Solid-Phase Extraction and HPTLC Determination of Isoniazid and Acetylisoniazid in Serum. Comparison with HPLC

A new solid-phase extraction (SPE) and quantitative determination of isoniazid (INH) and its acetyl metabolite (AcINH) in serum by high-performance thin-layer chromatography (HPTLC) is presented. Alkalized serum samples with nicotinamide as an internal standard are applied to an SPE cartridge containing a new SPE sorbent, [poly ( d i v i n y l b e n z e n e - c o -N-vinylpyrrolidone )]. A simple procedure of conditioning, washing, and eluting steps is described. After evaporation of the eluates to dryness and reconstitution, onedimensional HPTLC is performed on silica gel plates with ethyl acetate‐methanol (70:30) as a mobile phase. Quantitation is done by densitometry. Convenient validation parameters are obtained ( l i n e a r i t y, limits of detection and quantitation, precision, and accuracy) for INH and AcINH. The method is compared with a high-performance liquid chromatographic (HPLC) technique developed in the laboratory, and satisfactory correlation is found between data from the two techniques. The HPTLC method is sensitive and specific and is used to quantitate INH and AcINH in patient blood serum, and the results are compared with those obtained by HPLC.