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EC number: 212-742-4 | CAS number: 865-49-6
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Toxicity to reproduction
Administrative data
- Endpoint:
- two-generation reproductive toxicity
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
Cross-reference
- Reason / purpose for cross-reference:
- reference to same study
Data source
Reference
- Reference Type:
- publication
- Title:
- Unnamed
- Year:
- 1 997
Materials and methods
- Principles of method if other than guideline:
- Reproductive Assessment by Continuous Breeding (RACB) protocol of the US National Toxicology Program (NTP) according to Lamb (1985), J. Amer. Coll. Toxicol. 4, 163-171 and Reel et al. (1985), J. Amer. Coll. Toxicol. 4, 147-162
- GLP compliance:
- yes
- Limit test:
- no
Test material
- Reference substance name:
- Chloroform
- EC Number:
- 200-663-8
- EC Name:
- Chloroform
- Cas Number:
- 67-66-3
- Molecular formula:
- CHCl3
- IUPAC Name:
- chloroform
- Test material form:
- liquid
Constituent 1
Test animals
- Species:
- mouse
- Strain:
- other: CD-1 (ICR)BR
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS- Source: Charles River Breeding Laboratories Inc., Kingston, New York- Age at study initiation: 11 weeks- Weight at study initiation: (P) Males: 31.96-38.92 g; Females: 23.18-29.66 g; (F1) Males: 32.68 +/- 0.52 g; Females: 26.56 +/- 0.55 g- Housing: 2 per cage by sex during quarantine and the 1-week pre-mating using solid bottom polycarbonate cages with stainless steel wire lids; animals were subsequently housed as breeding pairs or individually- Diet (e.g. ad libitum): NIH-07 diet ad libitum- Water (e.g. ad libitum): ad libitum- Acclimation period: 2 weeks of quarantine, 1 week of acclimationENVIRONMENTAL CONDITIONS- Temperature (°C): 18 to 26- Humidity (%): no data- Air changes (per hr): no data- Photoperiod (hrs dark / hrs light): 14 hours light/10 hours darkness
Administration / exposure
- Route of administration:
- oral: gavage
- Vehicle:
- corn oil
- Details on exposure:
- Animals were treated with 8, 20 or 50 mg/kg body weight/day by gavage. The test chemical was mixed with Mazola corn oil on a weight to volume basis. Corn oil was tested for peroxide content prior to formulation and discarded if peroxide level was greater 10 meq. Each dose level was independently formulated. The concentration of chloroform was adjusted so that all doses were administered in 10 mL/kg body weight. Dose formulations were prepared at minimum every three weeks and aliquotted at the time of formulation into vials for daily use. Aliquots were stored at approximately 4 °C until use.
- Details on mating procedure:
- continuous cohabitation phase (14 weeks)
- Analytical verification of doses or concentrations:
- yes
- Details on analytical verification of doses or concentrations:
- An aliquot of each formulation, the control and the bulk chemical were sent to Research Triangle Institute for reference analysis during weeks 1, 6, 10 and 14 of the F1 study. Dosage formulation studies performed by Research Triangle Institute indicated no problems with the preparation of corn oil solutions at the 40 mg/mL level. Stability studies on corn oil solutions of chloroform (5 mg/mL level) indicated no significant loss of chemical after three weeks storage in sealed glass bottles in the dark at room temperature. Analysis of dosing solutions by gas chromatography showed that the actual doses administered to the animals were closer to 6.6, 15.9 and 41.2 mg/kg body weight/day.
- Duration of treatment / exposure:
- Continuous for 18 weeks (1 week prior to cohabitation, 14 weeks of cohabitation 3 weeks therefater) : Treatment of F0 and F1 generations
- Frequency of treatment:
- daily
- Details on study schedule:
- At the end of the dosing phase, the last litter from all dose groups was reared by the dam until weaning. At weaning, the F0 animals were killed. The pubs from the low and middle dose groups were killed and the pubs from the control and high dose groups were reared and dosed through the mating period (at approximately postnatal day 74) until necropsy.
Doses / concentrations
- Remarks:
- Doses / Concentrations:0, 6.6, 15.9 and 41.2 mg/kg body weight/dayBasis:analytical conc.
- No. of animals per sex per dose:
- 20 breeding pairs for dose groups, 40 breeding pairs for control
- Control animals:
- yes, concurrent vehicle
- Details on study design:
- -Dose selection rationale: Data on body weights, clinical signs, and food and water consumption from a 2-week dose-range-finding study.
Examinations
- Parental animals: Observations and examinations:
- Clinical signs, parental body weight and average consumption of drinking water during representative weeks
- Oestrous cyclicity (parental animals):
- As monitored by vaginal lavage for the 12 days preceding necropsy
- Sperm parameters (parental animals):
- Epididymal sperm motility, sperm morphology, sperm count
- Litter observations:
- Litters per pair, live pups per litter, proportion of pups born alive, sex of live pups and the pup body weights
- Postmortem examinations (parental animals):
- Selected organs: testes, epididymides and ovaries
- Statistics:
- The Cochran-Armitage test (Armitage 1971, Statistical Methods in Medical Research, John Wiley & Sons, New York) was used to tes for a dose-related trend in fertility. Dose group means for the number of litters, the number of live pups per litter, the proportion of live pups (number of pups born alive divided by the total number of pups produced by each pair), and the sex ratio (total number of male pups born alive out of the total number of live pups born to each fertile pair) were tested for overall differences using the Kruskal-Wallis test and for ordered differences using Jonckheere's test. Pairwise comparisons of treatment group means were performed by applying the Wilcox-Mann-Whitney U test. To remove the potential effect of the number of pups per litter on the average pup weight, an analysis of covariance (Neter and Wasserman 1974, Applied Linear Statistical Models) was performed. Least squares estimates of dose group means, adjusted for litter size, were computed and tested for overall equality using an F-test and pairwise equality using a t-test (carried out for males, females and both sexes combined). An analysis of covariance was used to adjust organ weights for total body weight. Unadjusted body and organ weights were analysed using the Kruskal-Wallis and Wilcox-Mann-Whitney U tests. Dose-related trends were tested for by Jonckheere's test.
Results and discussion
Results: P0 (first parental generation)
General toxicity (P0)
- Clinical signs:
- no effects observed
- Body weight and weight changes:
- no effects observed
- Food consumption and compound intake (if feeding study):
- no effects observed
- Organ weight findings including organ / body weight ratios:
- not examined
- Histopathological findings: non-neoplastic:
- not examined
- Other effects:
- no effects observed
Reproductive function / performance (P0)
- Reproductive function: oestrous cycle:
- no effects observed
- Reproductive function: sperm measures:
- no effects observed
- Reproductive performance:
- no effects observed
Details on results (P0)
Effect levels (P0)
- Key result
- Dose descriptor:
- NOAEL
- Effect level:
- 41.2 mg/kg bw/day (nominal)
- Based on:
- test mat.
- Sex:
- male/female
- Remarks on result:
- not determinable due to absence of adverse toxic effects
Results: F1 generation
General toxicity (F1)
- Clinical signs:
- no effects observed
- Mortality / viability:
- no mortality observed
- Body weight and weight changes:
- no effects observed
- Sexual maturation:
- no effects observed
- Organ weight findings including organ / body weight ratios:
- effects observed, treatment-related
- Gross pathological findings:
- effects observed, treatment-related
- Histopathological findings:
- effects observed, treatment-related
Details on results (F1)
Effect levels (F1)
- Key result
- Dose descriptor:
- NOAEL
- Generation:
- F1
- Effect level:
- 15.9 mg/kg bw/day (actual dose received)
- Sex:
- male/female
- Basis for effect level:
- other: Based on increased fertility index, relative epididymis weight and liver weight
Overall reproductive toxicity
- Key result
- Reproductive effects observed:
- no
Any other information on results incl. tables
Table 1: Reproductive toxicity of chloroform in Swiss mice
Reproductive toxicity in F0 generation |
7 mg/kg Male, female |
16 mg/kg Male, female |
41 mg/kg Male, female |
Average # litters/pair |
No change |
No change |
No change |
# live pubs/litter; pub weight/litter |
No change |
No change |
No change |
Cumulative days to litter |
No change |
No change |
No change |
Absolute testis, epididymis weight a) |
No observation |
No observation |
No observation |
Sex accessory gland weight a) (prostate, seminal vesicle) |
No observation |
No observation |
No observation |
Epidid. Sperm parameters (#, motility, morphology) |
No observation |
No observation |
No observation |
Estrous cycle length |
No observation |
No observation |
No observation |
Reproductive toxicity in F1 generation |
7 mg/kg Male, female |
16 mg/kg Male, female |
41 mg/kg Male, female |
Fertility index |
No observation |
No observation |
Increase b) |
# live pubs/litter; pub weight/litter |
No observation |
No observation |
Increase b), No change |
Absolute testis, epididymis weight a) |
No observation |
No observation |
No change, Increase (b) |
Sex accessory gland weight a) (prostate, seminal vesicle) |
No observation |
No observation |
No change |
Epidid. Sperm parameters (#, motility, morphology) |
No observation |
No observation |
No change |
Estrous cycle length |
No observation |
No observation |
No observation |
a) adjusted to bodyweight; b) statistically significant change (p 0.05)
Table 2: General toxicity of chloroform in Swiss mice
General toxicity in the F0 generation |
7 mg/kg Male, female |
16 mg/kg Male, female |
41 mg/kg Male, female |
Body weight |
No change |
No change |
No change |
Kidney weigh a) |
No observation |
No observation |
No observation |
Liver weight a) |
No observation |
No observation |
No observation |
Mortality |
No change |
No change |
No change |
Feed consumption |
No observation |
No observation |
No observation |
Water consumption |
No change |
No change |
No change |
Clinical signs |
No change |
No change |
No change |
General toxicity in the F1 generation |
7 mg/kg Male, female |
16 mg/kg Male, female |
41 mg/kg Male, female |
Pub growth to weaning |
No observation |
No observation |
No change |
Mortality |
No observation |
No observation |
No change |
Adult body weight |
No observation |
No observation |
No change |
Kidney weight a) |
No observation |
No observation |
No change |
Liver weight a) |
No observation |
No observation |
No change, Increase b) |
Feed consumption |
No observation |
No observation |
No observation |
Water consumption |
No observation |
No observation |
No change |
Clinical signs |
No observation |
No observation |
No change |
a) adjusted to bodyweight; b) statistically significant change (p 0.05)
Applicant's summary and conclusion
- Executive summary:
A two-generation reproductive toxicity test was carried out with chloroform using Swiss mice according to the Reproductive Assessment by Continuous Breeding (RACB) protocol. Chloroform was administered by oral gavage at actual doses of 0, 6.6, 15.9 and 41.2 mg/kg bodyweight/day.
The second generation reproductive assessments were conducted using pups from the control and high dose (41.2 mg) groups only. Of the 20 cohabited control pairs, 14 delivered live pups, while 19 of 20 high dose pairs delivered live pups. The controls delivered 10 pups per litter, the treated groups 12 pups. There were no other differences between the groups.
The necropsy of the F1 parent generation showed that there was no difference in female body weights, while female body weight-adjusted liver weight was elevated by 14 % in the treated group. All females showed some degree of hepatocellular degeneration. In males, the only difference between the control and the high dose groups was a 7 % increase in relative epididymis weight of the treated animals. There were no differences in epididymal sperm measures.
In summary, based on the findings, the NOAEL of the study for the reproduction functions was 15.9 mg/kg body weight/day.
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