DEVELOPMENT AND VALIDATION OF A HIGH PERFORMANCE LIQUID CHROMATOGRAPHY METHOD TO DETERMINE VANCOMYCIN CONCENTRATIONS IN PLASMA AND PIG PULMONARY TISSUE

Abstract

Simple and specific analytical methods were developed and validated to quantify vancomycin in plasma and lung tissue, which were obtained from a model of pneumonia in mechanically ventilated piglets. Tissue and plasma samples were precipitated and centrifuged; 100 µL of the supernatant were injected into the chromatographic system. Ceftazidime was used as the internal standard. The stationary phase was a silica based column Symmetry300 C₁₈ (150 × 4.6 mm) with pre-column. The mobile phase consisted of 20% ultrafiltered water and 80% of (A) 75 mM sodium acetate buffer (pH = 3) with (B) acetonitrile (92%/8%;v/v). Isocratic flow rate was set at 0.8 mL/min and 0.7 mL/min for plasma and tissue samples, respectively. UV absorbance detection was set at 230 nm. Standard curves showed good linearity for plasma and pulmonary tissue (r² > 0.99). The lower limits of quantitation were 1.56 µg/mL and 3.13 µg/mL for plasma and tissue, respectively. The intra- and inter-day precisions and accuracy all satisfied the acceptance criteria. Both HPLC assays to quantify vancomycin in plasma and pulmonary tissue are rapid, simple, and inexpensive. These methods could be helpful to develop further pharmacokinetic studies of vancomycin penetration in pulmonary tissue.

Keywords:

• animal model
• high performance liquid chromatography
• lung tissue
• pharmacokinetics
• plasma
• vancomycin

ACKNOWLEDGMENTS

This study was funded by grants from Ciber de Enfermedades Respiratorias (CibeRes, CB06/06/0028) from Instituto de Salud Carlos III (ISCIII), Fondo de Investigaciones Sanitarias (FIS) beca FIS PI070419 and 2009 SGR-911.

Notes

^a Vancomycin area/Ceftazidime area.

^b p < 0.05 to be significant.

^a Vancomycin area/Ceftazidime area.

^b p < 0.05 to be significant.