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Updated: Jul 12, 2026

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
Basic extraction studies of benzylpenicillin and its determination by liquid chromatography with pre-column
1Swedish University of Agricultural Sciences, Faculty of Veterinary Medicine, Department of Pharmacology and Toxicology, Uppsala Biomedical Centre.
Abstract:
In order to develop a sensitive and precise chemical bioanalysis of benzylpenicillin in biological fluids, basic studies on benzylpenicillin are presented. These studies include pH dependent stability, extraction in aqueous ethylacetate systems in different pH and buffer compositions. A pre-column derivatisation LC method for the detection of benzylpenicillin also is described. The derivatisation of the sample is performed by the beta-lactam ring specific formation of the mercuric(II)mercaptide of the penicillenic acid in the presence of imidazole. The UV spectral properties of the derivative are utilised for detection. The chromatographic conditions are optimised with reference to the pH, the methanol, the imidazole and the mercury(II)chloride content of the mobile phase as well as the column temperature.
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