College of Veterinary Medicine

Hemolysis in Blood Specimens

Hemolysis is the breaking down of red blood cells due to the mishandling of blood samples during routine blood collection and transport.

Serum from heavily hemolyzed blood specimens can interfere with the antigen-antibody reaction and color measurement in immunoassays such as ELISAs. Trace Mineral, Serum testing is also affected by hemolysis resulting in increased iron, magnesium, and potassium concentrations, while possibly decreasing those of sodium.

The accuracy of the test results is dependent on the quality of the specimens. A quality serum sample for diagnostic testing can be obtained by following best practices during collection, handling, and transporting the samples to the diagnostic laboratory.

Best Practices to Prevent Hemolysis

Collection and Handling

  1. Choose the right gauge needle.
  2. Allow the alcohol used for cleansing the venipuncture site to dry completely before drawing the blood.
  3. After drawing the blood, remove the needle from the syringe and transfer the blood gently down the side of the collection tube (serum separator tube (Tiger top tubes) or red top tubes without anticoagulants).
  4. Invert the tube gently as recommended by the tube manufacturer.
  5. Next, place the tube upright for 15-30 minutes at room temperature until clot formation is complete.
  6. Centrifuge the sample and pour the serum into a sterile tube (ex: red top tube). Store at ~4°C (refrigeration temperature) until ready to ship to the lab.
  • DO NOT freeze the clotted sample.
  • If you do not have access to a centrifuge, a clotted sample can be submitted.

Packaging for the Shipment

  1. Use proper packaging using sufficient foam or bubble wrap to prevent specimens from shifting during shipment.
  2. Ship the samples in an insulated container with a cold pack for overnight delivery.

For more information on the shipment, please refer to:

Hemolysis Reference Palette Guide

Hemolysis_Palette_Bookmark-P (PDF)

References

  1. Effects of blood sample mishandling on ELISA results for infectious bronchitis virus, avian encephalomyelitis virus and chicken anaemia virus
  2. Interferences in Immunoassay
  3. Interferences in Immunoassay journals.sagepub.com
  4. Jugular Venepuncture – Get the best sample with these tips for VNs
  5. A Quick-Reference Tool for Hemolysis Status
  6. Transmission of Blood and Serum Samples
  7. Plasma and Serum Preparation Protocols - Thermofisher
  8. Influence of Hemolysis on Routine Clinical Chemistry Testing