A Reference chart for clinical biochemical tests of hemolyzed serum samples

Ni, Jun; Zhu, Wenbo; Wang, Yanyang; Wei, Xuefei; Li, Jingjing; Lü, Peng; Zhang, Kui; Bai, Bing

doi:10.1002/jcla.23561

Cited by 11 publications

(14 citation statements)

References 23 publications

(45 reference statements)

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“…The US FDA considers stored RBC units with a haemolysis level less than 1% to be safe for transfusion. Exactly 1% haemolysis is roughly equivalent to a cell-free Hb concentration of 0.01 mM in the unit, which is lower than the final cell-free Hb concentration achieved in this study of $0.0157 mM observed post-wash [19]. One must, however, remember that the RBC units in this study were outdated (60-70 days old), which likely contributed to the higher final cell-free Hb concentration compared with literature values for washing non-expired RBC units [20].…”

Section: Hb Concentration In the Retentate And Permeate Decreases Ove...contrasting

confidence: 66%

Tangential flow filtration facilitated washing of human red blood cells: A proof‐of‐concept study

Allyn

Weigand

et al. 2022

Vox Sanguinis

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Background and Objectives: Red blood cell (RBC) units in hypothermic storage degrade over time, commonly known as the RBC storage lesion. These older RBC units can cause adverse clinical effects when transfused, as older RBCs in the unit lyse and release cell-free haemoglobin (Hb), a potent vasodilator that can elicit vasoconstriction, systemic hypertension and oxidative tissue injury after transfusion. In this study, we examined a novel method of washing ex vivo stored single RBC units to remove accumulated cellular waste, specifically cell-free Hb, using tangential flow filtration (TFF) driven by a centrifugal pump. Materials and Methods:The TFF RBC washing system was run under hypothermic conditions at 4 C, at a constant system volume with 0.9 wt% saline as the wash solution. The RBC washing process was conducted on 10 separate RBC units. For this proof-of-concept study, RBC units were expired at the time of washing (60-70 days old). Cell-free Hb was quantified by UV-visible absorbance spectroscopy and analysed via the Winterbourn equations. Pre-and post-wash RBC samples were analysed by Hemox Analyser, Coulter counter and Brookfield rheometer. The RBC volume fraction in solution was measured throughout the wash process by standard haematocrit (HCT) analysis.Results: No substantial decrease in the HCT was observed during the TFF RBC washing process. However, there was a significant decrease in RBC concentration in the first half of the TFF RBC wash process, with no significant change in RBC concentration during the second half of the TFF cell wash process with an 87% overall cell recovery compared with the total number of cells before initiation of cell washing. Utilization of the extinction coefficients and characteristic peaks of each Hb species potentially present in solution was quantified by Winterbourn analysis on retentate and permeate samples for each diacycle to quantify Hb concentration during the washing process. Significant cell-free Hb reduction was observed within the first four diacycles with a starting cell-free Hb concentration in the RBC unit of 0.105 mM, which plateaus to a constant Hb concentration of 0.01 mM or a total extracellular Hb mass of 0.2 g in the resultant washed unit. The oxygen equilibrium curve showed a significant decrease in P 50 between the initial and final RBC sample Shuwei Lu and Megan Allyn contributed equally to the work.

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Section: Hb Concentration In the Retentate And Permeate Decreases Ove...contrasting

confidence: 66%

Tangential flow filtration facilitated washing of human red blood cells: A proof‐of‐concept study

Allyn

Weigand

et al. 2022

Vox Sanguinis

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“…However, the time-dependent rise in LDH activity was significant only at the three-hour time point when tested in a pairwise manner (Student t-test p-value 0 vs. 180 min (LDH activity) = 0.02, Figure 1B), rendering it a far less sensitive marker than GSH (Student t-test p-value 0 vs. 180 min 1.03 × 10 -9 , Figure 1B). Already, 0.3% of hemolyzed red blood cells result in a notable red coloring of bloodderived products due to hemoglobin release [21]. We did not observe any red coloring in plasma samples after 3 h RT incubation, but the increase in GSH corresponds to a release of ~4.3% of total erythrocyte GSH (at 0.42 erythrocyte volume fraction, 1.2 mmol/L GSH concentration).…”

Section: Discussionmentioning

confidence: 47%

Blood Plasma Quality Control by Plasma Glutathione Status

et al. 2021

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Timely centrifugation of blood for plasma preparation is a key step to ensure high plasma quality for analytics. Delays during preparation can significantly influence readouts of key clinical parameters. However, in a routine clinical environment, a strictly controlled timeline is often not feasible. The next best approach is to control for sample preparation delays by a marker that provides a readout of the time-dependent degradation of the sample. In this study, we explored the usefulness of glutathione status as potential marker of plasma preparation delay. As the concentration of glutathione in erythrocytes is at least two orders of magnitude higher than in plasma, even the slightest leakage of glutathione from the cells can be readily observed. Over the 3 h observation period employed in this study, we observed a linear increase of plasma concentrations of both reduced (GSH) and oxidized glutathione (GSSG). Artificial oxidation of GSH is prevented by rapid alkylation with N-ethylmaleimide directly in the blood sampling vessel as recently published. The observed relative leakage of GSH was significantly higher than that of GSSG. A direct comparison with plasma lactate dehydrogenase activity, a widely employed hemolysis marker, clearly demonstrated the superiority of our approach for quality control. Moreover, we show that the addition of the thiol alkylating reagent NEM directly to the blood tubes does not influence downstream analysis of other clinical parameters. In conclusion, we report that GSH gives an excellent readout of the duration of plasma preparation and the associated pre-analytical errors.

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“…Each of the plasma aliquots were spiked with lysed whole blood to form 0, 0.1, 0.2, 0.4, 1, 2, and 4% spikes, calculated via a hemolysis color chart ( n = 6 replicates) . The method of simulated hemolysis was adapted from previous studies. − …”

Section: Experimental Sectionmentioning

confidence: 99%

Comprehensive Lipidomic Workflow for Multicohort Population Phenotyping Using Stable Isotope Dilution Targeted Liquid Chromatography-Mass Spectrometry

et al. 2023

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Dysregulated lipid metabolism underpins many chronic diseases including cardiometabolic diseases. Mass spectrometry-based lipidomics is an important tool for understanding mechanisms of lipid dysfunction and is widely applied in epidemiology and clinical studies. With ever-increasing sample numbers, single batch acquisition is often unfeasible, requiring advanced methods that are accurate and robust to batch-to-batch and interday analytical variation. Herein, an optimized comprehensive targeted workflow for plasma and serum lipid quantification is presented, combining stable isotope internal standard dilution, automated sample preparation, and ultrahigh performance liquid chromatography-tandem mass spectrometry with rapid polarity switching to target 1163 lipid species spanning 20 subclasses. The resultant method is robust to common sources of analytical variation including blood collection tubes, hemolysis, freeze–thaw cycles, storage stability, analyte extraction technique, interinstrument variation, and batch-to-batch variation with 820 lipids reporting a relative standard deviation of <30% in 1048 replicate quality control plasma samples acquired across 16 independent batches (total injection count = 6142). However, sample hemolysis of ≥0.4% impacted lipid concentrations, specifically for phosphatidylethanolamines (PEs). Low interinstrument variability across two identical LC-MS systems indicated feasibility for intra/inter-lab parallelization of the assay. In summary, we have optimized a comprehensive lipidomic protocol to support rigorous analysis for large-scale, multibatch applications in precision medicine. The mass spectrometry lipidomics data have been deposited to massIVE: data set identifiers MSV000090952 and 10.25345/C5NP1WQ4S.

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A Reference chart for clinical biochemical tests of hemolyzed serum samples

Cited by 11 publications

References 23 publications

Tangential flow filtration facilitated washing of human red blood cells: A proof‐of‐concept study

Tangential flow filtration facilitated washing of human red blood cells: A proof‐of‐concept study

Blood Plasma Quality Control by Plasma Glutathione Status

Comprehensive Lipidomic Workflow for Multicohort Population Phenotyping Using Stable Isotope Dilution Targeted Liquid Chromatography-Mass Spectrometry

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