Analytical and Clinical Interference of Sample Hemolysis in Evaluating Blood Biochemical and Endocrine Parameters in Cows

Author:

Kovačević Dražen1,Cincović Marko1,Majkić Mira1,Spasojević Jovan1ORCID,Djoković Radojica2,Nikolić Sandra1,Došenović Marinković Maja3,Delić Vujanović Biljana3,Obradović Nemanja4,Anđušić Ljiljana5,Čukić Aleksandar5ORCID,Petrović Miloš2ORCID,Starič Jože6ORCID,Ježek Jožica6

Affiliation:

1. Faculty of Agriculture, University of Novi Sad, Square Dositeja Obradovića 7, 21000 Novi Sad, Serbia

2. Faculty of Agronomy, University of Kragujevac, Cara Dušana 34, 32000 Čačak, Serbia

3. Academy of Applied Studies Šabac, Dobropoljska 5, 15000 Šabac, Serbia

4. Pasteur Institute Novi Sad, Hajduk Veljkova 1, 21000 Novi Sad, Serbia

5. Faculty of Agriculture, University of Priština, Kopaonička bb, 38219 Lešak, Serbia

6. Veterinary Faculty, University of Ljubljana, Gerbičeva 60, 1000 Ljubljana, Slovenia

Abstract

Hemolysis is a common cause of errors in laboratory tests as it affects blood parameters and leads to a positive or negative bias. This study aims to examine the relationship between the level of hemolysis (expressed as cell-free hemoglobin concentration, g/L) and the variability of metabolic and endocrine parameters and to determine the threshold level of hemolysis that causes an analytically and clinically significant bias for the twenty most frequently examined blood parameters in cows. Paired blood samples of 10 mL each were obtained from 30 cows. One was subjected to mechanical trauma and plasma was extracted directly from the other. Hemolyzed and non-hemolyzed samples from the same animal were mixed to obtain final samples with cell-free hemoglobin concentrations of 0, 1, 2, 4, 6, 8, and 10 g/L. Metabolic and endocrine parameters were measured in the samples and their deviation and the linear equation between the level of hemolysis and the deviation were determined. The following threshold values of hemolysis were determined, which correspond to the acceptable analytical (lower value) and clinical (upper value) levels of parameter variability: BHB 0.96 and 4.81; NEFA 0.39 and 3.31; GLU 0.38 and 3.90; ALB 1.12 and 6.11; TPROT 1.40 and 6.80; UREA 6.62 and 20.1; TBIL 0.75 and 5.65; AST 0.11 and 2.18; GGT 1.71 and 8.90, LDH 0.01 and 0.11, ALP 0.97 and 2.95; TGC 1.56 and 15.5; CHOL 1.29 and 8.56; Ca 5.68 and 25.7; P 0.57 and 8.43; Mg 1.10 and 8.47; INS 1.15 and 3.89; T3 8.19 and 15.6; T4 8.97 and 18.5; and CORT 2.78 and 11.22 g/L cell-free hemoglobin. Three decision levels are available for each metabolic and endocrine parameter: if hemolysis is below the lower (analytical) threshold value, results can be reported without restriction; if hemolysis is between the lower and upper thresholds, the results can be issued with guidance in the form of corrective linear equations; and if hemolysis is above the upper (clinical) threshold, the results and sample must be discarded. This method contributes to an optimal approach to hemolysis interference with metabolic profile parameters in blood samples from cows.

Funder

Provincial Secretariat for Higher Education and Scientific Research of Vojvodina

Slovenian Research and Innovation Agency

Publisher

MDPI AG

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