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The effect of centrifugation speed and time on pre-analytical platelet activation

  • Anna C. Söderström EMAIL logo , Mads Nybo , Christian Nielsen and Pernille J. Vinholt
Published/Copyright: May 26, 2016
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 54 Issue 12

Abstract

Background:

The results of laboratory analyses are affected by pre-analytical variables, and in particular can platelets be activated by shear handling stress and secrete granular substances. We therefore evaluated the effect of centrifugation speed and time on pre-analytical platelet activation.

Methods:

Citrate- and EDTA-anticoagulated blood from healthy volunteers were centrifuged at 80–10,000 g for 5–15 min to prepare plasma and platelet-rich plasma. Pre-analytical platelet activation was assessed by flow cytometric measurement of platelet P-selectin (CD62p) expression. Blood cell counts, mean platelet volume (MPV), immature platelet fraction (IPF), and platelet distribution width (PDW) were measured. Platelet aggregation in platelet-rich plasma induced by arachidonic acid (AA), ADP or thrombin receptor activator peptide-6 (TRAP) was tested by 96-well aggregometry.

Results:

The median percentage of platelets expressing P-selectin in citrate- and EDTA-plasma centrifuged at 2000 g for 10 min were 43% [interquartile range (IQR), 38%–53%] and 56% (IQR, 31%–78%), respectively (p=0.82). Platelet-rich plasma prepared at 100–250 g for 10 min had significantly lower platelet P-selectin expression (11%–15%), p<0.001. Platelet count in plasma samples decreased with increasing speed but platelets were only completely removed if plasma was re-centrifuged. In platelet-rich plasma, increasing centrifugation speed significantly increased platelet yield but decreased contamination from other blood cells, platelet composition was altered as platelet parameters (MPV, IPF, and PDW) was lowered. Platelet aggregation was not affected by the centrifugation speed platelet-rich plasma was prepared.

Conclusions:

Proportional to centrifugation speed, platelets in plasma and platelet-rich plasma were activated with centrifugation speed, cell content and composition changed while platelet aggregation was unaltered.

Keywords: centrifugation; plasma; platelet aggregation; platelets; P-selectin

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Received: 2016-1-28
Accepted: 2016-4-25
Published Online: 2016-5-26
Published in Print: 2016-12-1

©2016 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2016-0079
Keywords for this article
centrifugation; plasma; platelet aggregation; platelets; P-selectin

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Biomarkers, inflammation and cancer: where to go?
  4. Review
  5. Analysis, detection and quantitation of mixed cryoglobulins in HCV infection: brief review and case examples
  6. Mini Reviews
  7. Calcitonin measurement and immunoassay interference: a case report and literature review
  8. Exosomal non-coding RNAs: a promising cancer biomarker
  9. Opinion Paper
  10. Towards a new paradigm in laboratory medicine: the five rights
  11. EFLM Recommendation
  12. Recommendation for the review of biological reference intervals in medical laboratories
  13. IFCC Position Paper
  14. Assuring the quality of interpretative comments in clinical chemistry
  15. General Clinical Chemistry and Laboratory Medicine
  16. The effect of centrifugation speed and time on pre-analytical platelet activation
  17. An assessment of clinical laboratory performance for the determination of manganese in blood and urine
  18. Evaluation of antiphospholipid antibody assays using latent class analysis to address the lack of a reference standard
  19. Evaluation of the hypochromic erythrocyte and reticulocyte hemoglobin content provided by the Sysmex XE-5000 analyzer in diagnosis of iron deficiency erythropoiesis
  20. Cancer Diagnosis
  21. Identifying risk in the use of tumor markers to improve patient safety
  22. Association between Echinococcus granulosus infection and cancer risk – a pilot study in Cyprus
  23. Dynamic change of the systemic immune inflammation index predicts the prognosis of patients with hepatocellular carcinoma after curative resection
  24. Aberrant methylation of tumour suppressor genes WT1, GATA5 and PAX5 in hepatocellular carcinoma
  25. Cardiovascular Diseases
  26. High homocysteine and low folate plasma concentrations are associated with cardiovascular events but not bleeding during warfarin treatment
  27. Hemolysis and Coagulation
  28. Hemolysis rates in blood samples: differences between blood collected by clinicians and nurses and the effect of phlebotomy training
  29. Letters to the Editor
  30. Harmonisation of the laboratory testing process: need for a coordinated approach
  31. Pseudohyperkalemia due to severe leukocytosis: case presentation
  32. When obtaining a blood sample from the right arm was not the right thing to do: a case of elevated parathyroid hormone levels 27 years after thyroidectomy
  33. Raising awareness of assay compatibility with heparinized plasma
  34. Improved protocol for extraction of genomic DNA from formalin-fixed paraffin-embedded tissue samples without the use of xylene
  35. Effect of refrigeration, centrifugation, acidification, heat treatment and storage on urine calcium, magnesium and phosphate
  36. Letter in response to: Identifying risk in the use of tumor markers to improve patient safety
  37. Letter to the Editor in reply to Dayyani and Morgenstern’s comment on the article “Identifying risk in the use of tumor markers to improve patient safety”
  38. Level of red cell distribution width is affected by various factors
  39. Red blood cell distribution: an index without additional cost in estimating the prognosis of acute pancreatitis
  40. Testing of total 25(OH)vitamin D: agreement and discrepant cases between Cobas® 8000 and Liaison® XL methods
  41. Expression profiling and ontology analysis of circulating long non-coding RNAs in septic acute kidney injury patients
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