Pigment orange 16

ピグメントオレンジ16


[CAS No. 6505-28-8]

Dianisidine orange

ジアニシジンオレンジ

Molecular formula: C34H32N6O6　Molecular weight: 620.65

ABSTRACT

A single dose oral toxicity test of pigment orange 16 revealed an LD50 value of more than 2000 mg/kg for both sexes.

Pigment orange 16 was studied for oral toxicity in rats in a 28-day repeat dose toxicity test at doses of 0, 100, 300 and 1000 mg/kg. No deaths were observed in either sex. Repeated administration at 1000 mg/kg did not affect physiological function or morphological features. Thus, the NOEL for the 28-day repeat dose oral toxicity test of pigment orange 16 is considered to be 1000 mg/kg/day for males and females.

Reverse mutation assays using microorganisms (Salmonella typhimurium, Escherichia coli) were conducted to assess the potential of pigment orange 16 to induce gene mutations.

Pigment orange 16 did not induce gene mutations in bacteria under the conditions of this study.

In vitro chromosomal aberration tests using cultured cells (CHL/IU) were conducted to assess the potential of pigment orange 16 to induce chromosomal aberrations.

Pigment orange 16 did not induce chromosomal aberrations in cultured cells under the conditions of this study.

SUMMARIZED DATA FROM THE STUDIES

1. Single Dose Oral Toxicity 1)

Purity:>99 %
Test species/strain:Rat/Crj:CD(SD)IGS
Test method:OECD Test Guideline 401
　Route:Oral(gavage)
　Doses:Males and females, 0(vehicle), 2000 mg/kg
　Number of animals/group:Males, 5; females, 5
　Vehicle:Corn oil
GLP:Yes

　Test results:

No deaths occurred in any group. Mucous feces attributable to the administration of corn oil were observed in controls on the administration day and orange colored feces in both sexes of the treated group on the next day. No effects were found on body weights or autopsy findings.

The LD50 value was concluded to be more than 2000 mg/kg for both sexes.

2. Repeat Dose Toxicity 1)

Purity:>99 %
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)
　Doses:0(vehicle), 100, 300, 1000 mg/kg/day
　Number of animals/group:Males, 10; females, 10(0, 1000 mg/kg)
Males, 5; females, 5(100, 300 mg/kg)
　Vehicle:Corn oil
　Administration period:Males and females, 28 days
　Terminal kill:Males and females, on days 29 and 43
GLP:Yes

　Test results:

In the repeat dose study, no deaths were observed in either sex. The animals in the test substance groups excreted orange-colored feces due to the test substance continuously from the 2nd day to the end of administration period, but no other changes attributable to the test substance administered were observed regarding the general condition of the animals. There were no particular changes thought to have relevance to the test substance administration in parameters for body weight, food consumption, urinalysis, hematology, blood chemistry, organ weight and pathology.

Thus, the NOEL for the 28-day repeat dose toxicity is considered to be 1000 mg/kg/day for males and females.

3. Genetic Toxicity

3-1. Bacterial test 2)

Purity:99 % or more
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; 0, 4.88, 9.77, 19.5, 39.1, 78.1, 156, 313 μg/plate(TA100, TA1535, TA98, TA1537)
-S9 mix; 0, 39.1, 78.1, 156, 313, 625, 1250, 2500, 5000 μg/plate(WP2 uvrA)
+S9 mix; 0, 4.88, 9.77, 19.5, 39.1, 78.1, 156, 313 μg/plate(TA100, TA1535, TA98, TA1537)
+S9 mix; 0, 39.1, 78.1, 156, 313, 625, 1250, 2500, 5000 μg/plate(WP2 uvrA)
+S9 mix(additional test, Hamster S9); 0, 1.17, 2.34, 4.69, 9.38, 18.8, 37.5, 75.0, 150, 300 μg/plate(TA98)
　S9:Rat liver, induced with phenobarbital and 5,6-benzoflavone
Hamster liver, not induced with phenobarbital and 5,6-benzoflavone
　Plates/test:3
　Number of replicates:2
GLP:Yes

　Test results:

No increase in revertant colonies was observed in the test with either the non-activation method (-S9) or activation (+S9).

Genetic effects:
Salmonella typhimurium TA100, TA1535, TA98, 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)2)

Purity:99 % and or more
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:1 % Carboxymethyl cellulose sodium salt solution
　Positive controls:-S9 mix; Mitomycin C
+S9 mix; Cyclophosphamide
　Doses:-S9 mix(short-term treatment); 0, 1250, 2500, 5000 μg/mL
+S9 mix(short-term treatment); 0, 1250, 2500, 5000 μg/mL
-S9 mix(continuous treatment 24 hr); 0, 1250, 2500, 5000 μg/mL
　S9:Rat liver, induced with phenobarbital and 5,6-benzoflavone
　Plates/test:2
GLP:Yes

　Test results:

No increase in chromosomal aberrations was observed in the test with either the short-term treatment (-S9 and +S9) or continuous treatment.

Genetic effects:
clastogenicitypolyploidy
+?-+?-
　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, 257-8523, 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