超高效液相色谱-串联质谱法测定猪肝中的17-β-雌二醇

Determination of 17-β-estradiol in pork liver by ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  超高效液相色谱-串联质谱法  17-β-雌二醇  猪肝

英文关键词:ultra performance liquid chromatography-tandem mass spectrometry  17-β-estradiol  pork liver

中文摘要:

      目的 建立超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)测定猪肝中17-β-雌二醇(17-β-estradiol, E2)残留量的检测方法。方法 猪肝样品经β-葡萄糖醛苷酶/芳基硫酸酯酶酶解后用25.0 mL甲醇提取, 加入2.0 g氯化锌除脂, 用25.0 mL水溶液稀释后上样液过ENVI-Carb柱, 用5.0 mL甲醇淋洗, 二氯甲烷:甲醇(7:3, V:V)洗脱后经氨基柱进一步净化, 以甲醇-0.50 mmol/L氟化铵作为流动相, 经ACQUITY UPLC BEH C18 (50 mm×2.1 mm, 1.7 μm)色谱柱分离后, 在电喷雾负离子多反应监测扫描(multiple reaction monitoring, MRM)模式下进行检测。结果 E2的线性范围为0.01~100.00 μg/L, 相关系数r2>0.999, 检出限为7 ng/kg, 定量限为20 ng/kg, 添加水平为0.02、0.05和0.10 μg/kg时的平均回收率为101.1%~106.7%, 日内精密度为5.1%~7.3% (n=6), 日间精密度为10.0%。应用建立的方法对5份猪肝样品进行测定, 所有样品均检出E2。结论 该方法具有灵敏度高、操作简单快速的特点, 适用于猪肝中E2残留量的检测。

英文摘要:

      Objective To establish a method for the determination of 17-β-estradiol (E2) residues in pork liver by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Methods The pork liver sample was hydrolyzed by β-glucosidase/aryl sulfatase and extracted with 25.0 mL methanol, and 2.0 g zinc chloride was added to remove the grease, after diluted with 25.0 mL aqueous solution, the sample solution was passed through an ENVI-Carb column and eluted with 5.0 mL methanol, after eluted with dichloromethane: Methanol (7:3, V:V), the sample was further purified by an amino column with methanol-0.50 mmol/L ammonium fluoride as the mobile phase, and the assay was conducted in an electrospray negative ion multiple reaction monitoring (MRM) mode after separation on an ACQUITY UPLC BEH C18 (50 mm×2.1 mm, 1.7 μm) column. Results The linear range of E2 was from 0.01 to 100.00 μg/L, with the correlation coefficient r2>0.999, the limit of detection was 7 ng/kg, and the limit of quantitation was 20 ng/kg, the average recoveries at the addition levels of 0.02, 0.05, and 0.10 μg/kg were 101.1%?106.7%, and the intra-day precision was 5.1%?7.3% (n=6), and the intra-day precision was 10.0%. Five pork liver samples were determined by the established method, and E2 was detected in all samples. Conclusion This method is sensitive, simple and rapid, and suitable for the determination of E2 residues in pork liver.