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Glutathione disulfide variability in normal human blood
B J Mills1, J P Richie, C A Lang
1Department of Biochemistry, University of Louisville School of Medicine, Kentucky 40292.
Analytical Biochemistry
|October 1, 1994
Summary
Glutathione disulfide (GSSG) levels in human blood vary significantly due to biological differences, not just measurement errors. Optimal processing conditions reveal this true biological variation in GSSG concentrations.
Area of Science:
- Biochemistry
- Clinical Chemistry
- Human Physiology
Background:
- Prevailing opinion suggests low glutathione disulfide (GSSG) in human blood.
- Numerous studies report variable and inconsistent GSSG measurement results.
- The reasons for GSSG variability remain unclear, impacting clinical interpretation.
Purpose of the Study:
- To establish validated processing conditions for accurate GSSG measurement in human blood.
- To investigate the true biological variation of GSSG levels in normal individuals.
- To differentiate measurement artifacts from genuine physiological GSSG fluctuations.
Main Methods:
- Acid extraction of whole blood samples for GSSG analysis.
- Repetitive GSSG measurements over 6 hours to assess stability and reproducibility.
- Longitudinal GSSG measurements in normal subjects over several years.
- Comparative analysis of GSSG levels in hemolysates and ultrafiltrates.
Main Results:
- GSSG demonstrated high reproducibility and stability in acid-extracted blood over 6 hours (%CV < 10).
- Significant biological variation in GSSG levels was observed in normal subjects over time (2 to 166 nmol/10^10 RBCs; overall %CV = 46).
- Hemolysates and ultrafiltrates yielded lower GSSG values compared to optimal acid extracts.
Conclusions:
- Optimal processing conditions are crucial for accurate GSSG measurement.
- Blood GSSG concentrations exhibit substantial biological variation among individuals.
- The study highlights the importance of validated methods to reflect true physiological GSSG levels.