Methodological Aspects of Measuring Human Skeletal Muscle Electrolyte Content and Ouabain Binding Capacity

The aim of the study was to evaluate the use of freeze-dried and dissected small muscle biopsy specimens (“dry”) for the determination of human muscle electrolyte content and ouabain binding capacity, compared with an easier method, without this freeze-drying step (“wet”). Freeze-drying and dissecti...

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Bibliographic Details
Published in:Analytical biochemistry 1998-07, Vol.260 (2), p.218-222
Main Authors: Djurhuus, M.Stig, Klitgaard, Niels A.H., Tveskov, Claus, Madsen, Klavs, Guldager, Bernadette, Jelnes, Rolf, Petersen, Per Hyltoft, Beck-Nielsen, Henning
Format: Article
Language:English
Subjects:
Aged
Binding Sites
Biopsy - methods
Dissection - methods
Electrolytes - analysis
Female
Freeze Drying - methods
Humans
Indicators and Reagents
Intermittent Claudication - metabolism
Intermittent Claudication - pathology
Magnesium - analysis
Male
Middle Aged
Muscle, Skeletal - chemistry
Muscle, Skeletal - metabolism
Muscle, Skeletal - pathology
Ouabain - metabolism
Potassium - analysis
Sex Characteristics
Sodium - analysis
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Summary:The aim of the study was to evaluate the use of freeze-dried and dissected small muscle biopsy specimens (“dry”) for the determination of human muscle electrolyte content and ouabain binding capacity, compared with an easier method, without this freeze-drying step (“wet”). Freeze-drying and dissection of muscle biopsy specimens reduced the variation in the determination of muscle potassium and magnesium content. The total coefficient of variation was 8.6% in the dry determination of muscle potassium content and 13.5% in the wet determination (P< 0.05). In the determination of muscle magnesium content, the total coefficient of variation was 7.4% in the dry determination and 13.7% when determined wet (P< 0.005). Muscle sodium content had a very large coefficient of variation, independent of the method used. The content of dry solids was too high in biopsies which were incubated in Tris–vanadate buffer (31.9%), compared to biopsies which were not incubated in Tris–vanadate buffer (24.9%,P< 0.001). Hereby, the measured ouabain binding capacity became too high when measured wet. In conclusion, muscle electrolyte content and ouabain binding capacity should be determined after drying and microdissection of the biopsies, because this method confers the least variation and the highest accuracy.
ISSN:0003-2697
1096-0309
DOI:10.1006/abio.1998.2625