Ion interactionreagent reversed-phase high-performance liquid chromatography determination of anti-buberculosis drugs and metabolites in biological fluids

摘要

New methods of ion interaction reagent (IIR) RP-HPLC are presented for the determination of anti-tuberculosis drugs and their metabolites, singly or in multi-component mixtures, in biological fluids. The following analytes are considered:isoniazid, ethionamide, pyrazinamide, morphazinamide, p-aminosalicylic acid, nicotinic and isonicotinic acids. Aqueous solutions of three different ion interaction reagents are alternatively or comparatively used as the mobile phases, namely; (A) 5.00 mM octylaniine at pH 3.00 for o-phosphoric acid, (B) 5.00 mM octylaniine at pH 8.00 for a-phosphoric acid, and (C)5.00 mM 1,6 diaminohexane at pH 6.00 for o-phosphoric acid. The response linearity between peak area and analyte concentration is verified for all the analytes in the concentration range within the determination limits and 2.00 mg/I. Detection limits are always lower than 82 ~g/l for standard solutions; in the analysis of samples of rat serum, rat plasma and human serum, the matrix effect is negligible, the detection limits are always lower than 94 ~.agll and the average recovery.yield is always greater than 96%.