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Simultaneous determination of lupeol and β-sitosterol by high-performance thin-layer chromatographic method in Crataeva nurvala Buch-Ham. stem bark

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Abstract

Crataeva nurvala Buch-Ham. (Capparidaceae) is an important medicinal plant, the bark of which is extensively used in Ayurveda as a remedy for kidney stones, urinary tract infections, prostate-related disorders, and benign prostatic hypertrophy. In this study, a sensitive and reliable high-performance thin-layer chromatography (HPTLC) method has been developed for the simultaneous quantification of two bioactive markers, lupeol and β-sitosterol, in the stem bark of C. nurvala using precoated silica gel G 60 F254 HPTLC plates as the stationary phase and toluene–methanol (9.6:0.4, v/v) as the mobile phase. Detection and quantification were carried out by densitometric scanning at 545 nm after derivatization using 1% anisaldehyde–sulfuric acid reagent. The method was validated for linearity, inter-day and intra-day precision, repeatability, accuracy, specificity, limit of detection, and limit of quantification. The linear regression analysis data for the calibration plots showed a good linear relationship (r2 = 0.987–0.990) in the concentration range of 2 to 6 μg for lupeol and β-sitosterol, both with respect to areas. The proposed method provides good resolution of lupeol and β-sitosterol from other constituents present in the stem bark of C. nurvala. It was found to be simple, accurate, precise, and reproducible and would potentially be a useful tool for routine analysis of C. nurvala.

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Acknowledgments

The authors are thankful to L. M. College of Pharmacy, Ahmedabad, Gujarat, for providing the necessary facilities to conduct this research work and Dr. Hitesh A. Solanki, Professor and Taxonomist, Department of Botany, Gujarat University, Ahmedabad, Gujarat, for the authentication of the plant material used in the study.

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Funding

The research was supported by the L. M. College of Pharmacy, Navrangpura, Ahmedabad, Gujarat, India.

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Authors and Affiliations

  1. Department of Pharmacognosy, L. M. College of Pharmacy, Navrangpura, Ahmedabad, Gujarat, 380009, India

    Pradeep A. Sadhwani, Preeti D. Verma & Mamta B. Shah

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  1. Pradeep A. Sadhwani
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  2. Preeti D. Verma
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  3. Mamta B. Shah
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Contributions

Pradeep A. Sadhwani (M. Pharm. student) contributed in performing the laboratory work and chromatographic analysis and drafted the paper. Dr. Preeti D. Verma co-supervised the work and contributed to analysis of the data as well as reviewed the manuscript. Dr. Mamta B. Shah was responsible for the project concept, design, and supervision of the entire study and contributed to critical reading of the manuscript. All the authors have read the final manuscript and approved the submission.

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Correspondence to Mamta B. Shah.

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Sadhwani, P.A., Verma, P.D. & Shah, M.B. Simultaneous determination of lupeol and β-sitosterol by high-performance thin-layer chromatographic method in Crataeva nurvala Buch-Ham. stem bark. JPC-J Planar Chromat 33, 365–370 (2020). https://doi.org/10.1007/s00764-020-00043-y

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