Stability of coagulation assays performed in plasma from citrated whole blood transported at ambient temperature

2008 ◽  
Vol 99 (02) ◽  
pp. 416-426 ◽  
Author(s):  
Manuel Zürcher ◽  
Irmela Sulzer ◽  
Gabriela Barizzi ◽  
Bernhard Lämmle ◽  
Lorenzo Alberio
Keyword(s):  
Ambient Temperature ◽  
Whole Blood ◽  
Repeated Measures ◽  
Blood Samples ◽  
Coagulation Assays ◽  
Specimen Collection ◽  
Collection Tube ◽  
Thrombophilia Screening ◽  
Clinically Significant ◽  
The Stability

SummaryMany preanalytical variables affect the results of coagulation assays. A possible way to control some of them would be to accept blood specimens shipped in the original collection tube. The aim of our study was to investigate the stability of coagulation assays in citrated whole blood transported at ambient temperature for up to two days after specimen collection. Blood samples from 59 patients who attended our haematology outpatient ward for thrombophilia screening were transported at ambient temperature (outdoor during the day, indoor overnight) for following periods of time: <1 hour, 4–6, 8–12, 24–28 and 48–52 hours prior to centrifugation and plasma-freezing. The following coagulation tests were performed: PT, aPTT, fibrinogen, FII:C, FV:C, FVII:C, FVIII:C, FIX:C, FX:C, FXI:C,VWF:RCo,VWF:Ag, AT, PC activity, total and free PS antigen, modified APC-sensitivity-ratio, thrombin-antithrombin-complex and D-dimer. Clinically significant changes, defined as a percentage change of more than 10% from the initial value, were observed for FV:C, FVIII:C and total PS antigen starting at 24–28 hours, and for PT, aPTT and FVII:C at 48–52 hours. No statistically significant differences were seen for fibrinogen, antithrombin, or thrombin-antithrombin complexes (Friedman repeated measures analysis of variance).The present data suggest that the use of whole blood samples transported at ambient temperature may be an acceptable means of delivering specimens for coagulation analysis. With the exception of factorV andVIII coagulant activity, and total PS antigen all investigated parameters can be measured 24–28 hours after specimen collection without observing clinically relevant changes.

scholarly journals Stability Studies of Twenty-Four Analytes in Human Plasma and Serum

2002 ◽  
Vol 48 (12) ◽  
pp. 2242-2247 ◽  
Author(s):  
Bobby L Boyanton ◽  
Kenneth E Blick
Keyword(s):  
Blood Cells ◽  
Repeated Measures ◽  
Room Temperature ◽  
Stability Studies ◽  
Pump Failure ◽  
Glucose Depletion ◽  
Prolonged Contact ◽  
Biochemical Mechanisms ◽  
Clinically Significant ◽  
The Stability

Abstract Background: The stability and stoichiometric changes of analytes in plasma and serum after prolonged contact with blood cells in uncentrifuged Vacutainer® tubes were studied. Methods: We simultaneously investigated the stability of 24 analytes (a) after prolonged contact of plasma and serum with blood cells and (b) after immediate separation of plasma and serum (centrifuged twice at 2000g for 5 min). We verified biochemical mechanisms of observed analyte change by concomitant measurement of pH, Pco2, and Po2. Hemolysis was qualitatively and semiquantitatively assessed. All specimens were maintained at room temperature (25 °C) and analyzed in duplicate 0.5, 4, 8, 16, 24, 32, 40, 48, and 56 h after collection. Statistically significant changes from the 0.5 h mean were determined using repeated-measures ANOVA. The significant change limit was applied to determine clinically significant changes in measured analytes. Results: Fifteen of 24 analytes in plasma and serum maintained in contact with cells showed clinically relevant changes, with the degree of change more pronounced in most plasma specimens. All analytes in plasma and serum immediately separated from cells after collection were stable. Conclusion: Storage of uncentrifuged specimens beyond 24 h caused significant changes in most analytes investigated because of (a) glucose depletion and Na+,K+-ATPase pump failure; (b) the movement of water into cells, causing hemoconcentration; and (c) leakage of intracellular constituents and metabolites. Immediate separation of plasma or serum from cells provides optimal analyte stability at room temperature. When prolonged contact of plasma or serum with cells is unavoidable, use of serum is recommended because of the higher instability of plasma analytes.

Short-term stability of free metanephrines in plasma and whole blood

2020 ◽  
Vol 58 (5) ◽  
pp. 753-757 ◽  
Author(s):  
Elisa Danese ◽  
Martina Montagnana ◽  
Claudio Brentegani ◽  
Giuseppe Lippi
Keyword(s):  
Whole Blood ◽  
Repeated Measures ◽  
Sample Collection ◽  
Short Term ◽  
Total Change ◽  
Term Stability ◽  
Before And After ◽  
Liquid Chromatography Tandem Mass ◽  
The Mean ◽  
The Stability

AbstractBackgroundAnalysis of plasma metanephrine (MN) and normetanephrine (NMN) with liquid chromatography tandem mass spectrometry (LC-MS/MS) is the gold standard for the screening of pheochromocytomas and paragangliomas (PPGLs). As scarce information is available on the stability of MNs in diagnostic samples, this study was aimed at analyzing the short-term stability of plasma free MNs in whole blood and plasma, using LC-MS/MS.MethodsThe stability of plasma MNs was evaluated after sample collection at 1, 2 and 3 h in whole blood, and at 2, 4 and 6 h in centrifuged samples. Both studies were performed while maintaining the samples at room temperature (RT) and at 4 °C. The ClinMass Complete Kit (Recipe, Munchen, Germany) was used for measuring MNs with LC-MS/MS (Nexera X2 UHPLC-4500MD Sciex). Differences from the baseline (T0) were assessed using repeated measures one-way ANOVA, Students’ paired t-test and a comparison of the mean percentage changes with the total change limit (TCL).ResultsStatistically significant differences from T0 were found for both MNs (p < 0.001) in whole blood stored at RT, and for NMN (p = 0.028) but not MN (p = 0.220) at 4 °C. The mean difference exceeded the TCL after 1 h and 3 h at RT for MN, and after 1 h at RT for NMN. Statistically significant differences from T0 were only observed in the plasma samples for NMN at RT (p = 0.012), but the variation was within the TCL.ConclusionsMN and NMN displayed different patterns of stability before and after centrifugation. Even short-time storage at RT in whole blood should hence be avoided.

Effect of transport conditions on the stability of biochemical markers in blood.

1989 ◽  
Vol 35 (12) ◽  
pp. 2313-2316 ◽  
Author(s):  
S E Hankinson ◽  
S J London ◽  
C G Chute ◽  
R L Barbieri ◽  
L Jones ◽  
...  
Keyword(s):  
Whole Blood ◽  
Apolipoprotein B ◽  
Biochemical Markers ◽  
Room Temperature ◽  
Density Lipoprotein ◽  
Alpha Tocopherol ◽  
Endogenous Hormones ◽  
Blood Samples ◽  
Whole Blood Samples ◽  
The Stability

Abstract We examined the stability of lipids, carotenoids, alpha-tocopherol, and endogenous hormones in plasma prepared from whole blood that had been mailed to a central location for processing. Initially, to simulate transport conditions, whole-blood samples were stored in the laboratory, either at room temperature or cooled, for up to 72 h before processing. In the latter samples, lipid concentrations changed up to 1.4% per day, carotenoids up to -5.5%, and hormones up to 9.5%. In a second study, analyte concentrations in plasma from cooled whole blood mailed via overnight courier were compared with those from plasma that had been immediately separated, frozen, and mailed via overnight courier. Concentrations of cholesterol, high-density lipoprotein subfraction 3, apolipoprotein B, and retinol were stable. Overall, for each marker except estradiol, the between-person variation was at least twice the within-person variation. In a third study, at least 340 micrograms of DNA was recovered from 30 mL of cool-shipped whole blood. Our results indicate that shipping whole-blood samples by overnight courier is feasible for assay of several biochemical markers of interest in epidemiological research.

scholarly journals Plasma Creatinine and the Effect of Delay in Separation of Samples

1992 ◽  
Vol 29 (3) ◽  
pp. 307-309 ◽  
Author(s):  
William G Simpson
Keyword(s):  
Whole Blood ◽  
Continuous Flow ◽  
Plasma Creatinine ◽  
Blood Samples ◽  
Whole Blood Samples ◽  
Clinically Significant ◽  
Dependent Error

There is a prevailing opinion that creatinine is an analyte which may be assayed with confidence in the plasma of ageing whole-blood samples. This is true of continuous flow methods employing dialysis, but in this study, where creatinine was measured in old samples by a kinetic Jaffé method on a discrete analyser, the concentration of creatinine was falsely raised. When plasma was left in contact with cells for 48 h, there was an apparent rise in the plasma creatinine by over 100 μmol/L compared with the 2 h concentration. This method-dependent error is therefore clinically significant.

Design and Development of a Unit-Use Cartridge for Coagulation Testing in Whole Blood

1999 ◽  
Author(s):  
Rhonda Cheadle ◽  
Andy Maczuszenko ◽  
Cindra Widrig Opalsky
Keyword(s):  
Blood Sample ◽  
Whole Blood ◽  
Clot Formation ◽  
Coagulation Cascade ◽  
Design And Development ◽  
Blood Samples ◽  
Coagulation Assays ◽  
Coagulation Testing ◽  
Functional Features ◽  
Whole Blood Samples

Abstract The following describes the development of a disposable cartridge for use at the patient bedside to perform traditional coagulation assays on fresh whole blood samples. The cartridge provides a means by which a blood sample can be metered and quantitatively mixed with reagents that activate the coagulation cascade. Clot formation is subsequently detected using a microfabricated sensor housed within the cartridge. The functional features of the cartridge and sensor are described.

High-Throughput Amplification and Detection of Human Erythrocyte Antigen (HEA) Nucleotide Polymorphisms From Leukocyte Reduced Red Blood Cell (RBC) Units: Implications for Minor RBC Antigen Inventory Management by Hospital Transfusion Services

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3348-3348
Author(s):  
Cassandra Josephson ◽  
John Roback ◽  
Robert Myers ◽  
Lisa Hallock ◽  
Irene DeMezzo ◽  
...  
Keyword(s):  
Blood Cell ◽  
Whole Blood ◽  
Human Erythrocyte ◽  
Dna Analysis ◽  
Blood Collection ◽  
Blood Samples ◽  
Allele Specific ◽  
Whole Blood Samples ◽  
Clinically Significant ◽  
Erythrocyte Antigen

Abstract Abstract 3348 Background: Technologies have recently been developed for rapid determination of extended human erythrocyte antigen (xHEA) phenotypes. For example, a semi-automated method using allele-specific oligonucleotides targeted against 32 clinically significant minor RBC antigens has been used to determine donor xHEA phenotypes from whole blood samples. This approach is currently used by blood collection centers and medical centers with blood collection facilities (both sites have access to linked donor whole blood samples). Broader access to xHEA information closer to the point-of-care (e.g. Transfusion Services at a Medical Center without a blood collection facility) may provide an opportunity to enhance patient care by more quickly and broadly providing units with xHEA phenotypes (Klapper et al., 2010.) However, transfusion services would need to use integrally attached segments for testing, and with leukoreduced (LR) RBC units these segments have very low numbers of white blood cells (WBC) (and therefore DNA), potentially limiting analysis. This study was performed to determine whether a HEA-elongation mediated multiplex assay in solution (HEA-eMAP-S) (Xin et al., 2010) could accurately genotype segments from LR-RBC units for 32 clinically significant minor RBC antigens. Methods: Segments from pre-storage LR-RBC units (American Red Cross), &lt; 14 days old, were obtained from a large tertiary care Children's Hospital in the Southeastern US and residual WBC were quantified by flow cytometry. DNA was extracted using an extraction method developed at BioArray SolutionS (BAS) using commercial reagents (Qiagen, Inc., Valencia, CA), and then amplified with the Universal Beadchip™ package (HEA LR-eMAP-S Beadchip™ Kits) which contains allele specific oligonucleotides directed to 32 clinically significant blood group antigens (c, C, e, E, V, VS, K, k, Kpa, Kpa, Jsa, Jsb, Jka, Jkb, Fya, Fyb, M, N, S, s, Lua, Lub, Dia, Dib, Coa, Cob, Doa, Dob, Joa, Hy, Yta, Ytb mutation for hemoglobin S). DNA analysis results were correlated with RBC storage solution, WBC filter type, and serologic minor RBC antigen phenotypes of the units. Results: 102 LR-RBC units from whole blood donations were studied, 74 /102 (73 %) stored in AS-1 and 28 /103 (27 %) in CPDA-1 solution. All AS-1 units were pre-storage LR with Fenwal Sepacell Flex Excel Filters and all CPDA-1 units were pre-storage LR with Whole Blood Fenwal Filters (Fenwal Inc. Lake Zurich, IL). All units demonstrated &lt; 5 × 106 WBC/unit with 47 % having &lt; 4 × 104 WBC/unit, which is at or below the limit of flow cytometric detection. Complete genotyping data was obtained from all samples. Ten samples showing initial indeterminate results on Diego and one for N antigens produced complete results after repeat testing. Fifty-four percent of units were serologically phenotyped for 1–8 antigens by the blood collection center; there was 100% correlation between predicted phenotype from DNA analysis and serology for these units. Conclusions: The HEA LR-eMAP-S DNA analysis can be applied to optimally pre-storage LR-RBC units yielding &gt; 99 % accuracy for all minor red blood cell antigens tested. The ability to perform this type of testing in a hospital transfusion service opens up new possibilities for transfusion services to select from their existing inventory and more efficiently allocate units to recipients with specific phenotypic requirements for RBC units. Disclosures: Josephson: Immucor: Speakers Bureau. DeMezzo:Immucor: Employment. Tanzi:Immucor: Employment. Enriquez:Immucor: Employment. Lin:Immucor: Employment. Hashmi:Immucor: Employment.

Long-Term Preservation of Whole Blood Samples for Flow Cytometry Analysis in Normal and HIV-Infected Individuals from Africa

1990 ◽  
Vol 1 (1) ◽  
pp. 38-45 ◽  
Author(s):  
Renu B Lal ◽  
Subhash K Hira ◽  
Rita R Dhawan ◽  
Peter L Perine
Keyword(s):  
Flow Cytometry ◽  
Whole Blood ◽  
Phenotypic Analysis ◽  
Blood Samples ◽  
Remote Areas ◽  
Immunodeficiency Virus ◽  
Whole Blood Samples ◽  
The Stability ◽  
Proliferation Assays ◽  
Lymphocyte Phenotypes

A whole blood method requiring less than 4 ml of heparinized blood was developed to assess the practicality of preparing whole blood samples that could be easily stored, transported and readily used to determine the lymphocyte phenotypes and proliferation responses of individuals from remote areas who are infected with the human immunodeficiency virus. Minor modifications in standard whole blood procedure for lymphocyte phenotyping have significantly increased the stability of light scatter and fluorescence intensity of the cells for subsequent flow cytometry (FC) analysis. These changes include removal of lysis solution prior to fixation, fixation of monoclonal antibody-stained cells in 1% paraformaldehyde for 30 minutes and storage of fixed samples in medium containing 1% bovine serum albumin. Lymphocyte subsets and their functional subsets could reliably be determined on samples stored for up to 4 weeks. Further, blood samples could be kept at room temperature for up to 96 hours or at ambient temperature during transportation from Africa before staining for FC without affecting their quantitation. While samples could be processed for FC analysis under field-laboratory conditions, proliferation assays could only be performed on samples that were transported within 48 hours of their collection. The whole blood method saves time and expense and decreases the volumes of blood required to perform phenotypic analysis and functional assays on specimens collected in remote areas.

scholarly journals Assessment of a portable lactate meter for field use in the white rhinoceros (Ceratotherium simum)

2017 ◽  
Vol 84 (1) ◽  
Author(s):  
Georgina C. Cole ◽  
Adrian S.W. Tordiffe ◽  
Gerhard Steenkamp
Keyword(s):  
Blood Lactate ◽  
Whole Blood ◽  
Laboratory Method ◽  
Plasma Lactate ◽  
Potassium Oxalate ◽  
Limits Of Agreement ◽  
Blood Samples ◽  
White Rhinoceros ◽  
The Stability ◽  
Over Time

Blood lactate is a predictor of mortality in critically ill humans and animals. Handheld lactate meters have the potential to be used in the field to evaluate the condition of severely injured rhinoceroses but have not been compared with laboratory-based methods. Agreement between a handheld lactate meter and a laboratory method was assessed, as was the stability of rhino blood lactate in the anticoagulant sodium fluoride/potassium oxalate (fluoride/oxalate). Blood samples were obtained from 53 white rhinos that had been immobilised for management reasons. Lactate was measured by means of a handheld meter using whole blood in heparin (WBHEP), whole blood in fluoride/oxalate (WBFO) and fluoride/oxalate plasma (PFO). Results were recorded in both blood (BL) and plasma (PL) modes and compared to an established laboratory method for measuring plasma lactate. To assess the stability of lactate over time, blood lactate in fluoride/oxalate was measured on the handheld meter at intervals for up to 91 h. Agreement was best using WBFO in PL mode, with small bias (-0.16), tight 95% limits of agreement (LOA) (-1.46, 1.14) and a Pc (95% CI) of 0.97 (0.92, 0.99). The agreement was improved for all sample types when using the PL mode compared to the blood lactate (BL) mode. Blood lactate was stable in fluoride/oxalate for 91 h, with a mean change from baseline of 0.15 (-0.178, 0.478) mmol/L (mean, 95% CI). The handheld meter was found to be suitable for field use in white rhinos but provided more reliable results with the device in PL mode. Furthermore, rhino blood lactate was found to be stable in fluoride/oxalate for as long as 3 days.

Sign in / Sign up

Export Citation Format

Share Document