Determination of Isoniazid Gained Fluorescent with 9-Anthraldehyde by HPLC-FL Method in Dosage Forms

Abstract

Tuberculosis is a chronic necrotizing bacterial infection caused by Mycobacterium tuberculosis, which has not lost its prevalence in the society from the past to the present. Isoniazid (INH) is an important drug used in the treatment of tuberculosis and is used for all forms of tuberculosis caused by Mycobacterium tuberculosis, unless there is resistance. This study is aimed to develop and validate a new HPLC with fluorescence detector (HPLC-FL) based on the formation of a coloured product (9-anthraldehyde isonicotinoyl hydrazone, INHAA) resulting from the condensation reaction of INH with 9-anthraldehyde(9-AA) for the determination of INH in dosage forms. Optimum INHAA formation was obtained at 68 °C and 1.97 mL 0.05 M 9-AA for 3 h using a chemometric tool which is one of the green analytical chemistry strategies. INHAA was separated on a C18 reversed-phase column in conjunction with an isocratic elution of a mobile phase composed of methanol-Milli-Q grade water and detected by fluorescence detection (FL) at excitation (390 nm) and emission (489 nm) wavelengths. The method was linear over concentration range that varies between 0.5 and 20 µg/mL INH. The limit of detection (LOD) and limit of quantitation (LOQ) were estimated based on three and ten times the baseline noise and were found to be 0.11 µg/mL and 0.33 µg/mL INH, respectively. The optimized HPLC-FL parameters were applied to the analysis of drug dosage forms INH containing for quality control purposes. A new HPLC-FL method was developed for the analysis of INH in drug dosage forms, which saves time, effort and chemicals in the experimental design.