OBJECTIVE  To establish an UHPLC-MS/MS method to determine the concentration of ten constituents(ferulic acid, kaempferol, oleanolic acid, ligustilide, quercetin, astragaloside, betaine, calycosin, calycosin glucoside, poriacolic acid) in rat plasma, and to study the pharmacokinetic characteristics of them in rat in vivo.

METHODS  The plasma sample was treated with a liquid-liquid extraction of ethyl acetate, and glibenclamide was used as an internal standard. The determination was carried out with a Thermo Hypersil Gold column(2.1 mm×100 mm, 1.9 μm) and the mobile phase comprised of water containing 0.1% formic acid(A) and methanol(B). The flow rate, column temperature and injection volume were 0.3 mL·min^–1, 40 ℃ and 2 μL. The electrospray ionization source(ESI) and parallel reaction detection mode were used for quantitative analysis. After oral administration, the concentrations of 10 constituents in rat plasma were measured and pharmacokinetic parameters were calculated using DAS 2.0 software.

RESULTS  The linear relationship of 10 components in plasma was good(r>0.991), and the RSD of intra-day and inter-day precision of each component was <14%. The extraction recoveries of low, medium and high concentrations of each component in plasma were 79.41%−93.63%, and the matrix effect was 83.29%−106.29%, which met the requirements of biological sample determination. After rats were intragastrically administrated with Liuwei Tiaogeng tablets, the blood concentration peak time(T_max) of the 10 constituents was 0.25–4.96 h, the area under the blood concentration-time curve(AUC_0–∞) was 12.19–3 488.29 ng·mL⁻¹·h, and the blood concentration peak concentration(C_max) was 0.81–143.33 ng·mL⁻¹.

CONCLUSION  The method is specific and repeatable, and suitable for determination of ten constituents of Liuwei Tiaogeng tablets in rat plasma for pharmacokinetic study.