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Bioanalytical Method Development and Validation for Herbal Quercetin in Nano Formulation by RP-UFLC in Rabbit Plasma

Shanmugam R, Gowthamarajan K, Priyanka DL, Madhuri K and Narayanareddy Karri VVS

A sensitive reverse phase ultra fast liquid chromatography (RP-UFLC) has been developed and validated for the quantification of Quercetin in nano formulation in rabbit plasma using solid phase extraction method (SPE). Best chromatographic resolution was achieved on a reverse phase Hibar®C18 (250×4.6 mm i.d., 5 μ) column with the mobile phase consisted of acetonitrile: pottasium dihydrogen ortho phosphate (pH 3.5) in the ratio of 65:35 (v/v) with a flow rate of 0.8 mL/min. The retention time for the drugs was found to be 8.6 min and 10.0 min for internal standard (fluticasone propionate). The analyte was detected by using a Photo-diode array (PDA) detector with LC solution software. Linearity was obtained in the concentration range of 10 to 400.0 ng/mL (r2= 0.989). Lower limit of quantification (LLOQ) was found to be 10 ng/mL. Middle limit of quantification (MQC) was found to be 200 ng/mL and higher limit of quantification (HQC) was found to be 380 ng/mL. The average recovery of the analyte was found to ranging from 95.91 to 98.59%. The developed method isapplicablefor estimation of Quercetin in nano-formulation, routine quality control analysis, pharmacokinetic and bioequivalence studies.