Quantitation and stability of piperacillin and tazobactam in plasma and ultrafiltrate from patients undergoing continuous venovenous hemofiltration by HPLC

Simple and reproducible HPLC methods for the determination of piperacillin and tazobactam have been developed and a complete stability study carried out. The method for piperacillin plasma samples consisted of protein precipitation with methanol using penicillin G as internal standard. No sample preparation was needed for ultrafiltrate samples. Tazobactam sample preparation involved protein precipitation with acetonitrile and the removal of lipids with dichloromethane. Piperacillin separation was performed on a µBondapack™ C 18 column (300 × 3.9, 10 µm) and tazobactam on a Novapack™ C 18 column (150 × 3.9, 4 µm) with UV detection set at 229 and 225 nm, respectively. The mobile phase consisted of phosphate buffer–acetonitrile, delivered at 1.5 mL[sol ]min. Calibration curves determination coefficients were ≥0.999 and response factors CV% < 5%. Intra‐ and inter‐assay precision and accuracy of the quality control and limit of quantification were satisfactory. Plasma and ultrafiltrate samples were stable at −20 and −80°C for 2 months and after three freeze–thaw cycles. In the chromatographic rack, tazobactam ultrafiltrate samples were stable for 24 h and plasma samples for 12 h, piperacillin ultrafiltrate samples for 8 h, but plasma samples for only 4 h. Storage of piperacillin samples at 4°C until analysis is recommended. Piperacillin was stable in the presence of tazobactam. Copyright © 2005 John Wiley & Sons, Ltd.