Rapid and Sensitive LC-MS/MS Method for the Determination of Metoprolol in Beagle Dog Plasma with a Simple Protein Precipitation Treatment and Its Pharmacokinetic Applications

A rapid LC-MS/MS method with good accuracy and sensitivity was developed and validated for the pharmacokinetics study of metoprolol (MP) in beagle dogs. The plasma samples were simply precipitated by methanol and then analyzed by LC-MS/MS. An Ultimate XB-C-18 column (150 x 2.1 mm ID, 5 mu m) was used for separation, with methanol-water containing 0.2% formic acid (65: 35, v/v) as the mobile phase at a flow rate of 0.2 mL/min. Monitoring ions of MP and internal standard (hydroxypioglitazone) were m/z 268.1/115.6 and m/z 373.1/150.2, respectively. The linear range was 3.03-416.35 ng/mL with an average correlation coefficient of 0.9996, and the limit of quantification was 3.03 ng/mL. The intra- and inter-day precision was less than 15%. At low, middle and high concentrations, the recovery, the matrix effect and the accuracy was in the range of 76.06%-95.25%, 93.67%-104.19% and 95.20%-99.96% respectively. The method was applied for the pharmacokinetics study of MP tartrate tablets (50 mg). The AUC(0-t), T-max and C-max were respectively 919.88 +/- 195.67 mu g/L.h, 0.96 +/- 0.33 h, 349.12 +/- 78.04 ng/mL.