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

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 mu m) 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 mu L/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.