臺灣博碩士論文加值系統

肉品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，一個可適用於快速常規檢驗且可信之分析方法用於檢驗肉類食品之來源是必須的。鴕鳥肉由於具有高蛋白、高鐵、高鋅以及低膽固醇、低熱量、低脂肪的三高三低特性，成為符合健康飲食觀念的理想肉類。不肖商人常藉機以其他較便宜品種之肉類取代或參雜於鴕鳥肉中以降低成本、提高利潤。本論文之目標為利用高效能液相層析搭配電化學偵測系統將鴕鳥肉與雞肉、牛肉以及豬肉進行區別。結果顯示利用簡單的前處理步驟萃取肉汁後，便可於9分鐘內將四種肉成功區分，其中鴕鳥肉與雞肉具有牛肉及豬肉所缺乏之特異性波峰。此外，本方法並可以以極佳之敏感度及特異性區別不同口感及價位之鴕鳥肉等級。在室溫下存放之肉品，其主要波峰面積下降速率為在冷藏時之3倍。100 ℃隔水加熱5分鐘後所得之層析圖譜與加熱前僅有波峰面積上之差異，而無數量之改變，顯示本方法在肉品分級及熟肉樣品應用上之潛力。以酸性溶液、蛋白質沉澱溶液以及消化酵素等做樣品之前處理，可破壞、消化分析物，達到選擇性樣品純化之目的，可提供主要波峰性質探討之進一步資訊。未來，再進一步利用質譜儀鑑定層析圖譜中主要波峰之結構以及利用不同的樣品前處理方式，純化出具種別特異性之波峰，確認可區分肉品種別之標記之成分。

Identification of the origin of meat species is relevant to consumers for several reasons, either economical or religious. Thus, reliable and sensitive analytical tools are required for routine detection and identification of food from animal origin. Ostrich meat presents characteristic nutritional properties such as high protein content, rich in iron, zinc, and low in calorie, fat and cholesterol. The high unit-price and insufficient supply has made ostrich meat a target for adulteration and replacement by other cheap meat species. The purpose of the study was to validate high performance liquid chromatography method with electrochemical detection （HPLC-EC） for ostrich meat differentiation. 5 gram of meat mixed with 5 ml of 40 mM phosphate buffer, pH7.8, were homogenized for 1 min, and filtered before HPLC-EC analysis using copper nanoparticle plated electrodes. Ostrich meat could be differentiated from chicken, pork, and beef in 9 minutes by 4 major chromatographic peaks. An avian-specific peak at 3.5 minutes was identified for differentiation from mammal species. Storage of meat at various temperature and time could change the chromatographic pattern with the main peak reduction 3 times faster in room temperature （25 ℃） than in the refrigerator （4 ℃）. Different parts of ostrich meat showed distinct peak ratio that are characteristic for different meat grades. Acid and enzyme hydrolysis could uncover species-specific peaks and characteristic chromatographic pattern to facilitate the power of species differentiation. In conclusion, this HPLC-EC method appeared to be simple and suitable for rapid differentiation of ostrich meat from other three meats. The possibility of producing species-specific peaks by different chemical treatments warrant further study.

中文摘要 …………………………………………………………………I
英文摘要…………………………………………………………………II
目次 ……………………………………………………………………III
表次………………………………………………………………………VI
圖次 ……………………………………………………………………VII
第一章、前言……………………………………………………… 1
第二章、文獻探討………………………………………………… 4
第一節、鴕鳥肉的物理性質………………………………… 4
第二節、鴕鳥肉的營養價值………………………………… 6
第三節、肉品種別鑑定之重要性…………………………… 9
一、公共衛生…………………………………………… 9
二、消費者需求及權益………………………………… 9
第四節、肉品種別鑑定之方法介紹………………………… 10
一、分子生物學法……………………………………… 10
二、酵素免疫法………………………………………… 14
三、色層分析法………………………………………… 16
四、感官圖譜分析法…………………………………… 22
第五節、電化學搭配高效能液相層析（HPLC-EC）之原理及應 用24
一、電化學簡介………………………………………… 24
二、固定電壓技術……………………………………… 25
（一） 循環伏安法……………………………… 25
（二） 安培法 ………………………………… 27
（三） 流動注入分析法 ………………………… 28
三、固定電流技術……………………………………… 30
四、偵測胺基酸類之工作電極選擇…………………… 30
第三章、材料與方法……………………………………………… 33
第一節、實驗材……………………………………………… 33
一、肉類樣品來源……………………………………… 33
二、實驗儀器與樣品…………………………………… 33
(一)高效能液相層析系統………………………… 32
（二） 流動式注入分析系統 …………………… 34
（三） 電化學分析儀 …………………………… 34
（四） 其他儀器 ………………………………… 35
（五） 實驗藥品 ………………………………… 36
第二節、實驗目的及方法…………………………………… 37
一、以奈米銅網版印刷電極配合HPLC-EC偵測肉品之標準方法37
(一) 肉類樣品之前處理………………………… 37
(二) 移動相之製備……………………………… 37
(三) HPLC-EC分析之基本條件 ………………… 38
二、肌肉部位對層析圖譜之影響……………………… 38
三、室溫及冷藏對層析圖譜之影響…………………… 38
四、加熱對層析圖譜之影響…………………………… 39
五、不同前處理方式對層析圖譜之影響……………… 39
(一) 鹽酸（Hydrochloric acid, HCl）處理 39
(二) 三氯醋酸（Trichloroacetic acid, TCA）處理 40
(三) 木瓜酵素（Papain）處理 ……………… 40
(四) 胃蛋白酶（Pepsin）處理 ……………… 40
六、統計分析…………………………………………… 41

第四章、實驗結果 …………………………………………………42
第一節、以奈米銅網版印刷電極配合HPLC-EC偵測肉品之標準方法…42
第二節、肌肉部位對層析圖譜之影響 ………………………45
第三節、室溫及冷藏對層析圖譜之影響 ………………… 50
一、室溫25 ℃下存放72小時 ………………………… 50
二、4 ℃冷藏存放9天………………………………… 53
三、室溫及冷藏對於層析圖譜波峰面積變化之比較… 56
第四節、加熱對層析圖譜之影響…………………………… 59
第五節、不同前處理方式對層析圖譜之影響……………… 60
一、鹽酸（Hydrochloric acid, HCl）處理………… 60
二、三氯醋酸（Trichloroacetic acid, TCA）處理 61
三、木瓜酵素（Papain）處理………………………… 62
四、胃蛋白酶（Pepsin）處理………………………… 63
第五章、討論……………………………………………………… 64
第一節、以奈米銅網版印刷電極配合HPLC-EC偵測肉品之標準方法…64
第二節、層析圖譜中波峰成分之推測……………………… 66
第三節、肌肉部位對層析圖譜之影響……………………… 71
第四節、肉品降解對於層析圖譜之影響…………………… 73
第五節、不同處理方式對於層析圖譜之影響……………… 76
第六節、未來發展與展望…………………………………… 80
一、特異性波峰之成分結構鑑定……………………… 80
二、感官圖譜於氣相層析之應用……………………… 80
三、化學RFLP之假說…………………………………… 81
第六章、附錄……………………………………………………… 82
第七章、參考文獻………………………………………………… 89

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