[Determination of metronidazole in serum by HPLC]

This paper report a sensitive and rapid method for the determination of metronidazole (MTZ) using theophylline as the internal standard. High performance liquid chromatograph model 344 (Beckman) with a 254 nm wavelength UV detector and YWG-C18H37 column (10 microns, 250 x 4.6 mm) was used. To the serum sample 200 microliters, 100 microliters phosphate buffer (0.8 mol/L, pH 7.5) was added, then extracted with 3 ml chloroform containing 5% isopropyl alcohol. The organic layer was removed and evaporated to dryness under an air stream in a 40 degrees C water bath. The residue was dissolved in 30 microliters mobile phase and 20 microliters injected. The mobile phase of water-methanol (73:27) was pumped at 1.0 ml/min through the column. The detector operated at 0.005 aufs. The retention times for MTZ and theophylline were 5.78 and 6.81 min respectively. Standard curve was linear in the concentration range of 0.3125 to 20 mg/L. The detection limit in serum was 0.02 mg/L. Extraction recovery was 77%-82%; method recovery 99%-102%; withinday RSD less than 3.0%; inter-day RSD less than 3.5%.