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<p>Polysaccharide-rich plant exudate gums are increasingly explored as functional ingredients in food and pharmaceutical applications, necessitating reliable analytical methods for their characterization and quality control. In this study, a UV–Visible spectrophotometric phenol–sulfuric acid assay was optimized and validated for total carbohydrate quantification in <i>Bauhinia racemosa</i> exudate gum and its purified water-soluble (BRG-S) and insoluble (BRG-I) fractions. Sample purification was performed by ethanol precipitation, and protein and phenolic interferences were minimized by trichloroacetic acid treatment. Hydrolysis conditions were optimized using a One-Factor-at-a-Time (OFAT) approach, identifying 60&#xa0;°C, 0.10% (v/v) sulfuric acid, and 30&#xa0;min as optimal for maximum colorimetric response. Method validation was performed in accordance with ICH guidelines using D-glucose as the calibration standard. The assay exhibited excellent linearity over 0–100&#xa0;µg/mL (R² &gt; 0.995), high precision (relative standard deviation, %RSD) below 3%, and good accuracy (recoveries ranging from 96.39% to 104.68%). The limit of detection (LOD) and limit of quantification (LOQ) were 0.87&#xa0;µg/mL and 2.63&#xa0;µg/mL, respectively. Application of the validated method revealed carbohydrate contents ranging from 76.77% to 92.08% as glucose equivalents across different gum fractions. The optimized and validated assay provides a simple, sensitive, and reproducible quality-control tool for routine carbohydrate analysis of <i>Bauhinia racemosa</i> gum, supporting its standardization and sustainable valorization for food and pharmaceutical applications.</p> Graphical abstract <p></p>

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Validation of a UV–visible phenol–sulfuric acid method for carbohydrate quantification in Bauhinia racemosa gum

  • 期刊论文

  • 发表日期：

    2026年5月13日

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  • Chirantan Sandip Saigaonkar,
  • Sandhya R. Shewale,
  • Uday S. Annapure

摘要

Polysaccharide-rich plant exudate gums are increasingly explored as functional ingredients in food and pharmaceutical applications, necessitating reliable analytical methods for their characterization and quality control. In this study, a UV–Visible spectrophotometric phenol–sulfuric acid assay was optimized and validated for total carbohydrate quantification in Bauhinia racemosa exudate gum and its purified water-soluble (BRG-S) and insoluble (BRG-I) fractions. Sample purification was performed by ethanol precipitation, and protein and phenolic interferences were minimized by trichloroacetic acid treatment. Hydrolysis conditions were optimized using a One-Factor-at-a-Time (OFAT) approach, identifying 60 °C, 0.10% (v/v) sulfuric acid, and 30 min as optimal for maximum colorimetric response. Method validation was performed in accordance with ICH guidelines using D-glucose as the calibration standard. The assay exhibited excellent linearity over 0–100 µg/mL (R² > 0.995), high precision (relative standard deviation, %RSD) below 3%, and good accuracy (recoveries ranging from 96.39% to 104.68%). The limit of detection (LOD) and limit of quantification (LOQ) were 0.87 µg/mL and 2.63 µg/mL, respectively. Application of the validated method revealed carbohydrate contents ranging from 76.77% to 92.08% as glucose equivalents across different gum fractions. The optimized and validated assay provides a simple, sensitive, and reproducible quality-control tool for routine carbohydrate analysis of Bauhinia racemosa gum, supporting its standardization and sustainable valorization for food and pharmaceutical applications.

Graphical abstract