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<p>Phenolics, withanolides, and triterpenoids are prominent bioactive classes found in various adaptogenic botanicals and have potential applications as phytogenic feed additives. In the present study, a robust and reproducible instrumental thin-layer chromatography (TLC) method, coupled with densitometric analysis, was developed and validated for the simultaneous detection and quantification of these phytoconstituents in selected medicinal plant extracts. Method optimization involved selecting suitable mobile phases and derivatization reagents to achieve efficient resolution and reliable detection. Validation was carried out in accordance with the International Council for Harmonisation (ICH) guidelines, assessing parameters including specificity, linearity, precision, accuracy, limit of detection, and limit of quantification. The developed instrumental TLC method was successfully applied to standardize multiple adaptogenic botanical extracts, confirming the presence and concentration range of the targeted compounds. This approach facilitates quality control and ensures consistency in formulations intended for use as phytogenic feed additives. The study underscores the relevance of instrumental TLC as a cost-effective and high-throughput analytical tool for the phytochemical standardization of complex herbal matrices.</p> Graphical abstract <p></p>

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Development and validation of a novel method of analysis for phenolics, withanolides, and triterpenoids by instrumental thin-layer chromatography from adaptogenic botanicals in phytogenic feed additive

  • 期刊论文

  • 发表日期：

    2026年2月24日

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  • Swapnil Kamble,
  • Aboli Girme,
  • Ashish Chinchansure,
  • M. Deepak

摘要

Phenolics, withanolides, and triterpenoids are prominent bioactive classes found in various adaptogenic botanicals and have potential applications as phytogenic feed additives. In the present study, a robust and reproducible instrumental thin-layer chromatography (TLC) method, coupled with densitometric analysis, was developed and validated for the simultaneous detection and quantification of these phytoconstituents in selected medicinal plant extracts. Method optimization involved selecting suitable mobile phases and derivatization reagents to achieve efficient resolution and reliable detection. Validation was carried out in accordance with the International Council for Harmonisation (ICH) guidelines, assessing parameters including specificity, linearity, precision, accuracy, limit of detection, and limit of quantification. The developed instrumental TLC method was successfully applied to standardize multiple adaptogenic botanical extracts, confirming the presence and concentration range of the targeted compounds. This approach facilitates quality control and ensures consistency in formulations intended for use as phytogenic feed additives. The study underscores the relevance of instrumental TLC as a cost-effective and high-throughput analytical tool for the phytochemical standardization of complex herbal matrices.

Graphical abstract