臺灣博碩士論文加值系統

雞精中大多成份以小分子蛋白質及胺基酸等功能性物質居多，其中所含之肌肽 (carnosine)與甲肌肽 (anserine)主要存在於脊椎動物 (如雞、牛、豬和魚)的腦、骨骼肌與神經組織中，此類胜肽物質根據文獻指出具有抗氧化、調節免疫反應、降血脂及傷口癒合等功能，也被建議具有潛在的治療效果可用於影響阿茲海默疾病及帕金森氏疾病和相關疾病的發病機制。在台灣每年約有800萬隻淘汰雞，為了進一步增加雞骨(含肉)副產物的利用價值，本研究目的欲著重在不同加工雞精產品中肌肽與甲肌肽含量分析的方法建立，包括樣品前處理部分的固相萃取方法最佳化，並利用液相層析儀搭配光電二極體陣列偵測器 (LC-PDA)來偵測找出最佳層析條件。本實驗取雞骨(含肉)副產物作為樣品，以濕熱烹調法、乾熱烹調法及高溫高壓法進行雞精樣品製作，檢測這三種加工條件下產品中肌肽與甲肌肽之含量，並與市售雞精樣品比較。結果顯示當以移動相A/B混合成80%/20%為起始條件可得到肌肽與甲肌肽的最佳分離效果。此外，樣品前處理使用強陽離子交換的固相萃取匣有最佳的回收率，分別為101.90±12.28%和105.23±17.62%。自製樣品經濕熱烹調法分別得到肌肽與甲肌肽23.56±2.64與79.74±6.13 μg/ml，以乾熱烹調法則得到23.36±7.85與80.99±17.27 μg/ml，而以高溫高壓法則得到25.42±0.01與157.73±1.97 μg/ml之含量，三組加工條件並無顯著差異。與市售雞精樣品比較，市售滴雞精測得較高的肌肽與甲肌肽之含量，分別為232.35±5.57和674.53±17.40 μg/ml。肌肽與甲肌肽含量以BCA法分析的總蛋白質含量校正後，自製雞精產品中以乾熱烹調法製作可得含量較高的肌肽與甲肌肽，其含量分別為9.92和39.60 mg/g蛋白質，而市售雞精產品中以滴雞精肌肽與甲肌肽含量較高，分別為18.77和51.92 mg/g蛋白質，兩者皆以乾熱烹調之方法製作，因此以乾熱烹調之方法製作加上適當濃縮步驟將能製成較高濃度肌肽與甲肌肽的雞精產品。綜合上述，此分析條件可用於雞精中肌肽與甲肌肽含量分析，並可作為肉精製品生產品管之參考。

Chicken essence contains many functional compounds such as small-molecule proteins and amino acids. It contains dipeptides carnosine (CAR) and anserine (ANS) that are found in brain, skeletal muscles and nerve tissues of vertebrates such as chickens, cattle, pigs and fish. They have been shown in literature to have immuno-response modulation, blood fat reduction and wound healing functions. They also have been suggested to have potential therapeutic applications in individuals with Alzheimer's disease as well as potential effects against the pathogenesis of Parkinson’s disease and related disorders. In Taiwan there are about 8 million spent chickens for disposal, and in order to increase their value, we intended to develop an analytical method to analyze CAR and ANS in chicken essence products by using liquid chromatography coupled with photo diode array detector (LC-PDA) and solid-phase extraction (SPE) for sample clean up, to study the effect of different processing conditions on the levels of CAR and ANS and compare to other commercial products. This study used spent chicken bones to make essence products by wet heat processing, dry heat processing and high temperature high pressure processing methods. The results showed that, for normal-phase LC performance, the combination of 80%/20% for mobile phase A/B at initial stage of LC gave better separation of CAR and ANS. As for the SPE condition, using strong-cation exchange cartridge gave the highest recovery of CAR and ANS, which was 101.90±12.28% and 105.23±17.62%, respectively. Essence samples obtained from spent chicken bones showed no significant difference in either CAR or ANS content among these three processing conditions. To compare CAR and ANS concentrations in commercial dripped chicken essence samples, those made under dry heat had the highest CAR and ANS concentrations, which were 232.35±5.57 and 674.53±17.40 μg/ml, respectively. After adjusted by the protein content determined by BCA method, the CAR and ANS concentration in the dry heat processed chicken essence samples were the highest, which were 9.92 and 39.60 mg/g protein, respectively. This was also observed in the commercial products made by dry heat processing, which gave the highest CAR and ANS concetration of 18.77 and 51.92 mg/g protein, respectively. Therefore, dry heat processing method with proper concentrating step can be used to make chicken essence products with high levels of CAR and ANS. In conclusion, the analytical method for CAR and ANS was developed and capable of detecting chicken essence products. The result might provide an useful information for chicken essence production and for quality control purpose for meat essence products.

中文摘要 I
Abstract III
誌謝 V
縮寫對照表 VI
目錄 VII
圖目錄 X
表目錄 XII
第1章、 前言 1
第2章、 文獻回顧 2
2.1 雞精 2
2.1.1 雞精來源 2
2.1.2 一般雞精之生產/製作流程 2
2.1.3 滴雞精之生產/製作流程 3
2.1.4 熬雞精之生產/製作流程 3
2.2 雞精之營養成份 3
2.2.1 一般成份 3
2.2.2 二肽(Dipeptide) 9
2.2.3 肌肽(β-alanyl-L-histidine; carnosine) 9
2.2.3.1 肌肽之化學結構 9
2.2.3.2 肌肽之合成與代謝途徑 13
2.2.3.2.1 肌肽之合成 13
2.2.3.2.2 肌肽之吸收 13
2.2.3.2.3 肌肽之代謝 14
2.2.3.3 肌肽之機能性功效 16
2.2.3.3.1 肌肽與細胞抗衰老之代謝 16
2.2.3.3.2 肌肽與醣解作用的能量代謝 16
2.2.3.3.3 肌肽與蛋白酶之變化 18
2.2.3.4 肌肽的治療疾病潛力 18
2.2.3.5 肌肽與癌症(Cancer) 18
2.2.3.6 肌肽與阿茲海默症(Alzheimer’s disease) 19
2.2.3.7 肌肽與帕金森氏症(Parkinson’s disease) 21
2.2.3.8 肌肽與糖尿病相關疾病(Diabetes-related disease) 25
2.2.4 甲肌肽 (β-alanyl-3-methyl-L-histidine; Anserine) 26
2.2.4.1 甲肌肽之化學結構 26
2.2.4.2 甲肌肽之合成 26
2.3 肌肽與甲肌肽之分析方法 28
2.3.1 固相萃取法 (Solid-phase extraction, SPE) 28
2.3.2 液相層析法 ((Liquid chromatography, LC) 31
2.3.3 光電二極體陣列偵測器 (Photo diode array detector, PDA) 32
2.4 統計方法 32
2.4.1 階層族群分析法 (Hierarchical cluster analysis, HCA) 32
第3章、 研究方法 33
3.1 試驗目的 33
3.2 試驗設計 33
3.3 試驗藥品 33
3.4 試驗材料 35
3.5 試驗方法 37
3.5.1 雞精產品之製備 37
3.5.1.1 濕熱烹調法(wet heat processing) 37
3.5.1.2 乾熱烹調法(dry heat processing) 37
3.5.1.3 高溫高壓烹調法(high temperature high pressure processing) 37
3.5.2 固相萃取(Solid-phase extraction, SPE)方法 37
3.5.3 LC-PDA之層析條件 40
3.5.4 蛋白質濃度定量法 42
3.5.4.1 Bicinchoninic acid (BCA)法 42
3.5.4.2 福林-酚(Folin-lowry)法 42
3.5.5 統計方法 42
第4章、 結果與討論 43
4.1 以LC-PDA於不同移動相比例下分析肌肽和甲肌肽標準品之層析結果 43
4.2 以最佳LC-PDA法分析肌肽與甲肌肽含量之確效分析 44
4.2.1 肌肽與甲肌肽標準溶液之標準曲線 44
4.2.2 以LC-PDA方法定量肌肽與甲肌肽含量之再現性 44
4.2.3 以LC-PDA方法定量肌肽與甲肌肽含量之準確度 44
4.3 固相萃取結果 51
4.3.1 肌肽標準品經固相萃取分離之結果 51
4.3.2 甲肌肽標準品經固相萃取分離之結果 51
4.4 雞精產品中肌肽與甲肌肽之含量 54
4.5 總蛋白質含量 58
4.5.1 Bicinchoninic acid (BCA)方法之結果 58
4.5.2 福林-酚(Folin-Lowry)方法之結果 58
4.6 雞精產品中肌肽和甲肌肽含量以總蛋白質含量校正後之結果 63
第5章、 結論 70
作者簡介 72
參考文獻 73

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