HPLC法测定宫外孕患者血清中甲氨蝶呤的浓度

Determination of serum concentration of methotrexate in patients with ectopic pregnancy by HPLC

目的建立高效液相色谱法实时监测宫外孕患者血清中甲氨蝶呤浓度。方法以Zorbax SB-C18柱(4.6×250mm,5μm)为色谱柱,流动相为0.05mol·L-1磷酸盐缓冲液(pH6.8)-甲醇(78∶22),流速1.0mL·min-1,检测波长306nm。以高氯酸为蛋白沉淀剂。结果甲氨蝶呤在0.61～19.52μg·mL-1内线性关系良好,r=0.9997(n=5),检测限为0.15μg·mL-1。高、中、低3浓度的方法回收率为97.54%～99.80%,提取回收率为81.15%～85.47%。日内和日间RSD均小于7%。结论该方法简便、准确、重复性好,适用于临床甲氨蝶呤血药浓度监测。

OBJECTIVE To establish a HPLC method for the determination of MTX in human serum. METHODS Separation and determination were achieved on a Zorbax SB-C18 column （4.6 ×250mm,5μm） with the mobile phase consisted of 0.05mol· L^- 1： phosphate buffer（pH6.8） methanol（78 ： 22） at a flow rate of 1.0mL· min^-1. The UV detection wavelength was set at 306nm. The serum samples were treated with perchloric acid for sedimentation. RESULTS The good linearity was from 0.61 to 19.52μg·mL^-1,r= 0.9997（n= 5） .The minimum detectable concentration of methotrexate was 0.15μg·mL^-1 .The method recoveries of methotrexate in high, medium and low concentrations were from 97.54 % to 99.80 %, and the extraction recover-ies were from 81.15 % to 85.47 %. The within - day and between - day precision of variation was less than 7 %. CONCLUSION The method is simple, accurate and reliable. It is suitable for the blood drug concentration monitoring of methotrexate in the clinic.