研究生: |
謝敏琦 Min-Chi Hsieh |
---|---|
論文名稱: |
毛細管微胞電泳層析掃集濃縮法結合77 K低溫螢光光譜分析對紅葡萄酒中微量逆轉醇即時光譜的鑑定與研究 On-line identification of trace trans-resveratrol in red wine using a sweeping technique combined with capillary electrophoresis/77 K fluorescence spectroscopy |
指導教授: |
林震煌
Lin, Cheng-Huang |
學位類別: |
碩士 Master |
系所名稱: |
化學系 Department of Chemistry |
論文出版年: | 2004 |
畢業學年度: | 92 |
語文別: | 中文 |
論文頁數: | 95 |
中文關鍵詞: | 毛細管微胞 、電泳層析 、77 K低溫 、螢光光譜 |
論文種類: | 學術論文 |
相關次數: | 點閱:131 下載:4 |
分享至: |
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本研究完成首次結合線上濃縮和毛細管電泳層析低溫螢光光譜偵
測法,並同時確認紅葡萄酒中逆轉醇77 K低溫的光譜。
利用sweeping-MEKC毛細管電泳層析法進行線上濃縮並分離逆轉
醇,當逆轉醇到達偵測窗口,瞬時將溫度降至77 K,測量on-line螢光光
譜,並和標準品的77 K低溫螢光光譜作指紋比對。
偵測紅酒萃取物中的逆轉醇最佳分離條件為matrix 20 mM
H3PO4,分離緩衝溶液內含30 mM H3PO4、150 mM SDS,溶液中甲醇
和水的比例為25:75 (v/v)。和normal-MEKC毛細管電泳層析法相比較,
使用sweeping-MEKC的毛細管電泳層析法,可以在偵測靈敏度上增加
1500倍。最低偵測極限是5 ppb (S/N = 3),偵測條件為激發螢光波長313
± 8 nm,吸收螢光波長400 ± 16 nm,使用濾光片濾掉320 nm以下的光
線。
利用毛細管微胞電泳層析濃縮掃集,結合on-line 77 K 螢光光譜偵
測法,不只可在紅酒萃取物中分離並偵測逆轉醇,且77 K低溫的線上
螢光光譜,更可確認逆轉醇在紅葡萄酒中的真實存在。
The feasibility of combining the techniques of on-line concentration
and capillary electrophoresis/low temperature fluorescence spectroscopy
(CE/LTFS) in the detection and identification of trans-resveratrol in red
wine at 77 K is demonstrated for the first time.
This technique, involving sweeping-micellar electrokinetic chromatog
-raphy (sweeping-MEKC), was used for the initial on-line concentration
and separation, after which, a cryogenic molecular fluorescence experiment
was performed at 77 K.
The complete, optimal separation of trans-resveratrol from a red wine
extract (in a sample matrix (an aqueous 20 mM H3PO4 solution)) could be
achieved with phosphate buffer(30 mM) containing SDS (150 mM) in a
methanol -water solution (25:75 v/v) , and a 1500-fold improvement in
detection sensitivity was obtained compared with the normal-MEKC
method. The limit of detection is 5 ppb (LOD). (S/N = 3). The excitation
wavelength was 313 ± 8 nm; the emission was measured at 400 ± 16
nm(with 320 nm cut filter).
The proposed method permits, not only the separation and detection
of trans-resveratrol from red wine extracts, but also ensures that the on-line
spectrum is readily distinguishable and can be un-ambiguously assigned at
77 K.
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