- 期刊
食品中下痢性貝毒檢驗方法之建立
Simultaneous Quantification of Diarrhetic Shellfish Poisoning Toxins in Bivalve Molluscs Using LC-MS/MS
摘要
下痢性貝毒(diarrhetic shellfish poisoning toxins, DSP)是由有毒的藻類所產生的化合物，經貝類攝食後而蓄積在體內造成污染，主要污染之食品有文蛤、牡蠣、孔雀蛤、扇貝等貝類。本研究以甲醇萃取孔雀蛤及牡蠣檢體，經離心、過濾後，以液相層析串聯質譜儀(liquid chromatograph/tandem mass spectrometer, LC/MS/MS)檢測，可同步分析下痢性貝毒中較常見之okadaic acid (OA)、dinophysistoxin-1 (DTX-1)及gymnodimine-A (GYM-A)共3種毒素。離子源採電灑游離法(electrospray ionization,ESI)，OA及DTX-1採負離子模式，GYM-A採正離子模式。層析條件以含6.7 mM氨水的90%乙腈溶液與去離子水為移動相進行梯度層析分離，流速為0.2 mL/min，層析管柱為Waters X-bridge C18，注入量為20 μL。研究結果顯示，3種貝毒之基質匹配檢量線相關係數皆大於0.995；以孔雀蛤及牡蠣為基質，分別添加80、160、240 ppb各毒素標準品，其平均回收率介於81-114%之間(n=3)，變異係數皆小於17.07%。本方法可在20分鐘內同步分析3種下痢性貝毒，相較於傳統小鼠生物試驗法，具有省時、方便、不需犧牲動物、可同步定量分析多種毒素等優點，可實際應用於食品中下痢性貝毒之檢測。
並列摘要
Diarrhetic shellfish poisoning toxins (DSP) are produced by algae and can accumulate in filter feeding bivalves. A method to determine the lipophilic marine biotoxins from extracts of different species of bivalve molluscs was developed. Okadaic acid (OA), dinophysistoxin-1 (DTX-1) and gymnodimine-A (GYM-A) were determined. Mussel and Oyster were extracted with methanol and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with an electrospray (ESI) ionization interface. Using the positive and negative ion mode, the parent ion and product ions of each diarrhetic shellfish poisoning toxin were selected. The LC separation was performed using a Waters X-bridge C18 column with the mobile phase consisting of gradient program of water and 90% acetonitrile with 6.7mM ammonia, at a flow rate of 0.2 mL/min. Regression coefficient for matrix-match calibration curve has to be at least 0.995. Good recoveries were found among various diarrhetic shellfish poisoning toxins, ranging from 81 to 114% (n=3). The coefficient of variance was below 17%, indicating this method is reliable. Three diarrhetic shellfish poisoning toxins could be simultaneously analyzed in a single 20-min run. This method provided higher sensitivity and accuracy than the mouse assay and did not require to sacrify animals. This method could be applied for routine DSP analysis.
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