穀類中多重黴菌毒素檢驗方法之建立

Simultaneous Determination of Multi-Mycotoxin in Cereals Using LC-MS/MS

陳映君(Ying-Chun Chen) ； 廖家鼎(Chia-Ding Liao) ； 王璽權(Si-Chuan Wang) ； 林旭陽(Hsu-Yang Lin) ； 闕麗卿(Lih-Ching Chiueh) ； 羅吉方(Chi-Fang Lo)

《食品藥物研究年報》 2期 (2011/12) Pp. 164-171

https://doi.org/10.6945/ARFDR.201112.0166

摘要

黴菌毒素會造成人體肝臟與腎臟毒性，影響民眾健康甚鉅，衛生署也已訂定食品中多種黴菌毒素之限量標準。本研究針對數種常見之黴菌毒素，包括黃麴毒素B1（Aflatoxin B1，AFB1）、黃麴毒素B2（Aflatoxin B2，AFB2）、黃麴毒素G1（Aflatoxin G1，AFG1）、黃麴毒素G2（Aflatoxin G2，AFG2）、伏馬毒素B1（Fumonisin B1，FB1）、伏馬毒素B2（Fumonisin B2，FB2）、脫氧雪腐鐮刀菌烯醇（Deoxynivalenol，DON）、玉米赤黴毒素（Zearalenone，ZON）、赭麴毒素A（Ochratoxin A）、T-2毒素與HT-2毒素等，開發以液相層析串聯式質譜（LC/MS/MS）同步分析多種黴菌毒素之方法，可大幅改進傳統HPLC方法之分析效率。在樣品前處理方面，以磷酸緩衝溶液及70%甲醇做二次萃取，可對較親水性的毒素得到較高之回收率，DON效果尤為顯著。另以免疫親和管柱Myco6in1（上標 TM）淨化，減少基質干擾。利用ACQUITY BEH C18層析管柱，移動相以含有0.1%甲酸之去離子水與甲醇做梯度分離可得到最適之峰型。所有毒素之偵測極限皆可至1ppb以下。以白米、花生、小麥和玉米為基質做各毒素之添加回收率試驗結果，各毒素之平均回收率良好，大都介於65－120%之間，變異係數皆小於20%。本方法之檢出限量分別為0.5ppb（AFB1）、0.5ppb（AFB2）、0.5ppb（AFG1）、0.5ppb（AFG2）、5ppb（ZON）、1ppb（T-2）、1ppb（HT-2）、0.5ppb（OTA）、20ppb（FB1）、20ppb（FB2）、5ppb（DON）。本方法可在9分鐘內快速同步分析11種黴菌毒素，可實際應用於食品中黴菌毒素之監測，以保障民眾健康。

關鍵字

黴菌毒素 ； 液相層析串聯式質譜 ； 同步分析 ； 免疫親和管柱

並列摘要

Mycotoxins are secondary metabolites produced by fungi that are capable of causing disease and even death in humans and other animals. A multi-mycotoxin analytical method was developed for the simultaneous determination of aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2), fumonisin B1 (FB1), fumonisin B2 (FB2), deoxynivalenol (DON), zearalenone (ZON), ochratoxin A (OTA), T-2 and HT-2 toxin in cereals. Cereals were extracted with PBS and 70% methanol, cleaned up using immunoaffinity columns and analyzed by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) with an electrospray (ESI) ionization interface. Using the positive ion mode, the parent and product ions of each mycotoxin were selected. The LC separation was performed using ACQUITY UPLC BEH C18 column with the mobile phase consisting of gradient program of water and methanol with 0.1% formic acid. Besides FB1 and FB2, the limit of quantification (LOQ) for most mycotoxins can achieve below 5 ppb. Good recoveries were found among various mycotoxins, ranging from 65 to 120%. The coefficient of variance (CV) was below 20%, indicating this method is reliable. This rapid and easy method was successfully developed to detect eleven mycotoxins in a single 9-min run. The method could be applied for routine cereal analysis in order to safeguard people's health.

並列關鍵字

mycotoxins ； LC/MS/MS ； simultaneous determination ； immunoaffinity column