利用液相層析串聯質譜儀進行雞蛋中動物用藥Piperazine殘留之檢驗方法開發

Development of a Liquid Chromatography-tandem Mass Spectrometry Method for Analyzing Piperazine Residues in Eggs

10.6578/TJACFS.202303_61(1).0001

曾衡宇(Heng-Yu Tseng)；劉誌成(Chih-Cheng Liu)；黃志能(Chih-Neng Huang)；林汝青(Nu-Ching Lin)；高雅敏(Ya-Min Kao)；曾素香(Su-Hsiang Tseng)；王德原(Der-Yuan Wang)

Piperazine ； 動物用藥 ； 液相層析串聯質譜儀 ； BEH HILIC ； Piperazine ； Animal drug ； LC-MS/MS ； BEH HILIC

臺灣農業化學與食品科學

61卷1期（2023 / 03 / 01）

1 - 9

繁體中文;英文

本研究針對雞蛋基質，利用液相層析串聯質譜儀配合多重反應偵測（multiple reaction monitoring, MRM）模式，建立動物用藥piperazine之檢驗方法。檢體採用5%三氯醋酸溶液及乙溶液萃取，並以乙飽和之正己烷以及陽離子型固相萃取匣淨化。分離上採用BEH HILIC管柱配合含0.1%甲酸之乙溶液及含0.1%甲酸之50 mM醋酸銨溶液進行梯度沖提。分析上採用同位素內部標準品（piperazine-d_8）校正之基質匹配檢量線進行定量。本研究進行三個濃度（0.005、0.025及0.05 ppm）之添加回收試驗，結果顯示，同日中雞蛋之平均回收率介於101.2～103.5%；雞蛋基質中，同日之變異係數小於8%，異日之變異係數小於3%，符合食品藥物管理署食品化學檢驗方法之確效規範。本方法在雞蛋基質中之定量極限為0.005 ppm。本方法靈敏及穩定，可適用雞蛋基質之分析，已於109年11月6日對外公開，供外界參考並運用於後市場監測，保障消費者健康。

The aim of this study was to develop an analytical method for determining piperazine in chicken egg using liquid chromatography tandem mass spectrometry (LC-MS/MS). The pretreatment procedure was applied by extraction using 5% trichloroacetic acid and acetonitrile and following by the clean-up procedure using the Bond Elute Plex PCX cartridge and n-hexane saturated with acetonitrile. Chromatographic separation was achieved using a BEH HILIC column operated with a gradient mobile phase system consisting of acetonitrile containing 0.1% formic acid and 50 mM ammonium acetate containing 0.1% formic aicd. Quantification used the matrix-matched calibration normalized by the isotope-labeled internal standard (piperazine-d_8). The method was validated in accordance with the requirements of Taiwan validation guideline of the food chemical testing method using various concentrations (0.005, 0.025, 0.05 ppm). The intra-day average recoveries ranged from 101.2 to 103.5% for chicken egg. The precisions for all matrices were bellow 8% and 3% in intra-day and in inter-day, respectively. LOQs of piperazine in egg were 0.005 ppm. This fast, sensitive and rugged method was suitable for chicken egg. The method had been released in 2020 to the public for conducting post-market surveillance to protect consumer health.

1. European Union. Commission decision of 2002/657/EC Implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. Official Journal of the European Union, L221: 8-36 (2002).
2. Chung, S.-W.-C.,Lam, C.-H.(2015).Development of a 15-class multiresidue method for analyzing 78 hydrophilic and hydrophobic veterinary drugs in milk, egg and meat by liquid chromatography-tandem mass spectrometry.Analytical Methods,7,6764-6776.
3. Dold, C.,Holland, C. V.(2011).Ascaris and Ascariasis.Microbes and Infection,13,632-637.
4. Martin, R. J.(1985).γ-Aminobutyric acid- and piperazine-activated single channel currents from Ascaris suum body muscle.British Journal of Pharmacology,84,445-461.
5. Mc Clintic, C.,Remick, D. M.,Peterson, J. A.,Risley, D. S.(2003).Novel method for the determination of piperazine in pharmaceutical drug substances using hydrophilic interaction chromatography and evaporative light scattering detection.Journal of Liquid Chromatography & Related Technologies,26,3093-3104.
6. Park, J. A.,Zhang, D.,Kim, D. S.,Kim, S. K.,Cho, S. H.,Jeong, D.,Kim, J. S.,Shim, J. H.,El-Aty, A. M. A.,Shin, H. C.(2016).Development of a high-performance liquid chromatography with fluorescence detection method for quantification of piperazine in animal products by using precolumn derivatization.Food Chem.,196,1331-1337.
7. Staack, R. F.,Maurer, H. H.(2003).Toxicological detection of the new designer drug 1-(4-methoxyphenyl) piperazine and its metabolites in urine and differentiation from an intake of structurally related medicaments using gas chromatography-mass spectrometry.Journal of Chromatography B,798,333-342.
8. The Japan Food Chemical Research Foundation(2015).The Japan Food Chemical Research Foundation. Maximum residue limits (MRLs) list of agricultural chemicals in foods. (2015)..
9. Wang, S.,Cyronak, M.,Yang, E.(2007).Does a stable isotopically labeled internal standard always correct analyte response?.Journal of Pharmaceutical and Biomedical Analysis,43,701-707.
10. Xie, K.,Liu, Y.,Sun, L.,Pang, M.,Xie, X.,Gao, Q.,Wang, B.,Zhang, Y.,Wang, R.,Zhang, G.,Dai, G.,Wang, J.(2017).Quantification of piperazine in chicken muscle by ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry.Food Analytical Methods,10,1736-1744.
11. 衛生福利部。動物用藥殘留標準。109年9月17日衛授食字第1091303002號令修正(2020)。
12. 衛生福利部食品藥物管理署(2013)。衛生福利部食品藥物管理署。食品化學檢驗方法之確效規範。102年9月9日公開(2013)。