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Introduction: Our aim was to investigate the stability of clinically relevant analytes in pleural and peritoneal fluids stored in variable time periods and variable storage temperatures prior to analysis. Materials and methods: Baseline total proteins (TP), albumin (ALB), lactate dehydrogenase (LD), cholesterol (CHOL), triglycerides (TRIG), creatinine (CREA), urea, glucose and amylase (AMY) were measured using standard methods in residual samples from 29 pleural and 12 peritoneal fluids referred to our laboratory. Aliquots were stored for 6 hours at room temperature (RT); 3, 7, 14 and 30 days at - 20°C. At the end of each storage period, all analytes were re-measured. Deviations were calculated and compared to stability limits (SL). Results: Pleural fluid TP and CHOL did not differ in the observed storage periods (P = 0.265 and P = 0.170, respectively). Statistically significant differences were found for ALB, LD, TRIG, CREA, urea, glucose and AMY. Peritoneal fluid TP, ALB, TRIG, urea and AMY were not statistically different after storage, contrary to LD, CHOL, CREA and glucose. Deviations for TP, ALB, CHOL, TRIG, CREA, urea and AMY in all storage periods tested for both serous fluids were within the SL. Deviations exceeding SL were observed for LD and glucose when stored for 3 and 7 days at - 20°C, respectively. Conclusions: TP, ALB, CHOL, TRIG, CREA, urea and AMY are stable in serous samples stored up to 6 hours at RT and/or 30 days at - 20°C. Glucose is stable up to 6 hours at RT and 3 days at - 20°C. The stability of LD in is limited to 6 hours at RT.

Within the last several years, frequency of vitamin D testing has multiplied substantially all over the world, since it has been shown to have an important role in many diseases and conditions. Even though liquid chromatography - tandem mass spectrometry (LC-MS/MS) has been identified as “gold standard” method for vitamin D measurement, most laboratories still use immunochemistry methods. Besides analytical problems (hydrophobicity, low circulating concentrations, ability to bind to lipids, albumins and vitamin D binding protein, presence of multiple vitamin D metabolites and variable ratios of 25(OH)D2 and 25(OH)D3 in the blood), vitamin D shows great preanalytical variability, since its concentration is drastically influenced by seasonal changes, exposure to sun, type of clothes or sun block creams. Vitamin D is mostly measured in serum or plasma, but new studies are showing importance of measuring vitamin D in pleural effusions, breast milk, urine, synovial fluid and saliva. Besides the main role in calcium homeostasis and bone metabolism, many studies linked vitamin D deficiency with cancer, cardiovascular diseases, diabetes, fertility and many other conditions. However, even though initial observational studies indicated that supplementation with vitamin D might be beneficial in disease development and progression; first results of well-designed randomized controlled prospective studies did not find differences in frequency of cardiovascular events or invasive cancer between patients taking vitamin D supplementation compared to placebo. In the light of these recent findings, validity of excessive vitamin D testing remains an open question.

Background:The aim of our study was to perform verification of serum indices on three clinical chemistry platforms.Methods:This study was done on three analyzers: Abbott Architect c8000, Beckman Coulter AU5800 (BC) and Roche Cobas 6000 c501. The following analytical specifications were verified: precision (two patient samples), accuracy (sample with the highest concentration of interferent was serially diluted and measured values compared to theoretical values), comparability (120 patients samples) and cross reactivity (samples with increasing concentrations of interferent were divided in two aliquots and remaining interferents were added in each aliquot. Measurements were done before and after adding interferents).Results:Best results for precision were obtained for the H index (0.72%–2.08%). Accuracy for the H index was acceptable for Cobas and BC, while on Architect, deviations in the high concentration range were observed (y=0.02 [0.01–0.07]+1.07 [1.06–1.08]x). All three analyzers showed acceptable results in evaluating accuracy of L index and unacceptable results for I index. The H index was comparable between BC and both, Architect (Cohen’s κ [95% CI]=0.795 [0.692–0.898]) and Roche (Cohen’s κ [95% CI]=0.825 [0.729–0.922]), while Roche and Architect were not comparable. The I index was not comparable between all analyzer combinations, while the L index was only comparable between Abbott and BC. Cross reactivity analysis mostly showed that serum indices measurement is affected when a combination of interferences is present.Conclusions:There is heterogeneity between analyzers in the hemolysis, icteria, lipemia (HIL) quality performance. Verification of serum indices in routine work is necessary to establish analytical specifications.

Background The aim of this work was to determine the frequency of ascorbic acid-positive urine samples and to evaluate the potential risk on patient safety. Materials and methods A total of 27,856 samples submitted for urinalysis in previous year were retrospectively analysed from the laboratory information system. Urinalysis was done on Iris IQ200 analyzer using 11-parameter iChem Velocity test strips. Risk analysis combined the impact of potentially erroneous results (due to ascorbic acid interference) on patient safety (severity – S) with their frequency (occurrence – O). Potential risk was identified as high, intermediate and low. Results One thousand one hundred and ninety-nine (4.3%) and 1646 (5.9%) samples were mildly (20 mg/dL, 1+) and highly positive (40 mg/dL, 2+) for ascorbic acid, respectively. Severity (S1 to S5) was assigned to four possible errors: false-negative glucose (U-Glc: S2), blood (U-Hb: S5), nitrite (U-Nit: S4) and bilirubin (U-Bil: S1). Based on the frequency of errors, occurrence was categorized as: O2 for U-Glc, and O3 for U-Bil, U-Nit and U-Hb. The risk analysis matrix revealed that false-negative U-Hb was associated with potentially high risk, false-negative U-Nit with intermediate risk, while false-negative U-Glc and U-Bil were associated with low risk on patient safety. Conclusions Positive ascorbic acid can cause potentially high risk for patient safety by missing positive blood in urine.

Background Serum samples should be centrifuged for at least 10 min at 1300–2500 × g. Changed centrifugation conditions could compromise sample quality. The objective of this study was to compare the serum quality and turnaround time (TAT) using different centrifugation conditions. Methods The study was done in four different periods (A, B, C and D) at different conditions: for 10, 5 and 7 (A, B and C, respectively) at 2876 × g, and 7 (D) min at 4141 × g. Sample quality was assessed as the proportion of samples with: (a) aspiration errors, (b) H index >0.5 g/L and (c) suppressed reports of potassium (K) due to hemolysis. TAT was calculated for emergency samples. The proportions of samples (a), (b) and (c) were compared according to period A. Results The number of aspiration errors was significantly higher in samples centrifuged at 2876 × g for 5 min (p = 0.021) and remained unchanged when centrifuged for 7 min (p = 0.066 and 0.177, for periods C and D, respectively). In periods B, C and D, the proportion of samples with hemolysis was higher than that in period A (p-values 0.039, 0.009 and 0.042, respectively). TAT differed between all periods (p Conclusions The integrity of serum samples is changed with different centrifugation conditions than those recommended. Our study showed that shorter centrifugation at higher force (7 min at 4141 × g) significantly decreases TAT, with unchanged proportion of samples with aspiration errors.

Background Nowadays over-the-counter (OTC) drugs and dietary supplements are widely used. Their use can have a significant impact on the validity of laboratory results. The aim of this multicenter European study was to determine the frequency of consumption of various dietary products and OTC drugs among patients and explore their level of knowledge and awareness about the potential impact of various products on laboratory test results. Methods Eighteen European countries participated in this study. The survey was carried out anonymously on a subsequent series of outpatients (n=200) in each participating country. Included were patients who were referred to the laboratory for blood sampling and who voluntarily agreed to participate in the study. The survey included questions about the frequency of consumption of various products, awareness of the importance of informing physicians and laboratory staff about it and information about influence of preanalytical factors in general on laboratory test results. Results In total, 68% of patients were regularly taking at least one OTC drug or dietary supplement. The frequency of patients consuming at least one OTC drug or dietary supplement differed between countries (p=0.001). Vitamins (38%), minerals (34%), cranberry juice (20%), acetylsalicylic acid (ASA) (17%) and omega fatty acids (17%) were the most commonly used in our study. Conclusions The use of various OTC drugs and dietary supplements is highly prevalent in Europe and patients are often not willing to disclose this information to the laboratory staff and ordering physician. The education of both patients and healthcare staff is needed.

Objective: To determine whether K2-ethylenediaminetetraacetic acid (EDTA) and K3-EDTA Greiner tubes could be used interchangeably for glycosylated hemoglobin, type A1C (HbA1c) measurement via the Abbott Laboratories ARCHITECT chemiluminescent microparticle HbA1c assay on the ARCHITECT i 2000SR immunoanalyzer at our university hospital. Materials: We drew blood from a total of 45 outpatients into plastic Greiner Vacuette tubes, some of which were lined with K2-EDTA and others with K3-EDTA anticoagulant. Data are presented as median and interquartile range values. We used the Wilcoxon test and Passing-Bablok regression for tube comparison. Results: For K2-EDTA tubes median HbA1c concentration was 54 mmol/mol (41 to 71 mmol/mol) and for K3-EDTA tubes 56 mmol/mol (43 to 69 mmol/mol). There was no statistically significant difference between K2-EDTA and K3-EDTA (bias= -1.29 mmol/mol; P = 0.24). Passing-Bablok regression showed that there is no constant and proportional error: y = ‐0.23 (95% CI[-3.52 to 0.69]) + 1.00( 95% CI[0.98 to 1.06]) x . Conclusion: In this study, we provide evidence for the lack of any clinically and statistically significant bias between K2-EDTA and K3-EDTA HbA1c measurements. Thus, Greiner tubes lined with K2-EDTA and those lined with K3-EDTA can safely be used interchangeably to measure HbA1c via the Abbott Laboratories ARCHITECT assay. * Abbreviations : HbA1c : glycosylated hemoglobin, type A1C EDTA : ethylenediaminetetraacetic acid ICSH : International Council for Standardization in Hematology CLSI : Clinical and Laboratory Standards Institute CV : coefficient of variation Hb : hemoglobin CI : confidence interval

Endogenous interferences are an important source of biased laboratory results. Hemolysis, lipemia and icteria are the main source of endogenous interference in laboratory medicine. Accreditation according to ISO 15189 improves the overall quality of the laboratory procedures. The aim of our study was i) to assess the level of knowledge of Croatian medical biochemists about the proper detection and management of hemolysis, lipemia and icteria; and ii) to identify possible differences in the level of knowledge respective to the laboratory accreditation status.An on-line self-report survey was carried out by the Working Group for Preanalytical Phase of the Croatian Society of Medical Biochemistry and Laboratory Medicine during April to May 2015. Survey included 14 statements (Q1–Q14) about procedures for samples with interferences and participants were asked to assess the degree of agreement with the statement using a 4-point Likert scale.The lowest level of knowledge was observed for statements Q10 (dealing with icteric sample; 40.9% participants agreed with the correct procedure), Q12 (allowable error for interference; 47.2%) and Q11 (dealing with lipemic sample; 60.1%). Almost all participants (97.4%) agreed that laboratories in Croatia should have a harmonized protocol for management of samples with interferences. Participants from accredited laboratories showed higher knowledge of hemolysis detection (p=0.031), rejection of hemolyzed sample (pCroatian laboratories have a good knowledge of the proper detection and management of hemolyzed, icteric and lipemic samples. Accreditation is associated with higher knowledge about management of samples with interferences.