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Surveillance and identification of Zeranol-immunoreactive compounds in moldy rice in Taiwan
|關鍵字:||zeranol;雙偵測;HPLC;dual detection;moldy rice;高效能液相層析;黴菌毒素;電化學;發霉米||出版社:||獸醫學系暨研究所||引用:||呂鋒洲。黴菌毒素（譯述）。中正書局，台北市，台灣，1982。 胡啟章。電化學原理與方法。五南出版社，台北，台灣，2007。 孫逸民、沈玉舜、趙敏勳、謝明學、劉興鍵。儀器分析。全威圖書有限公司。台北縣。台灣。2006。 師維孝。中興大學獸醫系碩士學位論文。發展以電化學偵測Zeranol及分析動情素相關基因於台灣地區乳癌之表現，2006。 陳銘華。中興大學化學系碩士學位論文。液相層析質譜術於食物中黃麴毒素及赭麴毒素之檢測，2005。 黃振文、孫守恭、馬慧英。台灣產鐮胞菌之研究。中菌會刊。7:1-22, 1992。 賴玉艷。中興大學化學系碩士學位論文。利用化學修飾電極同步偵測次黃嘌呤、黃嘌呤、尿酸之研究，1999。 Abdelhamid AM, Kelada IP, Ali MM, and El-Ayouty SA. Influence of zearalenone on some metabolic, physiological and pathological aspects of female rabbits at two different ages. Arch Tierernahr. 42: 63-70, 1992. Becci PJ, Johnson WD, Hess FG, Gallo MA, Parent RA, and Taylor JM. Combined two-generation reproduction-teratogenesis study of zearalenone in the rat. J Appl Toxicol. 2: 201-206, 1982. Berek L, Petri IB, Mesterhazy A, Teren J, Molnar J. Effects of mycotoxins on human immune functions in vitro. Toxicol In Vitro. 15: 25-30, 2001. Berg, H. 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Zeranol（Z）是一種具動情素活性的非類固醇類黴菌毒素（zearalenone , Zen）衍生物，為FDA核准使用於肉品工業的生長促進劑。而Zen為Fusarium屬所產生之黴菌毒素，此種黴菌可生長在發霉之燕麥、大麥、小麥、高粱及米上。近年來的研究指出，長期食用含有Z、Zen和其相關代謝產物（Zs）殘留的食物與乳癌的發生有關。因此，人類在進食發霉穀物的同時恐無法避免攝入Zs，尤其以米食為主的台灣人民暴露到Zs的量可能更多。本研究主要目的即在探討台灣本土米中Zs的含量且同時鑑定發霉米中具有Z免疫活性物質之結構。實驗計自台灣北、中、南、東區各連鎖超市及米店取得共221件様本，先以ELISA篩選出具Zs免疫活性之様本，再以高效能液相層析法予以鑑定。結果顯示，在所測的221個樣品中僅21個米樣品（9.5%）含有高於1 ppb之Zs免疫活性，且此殘留值遠低於FAO所制定之穀物中殘留標準50-1000 ppb。在所測的7個黃豆、3個玉米片樣品中則分別有4個（57.14%)及有3個（100%）高於1 ppb之Zs免疫活性。抽選特定樣品，以所發展之高效能液相層析配合紫外光及電化學偵測發現，所有抽撿樣品之Zs含量皆在偵測極限（10 ppb）以下。綜合以上結果，台灣新鮮米中之Zs殘留量極低。在Zs檢驗方法之開發上，我們以高效能液相層析同時串聯紫外光及電化學而完成雙偵測系統（HPLC-UV-EC）用以同時檢測Zs之存在；此法之UV偵測極限在10-45 ppb之間、電化學極限為25-90 ppb之間，且有良好的精確度及準確度。將上述方法應用在已發霉米之分析上發現，經過固相萃取（回收率80%左右）後，可見到疑似有Z和β-Zearalenol以及α-Zearalenol的波峰出現，其含量分別為18.74、8.15、3.39 ppm，顯示發霉米的確有食品安全上的疑慮。質譜鑑定具Zs免疫活性之成份初步發現為m/z 329、m/z 331、m/z 293及m/z 295之物質，其分子量與Zs或是黃麴毒素接近。綜合言之，本實驗除有助了解國內米中Zs的含量現況外，亦發展了一個適合常規檢驗並定量穀物中Zs的方法，且米類食品若保存不良而有發霉情形將造成食品安全上的疑慮。
Zeranol (Z) is a nonsteroidal agent with estrogenic activity that is used as a growth promoter in the cattle industry in the United States. Zeranol may also be formed in vivo from the metabolism of mycotoxin zearalenone (Zen) produced by Fusarium spp that grows on grains including maize, oat, barley, wheat, and sorghum. The residue of Z, Zen, and its metabolites (Zs) in food may exert estrogenic effects on mammalian endocrine system and contribute to the development of human breast cancer. Therefore, safe food free from Zs in meat and grains are important and reliable analytical methods are required to monitor Zs residue in food. The objectives of the study were to investigate the prevalence of Zs in rice products in Taiwan and to identify Z-immunoreactive compounds associated with mold production. A total of 221 rice samples were collected from north, middle, south and east regions of Taiwan over one-year period for screening of immunoreactive Zs activity by enzyme-linked immunosorbent assays (ELISA). The results indicated that only 21 rice samples (21/221=9.5%) have Z-immunoreactivity higher than 1 ng/g. This level is well bellow the FAO approval level of 50-1000 ppb in grains, suggesting that the contamination level of Zs in fresh rice in Taiwan was low. A simple and rapid high performance liquid chromatography (HPLC) method with electrochemical (EC) and UV (wavelength=274 nm) dual detection was validated for simultaneous determination of Zs in various matrices such as rice, soybean, and cereals after solid-phase extraction. Recoveries were around 80% and the limits of detection ranged from 10 ng/ml to 25 ng/ml for UV and from 50 ng/ml to 90 ng/ml for EC detection with good accuracy and reproducility. Suspected Z, β-Zearalenol and α-Zearalenol peaks were observed in moldy rice, which may implicate possible exposure of Zs to human health. Mass spectrometry analysis of suspected Z-immunoreactive compound revealed substrates with m/z 329, m/z 331, m/z 293, and m/z 295. This developed method was suitable for a routine analysis of Z-immunoreactive compounds in grains and can uncover contamination of Zs in rice.
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