2,4,6-Trinitrophenol

2,4,6-トリニトロフェノール


[CAS No. 88-89-1]

Picric acid

ピクリン酸

Molecular formula: C6H3N3O7　Molecular weight: 229.10

ABSTRACT

Single oral dose LD50 values of 2,4,6-trinitrophenol were found to be 492 mg/kg for males and 283 mg/kg for females.

2,4,6-Trinitrophenol was studied for oral toxicity in rats in a 28-day repeat dose toxicity test at 0, 4, 20 and 100 mg/kg.

Yellowish coloring of fur was observed in the 20 and 100 mg/kg groups, and yellowish chromaturia and transient salivation were noted in the 100 mg/kg group. Decreased food consumption was apparent for females of the 100 mg/kg group. Hematological examination revealed anemia in the 100 mg/kg group. Blood chemical examination revealed an increase in gGT and a decrease in glucose in the 100 mg/kg group. Urinalysis revealed a decrease in potassium in the 20 and 100 mg/kg groups. In the testis, diffuse atrophy of seminiferous tubules was observed in the 100 mg/kg group. In the cecum, ulceration was observed in the 100 mg/kg group. In the liver, hypertrophy of hepatocyte was observed in the 100 mg/kg group. The NOEL is considered to be 4 mg/kg/day for both sexes.

2,4,6-Trinitrophenol proved mutagenic in Salmonella typhimurium TA98 and TA1537 without an exogenous metabolic activation system, and in TA100, TA1535, TA98 and TA1537 with an exogenous metabolic activation system.

2,4,6-Trinitrophenol induced structural chromosomal aberrations in CHL/IU cells after short-term treatment without an exogenous metabolic activation system. Polyploidy was not induced in any treatment group.

SUMMARIZED DATA FROM THE STUDIES

1. Single Dose Toxicity 1)

Purity:81.4 %
Test species/strain:Rat/Crj:CD(SD)IGS
Test method:OECD Test Guideline 401
　Route:Oral(gavage)
　Dosage:Males, 0(vehicle), 200, 400, 800 mg/kg
females, 0(vehicle), 100, 200, 400, 800 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:

Deaths occurred in the 400 mg/kg and above dose groups for males and in the 200 mg/kg and more groups for females. Clinical signs observed were as follows: yellowish coloring of fur, yellowish chromaturia, decrease in locomotor activity, abnormal gait, yellowish coloring of skin, clonic convulsions, loose stool, soiled perineal region, adoption of a prone or lateral position, and reddish tears. The body weights of surviving animals increased in the same manner as those of the control group. Pathological changes in dead animals were observed in the glandular stomach along with yellowish coloring of the whole body. Yellowish tinction of fur was also evident in survivors.

The LD50 values were found to be 492 mg/kg for males and 283 mg/kg for females.

2. Repeat Dose Toxicity 1)

Purity:81.4 %
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), 4, 20, 100 mg/kg/day
　Number of animals/group:Males, 6; females, 6
　Vehicle:0.5 % CMC-Na aqueous solution mixed with 0.1 % Tween 80
　Administration period:Males and females, 28 days
　Sacrifice:Days 29 and 43
GLP:Yes

　Test results:

Yellowish coloring of fur was observed in both sexes of the 20 and 100 mg/kg groups, and yellowish chromaturia and transient salivation were observed in both sexes of the 100 mg/kg group. Food consumption was decreased in females of the 100 mg/kg group. Hematological examination revealed decreases in the erythrocyte count and Hb in both sexes, and MCHC in females and increases in reticulocyte and leukocyte counts in both sexes, and MCV in females of the 100 mg/kg group. Blood chemical examination revealed an increase in gGT in males and a decrease in glucose in females of the 100 mg/kg group. Urinalysis revealed a decrease in potassium in both sexes receiving 100 mg/kg and in females of the 20 mg/kg group. Organ weight measurement revealed increases in absolute and relative weights of the spleen and liver in both sexes, and decreases in absolute and relative weights of epididymides in the 100 mg/kg group. Necropsy revealed enlargement of the spleen, erosion/ulceration of the cecum, and yellowish chromaturia in urinary bladder in both sexes, and yellowish tinction of the testis in the 100 mg/kg group. Histopathological examination revealed altered development of germinal centers, hemosiderin deposition, and extramedullary hematopoiesis of the spleen, ulcers of the cecum, and centrilobular hypertrophy of hepatocytes in both sexes, and diffuse atrophy of seminiferous tubules of the testes, cell debris in the lumen and decrease in sperm of the epididymis in the 100 mg/kg group.

Yellowish coloring of fur, an increase in MCV and decreases in MCHC and absolute and relative weights of epididymides, and small testis, hemosiderin deposition in the spleen, focal hyperplasia of mucosal epithelium, granulation tissue of the cecum, diffuse atrophy of seminiferous tubules of the testis, cell debris in lumen and decrease in sperm of the epididymis were also observed after withdrawal.

The NOEL is considered to be 4 mg/kg/day for both sexes.

3. Genetic Toxicity

3-1. Bacterial test 1)

Purity:81.4 %
Test species/strain: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), Sodium azide (TA1535), 9-Aminoacridine (TA1537) and N-Ethyl-N'-nitro-N-nitrosoguanidine(WP2 uvrA)
+S9 mix; 2-Aminoanthracene(all strains)
　Doses:-S9 mix; 156, 313, 625, 1250, 2500 and 5000 μg/plate(TA100, TA1535 and TA98)
-S9 mix; 313, 625, 1250, 2500 and 5000 μg/plate(WP2 uvrA)
-S9 mix; 78.1, 156, 313, 625, 1250, 2500 and 5000 μg/plate(TA1537)
+S9 mix; 156, 313, 625, 1250, 2500 and 5000 μg/plate(TA100, TA1535 and TA98)
+S9 mix; 313, 625, 1250, 2500 and 5000 μg/plate(WP2 uvrA)
+S9 mix; 78.1, 156, 313, 625, 1250, 2500 and 5000 μg/plate(TA1537)
　S9:Rat liver, induced with phenobarbital and 5,6-benzoflavone
　Plates/test:3
　Number of replicates:2
GLP:Yes

　Test results:

This chemical induced gene mutations in Salmonella typhimurium TA98 and TA1537 without metabolic activation, and in TA100, TA1535, TA98 and TA1537 with metabolic activation. Toxicity was observed at 5000 μg/plate (TA100, TA1535 and TA98) without metabolic activation, and at 5000 μg/plate (TA1535) in the first main test and at 5000 μg/plate (TA100 and TA1535) in the second main test with metabolic activation.

Genetic effects:
Salmonella typhimurium TA100 and TA1535
+?-
　Without metabolic activation:[ ][ ][*]
　With metabolic activation:[*][ ][ ]

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

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

3-2. Non-bacterial in vitro test (chromosomal aberration test)1)

Purity:81.4 %
Type of cell used:Chinese hamster CHL/IU cells
Test method:Guidelines for Screening Mutagenicity Testing of Chemicals(Chemical Substances Control Law of Japan) and OECD Test Guideline 473
　Solvent:Acetone
　Positive controls:-S9 mix; Mitomycin C
+S9 mix; Benzo[a]pyrene
　Doses:-S9 mix(6 hr short-term treatment); 0, 200, 400, 800, 1600 μg/mL(main test)
-S9 mix(6 hr short-term treatment); 0, 1400, 1600, 1800 μg/mL(confirmation test)
+S9 mix(6 hr short-term treatment); 0, 200, 400, 800, 1600 μg/mL
　S9:Rat liver, induced with phenobarbital and 5,6-benzoflavone
　Plates/test:2
GLP:Yes

　Test results:

Structural chromosomal aberrations were induced at 1600 and 1800 μg/mL after 6 hr short-term treatment without S9 mix (19.0 and 21.0 %, respectively). Polyploidy was not induced in any treatment groups.

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

1)The tests were performed by the Kashima Laboratory, Mitsubishi Chemical Safety Institute Ltd., 14 Sunayama, Hasaki-machi, Kashima-gun, Ibaraki, 314-0255, Japan. Tel +81-479-46-2871 Fax +81-479-46-2874