Validation of a new HPLC-UV method for determination of the antibiotic linezolid in human plasma and in bronchoalveolar lavage

Abstract

A rapid and selective HPLC-UV method was developed for the quantification of linezolid (LNZ) in human plasma and bronchoalveolar lavage (BAL) at the concentrations associated with therapy. Plasma samples were extracted by solid-phase extraction followed by evaporation to dryness and reconstitution in mobile phase solution. The chromatographic separation was carried out on a C18 column with an isocratic mobile phase consisting of dihydrogen phosphate buffer 50 mm (pH 3.5) and acetonitrile (60:40 v/v). The detection was performed using a photodiode array. Under these conditions, a single chromatographic run could be completed within 12 min. The method was validated by estimating the precision and the accuracy for inter- and intra-day analysis in the concentration range of 25-25600 ng/mL. The method was linear over the investigated range with all the correlation coefficients R > 0.999. The intra- and inter-day precision was within 8.90% and the accuracy ranged from -4.76 to +5.20%. This rapid and sensitive method was fully validated and could be applied to pharmacokinetic study for the determination of LNZ levels in human plasma and BAL samples.

Keywords: BAL; HPLC-UV; linezolid; solid-phase extraction; therapeutic drug monitoring.