4-Nitro-o-anisidine

4-ニトロ-o-アニシジン


[CAS No. 97-52-9]

2-Amino-5-nitroanisole/2-Methoxy-4-nitroaniline

2-アミノ-5-ニトロアソール/2-メトキシ-4-ニトロアニリン

Molecular formula: C7H8N2O3　Molecular weight: 168.17

ABSTRACT

4-Nitro-o-anisidine was studied for oral toxicity in rats in a 28-day repeat dose toxicity test at doses of 0, 30, 100 and 300 mg/kg. Decreases in hematocrit levels, hemoglobin levels, red blood cell counts, and increases in platelet and reticulocyte counts were observed in both males and females receiving 300 mg/kg. Increases in total cholesterol, GOT, GPT and total birilubin levels were also noted in both sexes receiving 300 mg/kg, along with elevated absolute and relative spleen, and relative liver weights, and slightly decreased absolute thymus weight. Histopathologically, increases in myocardial necrosis and extramedullary hematopoiesis in the spleen were observed in both males and females receiving 300 mg/kg. The NOEL for repeat dose toxicity is considered to be 30 mg/kg/day for both sexes.

4-Nitro-o-anisidine was mutagenic in Salmonella typhimurium TA100, TA98 and TA1537, with and without an exogeneous metabolic activation system and Salmonella typhimurium TA100, TA1535, TA98 and TA1537 and Escherichia coli WP2 uvrA, with an exogeneous metabolic activation system.

Genotoxicity of 4-nitro-o-anisidine was studied by chromosomal aberration test in cultured Chinese hamster lung (CHL/IU) cells. Structural chromosomal aberrations were induced at 0.080 mg/ml (high concentration) with short-term treatment and an exogenous metabolic activation system. Polyploidy was not induced in any treatment group.

SUMMARIZED DATA FROM THE STUDIES

1. Repeat Dose Toxicity１）

Purity:99.4%
Test species/strain:Rat/Crj:CD (SD)
Test method:Guidelines for 28-day Repeat Dose Toxicity Testing of Chemicals (Japan)
　Route:Oral (gavage)
　Doses:0 (vehicle), 30, 100, 300 mg/kg/day
　Number of animals:Male, 5; Female, 5/group
　Vehicle:Corn oil
　Administration period:Males and females, 28 days
　Terminal kill:Males and females, Days 29 or 43
GLP:Yes
　Test results:

Decreased spontaneous motor activity was observed in both males and females receiving 100 and 300 mg/kg, along with, subnormal temperature and salivation at the higher dose. Mean body weights of both males and females receiving 300 mg/kg were slightly less than those of the controls. Hematocrit and hemoglobin levels and red blood cell, platelet and reticulocyte counts were increased in both males and females receiving 300 mg/kg. Moreover, hematocrit levels of 100 mg/kg females were less than those of the controls. On blood chemistry examination, total cholesterol, GOT, GPT and total birilubin levels in both males and females receiving 300 mg/kg were greater than those of the controls.

Absolute and relative spleen and relative liver weights were increased in both males and females receiving 300 mg/kg, and absolute thymus weights were slightly decreased. Histopathological examination revealed increases in myocardial necrosis and extramedullary hematopoiesis in the spleen in both males and females receiving 300 mg/kg. Deposits of pigment in the spleen in females receiving 300 mg/kg were greater than in the controls.

The NOEL for repeat dose toxicity is considered to be 30 mg/kg/day for both sexes.

2. Genetic Toxicity

2-1. Bacterial test２）

Purity:99.4 wt%
Test species/strains:Salmonella typhimurium, TA100, TA1535, TA98, TA1537, Escherichia coli WP2 uvrA
Test method:Guidelines for Screening Mutagenicity Testing of Chemicals (Japan) and OECD Guideline No. 471 and 472
　Procedures:Pre-incubation method
　Solvent:DMSO
　Positive controls:-S9 mix, 2-(2-Furyl)-3-(5-nitro-2-furyl) acrylamide (TA100, TA98, WP2), Sodium azide (TA1535) and 9-Aminoacridine (TA1537)
+S9 mix, 2-Aminoanthracene (five strains)
　Doses:-S9 mix; 0, 313, 625, 1250, 2500, 5000 μg/plate (five strains)
+S9 mix; 0, 313 - 5000 μg/plate (five strains) and
　S9:Rat liver, induced with phenobarbital and 5,6-benzoflavone
　Plates/test:3
　Number of replicates:2
GLP:Yes

　Test results:

This chemical induced mutations in the S. typhimurium TA100, TA1535, TA98, TA1537 and E. coli strains with an S9 mix, and in TA100, TA98 and TA1537 without an S9 mix. Toxicity was not observed at 5000 μg/plate in the five strains with or without an S9 mix.

Genotoxic effects:

Salmonella typhimurium TA100, TA98, TA1537
+?-
Without metabolic activation:[*][ ][ ]
With metabolic activation:[*][ ][ ]

Salmonella typhimurium TA1535
+?-
Without metabolic activation:[ ][ ][*]
With metabolic activation:[*][ ][ ]

Escherichia coli WP2 uvrA
+?-
Without metabolic activation:[ ][ ][*]
With metabolic activation:[*][ ][ ]

2-2. Non-bacterial in vitro test (chromosomal aberration test)２）

Purity:99.4 wt%
Type of cell used:Chinese hamster lung (CHL/IU) cells
Test method:Guidelines for Screening Mutagenicity Testing of Chemicals (Japan) and OECD Guideline No. 473
　Solvent:Dimethylsulfoxide
　Positive controls:-S9 mix, Mitomycin C
+S9 mix, Cyclophosphamide
　Doses:-S9 mix (continuous treatment):0, 0.050, 0.10, 0.20 mg/m
-S9 mix (short-term treatment):0, 0.43, 0.85, 1.70 mg/ml
+S9 mix (short-term treatment):0, 0.020, 0.040, 0.080 mg/ml
　S-9:Rat liver, induced with phenobarbital and 5,6-benzoflavone
　Plates/test:2
GLP:Yes

　Test results:

Cytogenetic effects were seen as follows.

With short-term treatment and exogenous metabolic activation, structural chromosomal aberrations (12.0%, including gap) were induced at 0.080 mg/ml (high concentration). Polyploidy was not induced in any treatment groups.

Lowest concentration producing cytogenetic effects in vitro:

With metabolic activation (short-term treatment): 0.080 mg/ml (clastogenicity)

　Genotoxic effects:

clastogenicitypolyploidy
+?- +?-
Without metabolic activation:[ ][ ][*][ ][ ][*]
With metabolic activation:[*][ ][ ][ ][ ][*]

1)The tests were performed by the Biosafety Research Center, Foods, Drugs and Pesticides (An-pyo Center), Japan, 582-2 Shioshinden Arahama, Fukude-cho, Iwata-gun, Shizuoka, 437-12, Japan. Tel +81-538-58-1266 Fax +81-538-58-1393
2)The tests were performed by the Hatano Research Institute, Food and Drug Safety Center, 729-5 Ochiai, Hadano-shi, Kanagawa, 257, Japan. Tel +81-463-82-4751 Fax +81-463-82-9627