DEVELOPMENT AND VALIDATION OF A RP-HPLC METHOD FOR THE SIMULTANEOUS DETERMINATION OF QUERCETIN, ELLAGIC ACID AND RUTIN IN HYDROALCOHOLIC EXTRACT OF TRIPHALA CHURNA


Sultana Shaikh, Vandana Jain

Abstract


Objective: To develop a novel, accurate, precise and linear reverse phase high performance liquid chromatographic (RP-HPLC) method for simultaneous qualitative and quantitative estimation of quercetin, ellagic acid and rutin in an ayurvedic formulation and validate as per international conference on harmonization (ICH) guidelines.

Methods: In the present work, good chromatographic separation was achieved isocratically using a shim-pack HPLC C18 column (4.6 x 250 mm, 5μm) and a mobile phase consisting of 0.02 M potassium dihydrogen orthophosphate buffer (pH adjusted to 3 with orthophosphoric acid) and methanol in the ratio 55:45, at flow rate of 1 ml/min and column temperature maintained at 35 °C. The effluents obtained were monitored at 254 nm with a UV-visible detector.

Results: The retention time of quercetin, ellagic acid and rutin were found to be 7.52 min, 9.10 min and 12.47 min respectively. Linearity of quercetin, ellagic acid and rutin were found in the range of 8-12 ppm, 9-17 ppm and 7-11 ppm respectively. The correlation coefficient for quercetin, ellagic acid and rutin were 0.997, 0.999 and 0.999 respectively. The high recovery values (98 %-102 %) indicate a satisfactory accuracy. The low percent relative standard deviation (% RSD) values in the precision study reveal that the method is precise.

Conclusion: The developed method is novel, simple, precise, rapid, accurate and reproducible for simultaneous quantitative estimation of quercetin, ellagic acid and rutin in an ayurvedic formulation. Hence the developed method can be used for quantitative analysis and quality control of extracts and commercial samples of other plant species and formulation containing these three markers.


Keywords


Quercetin, Ellagic acid, Rutin, Ayurvedic formulation, RP-HPLC, Validation, ICH

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References


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About this article

Title

DEVELOPMENT AND VALIDATION OF A RP-HPLC METHOD FOR THE SIMULTANEOUS DETERMINATION OF QUERCETIN, ELLAGIC ACID AND RUTIN IN HYDROALCOHOLIC EXTRACT OF TRIPHALA CHURNA

Keywords

Quercetin, Ellagic acid, Rutin, Ayurvedic formulation, RP-HPLC, Validation, ICH

DOI

10.22159/ijap.2018v10i3.25860

Date

08-05-2018

Additional Links

Manuscript Submission

Journal

International Journal of Applied Pharmaceutics
Vol 10, Issue 3 (May-Jun), 2018 Page: 169-174

Online ISSN

0975-7058

Authors & Affiliations

Sultana Shaikh
M. PHARM Student, Department of Quality Assurance, Oriental College of Pharmacy, Sanpada - Navi Mumbai, Maharashtra 400705
India

Vandana Jain
Department of Quality Assurance, Oriental College of Pharmacy, Sanpada - Navi Mumbai, Maharashtra 400705
India


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