LC-MS/MS测定人血清中奥卡西平及其活性代谢产物10-羟基卡马西平的浓度及其临床应用

Determination of Oxcarbazepine and Its Active Metabolite 10-Hydroxycarbamazepine in Human Serum by LC-MS/MS and Its Clinical Application

关键词:  奥卡西平  10-羟基卡马西平  LC-MS/MS  血药浓度  临床应用

Key words:  oxcarbazepine  10-hydroxycarbamazepine  LC-MS/MS  serum concentration  clinical application

摘要:

目的 建立LC-MS/MS同时测定人血清中奥卡西平（oxcarbazepine，OXC）及其活性代谢产物10-羟基卡马西平（10-hydroxycarbamazepine，MHD）的浓度，并将其应用于临床药物浓度检测。方法 血清样本以卡博替尼为内标，乙腈处理后，用LC-MS/MS进行测定。色谱柱为Waters BEH C₁₈（2.1 mm×150mm，1.7 μm），柱温40℃，流动相为0.1%甲酸和乙腈，流速为0.3 mL·min^-1，进样量1 μL，采用电喷雾正离子多反应监测（MRM）模式检测。OXC及MHD的检测离子对分别为OXC[M-H⁺]m/z 253.2→180.1，MHD[M-H⁺]m/z 255.3→194.2。结果 OXC、MHD血药浓度分别在0.01~1 μg·mL^-1（r²=0.9958），0.4~40 μg·mL^-1（r²=0.996 2）内线性关系良好，准确度在85%~115%，日内、日间精密度RSD均<15%。OXC、MHD提取回收率分别为90.75%~100.43%，80.40%~82.30%。本方法不受基质效应影响，在室温放置、反复冻融、长期保存下都稳定，RSD均<15%。临床检测60例患者血药浓度，OXC中位值为0.48 μg·mL^-1，MHD中位值为17.1 μg·mL^-1。结论 该方法操作简便快速、准确度高、灵敏度强，适用于对OXC及其活性代谢产物（MHD）的临床血药浓度监测，为个体化给药提供检测技术。

Abstract:

OBJECTIVE To establish a LC-MS/MS method for simultaneous determination of oxcarbazepine(OXC) and its active metabolite 10-hydroxycarbamazepine(MHD) in human serum and apply it to the determination of clinical drug concentration. METHODS Serum samples with cabozantinib as internal standard, were determined by LC-MS/MS after acetonitrile treatment. The chromatographic conditions were as following:column was Waters BEH C₁₈(2.1 mm×150 mm, 1.7 µm), temperature was 40℃, mobile phase consisted of 0.1% formic acid and acetonitrile, flow rate was 0.3 mL·min^-1, injection volume was 1 µL. The serum samples was detected by electrospray postive ion source in MRM mode. The detection ion pairs were OXC[M-H⁺]m/z 253.2→180.1, MHD[M-H⁺]m/z 255.3→194.2. RESULTS The serum concentration of OXC and MHD had a good linear relationship in the range of 0.01-1 µg·mL^-1(r²=0.995 8), 0.4-40 µg·mL^-1(r²=0.996 2), respectively. Accuracy ranged from 85% to 115%. The intra-day RSD and inter-day RSD were both < 15%. The extraction recoveries of OXC and MHD were 90.75%-100.43% and 80.40%-82.30%, respectively. The method was not affected by matrix effect. It was stable at room temperature, repeated freeze-thaw and long-term storage, and the RSD was < 15%. The drug concentration of 60 patients were determined by established method, and the median of OXC and MHD concentration were 0.48 and 17.1 µg·mL^-1, respectively. CONCLUSION This method is simple, rapid, accurate and sensitive. It is applicable to the monitoring of the clinical serum concentration of OXC and MHD. It can also provide detection technology for individualized drug therapy.