A combined repeat dose and reproductive/developmental toxicity screening test was performed with oral administration of 4-methoxybenzaldehyde at doses of 0, 20, 100, 500 mg/kg. No animal died in any group. Temporary salivation, tendency for increase in body weight, increase in food consumption, decrease in platelet count, increase in the A/G ratio, GOT activity and inorganic phosphorus concentration, increase in liver weight and decrease in epididymidal weight were observed in males and females of the 100 and/or 500 mg/kg groups. In the 100 and/or 500 mg/kg groups, hyperplasia of squamous epithelium in the forestomach and centrilobular hypertrophys of hepatocyte were detected. The fertility index and the number of pups were decreased in the 500 mg/kg group. From the results, the no observed effect dose level (NOEL) for systemic toxicity of 4-methoxybenzaldehyde is considered to be 20 mg/kg/day in males and females, and NOELs for reproductive and developmental toxicity are considered to be 100 mg/kg/day in males and females, and 100 mg/kg/day in pups.
Reverse mutation assays using microorganisms (Salmonella typhimurium, Escherichia coli) were conducted to assess the potential of 4-methoxybenzaldehyde to induce gene mutations.
4-Methoxybenzaldehyde did not induce gene mutations in the bacteria under the conditions of this study.
In vitro chromosomal aberration tests using cultured cells (CHL/IU) were conducted to assess the potential of 4-methoxybenzaldehyde to induce chromosomal aberrations.
4-Methoxybenzaldehyde did not induce chromosomal aberrations in cultured cells under the conditions of this study.
| Purity | : | 99.9 % |
| Test species/strain | : | Rat/Crj:CD(SD)IGS |
| Test method | : | OECD Test Guideline 401 |
| Route | : | Oral(gavage) |
| Doses | : | Males, 0(vehicle), 1000, 2000 mg/kg Females, 2000 mg/kg |
| Number of animals/group | : | Males, 5; females, 5 |
| Vehicle | : | Corn oil |
| GLP | : | Yes |
Test results:
The LD50 value was concluded to be more than 2000 mg/kg for both sexes.
| Purity | : | 99.9 % |
| Test species/strain | : | Rat/Crj:CD(SD)IGS |
| Test method | : | OECD Guideline 422 |
| Route | : | Oral(gavage) |
| Doses | : | 0(control), 20, 100, 500 mg/kg/day |
| Number of animals/group | : | Males, 13; Females, 13 |
| Vehicle | : | Corn oil |
| Administration period | : | Males, 42 days Females, from 14 days before mating to day 4 of lactation |
| GLP | : | Yes |
Test results:
No animal died in any group. Temporary salivation after administration was observed in males and females of the 500 mg/kg group. Body weight tended to be increased in males and females of the 100 and 500 mg/kg groups. Increase in food consumption was observed in males of the 500 mg/kg and females of the 100 and 500 mg/kg groups. Decrease in platelets was observed in 500 mg/kg males and 100 and 500 mg/kg females. Hyperplasia of squamou count epithelium was detected in males and females given 100 or 500 mg/kg. In the 500 mg/kg group, the liver weight was increased in males and females. Histologically, centrilobular hypertrophy of hepatocytes was detected in these animals. On biochemical analysis, the A/G ratio, GOT activity and inorganic phosphorus concentration were found to be increased in males of the 500 mg/kg group.
II. Reproductive and developmental toxicity
In the 500 mg/kg group, the number of non-pregnant females was increased whereas all pairs copulated, and the fertility index was reduced. Epididymidal weight was decreased in males of 500 mg/kg group. The number of pups, the delivery index and the number of live pups were lower than in the controls. The compound showed no adverse effects in terms of estrous cycle, histopathological changes in reproductive organs, parturition and lactation, as well as duration of pregnancy, number of corpora lutea and implant rate at any dose level. The compound did not demonstrate any adverse effects on viability, sex ratio, body weight and morphological appearance of pups.
III. Evaluation
From the results the no observed effect dose level (NOEL) for toxicity of 4-methoxybenzaldehyde is considered to be 20 mg/kg/day in males and females, NOELs for reproductive and developmental toxicity are considered to be 100 mg/kg/day in males and females, and 100 mg/kg/day in pups. The target organs of 4-methoxybenzaldehyde are considered to be the stomach and liver.
| Purity | : | 99.9 % |
| Test species/strains | : | Salmonella typhimurium TA100, TA1535, TA98, TA1537, Escherichia coli WP2 uvrA |
| Test method | : | Guidelines for Screening Mutagenicity Testing of Chemicals(Chemical Substances Control Law of Japan) and OECD Test Guideline 471 |
| Procedures | : | Pre-incubation method |
| Solvent | : | DMSO |
| Positive controls | : | -S9 mix; 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (TA100, TA98 and WP2 uvrA), Sodium azide (TA1535) and
9-Aminoacridine hydrochloride (TA1537) +S9 mix; 2-Aminoanthracene (all strains) |
| Doses | : | -S9 mix; 39.1, 78.1, 156, 313, 625, 1250 μg/plate(TA100, TA1535, TA1537) -S9 mix; 78.1, 156, 313, 625, 1250, 2500, 5000 μg/plate(WP2 uvrA) -S9 mix; 156, 313, 625, 1250, 2500, 5000 μg/plate(TA98) +S9 mix; 156, 313, 625, 1250, 2500, 5000 μg/plate(TA100, TA1535, TA98, TA1537) +S9 mix; 39.1, 78.1, 156, 313, 625, 1250, 2500, 5000 μg/plate(WP2 uvrA) |
| S9 | : | Rat liver, induced with phenobarbital and 5,6-benzoflavone |
| Plates/test | : | 3 |
| Number of replicates | : | 2 |
| 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 | : | 99.9 % |
| 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 | : | DMSO |
| Positive controls | : | -S9 mix; Mitomycin C +S9 mix; Cyclophosphamide |
| Doses | : | -S9 mix(short-term treatment); 0, 341, 681, 1362 μg/mL +S9 mix(short-term treatment); 0, 341, 681, 1362 μg/mL -S9 mix(continuous treatment 24 hr); 0, 85.1, 170, 341, 681 μg/mL |
| S9 | : | Rat liver, induced with phenobarbital and 5,6-benzoflavone |
| Plates/test | : | 2 |
| GLP | : | Yes |
Test results:
Genetic effects:
| clastogenicity | polyploidy | |||||
| + | ? | - | + | ? | - | |
| Without metabolic activation: | [ ] | [ ] | [*] | [ ] | [ ] | [*] |
| With metabolic activation: | [ ] | [ ] | [*] | [ ] | [ ] | [*] |
| 1) | The tests were performed by the Hatano Research Institute, Food and Drug Safety Center, 729-5 Ochiai, Hadano-shi, Kanagawa, 257-0025, Japan. Tel +81-463-82-4751 Fax +81-463-82-9627 |
| 2) | The tests were performed by the Biosafety Research Center, Foods, Drugs and Pesticides(An-pyo Center), 582-2 Shioshinden, Fukude-cho, Iwata-gun, Shizuoka, 437-1213, Japan. Tel +81-538-58-1266 Fax +81-538-58-1393 |