New sensitive assay of vancomycin in human plasma using high-performance liquid chromatography and electrochemical detection |
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Authors: | P. Favetta J. Guitton N. Bleyzac C. Dufresne J. Bureau |
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Affiliation: | a Service de Pharmacie, Hôpital Lyon-Sud – Secteur Ste Eugénie, 165 Chemin du Grand Revoyet, 69310 Pierre-Bénite, France;b Fédération de Biochimie, Hôpital Edouard Herriot, Place d’Arsonval, 69437 Lyon Cedex 03, France;c Laboratoire de Physiologie Rénale et Métabolique, INSERM U499, Faculté de Médecine Laennec, 8 Rue G. Paradin, 69008 Lyon, France;d Service de Pharmacie, Hôpital Debrousse, 29 Rue Sœur Bouvier, 69322 Lyon Cedex 05, France;e Département de Pharmacie Clinique, ISPBL, 8 Avenue Rockefeller, 69373 Lyon Cedex 08, France |
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Abstract: | A method using reversed-phase high-performance liquid chromatography with electrochemical detection for the analysis of vancomycin in human plasma was developed. Chromatographic conditions included an octadecyl column, a mobile phase of acetonitrile–sodium phosphate buffer (pH 7) (12:88), a total run time of 12 min, and coulometric electrochemical detection at +700 mV. Linear detector response was found in the range 5–100 μg ml−1 after a 1:80 dilution or from 0.5 to 50 μg ml−1 after a 1:20 dilution of the samples. In both cases the correlation coefficient (r) of the calibration curve standard was better than 0.995. Vancomycin determination was based on a denaturation of plasma proteins with methanol, then a dilution with mobile phase was performed. Recovery of vancomycin from plasma was 103.1±3.9%, and no interference from commonly used drugs or endogenous compounds was observed. A significant correlation was shown with the EMIT assay (r=0.92, P<0.001) using clinical samples from children. This HPLC technique is simple, sensitive, rapid, precise, selective and requires only 100 μl of plasma for completion. |
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Keywords: | Vancomycin |
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