臺灣博碩士論文加值系統

摘要

由於目前市面果汁只標示具有維生素C的總量，並未註明其中L-ascorbic acid和isoascorbic acid個別含量。本研究建立一快速及準確之方法，能同時對L-ascorbic acid和isoascorbic acid分別做有效的定性及定量。
本實驗採用高效能液相層析法分析L-ascorbic acid和isoascorbic acid，並自行製備氧化態標準品和分析其純度。
在氧化態維生素C標準品的製備過程中，L-ascorbic acid需經過24小時，而和isoascorbic acid則需12小時方能完全氧化。所製備之標準品純度可高達約94~95%。因加入了凍結乾燥之步驟，而使得製備程序更加簡便，也使得標準品更加容易貯藏。
本實驗使用逆相C18層析管柱，以0.15％(v/v）醋酸水溶液作為移動相，以紫外光偵測器於245 nm下偵測，可同時分析還原態及氧化態之L-ascorbic acid和isoascorbic acid。

Abstract

Juices sold on the market are only labeled the total amount of ascorbic acid, not individual content of L-ascorbic acid and isoascorbic acid. The research is to establish a rapid and accurate method to simultaneously identify and quantitate both the L-ascorbic acid and isoascorbic acid in the juice.
The experiment was performed by analyzing L-ascorbic acid and isoascorbic acid with a high-performance liquid chromatograph. The standards of the two oxidized forms of ascorbic acids were prepared by ourselves. Their purities were also determined.
In preparing the standards of the oxidized forms of L-ascorbic acid and isoascorbic acid, it took 24 and 12 hours for preparing dehydroascorbic acid and dehydroisoascorbic acid, respectively. Purities of the prepared oxidized standards were high enough to reach 94~95%. The standards were freeze-dried. They were relatively easy to handle and store.
A C18 column, a mobile phase of 0.15% (v/v) acetic acid solution, and a ultraviolet detector of which the detection was made at 245 nm were used in the chromatographic system. The reduced forms and oxidized forms of both L-ascorbic acid and isoascorbic acid were simultaneously detected.

目錄
中文摘要 I
英文摘要 II
目錄 III
表目錄 VI
圖目錄 VII
第一章 前言 1
第二章 文獻整理 2
一、維生素C之起源與來源 2
(一) 維生素C之起源 2
(二) 維生素C之來源 2
二、維生素C之代謝與生理功能 3
(一) 維生素C之代謝 3
(二) 維生素C之生理功能 3
1. 刺激膠原蛋白的合成 3
2. 預防壞血病 4
3. 清除自由基 4
4. 抑制黑色素的形成 4
5. 增加亞鐵離子的吸收，與銅產生拮抗作用 4
三、維生素C之結構與物化性質 5
(一) 維生素C之結構 5
(二) 維生素C之物化性質 5
四、維生素C之應用 6
五、維生素C之分析 6
(一) 維生素C之萃取 6
1. 以萃取溶劑去除干擾物質 6
1-1. 維持酸性環境、能夠螯合金屬離子 7
1-2. 維生素C氧化酶活性的抑制和蛋白質去除 7
1-3. 澱粉的去除 7
1-4. 色素的去除 7
2. 物理方法之配合 7
3. 偵測條件之考量 8
3-1. 完全還原 8
3-2. 完全氧化 8
(二) 早期維生素C之分析 8
1. DCIP之滴定法 8
2. DNPH之比色法 9
3. 螢光衍生化法 9
4. Paper chromatography 10
5. Thin layer chromatography 10
(三) 近代維生素C之分析 11
1. Gas chromatography(GC) 11
1-1. GC - flame ionization detector(FID) 11
1-2. GC - mass(MS) 12
2. High-performance liquid chromatoghraphy(HPLC) 12
2-1. HPLC - refractive index detector(RI) 12
2-2. HPLC - ultraviolet detector(UV) 13
2-3. HPLC - flurometric detector 13
2-4. HPLC - electrochemical detector(EC) 14
3. Capillary zone electrophoresis(CZE) 14
六、實驗目的 15
第三章 材料與方法 16
一、材料 16
(一) 樣品 16
(二) 標準品 16
(三) 藥品 16
(四) 層析管柱 17
(五) 儀器 17
二、方法 18
(一) L-ascorbic acid及isoascorbic acid之分析 18
1. 標準品溶液之配製 18
2. 樣品溶液之處理 19
3. 層析條件 19
(二) DHAA和DHIAA之分析 20
1. LAA和IAA氧化程度之分析 20
1-1. LAA和IAA之氧化 20
1-2. LAA和IAA氧化程度之分析 20
2. DHAA和DHIAA標準品之製備 21
3. 製備所得之DHAA和DHIAA標準品之分析 22
3-1. HPLC法 22
3-2. 純度之分析 23
第四章 結果與討論 25
一、L-ascorbic acid及isoascorbic acid之分離 25
(一) L-ascorbic acid及isoascorbic acid標準品之分離 25
(二) 分析樣品中L-ascorbic acid及isoascorbic acid 27
二、 DHAA和DHIAA之分析 28
(一) DHAA和DHIAA之製備 28
(二) DHAA和DHIAA標準品之分析 29
1. HPLC法 29
2. 純度之分析 31
第五章 結論 32
表 33
圖 37
參考文獻 66

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