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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
Repeated dose toxicity: oral
Administrative data
- Endpoint:
- chronic toxicity: oral
- Remarks:
- combined repeated dose and carcinogenicity
- 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
Data source
Reference
- Reference Type:
- publication
- Title:
- Safety evaluation of toothpaste containing chloroform III. Long-term study in Beagle dogs
- Author:
- Heywood R, Sortwell RJ, Noel PRB, Street AE, Prentice DE, Roe FJC, Wadsworth PF, Worden AN, Van Abbé NJ
- Year:
- 1 979
- Bibliographic source:
- Journal of Environmental Pathology and Toxicology 2: 835-851
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- EU Method B.33 (Combined Chronic Toxicity / Carcinogenicity Test)
- GLP compliance:
- no
- 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:
- dog
- Strain:
- Beagle
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- Pure bread dogs, initially 18 to 24 weeks old, were housed singly in kennels. All the dogs were clinically examined and inoculated against distemper, canine hepatitis and leptospirosis as well as being dose with piperazine adipate as an anthelmintic. Inoculatin and anthelmintic treatment were repeated annually. The dogs were fed weighed amounts (200 g) of a dry diet at fixed times twice daily. When new food was offered, any residue from the previous meal was removed. From week 300, the daily food ration of dogs considered to be obese was reduced from 400 to 300 g. Water was freely available at all times and fresh milk (200 mL) offered to each dog on weekday mornings for the first 6 months.
Administration / exposure
- Route of administration:
- oral: capsule
- Vehicle:
- other: toothpaste base (including peppermint oil and eucalyptol) given orally by gelatin capsule
- Details on oral exposure:
- Dogs received a dose of chloroform contained in tooth paste for 7 days per week during the main study.
- Analytical verification of doses or concentrations:
- no
- Duration of treatment / exposure:
- 7.5 years
- Frequency of treatment:
- 6 days per week
Doses / concentrations
- Remarks:
- Doses / Concentrations:0, 15 and 30 mg/kg body weigth/dayBasis:other: nominal in capsule
- No. of animals per sex per dose:
- 8 males, 8 females in the 30 and 15 mg/kg body weight dose groups and in untreated and alternative non-chloroform toothpaste groups; 16 males, 16 females in the 0 mg/kg body weight dose group
- Control animals:
- yes, concurrent no treatment
- yes, concurrent vehicle
- Details on study design:
- From observations made in preliminary studies and to ensure that the upper dose level would reach the threshold for early toxic effects but with many dogs as possible remaining alive and well for at least 7 years
Examinations
- Observations and examinations performed and frequency:
- Clinical signs: daily; food consumption: daily; water consumption: intermittently; body weight: once per week; opthalmoscopy: 3-month intervals; thorough clinical examination: at least twice yearly; comprehensive range of laboratory investigations: during pre-treatment period, after 6, 13 weeks, 6.9 and 12 months, then every 6 months; serum enzyme studies: in the later stages of the treatment;
- Sacrifice and pathology:
- Between weeks 395 and 399, each dog still alive received an intravenous injection of sodium pentobarbitone; immediately after killing or as soon as possible in the case of animals found dead, full macroscopic examination was made of all tissues and any abnormalities were noted; principal organs were removed and weighed; small portions of these and a wide range of other tissues along with portions of any lump or tumour were placed in fixative; sections were stained with haemotoxylin and eosin; examination, by electron microscopy, was conducted on liver and kidney sections
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):
- no effects observed
- Food efficiency:
- not specified
- Water consumption and compound intake (if drinking water study):
- not specified
- Ophthalmological findings:
- no effects observed
- Haematological findings:
- effects observed, treatment-related
- Clinical biochemistry findings:
- no effects observed
- Urinalysis findings:
- no effects observed
- Behaviour (functional findings):
- no effects observed
- Organ weight findings including organ / body weight ratios:
- no effects observed
- Gross pathological findings:
- no effects observed
- Histopathological findings: non-neoplastic:
- effects observed, treatment-related
- Histopathological findings: neoplastic:
- no effects observed
Effect levels
- Dose descriptor:
- LOAEL
- Effect level:
- 15 mg/kg bw/day (nominal)
- Sex:
- male/female
- Basis for effect level:
- other: increased serum levels of alanine aminotransferase and increased incidence of aggregations of vacuolated histiocytes forming "fatty cysts" in the liver of treated dogs
Target system / organ toxicity
- Critical effects observed:
- not specified
Any other information on results incl. tables
Table 1: Group mean values in biochemical analyses of different groups treated with chloroform, untreated control groups and groups receiving alternative toothpaste (male and female dogs combined)
Treatment (mg/kg/d) |
30 |
15 |
Vehicle control |
Untreated |
Alternative toothpaste |
|||||
No. of dogs |
15 |
15 |
27 |
12 |
15 |
|||||
Week |
374 a) |
395 b) |
374 |
395 |
374 |
395 |
374 |
395 |
374 |
395 |
Urea (mg %) |
30 |
28 |
32 |
26 |
31 |
31 |
33 |
27 |
34 |
29 |
Glucose (mg %) |
99 |
99 |
100 |
100 |
101 |
98 |
99 |
95 |
102 |
98 |
Total serum proteins (g %) |
5.9 |
6.1 |
6.1 |
6.2 |
6.3 |
6.0 |
6.3 |
6.0 |
6.4 |
6.1 |
Albumin |
2.9 |
2.8 |
3.0 |
2.8 |
3.1 |
2.8 |
3.2 |
2.9 |
3.2 |
2.9 |
alpha1 globulin |
0.3 |
0.3 |
0.3 |
0.3 |
0.3 |
0.4 |
0.3 |
0.3 |
0.3 |
0.3 |
alpha2 globulin |
0.7 |
0.8 |
0.7 |
0.8 |
0.7 |
0.8 |
0.7 |
0.7 |
0.7 |
0.8 |
beta globulin |
1.6 |
1.7 |
1.7 |
1.7 |
1.7 |
1.6 |
1.6 |
1.6 |
1.7 |
1.6 |
gamma globulin |
0.5 |
0.5 |
0.5 |
0.5 |
0.5 |
0.5 |
0.5 |
0.4 |
0.6 |
0.4 |
A/G ratio |
0.98 |
0.86 |
0.98 |
0.87 |
0.99 |
0.91 |
1.04 |
0.97 |
1.00 |
0.94 |
SAP (KA units) |
30 |
26 |
20 |
17 |
14 |
16 |
12 |
14 |
10 |
13 |
SGPT mU/mL |
102 |
111 |
66 |
48 |
51 |
128 |
50 |
56 |
57 |
87 |
SGOT mU/mL |
33 |
33 |
27 |
29 |
25 |
32 |
22 |
32 |
25 |
31 |
LAP (GR units) |
90 |
72 |
82 |
62 |
73 |
54 |
81 |
56 |
82 |
60 |
Bilirubin (mg %) |
0.2 |
0.2 |
0.2 |
0.2 |
0.2 |
0.2 |
0.2 |
0.2 |
0.2 |
0.2 |
Erythrocyte cholinesterase (delta pH/h) |
0.73 |
0.73 |
0.78 |
0.81 |
0.77 |
0.81 |
0.77 |
0.82 |
0.65 |
0.71 |
Plasma (delat pH/h) |
1.08 |
0.91 |
1.00 |
0.88 |
1.00 |
0.87 |
1.02 |
0.86 |
1.05 |
0.91 |
gamma GT (mU/mL) |
3.6 |
3.3 |
2.9 |
2.5 |
2.9 |
2.1 |
2.5 |
1.7 |
2.8 |
1.8 |
GDH (mU/mL) |
7.2 |
5.3 |
6.5 |
3.9 |
4.9 |
4.0 |
3.9 |
3.0 |
4.9 |
3.0 |
ICD (B&B units) |
11.8 |
11.4 |
11.2 |
10.7 |
9.9 |
12.9 |
9.2 |
11.7 |
10.6 |
12.9 |
a) end of treatment; b) end of recovery period; A/G: Albumin/Globulin; SAP: Serum Amyloid P component; SGPT: Serum Glutamic Pyruvic Transaminase (also Alanine Aminotransferase); SGOT: Serum Glutamic Oxaloacetic Transaminase (also Aspartate Transaminase); gamma GT (Gamma Glutamyl Transferase); GDH: Glutamate Dehydrogenase; ICD: Isocitrate Dehydrogenase
Table 2: Changes of alanine aminotransferase (ALAT) levels throughout the main study
Treatment (mg/kg/d) |
Group mean ALAT (mU/mL) |
||||||||||||||||||||
Pre-reatment |
Posttreatment (weeks) |
||||||||||||||||||||
6 |
13 |
26 |
39 |
52 |
78 |
104 |
130 |
156 |
182 |
208 |
234 |
260 |
286 |
312 |
338 |
364 |
372 |
14 |
19 |
||
30 mg/kg/d |
24 |
34 a) |
37 b) |
58 c) |
63 c) |
52 c) |
73 c) |
64 c) |
51 c) |
76 c) |
108 c) |
91 c) |
80 c) |
147 c) |
138 c) |
128 c) |
134 c) |
104 c) |
102 c) |
105 c) |
111 |
15 mg/kg/d |
22 |
29 |
30 |
33 |
39 |
32 |
43 |
45 |
34 b) |
46 b) |
61 b) |
55 b) |
55 a) |
95 c) |
93 b) |
89 c) |
73 a) |
79 a) |
66 |
53 |
48 |
Vehicle control |
22 |
29 |
29 |
30 |
39 |
29 |
35 |
40 |
22 |
30 |
33 |
40 |
34 |
33 |
65 |
47 |
49 |
50 |
51 |
56 |
128 |
Alternative toothpaste |
28 |
31 |
31 |
33 |
40 |
28 |
37 |
36 |
21 |
30 |
34 |
36 |
35 |
50 |
47 |
52 |
47 |
59 |
57 |
48 |
87 |
Untreated |
24 |
30 |
30 |
30 |
38 |
27 |
37 |
37 |
21 |
29 |
33 |
30 |
33 |
32 |
48 |
50 |
44 |
50 |
50 |
53 |
56 |
a) comparison with untreated group; p 0.05; b) comparison with untreated group; p 0.01; c) comparison with untreated group; p 0.001
Table 3: Liver changes (nodules of altered hepatocytes and “fatty cysts”)
Treatment (mg/kg/d) |
Sex |
No. of dogs examined histologically at end of experiment |
No. with nodules |
No. with “fatty cysts” |
|
Occasional or minimal |
Moderate or marked |
||||
30 |
Male |
7 |
0 |
1 |
6 |
Female |
8 |
4 |
0 |
7 |
|
15 |
Male |
7 |
1 |
0 |
6 |
Female |
8 |
1 |
2 |
3 |
|
Vehicle control |
Male |
15 |
0 |
7 |
1 |
Female |
12 |
3 |
3 |
0 |
|
Untreated |
Male |
7 |
1 |
2 |
0 |
Female |
5 |
1 |
1 |
0 |
|
Alternative toothpaste |
Male |
8 |
0 |
2 |
0 |
Female |
7 |
1 |
0 |
0 |
Applicant's summary and conclusion
- Conclusions:
- This long-term study investigating chronic effects of chloroform applied orally to dogs provided a LOAEL value of 15 mg/kg body weight/day.
- Executive summary:
A combined repeated dose and carcinogenicity study was carried out with chloroform orally administered to male and female Beagle dogs over a period of more than 7 years. The study was comparable to the principles set out in the testing guideline for method B.33 suggested by the European Commission with minor restrictions. The chloroform was contained in toothpaste and administered in form of a gelatine capsule.
Chronic effects related to chloroform exposure included a significantly increased level of alanine aminotransferase level in the high dose group throughout the study and in the lower-dose group during the later phase of the study and increased incidence of aggregations of vacuolated histiocytes forming "fatty cysts" in the liver of treated dogs.
A NOAEL value could not be established and the LOAEL value found in this study was 15 mg/kg body weight/day established for female and male dogs together.
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