色谱

色谱 ›› 2012, Vol. 30 ›› Issue (10): 991-1001.DOI: 10.3724/SP.J.1123.2012.08017

• 专栏 • 上一篇    下一篇

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

张鸿伟1*, 蔡雪1, 林黎明2, 陈亮珍3, 梁成珠1,鲍 蕾1, 汤志旭1, 牛增元2, 王凤美1   

  1. 1. 山东出入境检验检疫局检验检疫技术中心, 山东 青岛 266002; 2. 山东省检验检疫科学技术研究院, 山东 青岛 266002; 3. 青岛蔚蓝生物集团, 山东 青岛 266061
  • 收稿日期:2012-08-15 修回日期:2012-08-31 出版日期:2012-10-28 发布日期:2012-10-17
  • 通讯作者: 张鸿伟,博士在读,高级工程师,研究方向为食品中兽药残留分析. E-mail: light04@126.com.
  • 基金资助:

    国家质检总局科技项目(2007IK144)、山东省科技发展计划项目(2008GG10009020)和山东出入境检验检疫局科研攻关项目(SK200820).

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

ZHANG Hongwei1*, CAI Xue1, LIN Liming2, CHEN Liangzhen3, LIANG Chengzhu1, BAO Lei1, TANG Zhixu1, NIU Zengyuan2, WANG Fengmei1   

  1. 1. Technical Center of Entry-Exit Inspection and Qurantine, Shandong Entry-Exit Inspection and Quarantine Bureau, Qingdao 266002, China; 2. Science and Technology Institute of Shandong Quarantine and Inspection, Qingdao 266002, China; 3. Qingdao Vland Biotech Group, Qingdao 266061, China
  • Received:2012-08-15 Revised:2012-08-31 Online:2012-10-28 Published:2012-10-17

PDF

424

被引次数

21

摘要： 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。

关键词: 残留, 肌肉, 四极杆/离子阱质谱, 同化甾体激素, 液相色谱

Abstract: 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.

Key words: anabolic steroid hormones (ASs), muscle, quadruple/linear ion trap mass spectrometry (Q/Trap-MS), residues, liquid chromatography (LC)