多重机制杂质吸附萃取净化-快速液相色谱-串联质谱法测定鱼组织中11种同化激素

Determination of 11 anabolic hormones in fish tissue by multi-function impurity adsorption solid-phase extraction-ultrafast liquid chromatography-tandem mass spectrometry

建立了准确、灵敏的鱼组织中11种同化激素(勃地酮、雄烯二酮、诺龙、美雄酮、甲睾酮、睾酮、醋酸睾酮、群勃龙、丙酸睾酮、康力龙、氟甲睾酮)的多重机制杂质吸附萃取净化-快速液相色谱-串联质谱的分析方法。鱼组织均质样品经甲醇提取后,在上清液中加入一定量的C18固体吸附剂、中性氧化铝吸附剂和氨基功能化纳米吸附剂实现快速净化。采用Shim-Pack XR-ODSⅡ色谱柱(100 mm×2.0 mm,2.2μm)分离,以乙腈(含0.1%甲酸)和水(含0.1%甲酸)为流动相进行梯度洗脱,电喷雾正离子多反应监测(MRM)模式下检测,外标法定量。结果表明,11种目标化合物在线性范围内具有良好的线性关系,相关系数大于0.999,其在鱼组织中的检出限(S/N>3)为0.03～0.4μg/kg,定量限(S/N>10)为0.1～1.5μg/kg,平均回收率为80.9%～98.1%,相对标准偏差(RSD)为5.2%～11.5%。该方法简便、快速、准确,可用于鱼组织中同化激素的定性、定量监测。

A method was developed for the determination of 11 anabolic hornlones （bolde- none, androstenedione, nandrolone, methandrostenolone, methyltestosterone, testosterone, testosterone acetate, trenbolone, testosterone propionate, stanozolol, fluoxymesterone） in fish by multi-function impurity adsorption solid-phase extraction-ultrafast liquid chromatography- tandem mass spectrometry. After the sample was extracted by methanol, the extract was cleaned-up quickly by C18 adsorbent, neutral alumina adsorbent and amino-functionalized nano- adsorbent. The separation was performed on a Shim-Pack XR-ODS Ⅱ column （ 100 mm ×2.0 nun, 2.2 pure） using the mobile phases of 0. 1% （v/v） formic acid in acetonitrile and 0. 1% （v/v） formic acid solution in a gradient elution mode. The identification and quantification were achieved by using electrospray ionization in positive ion mode （ ESI^＋ ） in multiple reaction monitoring （MRM） mode. The matrix-matched external standard calibration curves were used for quantitative determination. The results showed that the calibration curves were in good line- arity for the eleven analytes with the correlation coefficients （r） more than 0. 999. The limits of detection （ LODs, S/N 〉 3） for the I I anabolic hormones were from 0.03 μg/kg to 0.4 μg/kg and the limits of quantification （LOQs, S/N 〉 10） were from 0. 1 μg/kg to 1. 5 vug/kg. The average recoveries ranged from 80.9% to 98. 1% with the relative standard deviations between5.2% and 1 1.5%. The method is simple, rapid, sensitive, accurate and suitable for the quanti- tative determination and confirmation of the 1 t anabolic hormones in fish.