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Comparative Thin-Layer Chromatographic Studies and Development of a High-Performance Thin-Layer Chromatography Method for the Quantification of Lawsone in Natural and Micropropagated Plant Parts of Lawsonia inermis L.

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Summary

A simple, sensitive, and precised high-performance thin-layer chromatography (HPTLC) method has been developed for the analysis of lawsone in natural vis-à-vis micropropagated plant parts of Lawsonia inermis L. Separation of the components was perfectly achieved on high-performance thin-layer chromatography (TLC) plates using optimized tertiary mobile phase of benzene–ethyl acetate– acetic acid (7.5:2.5:0.1, v/v). Densitometric scanning was performed before derivatization of the plate in absorption/reflection mode, and lawsone was quantified at its maximum absorbance of wavelength of 275 nm. Linearity of the method was obtained in the concentration range of 50 to 350 ng spot−1 with a correlation coefficient (r2) of 0.9999, indicating good relationship between concentrations in opposition to the peak area. The limit of detection and limit of quantification were found to be 16 and 50 ng spot−1, respectively. The obtained recovery ranges from 95.09% to 96.90% with an average value of 96.02% proved the excellent accuracy of the method. The developed method was found to be highly sensitive, and the mobile phase enables outstanding separation of lawsone from other components present in the mixture. The International Conference on Harmonization (ICH) guidelines were followed for validation of the HPTLC method in terms of precision, repeatability, and accuracy. The maximum content of lawsone was reported in the leaves of the micropropagated plant.

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Authors and Affiliations

  1. Department of Botany and Biotechnology, Ravenshaw University, 753 003, Cuttack, Odisha, India

    Arpita Moharana, Durga Prasad Barik & Soumendra Kumar Naik

  2. Orissa University of Agriculture and Technology, 751003, Bhubaneswar, Odisha, India

    Kedar Kumar Rout

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  1. Arpita Moharana
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  2. Durga Prasad Barik
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  3. Soumendra Kumar Naik
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Correspondence to Kedar Kumar Rout.

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Moharana, A., Barik, D.P., Naik, S.K. et al. Comparative Thin-Layer Chromatographic Studies and Development of a High-Performance Thin-Layer Chromatography Method for the Quantification of Lawsone in Natural and Micropropagated Plant Parts of Lawsonia inermis L.. JPC-J Planar Chromat 31, 155–162 (2018). https://doi.org/10.1556/1006.2018.31.2.10

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