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

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

胡瓈文(Li-Wen Hu)

指導教授 ： 陳政男

朝陽科技大學/理工學院/應用化學系/碩士(2014年)

摘要

橘黴素(Citrinin，CIT)為紅麴菌發酵在生成紅色素時產生的代謝產物，是一種具有腎臟及肝臟及致畸胎毒性的真菌毒素。因為現在市面上具有養生概念之紅麴相關產品越來越多，導致其中可能含有橘黴素的風險也隨之提高。因此，本論文應用基質輔助雷射脫附游離/飛行時間質譜儀(MALDI-TOF MS)開發出具有低成本，且可快速篩選CIT毒素的分析方法。
本方法中的前處理只需要以約2 mL的溶劑萃取分析物後離心，取上清液即可以MALDI-TOF/MS來進行偵測。以α-CHCA為基質時，標準品的偵測極限為1 ppm，在基質中添加α-環糊精後，提高10倍偵測極限到100 ppb。在真實檢體小麥及紅麴薄餅的偵測極限可達200 ppb；在真實檢體紅麴米及紅麴膠囊的偵測極限可達1 ppm。此法所需成本很低（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

基質輔助雷射脫附游離飛行時間質譜法 ； 橘黴素

並列摘要

Citrinin (Citrinin, CIT) , a kind of mycotoxins with teratogenic toxicity and harmful to kidney and liver, is a metabolite of Monascus fermentation during generating red pigment. Since more and more health-related Monascus fermentation products are introduced to the market, the risk of citrinin contaiminations may increase. Therefore , I develope a low-cost, and rapid screening method to detect CIT by using matrix-assisted laser desorption ionization / time of flight mass spectrometry (MALDI-TOF MS) .
On this method, the pretreatment process requires only about 2 mL solvent to extract the sample centrifuged, the supernatant can be detected directly by MALDI-TOF/MS. When using α-CHCA as matrix, the detection limit is 1 ppm for standard, after adding α-cyclodextrin in the matrix, the limitation of detection can be improved 10 times to the 100 ppb. In the real specimen of wheat and red yeast pancake the detection limit can reach to 200 ppb ; for red yeast rice and red yeast capsules the detection limit can reach to 1 ppm. This method requires very low cost (NT ≦ 10) for each sample and the detection time is less than one minute. By this method, I may avoid unnecessary waste of materials and time consuming. This is in line with current environmental protection concept of green chemistry.

並列關鍵字

Citrinin ； MALDI-TOF MS

參考文獻

2. 廖家鼎、 林柏全、 林旭陽、 闕麗卿、 施養志，「市售含紅麴之食品中橘黴素含量調查」食品藥物研究年報， 第ㄧ期 ， 第109-116頁(2010)。

1. Xu, B. J., Jia, X. Q., Gu, L. J., Sung, C. K., “Review on the qualitative and quantitative analysis of the mycotoxin citrinin,” Food Control, Vol. 17, pp. 271-285(2006).

7. Wild, D., T’oth, G., Humpf, H. U., “New Monascus Metabolite Isolated from Red Yeast Rice(Angkak, Red Koji),” Journal of Agricultural and Food Chemistry, Vol. 50, pp. 3999-4002(2002).

8. Shu, P. Y., Lin, C. H., “Simple and Sensitive Determination of Citrinin in Monascus by GC–Selected Ion Monitoring Mass Spectrometry,” Analytical Sciences, Vol. 18, pp. 283-287(2002).

9. Li, Y., Zhou, Y. C., Yang, M. H., Yang, Z. O.,“Natural occurrence of citrinin in widely consumed traditional Chinese food red yeast rice, medicinal plants and their related products,” Food Chemistry, Vol. 132, pp. 1040-1045(2012).