New and Rapid Fully Automated Method for Determination of Tazobactam and Piperacillin in Fatty Tissue and Serum by Column-Switching Liquid Chromatography

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A sensitive and rapid HPLC assay for determining tazobactam and piperacillin in fatty tissue and serum is described. While the common methods need liquid-liquid extraction before the injection in a automated column switching HPLC, the new method works by direct injection of the filtered tissue extract or diluted serum in a automated column switching HPLC without any other pre-treatment. This was performed by the use of a NH 2-precolumn and enrichment/transfer at different pH-level. During the analyses, the NH 2-precolumn was automatically regenerated with acetonitrile-water. The chromatogram peaks for piperacillin and tazobactam were identified by the retention time and quantified by peak area. The calibration curve was linear between 1 and 16 μg/ml. The quantification limit of tazobactam was about 1 μg/ml in fatty tissue extracts and in diluted serum (calculated for pure serum 2 μg/ml), respectively. For piperacillin it was less. The described procedure allows sample clean-up and determination of the antibiotic within 35 min. The chromatograms with this easy sample treatment had the same quantity of matrix peaks and in contrast to liquid-liquid extraction no loss of piperacillin. Because of the automatically rinsing of the NH 2-precolumn during the chromatographic separation, more than 50 different biological samples could be measured with one NH 2-precolumn without loss of performance.

Original languageEnglish
JournalJournal of Chromatography B
Volume775
Issue number2
Pages (from-to)127-132
Number of pages6
DOIs
Publication statusPublished - 05.08.2002
Externally publishedYes

    Research areas

  • Chemistry - Piperacillin, Tazobactam

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