高效液相色谱法测定人血浆中拉莫三嗪、奥卡西平及其代谢物浓度

Determination of concentrations of lamotrigine, oxcarbazepine and its active metabolite in human serum by HPLC method

目的：建立测定人血浆中拉莫三嗪、奥卡西平及其代谢物10－羟基卡马西平浓度的高效液相色谱方法。方法以非那西汀为内标，血浆样品用乙醚萃取，用C18柱分析，流动相为乙腈－0．05 mol· L－1磷酸二氢钠水溶液（25∶75， pH 4．5），流速为1．0 mL· min－1，柱温为25℃，检测波长为220 nm。同时考察该方法的专属性、标准曲线和定量下限、精密度与回收率、稳定性。结果拉莫三嗪、10－羟基卡马西平、奥卡西平分别在0．1～20．0μg· mL－1（y＝20．11×10^－2c＋1．31×10^－2，r ＝0．9990），0．5～50．0μg· mL－1（y ＝9．77×10^－2 c ＋0．35×10^－2， r＝0．9998），0．25～25．0μg· mL－1（ y ＝131．69×10^－2 c －0．59×10^－2， r＝0．9991）线性关系良好。拉莫三嗪、10－羟基卡马西平、奥卡西平的方法回收率均在88．65％～100．38％，日内、日间精密度均＜15％。结论该方法操作简便、准确、灵敏度高，可用于拉莫三嗪、奥卡西平及其代谢物10－羟基卡马西平血药浓度监测。

Objective To establish an HPLC method to determinate the concentrations of lamotrigine , oxcarbazepine and its active metabolite simultaneously in human serum. Methods Phenacetin was used as internal standard, and the plasma was extracted by diethyl ether.The HPLC method was performed on the C18 column. The mobile phase consisted of acetonitrile -0.05 mol· L-1 biphosphate sodium （ 25∶75, pH 4.5）.The flow rate was 1.0 mL· min -1 and the column temperature was kept at 25 ℃.The detecting wavelength was set at 220 nm.The method was inspected in the aspects of specificity, lower limit of quantifi-cation, precision, recovery rate and stability.Results The linear range of lamotrigine, monohydroxycarbazepin and oxcabazepine were from 0.1 to 20.0 μg· mL-1 （ y=20.11 ×10^-2 c ＋1.31 ×10^-2 , r =0.999 0 ） , 0.5-50.0 μg· mL-1 （ y=9.77 ×10^-2 c＋0.35 ×10^-2 , r=0.999 8 ） , 0.25 -25.0 μg· mL-1 （ y =131.69 ×10^-2 c -0.59 ×10^-2 , r=0.999 1） , respectively.The range of extraction recovery ratios were both between 88.65%and 100.38%.The intra and inter-day precision were both less than 15%.Conclusion The method is shown to be sim-ple, accurate and sensitive, which can be applied in the determination of concentration of lamotrigine, oxcarbazepine and its active metabolite in human plasma.