Determination of quercetin in human plasma using reversed-phase high-performance liquid chromatography |
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Affiliation: | 1. Department of Pharmaceutics, College of Pharmacy, Najran University, Saudi Arabia;2. Nanomedicine Research Lab, Department of Pharmaceutics, Faculty of Pharmacy, Jamia Hamdard, New Delhi 110062, India;;3. Irma Lerma Rangel College of Pharmacy, Texas A&M Health Science Center, Kingsville, Texas, USA |
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Abstract: | A method is reported for the measurement of quercetin in human plasma using reversed-phase high-performance liquid chromatography (HPLC). Quercetin and kaempferol (as internal standard) were spiked into plasma samples and extracted using C18 Sep-Pak Light cartridges (efficiency > 85%). Flavonoids were eluted with aqueous acetone (50% v/v, pH 3.5), dried down and redissolved in aqueous acetone (45% v/v, pH 3.5). The increased osmolarity promoted a phase separation and the water-saturated acetone layer, containing the flavonoids, was analysed by HPLC with aqueous acetone mobile phase (45% v/v acetone in 250 mM sodium dihydrogen sulphate. The mixture was adjusted to pH 3.5 with phosphoric acid and used at a flow-rate of 1.0 ml/min) and μBondapak C18 column (150 × 3.9 mm I.D., 10 μm particle size). The detection limit (A375 nm) for quercetin in plasma was 0.1 μg/ml (300 nM). The method also detects metabolites of quercetin, although these are not yet identified. |
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