Peer Reviewed Journal
2025, Vol. 5, Issue 2, Part A
Development and validation of a simple, accurate, and precise RP-HPLC method for the simultaneous estimation of hesperidin and quercetin in plant extracts
Author(s): Gaurav Yadav, Ravi Kumar, Muraree Lal and Avinash K Kondalkar
Abstract: A new RP-HPLC method was developed for the simultaneous estimation of hesperidin and quercetin in plant extracts and it was validated as per ICH Q2(R1) guidelines. The chromatographic separation was found to be satisfactory on a Symmetry C-18 column (4.6×150mm, 5μm Thermosil) using phosphate buffer (pH 4.5) and acetonitrile in the ratio of 30:70 v/v at a flow rate of 1.0 mL/min. The retention times of hesperidin and quercetin were found to be 6.94 min and 10.50 min respectively, with UV detection at 238 nm. The system suitability parameters confirmed that the proposed method is suitable for simultaneous estimation of hesperidin and quercetin with excellent chromatographic performance. The tailing factor for the peaks was found to be 0.995 and 1.203 for hesperidin and quercetin respectively, while the theoretical plates for separation were 3,605 and 7,581 respectively. The method demonstrated excellent linearity in the concentration range of 10-50 μg/mL for both compounds with correlation coefficients (r²) of 0.999. The precision of the method was exceptional with relative standard deviation values well within acceptable limits, and the recovery of both analytes was within the acceptance range of 98-102%. The limit of detection (LOD) was determined to be 0.003 μg/mL for hesperidin and 0.09 μg/mL for quercetin, while the limit of quantification (LOQ) was 0.012 μg/mL for hesperidin and 0.3 μg/mL for quercetin. The proposed RP-HPLC method was found to be suitable for the simultaneous estimation of hesperidin and quercetin in plant extracts and pharmaceutical formulations. The method is simple, selective, reproducible, and accurate with excellent precision, making it highly applicable for routine analytical purposes in quality control laboratories and pharmaceutical research institutions.
Pages: 37-43 | Views: 157 | Downloads: 52
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How to cite this article:
Gaurav Yadav, Ravi Kumar, Muraree Lal, Avinash K Kondalkar. Development and validation of a simple, accurate, and precise RP-HPLC method for the simultaneous estimation of hesperidin and quercetin in plant extracts. Nat J Pharm Sci 2025;5(2):37-43.
