High performance liquid chromatographic determination of plasma free and total tazobactam and piperacillin

Gianna Di Giovamberardino, Michele Ferrannini, Gian Piero Testore, Giorgio Federici, Anna Pastore

Research output: Contribution to journalArticlepeer-review


A high-pressure liquid chromatography (HPLC) method with ultraviolet detection was developed for the measurement of plasma free and total tazobactam and piperacillin. This method is simple and fast, requiring only 11 min for the HPLC run and a sample preparation of about 11 min for total drugs and 10 min for free drugs. The procedure for the assay involves the treatment of plasma with acetonitrile for total drugs determination, and the use of a centrifugal filter device to deproteinize plasma for free drugs determination. The HPLC column, a Hypersil-ODS, was equilibrated with an eluent mixture composed of acetonitrile-potassium phosphate (pH 2.6). CVs for repeatability of tazobactam and piperacillin measurements ranged from 4.30 to 6.60; CVs for reproducibility ranged from 5.60 to 9.40. Mean analytical recoveries ranged from 100.4 to 103%. A linear relationship was obtained between peak area and drugs concentration in the range studied (0-62.5 mg/L for tazobactam and 0-500 mg/L for piperacillin). The equation for regression line were y = 19x - 1.4 for tazobactam and y = 1.7x - 0.9 for piperacillin; correlation coefficients were >0.999. The lower limit of quantitation (LLQ) for standard samples was about 0.12 mg/L for tazobactam and 0.49 mg/L for piperacillin, respectively. The lower limit of detection (LLD) was 0.06 mg/L for tazobactam and 0.24 mg/L for piperacillin. This HPLC assay for tazobactam and piperacillin is sensitive and accurate, and provides a reliable determination of both free and total tazobactam and piperacillin in human plasma, thus allowing the determination of these analytes in patients receiving tazocillin therapy.

Original languageEnglish
Pages (from-to)86-88
Number of pages3
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Issue number1-2
Publication statusPublished - Jan 1 2009


  • Assay
  • Free
  • HPLC
  • Piperacillin
  • Tazobactam
  • Total

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Clinical Biochemistry
  • Analytical Chemistry


Dive into the research topics of 'High performance liquid chromatographic determination of plasma free and total tazobactam and piperacillin'. Together they form a unique fingerprint.

Cite this