反相高效液相色谱法测定人血浆中丙氧咪唑浓度

Determination of oxibendazole in human plasma by reversed phase of HPLC

目的建立测定人血浆中丙氧咪唑浓度的高效液相色谱法。方法血浆上样于C18SPE柱后,先用15%甲醇冲洗,再用90%甲醇洗脱,N2气流下挥干。采用反相高效液相色谱法,紫外检测器检测。色谱柱为ODS柱,流动相为含1.25%三乙胺溶液∶甲醇∶乙腈(70∶15∶15,V/V/V),流速为1.0 ml.min-1,检测波长(λ)295 nm。结果本方法在20-600μg.L-1范围内线性良好(r=0.998 6,n=6)。高、中、低浓度的相对回收率均在92.0%-108.2%之间,日内与日间RSD均小于8.9%,最低检测浓度为5μg.L-1。结论本法适用于人血浆中丙氧咪唑浓度的测定。

Aim To establish a high-performance liquid chromatographic method for the determination of oxibendazole in human plasma. Methods The oxibendazole were extracted fi＇om human plasma by loaded onto a conditioned C18 SPE column, rinsed with 15% methanol, and eluted with 90% methanol. After extraction, the separation of oxibendazole from internal standard （albendazole） and endogenous components was accomplished by using reversed phase chromatography. The mobile phase, a combination of 1.25 % triethylamine in water, methanol and acetonitrile （ 70：15： 15, V/V/V） , was run through a Ultrasphere-ODS （ 250 mm × 4.6 mm, 5 μm ） analytical column. The flow rate was 1.0 ml· min^-1. The UV detection wavelength was at 295 nm. Results The assay exhibited excellent linear relationships （ r = 0.9986） between peak area and plasma concentrations over the range of 20 - 600μg.L^-1 ; quantitation limit was at least 5 μg.L^-1,and mean recovery was between 92.0% and 108.2%. Accuracy and precision were below 8.9% both interday and intraday. Conclusion The method is suitable for the measurement of oxibendazole in human plasma.