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<p>The phenol-sulfuric acid method is the simplest and most frequently used procedure to determine the carbohydrate concentration. This method was developed to quantify purified or partially purified carbohydrates at concentrations below 0.1&#xa0;g L⁻¹. However, its application in fermentation processes is limited due to the inherent characteristics of biological samples, which typically contain a variety of metabolites, high concentrations of macromolecules, and residual substrates. These factors can interfere with the quantification process. Thus, a modified phenol-sulfuric acid method was developed to control the reaction temperature, improving measurement accuracy and minimizing the impact of interferences. The results indicate that temperature, the phenol concentration, and reagent stability are the key parameters affecting absorbance measurement. The modified method involves a single premixed reagent in the reaction mixture instead of the conventional use of phenol and sulfuric acid. This approach allows for accurate and reliable measurement of monosaccharides, disaccharides, and polysaccharides with a concentration 20-fold higher (up to 1.75&#xa0;g L⁻¹) than the conventional phenol-sulfuric acid method. For that, the proposed method involves the reduction of the dilutions in the biological samples, thereby enhancing the repeatability, reproducibility, and trueness of the experimental results.</p>

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A robust method for quantification of total carbohydrates in complex biological samples: an alternative to the phenol-sulfuric acid method

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月7日

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  • Luis Eduardo García-Martínez,
  • Ernesto Favela-Torres,
  • Ruberth Rivera Pérez,
  • Christian Jesús Mora-Pérez

摘要

The phenol-sulfuric acid method is the simplest and most frequently used procedure to determine the carbohydrate concentration. This method was developed to quantify purified or partially purified carbohydrates at concentrations below 0.1 g L⁻¹. However, its application in fermentation processes is limited due to the inherent characteristics of biological samples, which typically contain a variety of metabolites, high concentrations of macromolecules, and residual substrates. These factors can interfere with the quantification process. Thus, a modified phenol-sulfuric acid method was developed to control the reaction temperature, improving measurement accuracy and minimizing the impact of interferences. The results indicate that temperature, the phenol concentration, and reagent stability are the key parameters affecting absorbance measurement. The modified method involves a single premixed reagent in the reaction mixture instead of the conventional use of phenol and sulfuric acid. This approach allows for accurate and reliable measurement of monosaccharides, disaccharides, and polysaccharides with a concentration 20-fold higher (up to 1.75 g L⁻¹) than the conventional phenol-sulfuric acid method. For that, the proposed method involves the reduction of the dilutions in the biological samples, thereby enhancing the repeatability, reproducibility, and trueness of the experimental results.