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Related Experiment Videos

Homocysteine export from erythrocytes and its implication for plasma sampling.

A Andersson1, A Isaksson, B Hultberg

  • 1Department of Clinical Chemistry, University Hospital, Lund, Sweden.

Clinical Chemistry
|July 1, 1992
PubMed
Summary

Storing blood samples uncentrifuged causes plasma homocysteine to increase, especially at higher temperatures. Erythrocytes are the primary source, highlighting the need for strict blood sampling protocols for accurate homocysteine assays.

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Area of Science:

  • Biochemistry
  • Clinical Chemistry
  • Hematology

Background:

  • Plasma homocysteine levels are critical biomarkers in various health conditions.
  • Accurate measurement of homocysteine requires stable blood sample integrity post-collection.
  • Pre-analytical variables, such as sample handling, can significantly impact assay results.

Purpose of the Study:

  • To investigate the impact of uncentrifuged blood storage on plasma homocysteine concentrations.
  • To determine the temperature dependence of homocysteine increase during blood storage.
  • To identify the cellular components responsible for homocysteine generation in stored blood.

Main Methods:

  • Blood samples were stored uncentrifuged at different temperatures (e.g., 4°C and 37°C).

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  • Concentrations of free and total homocysteine, glutathione, cysteinylglycine, gamma-glutamylcysteine, and cysteine were measured over time.
  • Erythrocytes were investigated as a potential source of homocysteine increase.

    • Main Results:

    • Plasma homocysteine (free and total) significantly increased in uncentrifuged blood over time.
    • The rate of increase was temperature-dependent, with a maximal increase of 3.0 µmol/L/h for total homocysteine at 37°C.
    • Erythrocytes were identified as the main contributors to the observed increase in plasma homocysteine.
    • Plasma glutathione, cysteinylglycine, and gamma-glutamylcysteine increased, while cysteine decreased during storage.

    Conclusions:

    • Uncentrifuged blood storage leads to artifactual increases in plasma homocysteine, particularly the free fraction.
    • Erythrocytes appear to be the source of increased homocysteine, likely through adenosylmethionine-dependent protein carboxymethylations.
    • Strict adherence to pre-analytical sampling conditions is essential for reliable plasma homocysteine and free homocysteine measurements.