禽畜產品中乙型受體素多重殘留檢驗方法之精進

Improvement of a Multi-residue Analysis Method for the Determination of β-agonists in Poultry and Livestock Tissues

禽畜產品中乙型受體素殘留為民眾關心之重要議題，本研究針對肌肉（雞肉及豬肉）與內臟（豬肝及豬腎）中21種乙型受體素開發快速且準確的檢驗方法。樣品先以0.2M醋酸鈉萃取，並加入β-葡萄糖醛酸苷酶水解，經固相萃取匣（solid phase extraction, SPE）萃取及淨化後，以超高效液相層析串聯質譜儀搭配正離子電灑法（UPLCESI^+-MS／MS）進行分析，並以18項同位素內標進行校正。使用C18管柱進行梯度層析，以含0.1％甲酸的水和甲醇作為動相。結果顯示，肌肉及內臟定量極限分別為1ppb及5ppb，回收率分別介於71.9－120.6％（變異係數2.2－18.5％）及77.7－116.8％（變異係數0.6－9.0％）。本方法僅用少量樣品即可於一次分析中檢驗21種乙型受體素，且在肌肉和內臟基質中皆有良好確效結果，為一適用範圍廣、快速且有效率的乙型受體素檢驗方法，將來可應用於邊境及後市場之檢驗，保護國人食用動物性食品之安全。 β-agonists residues in poultry and livestock products has been an issue of public concerns. In order to improve the quality of the official testing method, this study developed a rapid and accurate method for the simultaneous analysis of 21 β-agonists in chicken, pork, swine liver and swine kidney. Samples were extracted by 0.2 M sodium acetate with β-glucuronidase for hydrolysis, followed by clean-up using solid phase extraction (SPE). Detection was carried out by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with 18 internal standards. Chromatographic separation was performed on a reversed phase C18 column, using a binary gradient comprised of 0.1% formic acid in water (mobile phase A) and 0.1% formic acid in methanol (mobile phase B). The results showed that the limits of quantification in muscle and viscera were 1 and 5 ppb, respectively. The recoveries of the 21 β-agonists spiked in muscle ranged from 71.9 to 120.6% and the coefficients of variation were between 2.2 and 18.5%. In viscera, the recoveries of the 21 β-agonists ranged from 77.7 to 116.8% and the coefficients of variation were between 0.6 and 9.0%. The developed method used minimal sample preparation and the 21 β-agonists can be analyzed in a single run. With the satisfactory validation results, this method can be applied to border inspection and post-market surveillance.