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<p><i>Cuscuta reflexa</i> Roxb. has long been used in traditional medical systems in India and other nations against a wide range of illnesses. This paper relates to the development and validation of a high-performance thin-layer chromatography (HPTLC)‒densitometric method for the simultaneous estimation of quercetin and gallic acid in <i>C. reflexa</i> (whole plant) and the correlation of their content to its antioxidant activity determined by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. Ascending development was carried out on precoated silica gel 60 F<sub>254</sub> plates using toluene‒ethyl acetate‒formic acid (40:55:5, <i>V/V</i>) as the mobile phase. Quercetin and gallic acid appeared as compact bands at <i>R</i><sub>F</sub> 0.56 ± 0.02 and 0.38 ± 0.01, respectively on densitometric scanning at 385 and 285&#xa0;nm. Their contents in <i>C. reflexa</i> extract were found to be 1.31 ± 0.11 and 0.75 ± 0.02 mg/g, respectively, on a dry weight basis. The method was validated in accordance with the International Council for Harmonization (ICH) guidelines and was found to be robust, accurate, and precise. Total phenolic and flavonoid contents were found to be 79.1 ± 1.6 and 117.9 ± 2.5 mg gallic acid equivalent/g and 98.3 ± 0.9 and 157.8 ± 1.4 mg quercetin equivalent/g in methanol extract and ethyl acetate fraction of <i>C. reflexa</i>, respectively. The extract and fraction of <i>C. reflexa</i> successfully suppressed free radicals with a half maximal inhibitory concentration&#xa0;(IC<sub>50</sub>) value of ~50&#xa0;µg/mL and was compared with ascorbic acid (IC<sub>50</sub> ~4.7&#xa0;µg/mL). The antioxidant activity of <i>C. reflexa</i> showed a direct correlation to its total phenol and flavonoid contents vis-à-vis gallic acid and quercetin.</p>

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A validated high-performance thin-layer chromatography method for the simultaneous quantification of quercetin and gallic acid in Cuscuta reflexa Roxb. and correlating their content to its in vitro antioxidant activity

  • 期刊论文

  • 发表日期：

    2025年9月29日

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  • Akram Choudhary,
  • Zakiya Usmani,
  • Mohd Shafeeque,
  • Nisha Vats,
  • Showkat R. Mir,
  • M. Shahar Yar

摘要

Cuscuta reflexa Roxb. has long been used in traditional medical systems in India and other nations against a wide range of illnesses. This paper relates to the development and validation of a high-performance thin-layer chromatography (HPTLC)‒densitometric method for the simultaneous estimation of quercetin and gallic acid in C. reflexa (whole plant) and the correlation of their content to its antioxidant activity determined by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. Ascending development was carried out on precoated silica gel 60 F254 plates using toluene‒ethyl acetate‒formic acid (40:55:5, V/V) as the mobile phase. Quercetin and gallic acid appeared as compact bands at RF 0.56 ± 0.02 and 0.38 ± 0.01, respectively on densitometric scanning at 385 and 285 nm. Their contents in C. reflexa extract were found to be 1.31 ± 0.11 and 0.75 ± 0.02 mg/g, respectively, on a dry weight basis. The method was validated in accordance with the International Council for Harmonization (ICH) guidelines and was found to be robust, accurate, and precise. Total phenolic and flavonoid contents were found to be 79.1 ± 1.6 and 117.9 ± 2.5 mg gallic acid equivalent/g and 98.3 ± 0.9 and 157.8 ± 1.4 mg quercetin equivalent/g in methanol extract and ethyl acetate fraction of C. reflexa, respectively. The extract and fraction of C. reflexa successfully suppressed free radicals with a half maximal inhibitory concentration (IC50) value of ~50 µg/mL and was compared with ascorbic acid (IC50 ~4.7 µg/mL). The antioxidant activity of C. reflexa showed a direct correlation to its total phenol and flavonoid contents vis-à-vis gallic acid and quercetin.