Detection and Assay of Isoniazid Utilizing Isocratic High-Performance Liquid Chromatography

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Nguyen, Cassandra
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To demonstrate an analysis for isoniazid from aqueous mixtures and solid tablets, utilizing isocratic conditions, with high performance liquid chromatography (HPLC) and UV detection. Isoniazid in solid and liquid samples is assayed utilizing HPLC. Isoniazid is a tuberculostat, that is assayed by high performance liquid chromatography that utilizes a reversed-phase C-18 column. The eluent solvent is 5.3% ethanol, 93.7% water, and 1% acetic acid. Aqueous samples are prepared in distilled water as the solvent. The ultraviolet-visible detection of isoniazid is accomplished at 265 nm, which is the wavelength having the highest sensitivity for isoniazid. Having a column pump pressure at 2100 psig, rise time 0.1, and with flow rate 1.0 mL/minute, the elution of isoniazid occurred at 2.2 minutes. The standard curve that was applied ranged from zero to 0.001460 molar in concentration (y = 50,237,766.7435 x). The model described by the standard curve had a coefficient of determination of R2 = 0.9921, and a very strong positive correlation coefficient (r= 0.9960). Percent recovery of drug ranged from 97.3% to 105%. Utilizing reversed phase column with isocratic solvent conditions is shown here to be an effective methodology for determination of isoniazid.