Carbohydrate analysis by a phenol-sulfuric acid method in microplate format
Carbohydrate analysis by a phenol-sulfuric acid method in microplate format
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DOI：
10.1016/j.ab.2004.12.001
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发表时间：
2005-04-01
影响因子：
2.9
通讯作者：
Lee, YC
中科院分区：
文献类型：
--
作者：
Masuko, T;Minami, A;Lee, YC
在碳水化合物分析的众多比色方法中，苯酚-硫酸法是最简单、最可靠的方法。它已用于测量寡糖、蛋白聚糖、糖蛋白和糖脂中的中性糖。该方法因其灵敏性和简单性而被广泛使用。在其原始形式下，它需要 50-450 nmol 的单糖或等效物进行分析，因此不足以用于珍贵的样品。之前报道过仅需要 10-80 nmol 糖的缩小版本。我们现在对该方法进行了修改和优化，以使用 96 孔微孔板实现高通量、获得更高的灵敏度并节省试剂。这种改进和优化的方法可实现更长的线性范围（对于人类为 1-150 nmol）和出色的灵敏度。而且，我们的方法更方便，不需要摇动，也不需要遮盖，只需不到 15 分钟即可完成。该方法的速度和简单性使其最适合分析大量样品，例如色谱馏分。 (c) 2004 Elsevier Inc. 保留所有权利。
Among many colorimetric methods for carbohydrate analysis, the phenol-sulfuric acid method is the easiest and most reliable method. It has been used for measuring neutral sugars in oligosaccharides, proteoglycans, glycoproteins, and glycolipids. This method is used widely because of its sensitivity and simplicity. In its original form, it required 50-450 nmol of monosaccharides or equivalent for analysis and thus is inadequate for precious samples. A scaled-down version requiring only 10-80 nmol of sugars was reported previously. We have now modified and optimized this method to use 96-well microplates for high throughput, to gain greater sensitivity, and to economize the reagents. This modified and optimized method allows longer linear range (1-150 nmol for Man) and excellent sensitivity. Moreover, our method is more convenient, requiring neither shaking nor covering, and takes less than 15 min to complete. The speed and simplicity of this method would make it most suitable for analyses of large numbers of samples such as chromatographic fractions. (c) 2004 Elsevier Inc. All rights reserved.