食品科学 ›› 2009, Vol. 30 ›› Issue (14): 204-206.doi: 10.7506/spkx1002-6630-200914041

• 分析检测 • 上一篇    下一篇

同时适用ELISA和GC/MS快速测定猪肉中盐酸克伦特罗前处理方法研究

吴银良,杨 挺,朱 勇,赵 剑,皇甫伟国   

  1. 农业部农产品质量安全监督检验测试中心(宁波)
  • 收稿日期:2008-10-17 修回日期:2009-01-16 出版日期:2009-07-15 发布日期:2010-12-29
  • 通讯作者: 吴银良 E-mail:wupaddyfield@tom.com
  • 基金资助:

    农业部2008 年农业行业标准资助专项(2008259)

A Pre-treatment Method Suitable for ELISA and GC-MS Determination of Clenbuterol in Pork

WU Yin-liang,YANG Ting,ZHU Yong,ZHAO Jian,HUANGFU Wei-guo   

  1. Supervision, Inspection and Testing Center of Agricultural Products Quality and Security, Ministry of Agriculture, Ningbo, Ningbo
    315040, China
  • Received:2008-10-17 Revised:2009-01-16 Online:2009-07-15 Published:2010-12-29
  • Contact: WU Yin-liang, E-mail:wupaddyfield@tom.com

摘要:

建立同时适用于ELISA 和GC-MS 方法快速测定猪肉中盐酸克伦特罗残留量的前处理方法。猪肉样品在碱化的条件下用乙酸乙酯提取,再用稀盐酸萃取去除脂肪,调pH 值后再经乙酸乙酯反萃取,乙酸乙酯反萃取液均分成两份用氮气吹干,一份加水用于ELISA 检测,另一份经双三甲基硅基三氟乙酰氨(BSTFA)衍生,采用选择离子模式(m/z86、212、262、277)进行测定,外标法定量,GC-MS 检出限为0.30μg/kg。在加标浓度为1.0、2.0、5.0μg/kg 时,加标回收率为72.1%~105.6% (ELISA)和79.1%~95.7% (GC-MS),批间相对标准偏差(RSD)为12.5%~16.8% (ELISA)和4.6%~6.9% (GC-MS)。衍生物的峰面积与样品浓度在0. 005~1.000mg/L 范围内呈良好的线性关系,线性回归系数大于0.999。

关键词: 猪肉, 酶联免疫分析, 气相色谱- 质谱法, 盐酸克伦特罗, 残留

Abstract:

A pre-treatment method was developed for determining residual clenbuterol in pork by ELISA and GC-MS. Samples were extracted with ethyl acetate under basic condition, and the extracts were further extracted using diluted hydrochloric acid to remove fat. The hydrochloric acid extracts were re-extracted using ethyl acetate following pH adjustment, and the final extract obtained was divided into two equal parts and each of them was dried by nitrogen blow. Water was added to one part prior to ELISA determination. After derivatization with bis(trimethylsilyl)trifluoroacetamide (BSTFA) another part was detected by GC-MS in selected ion monitoring mode (selected ions: 86, 212, 262, 277) and quantitatively analyzed by external standard method. The GC-MS detection limit was 0.30 μg/kg. The recoveries for clenbuterol determined by ELISA in pork spiked at the levels of 1.0 -5.0 μg/kg were 72.1%-105.6%, and by GC-MS 79.1%-95.7%, and the intra-day relative standard deviations for ELISA and GC-MS determination were 12.5%- 16.8% and 4.6%- 6.9%, respectively. There was a good linear correlation (the calibration coefficient was above 0. 999) between peak area and concentration of clenbuterol derivative in the range of 0.005 to 1.000 mg/L.

Key words: pork, ELISA, gas chromatography-mass spectrometry, clenbuterol, residue

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