離子層析脈衝安培法對醣類之分析研究

Study of Carbohydrate by Ion Chromatography with Pulsed Amperometric Detection

孫亞君(Ya-Chun Sun)

指導教授 ： 黃悉雅

中原大學/理學院/化學研究所/碩士(2015年)

https://doi.org/10.6840/cycu201500943

摘要

本研究使用高效陰離子交換層析-脈衝安培流檢測法High-Performance Anion-Exchange Chromatography with Pulsed Amperometric Detection(HPAEC-PAD)對於單醣和雙醣進行分析，醣類在高鹼性(pH12-14)的環境中易於離子化，利用每一種醣分子有不同的解離常數(pKa)，調整流洗液的鹼性濃度，利用陰離子交換樹脂有不同的滯留時間可分離不同的醣分子，再利用脈衝安培流檢測法(Pulsed Amperometric Detection) (PAD)的氧化電位偵測濃度，並判斷分析物成分。
由於脈衝安培偵測器的靈敏度與鹼性環境密切相關，當鹼性溶液濃度增加時，醣類的解離度增加，解離常數小的醣類，需要在流洗液中加入比較強的流洗離子增加靈敏度，醣類組成複雜的樣品，如同時含有單醣、雙醣以及含有組成結構相似的多種醣樣品的分離則需要採用梯度流洗方式。所以當使用較低鹼性流洗液進行梯度流洗方式分離時，可在管柱後加入一較高濃度鹼性溶液即柱後反應(post-column reaction,PCR)以得到較佳的靈敏度，可同時分離多種醣類和得到最佳的靈敏度，並降低偵測極限。並針對實際樣品分析（如牛奶、優酪乳和奶粉），以乳糖和半乳糖分析結果之回收率、再現性作為此研究方法之確認。

關鍵字

高效陰離子交換層析 ； 脈衝安培流檢測法 ； 柱後反應

並列摘要

This study utilizes the “High-Performance Anion-Exchange Chromatography with Pulsed Amperometric Detection (HPAEC-PAD)” to analyze the monosaccharides and disaccharides. In high alkaline (pH12-14) environment, the carbohydrates were easily ionized. Since carbohydrates have different pKa, the concentration of alkaline eluent was investigated. The result showed that changing the concentration of alkaline eluent changes the retention time and could separate different carbohydrates.
The carbohydrate with lower pKa was washed with high ion concentration of NaOH to achieve higher sensitivity. For complicated carbohydrate samples, monosaccharides, disaccharides or similar structure carbohydrate was separated by gradient washing using NaOH. The result showed that the addition of NaOH could separate various carbohydrates with good sensitivity and detection limit. The proposed method was successfully applied in milk and yogurt samples and found the presence of lactose and galactose.

並列關鍵字

HPAEC ； PAD ； PCR

參考文獻

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