香辛料中黃麴毒素之檢驗方法開發及調查

Method Development and Survey of Aflatoxins in Spices

卓憲駿(Hsien-Chun Cho)；廖家鼎(Chia-Ding Liao)；曾素香(Su-Hsiang Tseng)；周秀冠(Hsiu-Kuan Chou)；陳惠芳 (Hwei-Fang Cheng)

黃麴毒素 ； 香辛料 ； aflatoxins ； spices

食品藥物研究年報

6期（2015 / 12 / 01）

109 - 117

繁體中文

黃麴毒素(Aflatoxin B_1、B_2、G_1、G_2)是黴菌產生之二級代謝產物，主要由Aspergillus flavus及A. parasiticus所產生，具有肝毒性及致癌性，經常會污染花生、玉米、米、麥類、豆類及香辛料等食品，對民眾健康造成危害。日常食品中常見胡椒粉、咖哩粉、辣椒粉等所謂「香辛料食品」，在加工過程中，或於原料儲存、生產環境及保存條件不佳等因素，皆會造成黃麴毒素污染的風險。本研究以香辛料基質為探討對象，用80%甲醇溶液為萃取溶液，10% Tween-20為稀釋液，檢體經均質萃取、過濾、免疫親合管住淨化後，續以高效液相層析儀(HPLC)分析。以45%甲醇為移動相做等度分離，流速為1.0 mL/min，層析管柱為Cosmosil 5C18-AR (5 μm，4.6× 250 mm)，螢光檢出器的激發波長為360 nm，放射波長為440 nm，並搭配光化學反應器做管柱後衍生化，分析時間為20分鐘。黃麴毒素B_1、B_2、G_1及G_2之標準曲線R^2皆大於0.995，顯示線性良好。以胡椒粉、辣椒粉及咖哩粉為基質進行回收率添加試驗，添加2、4及6 ppb各毒素標準品，各毒素之平均回收率介於61.85-101.34%之間(n=3)，變異係數(CV%)皆小於12.05%；方法定量極限(LOQ)，黃麴毒素B_1及G_1均為1ppb，黃麴毒素B_2及G_2均為0.5 ppb。顯示其良好的準確度、精密度及靈敏度，可應用於實際產品的檢驗。以此方法抽驗市售香辛料檢體共33件，包含胡椒粉10件、咖哩粉6件、辣椒粉11件及五香粉6件，結果有2件咖哩粉及4件辣椒粉檢出黃麴毒素B1，含量為0.79-2.74 ppb，皆符合國內黃麴毒素之法規限量標準(10 ppb)。

Aflatoxins are toxic and carcinogenic metabolic by-products produced primarily by Aspergillus flavusan and Aspergillus parasiticus. Growth of these fungi on certain foods may result in aflatoxin production and causing serious illness in humans. Aflatoxins contamination in crops mainly in peanut, maize, cereals, and spices is a worldwide food safety concern and has gained national attentions This study aims to develop a method for determination of aflatoxins in spices. Samples of spices including pepper, chili, and curry powder were extracted with 80% methanol and then analyzed by high performance liquid chromatography (HPLC). The LC separation was performed using Waters Cosmosil 5C18-AR column with the isocratic mobile phase consisting of 45% methanol. The flow rate was set at 1 mL/min. The current method demonstrated good linearity with regression coefficient over 0.995. The method was selective and offered good recoveries of various aflatoxins ranging from 61.85 to 101.34% (n=3). The coefficient of variance was below 12.05%. The limit of quantitation were from 0.5 (AFB_2 and AFG_2) to 1 ppb (AFB_1 and AFG_1). Four Aflatoxins could be simultaneously analyzed in a single 20- min run. This method was suitable for routine aflatoxins analysis in spices. In a survey of 33 commercial spices, AFB_1 was detected in 2 curry and 4 chili samples ranging from 0.79 to 2.74 ppb which were all below the regulatory limit (10 ppb) in Taiwan.