タウリン-過ヨウ素酸試薬を用いた高速液体クロマトグラフィーによる糖類の蛍光分析

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書誌事項

タイトル別名
  • Fluorometry of saccharides by HPLC using taurine-periodate reagent.
  • タウリン‐過よう素酸試薬を用いた高速液体クロマトグラフィーによる糖類の蛍光分析
  • タウリン カ ヨウソサン シヤク オ モチイタ コウソク エキタイ クロマトグ

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説明

A highly sensitive method for fluorometric detection and determination of reducing and nonreducing saccharides by HPLC was developed using taurine-periodate reagent by heating in a neutral solution. Moreover, nonreducing oligosaccharides and glycosides gave enhanced fluorescence under a mild optimum condition.The mixture of reducing and nonreducing saccharides was separated with 0.50.7mol dm-3 borate (pH 8.7) as mobile phase through an anion exchange Aminex A-27 or ISA07/S2504 column. The eluate was added with 640mol m-3 taurine-2mol m-3 sodium metaperiodate solution and heated at 140°C and the fluorescence intensity was measured with a detector. Standard curves for reducing and nonreducing saccharides were linear in the range from 0.27 nmol/10μl to 26.7 nmol/ 10μl Detection limit (S/N=3) of D-sorbitol was 0.13 nmol/10μl. On the other hand, the separation of nonreducing oligosaccharides and glycosides was carried out with 0.1mol dm-3 borate (pH 7) as mobile phase on a Hitachi gel 3013N column. The postcolumn fluorescence reagent consisted of 100mol m-3 taurine-6 mol m-3 sodium metaperiodate (pH 6.8 with K2B4O7) was added. After mixing, the eluate was heated at 130°C to give fluorescence. Standard curves for α-methyl-D-glucoside, sucrose, raffinose and stachyose were linear in the wide range from 0.1 nmol/10μl to 75 nmol/10μl. Detection limit of α-methyl-D-glucoside was 60 pmol/10μl and those of sucrose, raffinose and stachyose 30 pmol/10μl. The relative fluorescence in tensities of these saccharides increased with the increase in the length of an oilgosaccharide chain.

収録刊行物

  • 分析化学

    分析化学 35 (10), 869-874, 1986

    公益社団法人 日本分析化学会

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