Method Development and Validation for Quantification of Quercetin in Blood Plasma of Wistar Rats Using LC-MS/MS

Abstract

Background: Quercetin is a flavanol that has demonstrated pharmaceutical properties such as anti-inflammatory, antioxidant, and anti-carcinogenic properties. However, parenteral formulations of quercetin are currently not in widespread use due to its poor aqueous solubility, fast metabolism, and low bioavailability. Objective: This study aimed to develop a quick, simple, and accurate method for the quantification of quercetin using ultra-fast liquid chromatography-tandem mass spectrometry (LC-MS/MS). Methods: A rapid and sensitive LC-MS/MS method was developed and fully validated for the quantification of quercetin in rat plasma after intravenous administration. Quercetin reference standard was used for the method development as well as in-vivo validation. ACQUITY UPLC BEH C18 Column (2.1 x 50 mm, 1.7 μm) was used for the chromatographic separation. The mobile phase consisted of solution A (water with 0.1% formic acid) and solution B (methanol: acetonitrile with 0.1% formic acid in the ratio of 1:1) with a flow rate of 0.350 μL/min. Further, the method was applied to quantify and pharmacokineticly profile quercetin in the blood plasma of outbred male and female Wistar rats. Results: The accuracy of the method was calculated for intraday and inter-day, which came out as ≤ 85.23% and 84.87%, respectively. The precision of the method calculated for intraday was ≤3.02%, and for inter-day was ≤2.75%. The percentage recovery was found to be ≤85.23% for intraday and ≤84.87% for inter-day. The Relative Standard Deviation was 0.04 for intraday and 1.10 for inter day, respectively. Conclusion: The developed method has an advantage over other reported methods because of its short run time of 6 minutes as well as its short time of data registration of approximately 2.5 minutes. This method can be used for the routine analysis of quercetin in vivo animal studies.