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Estimation of plasma serotonin using isopycnic centrifugation
Harm J. Gijsman1, Robbert-Jan Verkes, Jolanda C. M. Schouten-Verhagen
1Centre for Human Drug Research, Leiden, The Netherlands.
Human Psychopharmacology
|October 31, 2002
Summary
Accurate serotonin (5-HT) measurement in plasma is crucial. A new isopycnic method using Percoll density gradients significantly reduces platelet contamination, yielding more reliable 5-HT levels for platelet research.
Area of Science:
- Biochemistry
- Neuroscience
- Cell Biology
Background:
- Accurate measurement of plasma serotonin (5-HT) is essential for using platelets as a model for serotonergic neurons.
- Assay validity is compromised by 5-HT release from residual platelets during sample preparation.
Purpose of the Study:
- To evaluate a novel isopycnic method for separating cells from plasma.
- To compare the isopycnic method with traditional differential centrifugation for plasma 5-HT analysis.
Main Methods:
- Developed an isopycnic method involving Percoll density gradients and centrifugation.
- Added whole blood to a diluted Percoll gradient for gentle and complete cell separation.
- Compared results with the standard differential centrifugation technique.
Main Results:
- The isopycnic method yielded plasma 5-HT levels nine times lower on average compared to differential centrifugation.
- The reduction in 5-HT levels was directly correlated with the number of residual platelets.
- Intra-individual variation in 5-HT measurements decreased threefold with the isopycnic method.
Conclusions:
- The isopycnic method provides a more precise and accurate estimation of plasma 5-HT levels.
- Utilizing Percoll density gradients effectively minimizes platelet interference in plasma serotonin assays.
- This improved method enhances the reliability of platelet-based models for serotonergic neuron studies.
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