超高液相層析串聯質譜法檢測豬組織中的16種乙型受體素

Determination of 16 β-agonists in Pork Tissue by Ultra High Liquid Chromatography/Tandem Mass Spectrometry

黃志能(Chih-Neng Huang) ； 趙偉博(Wei-Po Jau) ； 於柏伸(Bo-Shen Wu) ； 曾素香(Su-Hsiang Tseng) ； 高雅敏(Ya-Min Kao) ； 闕麗卿(Lih-Ching Chiueh) ； 施養志(Daniel Yang-Chih Shih)

《臺灣農業化學與食品科學》 52卷1期 (2014/02) Pp. 31-47

https://doi.org/10.6578/TJACFS.2014.005

摘要

本研究建立以超高效液相層析串聯質譜法如（utra high liquid chromatography/tandem mass spectrometer, LC/MS/MS）利用正離子電灑法以多重反應偵測（multiple reaction monitoring, MRM）模式同時分析豬肉、豬肝及豬腎中16項乙型受體素之檢驗方法，檢體經由0.2M醋酸鈉緩衝液萃取後，配合酵素水解、酸沈澱蛋白質及陽離子固相萃取匣的淨化處理，其檢液以氮氣吹乾，以5mM醋酸銨：甲醇（9:1, ν/v）溶液回溶並經由高速離心後，取上清液，使用ZORBOX RRHD Eclipse Plus C18 Coiumn, 18μm, 3.0x100mm層析管柱，以0.1%甲酸溶液與當0.1%甲酸之乙睛溶液為移動相進行梯度流洗，再以串聯質譜儀分析。並運用內標校正後之基質匹配檢量線（matrix-matched calibration curve）進行定量。結果顯示，豬肉方面，在添加2、5、10 ng/g濃度下，16項藥物之平均回收率為73.9-130.7%，變異係數為4.3-27.2%；內臟方面，在添加10、20、40 ng/g濃度下，16項藥物之平均回收率77.8-135.1%，變異係數為2.7-23.4%。本方法之定量極限為2-10 ng/g。

關鍵字

畜禽產品 ； 超高效液相層析串聯質譜儀 ； 多重殘留分析 ； 乙型受體素

並列摘要

A method for the analysis of 16 β-agonists in pork, swine liver and swine kidney by ultra high liquid chromatography/tandem mass spectrometer (LC/MS/MS) with electrospray ionization was developed. Data acquisition under MS/MS for each β-agonists was achieved by applying multiple reaction monitoring (MRM). The sample was extracted with 0.2 M sodium acetate buffer, followed by hydrolyzsis with enzymes and protein precipitation with acid. After clarified through cation exchange solid-phase extratction, the analyte was dried with nitrogen gas. After resuspensing in 5 mM ammonium acetate:methanol (9 : 1, v/v), the samples were centrifugated with high speed to obtain supernant. The LC separation was conducted on an ZORBOX RRHD Eclipse Plus C18 Column (1.8 μm, 3.0 × 100 mm) with gradient elution of 0.1% formic acid in water and acetonitrile. The recoveries of 16 β-agonists (2, 5 and 10 ng/g) spiked in pork ranged from 73.9 to 130.7% and the coefficients of variation of all β-agonists were between 4.3 to 27.2%. In swine liver and swine kidney, the recoveries of 16 β-agonists (10, 20 and 40 ng/g) ranged from 77.8 to 135.1% and the coefficients of variation of all β-agonists were between 2.7 to 23.4%. The quantitation limit of the above method is 2 to 10 ng/g.

並列關鍵字

animal products ； ultra high liquid chromatography/tandem mass spectrometer ； multiresidues analysis ； β-agonists