Determination of mirosamicin in animal tissues by high-performance liquid chromatography

A simple and rapid method using high-performance liquid chromatography (HPLC) for the determination of mirosamicin in animal tissues has been developed. The drug was extracted with 0.3% metaphosphoric acid-methanol (7:3, v/v), and the extracts were cleaned on a Bond Elut SCX (500 mg) cartridge. The HPLC separation was performed on a Puresil 5C18 column (150 x 4.6 mm I.D.) with 0.05 M phosphate buffer (pH 2.5)-acetonitrile (70:30) as the mobile phase at a flow-rate of 0.5 ml/min; the drug was detected at 230 nm with 0.04 AUFS. The calibration graph was linear from 5 to 100 ng. The recoveries of microsamicin from various animal tissues fortified at 1.0 microgram/g were 83.7-88.6% with a relative standard deviation (R.S.D.) of 2.0-5.7%. The detection limit was 0.05 microgram/g.