3-chloropropene

Basic toxicokinetics

Administrative data

Endpoint:

basic toxicokinetics in vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: - scientifically sound study - non-GLP, no guideline followed, but well documented

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1982

Report date:

1982

Materials and methods

Objective of study:

toxicokinetics

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

- F344 rats were treated with 3-chloropropene by 3 different routes in 3 different experiments
- Experiment A: single oral gavage of labelled test item at 1 and 100 mg/Kg, air trapping; urine and feces sampling (metabolic cages) at blood samples at 1, 2, 3, 4, 6, 8, 16, 24, 32, 40 and 48 h post dosing from indwelling jugular cannulas; determination of radioactivity in carcass and samples; sacrifice of animals 48 h post dosing and histopathologic analysis of liver and kidney tissue samples; bile cannulation of 2 rats; urinary metabolite profile determination from 2 animals
- Experiment B: single oral gavage or single intravenous injection of non-labelled test item at 100 mg/Kg, blood sampling at 5, 10, 15, 30, 60, 120, and 180 minutes after dosing indwelling jugular cannulas.
-Experiment C: Fasted rats exposed via inhalation at 10, 100, 1000 and 2000 ppm of unlabeled test item for 6 h in head-only exposure chambers; blood sampling at 0.25, 0.5, 1, 2, 4 and 6 h during exposure and at different time points thereafter; histopathologic analysis of liver and kidney tissue samples

GLP compliance:

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
- Physical state: not reported
- Analytical purity:
non-labeled-1: 99 % (mole percent, NMR by J.P. Heeschen of the Analytical Laboratories); used in Experiment A
non-labeled-2: 97.7 %, GC-analysed (E. A. Hermann, Toxicology Laboratories, The Dow Chemical Company, Midland, MI); used in Excperiments B and C;
labeled: 72± 12 volume percent allyl chloride and 15 ± 3 volume percent diethyl ether; used in Experiment A
- Impurities (identity and concentrations): see above, no further details given
- Purity test date: not reported
- Lot/batch No.:
non-labeld-1: #SI 3239457 from Inhalation Group, Toxicology Laboratories, The Dow Chemical Company, Midland, MI
non-labeled-2: lot :4-TB 06308-3 from The Dow Chemical Company (Freeport, TX).
labeled: lot #70202 from Pathfinder Laboratories (St. Louis, MO)
- Specific activity (if radiolabelling): ; 1.2 mCi/mmol
- Locations of the label (if radiolabelling): termed as (14C)-1,3-allyl chloride, but the main site of labeling could not be determined ("The degree of incorporation at any one site was within ± 5 percent (absolute) of the degree of incorporation at any other site.")
- Expiration date of radiochemical substance (if radiolabelling): not reported
- Stability under test conditions: not reported, but expected to be stable
- Storage condition of test material: not reported

Radiolabelling:

yes

Remarks:

all C-atoms evenly labeled

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
- Age at study initiation: not reported
- Weight at study initiation:
Experiment A (main experiment): males: 160 - 181 g, females: 124 - 153 g; Experiment A (bile cannulated animals): males: 224 - 237 g; Experiment A (metabolite profile) not reported
Experiment B (gavage): males: 216 - 248 g; no information on the animals for the iv application
Experiment C: males, blood sampling: 202 - 250 g; males, NPSH analysis: 233 - 283 g
- Fasting period before study: yes, overnight, except for the animals in Experiment C, which were used for NPSH analysis
- Housing: individually
- Individual metabolism cages: yes
- Diet (e.g. ad libitum): not reported
- Water (e.g. ad libitum): not reported
- Acclimation period: yes, but duration not detailed

ENVIRONMENTAL CONDITIONS
- no details reported

Administration / exposure

Route of administration:

other: gavage, intravenous injection and inhalative (head only)

Vehicle:

corn oil

Remarks:

for oral and injection experiments, unchanged for inhalation experiment

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Justification for use and choice of vehicle (if other than water): not reported, but cornoil is a standard vehicle for volatile solvents
- Concentration in vehicle:
Experiment A: 1 or 100 mg/mL (in both cases 5 µCi/kg of 14C-labeled compound)
Experiment B: gavage 33.3 mg/mL; iv injection: NO vehicle used
- Amount of vehicle (if gavage):
Experiment A: 1 mL/Kg bw
Experiment B: 3 mL/Kg bw


TYPE OF INHALATION EXPOSURE: head only

GENERATION OF TEST ATMOSPHERE / CHAMPER DESCRIPTION
- Exposure apparatus:
blood sampling: head-only inhalation chamber (Hefner, R.E., Jr., Watanabe, P.G. and Gehring, P.J. (1975)- Preliminary studies of the fate of inhaled vinyl chloride monomer in rats. - Ann, N.Y. Acad. Scin. 246: 135-148.) under dynamic airflow conditions.
NPSH analysis: 112 L glass and stainless steel chamber within a fume hood
- Method of holding animals in test chamber: not reported
- Source and rate of air: 15 to 23 L/Min
- Method of conditioning air: not reported
- System of generating test atmosphere:
100, 1000 and 2000 ppm exposures: metering a calculated amount of 3-chloropropene liquid into a J-tube assembly with a total chamber airflow ranging from approximately 15 to 23 L/min supplied through the J-tube assembly (Miller, R.R., Letts, R.L., Potts, W.J. and McKenna, M.J. (.1980) Improved methodology for generating controlled test atmospheres Am. Ind. Hyg. Assoc. J. 41: 844-846).
10 ppm: pumping allyl chloride vapor from a 100 L SARAN* bag (containing 1250 ppm) at a rate of 200 ml/min into the J-tube assembly with a total chamber airflow of approximately 25 L/min.
- Method of particle size determination: not applicable, vapor of a volatile solvent
- Treatment of exhaust air: not reported
- Analytic: infrared spectrometer (Wilks Miran LA; Foxboro Analytical, So. Norwalk, CT)

Duration and frequency of treatment / exposure:

Experiment A and B, gavage and iv injection: single treatment
Experiment C, inhalation exposure: single treatment for 6 h

Doses / concentrations

Remarks:

Doses / Concentrations:
Experiment A (main experiment): 1 and 100 mg/Kg bw
Experiment A (bile cannulated animals): 100 mg/Kg bw
Experiment A (urinary metabolites profiling): 100 mg/Kg bw
Experiment B: 100 mg/Kg bw
Experiment C: 10, 100, 1000, 2000 mg/m³

No. of animals per sex per dose / concentration:

Experiment A: 5 males and 5 females
Experiment A (bile cannulated animals): 2 males
Experiment A (urinary metabolites profiling): 2 males
Experiment B: gavage: 4 males, iv injection: 3 males
Experiment C: blood sampling: 3 males and 3 females (except for 200 ppm: total of 4 animals); NPSH analysis: 6 rats in total for each exposure limit

Control animals:

other: yes, either treated with vehicle (gavage) or sham exposed (inhalation)

Positive control reference chemical:

no

Details on study design:

- Dose selection rationale: not reported

Details on dosing and sampling:

PHARMACOKINETIC STUDY (Absorption, distribution, excretion)
- Tissues and body fluids sampled:
Experiment A (main experiment):
- air trapping (5M ethanolamine in 2- methoxyethanol at 8 hour intervals after dosing for 24 hours);
- urine sampling (8 h intervals, maintained on dry ice); feces sampling (24 h periods, at room temperature);
- skin samples (frozen, pulverized and combusted)
- blood samples at 1, 2, 3, 4, 6, 8, 16, 24, 32, 40 and 48 h post dosing from indwelling jugular cannulas; erythrocyte count from samples at 4, 8, 24 and 48 h post dosing
- determination of radioactivity in carcass (combustion and CO2 trapping) and samples by liquid scintilation counting;
- sacrifice of animals 48 h post dosing and histopathologic analysis of liver and kidney tissue samples.
Experiment A (bile cannulation):
- bile and urine collection from 1.5 h post treatment to 24 h post treatment
Experiment A (urinary metabolite profiling):
- urine from 2 males collected (duration not noted), separated via liquid cation exchange chromatography and analysed via szintilation counting; comparison with a similiarly derived chromatogrm for epichlorhydrin (Langvardt, P.W. (1978) Preliminary observations on the urinary metabolites of apichlorchydrin and allyl chloride. Draft Report to CMA.)
Experiment B:
- blood sampling at 5, 10, 15, 30, 60, 120, and 180 minutes after dosing indwelling jugular cannulas; urine collection at 8, 12 and 24 h after dosing
Experiment C:
Fasted rats exposed via inhalation at 10, 100, 1000 and 2000 ppm of unlabeled test item for 6 h in head-only exposure chambers; blood sampling at 0.25, 0.5, 1, 2, 4 and 6 h during exposure and at different time points thereafter; histopathologic analysis of liver and kidney tissue samples; NPSH
was analyzed by D.A. Dittenber, (Toxicology Laboratories, The Dow Chemical Company, Midland, MI) using the method of Sedlak and Lindsay, 1968 as modified for use on the Centrifichem 400 (Union Carbide Corporation, Rye NY).Sedlak, J. and Lindsay, R.H. (1968) Estimation of Total, protein bound, and non-protein sulfhydryl groups in tissue with Ellman's reagent. Anal. Biochem. 25: 192-205.)
- Time and frequency of sampling: see above
- Method type(s) for identification: see above

Results and discussion

Main ADME results

Main ADME results 1

Type:

absorption

Results:

Absorption is dose dependent. At 1 mg/Kg bw via gavage at least 75 % of the substance is absorbed, at 100 mg/Kg bw via gavage at least 63 % are absorbed. For inhalative absorption see below.

Main ADME results 2

Type:

metabolism

Results:

3-chloropropene is extensively metabolised: at 1 mg/Kg bw (gavage) about 36 % of the radioactivity is excreted as CO2. At 100 mg/Kg bw the capacity of metabolism seems to be reached as only ca. 5.5 % are exhaled as CO2 while 18 % are exhaled unchanged.

Main ADME results 3

Type:

excretion

Results:

At 1 mg/Kg (gavage) bw ca. 35 % of the substance are excreted each via urine and exhalation and 3.5 % via feces (but see remark). At 100 mg/Kg bw (gavage) ca. 37 % are excreeted via urine, 23 % via exhalation and 1.5 % via feces (but see remark)

Toxicokinetic / pharmacokinetic studies

Details on absorption:

ORAL ABSORPTION
- The total oral absorption cannot be determined precisely as at 1 mg/Kg only 80.6 % of the applied radioactivity was recovered and at 100 mg/Kg only 64.8 % were recovered. Kaye (1972) had shown that only metabolites are present in the urine. So for 1 mg/Kg bw at least 70 % absorption can be assumed (35.7 urine + 34.2 % CO2) while for 100 mg/Kg bw at least 42.2 % absorption can be assumed.
The authors state that the low amount of radioctivity in feces might be the result of enterohepatic cycling. Another possible explanation might be that significant amount of 3-chloropropene or its metabolised evaporated from feces as they were sampled for 24 h at room temperature and at least 3-chloropropeneme has a vapor pressure that is higher than the one of acetone (see Table 1 for details)
- 3-chloropropene is absorbed readilyat low doses, but the velocity of absorption is dose dependent:
at 1 mg/Kg bw the blood peak concentration is at 1 h after treatment (Cmax = 0.4 ppm), while at 100 mg/Kg bw the peak was found after 4 h (Cmax =28.4 ppm) indiocative of a delayed absorption
INHALATIVE ABSORPTION
3-chloropropene is readily taken up via the inhalative route, but the even after 6 h the animals in any of the dose groups have reached a steady state. Absorption rates for inhalation are 0.89, 9.83, 63.9 and 67.5 μg/min for 10, 100, 1000 and 2000 ppm for 6 h respectively

COMPARISON
A comparison of the AUC's for the blood concentration time curves following oral, intravenous and inhalation exposure gives a relative index of the bioavailability of the parent compound via each route:
100 mg/kg oral dose would be approximately equivalent to to a six hour-150 ppm inhalation exposure
100 mg/kg intravenous dose would be approximately equivalent to a six hour-85 ppm exposure

Details on distribution in tissues:

- detailed data on distribution is not available. Given that 3-chloropropene is apolar, has a low molecular weight of 76,53 g/mol and has a Log Pow of 2.1 it can be expected that the substance is readily distributed troughout the body via the blood circulation.

Details on excretion:

Excretion is described to occur maily via urine and exhalation:
1 mg/Kg bw (gavage): 35.7 % via urine, 34.2 % via exhalation as CO2, 1.5 % via exhalation unmetabolised, 3.5 % via feces.
100 mg/Kg bw (gavage): 36.7 % via urine, 5.4 % via exhalation as CO2, 18.1 % via exhalation unmetabolised, 1.5 % via feces.
- In an additional experiment 12 % of the radioactivity was excreeted via the bile (after treatment with 100 mg/Kg bw orally). So the low value found in feces might either be an indicator for enterohepatic cycling or as described above higher amounts might truely be excreeted via feces but might have been lost due to the high vapor pressure of 3-chloropropene and the sampling duration of 24 h at room temperature.
- Excretion via urine is relatively fast wit a peak within the first 8 h after treatment for 1 mg/Kg bw and a peak between 8 and 16 h for 100 mg/Kg bw probably due to the delayed uptake in the latter case.

Toxicokinetic parameters

Test no.:

#1

Toxicokinetic parameters:

other: for toxicokinetic parameters see table 3

Metabolite characterisation studies

Metabolites identified:

no

Details on metabolites:

- Metabolites have not been identified but 3-chloropropene exposure via the inhalative route lead to a strong depletion of non-protein sulfhydryls which is indicative of phase-1 metabolism reactions where the substance or its metabolites react with GSH.

Any other information on results incl. tables

- Table 1: percentage of a single oral dose of 14C-allyl chloride excreted in urine of six male and five female rats during successive time intervals*

dose (mg/Kg)	sex	N	% of 14C recovered 48 hours after dosing
			urine	CO2	char-coal	feces	carcass	plasma	rbc's	skin	total
1	F	3	36.6 ±	37.7 ±	1.3 ±	4.6 ±	3.7 ±	1.2 ±	1.1 ±	-	86.3±
			5.3	3.7	0.1	0.2	0.6	0.2	0.9	-	8.5
1	M	3	34.7 ±	30.7 ±	1.7 ±	2.3 ±	3.7 ±	0.8 ±	0.02 ±	1.06±	74.9±
			1.8	8.7	1.3	0.2	0.3	0.1	0.06	0.2	6.8
1	M + F	6	35.7 ±	34.2 ±	1 .5 ±	3.5 ±	3.7 ±	1.0 ±	0.6 ±	-	80.6±
			3.7	7.1	0.9	1.3	0.5	0.3	0.8	-	9.3
100	F	2	32.8 ±	5.8 ±	17.2 ±	2.3 ±	1.5 ±	0.6 ±	0.8 ±	-	61.0±
			3.4	2.0	1.3	0.0	0.1	0.3	0.2	-	0.8
100	M	3	39.4 ±	5.1 ±	18.7 ±	1.0 ±	1.8 ±	0.5 ±	0.03 ±	0.7 ±	67.21
			5.9	1.5	6.2	0.3	0.2	0.1	0.01	0.3	1.4
100	M + F	5	36.7 ±	5.4 ±	18.1 ±	1.5 ±	1.7 ±	0.6 ±	0.3 ±	-	64.8:1
			5.8	1.5	4.5	0.7	0.2	0.2	0.5	-	3.6

*Means + S.D. for the number of animals listed above.

- Skin samples from these animals were not analyzed.

- Table 2: percentage of a single oral dose of 14C-allyl chloride excreted in urine of six male and five female rats during successive time intervals*

Excretion Interval (hr)	mg/kg bw	100 mg/kg bw
0-8	22.7±3.7	14.6±2.0
8-16	9.4±3.3	19.0±2.5
16-24	1.7±0.4	1.6±0.4
24-32	0.8±0.2	0.6±0.1
32-40	0.6±0.1	0.5±0.1
40-48	0.1:70.1	0.4±0.1.
Total	35.7±1.5	36.7±2.6

*Mean ± S.E.M. for five or six rats.

- Toxicokinetic parameters:

-half-life of 3 -chloropropene in blood after 100 mg/Kg via gavage: t1/2 = 2.58 h

-half-life of 3 -chloropropene in blood after 100 mg/Kg via iv injection: t1/2 = 23.5 min

-half-life of 3 -chloropropene in blood after inhalation of 100 ppm for 6 h: t1/2 = 16.7 min (other doses see below in table 3)

-the metabolism of 3 -chloropropene is shown by the fact that at 13 h after 100 mg/Kg via gavage the blood level of 3 -chloropropene is below 100 ppb, while the concentration of metabolites (measured by radioactivity) is > 1000 ppb even at 48 h after treatment.

-urinary metabolite profiles differ significantly between 3 -chloropropene and epichlorhydrin which suggests that epichlorhydrin is not the primary metabolite of 3 -chloropropene

- Table 3: Pharmacokinetic parameter estimates for different doses of inhaled allyl chloride (ac) in the rat

Chamber AC (ppm)	Absorption constant (µg/min)	End exposure Blood AC (ppm)	Alpha t1/2 (min)	Beta t1/2 (min)
10	0.89	0.06	3.2	25.0
100	9.83	1.07	2.5	16.7
1000	63.9	21.2	10.7	34.1
2000	67.5	47.7	33.0	108.1

- Table 4: Liver, kidney, lung and blood non-protein sulfhydryls (NPHS) after exposure to allyl chloride vapor for six hours

TREATMENT (PPM)		NPSH(a) (nmol/mg wet weight tissue or nmol/ml blood)
	LIVER	KIDNEY	LUNG	BLOOD
0	7.08 ±	4.15 ±	2.09 ±	1099.2 ±
	0.12	0.04	0.04	49.8
10	7.29 ±	4.27 ±	2.04 ±	1130.8 ±
	0.13	0.09	0.01	28.6
0	7.23 ±	4.37 ±	1.90 ±	1066.7 ±
	0.08	0.04	0.02	26.4
100	5.66 ±	4.15 ±	1.76 ±	1146.7 ±
	0.09*	0.06*	0.04*	31.0
0	7.34 ±	4.24 ±	1.92 ±	1045.0 ±
	0.18	0.07	0.03	60.6
1000	0.97 ±	0.94 ±	0.57 ±	880.0 ±
	0.01*	0.03*	0.02*	139.9
0	6.89 ±	4.04 ±	1.95 ±	606.6 ±
	0.17	0.21	0.02	24.9
2000	0.46 ±	0.37 ±	0.57 ±	446.7 ±
	0.06*	0.03*	0.02*	50.4*

a - Mean ± S.E.M. for five or six rats

* - Statistically significant at the p<0.05 level using Ounnett's test or Wilcoxon Rank-Sum Test.

- Gross necroscopy and Histopathology :

animals sacrificed after 48 h of recovery:

- A single oral or intravenous administration of 100 mg/kg dose of 3 -chloropropene to rats did not appear to result in any grossly observable changes in tissue morphology within 72 hours after dosing.

- A single inhalation exposure to 10, 100 or 1000 ppm 3 -chloropropene vapor for six hours did not appear to result in any grossly observable changes in tissue morphology within 48 hours after exposure.

- A single inhalation exposure to 2000 ppm 3 -chloropropene vapor produced mortality in all four rats within 8 hours following exposure. Externally these rats showed discolored hairs in the perineal region. Internally, the gastro-intestinal tract was empty and the cecum contained fluid. The livers showed a slight focal discoloration in the left lobe which was considered due to congestion and post-mortem change. All other viscera in these rats were also congested due to their spontaneous death.

- Light microscopic changes indicative of a treatment-related effect were not observed in the liver or kidneys of rats in the 10 or 100 ppm exposure groups.

- In the 1000 ppm exposure group of rats the liver was unaffected; however, the kidneys in all of these rats showed treatment-related changes. The microscopic findings in the kidneys were characterized by multifocal subacute degenerative and regenerative changes of the proximal convoluted tubules in the cortex.

animals sacrificed directly after treatment:

- Light microscopic examination of the left lobe of the liver and one kidney was performed on each control rat and each of the rats exposed to 10, 100 or 1000 ppm 3 -chloropropene for 6 hours and killed immediately thereafter. There were no recognizable microscopic changes in the kidney of rats exposed to 10, 100 or 1000 ppm allyl chloride which were considered treatment related.

- In the liver of rats from the 1000 ppm group there was a microscopic change that was readily detected and consisted of an apparent decrease in hepatic glycogen. As a result, the cytoplasm of the hepatocytes throughout the lobule appeared to stain more intensely eosinophilic and was without any evidence of a degenerative change. Similar findings were noted in the hepatocytes of the 100 ppm group but were of an equivocal degree. In the 10 ppm group of rats there were no microscopic differences in their liver when compared to their respective controls.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): bioaccumulation potential cannot be judged based on study results
F344 rats were treated via 3 different routes with 3-chloropropene. Blood levels as well as excretion parameters were determined. In addition the effect on the GSH level was determined.
3-chloropropene is readily absorbed via via the gasrointestinal tract as well as via inhalation, distributed quickly and extensively metabolised especially at lower doses (1 mg/Kg bw orally). At higher doses (100 mg/Kg bw) 3-chloropropene is exhaled to a significant amount unmetabolised. Primary routes of excreetion are the urine and the exhalation (either unchanged or as CO2). The half-life of 3 -chloropropene in blood after treatment with 100 mg/Kg via gavage is t1/2 = 2.58 h, after 100 mg/Kg via iv injection t1/2 = 23.5 min and after inhalation of 100 ppm for 6 h t1/2 = 16.7 min.
A 100 mg/kg oral dose would be approximately equivalent to to a six hour-150 ppm inhalation exposure and a 100 mg/kg intravenous dose would be approximately equivalent to a six hour-85 ppm exposure.
The inhalative exposure to 100 ppm or higher for 6 h lead to a significant decrease in non-protein sulfhydryl levels in several tissues that are indicative of an extensive reaction of 3-chloropropene and/or its metabolites with GSH.

Executive summary:

In the present study (Waechter 1982) F344 rats were treated via 3 different routes with 3-chloropropene. Blood levels as well as excretion parameters were determined. In addition the effect on the GSH level was determined.

3-chloropropene is readily absorbed via via the gasrointestinal tract as well as via inhalation (absorption rates for inhalation are 0.89, 9.83, 63.9 and 67.5 μg/min for 10, 100, 1000 and 2000 ppm for 6 h respectively), distributed quickly and extensively metabolised especially at lower doses (1 mg/Kg bw orally). After oral gavage with 1 mg/Kg bw at least 70 % absorption can be assumed while after treatmant with 100 mg/Kg bw at least 42.2 % absorption can be assumed (Tmax < 1 h and = 4 h respectively).

At higher doses (100 mg/Kg bw) 3-chloropropene is exhaled to a significant amount unmetabolised. Primary routes of excretion are the urine and the exhalation (either unchanged or as CO2). The half-life of 3 -chloropropene in blood after treatment with 100 mg/Kg via gavage is t1/2 = 2.58 h, after 100 mg/Kg via iv injection t1/2 = 23.5 min and after inhalation of 100 ppm for 6 h t1/2 = 16.7 min.

A 100 mg/kg oral dose would be approximately equivalent to to a six hour-150 ppm inhalation exposure and a 100 mg/kg intravenous dose would be approximately equivalent to a six hour-85 ppm exposure.

After oral application of 1 and 100 mg 3 -chloropropene, respectively 1.6 % and 0.9 % of the applied activity were still found after 48 hours in blood. Respectively 1.6 % and 3.7 % of the radioactivity were still found in the bled animal cadaver.

Based on this data a clear descision on bioaccumulation is not possible, but it can be expected to be neither neglibile nor extensive.

The inhalative exposure to 100 ppm or higher for 6 h lead to a significant decrease in non-protein sulfhydryl levels in several tissues that are indicative of an extensive reaction of 3-chloropropene and/or its metabolites with GSH.

The following methods were used.

Experiment A: single oral gavage of labelled test item at 1 and 100 mg/Kg, air trapping; urine and feces sampling (metabolic cages) at blood samples at 1, 2, 3, 4, 6, 8, 16, 24, 32, 40 and 48 h post dosing from indwelling jugular cannulas; determination of radioactivity in carcass and samples; sacrifice of animals 48 h post dosing and histopathologic analysis of liver and kidney tissue samples; bile cannulation of 2 rats; urinary metabolite profile determination from 2 animals

Experiment B: single oral gavage or single intravenous injection of non-labelled test item at 100 mg/Kg, blood sampling at 5, 10, 15, 30, 60, 120, and 180 minutes after dosing indwelling jugular cannulas.

Experiment C: Fasted rats exposed via inhalation at 10, 100, 1000 and 2000 ppm of unlabeled test item for 6 h in head-only exposure chambers; blood sampling at 0.25, 0.5, 1, 2, 4 and 6 h during exposure and at different time points thereafter; histopathologic analysis of liver and kidney tissue samples