Simultaneous Quantification of Aflatoxins, Ochratoxin a and Zearalenone in Cereals by LC-MS/MS

黃麴毒素、赭麴毒素A及玉米赤黴毒素

廖家鼎(Chia-Ding Liao) ； 林旭陽(Hsu-Yang Lin) ； 闕麗卿(Lih-Ching Chiueh) ； 施養志(Daniel Yang-Chih Shih)

《藥物食品分析》 19卷3期 (2011/09) Pp. 259-268

摘要

食品及飼料中黴菌毒素之污染具毒性及致癌性，嚴重威脅人類及動物健康。在多種黴菌毒素中，以黃麴毒素(AFs)、赭麴毒素A(OTA)及玉米赤黴毒素(ZON)最重要，因其在世界上具有較多污染案例。本研究開發簡便、快速的多重分析方法，同步定量AFB1、AFB2、AFG1、AFG2、OTA及ZON。以免疫親和性管柱淨化，再以液相層析串聯質譜儀分析。穀類樣品經80%甲醇萃取、免疫親和性管柱AOZ(上標 TM)淨化後，毒素籍由甲醇沖提出，以電灑游離質譜中選擇反應模式定量之。移動相爲甲醇－水，流速爲0.3mL/min。穀類基質添加AFs、OTA及ZON標準品之平均回收率分別爲65-95%，66-83%及69-86%。同日內與異日間之準確度及精確度皆理想。AFB1、AFB2、AFG1、AFG2、OTA及ZON之方法偵測極限分別爲0.1、0.2、0.1、0.3、0.03及2ppb。應用上述方法檢測市售榖類制品35件（小麥13件、燕麥7件、米6件及玉米9件），結果有4件檢出OTA，含量介於0.05-0.07ppb；2件檢出ZON，含量介於2.73-4.73ppb。本研究成功開發快速簡便且高效率之檢驗方法。可於20分鐘內同步分析6種常見黴菌毒素，此法可應用在例行性穀類檢驗，大幅縮短分析時間。

關鍵字

黃麴毒素 ； 赭麴毒素A ； 玉米赤黴毒素 ； 穀類 ； 液相層析串聯質譜 ； 同步分析

並列摘要

Thu occurrence of mycotoxins in foods and feeds has long been recognized as a potential hazard for human and animal health due to its severe toxic and carcinogenic properties. Among various mycotoxins, aflatoxins (AFs), ochratoxin A (OTA) and zearalenone (ZON) are most important because of many contamination cases in the world. In this study, a simple and rapid multiresidue method to quantify AFB1, AFB2, AFG1, AFG2, OTA and ZON was developed. The simultaneous determination of these 6 mycotoxins was performed using immunoaffinity column for clean-sp and liquid chromatography/electrospray tandem mass spectrometry (LC-MS/MS) for quantification. Cereal samples were extracted with 80% methanol followed by a Vicam AOZ(superscript TM) immunoaffinity column clean-up. Mycotoxins were eluted from the column with methanol and quantified using Selected Reaction Monitoring (SRM) mode in LC-VIS/MS with an electrospray ionization (ESI) interface. A mobile phase of methanol-water was used. Average recoveries of AFs, OTA and ZON spiked in cereals ranged 65-95%, 66-83% and 69-86%, respectively. Good accuracy and precision results were also obtained in intra-day and inter-day analysis. The limit of quantitation (LOQ) of AFB1, AFB2, AFG1, AFG2, OTA and ZON were 0.1, 0.2, 0.1, 0.3, 0.03 and 2 ppb, respectively. Thirty-five commercial cereal products including thirteen wheat samples, seven oat samples, six rice samples and nine corn samples were analyzed. The results showed that OTA was detected in four samples ranging from 0.05 to 0.07 ppb, and ZON was detected in two samples ranging front 2.73 to 4.73 ppb. This rapid, easy and highly efficient method was successfully developed. Six common mycotoxins could be analyzed in a single 20-min run. The method could be applied to routine cereal analysis, thus dramatically shortening the analysis time.

並列關鍵字

aflatoxin ； ochratoxin A ； zearalenone ； cereals ； LC-MS/MS