超高效液相色谱-串联质谱法同时测定奶粉中8种雌激素残留

目的： 建立超高效液相色谱-串联质谱法（UPLC-MS/MS法）同时测定奶粉中8种雌激素残留。方法： 0.5 g奶粉样品用2.0 mL温水溶解，经8.0 mL 0.1%甲酸-乙腈提取，通过离心去除沉淀后，将上清液过Waters oasis Prime-HLB（6 CC，200 mg）柱净化，收集洗脱液于40℃氮吹至近干，复溶后采用Waters CORTECS T₃（2.1 mm×50 mm，2.7 μm）色谱柱，以0.1%氨水溶液和乙腈为流动相进行梯度洗脱。在电喷雾离子源负离子（ESI^-）模式下电离，以多反应监测模式（MRM）进行质谱检测。结果： 8种激素残留在定量下限到20倍定量下限范围内有良好的线性关系，相关系数（R²）均大于0.99，检测下限（LOD）为0.33~2.07 μg·kg^-1，定量下限（LOQ）为0.98~6.21 μg·kg^-1，3种加标水平下平均回收率在95.2%~118.1%之间，RSD（n=6）均小于6.7%。采用已建立的方法，对10批市售奶粉样品进行检测，结果样品中均含有内源性雌激素，检出β-雌二醇、α-雌二醇、雌酮。结论： 实验所建立方法简便，灵敏，实用性强，可同时测定牛奶中的8种雌激素残留，适用于日常监测工作。

Objective: To establish a method for the simultaneous determination of 8 estrogen residues in milk powder by UPLC-MS/MS. Methods: 0.5 g milk powder sample was dissolved in 2.0 mL warm water,the target compounds was extracted by 8.0 mL 0.1% formic acid-acetonitrile, the precipitate was removed by centrifugation,and the clear supernatant extract was purified by Waters oasis Prime-HLB(6 CC,200 mg). The separation and analysis were performed on a Waters CORTECS T₃(2.1 mm×50 mm,2.7 μm). The mobile phase consisted of 0.1% ammonia aqueous solution and acetonitrile with gradient elution. Ionization was conducted under the negative ion (ESI^-) mode of the electrospray ion source,and mass spectrometry was detected by multiple reaction monitoring mode (MRM). Results: Good linear relationships were obtained between the quantitative limits and 20-fold quantitative limits of 8 estrogens. The correlation coefficients (R²) were greater than 0.99,the limit of detection (LOD) was 0.33-2.07 μg·kg^-1,the limit of quantitative(LOQ) was 0.98-6.21 μg·kg^-1,the average recoveries were 95.2%-118.1% at the three levels,and the relative standard deviations (n=6) below 6.7%. The established method was used to test 10 batches of marketable milk power. The results showed that all the samples contained 17-β-estadiol,17-α-estradiol and estrone. Conclusion: The established method is simple,sensitive and practical,and suitable for the simultaneous determination of 8 estrogen residues in milk power.

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