Hemolyzed Samples Research Articles

Effect of Hemolysis on Serum Markers of Liver Function

Introduction Samples received in the laboratory are often rejected because of visible hemolysis. Every laboratory has protocols on whether to accept or reject hemolyzed samples based on tests ordered. The objective of the study was to observe and analyze the variations seen in association with hemolyzed samples in liver function test parameters and electrolytes measurement. MethodsThis cross-sectional observational study was carried out in 88 laboratory samples in the Clinical Biochemistry Laboratory. Hemolysis was induced by mechanical mixing. This study compared reported values of bilirubin, protein, albumin, aminotransferases (ALT and AST), alkaline phosphatase (ALP), gamma glutamyl transpeptidase (GGT), sodium and potassium between hemolyzed and non-hemolyzed samples. ResultsMean values of total bilirubin (TB) and direct bilirubin (TB), ALT, ALP, GGT and sodium (Na) were found to be significantly decreased post hemolysis (p<=0.001) with mean variances (σ2) at 0.30 (TB), 0.38 (DB), 0.56 (ALT), 0.18 (ALP), 0.01 (GGT) and 0.001 (Na). In contrast, mean values of AST (σ2= -0.9, p<0.001) and potassium (σ2 = -0.3, p<0.001) were found to be significantly elevated post hemolysis. Total protein and albumin were also found to be elevated in hemolyzed samples but the average variance was not statistically significant. Degree of hemolysis had highly varying effects in some parameters like AST and total protein, but less varying and more consistent effects on other measured parameters. ConclusionFree hemoglobin estimation should be considered before rejecting a clinical sample because of hemolysis. Mathematical equations cannot be expected to reproduce corrected values of the analytes in question because of the large variances, so these tests need to be repeated on non-hemolyzed samples for the analytes that are affected.Keywords

Potential medical impact of unrecognized invitro hypokalemia due to hemolysis: a case series.

The destruction of red cells during blood collection or with the processing of the sample continues to occur at a high rate, especially among emergency department (ED) patients. This can produce pre-analytical laboratory errors, particularly for potassium. We determined the incidence of hemolyzed samples and discuss the potential medical impact for hypokalemic patients who potassium level is artificially normal (pseudoeukalemia). Potassium results were obtained for a 6-month period. Using a measured hemolysis index (HI), hemolysis was present in 3.1 % for all potassium ordered (n=94,783) and 7.5 % for ED orders (n=22,770). Most of these samples were reported as having high normal result or were hyperkalemic. There were 22 hemolytic samples with a potassium of <3.5 mmol/L, and 57 hemolytic samples with a potassium in lower limit of normal (3.5-3.8 mmol/L). From this group, we examined the medical histories of 8 selected patients whose initially normal potassium levels were subsequently confirmed to have a potassium values that were below, at, or just above the lower limit of normal due to hemolysis. The primary complaint for these patients were: necrotizing soft tissue infection, pancreatitis, volume overload from heart failure with reduced ejection fraction, hypertension treated with hydrochlorothiazide, and presence of a short bowel syndrome. A subsequent non-hemolyzed sample was collected demonstrating hypokalemia in all of these patients. Within these cases, there was a potential for harm had hemolysis detection not been performed. We demonstrate the medical importance of detecting hemolysis for patients who have pseudoeukalemia. This is relevant because the HI cannot be obtained when electrolytes are tested using whole blood samples, and a normal potassium may lead to inappropriate patient management.

Ultrabright NIR AIEgen nanoparticles‐enhanced lateral flow immunoassay platform for accurate diagnostics of complex samples

AbstractAccurate and sensitive near‐infrared (NIR) luminescent lateral flow immunoassay (LFIA) has attracted considerable attention in the field of point‐of‐care testing (POCT). However, the detection accuracy and sensitivity are often compromised by the low fluorescence quantum efficiency (&lt;10%) of the NIR fluorescent probe. Herein, ultrabright NIR AIEgen nanoparticles (PS@AIE830NPs) composed of polystyrene (PS) nanoparticles and NIR aggregation‐induced emission luminogen (AIEgen) with the maximum emission at 830 nm (AIE830) is reported, and its potential to promote an accurate and sensitive detection of complex samples by LFIA is described. The relative quantum yield (QY) of the PS@AIE830NPs was 14.76%, which was superior to that of the polymer embedding method and indocyanine green (ICG)‐based NIR nanoparticles. The PS@AIE830NPs immunolabeled‐LFIA combined with laboratory‐built NIR‐LFIA portable quantitative instruments (detected light range &gt; 800 nm) completely eliminated background interference and allowed highly accurate and sensitive detection without any pre‐treatment steps. The limits of detection (LODs) for aflatoxin B1 (AFB1) in soy sauce, alpha hemolysin (Hla) of Staphylococcus aureus biomarker in joint fluid, and C‐reactive protein (CRP) in human haemolysed samples were 0.01 ng mL−1, 0.02 µg mL−1, and 0.156 mg L−1, respectively, commensurating with those of the corresponding gold standard assays and covering the detection range of interests. It is anticipated that the ultrabright NIR AIEgen nanoparticles will serve as a universally applicable signal probe for NIR‐LFIA diagnostics, promising to expand the range of applications for quantitative detection of complex samples.

Can Patients with Electrolyte Disturbances Be Safely and Effectively Treated in a Hospital-at-Home, Telemedicine-Controlled Environment? A Retrospective Analysis of 267 Patients.

Morbidities indicated for hospital-at-home (HAH) treatment include infectious diseases and exacerbations of chronic conditions. Electrolyte disturbances are not included per se. However, their rate is high. We aimed to describe our experience via the monitoring and treatment of such patients. This was a retrospective analysis of patients in the setting of telemedicine-controlled HAH treatment. We collected data from the electronic medical records of patients who presented electrolyte disturbances. For 14 months, we treated 267 patients in total in HAH settings, with a mean age of 72.2 + 16.4, 44.2% for males. In total, 261 (97.75%) patients were flagged with electrolyte disturbances, of whom 149 had true electrolyte disturbances. Furthermore, 67 cases (44.96%) had hyponatremia, 9 (6.04%) had hypernatremia after correction for hyperglycemia, 20 (13.42%) had hypokalemia and 27 (18.12%) had hyperkalemia after the exclusion of hemolytic samples. Ten (6.09%) patients had hypocalcemia and two (1.34%) had hypercalcemia corrected to albumin levels. Thirteen (8.72%) patients had hypomagnesemia and one (0.67%) had hypermagnesemia. Patients with electrolyte disturbances suffered from more chronic kidney disease (24.2% vs. 12.2%; p = 0.03) and malignancy (6.3% vs. 0.6%; p = 0.006), and were more often treated with diuretics (12.6% vs. 4.1%; p = 0.016). No patient died or suffered from clinically significant cardiac arrhythmias. The extent of electrolyte disturbances amongst HAH treatment patients is high. The monitoring and treatment of such patients can be conducted safely in this setting.

Red Blood Cell Storage with Xenon: Safe or Disruption?

Xenon, an inert gas commonly used in medicine, has been considered as a potential option for prolonged preservation of donor packed red blood cells (pRBCs) under hypoxic conditions. This study aimed to investigate how xenon affects erythrocyte parameters under prolonged storage. In vitro model experiments were performed using two methods to create hypoxic conditions. In the first method, xenon was introduced into bags of pRBCs which were then stored for 42 days, while in the second method, xenon was added to samples in glass tubes. The results of our experiment showed that the presence of xenon resulted in notable alterations in erythrocyte morphology, similar to those observed under standard storage conditions. For pRBC bags, hemolysis during storage with xenon exceeded the acceptable limit by a factor of six, whereas the closed-glass-tube experiment showed minimal hemolysis in samples exposed to xenon. Notably, the production of deoxyhemoglobin was specific to xenon exposure in both cell suspension and hemolysate. However, this study did not provide evidence for the purported protective properties of xenon.

Hemolysis in blood samples

Hemolysis is one of the interfering factors that can limit the results of various biochemical tests, and this study aimed to assess the interference of hemolysis in the reliability of biochemical tests conducted in clinical-surgical procedures in the veterinary hospital routine. The methodology was retrospective, based on the analysis of records of animals treated at the Veterinary Hospital Unit (SUHVU) of the Federal University of Fronteira, Campus Realeza (Paraná) from January of 2020 to December of 2022. The tests conducted in the Veterinary Clinical Analysis laboratory included serum determination of total proteins, albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), creatinine and urea. Based on the biochemical results, animals were divided into three groups according to the degree of hemolysis observed in the serum: mild hemolysis (HD); moderate hemolysis (HM) and intense hemolysis (HI). The control group (GC) was determined based on reference values specific to the species. Data were tabulated in spreadsheets and analyzed using Pearson's correlation test (p). Mild hemolysis was predominant and, in dogs, did not affect the analyzed biochemical values in comparison to the reference averages for the species, regardless of the degree. Isolated, most measurements increased. In cats, changes were more significant concerning the group average, with ALP showing the greatest variation, not consistent with the elevation of hemolysis. None of the parameters had a strong correlation of interference between hemolyses, only moderate negative and positive correlations in ALP and urea.

Active targeted delivery of theranostic thermo/pH dual-responsive magnetic Janus nanoparticles functionalized with folic acid/fluorescein ligands for enhanced DOX combination therapy of rat glioblastoma.

Doxorubicin (DOX), a chemotherapy drug, has demonstrated limited efficacy against glioblastoma, an aggressive brain tumor with resistance attributed to the blood-brain barrier (BBB). This study aims to overcome this challenge by proposing the targeted delivery of magnetic Janus nanoparticles (MJNPs) functionalized with folic acid ligands, fluorescent dye, and doxorubicin (DOX/MJNPs-FLA). The properties of these nanoparticles were comprehensively evaluated using bio-physiochemical techniques such as Fourier transform infrared (FTIR) spectroscopy, dynamic light scattering (DLS), zeta potential analysis, high-resolution transmission electron microscopy (HR-TEM), vibrating sample magnetometry (VSM), fluorescence microscopy, MTT assay, hemolysis assay, and liver enzyme level evaluation. Dual-controlled DOX release was investigated under different pH and temperature conditions. Additionally, the impact of DOX/MJNPs-FLA on apoptosis induction in tumor cells, body weight, and survival time of cancerous animals was assessed. The targeted delivery system was assessed using C6 and OLN-93 cell lines as representatives of cancerous and healthy cell lines, respectively, alongside Wistar rat tumor-bearing models. Results from Prussian blue staining and confocal microscopy tests demonstrated the effective targeted internalization of MJNPs-FLA by glioblastoma cells. Additionally, we investigated the biodistribution of the nanoparticles utilizing fluorescence imaging techniques. This enabled us to track the distribution pattern of MJNPs-FLA in vivo, shedding light on their movement and accumulation within the biological system. Furthermore, the combination of chemotherapy and magnetic hyperthermia exhibited enhanced efficacy in inducing apoptosis, as evidenced by the increase of the pro-apoptotic Bax gene and a decrease in the anti-apoptotic Bcl-2 gene. Remarkably, this combination treatment did not cause any hepatotoxicity. This study highlights the potential of DOX/MJNPs-FLA as carriers for therapeutic and diagnostic agents in the context of theranostic applications for the treatment of brain malignancies. Additionally, it demonstrates the promising performance of DOX/MJNPs-FLA in combination treatment through passive and active targeting.

Multicenter comparison of analytical interferences of 25-OH vitamin D immunoassay and mass spectrometry methods by endogenous interferents and cross-reactivity with 3-epi-25-OH-vitamin D3

BackgroundVitamin D (vit-D) deficiency is highly prevalent in the Korean population, highlighting the need for accurate measurements. In this study, the interferences by endogenous and cross-reactive substances were compared between routine vit-D immunoassays and mass spectrometry (MS) methods. MethodsTwo MS methods and 4 immunoassays from different manufacturers (Abbott, Beckman Coulter, Roche, Siemens) were compared. Residual samples that were icteric, lipemic, hemolyzed, high in rheumatoid factor, from myeloma patients, or patients undergoing hemodialysis were collected. Also, 4 levels of National Institute of Standards and Technology (NIST) Standard Reference Material 972a, and 12 samples serially spiked with 3-epi-25-OH-D3 were prepared. ResultsSignificant interferences were observed in hemolytic (Roche), icteric (Beckman and Siemens) and lipemic samples (all 4 immunoassays). Level 4 NIST material and 3-epi-25-OH-D3-spiked samples induced significant cross-reactivity, yielding higher total vit-D measurements in non-epimer-separating MS methods, and both the Beckman and Roche immunoassays. ConclusionMost observed interferences were consistent with manufacturers’ claims, but overall improvement of immunoassay bias limits is required. Awareness of potential interference is important to increase the accuracy of vit-D measurements. Moreover, care is due when interpreting vit-D results of newborns, infants and less commonly, pregnant women, who are known to have physiologically high levels of the highly cross-reactive 3-epi-25-OH-D3.

Abstract C103: Preclinical and clinical safety and tolerability evaluation of NBF-006, a novel siRNA inhibitor of glutathione-s-transferase P (GSTP) encapsulated in a lipid nanoparticle (LNP) for treatment of advanced non-small cell lung cancer (NSCLC)

Abstract Glutathione S-transferase Pi (GSTP) has an important role in detoxification and anti-oxidative damage responses. Additionally, GSTP has a chaperone function that regulates key oncogenic pathways such as the KRAS and JNK pathways (E Laborde, 2010). Since alternative or redundant pathways could compensate for the incomplete inhibition of a single kinase, targeting multiple pathways could lead to more effective therapy. NBF-006 is a novel drug product containing a GSTP siRNA encapsulated within a proprietary lipid nanoparticle (LNP). It has been designed to deliver siRNA to tumors of the lung and common sites of metastatic spread (liver and marrow). As GSTP downregulation leads to inhibition of KRAS signaling through modulation of both MAPK and PI3K pathways, NBF-006 is being developed to treat KRAS mutant tumors. Preclinical studies have demonstrated activity across KRAS mutations, as well as efficacy in orthotopic non-small cell lung cancer (NSCLC) models. Immune toxicities have been recognized as safety concerns associated with LNP therapeutics. The safety assessment of NBF-006 was conducted both preclinically and clinically. In vitro preclinical toxicity studies included hemolysis in hRBC samples, cytokine release in hPBMCs, and TLR activation in human THP1 cells. GLP-compliant general toxicity studies were conducted in both mice and cynomolgus monkeys, IV administration, once a week, for 5 weeks. Complement splits were measured in the monkey study to assess any immune-related responses. Safety pharmacology parameters were also conducted in monkeys. The mouse was slightly more sensitive than monkeys to NBF-006, and the highest non-seriously toxic dose level in mice was 20 mg/kg/week, which yielded a good safety margin to proceed for a Phase I clinical trial. The highest dose level tested in monkeys was of 18 mg/kg/week and was well tolerated by the monkeys. Immunological assessments were conducted during Phase 1 dose escalation and expansion in patients with NSCLC, pancreatic cancer, or colorectal cancer. NBF-006 was administered by IV infusion over 70 minutes, once a week, for 4 weeks followed by a 2-week rest period. Clinical symptoms and reactions were monitored, and lab tests were conducted according to the protocol, including CBC, complement split products (Bb, C3a, C5a, and CH50), cytokines (IL-6, IL-1β, IFNg, and TNFα), and anti-drug antibodies against the LNPs. Dose-dependent increases in C3a and Bb of 1.2 and 1.6 mg/kg were observed following NBF-006 administration at doses ranging from 0.15 to 1.6 mg/kg. The increases were transient at either 10 minutes or 1 hour after the end of the infusion. Due to lack of corresponding increases in C5a or decreases in CH50, transient elevations of C3a and Bb were not considered clinically significant. In summary, NBF-006 was well tolerated in both animal species and patients: 20 mg/kg/week was the non-seriously toxic dose level in mice; the highest dose level of 18 mg/kg/week tested in monkeys was the NOAEL; and 1.6 mg/kg/week was the highest dose level evaluated in humans. Citation Format: Jian Liu, Cima Cina, Douglas Kornbrust, Sonya Zabludoff. Preclinical and clinical safety and tolerability evaluation of NBF-006, a novel siRNA inhibitor of glutathione-s-transferase P (GSTP) encapsulated in a lipid nanoparticle (LNP) for treatment of advanced non-small cell lung cancer (NSCLC) [abstract]. In: Proceedings of the AACR-NCI-EORTC Virtual International Conference on Molecular Targets and Cancer Therapeutics; 2023 Oct 11-15; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2023;22(12 Suppl):Abstract nr C103.

Errors in measurement of glomerular filtration rate using the slope-intercept technique and their identification.

GFR measured from plasma sampling may be expressed as slope-intercept GFR (SI-GFR) and scaled to body surface area (mGFR/BSA) or as GFR per unit extracellular fluid volume (mGFR/ECV), which is based only on half-time. Measurement errors comprise 3 categories. Pre-injection error arises from error in administered marker and is suspected when mGFR/BSA and mGFR/ECV disagree. Injection errors include 'tissued' injections. Post-injection errors include inaccurate sample timing, inaccurate pipetting, sample haemolysis and sampling through long IV lines through which marker was administered. The aim of the study was to evaluate the impact of errors on mGFR. We compared mGFR/BSA with mGFR/ECV in 898 patients undergoing routine investigation. To investigate post-injection error, we took two further patient datasets with r values (correlation coefficient of the 3-sample fit) of 1.0 and introduced errors, in isolation, into each of the 3 recorded sample values, as follows: pipetting (volume) errors of -20%, -10%, -5%, 5%, 10% and 20%, and timing errors of -15 min, -10 min, -5 min, 5 min, 10 min and 15 min. The correlation between mGFR/BSA and mGFR/ECV was close and independent of r. Post-injection error depended on the time of the sample in which it occurred. r correlated poorly with error magnitude for both volume and timing errors. When a 'rogue' sample is suspected its error needed to be substantial for it to be identified by single sample estimates applied to the other samples. SI-GFR is resistant to post-injection timing and volume errors but not to pre-injection error.

Rapid serum tubes reduce transport hemolysis and false positive rates for high-sensitivity troponin T

IntroductionHemolysis in the emergency department (ED) can significantly delay results and appropriate action. We evaluated the main sources of hemolysis during sample collection, and to evaluate the use of rapid serum tubes (RST) as a transport hemolysis-mitigating measure for high-sensitivity troponin T (hs-cTnT) testing. MethodsWe examined the effect of tube type, tube fill, types of sample draw and collection methods on hemolysis and hs-cTnT in samples (n = 158) from ED patients. We also compared hs-cTnT values in paired RST and plasma separate tube (PST) samples that were hemolysis-free. ResultsThe primary source of hemolysis in samples collected in the ED was underfilling tubes. In both tube types, PST and RST, filled tubes showed a median reduction in hemolysis of 69.1 % (p < 0.0001). Blood collected in RST also experienced less hemolysis compared to PST. In hemolysis-free samples, false positive results in PST were noted in patients with hs-cTnT values < 50 ng/l. ConclusionWe suggest that proper tube filling during sample collection and use of RST tubes can significantly reduce the effects of hemolysis. In addition, laboratories should be aware that PST tubes have a non-trivial rate of false positives when hs-cTnT < 50 ng/l.

DETERMINE PRE-ANALYTICAL ERRORS ' PERCENTAGE IN CLINICAL BIOCHEMISTRY LABS AND DEVELOP REMEDIAL PROCEDURES FOR THEIR PREVENTION

Diagnostic inaccuracies pose a formidable issue in Laboratory Medicine, necessitating immediate initiatives to expand understanding and minimize these discrepancies. Our study delves into the nuances of errors associated with laboratory testing processes, from the test request to result interpretation, underlining the imperative for reducing such errors. Our investigation seeks to identify the incidence of errors in a round-the-clock lab setting over four months, covering pre-analytical, analytical, and post-analytical phases.Over a span period from Jan 2021 to Jan 2022, a broad spectrum of errors within pre-analytical, analytical, and post-analytical phases was methodically logged using a bespoke proforma. The collected data was systematically analyzed utilizing SPSS version 22.All venous and arterial blood samples were processed in the 24-hour lab throughout this period. From the 185,012 samples received, 2,450 were deemed unsuitable for testing, accounting for a 1.32% rejection rate. These samples were disqualified due to an array of pre-analytical errors, including misidentification (0.07%), incorrect tube usage (0.11%), missing samples (0.07%), drawing from an intravenous site (0.10%), inadequate sample volume (0.51%), improper timing of sample collection (0.10%), hemolysed samples (0.31%), and lipemic samples (0.11%).Utilizing an agreed list of evidence-based Quality Indicators, compliant with the current International Standard (ISO 15189:2012), for incorporation into accreditation programs of clinical laboratories stands as a potent approach to augment quality, mitigate error risks, and bolster patient safety.

A-034 Multicenter Comparison of Analytical Interference of Vitamin D Immunoassay and Mass Spectrometry Methods by Endogenous Interferents and Cross-reactivity with 3-epi-25-OH-Vitamin D3

Abstract Background Vitamin D plays an important role in calcium homeostasis and bone metabolism and is known to increase calcium absorption from the intestinal tract and decrease calcium excretion from the kidneys. Vitamin D deficiency is prevalent in a substantial and continuously increasing proportion of the Korean population, thus highlighting the need for accurate vitamin D measurements. The current problems of vitamin D testing include significant inter-method and inter-laboratory differences in measurements, and interferences by both intrinsic and cross-reactive substances. In this study, the interfering effects by such intrinsic and cross-reactive substances were compared between several vitamin D immunoassays and mass spectrometry methods routinely used in Korea. Methods Two different mass spectrometry methods and 4 vitamin D immunoassays with different manufacturers (Abbott, Beckman Coulter, Roche, Siemens), from 5 different institutes were compared. Residual patient samples were utilized to collect either hemolyzed samples, icteric samples (total bilirubin &amp;gt;20 mg/dL), lipemic samples (triglyceride &amp;gt;500 mg/dL), of samples with rheumatoid factor &amp;gt;200 IU/mL, samples from myeloma patients (paraprotein &amp;gt;1.1 g/dL upon protein electrophoresis) or from patients undergoing hemodialysis. Four levels of National Institute of Standards and Technology (NIST) Standard Reference Material (SRM) 972 were prepared, with each level having different compositions and concentrations of vitamin D. Also, samples spiked with different levels of 3-epi-25(OH)D3 were prepared for interference analysis. The 10% bias limits for concentrations of intrinsic interferents and cross-reactivity percentages, provided by the respective immunoassay manufacturers, were referred to during data interpretation. The collection of intrinsic interferent samples, NIST material, and epimer spiked sample sets were delivered to each institute and measured on the same date. Results Consistently significant interference in vitamin D measurements was observed in hemolytic samples (Roche assay), icteric samples (Beckman and Siemens assays) and lipemic samples (all 4 immunoassays). General positive biases were also observed in myeloma samples (Abbott assay), hemodialysis samples (Siemens assay), and general negative biases were observed in high rheumatoid factor (Roche assay) and hemodialysis samples (Roche assay). Both the level 4 NIST material, which had a high concentration of 3-epi-25(OH)D3, and samples spiked with 3-epi-25(OH)D3, induced significant interference, yielding higher total vitamin D measurements in non-epimer separating MS methods, and the Beckman and Roche immunoassays. Conclusion In this study, we were able to observe interferences due to intrinsic substances and cross-reactive substances in common vitamin D tests used in Korea. Clinical awareness of possible causes of interference is important to increase the accuracy of vitamin D measurements. Moreover, due to the high cross-reactivity of 3-epi-25(OH)D3 with common vitamin D assays, particular care is due when interpreting vitamin D results of newborns, infants, and less commonly, pregnant women, who are known to have physiologically high levels of 3-epi-25(OH)D3.

A-382 Evaluation of the Labcorp TrueSpinTM for Decentralized Blood Sample Centrifugation

Abstract Background The increased demand for decentralized blood sample collection presents numerous operational challenges for diagnostics providers. Sample degradation including sample hemolysis due to time, temperature, and handling between collection and laboratory analysis leads to limited test menus and unreliable results. Here we evaluate the light-weight, portable Labcorp TrueSpin™ for rapid point-of-care blood separation using commercially available microvolume blood collection tubes. The TrueSpin is a class I FDA registered device designed for layperson use. The centrifuge runs on AA batteries and separates a blood sample in five minutes. Methods Here we show results of studies evaluating sample quality and analyte stability in serum samples collected into gel microtubes and processed using the TrueSpin. Hemolysis and residual red blood cell (RBC) concentration are compared between samples separated using the TrueSpin and conventional centrifugation. Additionally, we evaluate serum-based chemistry analyte stability in both separated and unseparated samples stored for three days refrigerated and unrefrigerated. Results No significant difference was seen in hemolysis or residual RBC concentration in serum samples prepared by TrueSpin compared to the reference centrifuge method. Furthermore, we show that many common serum-based chemistry analytes have limited (&amp;lt;1 day) stability if uncentrifuged, but improve to ≥3-day stability following TrueSpin separation and refrigerated or room temperature storage (see Table 1). Conclusions These findings suggest that the TrueSpin is a simple and effective solution for remote sample separation and may enable broader test menus and increased test result reliability for decentralized sample collection pursuits.

Evaluation of a Compact, Portable Centrifuge for Separating Microvolume Blood Samples at the Point of Collection.

The increased demand for decentralized blood sample collection presents numerous operational challenges for diagnostics providers. Sample degradation including sample hemolysis due to time, temperature, and handling between collection and laboratory analysis leads to limited test menus and unreliable results. Here we introduce the lightweight, portable Labcorp TrueSpin™ for rapid point-of-care blood separation using commercially available microvolume blood collection tubes. The TrueSpin is a class I FDA-registered device designed for untrained users. The centrifuge runs on AA batteries and separates a blood sample in 5 minutes. Here we describe a series of studies evaluating sample quality and analyte stability in serum samples collected into gel microtubes and processed using the TrueSpin. Hemolysis, residual red blood cell concentration, sample volume, and serum-based chemistry analyte stability were evaluated. No significant difference was seen in hemolysis or residual red blood cell concentration in serum samples prepared by TrueSpin compared to the reference method. Additionally, capillary and venous blood samples separated using the TrueSpin and exposed to International Safe Transit Association 3A-simulated shipping conditions were shown to yield acceptable sample volume and quality for laboratory analysis. Finally, we show that many common serum-based chemistry analytes have limited (< 1 day) stability if uncentrifuged but improve to ≥ 3-day stability following TrueSpin separation and refrigerated or room temperature storage. These findings suggest that the TrueSpin is a simple and effective solution for remote sample separation and may enable broader test menus and increased test result reliability for decentralized sample collection pursuits.

Confounders for prognostic accuracy of neuron-specific enolase after cardiac arrest: A retrospective cohort study

AimTo evaluate neuron-specific enolase (NSE) thresholds for prediction of neurological outcome after cardiac arrest and to analyze the influence of hemolysis and confounders. MethodsRetrospective analysis from a cardiac arrest registry. Determination of NSE serum concentration and hemolysis-index (h-index) 48–96 hours after cardiac arrest. Evaluation of neurological outcome using the Cerebral Performance Category score (CPC) at hospital discharge. Separate analyses considering CPC 1–3 and CPC 1–2 as good neurological outcome. Analysis of specificity and sensitivity for poor and good neurological outcome prediction with and without exclusion of hemolytic samples (h-index larger than 50). ResultsAmong 356 survivors three days after cardiac arrest, hemolysis was detected in 28 samples (7.9%). At a threshold of 60 µg/L, NSE predicted poor neurological outcome (CPC 4–5) in all samples with a specificity of 92% (86–95%) and sensitivity of 73% (66–79%). In non-hemolytic samples, specificity was 94% (89–97%) and sensitivity 70% (62–76%). At a threshold of 100 µg/L, specificity was 98% (95–100%, all samples) and 99% (95–100%, non-hemolytic samples), and sensitivity 58% (51–65%) and 55% (47–63%), respectively. Possible confounders for elevated NSE in patients with good neurological outcome were ECMO, malignancies, blood transfusions and acute brain diseases. Nine patients with NSE below 17 µg/L had CPC 5, all had plausible death causes other than hypoxic-ischemic encephalopathy. ConclusionsNSE concentrations higher than 100 µg/L predicted poor neurological outcome with high specificity. An NSE less than 17 µg/L indicated absence of severe hypoxic-ischemic encephalopathy. Hemolysis and other confounders need to be considered. Institutional protocol numberThe local ethics committee (board name: Ethikkommission der Charité) approved this study by the number: EA2/066/23, approval date: 28th June 2023, study title “‘ROSC’ – Resuscitation Outcome Study.”

FACTORS OF INTERFERENCE IN LABORATORY INVESTIGATIONS: HEMOLYSIS, ICTERUS, LIPEMIA

The accuracy of laboratory investigation results depends on various factors, including interfer- ence factors. Additionally, it is worth noting that a significant number of samples are photometrical- ly measured in the wavelength range of 350 to 600 nm, which corresponds to the absorption peak of many interferents.&#x0D; In laboratories where the process of sample rejection is established based on serum indices (color scale) and specific cutoff points for samples showing hemolysis, icterus, and lipemia, the issuance of falsely elevated/decreased investigation results to the clinic is avoided. The declared criteria for sample rejection should comply with the recommendations of CLSI CA-56A and CLSI EP 07 standards.&#x0D; The majority of errors in the preanalytical stage are caused by hemolysis. All hemolyzed sam- ples with a concentration of free hemoglobin equal to or greater than 10.0 g/L should be rejected, and the laboratory report should indicate the reason for rejection as "Hemolyzed sample. Resampling is required." Each laboratory establishes its own cutoff point for free hemoglobin, bili- rubin and triglicerol concentration in the sample. This is because an increase in hemolysis index in serum/plasma leads to falsely increased levels of certain analytes (such as inorganic phosphorus, potassium, folic acid, magnesium, iron, urea, LDH, AST, ALT). Conversely, the concentration of albumin, bilirubin, glucose, sodium, and chloride in such samples is falsely decreased. The presence of icterus in samples is always indicative of a pathological process. The elevation of bilirubin con- centration in the sample will be reflected in the increase/decrease of analytes such as creatinine, total protein, triglycerides, cholesterol, phosphorus, magnesium, urea, hormones (free triiodothyro-nine, testosterone, estradiol), and others. An increase in triglyceride and lipoprotein concentration in the sample significantly affects the levels of analytes such as phosphorus, creatinine, total protein and calcium.&#x0D; The presence of hemolysis, icterus, and lipemia in samples affects the measurement of various analytes and, consequently, the results of laboratory investigations and patient safety. Laboratories should have a developed procedure for systematic identification of interfering agents and decision- making regarding the handling of such samples to ensure the reliability of obtained data.

Increased preanalytical errors during Covid-19 Pandemic

The clinical laboratories worldwide have faced numerous obstacles due to the COVID-19 pandemic. In the midst of this outbreak, mistakes that occur during the preanalytical phase of sample collection, transportation, and processing can result in unfavorable clinical outcomes. Therefore, the aim of this investigation was to ascertain and contrast the rate at which blood specimens are rejected by a clinical laboratory and to analyze and compare the types of preanalytical errors during the pre-pandemic and pandemic periods. The present retrospective study was done in Chhatrapati Shivaji Subharti Hospital; India. Information was gathered from two distinct time periods - the pre-pandemic phase, which spanned from December 30th, 2019 to March 22, 2020, and the pandemic phase, which ran from March 23th to November 30, 2020. The rate of rejection for blood samples was established by determining the percentage of blood collection tubes containing preanalytical mistakes out of the total number of tubes received. A total of 102,000 blood samples were examined, out of which 40,800 (40%) were collected during the pandemic. The pandemic phase witnessed a significantly higher rate of rejection of blood samples (4%) as compared to the pre-pandemic phase (1%) (P &amp;#60; 0.001). In both phases, the most common preanalytical error was the clotting of samples. There was a significant increase in improperly labeled samples during the pandemic, while there was a notable decrease in hemolyzed samples. Given the prevailing situation of the Corona pandemic, altered logistics have led to a significant surge in preanalytical errors and consequent rejection of blood specimens in clinical laboratories. Therefore, it is imperative to take corrective measures at different levels to minimize preanalytical errors, which would ultimately enhance patient care and optimize the utilization of resources.

Hemolyzed Laboratory Specimens in the Emergency Department: An Underappreciated, but Frequent Problem

IntroductionHemolysis of blood samples from emergency department (ED) patients leads to delays in treatment and disposition. The aim of this study is to determine the frequency of hemolysis and variables predictive of hemolysis. MethodsThis observational cohort study was conducted among three institutions: academic tertiary care center and two suburban community EDs, with an annual census of over 270,000 ED visits. Data were obtained from the electronic health record. Adults requiring laboratory analysis with at least one peripheral intravenous catheter (PIVC) inserted within the ED were eligible. Primary outcome was hemolysis of lab samples and secondary outcomes included variables related to PIVC failure. ResultsBetween January 8, 2021 and May 9, 2022, 141,609 patient encounters met inclusion criteria. The average age was 55.5 and 57.5% of patients were female. Hemolysis occurred in 24,359 (17.2%) samples. In a multivariate analysis, when compared to 20-gauge catheters, smaller 22-gauge catheters had an increased odds of hemolysis (OR 1.78, 95% confidence interval (CI) 1.65-1.91; P < .001), while larger 18-gauge catheters had a lower odds of hemolysis (OR 0.94; 95% CI 0.90-0.98; P = .0046). Additionally, when compared to antecubital placement, hand/wrist placement demonstrated increased odds of hemolysis (OR 2.06; 95% CI 1.97-2.15; P < .001). Finally, hemolysis was associated with a higher rate of PIVC failure (OR 1.06; 95%CI 1.00-1.13; P = 0.043). DiscussionThis large observational analysis demonstrates that lab hemolysis of is a frequent occurrence among ED patients. Given the added risk of hemolysis with certain placement variables, clinicians should consider catheter gauge/placement location to avoid hemolysis that may result in patient care delays and prolonged hospital stays.

#5228 ASSESSING ACCURACY OF POINT-OF-CARE CREATININE AND POTASSIUM TESTING IN A MULTI-ETHNIC SOUTH LONDON POPULATION

Abstract Background and Aims Point-of-care (POC) testing allows rapid analysis of blood samples without delay of laboratory testing. For patients with chronic kidney disease (CKD), this allows for instant identification of changes in kidney function, such as acute kidney injury or hyperkalaemia, and assists in timely decision-making in outpatient or community settings (e.g. commencing or up-titrating medications dependent on renal function). It also provides an opportunity for early recognition of CKD in high-risk populations by providing screening in more convenient and trusted settings. To achieve these aims, it essential to confirm that POC systems can produce accurate results in real world settings with diverse patient populations. This pilot validation study assessed accuracy of the Siemens Epoc Blood Analysis System venous creatinine and potassium compared with laboratory assays in a diverse UK renal patient population. Method Venous blood samples from 57 patients, aged ≥ 18 years old and receiving care at a London teaching hospital, were analysed using the Siemens Epoc Blood Analysis System. POC-Creatinine (POC-Cr) and Potassium (POC-K) measurements were compared with laboratory (IDMS-traceable Siemens enzymatic and Roche) assays. Demographics (age, ethnicity and sex) were recorded for each patient. Passing-Bablock regression analysis were used to compare the results using both methods. Comparison between testing methods was assessed using a Bland-Altman plot. Limits of agreement were compared with CLIA criteria for acceptable performance for creatinine and potassium. Results The mean age was 55.6 ± 15.3 years old and 64.9% were male (n = 37). The majority of patients were of White (N = 23; 40.4%), Black (N = 21; 36.8%) or South Asian (N = 9; 15.8%) ethnicity respectively. Median POC-Cr was 216 μmol/L (interquartile range (IQR) 111, 396 μmol/L) versus median venous creatinine 212 μmol/L (IQR 114, 380 μmol/L). The median bias for creatinine was +1.0 (IQR -3.0, +8.0). Limits of agreement on Bland-Altman plot were +36.5μmol/L and – 34.4μmol/L (Figure 1). Overall, there remained a strong positive correlation between POC-Cr and IDMS-traceable enzymatic creatinine assays (R = 0.997; P&amp;lt;0.0001) (Figure 2). Median POC-K was 4.3 mmol/L, IQR 4.0, 4.8 mmol/L versus median venous potassium 5.0, IQR 4.3, 5.3 mmol/L. The median bias for potassium was +0.4 (IQR +0.2, +0.5). Limits of agreement on Bland-Altman plot were +0.16mmol/L and – 0.91mmol/L (Figure 3). A strong positive correlation between POC-K and laboratory potassium assays was also confimed (R = 0.937; P&amp;lt;0.0001) (Figure 4). Conclusion There was strong positive correlation between POC and laboratory analysis of venous creatinine, comparable to product literature (R = 0.997 vs 0.998). Limits of agreement were outside the CLIA criteria for acceptable performance (+/- 27μmol/L). POC and laboratory potassium levels were also strongly correlated and comparable to product literature (R = 0.937 vs 0.997). Limits of agreement were outside the CLIA criteria for acceptable performance (+/- 0.5mmol/L). This discrepancy is likely due to haemolysis in some blood samples and small sample size. Overall, this pilot study demonstrated a strong correlation between POC and laboratory testing. Larger scale evaluation is still required in a multi-ethnic population, prior to assessing feasibility in community-led protocolized pathways.