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 1.7 μm column (100 × 2.1 mm) with an isocratic mobile phase consisting of a blend of methanol and 20 mM potassium dihydrogen ortho-phosphate (90:10 v/v; pH of 4.2). The selected eluent provided a short run time (≤2 min), better peak symmetry, lower limit of quantification of 10 ng/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 2 mg/kg in mice. The concentrations of TQ in plasma were measurable up to 12 h with C_max of 404.08 ± 28.91 ng/mL, T_1/2 of 2.31 ± 0.10 h and area under plasma concentration-time curve of 1527.00 ± 46.61 ng/mL × h.

中文翻译：

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快速，环保的超高效液相色谱测定方法，用于测定血浆样品中的胸腺醌及其分析应用。

开发并验证了以百里酚（TM）为内标测定血浆样品中胸腺醌（TQ）的精确，快速，环境友好的反相超高效液相色谱法。该方法使用高强度硅胶C ₁₈ 1.7μm色谱柱（100×2.1 mm），其等度流动相由甲醇和20 mM正磷酸二氢钾（90:10 v / v； pH值为4.2）的混合物组成 。选择的洗脱液运行时间短（≤2 min），峰对称性更好，定量限较低（10 ng / mL）和其他色谱参数令人满意的值，包括分辨率（Rs = 1），容量系数（k  = 21.5和14.5）分别用于TQ和TM），选择性（α = 1.482）和理论塔板数（ TQ和TM分别为N = 1653和784）。该方法在小鼠腹膜内给药后有效地应用于TQ的药代动力学研究。血浆中TQ浓度可测量长达12 h，C _max为404.08±28.91 ng / mL，T _1/2为2.31±0.10 h，在血浆浓度-时间曲线下的面积为1527.00±46.61 ng / mL×h。