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 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.
Djurhuus
M S
MS
Department of Clinical Biochemistry and Genetics, Odense University Hospital, Odense C, 5000, Denmark.
Klitgaard
N A
NA
Tveskov
C
C
Madsen
K
K
Guldager
B
B
Jelnes
R
R
Petersen
P H
PH
Beck-Nielsen
H
H
D000368:Aged; D001665:Binding Sites; D001706:Biopsy / Q000379:methods; D004210:Dissection / Q000379:methods; D004573:Electrolytes / Q000032:analysis*; D005260:Female; D005612:Freeze Drying / Q000379:methods; D006801:Humans; D007202:Indicators and Reagents; D007383:Intermittent Claudication / Q000378:metabolism* / Q000473:pathology; D008274:Magnesium / Q000032:analysis; D008297:Male; D008875:Middle Aged; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000378:metabolism* / Q000473:pathology; D010042:Ouabain / Q000378:metabolism*; D011188:Potassium / Q000032:analysis; D012727:Sex Characteristics; D012964:Sodium / Q000032:analysis
D004573:Electrolytes; D007202:Indicators and Reagents; D010042:Ouabain; D012964:Sodium; D008274:Magnesium; D011188:Potassium