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<p><i>Caesalpinia bonduc</i> is traditionally used in various pharmacological conditions. The various plant parts reflect the presence of various phytoconstituents. The aim of this study was to identify, quantify, and validate selected constituents of cassane diterpenes from <i>C. bonduc</i> leaf extract using thin-layer chromatography‒mass spectrometry<b> (</b>TLC‒MS). An extract of the dried leaf powder of <i>C. bonduc</i> was prepared using 60% aqueous methanol. The instrumental TLC method was performed to identify and quantify cassane diterpene at specific <i>R</i><sub>F</sub> values. The proposed method used aluminum plates precoated with silica gel 60 F<sub>254</sub> as the stationary phase and <i>n</i>-hexane‒ethyl acetate (6:4, <i>V</i>/<i>V</i>) as the mobile phase for separation purpose. TLC‒MS analysis confirmed the retention of β-caesalpin and α-caesalpin, which were analyzed and validated in compliance with the International Council for Harmonization (ICH) Q2 (R2) guidelines at a wavelength of 580&#xa0;nm after derivatization with anisaldehyde reagent. Various bands were separated on TLC plate, highlighting the presence of different phytoconstituents. The interpretation of the <i>m</i>/<i>z</i> ratio of the mass spectra of a selected TLC band revealed the presence of cassane diterpene β-caesalpin and α-caesalpin at <i>R</i><sub>F</sub> values of 0.64 and 0.78, respectively. For both caesalpins, a linear response was observed in the concentration range of 200–1200&#xa0;ng/band. In <i>C. bonduc</i>, the TLC‒MS method was successfully applied to identify β-caesalpin and α-caesalpin. For the determination of β- and α-caesalpin in the hydroalcoholic leaf extract of <i>C. bonduc</i>, the developed approach was revealed to be robust, precise, accurate, and specific.</p>

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Advanced thin-layer chromatography‒mass spectrometry validation and comprehensive analysis of bioactive phytochemicals in Caesalpinia bonduc leaf extract

  • 期刊论文

  • 发表日期：

    2025年4月5日

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  • Jinal N. Tandel,
  • Usmangani Chhalotiya,
  • Heta Kachhiya,
  • Devang Tandel

摘要

Caesalpinia bonduc is traditionally used in various pharmacological conditions. The various plant parts reflect the presence of various phytoconstituents. The aim of this study was to identify, quantify, and validate selected constituents of cassane diterpenes from C. bonduc leaf extract using thin-layer chromatography‒mass spectrometry (TLC‒MS). An extract of the dried leaf powder of C. bonduc was prepared using 60% aqueous methanol. The instrumental TLC method was performed to identify and quantify cassane diterpene at specific RF values. The proposed method used aluminum plates precoated with silica gel 60 F254 as the stationary phase and n-hexane‒ethyl acetate (6:4, V/V) as the mobile phase for separation purpose. TLC‒MS analysis confirmed the retention of β-caesalpin and α-caesalpin, which were analyzed and validated in compliance with the International Council for Harmonization (ICH) Q2 (R2) guidelines at a wavelength of 580 nm after derivatization with anisaldehyde reagent. Various bands were separated on TLC plate, highlighting the presence of different phytoconstituents. The interpretation of the m/z ratio of the mass spectra of a selected TLC band revealed the presence of cassane diterpene β-caesalpin and α-caesalpin at RF values of 0.64 and 0.78, respectively. For both caesalpins, a linear response was observed in the concentration range of 200–1200 ng/band. In C. bonduc, the TLC‒MS method was successfully applied to identify β-caesalpin and α-caesalpin. For the determination of β- and α-caesalpin in the hydroalcoholic leaf extract of C. bonduc, the developed approach was revealed to be robust, precise, accurate, and specific.