同位素稀释-超高效液相色谱-串联质谱法测定鸡蛋中3种类固醇激素

Quantification of 3 kinds of steroid hormones in eggs by isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry

中文关键词:  同位素稀释法  超高效液相色谱-串联质谱法  禽产品  类固醇激素  基质效应

英文关键词:isotope dilution method  ultra performance liquid chromatography-tandem mass spectrometry  poultry products  steroid hormone  matrix effect

中文摘要:

      目的 建立一种同位素稀释-超高效液相色谱-串联质谱法(isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry, ID-UPLC-MS/MS)检测禽产品中典型类固醇激素(雌酮、孕酮和睾酮)的分析方法。方法 样品经乙腈提取, 甲醇洗脱后, 氮吹至干燥后用1 mL甲醇复溶, 上机检测。用0.1%甲酸水溶液(A)和甲醇(B)作流动相进行梯度洗脱, 在电喷雾正离子模式和多反应监测模式下采集离子对, 检测目标物。结果 建立的方法在5 min内完成了目标化合物(雌酮、孕酮和睾酮)的定量分析。3种类固醇激素在0.5~100.0 μg/kg范围内线性关系良好, r2均大于0.996。雌酮、孕酮和睾酮的方法检出限分别为0.052、0.023和0.048 μg/kg, 定量限分别为0.173、0.077和0.160 μg/kg。在1、5和50 μg/kg添加水平下, 3种目标化合物的平均回收率在96.62%~103.11%, 精密度在1.36%~3.87%之间。结论 该方法简便快速、回收率高、稳定性好, 可有效测定禽产品中雌酮、孕酮和睾酮含量。

英文摘要:

      Objective To establish a method for the quantification of 3 kinds of typical steroid hormones (estrone, progesterone and testosterone) in poultry products by isotope dilution-ultra performance liquid chromatography-tandem mass spectrometry (ID-UPLC-MS/MS). Methods The sample was extracted by acetonitrile, eluted by methanol, blown by nitrogen to dry, redissolved with 1 mL methanol, and tested on the machine. Gradient elution was performed with 0.1% formic acid aqueous solution (A) and methanol (B) as mobile phases, ion pairs were collected by electrospray positive ionization mode, and quantitative and qualitative ions were determined by multiple reaction monitoring mode. Results The established method completed the quantitative analysis of target compounds (estrone, progesterone and testosterone) within 5 min. The 3 kinds of steroid hormones had good linear relationships in the range of 0.5?100.0 μg/kg with the correlation coefficients r2 more than 0.996. The limits of detection for estrone, progesterone and testosterone were 0.052, 0.023 and 0.048 μg/kg, respectively, and the limits of quantification were 0.173, 0.077 and 0.160 μg/kg, respectively. At the addition levels of 1, 10 and 50 μg/kg, the average recoveries of the 3 kinds of steroid hormones were 96.62%?103.11%, and the precision was between 1.36% and 3.87%. Conclusion The method is simple, rapid with high recovery rates and good stability, which can effectively determine the content of estrone, progesterone and testosterone in poultry products.