HPLC法测定人血清中甲砜霉素浓度

Determination of Thiamphenicol Concentration in Human Serum by HPLC

目的:建立测定人血清中甲砜霉素浓度的RP HPLC法。方法:以HypersilODSC18(150mm×4.6mm,5μm)为色谱柱,流动相为乙腈水(25∶75),流速为1.0ml·min-1,检测波长为225nm,柱温35℃,血清样品以乙酸乙酯为提取剂。结果:甲砜霉素在0.05～20.00mg·L-1浓度范围内线性关系良好(r=0.9992),高、中、低3个浓度的平均回收率分别为98.05%,100.10%,100.30%;最低检测浓度为0.01mg·L-1。结论:该方法精密、准确,可用于甲砜霉素的药动学研究。

Objective: To establish a method for determination of the concentration of thiamphenicol in human serum by the RP-HPLC.Method: A Hypersil ODS C_(18) column was used with a mobile phase of acetonitrile-water(25∶75),the flow rate was (1.0) ml·min^(-1),the detecting wavelength was at 225 nm, the column temperature was (35 ℃) . Serum sample((0.5) ml) was extracted with (2.5) ml of ethyl acetate in a rotary mixer,the organic phase was completely evaporated under a nitrogen stream in a water bath at (40 ℃).The residue was redissolved in 200 μl mobile phase.Result: The calibration curve was a good linear, within a concentration range of (0.05)～(20.00) mg·L^(-1), r=(0.999 2)(n=11);The average recoveries concentrations of three different thiamphenicol were (98.05%),(100.10%), and (100.30% ), respectively;The limit of quantitation for thiamphenicol was (0.01) mg·L^(-1). Conclusion: The method is accurate and precise,It is usable for the study of clinical thiamphenicol pharmacokinetics.