Method for measuring high energy phosphates in fragments of human muscle fibres

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This research publication describes a method developed in the Amsterdam Department of Anthony Sargeant. The data was collected by an outstanding Greek PhD student, Christina Karatzaferi, directed by Tony and his colleague Arnold de Haan.
Journal of Chromotography
J Chromatogr B Biomed Sci Appl. 1999 Jul 9;730(2):183-91

Abstract A sensitive and reproducible method for the determination of adenine nucleotides (ATP, IMP) and creatine compounds [creatine (Cr), phosphocreatine (PCr)] in freeze-dried single human muscle fibre fragments is presented. The method uses isocratic reversed-phase high-performance liquid chromatography of methanol extracts. Average retention times (min) of ATP, IMP and PCr, Cr from standard solutions were 10.6+/-0.42, 2.11+/-0.06 (n=6) and 10.5+/-0.31 and 1.19+/-0.02 (n=9), respectively. Detection limits in extracts from muscle fibre fragments were 2.0, 1.0, 3.0 and 2.0 mmol/kg dm, respectively. The assay was found successful for analysis of fibre-fragments weighing > or = 1 microg.

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