液相色谱-四极杆/离子阱质谱同时确证和测定肌肉中16种同化甾体激素残留

采用液相色谱-四极杆/离子阱质谱(LC-Q/Trap-MS)建立了肌肉中16种同化甾体激素类物质(ASs)残留的同时确证及测定方法。肌肉中的ASs采用乙腈超声辅助提取,正己烷脱脂,氨基固相萃取柱净化,CAPCELL PAK C18 MGIII柱(150 mm×2.0 mm, 5.0 μm)分离,0.1%(v/v)甲酸-乙腈溶液和0.1%(v/v)甲酸-5 mmol/L甲酸铵水溶液为流动相梯度洗脱;预设定多反应监测(sMRM)-信息依赖性采集(IDA)-增强子离子扫描(EPI)模式检测,在线EPI谱库确证,内标法定量。结果表明,16种ASs在线性范围内线性关系良好(r≥0.999);定量限(LOQ, S/N≥10)为0.029～0.36 μg/kg; 3个添加水平(0.5、2.0和20 μg/kg)下的回收率为89.9%～118%;相对标准偏差(RSD)为6.3%～16.2%。该方法准确灵敏,一次性完成16种ASs的确证和测定,可有效用于肌肉组织中ASs残留的监测分析。

Simultaneous identification and detection of 16 anabolic steroid hormones in muscle using liquid chromatography coupled to quadrupole/linear ion trap mass spectrometry

A comprehensive method for simultaneous identification and detection of 16 anabolic steroid hormones (ASs, including andorgens, gestagens and their esters)in muscle samples was developed with liquid chromatography coupled to quadrupole/linear ion trap mass spectrometry (LC-Q/Trap-MS). The ASs in muscle samples were extracted with acetonitrile under ultrasonic assistance. The extract was defatted by n-hexane with liquid-liquid partitioning and followed by clean-up with NH2 solid phase extraction (SPE) cartridge. The separation of analytes was carried out on a CAPCELL PAK C18 MGIII column (150 mm×2.0 mm, 5.0 μm) using mobile phases of 0.1% (v/v) formic acid in acetonitrile and 0.1% (v/v) formic acid-5 mmol/L ammonium formate aqueous solution with gradient elution. A scheduled multiple reaction monitoring (sMRM) in positive mode as survey scan and an enhanced product ion (EPI) scan as dependent scan in an information-dependent acquisition (IDA) experiment was adopted in mass spectrometry acquisition. On-line lab-built MS/MS library and internal standards were employed for the identification and quantification. As a result, the 16 ASs showed good linearity with all correlation coefficients (r) no less than 0.9990 within the linear ranges. The limits of quantification (LOQs, S/N≥10) for the 16 ASs were in the range of 0.029~0.36 μg/kg. At the three spiked levels (0.5, 2.0 and 20 μg/kg), the overall recoveries ranged from 89.9% to 118% with the relative standard deviations (RSDs) no more than 16.2% under within-laboratory reproducibility conditions. The proposed method can be used to identify and detect the 16 ASs in a single run, which makes it effective in residue surveillance of anabolic hormones in muscle samples.