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Control of artifacts in plasma adenosine determinations.
1Department of Pharmacology, University of Alabama, Birmingham 35294.
Summary
Accurate measurement of plasma adenosine (ADO) is challenging due to rapid cellular uptake or production. This study developed a method to minimize these artifacts, revealing lower plasma ADO concentrations than previously reported.
Area of Science:
- Biochemistry
- Physiology
- Analytical Chemistry
Background:
- Plasma adenosine (ADO) concentration measurements vary significantly in literature.
- Discrepancies are attributed to rapid ADO uptake by red blood cells or ADO production from nucleotides during sample preparation.
Purpose of the Study:
- To establish a reliable method for determining plasma ADO concentrations in human blood.
- To identify and mitigate sources of error in plasma ADO determination.
Main Methods:
- Developed a high-performance liquid chromatography (HPLC) method for ADO determination with 3 nM sensitivity.
- Utilized a specialized stopping solution containing heparin, dilazep, EGTA, EDTA, and enzyme inhibitors.
- Collected blood in plastic tubes using plastic syringes containing the stopping solution.
Main Results:
- Plasma ADO concentrations varied widely (8.2 nM to 894 nM) depending on preparation methods.
- The optimized method yielded a mean plasma ADO concentration of 13.3 ± 1.88 nM.
- Mean ADO recovery was 78%, indicating effective inhibition of artifactual changes.
Conclusions:
- Proper blood collection and plasma preparation techniques are crucial for accurate plasma ADO measurement.
- The developed method provides a more reliable estimate of in vivo plasma ADO levels.
- Findings suggest previously reported plasma ADO concentrations may be overestimated due to methodological artifacts.