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

Biomed Chromatogr. 2005 Oct;19(8):570-8. doi: 10.1002/bmc.482.


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 microBondapack C(18) column (300 x 3.9, 10 microm) and tazobactam on a Novapack C(18) column (150 x 3.9, 4 microm) 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 >or=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 degrees 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 degrees C until analysis is recommended. Piperacillin was stable in the presence of tazobactam.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Anti-Bacterial Agents / analysis*
  • Anti-Bacterial Agents / blood
  • Calibration
  • Chromatography, High Pressure Liquid / methods*
  • Hemofiltration / methods*
  • Humans
  • Penicillanic Acid / analogs & derivatives*
  • Penicillanic Acid / analysis
  • Penicillanic Acid / blood
  • Piperacillin / analysis*
  • Piperacillin / blood
  • Sensitivity and Specificity
  • Tazobactam
  • Ultrafiltration


  • Anti-Bacterial Agents
  • Penicillanic Acid
  • Tazobactam
  • Piperacillin