| Summary: | High-performance liquid chromatography is one of the most frequently used separation methods for an analysis of drugs in biological material. It is suitable both for quantitative and qualitative drug analysis. Isoniazid is used for the treatment and a prevention of tuberculosis. Acute isoniazid intoxication is manifested by tonic-clonic convulsions. Vitamine B6 (pyridoxine) is used in clinical practice as an antidotum. Although this type of toxicity is considered to be associated with the deficiency of vitamine B6 (or its active form, pyridoxal-5-phosphate - PLP), description of mechanisms and processes, which are responsible for this effect, is still missing. The aim of this study was to develop suitable chromatographic conditions for monitoring intoxication by isoniazid and detection of potential hydrazone metabolites. These conditions should allow a separation of seven analytes (isoniazid, acetylisoniazid, pyridoxal, pyridoxol, pyridoxal-5-phosphate, and the o-108 - internal standard). Furthermore, the aim was to study possibilities of isolating all those substances from rabbit plasma. The best chromatographic separation was achieved using the HPLC column LiChroCART 250 × 4 nm I.D., packed with Lichrospher 100 RP-18 (5 µm). The mobile phase was composed of a buffer - component A (NaH2PO4 0.01 mol/l, with...
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