TY - JOUR

T1 - New and Rapid Fully Automated Method for Determination of Tazobactam and Piperacillin in Fatty Tissue and Serum by Column-Switching Liquid Chromatography

AU - Trittler, Rainer

AU - Ehrlich, M.

AU - Galla, T. J.

AU - Horch, R. E.

AU - Kümmerer, Klaus

PY - 2002/8/5

Y1 - 2002/8/5

N2 - 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.

AB - 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.

KW - Chemistry

KW - Piperacillin

KW - Tazobactam

UR - http://www.scopus.com/inward/record.url?scp=0037025822&partnerID=8YFLogxK

U2 - 10.1016/S1570-0232(02)00298-2

DO - 10.1016/S1570-0232(02)00298-2

M3 - Journal articles

VL - 775

SP - 127

EP - 132

JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences

SN - 1570-0232

IS - 2

ER -