A rapid and simple method of high performance liquid chromatography with UV detection for the quantification of vancomycin in artificial perfusion fluid and lung tissue samples has been developed and validated. Chromatographic separation was carried out in a Nucleosil 120 C(18) 5 microm column (length, 15 cm; inner diameter, 0.4 cm) using a mixture of 0.05 M NH(4)H(2)PO(4) (pH 4)-acetonitrile (92:8, v/v) as the mobile phase at a flow rate of 1 mL/min, with UV detection at 220 nm. The method used for the vancomycin quantification showed linearity for concentration ranges of 0.1-2, 2-15 and 15-250 microg/mL, with r(2)=0.9985, 0.9996 and 0.9985, respectively. The limit of quantification of the method was 0.1 microg/mL and the coefficients of variation of the between- and within-day precision showed values between 0.6% and 7.0%. The retention time of vancomycin was 8.5 min. The method was used successfully to study the pharmacokinetics of vancomycin in isolated rat lung after its administration through the systemic and inhalatory routes.