Determination of isoniazid in rabbit's vitreous humor and spinal fluid by reversed-phase high performance liquid chromatography / 中国法医学杂志

ABSTRACT

Objective Develop a reversed-phase high performance liquid chromatography (HPLC) methodfor detecting isoniazid in vitreous humor and spinal fluid.Method Vanillin, as a derivative reagent, was added to the vitreous humor and spinal fluid samples. Isoniazid and vanillin reacted to form isonicotinoyl hydrazone which was separated and detected. The pretreatment method of sample, the linear range, the precision, the recovery of isoniazid were all established by using rabbit's vitreous humor and spinal fluid spiked with standard isoniazid. The HPLC method has then been applied to investigate the concentration of isoniazid in intoxicated rabbits'vitreous numor and spinal fluid respectively.Results As established in the method, the linear range was 0.2?g/ml～12.0?g/ml (for vitreous humor ?=0.9990, for spinal fluid ?=0.9988). The detective limit was 0.2?g/ml. The intra and inter-day precision of assay for isoniazid were less than 4.9%( n =5) in vitreous numor and spinal fluid. The average recoveries of isoniazid were more than 97.1%. The concentration of isoniazid was 74.60?7.40?g/ml in vitreous humor, 88.95?10.12?g /ml in spinal fluid.Conclusion The HPLC method is suitable for analyzing isoniazid in the vitreous numor and spinal fluid.