以免疫親和性管柱配合螢光判讀機及高效液相層析儀檢測酒中之黃麴毒素   全文下載

Determination of Aflatoxins in Liquor Products by Immuno-affinity Column with Fluorometry and HPLC

酒為發酵食品之精華，但其農產品原料若貯藏不良，有受黴菌污染產生黃麴毒素之可能，又東方釀酒過程中常使用麴菌類，而部分麴菌菌種會產生黃麴毒素，若其原料或製程控制不當則產品中是否會有黃麴毒素污染是一相當值得研究的問題。本研究以黃麴毒素B1、B2、G1及G2之免疫親和性管柱萃取，再配以螢光判讀機或高效液相層析儀二種方法定量之；結果在酒中添加5或10ppb之黃麴毒素B1、B2、G1或G2時，不論使用何種定量法所測得之四種黃麴毒素回收率並無顯著差異，VICAM螢光判讀法對四種黃麴毒素之偵測極限皆為1 ppb，高效液相層析法對黃麴毒素B1、B2及G2之偵測極限皆為0.5ppb，但對黃麴毒素G1則為1 ppb。雖然螢光判讀法十分速簡適於大量檢體之篩檢，惟因無法區分各種黃麴毒素之各別含量，且最低檢測限量為1 ppb，故以螢光判讀法進行酒類檢體篩檢時將處理檢體量增為二倍，對於有螢光反應之檢體再以高效液相層析法確認，結果於民國八十四年七月至八十五年六月所抽驗之28件國產酒及72件大陸酒中，皆未檢出黃麴毒素。 Liquors are the spirits of fermentive products. The oriental fermentation processes use Aspergillus species extensively. However, a few Aspergillus species, especially Aspergillus flavus and A. parasiticus, can produce aflatoxins. Besides, the raw materials for liquor productions are liable to fungal pollution under improper storage conditions. There may have the possibility of aflatoxin contaminaion in the brewing product if the fermentive raw materials are moldy or the fermentation processes are polluted. An immuno-affinity column of aflatoxins B1, B2, G1 and G2 was used for sample extraction. Subsequently, either a fluorometry determination method or an HPLC analysis with Cosmosil 5C18-AR column was applied for aflatoxin detection. There was no significant difference between the recoveries of these two detection methods when analyzed by ANOVA (a=0.05). The detection limits of all the four aflatoxins were 1 ppb for the fluorometry method. The detection limits of HPLC analysis were 1 ppb for aflatoxin G1 and 0.5 ppb for the others. The fluorometry method is simpler and faster than HPLC analysis, especially when there are many samples to be screened. However, the fluorometry method cannot identify the amount of individual aflatoxin and has higher detection limits than HPLC analysis. Therefore, fluorometry screening requires double amount of sample to compensate for the lower sensitivity of this method. Those fluorescent- positive samples should also be further identified and quantified by HPLC analysis. Using these standard procedures, no aflatoxins were detected in a total of 100 samples obtained between July 1995 and June 1996. These samples comprised 28 domestic liquor products from the Taiwan Tobacco and Wine Monopoly Bureau and 72 liquor products from Mainland China."

Liquors are the spirits of fermentive products. The oriental fermentation processes use Aspergillus species extensively. However, a few Aspergillus species, especially Aspergillus flavus and A. parasiticus, can produce aflatoxins. Besides, the raw materials for liquor productions are liable to fungal pollution under improper storage conditions. There may have the possibility of aflatoxin contaminaion in the brewing product if the fermentive raw materials are moldy or the fermentation processes are polluted. An immuno-affinity column of aflatoxins B1, B2, G1 and G2 was used for sample extraction. Subsequently, either a fluorometry determination method or an HPLC analysis with Cosmosil 5C18-AR column was applied for aflatoxin detection. There was no significant difference between the recoveries of these two detection methods when analyzed by ANOVA (a=0.05). The detection limits of all the four aflatoxins were 1 ppb for the fluorometry method. The detection limits of HPLC analysis were 1 ppb for aflatoxin G1 and 0.5 ppb for the others. The fluorometry method is simpler and faster than HPLC analysis, especially when there are many samples to be screened. However, the fluorometry method cannot identify the amount of individual aflatoxin and has higher detection limits than HPLC analysis. Therefore, fluorometry screening requires double amount of sample to compensate for the lower sensitivity of this method. Those fluorescent- positive samples should also be further identified and quantified by HPLC analysis. Using these standard procedures, no aflatoxins were detected in a total of 100 samples obtained between July 1995 and June 1996. These samples comprised 28 domestic liquor products from the Taiwan Tobacco and Wine Monopoly Bureau and 72 liquor products from Mainland China.