25.7 (d)	44 ± 13	753 ± 230 (d)	360 ± 40	486 ± 63
72 h	73.0 ± 14.1	156.6 ± 14.7 (d)	76 ± 14	1415 ± 41 (d)	360 ± 28	598 ± 68 (d)
4 d	89.2 ± 16.9	86.1 ± 4.5	67 ± 16	345 ± 200 (d)	368 ± 25	478 ± 51
6 d	77.8 ± 9.1	104.5 ± 11.8	76 ± 21	51 ± 15	442 ± 10	520 ± 33

(a)   µmol/min/L of bronchoalveolar lavage fluid

(b)  cells/µL bronchoalveolar lavage fluid

(c)   µg protein/ µL bronchoalveolar lavage fluid

(d)  significantly different from control p<0.05