醣類成分之分析研究

The study of saccharides analysis

李泳平(Yung-Ping Lee)

指導教授 ： 李水盛

國立臺灣大學/醫學院/藥學研究所/碩士(2004年)

https://doi.org/10.6342/NTU.2004.01738

摘要

本研究是針對植物中所含某些之醣類成份進行定性與定量分析，嘗試建立出一套方便的分析方法，適用於不同醣類，包括單醣、醣醇和寡醣，策略為進行簡易之過乙醯化反應，找尋適當之逆相高效能液相層析分析條件及檢測法。

研究發現下列高效液相層析法分析條件可在30分鐘內分析過乙醯化之單醣、雙醣、三醣、四醣和五醣類分子。: Lichrospher® 100 RP-18e (4 x 250mm, 5μm)為固定相，移動相為0-10分鐘30%體積比之氰甲烷與水，10-30分鐘以每分鐘2.5%線性增加氰甲烷至80%體積比，流速為1.0mL/min，樣品注射量為20 μL，管柱溫度35℃，分別以紫外光檢測波長210nm，和蒸發光散射偵測器(ELSD)檢測，漂流管(drift tube)的溫度35℃、噴霧氣體壓力2.8 bar。

相較於紫外光偵測，蒸發光散射偵測有幾個優點，如較不易受到高紫外光吸收的物質干擾，且適用於梯度沖提等，故蒸發光散射偵測較適合此分析方法。此方法使用的層析條件及有機移動相能應用於LC-NMR串聯技術，及進行大量醣類成份的分離。

關鍵字

蒸發光散射偵測 ； 高效能液相層析分析 ； 醣類

並列摘要

This study was aimed to analyze the content of saccharides in plants both qualitatively and quantitatively. We tried to develop a facile analytical method for different saccharides, including monosaccharides, polyols, and oligosaccharides. The strategy for such purpose was composed of a simple peracetylation reaction and development of an HPLC analytic condition including detection tool.

The HPLC conditions for analyzing peracetyl saccharides developed are as follows: a Lichrospher® 100 RP-18e column (4 x 250mm, 5μm) as stationary phase, MeCN-H2O (30:70, v/v, 0-10 min; to 80:20, 10-30 min, linear gradient) as mobile phase, flow rate 1.0 mL/min, injection volume 20 μL, and column temperature 35℃, detection at UV 210 nm or by evaporative light scattering detector (ELSD), setting drift tube 35℃ and nebulizer gas 2.8 bar. The peracetyl saccharides including mono-, di-, tri-, tetra-, and penta-saccharide were well resolved and analyzed within 30 min under such conditions.

In comparison with the UV detector, ELSD has some advantages in quantitative analysis, such as less interference by high UV absorbance substances and compatible to gradient elution. Therefore, ELSD is more suitable than UV detector in this study. The developed LC system (ELSD) can be hyphenated to SPE-NMR via a splitter and can be used in a large scale separation of such peracetyl saccharides.

並列關鍵字

saccharides ； ELSD ； HPLC

參考文獻

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