Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/13059
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dc.contributor.advisor周濟眾zh_TW
dc.contributor.advisorChi-Chung Chouen_US
dc.contributor.author李國閔zh_TW
dc.contributor.authorLee, Kuo-Minen_US
dc.date2005zh_TW
dc.date.accessioned2014-06-06T06:50:13Z-
dc.date.available2014-06-06T06:50:13Z-
dc.identifier.urihttp://hdl.handle.net/11455/13059-
dc.description.abstract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。zh_TW
dc.description.abstractAbstract 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.en_US
dc.description.tableofcontents目錄 目錄…………………………………………………………………………….I 表次……………………………………………………………………………V 圖次…………………………………………………………………………...VI 中文摘要……………………………………………………………………..VII 英文摘要……………………………………………………………………VIII 第一章 前言…………………………………………………………………...1 第二章 文獻探討……………………………………………………………...3 第一節 肉品種別鑑定之重要性………………………………………...3 一、消費者需求…………………………………………………..…3 二、市場價格公平性……………………………………………..…3 三、公共衛生……………………………………………………..…4 第二節 肉的組成………………………………………………………...4 一、外觀結構………………………………………………………..4 二、顯微結構………………………………………………………..5 三、主要成分………………………………………………………..7 (一)水及脂質…………………………………………….….8 (二)蛋白質……………………………………………..…..10 (三)碳水化合物………………………………………..…..14 (四)可溶性的非蛋白物質……………………………..…..15 第三節 現有檢測肉品種別之方法…………………………………….17 一、酵素免疫分析法………………………………………………17 二、分子生物學方法………………………………………………18 三、電泳分析法……………………………………………………20 四、色層分析法……………………………………………………21 第四節 電化學搭配高效能液相層析之原理及應用………………….22 一、電化學簡介……………………………………………………22 二、固定電壓技術…………………………………………..……..22 (一)循環伏安法…………………………………….………23 (二)安培法………………………………………………….23 (三)流動分析………………………………………….…....23 三、固定電流技術…………………………………………..…..…24 四、工作電極之選擇………………………………………………24 五、電化學搭配流動分析器………………………………………25 第三章 材料與方法…………………………………………………..……...26 第一節 實驗材料…………………………………………………..…...26 一、實驗儀器…………………………………………………….…26 (一)電化學分析儀………………………………………….26 (二)高效能液相層析儀…………………………………….26 (三)其他儀器……………………………………………….26 二、實驗藥品………………………………………………….……27 三、肉品樣品………………………………………………….…...28 第二節 實驗方法…………………………………………………..…...28 一、建立以奈米銅網版印刷電極配合HPLC(HPLC-EC)偵測肉品之標準方法………………………………………………..28 (一)樣品前處理…………………………………………….28 (二)磷酸緩衝溶液的配製………………………………….29 (三)HPLC-EC分析之基本條件……………………………29 二、電極再現性之探討…………………………………………..29 三、探討以HPLC-EC層析譜區別肉品種別之最佳條件………29 (一)移動相離子強度之影響…………………………..…..30 (二)移動相pH值之影響………………………………….30 (三)移動相中有機溶劑比例之影響……………………….30 四、探討不同樣品前處理之影響………………………………..30 (一)酸處理…………………………………..……………..31 (二)加熱處理………………………………………….…...31 (三)酵素消化處理……………………………..……...…...31 五、可能影響電化學層析譜之肉品變異因子的探討……….…..31 (一)低溫保存以及高溫對分析樣品之影響……………....31 (二)動物體不同部位肌肉之差異………………………....32 (三)肉品新鮮程度對層析譜之影響………………….…...32 (四)混合不同種別肉品對層析譜之影響………….….…..32 第四章 實驗結果………………………………………………………...…..33 第一節 分析方法之確效……………………………………………….33 一、電極再現性探討………………………………………………33 二、移動相條件之選擇……………………………………………35 (一)移動相中離子強度對分析物之影響………………….35 (二)移動相中pH值對分析物之影響……………………..35 (三)移動相中有機溶劑添加之比例對分析物之影響…….38 三、前處理方法之探討……………………………………………38 第二節 肉品種別鑑定層析圖………………………………………….41 一、各種肉品之HPLC-EC標準層析圖譜………………………..41 二、層析圖滯留時間變異評估……………………………………41 第三節 相關影響因子之探討………………………………………….48 一、肉品樣品反覆冷凍解凍對層析譜之影響……………………48 二、肉品樣品新鮮度對層析譜之影響……………………………48 三、肉品加熱對層析譜之影響………………………...………….48 四、不同部位肌肉對層析譜之影響……..………………………..53 五、混合不同肉品對層析譜之影響………………………………54 第五章 討論………………………………………………………………….55 第六章 未來可能之發展與應用…………………………………………….65 第一節 血液樣品在種別鑑定上之應用……………………………….65 第二節 毛髮組成應用於種別鑑定之潛力…………………………….68 第三節 調味烹飪煮熟樣品之分析……….……………………………70 參考文獻……………………………………………………………………...72zh_TW
dc.language.isoen_USzh_TW
dc.publisher獸醫學系zh_TW
dc.subjectMeat speciesen_US
dc.subject肉品種別zh_TW
dc.subjectHPLCen_US
dc.subjectCopper Electrodeen_US
dc.subject層析zh_TW
dc.subject銅電極zh_TW
dc.title以高效能液相層析結合奈米銅電極鑑定肉品種別之研究zh_TW
dc.titleMeat Species Identification by HPLC with Copper Nanoparticle Plated Electrodeen_US
dc.typeThesis and Dissertationzh_TW
item.openairetypeThesis and Dissertation-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.languageiso639-1en_US-
item.grantfulltextnone-
item.fulltextno fulltext-
item.cerifentitytypePublications-
Appears in Collections:獸醫學系所
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