家兔玻璃体和脑脊液中异烟肼的高效液相色谱分析

Determination of isoniazid in rabbit's vitreous humor and spinal fluid by reversed-phase high performance liquid chromatography

目的 建立玻璃体和脑脊液中异烟肼的高效液相色谱分析方法。方法 以香草醛为衍生化试剂 ,经柱前衍生为异烟肼 -香草醛腙 ,直接对体液样品中的腙进行定性、定量分析 ,并以空白兔体液添加标准异烟肼对样品的前处理方法、仪器条件、线性范围、精密度、回收率进行全面考查后 ,测定急性染毒家兔玻璃体和脑脊液中的异烟肼浓度。结果 用上述方法分析兔玻璃体和脑脊液中异烟肼 ,其线性范围为 0 2～ 12 0 μg/ml,检测限 0 2 μg/ml,日内、日间精度均小于 4 9% ,回收率均在 97 1%以上。异烟肼在急性染毒家兔玻璃体中的浓度为 74 60± 7 40 μg/ml;脑脊液中为 88 95± 10 12 μg/ml。 结论 本文所建方法适用于异烟肼中毒者玻璃体和脑脊液中异烟肼的检测。

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 0.2μg/ml～12.0μg/ml (for vitreous humor γ=0.9990, for spinal fluid γ=0.9988). The detective limit was 0.2μg/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±7.40μg/ml in vitreous humor, 88.95±10.12μg /ml in spinal fluid.Conclusion The HPLC method is suitable for analyzing isoniazid in the vitreous numor and spinal fluid.