【摘要】 目的 建立一种人体血清中心脏毒素的超高效液相色谱串联质谱（UPLC-MS-MS）的检测新方法。方法 向100μl血清样品中加入300μl乙腈沉淀蛋白，-20℃冷冻2 h，高速离心10 min，取上层有机相用纯水按照1∶1比例稀释，然后过膜、上机测定。样品分离采用梯度洗脱模式，流动相为0.1%甲酸水溶液-0.1%甲酸乙腈溶液，柱温为70℃，Protein BEH C4色谱柱分离，采用电喷雾离子源（ESI）正离子模式测定血清中心脏毒素浓度，外标法定量分析。结果 心脏毒素在2.0～200.0μg/L范围内呈现良好的线性关系，r=0.999 2。该方法的回收率为98.6%～115.0%,RSD为3.6%～9.8%；检出限为0.6μg/L。结论 该方法操作简单，灵敏度和准确度均较高，适用于血清中心脏毒素的快速测定。更多还原

【Abstract】 Objective To develop a new method for the determination of cardiotoxin in serum by ultra-high performance liquid chromatography tandem with mass spectrometry(UPLC-MS-MS). Methods Totally 100 μl serum sample was precipitated with300 μl acetonitrile for 2 h at-20 ℃ and then separated by high speed centrifugation for 10 min. The upper organic phase was diluted at 1∶1 by pure water,then passed through the membrane,and measured it by UPLC-MS-MS. Separation of the sample was achieved by a Protein BEH C4 column with a gradient elution of 0.1% formic acid aqueous solution-0.1% formic acid acetonitrile solution. The column temperature was 70 ℃. The concentration of cardiotoxin was measured by electrospray ion source(ESI) with positive ion mode and quantified by external standard method. Results This method showed a good linearity for cardiotoxin in the range of 2.0-200.0 μg/L,r=0.999 2. The recovery was in the range of 98.6%-115.0% with RSDs of 3.6%-9.8%. The limit of detection was 0.6 μg/L. Conclusion This method is simple to operate and has high sensitivity and accuracy,and is applicable to rapid detection of cardiotoxin in the serum.更多还原