The single oral dose LD50 for methylenediphenol in rats was found to be more than 2000 mg/kg for males and females.
Oral toxicity of methylenediphenol was studied in a 28-day repeat dose toxicity test in rats at dose levels of 0, 8, 40, 200 and 1000 mg/kg. One female of the 1000 mg/kg group was sacrificed upon becoming moribund. During the dosing period, decreased body weight and food consumption were detected in the 1000 mg/kg group. On blood chemical examination, decreased total cholesterol and glucose, as well as increased gGT in the 1000 mg/kg group, and decreased total cholesterol in the females of the 200 and 40 mg/kg groups, were observed at the end of the dosing period. An irritating effect of the test substance on the mucosa of digestive tract was suggested by the results of necropsy and histopathological examinations. Enzyme induction the by the test substance was suggested by the results of histopathological examination of the liver and measurement of liver weight. From this study, the NOELs are considered to be 40 mg/kg/day for males, and 8 mg/kg/day for females.
A reverse mutation test of methylenediphenol in bacteria was carried out. This substance was not mutagenic in Salmonella typhimurium TA100, TA1535, TA98, TA1537 and Escherichia coli WP2 uvrA, with or without an exogenous metabolic activation system.
Genotoxicity of methylenediphenol was studied by chromosomal aberration test using cultured Chinese hamster lung (CHL/IU) cells.
With short-term treatment, methylenediphenol induced structural chromosomal aberrations at the maximum doses of 0.0090 mg/mL and 0.090 mg/mL, with and without metabolic activation systems, respectively. No polyploidy was induced with any treatment.
| Purity | : | 99.0 % |
| Test species/strain | : | Rats/Crj:CD(SD)IGS |
| Test method | : | OECD Test Guideline 401 |
| Route | : | Oral(gavage) |
| Dosage | : | 0(vehicle), 2000 mg/kg |
| Number of animals/group | : | Males, 5; females, 5 |
| Vehicle | : | 0.5 % CMC-Na aqueous solution mixed with 0.1 % Tween 80 |
| GLP | : | Yes |
Test results:
In the other animals, no abnormalities were detected in terms of clinical signs, body weight, or necropsy.
The LD50 value was estimated to be more than 2000 mg/kg for males and females.
| Purity | : | 99.0 % |
| Test species/strain | : | Rat/Crj:CD(SD)IGS |
| Test method | : | Guideline for 28-Day Repeated Dose Toxicity Test in Mammalian Species (Chemical Substances Control Law of Japan) |
| Route | : | Oral(gavage) |
| Dosage | : | 0(vehicle), 8, 40, 200, 1000 mg/kg/day |
| Number of animals/group | : | Males, 12; females, 12(0, 200, 1000 mg/kg) Males, 6; females, 6(8, 40 mg/kg) |
| Vehicle | : | 0.5 % CMC-Na aqueous solution mixed with 0.1 % Tween 80 |
| Administration period | : | Males and females, 28 days |
| Terminal kill | : | Days 29 and 43 |
| GLP | : | Yes |
Test results:
Significant decrease in body weight in the males of the 1000 mg/kg group, and significant decrease in food consumption in both sexes of the 1000 mg/kg group were observed during the dosing period.
On blood chemical examination at the end of the dosing period, decreased total cholesterol in both sexes of the 1000 mg/kg group, decreased glucose and increased gGT in the females of the 1000 mg/kg group, and decreased total cholesterol in the females of the 200 and 40 mg/kg groups were observed. No changes were detected at the end of the recovery period.
On pathological examination, changes considered caused by the test substance were detected in the stomach and liver. Reddish change of glandular stomach wall, and thickening of the forestomach wall or limiting ridge in both sexes of the 1000 mg/kg group were found at necropsy. Histopathologically, squamous hyperplasia of the forestomach or limiting ridge, with ulceration of the forestomach were detected. An irritating effect of the test substance on the digestive tract was suggested.
On histopathological examination of liver, centrilobular hypertrophy of hepatocytes was found in both sexes of the 200 and 1000 mg/kg groups. Since increased liver weights in both sexes of the 1000 mg/kg group (absolute and/or relative liver weights), and in the females of the 200 mg/kg group (relative liver weights) were simultaneously observed, enzyme induction by the test substance was suggested.
No abnormalities were detected on hematological examination or urinalysis.
From this study the NOELs are considered to be 40 mg/kg/day for males, and 8 mg/kg/day for females.
| Purity | : | 99.0 % |
| Test species/strains | : | Salmonella typhimurium TA100, TA1535, TA98, TA1537, Escherichia coli WP2 uvrA |
| Test method | : | Guidelines for Screening Mutagenicity Testings of Chemicals(Chemical Substances Control Law of Japan) and OECD Test Guideline 471 |
| Procedures | : | Pre-incubation method |
| Solvent | : | Dimethyl sulfoxide |
| Positive controls | : | -S9 mix; 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (TA100, TA98, WP2 uvrA), Sodium azide (TA1535) and 9-Aminoacridine (TA1537) +S9 mix; 2-Aminoanthracene (five strains) |
| Doses | : | -S9 mix; 0, 39.1, 78.1, 156, 313, 625, 1250 μg/plate(five strains) +S9 mix; 0, 39.1, 78.1, 156, 313, 625, 1250 μg/plate(TA100, TA1535, TA1537) +S9 mix; 0, 78.1, 156, 313, 625, 1250, 2500 μg/plate(TA98) +S9 mix; 0, 156, 313, 625, 1250, 2500, 5000 μg/plate(WP2 uvrA) |
| S9 | : | Rat liver, induced with phenobarbital and 5,6-benzoflavone |
| Plates/test | : | 3(1 for cytotoxicity test) |
| Number of replicates | : | 2(plus 1 cytotoxicity test) |
| GLP | : | Yes |
Test results:
Genetic effects:
Salmonella typhimurium TA100, TA1535, TA98, TA1537
| + | ? | - | |
| Without metabolic activation: | [ ] | [ ] | [*] |
| With metabolic activation: | [ ] | [ ] | [*] |
Escherichia coli WP2 uvrA
| + | ? | - | |
| Without metabolic activation: | [ ] | [ ] | [*] |
| With metabolic activation: | [ ] | [ ] | [*] |
| Purity | : | 99.0 % |
| Type of cell used | : | Chinese hamster lung (CHL/IU) cells |
| Test method | : | Guidelines for Screening Mutagenicity Testing of Chemicals(Chemical Substances Control Law of Japan) and OECD Test Guideline 473 |
| Solvent | : | Dimethyl sulfoxide |
| Positive controls | : | -S9 mix; Mitomycin C +S9 mix; Cyclophosphamide |
| Doses | : | -S9 mix(short-term treatment); 0, 0.023, 0.045, 0.090 mg/mL +S9 mix(short-term treatment); 0, 0.0023, 0.0045, 0.0090 mg/mL -S9 mix(continuous treatment for 24 hr); 0, 0.015, 0.030, 0.060 mg/mL |
| S9 | : | Rat liver, induced with phenobarbital and 5,6-benzoflavone |
| Plates/test | : | 2 |
| GLP | : | Yes |
Test results:
| Lowest concentration producing cytogenetic effects in vitro: | ||
| Without metabolic activation (short-term treatment) | : | 0.090 mg/mL(clastogenicity) |
| With metabolic activation (short-term treatment) | : | 0.0090 mg/mL(clastogenicity) |
Genotoxic effects:
| clastogenicity | polyploidy | |||||
| + | ? | - | + | ? | - | |
| Without metabolic activation: | [*] | [ ] | [ ] | [ ] | [ ] | [*] |
| With metabolic activation: | [*] | [ ] | [ ] | [ ] | [ ] | [*] |
| 1) | The tests were performed by the Mitsubishi Chemical Safety Institute Ltd., Kashima Laboratory. 14 Sunayama, Hasaki-machi, Kashima-gun, Ibaraki, 314-0255, Japan. Tel +81-479-46-2871 Fax +81-479-46-2874 |
| 2) | The tests were performed by the Hatano Research Institute, Food and Drug Safety Center, 729-5 Ochiai, Hadano-shi, Kanagawa, 257-8523, Japan. Tel +81-463-82-4751 Fax +81-463-82-9627 |