Determination of 3,4-Dihydroxy Methyl Benzoate in Rat Plasma by HPLC and Its Pharmacokinetics

A high-performance liquid chromatography method was established for determination of 3,4-dihydroxy methyl benzoate in rat plasma and for investigation of pharmacokinetics after intragastric administration of 3,4-dihydroxy methyl benzoate water solution. 3,4-Dihydroxy methyl benzoate and internal standard were separated on a Diamonsil(TM) C18 column (250 mm × 4.6 mm, 5 µm), using a mobile phase of methanol-acetonitrile-water-glacial acetic acid (6 : 3 : 100 : 0.25, v/v/v/v), and the detection wavelength was 254 nm. The calibration curve of 3,4-dihydroxy methyl benzoate was linear in the concentration range of 0.1313-25.25 µg mL(-1). The lower limit of quantification was 0.1313 µg mL(-1), and the relative standard deviations of intra- and inter-day were <11.7%. The main pharmacokinetic parameters were as follows: Cmax (12.1 ± 1.93) mg L(-1), Tmax (27.5 ± 6.12) h, t1/2 (39.2 ± 5.66) h, AUC(0→t)(0.85 ± 0.13) g min L(-1), AUC(0→∞) (0.88 ± 0.14) g min L(-1), MRT (70.7 ± 5.27) min(-1), Vd (0.32 ± 0.04) L and CL (5.79 ± 0.82) mL min(-1). The method is proven to be suitable for the pharmacokinetic study of 3,4-dihydroxy methyl benzoate, which offers the advantage of simplicity, sensitivity and accuracy.