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目的建立高效液相荧光法测定人血浆中他莫昔芬及主要代谢物浓度。方法血浆样品经正己烷-正丁醇(98∶2)提取后,以甲醇-1%三乙胺水溶液(82∶18)为流动相,流速为1mL·min-1,色谱柱为Agilent Extend C18(4.6mm×150mm,5μm),柱温为50℃,离线紫外照射(254nm)10.5min,使目标化合物转变成强荧光性的菲衍生物,荧光检测波长:激发波长(λex)260nm,发射波长(λem)375nm。结果血浆内源性杂质不干扰待测物测定,线性范围分别为:他莫昔芬为0.5~200μg·L-1;N-去甲他莫昔芬为0.5~300μg·L-1;4-羟基他莫昔芬为0.1~10μg·L-1。日内、日间精密度(RSD)均小于10%。样品4次冻融,以及在提取后,4℃下12h内稳定性良好。结论该法灵敏、快速、准确,操作简便、线性范围宽,可用于他莫昔芬的药动学研究及常规治疗药物监测。
Objective To establish a HPLC method for the determination of tamoxifen and its metabolites in human plasma. Methods The plasma samples were extracted with n-butanol (98: 2) and methanol-1% triethylamine (82:18) at a flow rate of 1 mL · min-1. The column was Agilent Extend C18 (4.6mm × 150mm, 5μm), the column temperature was 50 ℃, UV irradiation (254nm) for 10.5min, the target compound was transformed into a strong fluorescent phenanthrene derivative. Fluorescence detection wavelength: 260nm excitation wavelength (λex) (λem) 375 nm. Results The plasma endogenous impurities did not interfere with the determination of analytes. The linear range was 0.5 ~ 200μg · L -1 for tamoxifen, 0.5 ~ 300μg · L -1 for n-desmethyl tamoxifen, Hydroxyl tamoxifen was 0.1 ~ 10μg · L-1. Day, day precision (RSD) are less than 10%. Samples were freeze-thawed four times and the stability was good within 12 h at 4 ° C after extraction. Conclusion The method is sensitive, rapid, accurate, easy to operate and has a wide linear range. It can be used to study the pharmacokinetics of tamoxifen and the monitoring of conventional therapeutic drugs.