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-d8)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-d8). 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.