JOURNAL OF PHARMACOLOGY AND
BIOMEDICINE
ISSN.2456-8244
A new RP-HPLC method was developed for the simultaneous estimation of hepseridine and quercetin was validated as per ICH guidelines. The chromatogram for was found to be satisfactory on symmetry C-18 (4.6×150mm, 5µ Hypersil column) using mobile phase composed of Methanol-Water (70:30, pH adjusted to 3.0 with orthophosphoric acid) at a flow rate of 1.0 ml/min and the detection wavelength of 269.7 nm. The retention time of hesperidine was found to be 2.547 min and that of quercetin was found to be 2.106 min. The system suitability parameters proved that the proposed method is suitable for estimation of both the drugs under study. The linearity for hesperidine was studied from 1 to 5 µg/ml concentrations and for quercetin was studied from 1-5µg/ml concentrations. The precision of the method was good and the recovery of drugs was found to be within the acceptance limits of 80-120%. The LOD and LOQ for hesperidine were found to be 0.084 µg/ml and 0.256 µg/ml respectively. The LOD and LOQ for quercetin were found to be 0.054 µg/ml and 0.166 µg/ml respectively. The proposed RP HPLC method was found suitable for the simultaneous estimation of hepseridine and quercetin and is simple, selective, reproducible and accurate with good precision and can be successfully applied to routine analytical purpose.
NAMES:
ONLINE ISSN:2456-8244
Keywords: Hesperidine, quercetin, HPLC, validation, simultaneous estimation
DOI:
1. https://go.drugbank.com/drugs/DB04703
2. https://pubchem.ncbi.nlm.nih.gov/compound/Hesperidin
3. https://pubchem.ncbi.nlm.nih.gov/compound/Quercetin
4. https://go.drugbank.com/drugs/DB04216
5. Sonawane SN, Kurangi BK, Shelar OA, Jalalpure AA, Gawas N, Koli R. Quality by design
assisted a stability indicating RP-HPLC method for the estimation of hesperidin in
transfersome and marketed product. Journal of the Indian Chemical Society. 2025; 102:
101521.
6. Chimagave SS, Jalalpure SS, Patil AK, Kurangi BK. Development and validation of
stability indicating RP-HPLC method for estimation of hesperidin in nanotransferosome
and Madhiphala rasayana—An Ayurvedic marketed product. Journal of Applied
Pharmaceutical Science. 2023; 13(2): 39-48
7. Carvalho D, Jesus Â, Pinho C, Oliveira RF, Moreira F, Oliveira AI. Validation of an
HPLC-DAD Method for Quercetin Quantification in Nanoparticles. Pharmaceuticals
2023, 16, 1736
8. Khursheed R, Singh SK, Kapoor B, Gulati M, Wadhwa S, Gupta S, Prasher P, Kumar D,
Dua K, Kumar LMS, Babu UV, Sharma M, Soni HK, Kumar V. Development and
Validation of RP-HPLC Method for Simultaneous Determination of Curcumin and
Quercetin in Extracts, Marketed Formulations, and Self-Nanoemulsifying Drug Delivery
System. Re:GEN Open. 2021; 1(1): 43-52
9. Alam P, Parvez MK, Arbab AH, Al-Dosari MS. Quantitative analysis of rutin, quercetin,
naringenin, and gallic acid by validated RP- and NP-HPTLC methods for quality control
of anti-HBV active extract of Guiera senegalensis. Pharmaceutical Biology. 2017; 55(1):
1317-1323
10. Adham AN. Qualitative and Quantitative Estimation of Hesperidin in Peel and Juice of
Citrus Fruits by RP-HPLC Method Growing in Kurdistan Region/Iraq. International
Journal of Pharmaceutical Sciences Reviews and Research. 2015; 33(2): 220-224
11. Biswas P, Jain P, Parkhe G, Mishra B. Validated HPLC method for the estimation of
Aspirin and Omeprazole in dosage form. J Pharmacol Biomed 2018; 2(3): 171-180.
12. Patel A, Soni P, Yadav N, Omray LK. New cost-effective RP-HPLC method development
and evaluation for quantitative estimation of Pitavastatin in pharmaceutical formulation. J
Pharmacol Biomed 2018; 2(3): 181-188