Sensitive semi-microcolumn high-performance liquid chromatographic method for the determination of DU-6681, the active parent drug of a new oral carbapenem antibiotic, DZ-2640, in human plasma and urine using a column-switching system as sample clean-up procedure

DZ-2640 is a new oral carbapenem antibiotic having a dihydro-pyrroloimidazole ring as a side chain and a pivaloyloxymethyl (POM) ester prodrug of DU-6681, the active parent compound. A simple and sensitive column-switching semi-microcolumn high-performance liquid chromatographic method for the determination of DU-6681 in human plasma and urine has been developed. Human plasma was diluted with an equal volume of 1 M MOPS buffer (pH 7.0) and the mixture was filtered through an Ultrafree C3GV. The resulting filtrate was injected without further cleanup onto the HPLC system. Human urine was diluted with an equal volume of 1 M MOPS buffer (pH 7.0) and the mixture was directly injected onto the HPLC system. The analyte was detected by monitoring the column effluent with UV light at a wavelength of 300 nm, which resulted in the Limit of quantitation of 0.008 mu g/ml of plasma and 0.32 mu g/ml of urine. Calibration curves were Linear in the range of 0.008 to 5.85 mu g/ml in plasma and 0.32 to 104.4 mu g/ml in urine. The present methods showed greatly increased sensitivity for DU-6681 compared to conventional HPLC methods and also showed satisfactory recovery, selectivity, precision, and accuracy. Stability studies showed that 1 M MOPS buffer (pH 7.0) acted as a stabilizer. In plasma and urine diluted with equal volume of the buffer, DU-6681 showed good stability at -80 degrees C for up to 4 weeks with no significant loss of the drug. (C) 1999 Elsevier Science B.V. All rights reserved.