Effects of hemolysis interference on routine biochemistry parameters
ABSTRACT Hemolysis is still the most common reason for rejecting samples, while reobtaining a new sample is an important problem. The aim of this study was to investigate the effects of hemolysis in different hemolysis levels for mostly used biochemical parameters to prevent unnecessary rejections.
Sixteen healthy volunteers were enrolled in the study. Four hemolysis levels were constituted according to hemoglobin concentrations and they were divided into five groups: Group I: 0-0.10 g/L, Group II:0.10-0.50 g/L, Group III: 0.51-1.00 g/L, Group IV: 1.01-2.50 g/L, Group V: 2.51-4.50 g/L. Lysis was achieved by mechanical trauma.
Hemolysis interference affected lactate dehydrogenase (LD) and aspartate aminotransferase (AST) almost at undetectable hemolysis by visual inspection (plasma hemoglobin < 0.5 g/L). Clinically meaningful variations of potassium and total bilirubin were observed in moderately hemolyzed samples (hemoglobin > 1 g/L). Alanine aminotransferase (ALT), cholesterol, gamma glutamyltransferase (GGT), and inorganic phosphate (P) concentrations were not interfered up to severely hemolyzed levels (hemoglobin: 2.5-4.5 g/L). Albumin, alkaline phosphatase (ALP), amylase, chloride, HDL-cholesterol, creatine kinase (CK), glucose, magnesium, total protein, triglycerides, unsaturated iron binding capacity (UIBC) and uric acid differences were statistically significant, but remained within the CLIA limits.
To avoid preanalytical visual inspection for hemolysis detection, improper sample rejection, and/or rerun because of hemolysis, it is recommended in this study that, routine determination of plasma or serum free hemoglobin concentrations is important. For the analytes interfered with hemolysis, new samples have to be requested.
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ABSTRACT: Researchers analyse hormones to draw conclusions from changes in hormone concentrations observed under specific physiological conditions and to elucidate mechanisms underlying their biological variability. It is, however, frequently overlooked that also circumstances occurring after collection of biological samples can significantly affect the hormone concentrations measured, owing to analytical and pre-analytical variability. Whereas the awareness for such potential confounders is increasing in human laboratory medicine, there is sometimes limited consensus about the control of these factors in rodent studies. In this guide, we demonstrate how such factors can affect reliability and consequent interpretation of the data from immunoassay measurements of circulating metabolic hormones in rodent studies. We also compare the knowledge about such factors in rodent studies to recent recommendations established for biomarker studies in humans and give specific practical recommendations for the control of pre-analytical conditions in metabolic studies in rodents.12/2012; 1(1-2):47-60. DOI:10.1016/j.molmet.2012.07.004
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ABSTRACT: Corresponding author. e-mail: email@example.com. Hemolysis is still the most common reason for rejecting samples, while reobtaining a new sample is an important problem. The aim of this study was to explain the decrease in the activity of Alkaline phosphatase (ALP) enzyme after hemolysis of blood samples and whether conversion of zinc and magnesium ions to inhibit alkaline phosphatase activity after they released from red blood cells. Twenty healthy volunteers were enrolled in the study. Four hemolysis levels were constituted according to hemoglobin concentrations (0.02, 0.27 ,0.75 and 3.34 g/L). Non–hemolysed samples was obtained from each volunteer and considered as control. Hemolysis was achieved by mechanical trauma. Alkaline phosphatase activity and the concentrations of zinc and magnesium ions were measured in the hemolysed and non-hemolysed samples. Ten non-hemolysed serum samples (Hb concentration was < 0.02 g/L) were divided into two groups samples A and B. ALP activity was measured in these samples. In vitro study was carried out including the addition 68.3 mg/dL of magnesium chloride to group A and 5.1 µg/dL of zinc chloride to group B. ALP activity was measured in the sera of the two groups. The significant decrease (p<0.001) in ALP activity was observed at moderate (13.2 ± 7.2IU/L), and severe hemolysis (5.5± 2.3IU/L) compared with that in non hemolysed samples. In these levels of hemolysis the concentrations of Zn +2 ions (5.1 ±1.1µg/dL) and Mg +2 ions (68.3± 8.6mg/dL) were significantly increased (p<0.01) compared with their concentrations in non-hemolysed samples. Alkaline phosphatase activity was inversely proportional with the increase in the hemoglobin concentrations in the hemolysed samples. A significant decrease (p<0.005) in the activity of ALP was observed after the addition of 68.3.0 mg/dl of magnesium chloride to group A. There was no significant decrease (p>0.1) in activity of ALP in the samples of group B. The findings of this study indicate that blood cell hemolysis reduces the activity of ALP which is directly proportional to the level of hemolysis. Released Mg +2 ions were found to inhibit ALP activity in the blood hemolysed samples.
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ABSTRACT: Alcoholic liver disease (ALD) is linked to binge drinking and cigarette smoking. Heavy chronic ± binge alcohol, or low-level exposures to dietary nitrosamines cause steatohepatitis with insulin resistance and oxidative stress in animal models. This study examines hepatotoxic effects of sub-mutagenic exposures to tobacco-specific nitrosamine (NNK) in relation to ALD. Long Evans rats were fed liquid diets containing 0 or 26% (caloric) ethanol (EtOH) for 8 weeks. In Weeks 3 through 8, rats were treated with NNK (2 mg/kg) or saline by i.p. injection, 3×/week, and in Weeks 7 and 8, EtOH-fed rats were binge-administered 2 g/kg EtOH 3×/week; controls were given saline. EtOH ± NNK caused steatohepatitis with necrosis, disruption of the hepatic cord architecture, ballooning degeneration, early fibrosis, mitochondrial cytopathy and ER disruption. Severity of lesions was highest in the EtOH+NNK group. EtOH and NNK inhibited insulin/IGF signaling through Akt and activated pro-inflammatory cytokines, while EtOH promoted lipid peroxidation, and NNK increased apoptosis. O(6)-methyl-Guanine adducts were only detected in NNK-exposed livers. Both alcohol and NNK exposures contribute to ALD pathogenesis, including insulin/IGF resistance and inflammation. The differential effects of EtOH and NNK on adduct formation are critical to ALD progression among alcoholics who smoke. © The Author 2015. Medical Council on Alcohol and Oxford University Press. All rights reserved.Alcohol and alcoholism (Oxford, Oxfordshire). Supplement 01/2015; 50(2). DOI:10.1093/alcalc/agu083