液相色谱-串联质谱法检测动物肝肾组织中3种雌激素

建立了动物肝、肾组织中雌酮、17β-雌二醇、雌三醇药物残留检测的液相色谱-串联质谱(LC-MS/MS)分析方法。样品以叔丁基甲醚提取,于45 ℃下氮吹至干,用正己烷-二氯甲烷(6:4, v/v)重新溶解,过硅胶固相萃取小柱净化,氮吹至干,用1 mL乙腈-水(7:3, v/v)定容后测定,用Poroshell 120 EC-C18柱分离,以乙腈和水作为流动相进行梯度洗脱,电喷雾负离子模式电离,多反应监测模式检测,外标法定量。该方法对3种雌激素的线性范围均为1.0～20.0 μg/kg,相关系数大于0.99,定量限为1.0 μg/kg。在不同基质中,1.0、2.0、10.0 μg/kg 3个添加水平的平均回收率范围为70.2%～114%,相对标准偏差范围为2.01%～14.5%。该方法具有快速简便、净化效果好、灵敏度高等特点,适用于动物肝、肾组织中雌激素的检测。

Determination of three estrogens in animal liver and kidney tissues by liquid chromatography-tandem mass spectrometry

A liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the determination of estrone, 17β-estradiol, estriol in animal liver and kidney tissues. The sample was extracted by tert-butyl methyl ether, and the extract was evaporated by nitrogen at 45 ℃. The residue was redissolved in hexane-dichloromethane (6:4, v/v), then purified on a silica solid-phase extraction column. The eluant was evaporated by nitrogen, dissolved in acetonitrile-water (7:3, v/v) and then analyzed by LC-MS/MS. The separation of estrogens was performed on a Poroshell 120 EC-C18 column with the mobile phases of acetonitrile and water with a gradient elution. The identification and quantification of estrogens were carried out by negative electrospray ionization in the multiple reaction monitoring mode using external standard method. The calibration curves showed good linearity in the range of 1.0~20.0 μg/kg with the correlation coefficients above 0.99. The limit of quantification was 1.0 μg/kg for each estrogen. The average recoveries of the estrogens spiked at 1.0, 2.0, 10.0 μg/kg levels in different matrices were between 70.2% and 114%, and the relative standard deviations were between 2.01% and 14.5%. The method is simple, rapid, sensitive, good in purification effect. It is suitable for the confirmation and quantification of estrogens in animal liver and kidney tissues.