Absorption, Distribution and Excretion
MORE THAN HALF OF BOUND DYE IN ALL LIVER-CELL FRACTIONS IS ASSOC WITH SOL PROTEINS; OF THIS, 80% IS BOUND TO FRACTION WHICH ACCOUNTS FOR ONLY 15% OF SOL PROTEINS.
BILIARY EXCRETION OF THE AZO DYE CARCINOGEN, N,N-DIMETHYL-4-AMINOAZO(C-14)BENZENE (DAB) WAS STUDIED IN MALE RATS. AFTER A SINGLE IV DOSE OF CARCINOGEN, 20-25% OF THE INJECTED RADIOACTIVITY APPEARS IN THE BILE WITHIN 1 HR, ENTIRELY AS METABOLITES.
IT WAS ADMIN IP TO WISTAR RATS & FOUND TO HAVE THE TENDENCY TO BIND TO LIVER TISSUES, NUCLEI & CHROMATIN WITH SPECIFIC BINDING AFFINITY TO NUCLEAR NONHISTONE PROTEIN.

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Metabolism / Metabolites
YIELDS 4'-DIMETHYLAMINO-4-HYDROXYAZOBENZENE & 4-METHYLAMINOAZOBENZENE IN RAT. /FROM TABLE/
DAB IS METABOLIZED VIA FOLLOWING PATHWAYS: 1) REDUCTION & CLEAVAGE OF AZO GROUP; 2) DEMETHYLATION; 3) RING HYDROXYLATION; 4) N-HYDROXYLATION; 5) N-ACETYLATION & O-CONJUGATION OF METABOLITES.
IN RATS & MICE, CARCINOGENIC AROMATIC AZO-DYES UNDERGO N-HYDROXYLATION, & FOR EXAMPLE, N-HYDROXY-N-ACETYL-4-AMINOAZOBENZENE IS URINARY METABOLITE OF HEPATOCARCINOGENIC 4-DIMETHYLAMINOAZOBENZENE ... .
A study was made of the azoreduction of dimethylaminoazobenzene (DAB) in-vitro by purified rat liver NADPH cytochrome p450-reductase and by a partially purified reconstituted cytochrome p450 system. Male Wistar rats were induced with clofibrate, nafenopin, phenobarbital, or beta-naphthoflavone prior to liver microsomal isolation. ... Both purified NADPH cytochrome p450 reductase and the reconstituted system reduced dimethylaminoazobenzene, with a more than five fold greater rate for the latter. A ratio of 2:1 azoreductase to cytochrome p450 was optimal for dimethylaminoazobenzene reduction. Clofibrate was the only inducer of azoreductase. Reaction with the suicide inhibitor 10-undecynoic acid distinguished laurate hydroxylase from the azoreductase. The reconstituted system was active in air and carbon monoxide, while purified NADPH cytochrome p450 reductase was inhibited in air. In both systems, nearly 50 fold acceleration of dimethylaminoazobenzene azoreduction occurred in the presence of flavin adenine dinucleotide or flavin mononucleotide, but only under anaerobic conditions, indicating facilitation of electron flow to dimethylaminoazobenzene onlhy from NADPH cytochrome p450-reductase and not from cytochrome p450 itself. This was confirmed with the use of cytochrome p450 inhibitors. ... There appears to be an obilgatory electron flow through cytochrome p450 to dimethylaminozaobenzene, bypassing an oxygen sensitive step, and dietary riboflavin probably attenuates dimethylaminoazobenzene hepatocarcinogenicity solely by its effect on intestinal bacteria.
For more Metabolism/Metabolites (Complete) data for 4-(DIMETHYLAMINO)AZOBENZENE (7 total), please visit the HSDB record page.

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Biological Half-Life
No reports found; [TDR, p. 547]

[1]. Nakano N, et, al. An automatic monitor of formaldehyde in air by a monitoring tape method. J Environ Monit. 1999 Jun;1(3):255-8.

4-Dimethylaminoazobenzene can cause cancer according to an independent committee of scientific and health experts.
4-dimethylaminoazobenzene appears as yellow crystalline leaflets or an orange powder. (NTP, 1992)
4-(dimethylamino)azobenzene is a member of azobenzenes.
4-Dimethylaminoazobenzene is used as a dye for coloring polishes, wax products, and soap. Acute (short- term) dermal exposure to 4-dimethylaminoazobenzene may result in contact dermatitis in humans. No information is available on the chronic (long-term), reproductive, developmental, or carcinogenic effects of 4-dimethylaminoazobenzene in humans. Animal studies have reported birth defects in the offspring of mice exposed to 4-dimethylaminoazobenzene and tumors of the lung, liver, and bladder from oral exposure to 4-dimethylaminoazobenzene. EPA has not classified 4-dimethylaminoazobenzene for carcinogenicity. The International Agency for Research on Cancer (IARC) has classified 4- dimethylaminoazobenzene as a Group 2B, possibly carcinogenic to humans.
4-Dimethylaminoazobenzene is a yellow, crystalline solid compound. 4-Dimethylaminoazobenzene was used as a dye for coloring polishes, wax products, polystyrene and soap, and was also used as a pH indicator, but is no longer used or produced in the United States. Exposure to this substance causes dermatitis. 4-dimethylaminoazobenzene is reasonably anticipated to be a human carcinogen. (NCI05)
A reagent used mainly to induce experimental liver cancer. According to the Fourth Annual Report on Carcinogens (NTP 85-002, p. 89) published in 1985, this compound may reasonably be anticipated to be a carcinogen. (Merck, 11th ed)

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Mechanism of Action
4-Dimethylaminoazobenzene (or butter yellow) has been used as a laboratory hepatocarcinogen to study the mode of action of this class of compounds, including structure activity relationships. Because of its characteristic color change as a function of pH, it was the first carcinogen that demonstrated binding to cellular macromolecules. Because of the flavin-dependent azo dye reductase, it was also one of the first carcinogens with a clear-cut nutritional modulation of its activity. Substitution with polar groups such as sulfonic acid in simple azo dyes usually abolishes carcinogenicity and mutagenicity. Thus, molecules such as FD and C Red No. 2 (amaranth) are not mutagenic or carcinogenic. On the other hand, tetra-azo dyes such as direct black 38 or direct blue 6, which are not mutagenic and are quite polar, can release the mutagenic carcinogenic benzidine upon reduction of the azo link by bacteria. Such dyes are highly carcinogenic.

C14H15N3

225.2890

225.126

60-11-7

N,N-dimethyl-4-(phenyldiazenyl)aniline-d5;1398109-08-4

6053

YELLOW CRYSTALLINE LEAFLETS
Yellow, leaf-shaped crystals.

1.0±0.1 g/cm3

371.0±25.0 °C at 760 mmHg

111 °C (dec.)(lit.)

178.2±23.2 °C

0.0±0.8 mmHg at 25°C

1.567

4.43

0

3

3

17

236

0

N(C([H])([H])[H])(C([H])([H])[H])C1C([H])=C([H])C(=C([H])C=1[H])/N=N/C1C([H])=C([H])C([H])=C([H])C=1[H]

JCYPECIVGRXBMO-UHFFFAOYSA-N

InChI=1S/C14H15N3/c1-17(2)14-10-8-13(9-11-14)16-15-12-6-4-3-5-7-12/h3-11H,1-2H3

N,N-dimethyl-4-phenyldiazenylaniline

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)