利用MALDI-TOF質譜法進行大量、快速篩檢Fumonisins毒素之偵測技術

The Development of High Throughput Screening Technique for Fumonisins by Matrix-Assisted Laser Desorption/Inoization Time-of-Flight Mass Spectrometry

林靜芸(Jing-yun Lin)

指導教授 ： 陳政男

朝陽科技大學/理工學院/應用化學系碩士在職專班/碩士(2010年)

摘要

伏馬鐮孢毒素（Fumonisins，FB）是串珠鐮刀菌（Fusarium moniliforme）所產生的二次代謝產物，主要汙染作物為玉米及其相關產品，會造成豬隻出現類似感冒症狀，使的投藥錯誤，造成巨大損失。不僅如此伏馬鐮孢毒素與人類的食道癌也極為相關。
伏馬鐮孢毒素中以FB1的毒素最強且量最大（70%），目前衛生署公告的穀物中FB1的分析方法為：使用一次性免疫親合管柱（NT$≧300元，成本較高）進行萃取且須經衍生反應後再以HPLC-FLD偵測。但此方法對於沒被汙染的農作物樣品中仍然要支出同樣高的成本，並不符合綠色環保的概念。因此，在這篇論文中，我們應用基質輔助雷射脫附游離/飛行時間質譜法（MALDI-TOF MS）開發出一個具有低成本，高靈敏度且可大量篩選FB1毒素的分析方法來輔助官方方法的不足。
本方法中的前處理只需要以甲醇和水萃取分析物後離心過濾即可以MALDI-TOF/MS來進行偵測。當只以α-CHCA為基質時，只能在穀物（玉米）中測到1 ppm的Fumonisins，因此不具可行性，但在添加Cs+ 後可增加分析靈敏度1000倍，偵測極限達到1 ppb。此法所需成本低（NT≦10）且偵測時間小於1分鐘，可節省不必要的材料及時間的浪費，相當符合當前綠色化學的環保概念。

關鍵字

伏馬鐮孢毒素 ； 基質輔助雷射脫附游離飛行時間質譜法

並列摘要

Fumonisins are secondary metabolites of Fusarium moniliforme, mainly in corn and corn-based products. It may cause flu-like symptoms on pig and result in wrong treatment. Moreover, fumonisins are associated with human esophageal cancer.
Fumonisin B1 (FB1) is the most toxic and most natural abundant type (70%) in Fumonisins. The official analysis method for FB1 is using disposable immunoaffinity column (NT$≧300) for extraction then following derivatization before HPLC-FLD measurement. However, the cost of the specific column and the time consuming are not good for high throughput analysis, especially for the non-contaminated samples. If there is a low cost and high throughput screening methods to filter out most uncontaminated samples, a lot of time consuming and cost would be avoided. Thus, we develop a low-cost, high sensitivity screening method for FB1 by MALDI-TOF MS to achieve this goal.
In this method, the pretreatment just need solvent (MeOH/H2O) extraction following centrifugation, then detected by MALDI-TOF/MS using α-CHCA as the matrix and Cs+ to increase sensitivity to 1 ppb. The detection limit would be only 1 ppm without Cs+. The cost of sample preparation for each sample is less than NT$ 10.0 dollars and analysis time for each sample is only 1 min or less. This is in line with current environmental protection concept of green chemistry.

並列關鍵字

Fumonisins ； MALDI-TOF MS

參考文獻

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