固相萃取-超高效液相色谱-串联质谱法同时测定饲料中的3种雌激素

Simultaneous determination of three estrogens in feed by solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry

建立了固相萃取-超高效液相色谱-串联质谱（ SPE-UPLC-MS / MS）同时测定饲料（预混合、配合和浓缩饲料）中3种雌激素（17β-雌二醇、苯甲酸雌二醇和戊酸雌二醇）的检测方法。饲料样品经乙腈提取，Heaion C 18固相萃取柱净化后，用 ACQUITY UPLC BEH C18色谱柱（50 mm×2.1 mm，1.7μm）分离，乙腈和0.1％氨水溶液作为流动相进行梯度洗脱，多反应监测（MRM）模式检测，内标法定量。结果表明，17β-雌二醇和戊酸雌二醇在10-200μg / L、苯甲酸雌二醇在5-200μg / L 范围内具有良好线性，相关系数（r）≥0.996。17β-雌二醇、苯甲酸雌二醇和戊酸雌二醇的检出限分别为7、5和7μg / kg。样品的平均加标回收率为96.5％-102.0％。该法前处理操作简便、分析速度快、检出限低、准确度高，可用于饲料中3种雌激素的同时检测。

A method was developed for the simultaneous determination of three estrogens （17β-estradiol,estradiol benzoate,estradiol valerate）in feed by solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry（SPE-UPLC-MS / MS）. After the sample was extracted by acetonitrile and cleaned-up on a Heaion C 18 solid phase extraction （ SPE）column,the three estrogens were separated by an ACQUITY UPLC BEH C 18 column（50 mm×2. 1 mm,1. 7 μm）with gradient elution using acetonitrile and 0. 1% ammonia solution as mobile phases and finally confirmed and quantified in multiple reaction monitoring（ MRM ） mode. The internal standard calibration curves were used for the quantification. The results showed that all the estrogens were determined with excellent linear relationships from 10 μg / L to 200 μg / L for 17β-estradiol and estradiol valerate,and from 5 μg / L to 200 μg / L for estradiol benzoate. The correlation coefficients（ r）of the three estrogens were not lower than 0. 996. The limits of detection for 17β-estradiol, estradiol benzoate and estradiol valerate were 7μg / kg,5 μg / kg and 7 μg / kg,respectively. The average recoveries were 96. 5% - 102. 0% . The results demonstrated that the developed method can simultaneously determine the three estro-gens in feed with advantages of simple pretreatment,rapid analysis,low limit of detection and high accuracy.