Comparison of three different protocols for obtaining hemolysis

Nora Nikolac Gabaj; Marijana Miler; Alen Vrtaric; Ivana Celap; Marina Bocan; Petra Filipi; Vanja Radisic Biljak; Ana-Maria Simundic; Vesna Supak Smolcic; Marija Kocijancic

doi:10.1515/cclm-2021-1227

Article

Comparison of three different protocols for obtaining hemolysis

Nora Nikolac Gabaj , Marijana Miler , Alen Vrtaric , Ivana Celap , Marina Bocan , Petra Filipi , Vanja Radisic Biljak , Ana-Maria Simundic , Vesna Supak Smolcic and Marija Kocijancic

Published/Copyright: February 23, 2022

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From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 60 Issue 5

Abstract

Objectives

Hemolysis is associated with erroneous or delayed results. Objectives of the study were to compare four different methods for obtaining hemolysis in vitro on three different analyzers.

Methods

Hemolysis was prepared with addition of pure hemoglobin into serum pool, osmotic shock, aspiration through blood collection needle, freezing/thawing of whole blood. Biochemistry parameters were measured in duplicate at Architect c8000 (Abbott, Abbott Park, USA), Beckman Coulter AU680 (Beckman Coulter, Brea, USA) and Cobas 6000 c501 (Roche, Mannheim, Germany), according to manufacturers’ declarations. Cut-off value was defined as the highest value of H index with corresponding bias lower than acceptance criteria.

Results

We were not able to obtain results with freezing protocol. On all three platforms, lowest number of analytes were sensitive to hemolysis at H=0.5 using method of adding free hemoglobin. When osmotic shock was used, cut-off values for the most analytes were generally met at lower values. Hemolysis significantly interfered with measurement of potassium and lactate dehydrogenase (LD) at H=0.5 on all platforms. The most of the tested analytes had the lowest acceptable H index when aspiration method was used. At the low level of hemolysis (H=0.8) glucose, sodium, potassium, chloride, phosphate, and LD were affected on all analyzers, with some additional analytes depending on the manufacturer.

Conclusions

Hemolysis interference differs on different analyzers and according to protocol for obtaining hemolysis. Aspiration method was generally the most sensitive to hemolysis interference, while addition of free Hb was the most resistant.

Keywords: hemolysis; interference; osmotic shock; preanalytical errors

Corresponding author: Marijana Miler, Working Group for Preanalytical Phase of the Croatian Society of Medical Biochemistry and Laboratory Medicine, Zagreb, Croatia; and Department of Clinical Chemistry, Sestre milosrdnice University Hospital Center, Zagreb, Croatia, E-mail: marijana.miler@gmail.com

Research funding: None declared.
Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.
Competing interests: Authors state no conflict of interest.
Informed consent: Not applicable.
Ethical approval: The study was approved by the Ethical Committee of the Sestre milosrdnice University Hospital Center. For the study, samples were prepared from the remaining portions of serum and whole blood pools after routine laboratory testing was completed. Demographic, anamnestic or clinical data on patients were not collected.

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Received: 2021-11-23

Accepted: 2022-02-11

Published Online: 2022-02-23

Published in Print: 2022-04-26

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Articles in the same Issue

https://doi.org/10.1515/cclm-2021-1227

Keywords for this article

hemolysis; interference; osmotic shock; preanalytical errors