動物產品中極性農藥及其代謝物之多重殘留分析方法開發   全文下載

Development of a Multi-Residue Analytical Method for Polar Pesticides and their Metabolites in Animal Products

本研究參考歐盟QuPPe方法（Quick Polar Pesticides Method），建立以液相層析串聯質譜法（Liquid Chromatography Tandem Mass Spectrometry, LC-MS/MS）分析蜂蜜、蛋、乳、肌肉、內臟及脂肪中8項極性農藥及其代謝物包括嘉磷塞、嘉磷塞代謝物1（Aminomethylphosphonic acid, AMPA）、嘉磷塞代謝物2（N-acetyl glyphosate）、固殺草（Glufosinate-ammonium）、固殺草代謝物1〔3-（Methylphosphinico）propionic acid, MPPA〕、固殺草代謝物2（N-acetyl-glufosinate）、益收生長素（Ethephon）及福賽得（Fosetyl-Al）之多重殘留檢驗方法。檢體前處理流程先依基質調整水分含量，添加內部標準品後即以含1%甲酸之甲醇（脂肪基質改以含1%甲酸之70%甲醇）萃取，置於-20℃冷凍90分鐘，於-10℃以5,000×g離心5-10分鐘，再依基質複雜程度搭配乙腈或C18吸附劑淨化，稀釋5-10倍後，經0.22μm PTFE濾膜過濾後以LC-MS/MS分析。本方法使用Anionic Polar Pesticide（APP）層析管柱搭配含1.2%甲酸之去離子水及含0.5%甲酸之乙腈作為移動相，可良好分離嘉磷塞等8項極性農藥及其代謝物。確效試驗以含6項內標之標準曲線進行6種代表性動物基質中8項極性農藥及其代謝物之添加回收，於添加濃度為0.05 ppm時，平均回收率為63.1-117.1%，變異係數除肝臟中嘉磷塞代謝物1（AMPA）為20.7%外，其餘皆小於16.1%；於添加濃度為0.1 ppm時，平均回收率為65.5-131.9%，變異係數皆小於14.1%。除肝臟中添加0.05 ppm之AMPA變異係數較大外，皆符合歐盟SANTE/11312/2021規範，定量極限（Limit of Quantification, LOQ）除肝臟中AMPA為0.1 ppm，其餘皆為0.05 ppm，可滿足我國動物產品中嘉磷塞殘留容許量要求，亦可接軌多數國際標準。以市售20件動物產品進行檢驗方法適用性驗證，結果均符合規定。

In this study, we developed a multi-residue analytical method for 8 polar pesticides and their metabolites, including glyphosate, aminomethylphosphonic acid (AMPA), N-acetyl glyphosate, glufosinate-ammonium, 3-(methylphosphinico)propionic acid (MPPA), N-acetyl-glufosinate, ethephon and fosetyl-aluminum, in animal products such as honey, eggs, milk, muscle, viscera and lard using LC-MS/ MS. Based on the QuPPe method published by the European Union, polar pesticides were extracted from samples by methanol containing 1% formic acid except lard matrix of which 70% methanol containing 1% formic acid was used following water adjustment and isotope-labelled internal standards (IL-IS) addition. After freezing out for 90 minutes at -20℃and centrifugation at 5000×g at -10℃for 5-10 minutes, the extracts were cleaned up with acetonitrile or C18 absorbent depending on the matrix, then analyzed by LCMS/ MS after the supernatant was diluted 5-10 folds and filtrated by 0.22μm PTFE. In this study, eight polar pesticides and their metabolites were well separated using an Anionic Polar Pesticide (APP) column with deionized water containing 1.2% formic acid and acetonitrile containing 0.5% formic acid as the mobile phase. Using solvent-based calibration curves with 6 isotope-labelled internal standards (IL-IS), the validation results showed the recoveries of 8 polar pesticides and their metabolites in 6 animal matrices were 63.1-117.1% at the spiked level of 0.05 ppm, and the coefficients of variation were all less than 16.1% except that of AMPA was 20.7% in liver. At the spiked level of 0.10 ppm, the recoveries were 65.5-131.9%, and CVs were all less than 14.1%. The above validation data of 8 polar pesticides and their metabolites in 6 animal matrices was in accordance with the SANTE/11312/2021 at the spiked level of 0.05 ppm except for AMPA in liver. The limits of quantification (LOQs) were all 0.05 ppm except for AMPA in viscera which was 0.1 ppm. This method meets the requirement of residue limits in Taiwan and also in line with most international standards. Using 20 commercial animal products to verify the applicability of the method, the results showed that all samples were in compliance with the regulation.