液相色谱-二级质谱联用法测定人血浆中醋酸甲羟孕酮的浓度

 目的建立测定人血浆中醋酸甲羟孕酮的液相色谱-二级质谱联用法(LC-MS/MS)。方法取血浆,加入内标醋酸甲地孕酮,经正己烷液-液萃取后,在50℃水浴以氮气流吹干,残渣用流动相溶解,进行LC-MS/MS测定。色谱柱为Alltech Alltima-C₁₈柱(2.1 mm×100 mm,3μm);以甲醇-0.1%甲酸溶液(72∶28)为流动相;流速0.2 mL·min_-1;柱温40℃;电喷雾离子源(ESI),以选择性正离子方式检测;扫描方式为选择反应监测(SRM),用于定量分析的二级碎片离子m/z分别为327.4(醋酸甲羟孕酮)和325.4(醋酸甲地孕酮,内标)。结果醋酸甲羟孕酮的线性范围为0.10～8.0μg·L^-1,最低检测限为40 ng·L^-1,提取回收率为76.1%,日内、日间精密度(RSD)均小于9.0%。结论本方法专属性强,灵敏度高,操作简便,适用于临床药物检测。

Determination of Medroxyprogesterone Acetate in Human Plasma by LC-MS/MS Spectrometry

OBJECTIVE To develop a sensitive and specific LCMS/MS method for the determination of medroxyprogesterone acetate in human plasma.METHODS Medroxyprogesterone acetate and internal standard megestrol acetate were extracted with liquid-liquid extraction with N-hexane.The organic layer was blowed by nitrogen to dryness at 50 ℃.The residue was reconstituted in the mobile phase.The LC-MS/MS method was performed on an Alltech Alltima-C₁₈ column(2.1 mm×100 mm,3 μm).The mobile phase consisted of methanol and 0.1% methanoic acid(72∶28).The flow rate was at 0.2 mL·min^-1.The temperature of column was at 40 ℃.The compound was ionized in the electrospray ionization(ESI) and operated in positive mode.The scanner mode was selection response monitor(SRM).The second order fragment ion of monitor was at 327.4(medroxyprogesterone acetate)and m/z 325.4(megestrol acetate,internal standard).RESULTS The linear range of the calibration curve for medroxyprogesterone acetate was 0.10～8.0 μg·L^-1.The limit of quantification was 40 ng·L^-1.The relative recovery was 76.1%.The inter and intra-day precision(RSD) were blow 9.0%.CONCLUSION The method is proved to be sensitive,specificity.It is suitable to be applied in clinical drug testing.