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<p>This study outlines the development and validation of a High-Performance Thin-Layer Chromatography (HPTLC) method for the precise estimation of Cuminaldehyde and β-Sitosterol in AYUSH SS Granules, a complex herbal formulation. The optimal mobile phases, Toluene: Ethyl Acetate: Acetic Acid (9:0.8:0.2 v/v/v) for Cuminaldehyde and Benzene: Ethyl Acetate: Formic Acid (6:4.5:0.5 v/v/v) for β-Sitosterol, achieved clear separations and distinct peak profiles. Method validation demonstrated strong linearity for both markers, with correlation coefficients (r<sup>2</sup>) of 0.9918 for Cuminaldehyde and 0.9994 for β-Sitosterol. Specificity was confirmed by consistent R<sub>f</sub> values and peak purity across standards and samples, as supported by 2D-densitograms. The method displayed excellent precision with intra-day and inter-day RSD values below 2%, indicating reproducibility. Accuracy was affirmed by recovery studies yielding 98–102% recovery rates for both markers. Despite procedural variations, robustness and ruggedness were validated with RSDs under 2%, supporting the method’s stability across different conditions and analysts. Sensitivity was notable, with low LOD and LOQ values of 29.66&#xa0;ng and 89.89&#xa0;ng for Cuminaldehyde and 2.22&#xa0;ng and 6.72&#xa0;ng for β-Sitosterol, respectively, underscoring its capability for detecting trace bioactive compounds. Application to AYUSH SS Granules confirmed formulation consistency, with quantified concentrations of 0.2223 ± 0.0019&#xa0;mg/g for Cuminaldehyde and 0.1619 ± 0.0005&#xa0;mg/g for β-Sitosterol. The developed HPTLC method proves to be a reliable, precise, and sensitive tool, well-suited for routine quality control of herbal formulations, to know the presence of Cuminaldehyde, β-Sitosterol. Further, ensuring compliance with bioactive marker specifications is critical for therapeutic efficacy and safety.</p>

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Validated HPTLC method for quantification of Cuminaldehyde and β-Sitosterol in AYUSH SS granules

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年9月14日

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  • Lakshminarayana Misro,
  • Thirupataiah Boini,
  • Rahul Maurya,
  • R. Thulasi,
  • Vijay Kumar,
  • Ajay Kumar Meena,
  • Ravindra Singh,
  • Arjun Singh,
  • N. Srikanth,
  • Rabinarayan Acharya

摘要

This study outlines the development and validation of a High-Performance Thin-Layer Chromatography (HPTLC) method for the precise estimation of Cuminaldehyde and β-Sitosterol in AYUSH SS Granules, a complex herbal formulation. The optimal mobile phases, Toluene: Ethyl Acetate: Acetic Acid (9:0.8:0.2 v/v/v) for Cuminaldehyde and Benzene: Ethyl Acetate: Formic Acid (6:4.5:0.5 v/v/v) for β-Sitosterol, achieved clear separations and distinct peak profiles. Method validation demonstrated strong linearity for both markers, with correlation coefficients (r2) of 0.9918 for Cuminaldehyde and 0.9994 for β-Sitosterol. Specificity was confirmed by consistent Rf values and peak purity across standards and samples, as supported by 2D-densitograms. The method displayed excellent precision with intra-day and inter-day RSD values below 2%, indicating reproducibility. Accuracy was affirmed by recovery studies yielding 98–102% recovery rates for both markers. Despite procedural variations, robustness and ruggedness were validated with RSDs under 2%, supporting the method’s stability across different conditions and analysts. Sensitivity was notable, with low LOD and LOQ values of 29.66 ng and 89.89 ng for Cuminaldehyde and 2.22 ng and 6.72 ng for β-Sitosterol, respectively, underscoring its capability for detecting trace bioactive compounds. Application to AYUSH SS Granules confirmed formulation consistency, with quantified concentrations of 0.2223 ± 0.0019 mg/g for Cuminaldehyde and 0.1619 ± 0.0005 mg/g for β-Sitosterol. The developed HPTLC method proves to be a reliable, precise, and sensitive tool, well-suited for routine quality control of herbal formulations, to know the presence of Cuminaldehyde, β-Sitosterol. Further, ensuring compliance with bioactive marker specifications is critical for therapeutic efficacy and safety.