Shivangi J Patel, Hardik Parmar, and Kirti V Patel*

Faculty of Pharmacy, The Maharaja Sayajirao University of Baroda, 390001, Vadodara, Gujarat, India

*Corresponding Author: Kirti V Patel, Faculty of Pharmacy, The Maharaja Sayajirao University of Baroda, 390001, Vadodara, Gujarat, India.

Received: September 11, 2025; Published: October 09, 2025

Abstract

Background: Kachnardi yoga a traditional medicine used to treat several bacterial infection and serious diseases, had also proven beneficial for the usage in breast cancer due to the presence of Kaempferol and Quercetin.

Purpose: The purpose of the study is to developed a bioanalytical technique for concurrently estimating KML and QRN from ethanolic extract of kachnardi yoga in mice plasma using RP-HPLC using Naringenin as internal standard.

Material and Method: The Gradient elution technique was developed to separate KML, QRN, and NGN within 20 minutes of total run time, where the mobile phase consisted of acetate buffer pH 5.35 and Acetonitrile in a Linear gradient pattern. The flow rate of 1.0 mL/min, and the injection volume of 50 µL. Detection was achieved at a wavelength of 375 nm, and the retention times for KML, QRN, and NGN were found to be 5.2, 7.2, and 10.2 minutes, respectively. Mice Plasma samples were processed using a protein precipitation technique to extract the analytes.

Results: The method demonstrated linearity with correlation coefficients (R²) of 0.9994 for KML and 0.9992 for QRN, over a concentration of 75-1200 ng/mL and 35-1000 ng/mL, respectively. The results of all the validation parameters performed as per ICH guidelines M10 were within the accepted limits with less than 3 %RSD. The % recovery from plasma was more than 95%. Freeze- thaw, Bench-top, short-term, and long-term stability studies were also achieved at LQC, MQC, and HQC levels.

Conclusion: The established bioanalytical technique can simultaneously effectually estimate the KML and QRN in kachnardi yoga from plasma samples and is suitable for further pharmacokinetic studies.

Keywords: Kaempferol; Quercetin; Simultaneous Estimation Bioanalytical Method; RP-HPLC; Mice Plasma; Stability Studies; Bioanalytical Method Validation

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Citation

Citation: Kirti V Patel., et al. “RP-HPLC Bioanalytical Simultaneous Estimation of Kaempferol and Quercetin in Mice Plasma". Acta Scientific Pharmaceutical Sciences 9.10 (2025): 57-69.

Copyright

Copyright: © 2025 Kirti V Patel., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.