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Analysis of microsomal metabolic stability using high-flow-rate extraction coupled to capillary liquid chromatography-mass spectrometry
Uppsala University, Teknisk-naturvetenskapliga vetenskapsområdet, Chemistry, Department of Chemistry. Analytisk kemi.
Uppsala University, Teknisk-naturvetenskapliga vetenskapsområdet, Chemistry, Department of Chemistry. Analytisk kemi.
2004 (English)In: Journal of Chromatography B, no 806, 119-126 p.Article in journal (Refereed) Published
Abstract [en]

A method described for on-line high-speed extraction of microsomal samples and analysis by capillary liquid chromatography - mass spectrometry (LC-MS) for the determination of metabolic stability in connection with the development of positron emission tomography (PET) tracers. the method allowed direct injections of large sample volumes at a fast extraction rate, providing a gain in both sensitivity and sample preparation time. The calibration curve of the test compound flumazenil (Ro 15-1788) was linear in the concentration range of 1-150 nM, with a correlation coefficient exceeding 0.999. The accuracy of the method ranged from 98 to 101%. A high precision was obtained, with mean intra-assay and inter-assay relative standard deviations of at most 1.4 and 1.5%, respectively, for quality control (QC) samples. The extraction efficiency was determined to be 99.4%, the total recovery 96% and the carryover to <0.23%. Extractions were performed in a concentration interval of 30-300 nM without any sign of column overload. The method was successfully used for determining the microsomal metabolic stability of flumazenil. As a result, the described analysis system is currently used for metabolic screening of PET tracer candidates in our laboratory.

Place, publisher, year, edition, pages
2004. no 806, 119-126 p.
Keyword [en]
High-flow-rate extraction, Metabolic stability, Positron emission tomography
Identifiers
URN: urn:nbn:se:uu:diva-70671OAI: oai:DiVA.org:uu-70671DiVA: diva2:98582
Available from: 2005-04-29 Created: 2005-04-29 Last updated: 2011-01-12

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