Genotoxicity evaluation of 4-carboxyl- 2,6-dinitrophenylazohydroxynaphthalenes in mice.

A short-term in vivo genotoxicity evaluation of 4-carboxyl-2,6-dinitrophenylazohydronaphthalenes (AZ-01 to AZ-04) has been carried out in mice. Aqueous colloidal solutions of the dyes were administered to mice on each day for 5 successive days using gastric gavages. Two end point assessments of the genotoxicity potentials of the dyes were assessed using comet assay and chromosomal aberration studies using the mice bone marrow cells. The dyes were well tolerated at the doses investigated, as there were no deaths or any adverse pharmacotoxic events. Dose-dependent DNA damage (in terms of percentage of tail DNA and Olive tail moment) occurred with AZ-01 and AZ-02, although the effects were significant only with the highest doses. AZ-03 gave similar patterns with those of AZ-01 and AZ-02, while replacement with butanone in AZ-04 altered the observed pattern. Minimal chromosomal damages were obtained for the four dyes, with AZ-01 and AZ-02 giving nonsignificant damages, while the highest dose of AZ-03 produced significant aberrations in terms of breaks. Some minor isochromatid breaks and gaps were also noticed in the dye-treated mice. Mitotic indices in all cases were not significantly different from concomitantly administered vehicle control showing lack of cytotoxicity of the monoazo dyes at these doses. The monoazo dyes show the potential of being utilized as colorants, pending further required tests.

A new approach to the spectrophotometric determination of metronidazole and tinidazole using p-dimethylaminobenzaldehyde.

A new approach to the spectrophotometric determination of metronidazole (MZ) and tinidazole (TZ) has been developed. The procedure involves coupling of diazotized nitroimidazoles with p-dimethylaminobenzaldehyde (DMAB) to form a greenish-yellow solution. Optimal temperature and time were 0 degrees C (iced) and 3 minutes for diazotization and 30 degrees C and 2 minutes for coupling for both MZ and TZ. Coloured adducts of MZ and TZ showed shoulders at 406 nm and 404 nm, respectively, which were selected as analytical wavelengths. The reaction with p-DMAB occurred in a 1:1 mole ratio. Beer's law was obeyed within the 4.8-76.8 microg mL(-1) concentration range with low limits of detection. The azo adducts were stable for over a week. Molar absorptivities were 1.10 x 10(3) (MZ) and 1.30 x 10(3) L mol(-1) cm(-1) (TZ). Overall recoveries of MZ and TZ from quality control samples were 103.2 + or - 1.3 and 101.9 + or - 1.3% over three days. There was no interference from commonly utilized tablet excipients. No significant difference was obtained between the results of the new method and the BP titrimetric procedures. The azo approach using the p-dimethylaminobenzaldehyde procedure described in this paper is simple, fast, accurate and precise. It is the first application of DMAB as a coupling component in the diazo coupling reaction.

Novel colorimetric assay of indomethacin using 4-carboxyl-2,6-dinitrobenzene diazonium ion.

A simple, sensitive and direct colorimetric method for the determination of indomethacin either in pure form or in capsules has been developed. The method is based on the diazo coupling reaction between indomethacin and a highly reactive arenediazonium ion, 4-carboxyl-2,6-dinitrobenzene diazonium ion, with the consequent formation of an azo dye. The reaction is fast and gave an orange azo dye in ethyl acetate. The assay was carried out at 470 nm and the azo adduct was stable for three hours. Beer's law is obeyed in the concentration range of 3.3-11 microg mL(-1) of indomethacin. Optimization studies established an optimum reaction time of 20 minutes at 30 degrees C and the drug-to-reagent ratio of 1:2 for optimal detector response. The method developed has a low limit of detection of 0.90 microg mL(-1) and is precise (RSD 2.3%). The new method has been successfully applied to the determination of indomethacin in capsules and the method is of equivalent accuracy as the official (BP) spectrophotometric method. The new method could find application as a simple analytical method for the assay of indomethacin in capsules.