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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In: Journal of Chromatography B, Vol. 775, No. 2, 05.08.2002, p. 127-132.
Research output: Journal contributions › Journal articles › Research › peer-review
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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
JF - Journal of Chromatography B
SN - 1873-376X
IS - 2
ER -