以酵素免疫法檢測中藥材內赭麴毒素、伏馬毒素及T-2毒素污染之研究

Occurrence of Ochratoxin fumonisin and T-2 toxin in Chinese medicines by an enzymelinked immunosorbent assay method

李冠奕(li guan yi)

指導教授 ： 鄧正賢

亞洲大學/健康學院/生物科技學系碩士班/碩士(2007年)

摘要

赭麴毒素A ( oclirato 他llA , oTA ）是由麴黴蘭和青黴菌所產生的• 常分布污染糧食和動物飼料，是一種很強的腎毒性致癌物質並具有免疫特性，在許多國家中赭麴毒素A 是引起腎病的原因之一• 伏馬毒素（F 、mDnisills ）由F 鵬磁tun 菌所產生的，常見於農產品生長或儲存過程，目前懷疑與人類食道癌有關。T - Z 毒素主要是由Ftlsariulll 類的黴菌所產生，而T - 2 毒素會侵襲皮膚及呼吸道，亦為戰爭使用的黃雨化學武器，因此本實驗選譯對此三項黴菌毒素進行檢測。而本實驗之目的在於完整建立中藥材內緒麴毒素A 、伏馬毒素、T - 2 毒素的污染狀況，並建立中藥材內緒麴毒素A 、伏馬毒計Bl 、T －二毒素污染的酵素免疫分析檢測方法作LIS 勾，以完整進行一系P , J 黴菌毒素污染中藥材之研究”
實驗研究顯示中藥含有干擾酵素免疫分析檢測的內含物• 依照簡單的萃取方式會使檢測出現偽陽性，經過多次實驗後改用Acetol 血斑e 及因相純化管柱來做萃取，實驗才有較佳的精準性，另外實驗會繼續測試台灣11 地區的10 種辛香類中藥材，其中被三種黴菌毒素污染的含量，
結果顯示緒麵毒素檢出包括八角（11 / 11 ）、胡椒（11/11 ）、豐翹( 10 / 10 ）、義朮（11 / 11 ）、花椒（11 / 11 ）、鬱金（11 / 11 ）、薑黃（10 / 9 ）、T 香（10 / 9 ）、肉桂皮（11/11 ）及小茴香（11/10 ) ，含量分別依序為0 . 28 -23 . 00 、0 . 54 -14 . 24 、0 . 34 -15 . 92 、0 . 39 -18 . 02 、3 . 63 -16 . 78 、0 . 05 -17 . 23 、6 . 08 -31 . 15 、2 .52-25 . 43 、4 . 65 ，-49 . 64 、1 . 13 -23 . 93 一ug/kg T - 2 毒素檢出八角、胡椒、較朮、薑黃及丁香內有微量的污染。伏馬毒素則需要再深入探討檢驗方式。
辛香類中藥材為國人日常生活上所常用的食品配料，但是本實驗檢驗出多數含有OTA 的污染，這個發現是很令人震驚的事情，因此篡於安全因素及考量經濟效應，建議辛香類中藥材的管理及製備上宜須特別小心，並需要進一步多點採樣並重複篩選確定，進而才規範其限量標準。

關鍵字

酵素免疫分析、緒麴毒素、伏馬毒素、T - 2 毒素

並列摘要

Abstract
Mycotoxins are predominantly by-products of fungal metabolic processes. among which. aflatoxin ¡ochratoxins ¡Fumonisins ¡T-2 toxin and citrinin are the most commonly found ones in contaminated food. Mycotoxins have both antibiotic and toxic properties. the most important of which are its nephrotoxic. hepatitoxic. carcinogenic, and immunotoxic properties.
Ochratoxin A (OTA) is produced by several species of the flingal genera Aspergillus and Penicihliuni. These fungi are ubiquitous and the potential for the contamination of food stuffs and animal feed is widespread. OTA has both antibiotic and toxic properties. the most important of which are its nephrotoxic. carcinogenic, and inununotoxic properties. It has been the cause of a nephropathy in many countries.
Fumonisins (FBi) are toxins produced by Fusarium species that grow on several agricultural commodities in the field or during storage. Consumption of fumonisins can result in cancer of the esophagus.The trichotliecene mycotoxins are low molecular weight (250-500 dIaltons) nonvolatile compouiids produced by filamentous fungi (molds) of the genera Fusarium. Myrotecium. Trichodernia. Stachybotiys and others. T-2 toxins may enter the body through the skin and digestive or respiratory epithelium. The air attacks in Laos have been described as ¡§yellow rain¡¨ and consisted of a shower of sticky. yellow liquid that sounded like rain as it fell from the sky
The purpose of this study was to investigate the presence of OTA. Fuinonis ins and T-2 toxin in Chinese C¡¦nide Drugs in Taiwan We first established the methods of analysis of OTA. Fumonisins and T-2 toxin, then the samples of Chinese crude drugs were collected from traditional markets or retail shops and analyzed for the mycotoxins. Using organic solvents
xu
extraction, solid phase extraction clean up and EL ISA to analyze 10 kinds of Chinese crude drugs form 11 spaces in Taiwan.
The methods were applied to about 10 different samples. OTA was detected in 11 samples of Fructus Anisi Stellati. 11 samples of Fructus Piperis Nigri. 10 samples of Fnictus Amomi Rotundus. 11 samples of Rhozima Curcumae. 11 samples of Pericarpium Zaiithoxyli. 11 samples of Root tuber of aromatic tunneric. 9 samples of Rliizoma C¡¦urcumae Longae. 9 samples of Flos Caryophylli. 11 samples of Cortex Cinnamomi. 10 samples of Fructus Foeniculi. measurable at 0.28-23.00 ¡¥ 0.54-14.24
0.34-15.92 0.39-18.02 3.63-16.78 0.05-17.23 6.08-31.15 2.52-25.43
4.65-49.64 1.13-23.93 1.tglkg. T2 was detected in Fnictus Anisi Stellati. Fructus Piperis Nii, Rhorna Curcumae. Rhizoma Curcumae Longae and Flos Caryophylli but the contents of T2 are less. FBi was need discuss deeply.

並列關鍵字

ELISA Ochratoxins Fumonisins T-2 toxin

參考文獻

30. 陳曉錚，李衛宗，談國雄，闕麗卿，傅幼敏，施養志，應用二種管柱厚衍生化反應分析食品中黃麴毒素之比較，藥物食品檢驗局調查研究年報.24:421-430 2006.

1. Flannigan B.. Hui S.C.. The occur ance of aflatoxin-producing strains of Aspergillus fiavus fl tile mould floras of ground spice. 3. Appi. Bacteriol. 1976: 41: 411-418.

2. Tliirumaia-Devi K.. Mayo M.A.. Reddy Eddy K. Gopal . Tangni Einmauel K.. Occurence of ochratoxin A in black pepper. coriander. ainder and turmericin India. Food Additives and Contaminants. 2001:18(9): 830-835.

7. Van der Merwe. K.J., Steyn. P.S.. Fourie. L.. Scott, D.B.. Theron. J.J.. Ochratoxiu A. a toxic metabolite produced by Aspergillus ochraceus. Nature 1965 205(4976).

mycotoxins in food and feed in 2003.