高效液相色谱-质谱法测定尿液中痕量雷公藤春碱

HPLC-MS Determination of Trace Amount of Wilfortrine in Urine

提出了尿液中雷公藤春碱的高效液相色谱-质谱测定方法。尿液样品经Waters Oasis（MCX固相萃取小柱富集、净化后,以Zorbax Eclipse SB C18反相色谱柱（4.6 mm×250 mm,5μm）为分离柱,以乙酸盐缓冲溶液-乙腈（40＋60）为流动相,采用正离子模式大气压化学电离源,在选择离子监测模式下进行检测,雷公藤春碱的定量离子质荷比（m/z）为874.4。线性范围为0.2~50.0μg·L-1,检出限（3S/N）为0.07μg·L-1,测定下限（10S/N）为0.2μg·L-1。回收率在86.0%~92.0%之间,日内（n=6）和日间（n=15）相对标准偏差分别小于7.5%和10.5%。

A method for the determination of wilfortrine in urine by HPLC-MS was proposed.Wilfortrine in sample was separated,enriched and purified on the Waters Oasis MCX SPE column.The eluate was used for HPLC-MS determination,in which the Zorbax Eclipse SB C18 column（4.6 mm×250 mm,5 μm） was used as chromatographic column,and a mixture of acetate buffer solution and acetonitrile（40＋60） was used as mobile phase.Atmospheric pressure chemical ionization source with positive ionization mode as well as selected ion monitoring mode was used in the detection.Ion used for quantification was m/z 874.4.Linearity range was found between 0.2-50.0 μg·L-1,and values of detection limit（3S/N） and lower limit of determination（10S/N） found were 0.07 μg·L-1 and 0.2 μg·L-1 respectively.Values of recovery were ranged from 86.0% to 92.0%.Values of RSD′s（intra-day,n=6） and RSD′s（inter-day,n=15） were less than 7.5% and 10.5%,respectively.