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Development of an automatic solid phase extraction and liquid chromatography mass spectrometry method by using a monolithic column for the analysis of Cyclosporin A in human plasma
Authors:Covadonga Alvarez
Institution:a Department of Pharmaceutical Technology, Faculty of Pharmacy, Complutense University of Madrid, 28040 Madrid, Spain
b Laboratory of Bioanalytical Chemistry, Gerontology Research Center, National Institutes of Health, 5600 Nathan Shock Drive, Baltimore, MD 21224-6825, USA
Abstract:A sensitive and specific and automated liquid chromatography-electrospray mass spectrometric (LC-ESI-MS) assay for the quantification of Cyclosporin A in human plasma was developed. Following a simple protein precipitation step, the supernatant was extracted on-line and directly injected into the system LC-ESI-MS. A relatively new type of monolithic column consisting of a silica rod with bimodal pore structure was used to achieve a retention time of 2.4 min with a very low backpressure at a flow rate of 1 ml/min. The assay was linear from 0.050 to 1.000 μg/ml. The mean recovery was 91%. The mean inter-day and intra-day precisions were 1.85% and 2.83%, respectively. The combination of the automated solid phase extraction and the low retention time achieved with this columns increase the throughput and decrease the time of analysis of each sample. This technology is useful in order to improve the efficiency of the bioanalytical studies.
Keywords:Cyclosporin A  Monolithic columns  LC-ESI-MS
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