高效液相色谱-串联质谱法快速分析鱼肉中的类固醇激素残留

Rapid Analysis of Steroid Hormone Residues in Fish Flesh by High Performance Liquid Chromatography-tandem Mass Spectrometry

摘要:

建立了一种快速高效的液相色谱-串联质谱法分析鱼肉中类固醇激素。实验优化了色谱分离条件和三重四级杆质谱检测参数。采用Hypersil GOLD C18色谱柱（150 mm×2.1 mm，i.d. 5 μm），甲醇-水（含0.1%甲酸，V/V，%）为流动相进行梯度洗脱，在20 min内基本实现了大部分物质基线分离；在电喷雾正离子和选择离子对模式下，结合目标物保留时间和特征离子对信息，对16种激素进行定性和定量分析。结果表明，所有化合物具有良好的线性范围（0.10~3000.00 ng/mL），相关系数R大于0.9987，检测限介于0.05~5.00 ng/mL范围内。鱼肉样品在液氮中研磨成鱼粉，经过固相萃取法净化处理后，测得氢化可的松药物残留为48.00 ng/g。16种激素的回收率介于82.84%~115.10%之间，RSDs值小于9.30%。该法分离能力强、灵敏度高，适用于水产品中类固醇药物的多残留同时测定。

Abstract:

A rapid and efficient high performance liquid chromatography-tandem mass spectrometry method was established for the analysis of steroid hormones in fish flesh. The conditions for chromatographic separation and triple-quadrupole mass spectrometry detection were optimized. Baseline separation was achieved for most of the substances within 20 min using a Hypersil GOLD C18 column (150 mm×2.1 mm, i.d. 5 μm) and gradient elution with methanol-water (containing 0.1% formic acid, V/V, %). Most compounds were baseline separated in 20 min. Electrospray ionization in positive ion mode with selective ion pairing, along with the retention time and characteristic ion pairs of the target compound, was used for qualitative and quantitative analyses of 16 steroid hormones. The results showed that all compounds had a good linear range of 0.10~3000.00 ng/mL, with correlation coefficient R greater than 0.9987 and limit of detection in range of 0.05~5.00 ng/mL. Fish flesh was ground into powder in liquid nitrogen, and the latter was purified by solid phase extraction. The hydrocortisone drug residue in fish powder was 48.00 ng/g. The recoveries of 16 steroid hormones were in range of 82.84%~115.10%, with the RSD values lower than 9.30%. This method had high separation power and high sensitivity, and is suitable for simultaneous determination of steroid hormones in aquatic products.