Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/13059
標題: 以高效能液相層析結合奈米銅電極鑑定肉品種別之研究
Meat Species Identification by HPLC with Copper Nanoparticle Plated Electrode
作者: 李國閔
Lee, Kuo-Min
關鍵字: Meat species;肉品種別;HPLC;Copper Electrode;層析;銅電極
出版社: 獸醫學系
摘要: 
摘要
肉品的種別鑑定在公共衛生以及消費者權益上,具有相當重要的角色。不僅有利於政府對肉品衛生安全的管理,也保障了消費者的需求。因此,肉品種別鑑定及摻雜檢測方法的開發日遽增多,如電泳法、免疫分析法、色層分析法、DNA雜合反應及聚合酶鏈鎖反應等。但作為常規檢查的價值仍不實際。本實驗室利用奈米銅電極結合高效能液相層析,發展出一簡單、快速且經濟的方法。不需複雜的有機溶劑萃取步驟,即可依據種別特異之層析譜於短暫的十五分鐘內,鑑定出十六種人類經常食用的肉品。其中包含了生牛肉、豬肉、馬肉、羊肉、鹿肉、雞肉、鴨肉、鴕鳥、鱈魚、鮭魚、吳郭魚、鱷魚、干貝、牛蛙、蝦及蟹等。同一樣品或同種動物不同個體間所得到之波峰滯留時間差異度皆小於3%。將牛肉、豬肉及雞肉放置於室溫二十四小時及兩個冷凍-解凍過程之層析圖發現只有波峰的濃度改變,而不影響波峰譜之內容,若經由加熱100℃持續5分鐘處理後所得之層析譜亦僅有濃度之變化,此波峰比例於評估肉品新鮮度及加工肉品之鑑定具有極大之潛力。相同分類動物中雖然具有某些獨特之波峰,但尚無法定義其所代表之群體特異性或種別特異性,然而這些資訊於鑑別肉品混合物時,有助於篩選分類。在鴨及豬肉中,不同部位之肉品對於層析譜之影響非常微小,因此,此法具有相當大的潛力可作為一般肉品種別鑑定之工具並可應用於罕見肉品及法醫學上的檢測。如以酸、熱及酵素做樣品之前處理,可破壞或消化分析物,提供主要波峰性質探討之資訊以及相似層析譜間區別。未來,應可再進一步鑑別層析圖中主要波峰之結構,並深入的探討可能影響本分析方法之因子,如性別、品種、年齡、部位、加熱及加工等。若能將建立多種動物之層析譜資料庫,則可提供常規的檢測及配合政府防疫檢疫相關措施之參考,極具價值。

Abstract
Identifying the origin of meat species represents a considerable problem for food analysts and those seeking to comply with religious regulations. Consumers demand quality products that are honestly-labeled to assure meat safety and fair pricing. Therefore, there has been a need for a fast and routinely applicable meat species identification system. Here we have developed a rapid, inexpensive, and reliable technique which combines high-performance liquid chromatography and the electrochemical detection (HPLC-EC) with coppernanoparticle plated electrodes to identify 15 different meat species that commonly consumed by humans. Raw meats of 15 animal species including cattle, pigs, horses, goats, deer, chicken, duck, ostrich, cod, salmon, tilapia, alligator, scallop, bullfrog, shrimp and crab were identified by their specific chromatographic profiles in 15 minutes. Nine of the profiles exhibited a three-peak pattern, four (cattle, goat, pig and duck) exhibited a four-peak pattern and two species (horse and scallop) exhibited a two-peak pattern. The coefficient of variation of peak retention times were all less then 5.9% across repeated runs, geographical locations (leg and breast of duck, round and flank of pig) of the meat sources and among different subjects of the same species. This method does not require any organic extraction procedures and does not need derivatization for amino acid detection. When beef, pork, and chicken were exposed at room temperature for 24 hours or after 2 freeze-thaw cycles, only quantitative changes in peak area were found. Heating of these three meat samples at 100 ℃ for 5 minutes revealed similar quantitative changes in peak area, indicating the ratios between major peaks are likely applicable to cooked meats and are feasible for assessments of meat freshness. Chromatographical differences in geographical locations of the duck and pork were insignificant. Mixing of pork, beef and horse meat at 1 to 1 ratio were clearly identified by the developed method. Suggesting the method has great potential of being a routinely applicable identification tool for common and rare meat species with implications in forensic toxicology. Pretreatments of meat samples with acid, heat, and enzyme were studied to understand the nature of the major peaks and to further distinguish similar chromatographic profiles. Further studies are warranted to identify the structure of major peaks in profile, and to investigate factors that might affect the profile, specifically, the sex, breeds, ages, geographical locations of the animal and processing of meat. In conclusion, a simple, rapid and reliable HPLC-EC method was developed for identification of meat origins; this method is especially suitable for routine application and can uncover improper meat adulterations.
URI: http://hdl.handle.net/11455/13059
Appears in Collections:獸醫學系所

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