Development and Validation of High‑performance Thin‑layer Chromatography Method for Simultaneous Determination of Polyphenolic Compounds in Medicinal Plants
C. V. Jayachandran Nair1, Sayeed Ahamad2, Washim Khan2, Varisha Anjum2, Rajani Mathur1★★ Corresponding author
- 1Department of Pharmacology, Delhi Institute of Pharmaceutical Sciences and Research, INDIA.
- 2Department of Pharmacognosy and Phytochemistry, Bioactive Natural Product Laboratory, Faculty of Pharmacy, Jamia Hamdard, New Delhi, INDIA.
CORRESPONDENCE
Rajani Mathur
Department of Pharmacology, Delhi Institute of Pharmaceutical Sciences and Research, INDIA.
Volume 9, Issue 5S · pp. s67–s73 · PUBLISHED · DOI: 10.4103/pr.pr_122_16
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ABSTRACT
Context:Quantitative standardization of plant-based products is challenging albeit essential to maintain their quality. Aims:This study aims to develop and validate high-performance thin-layer chromatography (HPTLC) method for the simultaneous determination of rutin (Ru), quercetin (Qu), and gallic acid (Ga) from Psidium guajava Linn. (PG) and Aegle marmelos (L.) Correa. (AM) and correlate with antioxidant activity. Materials and Methods:The stock solution (1 mg/mL) of standard Ru, Qu, and Ga in methanol: Water (1:1) was serially diluted and spotted (5 μL) on slica gel 60 F254thin-layer chromatography plates. Toluene: Ethyl acetate: Formic acid: Methanol (3:4:0.8:0.7, v/v/v) was selected as mobile phase for analysis at 254 nm. Hydroalcoholic (1:1) extracts of leaves of PG and AM were fractionated and similarly analyzed. Antioxidant activity was also determined using 2, 2-diphenyl-1-picrylhydrazyl assay. Results: The developed method was robust and resolved Ru, Qu, and Ga at Rf0.08 ± 0.02, 0.76 ± 0.01, and 0.63 ± 0.02, respectively. The intra-day, interday precision, and interanalyst were <2% relative standard deviation. The limit of detection and limit of quantification for Ru, Qu, and Ga were 4.51, 4.2, 5.27, and 13.67, 12.73, 15.98 ng/spot, respectively. Antioxidant activity (Log 50% inhibition) of PG and AM was 4.947 ± 0.322 and 6.498 ± 0.295, respectively. Conclusion: The developed HPTLC method was rapid, accurate, precise, reproducible, and specific for the simultaneous estimation of Ru, Qu, and Ga.
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Nair, C. V. J., Ahamad, S., Khan, W., Anjum, V., & Mathur, R. (). Development and Validation of High‑performance Thin‑layer Chromatography Method for Simultaneous Determination of Polyphenolic Compounds in Medicinal Plants. Pharmacognosy Research, 9(5S), s67–s73. https://doi.org/10.4103/pr.pr_122_16