3-chloropropene

Repeated dose toxicity: inhalation

Administrative data

Endpoint:

sub-chronic toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1977-09-27 - 1982-04-20

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: - non-GLP - generally guideline compliant

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1982

Report date:

1982

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 413 (Subchronic Inhalation Toxicity: 90-Day Study)

Deviations:

yes

Remarks:

: limited amount of clinical biochemistry and hematology

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

3-chloropropene

EC Number:

203-457-6

EC Name:

3-chloropropene

Cas Number:

107-05-1

Molecular formula:

C3H5Cl

IUPAC Name:

3-chloroprop-1-ene

Details on test material:

- Name of test material (as cited in study report): allyl chloride
- Substance type: chlorinated short-chain hydrocarbon
- Physical state: liquid
- Analytical purity: Charge I: 98.9, Charge II: 98.7
- Impurities (identity and concentrations): (wt %)
Charge I:
Allyl Chloride 98.9
2-Chloropropene 0.02
Isopropyl Chloride 0.42
1-5 Hexadiene 0.37
Normal propyl chloride 0.26
Acetonitrile 0.05

Charge II:
Allyl Chloride 98.7
2-Chloropropene 0.01
Isopropyl Chloride 0.47
1-5 Hexadiene 0.48
Normal propyl chloride 0.25
Acetonitrile 0.01

- Purity test date: not reported
- Lot/batch No.: Charge I: TB06037-1, Charge II: Lot Ho. TB06167-1
- Supplier: Water-washed allyl chloride, representative of productiongrade material, was obtained from The Dow Chemical Company, Freeport, Texas.
- Expiration date of the lot/batch: not reported
- Stability under test conditions: not reported, but expected to be stable
- Storage condition of test material: not reported

Test animals

Species:

rat

Strain:

Fischer 344

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories, Wilmington, Massachusetts, U.S.A.
- Age at study initiation: approximately 6 weeks of age at acclimatization and 8 weeks at exposure
- Weight at study initiation: males 204 - 205 g (average), females: 143 - 145 g (average) at 10 d prior to exposure
- Fasting period before study: no
- Housing: 3 or 4/cage,
- Diet (e.g. ad libitum): not specified, ad libitum
- Water (e.g. ad libitum): not specified, ad libitum
- Acclimation period: 2 wks


ENVIRONMENTAL CONDITIONS
not reported

Administration / exposure

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

other: unchanged (no vehicle)

Remarks on MMAD:

MMAD / GSD: not applicable, exposure by vapors of a volatile chlorinated short-chain hydrocarbon

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 14.5 m³, chambers with steel ceilings in the shape of a regular quadrangular pyramid and an epoxy resin coating on the walls and floors
- Method of holding animals in test chamber: exposed in home cages
- Source and rate of air: not reported
- Method of conditioning air: controlled for temperature and humidity
- System of generating vapor: metering a calculated amount of 3-chloropropene liquid into a heated (approximately 80 ° C) vaporization flask with a precision syringe pump and sweeping the vapor with filtered air (controlled for temperature and humidity) into the main chamber
- Temperature, humidity, pressure in air chamber: temp: 20.3 - 24 °C (averages), hum: 40.5 - 60.6 % (averages, pres: not reported
- Air flow rate: 2200-2900 liters per minute
- Air change rate: 9-12 air changes per hour
- Treatment of exhaust air: not reported


TEST ATMOSPHERE
- Brief description of analytical method used: The analytical concentration of allyl chloride vapor in each chamber was determined 5 or 6 times per day using a Miran IA-CVF infrared spectrophotometer.
- Samples taken from breathing zone: not reported

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

The analytical concentration of ally' chloride vapor in each chamber was determined 5 or 6 times per day using a Miran IA-CVF infrared spectrophotometer. A strip chart recorder monitored IR output. Instrument settings were as follows:
Wavelength: 10.8 µ
Pathlength: 18.75 m
Slit width: 1 mm
Standards were prepared by injecting a calculated amount of allyl chloride liquid into a 100 liter Saran bag. All standard curves were drawn using linear regression analysis (Texas Instruments SR-51A linear regression function). Analytical values for chamber concentrations of allyl chloride were extrapolated from the standard curves. A 100 ppm standard was run each exposure day during the 90-day study.

Duration of treatment / exposure:

6 h/d

Frequency of treatment:

5d/wk

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 50, 100 and 250 ppm
Basis:
nominal conc.

No. of animals per sex per dose:

25

Control animals:

yes, sham-exposed

Details on study design:

- Dose selection rationale: based on a 4 d study
- Rationale for animal assignment (if not random): random
- Rationale for selecting satellite groups: 10 animals per group, designated prior to start exposure for an interim sacrifice after 30 d
- Post-exposure recovery period in satellite groups:
- Section schedule rationale (if not random):

Positive control:

no

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily
- list of conducted observations not presented in detail


DETAILED CLINICAL OBSERVATIONS: No


BODY WEIGHT: Yes
- Time schedule for examinations: before the initiation of exposure, twice weekly for the first 30 days of the study, and once weekly for the duration of the study


FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No


FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No


WATER CONSUMPTION: No


OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: at final sacrifice
- Dose groups that were examined: all dose groups

HAEMATOLOGY: Yes
- Time schedule for collection of blood: approximately one week prior to the designated interim and terminal necropsies
- Anaesthetic used for blood collection: not reported
- Animals fasted: No
- How many animals: 10 rats/group at interim sacrifice + 10 rats/group at terminal sacrifice, collected from the tail veins of the rats
- Parameters checked in table 1 were examined.


CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at terminal and interim sacrifice
- Animals fasted: Yes
- How many animals: 10 rats/group at interim sacrifice + 10 rats/group at terminal sacrifice, from the severed cervical vessels
- Parameters checked in table 1 were examined.


URINALYSIS: Yes
- Time schedule for collection of urine: approximately one week prior to the designated interim and terminal necropsies
- Metabolism cages used for collection of urine: Not reported
- Animals fasted: No
- Parameters checked in table 1 were examined.


NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (see table 2)
HISTOPATHOLOGY: Yes (see table 2)

Statistics:

Hematology, clinical chemistry, urinary specific gravity, organ weight, and body weight data were evaluated using an analysis of variance and Dunnett's test (Steel, R. G. D. and Torrie, H. H., Principles and Procedures of Statistics, McGraw-Hill, New York (1960), pp. 101-105; 111-112.).
The level of significance chosen for all cases was p<0.05.

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

not examined

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY
- initially in the 90-day study, male and female rats at all exposure levels and controls showed conjunctival redness and palpebral closure suggestive of irritation. This observation was due to an outbreak of infectious sialodacryoadenitis noted in these rats during the first 2 weeks of the study. In the 250 ppm exposed rats there was an occasional in-life observation suggesting that the feces were less well formed.
- However, no consistent observations were noted either in rats or mice which were considered of toxicologic significance due to allyl chloride exposure
- two fatalities occured, one male of the control group and one female of the mid-dose group, both not regarded exposure related

BODY WEIGHT AND WEIGHT GAIN
- no statistically significant differences in body weights were detected

FOOD CONSUMPTION
- not analyzed

FOOD EFFICIENCY
- not analyzed

WATER CONSUMPTION
- not analyzed

OPHTHALMOSCOPIC EXAMINATION
- no effects detected

HAEMATOLOGY
- red blood cell parameters reduced in the 250 ppm group

CLINICAL CHEMISTRY
- AP levels were reduced in a generally dose dependent manner, becoming statistically significant in both high dose groups at both time points, but judged not to be of toxicological significance
- no other treatment related effects were found

URINALYSIS
- No biologically significant change was noted in the urinalyses parameters for male or female rats from the interim or terminal kill which indicated an exposure-related effect of toxicologic significance.

NEUROBEHAVIOUR
- not examined

ORGAN WEIGHTS
- at the terminal sacrifice after 90 d both livers and kidneys show a generally dose dependent rise in absolute and relative weights. At 100 ppm these changes are always statistically significant and > 10% as compared to the control animals, therefore this finding is regarded as adverse effect at this level.

GROSS PATHOLOGY
- though severall observations were made ( lungs, liver, kidneys and thymus) no general pattern or trend could be determined regarding different exposure levels and the two time points. Therefore a treatment related effect could not be determined.

HISTOPATHOLOGY: NON-NEOPLASTIC
- kidneys:
increased cytoplasmic granularity and eosinophilic staining of the cortical epithelial cells in the 100 and 250 ppm groups. In addition, a
greater number of the male and female rats in the 250 ppm group contained slightly more tubules in their kidneys with focal collapse and atrophy.
The changes were considered exposure-related.
- lung:
In the lungs of nearly all male and female rats in the 250 ppm group there were microscopic changes characterized as focal granulomatous inflammation, where the lesions contained a foreign plant or hairlike material within their center.
The microscopic change present in the lungs of these rats was not characteristic of a chemically induced pneumonitis, and therefore was not considered a direct result of 3-chloropropen exposure.

HISTOPATHOLOGY: NEOPLASTIC (if applicable)
- no effects detected

HISTORICAL CONTROL DATA (if applicable)
- not reported

Effect levels

1

Dose descriptor:

NOAEC

Effect level:

50 ppm

Sex:

male/female

Basis for effect level:

other: absence of any statistically or biologically significant effects

2

Dose descriptor:

LOAEC

Effect level:

100 ppm

Sex:

male/female

Basis for effect level:

other: slight increase in the cytoplasmic granularity and eosinophilic staining of the cortical epithelial cells in combination with increasing relative and absolute kidney weights.

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

- measured concentrations: 50 ppm: 50.5 ± 2.7 ppm; 100 ppm: 100.9 ± 2.6 ppm; 250 ppm: 250.4 ± 6.5 ppm

- Table 3: Average body weights and body weight gains during 89 days of treatment

Dose rate (ppm)	Body Weights (g)				Total Weight Gain
	day -9	day 3	day 25	day 89	g	% of control
Male
0	205.2 ± 9.5	217.1 ± 17.1	280.2 ± 15.1	361.3 ± 17.4	156.1	-
Low	205.7 ± 10.1	196.9 ± 9.8 *	277.6 ± 15.3	342.9 ± 17.6	137.2	87.9
Mid	204.2 ± 13.1	180.8 ± 10.4**	278.0 ± 15.3	358.7 ± 27.9	154.5	99.0
High	204.9 ± 10.9	219.1 ± 12.4	275.2 ± 15.9	348.2 ± 31.6	143.3	91.8
Female
0	144.0 ± 6.7	154.1 ± 7.3	175.4 ± 6.8	193.6 ± 7.8	49.6	-
Low	145.4 ± 6.3	148.2 ± 7.0 *	173.1 ± 6.6	193.1 ± 6.3	47.7	96.2
Mid	142.9 ± 4.3	125.9 ± 5.0 **	171.9 ± 6.9	194.5 ± 7.4	51.6	104.0
High	145.2 ± 4.1	150.6 ± 6.3	169.0 ± 5.4 *	181.7 ± 9.3 *, a	36.5	73.6

* Significantly different (p <0.05) from the control.

** weight difference due to problems with automatic watering system

a: no significant weight effect between d 45 and day 82 after first exposure, weight loss between day 82 and 89

- Table 4: organ and body weights for male Fischer 344 rats exposed to allyl chloride vapor 6 hours per day, 5 days per week

Days on Test	Vapor Conc. (ppm)	Number of Rats /Group	Sex	Fasted Body Weight (g)	Organ Weights (in g or g/100 g of body weight)
					Liver		Kidney		Brain		Heart		Spleen		Thymus		Testes
					g	g/100	__g__	g/100	g	g/100	__g__	g/100	__g__	g/100	__g__	g/100	__g__	g/100
29	0	(10)	M	253	6.80	2.69	1.93	0.76	1.80	0.71	0.79	0.31	0.53	0.21	0.29	0.11	2.96	1.18
				± 16	± 0.55	± 0.18	± 0.10	± 0.03	± 0.03	± 0.04	± 0.05	± 0.01	± 0.06	± 0.02	± 0.03	± 0.01	± 0.11	± 0.07
29	50	(10)	M	254	6.87	2.71	2.06	0.81*	1.79	0.71	0.81	0.32	0.50	0.20	0.26	0.10	2.98	1.18
				± 14	± 0.45	± 0.07	± 0.12	± 0.03	± 0.03	± 0.03	± 0.04	± 0.01	± 0.04	± 0.02	± 0.03	± 0.01	± 0.10	± 0.07
29	100	(10)	M	258	7.39*	2.86*	2.18*	0.85*	1.80	0.70	0.82	0.32	0.49	0.19	0.27	0.10	3.01	1.17
				± 10	± 0.48	± 0.10	± 0.11	± 0.03	0.02	± 0.02	± 0.04	0.01	± 0.06	± 0.02	± 0.04	± 0.01	± 0.08	± 0.04
29	250	(10)	M	254	7.36	2.89*	2.09*	0.82*	1.76	0.69	0.79	0.31	0.52	0.21	0.30	0.12	2.99	1.18
				± 15	± 0.62	± 0.10	± 0.02	± 0.05	± 0.05	± 0.03	± 0.05	± 0.01	± 0.07	± 0.03	± 0.03	± 0.01	± 0.12	± 0.05
95	0	(14)	M	342	8.58	2.51	2.36	0.69	1.94	0.57	0.95	0.28	0.60	0.18	0.24	0.07	3.20	0.94
				± 16	± 0.71	± 0.14	± 0.17	± 0.03	± 0.08	± 0.02	± 0.08	± 0.02	± 0.08	± 0.02	± 0.06	± 0.02	± 0.14	± 0.04
95	50	(13)	M	316*	8.50	2.69*	2.46	0.78*	2.01*	0.64*	0.95	0.30*	0.59	0.19	0.21	0.07	3.17	1.01*
				± 20	± 0.67	± 0.12	± 0.18	± 0.03	± 0.04	± 0.04	± 0.05	± 0.01	± 0.07	± 0.02	± 0.04	± 0.01	± 0.24	± 0.08
95	100	(13)	M	339	9.51*	2.81*	2.67*	0.79*	2.00	0.59	1.01	0.30*	0.61	0.18	0.24	0.07	3.24	0.96
				± 24	± 0.67	± 0.11	± 0.02	± 0.04	± 0.05	± 0.04	± 0.06	± 0.01	± 0.08	± 0.02	± 0.06	± 0.02	± 0.12	± 0.05
95	250	(13)	M	322	9.54*	2.95*	2.64*	0.82*	1.92	0.60	0.96	0.30*	0.60	0.19	0.19	0.06	3.20	1.00*
				± 29	± 1.17	± 0.14	± 0.23	± 0.03	± 0.06	± 0.05	± 0.07	± 0.01	± 0.06	± 0.01	± 0.06	± 0.02	± 0.21	± 0.06

All data listed au mean±S.D.

* Statistically significant deviation from control mean using Dunnett’s Test, p<0.05.

- Table 5: organ and body weights for female Fischer 344 rats exposed to 3-chloropropene vapor 6 hours per day, 5 days per week

Days on Test	Vapor Conc. (ppm)	Number of Rats /Group	Sex	Fasted Body Weight (g)	Organ Weights (g and g/100 g body weight)
					liver		kidney		brain		heart		spleen		thymus
					g	g/100	g	g/100	g	g/100	g	g/100	g	g/100	g	g/100
29	0	(10)	F	164	4.15	2.53	1.30	0.79	1.74	1.06	0.55	0.34	0.41	0.25	0.23	0.14
				± 17	± 0.28	± 0.13	± 0.06	± 0.04	± 0.04	± 0.05	± 0.03	± 0.01	± 0.05	± 0.02	± 0.03	± 0.02
29	50	(10)	F	160	4.30	2.68	1.33	0.83	1.70a	1.07	0.57	0.35	0.41	0.26	0.23	0.14
				± 18	± 0.43	± 0.20	± 0.10	± 0.06	± 0.03	± 0.04	± 0.03	± 0.01	± 0.04	± 0.02	± 0.03	± 0.01
29	100	(10)	F	161	4.27	2.65	1.36	0.84	1.70°	1.06	0.57	0.35	0.39	0.24	0.22	0.14
				± 6	± 0.17	± 0.07	± 0.05	± 0.03	± 0.02	± 0.03	± 0.03	± 0.01	± 0.02	± 0.01	± 0.03	± 0.01
29	250	(10)	F	151a	4.1 4	2.74°	1.31	0.87°	1.65a	1.09	0.56	0.37a	0.39	0.26	0.21	0.14
				± 6	± 0.23	± 0.12	± 0.02	± 0.03	± 0.03	± 0.03	± 0.02	± 0.02	± 0.05	± 0.03	± 0.02	± 0.01
95	0	(15)	F	179	4.52	2.53	1.39	0.78	1.86	1.04	0.61	0.34	0.45	0.25	0.21	0.12
				± 7	± 0.21	± 0.11	± 0.07	± 0.03	± 0.06	± 0.05	± 0.04	± 0.03	± 0.05	± 0.03	± 0.04	± 0.02
95	50	(15)	F	182	4.89°	2.69a	1.53a	0.85°	1.89	1.04	0.66a	0.36	0.44	0.24	0.21	0.12
				± 7	± 0.34	± 0.13	± 0.08	± 0.05	± 0.03	± 0.04	± 0.04	± 0.02	± 0.03	± 0.02	± 0.03	± 0.01
95	100	(15)	F	178	4.83°	2.71°	1.52a	0.86a	1.84	1.03	0.65	0.36	0.45	0.25	0.21	0.12
				16	± 0.1.7	± 0.11	± 0.07	± 0.05	± 0.03	± 0.07	± 0.03	± 0.02	± 0.02	± 0.01	± 0.04	± 0.02
95	250	(15)	F	168°	4.808	2.86°	1.48a	0.89a	1.79a	1.07	0.64	0.38	0.46	0.28a	0.20	0.12
				± 10	± 0.28	± 0.15	± 0.08	± 0.05	± 0.04	± 0.07	± 0.08	± 0.07	± 0.03	± 0.01	± 0.04	± 0.02

All data listed au mean±S.D.

* Statistically significant deviation from control mean usingDunnett’s Test, p<0.05.

Applicant's summary and conclusion

Executive summary:

In the present study (Quast 1982) Fischer 344 rats of both sexes were treated with 3 -chloropropene repeatedly via the inhalation route for 6 hours, ten animals per sex and dosage, 90 d, 5d/wk. Treatment concentrations were 0, 50, 100 and 250 ppm. Based on the presented results a NOAEC of 50 ppm and correspondingly a LOAEC of 100 ppm could be derived for the toxicity after repeated, subchronic exposure to 3 -chloropropene via the inhalative route.

Animals observed daily for clinical signs and were weighted at days -9 and -2 prior to the start of exposure and on day 2 post exposure and then twice weekly for the first 30 days of the study, and once weekly for the duration of the study.

Ten of 25 animals per dose group were designated for an interim sacrifice at day 30 after start of exposure. Of all animals parameters in hematology, clinical chemistry and urinalysis were determined and all were subjected to gross necroscopy and histologic samples were prepared and analyzed microscopically.

No clinical signs indicative of toxic effects of 3-chloropropene were reported. Two not exposure related fatalities occured. Body weights were not affected in any dose group. In the high dose groups the red blood cell parameters were slightly reduced which was the only finding in hematology. Clinical chemistry parameters were unremarkable except for reduced AP levels (in a generally dose dependent manner) becoming statistically significant in both high dose groups at both time points, but judged not to be of toxicological significance. No biologically significant exposure related change was noted in the urinalyses parameters for male or female rats from the interim or terminal kill. At the terminal sacrifice after 90 d both livers and kidneys show a generally dose dependent rise in absolute and relative weights. In the kidneys at 100 ppm these changes are always statistically significant and > 10% as compared to the control animals, therefore this finding is regarded as an adverse effect at this level. Though several observations were made ( lungs, liver, kidneys and thymus) at the gross necroscopy no general pattern or trend could be determined regarding different exposure levels and the two time points. Therefore a treatment related effect could not be determined.

In parallel to the kidney weight effects an increased cytoplasmic granularity and eosinophilic staining of the cortical epithelial cells was found in the 100 and 250 ppm groups. In addition, a greater number of the male and female rats in the 250 ppm group contained slightly more tubules in their kidneys with focal collapse and atrophy. The changes were considered exposure-related.

In addition in the lungs of nearly all male and female rats in the 250 ppm group there were microscopic changes characterized as focal granulomatous inflammation, where the lesions contained a foreign plant or hairlike material within their center.

The microscopic change present in the lungs of these rats was not characteristic of a chemically induced pneumonitis, and therefore was not considered a direct result of 3-chloropropen exposure.

In conclusion a slight effect on the weight and the histopathology of the kidney can be observed after the repeated exposure to 3 -chloropropene via the inhalative route at 100 ppm and higher concentrations. At higher concentrations also liver weight effects can be detected. The kidneys are regarded to be the primary target organ.