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

High Throughput Screening for Zearalenone by Matrix-Assisted Laser Desorption/Inoization Time-of -Flight Mass Spectrometry

林曉珮(Shiau-Pei 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(2010年)

摘要

玉米赤黴烯酮 (Zearalenone，ZEA) 毒素是黴菌毒素 (Mycotoxin) 中的一種，乃真菌的二次代謝產物。常見於受污染之玉米、大小麥、燕麥。
目前穀物的保存技術較以往進步，受黴菌毒素污染的農作物比例相對減少。台灣官方衛生署公告穀物中ZEA的分析方法，必須使用一次性的免疫親合管柱(immunoaffinity column)對黴菌毒素進行濃縮後再以HPLC分離及定量，然而對於大部分未受污染的農作物樣品的檢驗，仍需支出同樣高成本及時間，並不符合經濟效益。若需要大量的樣品檢測，以衛生署公告的方法常造成不必要的資源浪費，因此開發低成本、快速、靈敏的可靠初步篩檢方法先行偵測出未受污染的穀物，則可避免不必要的浪費。
在本篇論文中，我們運用MALDI-TOF MS本身具有高靈敏度、並可大量且快速的樣品分析之優點開發具有低成本，且可大量初步篩檢ZEA毒素之分析方法。在本方法中，以矽藻土去除基質的樣品製備方法既簡單且成本低廉，由實驗結果發現以鋁箔為輔助偵測ZEA毒素時可得到比其它基質更好的偵測效果，其圖譜在低質量範圍背景雜訊干擾少，展現出良好的均勻性。使用此方法的樣品製備成本低於台幣10元，平均分析一個樣品也的時間只要1分鐘左右，偵測極限則可低至 10 ppb，與官方公告方法之偵測極限相當。

關鍵字

基質輔助雷射脫附游離飛行時間質譜法 ； 玉米赤黴烯酮

並列摘要

Zearalenone (ZEA) is one of the mycotoxins which are the secondary metabolites of fungi. It is found in corn, wheat, barley and oats.
In Taiwan, the official analysis method for ZEA is using disposable immunoaffinity column coupling with high performance liquid chromatography (HPLC) which has the detection limit of 10 ppb. However, the modern preservation of grain has reduced the contamination of ZEA in corps. The cost of the specific column and the time consuming are not good for high throughput analysis. 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.
Here we report a low cost and high throughput screening method for the analysis of ZEA using matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). This method includes two parts. First, the sample preparation using celite to remove sample matrix is simple and low cost. Second, without general matrix for co-crystal with analyte, aluminum foil is used to obtain low noise analyte spectra in low mass range. 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, detection limit can reach to 10 ppb which is comparable with the official method.

並列關鍵字

MALDI-TOF MS ； Zearalenone

參考文獻

1. Petra Blechova, Pavla Havlova, Dagmar Gajdosova, Josef Havel1, “New Possibilities of Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry to Analyze Barley Malt Quality. Highly Sensitive Detection of Mycotoxins,” Environmental Toxicology., pp. 403-408 (2006).

2. Berthiller, F., Schuhmacher, R., Buttinger, G., Krska, R., “Rapid simultaneous determination of major type A- and B-trichothecenes as well as zearalenone in maize by high performance liquid chromatography-tandem mass spectrometry,” J Chromatogr A., Vol. 1062, pp. 209-216 (2005).

3. Michael Sulyok, Franz Berthiller, Rudolf Krska, Rainer Schuhmacher, “Development and validation of a liquid chromatography/tandem mass spectrometric method for the determination of 39 mycotoxins in wheat and maize,” Rapid Commun. Mass Spectrom., Vol. 20, pp. 2649-2659 (2006).

6. Zdenka Brenn-Struckhofova, Margit Cichna-Markl, Christina Bo1hm, Ebrahim Razzazi-Fazeli, “Selective Sample Cleanup by Reusable Sol-Gel Immunoaffinity Columns for Determination of Deoxynivalenol in Food and Feed Samples,” Anal. Chem., Vol. 79, pp. 710-717 (2007).

9. David G. Schmale., and Gary P. Munkvold., “Mycotoxins in Crops: A Threat to Human and Domestic Animal Health” The Plant Health Instructor., DOI: 10.1094 (2009).