Abstract
A precise, swift and environmental-friendly reverse phase ultra-high performance liquid chromatographic assay for the determination of thymoquinone (TQ) in plasma samples using thymol (TM) as an internal standard was developed and validated. The method used a high strength silica C18 1.7μm column (100×2.1mm) with an isocratic mobile phase consisting of a blend of methanol and 20mM potassium dihydrogen ortho-phosphate (90:10v/v; pH of 4.2). The selected eluent provided a short run time (≤2min), better peak symmetry, lower limit of quantification of 10ng/mL and satisfactory values of other chromatographic parameters including resolution (Rs=1), capacity factor (k=21.5 and 14.5 for TQ and TM, respectively), selectivity (α=1.482) and number of theoretical plates (N=1653 and 784 for TQ and TM, respectively). The method was efficiently applied to a pharmacokinetic study of TQ following an intraperitoneal administration of 2mg/kg in mice. The concentrations of TQ in plasma were measurable up to 12h with Cmax of 404.08±28.91ng/mL, T1/2 of 2.31±0.10h and area under plasma concentration-time curve of 1527.00±46.61ng/mL×h.
Published Version
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