食品中棒麴毒素之檢驗方法探討   全文下載

Optimization of Analytical Method for Patulin in Foods

棒麴毒素存在於多種發黴的水果、蔬菜及穀物中，尤以蘋果及其加工製品最常受到棒麴毒素污染。衛生福利部於「食品中污染物質及毒素衛生標準」訂有蘋果製品、飲料類及嬰幼兒食品中棒麴毒素之限量標準，亦公告「食品中黴菌毒素檢驗方法－－棒麴毒素之檢驗（MOHWT0005.02）」對應衛生標準執行檢驗。惟有實驗室反映現行公告檢驗方法於前處理添加果膠酶會影響回收率，且部分檢體於光譜分析時因基質干擾無法判讀數據。本研究針對上述問題優化前處理流程及評估以液相層析串聯質譜儀分析棒麴毒素之可行性。優化內容包括減少取樣量、刪除果膠酶及減壓濃縮步驟、調整以25%乙腈溶液作為回溶液及採用同位素內標校正流程，並改以液相層析串聯質譜儀分析。本研究依衛生標準挑選不含棒麴毒素之蘋果汁、蘋果醋、黑棗汁及綜合果泥等4件不同類型之空白檢體，分別添加5及50μg/kg棒麴毒素依優化之檢驗方法執行添加回收試驗，結果顯示，同日內之平均回收率介於96.1-110.5%，CV為1.2-6.6%，異日間之平均回收率介於97.0-112.0%，CV為2.3-6.6%，測試結果皆符合「食品化學檢驗方法之確效規範」，定量極限均為5μg/kg。優化後之方法可節省2-4小時之檢驗時間，且方法之準確度及精密度皆有所提升。以本研究優化之方法進行市售產品之方法適用性驗證，17件檢體皆為未檢出棒麴毒素，符合衛生標準。後續將依研究成果進行現行檢驗方法修訂及公告，供各界參考使用。

Patulin has been found in many kinds of fruits, vegetables and grains, especially apple and apple products. The Ministry of Health and Welfare in Taiwan announced the“Sanitation Standard for Contaminants and Toxins in Food”to limit the maximum level of patulin in apple products, beverage, and food for infant and young child. The“Method of Test for Mycotoxins in Foods-Test of Patulin (MOHWT0005.02)”was also announced for tests corresponding with the sanitation standards. However, it was reflected that the pretreatment with pectinase in the current method would affect the recovery of patulin. Besides, matrix interference occurred during spectral analysis for some samples. The aim of this research was to solve the above problems and to evaluate the feasibility of liquid chromatography/tandem mass spectrometry (LC-MS/MS) analysis. The optimized conditions included: reducing the sampling size, omitting the pectinase and vacuum concentration steps, using 25% acetonitrile to dissolve the residue, using an isotope-labeled internal standard of patulin to calibrate the data and analyzing by LCMS/ MS. The recovery was tested by spiking patulin at 5 and 50μg/kg into four types of blank matrices without patulin, including apple juice, apple cider vinegar, black date juice and mixed puree, which were chosen according to the sanitation standards. The average recoveries ranged from 96.1 to 110.5%, with the coefficients of variance between 1.2 and 6.6% in intra-day analysis; the average recoveries ranged from 97.0 to 112.0%, with the coefficients of variance between 2.3 and 6.6% in inter-day analysis. The results all fulfilled the«Validation Guideline of the Food Chemical Testing Method”published by the TFDA. The limits of quantification of patulin were all 5μg/kg. The optimized method could save 2 to 4 hours of experimental time, with improved accuracy and precision. The optimized method was employed to verify the applicability of marketed products. None of a total of 17 samples was tested to contain patulin, which is in accordance with the sanitation standards. The current method will be further revised and announced based on this study for public use.