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Simultaneous identification of rutin, chlorogenic acid and gallic acid in Moringa oleifera by densitometric high-performance thin-layer chromatography method

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Abstract

An HPTLC (high-performance thin-layer chromatography) method was carried out to develop a feasible, simple and accurate method for the estimation of exact concentration of flavonoids in Moringa oleifera. The simultaneous determination of rutin, chlorogenic acid, and gallic acid extracted from different parts (leaf, stem, flower, and root) of M. oleifera was performed and analysis was carried out on glass-backed plates (10 cm × 20 cm) coated silica gel 60 F254 (0.2 mm thickness, Merck, Germany) under CAMAG TLC Scanner III with UV densitometrically at λmax = 254 nm. The mobile phase of ethyl acetate:acetone:water:formic acid with a ratio of 6:3:2:2 (v/v) was used for these flavonoids. The findings of HPTLC gave sharp, symmetrical and well-resolved peaks at Rf values of 0.51 ± 0.02, 0.72 ± 0.04 and 0.83 ± 0.03 with linearity ranging 100–700 ng/spot (r2 = 0.9989), (r2 = 0.9992), and (r2 = 0.9984) for rutin (0.066 ± 0.003%), chlorogenic acid (0.033 ± 0.005%), and gallic acid (0.031 ± 0.005%), respectively. The developed method can also be used as a platform for estimation of complex bioactive(s) bio-synthesized in medicinal or aromatic plants.

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Acknowledgments

This project was supported by the Deanship of Scientific Research at Prince Sattam bin Abdulaziz University under the research project #2019/01/10418. Logistical support was provided by the Sara bint Rached bin Ghonaim Research Chair for Cultivating Non-Traditional Medicinal and Aromatic Plants, Biology Department, College of Science and Humanities, Prince Sattam bin Abdulaziz University, Alkharj, Saudi Arabia.

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Author notes

  1. Pravej Alam and Prawez Alam contributed equally to this work.

Authors and Affiliations

  1. Department of Biology, Sara Alghonaim Research Chair (SRC), College of Science and Humanities, Prince Sattam bin Abdulaziz University (PSAU), Alkharj, 11942, Saudi Arabia

    Pravej Alam & Mahmoud A. Sharaf-Eldin

  2. Department of Pharmacognosy, College of Pharmacy, Prince Sattam bin Abdulaziz University, P.O. Box 173, Alkharj, 11942, Saudi Arabia

    Prawez Alam & Mohammed H. Alqarni

  3. Medicinal and Aromatic Plants Research Department, Non Traditional Spices Biotechnology Unit, National Research Centre, Cairo, 12622, Egypt

    Mahmoud A. Sharaf-Eldin

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  1. Pravej Alam
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  2. Prawez Alam
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  3. Mahmoud A. Sharaf-Eldin
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  4. Mohammed H. Alqarni
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Contributions

PA and PA designed the work and performed the experiment while MAS and MA evaluated the data and presented in this article.

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Correspondence to Pravej Alam.

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Alam, P., Alam, P., Sharaf-Eldin, M.A. et al. Simultaneous identification of rutin, chlorogenic acid and gallic acid in Moringa oleifera by densitometric high-performance thin-layer chromatography method. JPC-J Planar Chromat 33, 27–32 (2020). https://doi.org/10.1007/s00764-019-00002-2

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  • DOI: https://doi.org/10.1007/s00764-019-00002-2

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