Reproducibly Simulating Hemolysis, for Evaluating Its Interference with Chemical Methods

Meites, Samuel

doi:10.1093/clinchem/19.11.1319a

Cited by 20 publications

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“…5,9,10 Paired interference testing using the osmotic shock haemolysate protocol, adapted by the CLSI, 6 was developed to study analytical interference from 'dissolved haemoglobin'. 8 In the protocol, therefore, dilution with DIW while preparing haemolysate was not considered of importance. 8 For parameters like sodium and chloride, however, which are under very tight physiological control, 18 even small differences in dilution could be of significance.…”

Section: Discussionmentioning

confidence: 99%

“…8 In the protocol, therefore, dilution with DIW while preparing haemolysate was not considered of importance. 8 For parameters like sodium and chloride, however, which are under very tight physiological control, 18 even small differences in dilution could be of significance.…”

Section: Discussionmentioning

confidence: 99%

“…There are several well-described methods to study in vitro haemolysis and most utilise osmotic shock, freezethaw, shear stress or a combination of these to lyse cells. 3 Most widely used has been the osmotic shock protocol described by Meites 8 which has been adapted by CLSI and included in the CLSI guidance document for interference testing in Clinical Chemistry. 6 Data on the effect of haemolysis on plasma sodium and chloride concentrations are conflicting.…”

Section: Introductionmentioning

confidence: 99%

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Conflicting effects of haemolysis on plasma sodium and chloride are due to different haemolysis study protocols: A case for standardisation

et al. 2021

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Background Haemolysis has been reported as having a positive, negative or no effect on plasma sodium (PNa) and chloride (PCl). We investigated the haemoltytic effect of different haemolysis protocols on PNa and PCl using modelling and laboratory experiments. Methods In a modelling experiment, percentage change and recovery due to dilution in routinely ( in vitro) haemolysed samples were compared against shear stress haemolysis and samples spiked with haemolysate from whole blood freeze–thaw, packed cells freeze–thaw and osmotic shock protocols. The results were compared against a control base pool. Additionally, for the osmotic shock method, results were compared against saline- and deionised water (DIW)-spiked controls. In a laboratory experiment, percentage change and recovery were similarly compared using haemolysate from whole blood freeze–thaw and osmotic shock protocols. PNa, PCl and H-index were measured on the Abbott Architect and haemoglobin on the Sysmex XN-9000. Results In the modelling experiment, the percentage decrease in PNa and PCl was similar in in vitro haemolysis, shear stress haemolysis, whole blood freeze–thaw haemolysis and packed cells freeze–thaw haemolysis and this was lower compared to the osmotic shock method. In the laboratory experiment, the change in PNa compared to the base pool was less ( p < 0.001) per unit increase in H-index in the freeze–thaw method (−0.33 mmol, 95% CI −0.35 to −0.31) compared to the osmotic shock method (−0.65 mmol, 95% CI −0.66 to −0.64). PCl did not change with haemolysis in the freeze–thaw method and changed by −0.21 ± 0.01 mmol per unit increase in the H-index in the osmotic shock method. Recovery of PNa and PCl increased with increasing H-index in both methods. Conclusion The osmotic shock protocol is inappropriate for haemolysis studies because of dilution with DIW used for cell lysis. Recovery calculations may incorrectly compensate for genuine dilution caused by haemolysis.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Conflicting effects of haemolysis on plasma sodium and chloride are due to different haemolysis study protocols: A case for standardisation

et al. 2021

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“…Also, we used the freeze-thaw method to achieve full lysis of the isolated cells since the main objective was to evaluate the relationship between the concentration of the lysed cells and the analyte. Supported by Lippi’s article and evaluating that Meites has stressed that the serum dilution ratio should be kept at minimum (not more than 5%) in interference experiments, the sample matrix of this study was preserved by using the ratio of 1/20 for the hemolysate (28, 29).…”

Section: Discussionmentioning

confidence: 99%

Effect of Blood Cell Subtypes Lysis on Routine Biochemical Tests

Ünlü¹,

Küme²,

Emek³

et al. 2018

Journal of Medical Biochemistry

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SummaryBackground: The aim of this study is to establish the contribution of blood cells subtypes on hemolysis. Methods: Separated blood cell subtype suspensions prepared with blood from 10 volunteers were serially diluted to obtain different concentrations of cell suspensions. The cells were fully lysed and cell hemolysates were added (1:20) to aliquots of serum pool. Thus, seven serum pools with different concentrations of interferent were obtained for each blood cell subtype. Biochemical parameters and serum indices were measured by an autoanalyzer. As cell lysis markers, free hemoglobin was measured by spectrophotometry while myeloperoxidase and b-thromboglobulin were measured by enzyme immunoassay. The percent changes in analyte levels of the serum pools were evaulated by Wilcoxon Signed Rank Test and compared with clinical thresholds defined for each test. Results: The clinical thresholds were exceeded in lactate dehydrogenase, potassium, aspartate aminotransferase, creatine kinase, magnesium, total protein, total cholesterol, inorganic phosphate, glucose for red blood cells (RBC); lactate dehydrogenase, aspartate aminotransferase, total protein, inorganic phosphate and glucose for platelets (PLT). Free hemoglobin was significantly correlated with RBC (r=0.999; p=0.001), while myeloperoxidase and b thromboglobulin showed no significant correlation to white blood cells (WBC) and PLT, respectively. Conclusions: The effect of RBC hemolysis in serum on the routine biochemical tests are clearly established, yet, additional studies are required in order to verify this kind of effects of PLT and WBC hemolysis.

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“…Petricoin and Liotta omitted the extensive historic role of MS in clinical chemistry, including the numerous contributions and advancements in the previous two decades. Bruns et al accurately describe the recent and important contributions of mass spectrometry and included timely manuscripts in the text, Molecular Testing in Laboratory Medicine (2 ). Dozens of other articles have appeared in Clinical Chemistry and other journals regarding MS applications in clinical chemistry, as evidenced in surveys of the literature that I and others have written recently (3,4 ).…”

Section: Andries J Bakkermentioning

confidence: 99%

Hemolysis Interference in the Ortho-Clinical Diagnostics Vitros ECi cTnI Assay

Hawkins

2003

Clinical Chemistry

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Assay specificity is an important requirement for cardiac troponin measurement (1). This study examines the effect of hemolysis on two troponin I (cTnI) assays: the Ortho-Clinical Diagnostics Vitros ® ECi and the Beckman-Coulter Access 2 (Ac-cuTnI). Erythrocyte hemolysates were prepared according to the procedure of Meites (2). Aliquots were added to unhemolyzed, heparinized plasma pools with three different cTnI concentrations (for the ECi, 0.05, 0.1, and 0.3 g/L; for the AccuTnI, 0.1, 0.2, and 0.5 g/L) to produce severely hemolyzed specimens. Each of these specimens was diluted in the respective unhemolyzed plasma pool to form nine samples. The hemolysis index (HI; Roche 917 analyzer) and cTnI (ECi and Access) were measured in all samples. The magnitude of hemolysis interference varied with the cTnI concentration. Multiple linear regression using the HI and measured cTnI in the hemolyzed sample as variables to predict true cTnI concentration (the original pool cTnI concentration) was performed with Sigmastat 2.0 (Jandel Corporation). A single equation (r ϭ 0.91) was sufficient for the Access, whereas two equations (r ϭ 0.90 and 0.92) covering different HI ranges were needed for the ECi.

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Reproducibly Simulating Hemolysis, for Evaluating Its Interference with Chemical Methods

Cited by 20 publications

References 0 publications

Conflicting effects of haemolysis on plasma sodium and chloride are due to different haemolysis study protocols: A case for standardisation

Conflicting effects of haemolysis on plasma sodium and chloride are due to different haemolysis study protocols: A case for standardisation

Effect of Blood Cell Subtypes Lysis on Routine Biochemical Tests

Hemolysis Interference in the Ortho-Clinical Diagnostics Vitros ECi cTnI Assay

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