酚-硫酸法是多糖研究中最常用的总糖含量分析方法之一。然而，发现苯酚-硫酸法得到的结果普遍低于实际总糖含量，尤其是当多糖样品中含有酸性单糖时。因此，本研究的重点是揭示所提出的问题。通过对最佳吸收波长、线性范围、显色反应时间和温度等比色条件的优化，表明苯酚-硫酸法是一种方便、准确的总糖含量分析方法。此外，系统分析了10种常见单糖的显色能力，得到了每种单糖相对于葡萄糖的相对校正因子，证明这是总糖含量检测出现偏差的主要原因。并提出了苯酚-硫酸法应用过程中的关键点。本研究为苯酚-硫酸法准确测定总糖含量提供了科学的理论依据和可靠的实验研究方法，也将促进该简便方法在多糖研究中的应用。提出了苯酚-硫酸法应用中的要点。本研究为苯酚-硫酸法准确测定总糖含量提供了科学的理论依据和可靠的实验研究方法，也将促进该简便方法在多糖研究中的应用。提出了苯酚-硫酸法应用中的要点。本研究为苯酚-硫酸法准确测定总糖含量提供了科学的理论依据和可靠的实验研究方法，也将促进该简便方法在多糖研究中的应用。

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Phenol-sulfuric acid method is one of the most common methods applied to the analysis of total sugar content during polysaccharides study. However, it was found that the results obtained from the phenol-sulfuric acid method was generally lower than the real total sugar content, especially when acidic monosaccharides were contained in the polysaccharides samples. Therefore, the present study focused to unveil the proposed problem. Based on the optimization of colorimetric conditions, such as optimal wave length of absorption, linearity range, color reaction time and temperature, it indicated that the phenol-sulfuric acid method was a convenient and accurate way for the total sugar content analysis. In addition, the color-rendering capabilities of 10 common monosaccharides were systematically analyzed to obtain a relative correction factor for each monosaccharide relative to glucose, which was proved to be the main reason for the deviation in the detection of total sugar content. Moreover, the key points during the application of phenol-sulfuric acid method were suggested. This study provides a scientific theoretical basis and a reliable experimental research method for the accurate determination of total sugar content by the phenol-sulfuric acid method, and which will also promote the application of this convenient method in the polysaccharides study.