UPLC-MS/MS法测定美容养颜类保健食品中9种雌激素

Determination of 9 Estrogens in Beautifying Health Products by Ultra Performance Liquid Chromatography Tandem Mass Spectrometry

建立一种快速检测美容养颜类保健食品中非法添加9种雌激素类药物(雌三醇、己烯雌酚、己烷雌酚、己二烯雌酚、17α-炔雌醇、雌酮、17α-雌二醇、17β-雌二醇和己烯雌酚-D8)的UPLC-MS/MS方法。样品经水-甲醇(50:50,V/V)超声提取后,上清液用Oasis HLB固相萃取柱净化,以0.05%的氨水溶液-甲醇为流动相梯度洗脱,经Inertsil ODS-3(2.1 mm×75 mm,2μm)色谱柱分离,目标化合物在ESI源负离子模式下电离,采用多反应监测模式进行定性、定量分析。结果表明:9种雌激素的方法检出限(S/N=3)为0.4~1.3μg/kg,定量限(S/N=10)为1.9~5.4μg/kg。在低、中、高3个加标水平下的平均回收率为76.9%~109.9%,相对标准偏差(N=6)为1.3%~14.7%。对25批次市售美容养颜类保健食品进行检测,均未发现上述9种雌激素。该方法灵敏高、专属性强,适用于美容养颜类保健食品中雌激素类非法添加的定量及确证分析。

To establish an UPLC-MS/MS method for rapid determination of 9 estrogens（estriol, diethylstilbestrol, hexestrol,dienestrol, 17α-ethinylestradiol, estrone, 17α-estradiol, 17β-estradiol and diethylstilbestrol-D8） added in beautifying health products. The samples were extracted with methanol-Water（50：50, V/V） by ultrasonication, and the upper solution was cleaned up with an Oasis HLB solid phase extraction cartridge. After chromatographic separation on an Inertsil ODS-3（2.1 mm×75 mm, 2 μm） column with 0.05% ammonia-methanol as mobile phase, the target compounds were qualitatively and quantitatively determined under the multiple reaction monitoring（MRM） mode in negative ionization mode. The results showed that the limits of detection（LOD） for 9 estrogens were in the range of 0.4-1.3 μg/kg and the limits of quantification（LOQ） were in the range of 1.9-5.4 μg/kg. The average recoveries at three spiked levels ranged from 76.9% to 109.9%, with relative standard deviations（N=6） of 1.3%-14.7%. The method was sensitive, sufficiently selective and was suitable for the determination and confirmation of 9 estrogens in beautifying health products.