Which is the best anticoagulant for whole blood aggregometry platelet function testing? Comparison of six anticoagulants and diverse storage conditions

Abstract

Major hindrances of impedance aggregometry are caused by limited storage time and the requirement of ex vivo anticoagulation. Data on the influence of different anticoagulants and storage conditions are rare and incomplete. This study has systematically examined the influence of six different anticoagulants (sodium and lithium heparin, 20 µg/mL and 45 µg/mL r-hirudin, benzylsulfonyl-D-Arg-Pro-4-amidinobenzylamide (BAPA), and citrate) on the results of Adenosine 5'-diphosphate (ADP) and arachidonic acid (AA) induced measurements using multiple-electrode impedance aggregometer (MEA). Measurements were carried out in a time frame of 0 min up to 48 h after blood withdrawal. In addition, the influence of storage temperatures of 4°C and 37°C was evaluated. Results of ADP-induced tests significantly varied within the first 30 min in all tested anticoagulants, in citrated blood even within the first 60 min. They remained stable up to 2 h in 20 µg/mL r-hirudin and BAPA, 4 h in citrate, 8 h in 45 µg/mL r-hirudin, and lithium heparin and up to a maximum of 12 h in sodium heparin anticoagulated blood. The analysis of AA-induced tests revealed no significantly different results up to 6 h when BAPA was used, 8 h in lithium heparin, 20 µg/mL r-hirudin and citrate, 12 h in 45 µg/mL r-hirudin, and even 24 h in sodium heparin-anticoagulated samples. A storage temperature of either 4°C or 37°C in contrast to room temperature had a negative influence on the stability of results. In conclusion, sodium heparin and 45 µg/mL r-hirudin as anticoagulants guarantee the longest storage time for impedance aggregometry.

New-Generation Thromboelastography: Comprehensive Evaluation of Citrated and Heparinized Blood Sample Storage Effect on Clot-Forming Variables

“…The results of platelet-function measurements depend on many variables, such as the different assays and cutoff values and the diverse specific and unspecific stimulants used, as well as on preanalytic variables, such as the anticoagulants used, storage time, and temperatures. [35][36][37] Variable prevalence of low-antiplatelet response to acetylsalicylic acid and thienopyridines, ranging between 1% to 45%, have been reported. 36,38 One of the crucial first steps of sample preparation for all platelet function testing, including for the TEG platelet-mapping assay, is ex vivo anticoagulation.…”

“…[35][36][37] Variable prevalence of low-antiplatelet response to acetylsalicylic acid and thienopyridines, ranging between 1% to 45%, have been reported. 36,38 One of the crucial first steps of sample preparation for all platelet function testing, including for the TEG platelet-mapping assay, is ex vivo anticoagulation. Platelets are stabilized in vivo by such mediators as nitric oxide, prostaglandin I2, prostacyclin, and endothelial ADPase to prevent pathologic thrombus formation.…”

“…Platelets are stabilized in vivo by such mediators as nitric oxide, prostaglandin I2, prostacyclin, and endothelial ADPase to prevent pathologic thrombus formation. 36,39 Because these modulators are absent under in vitro conditions, platelets become easily activated in a timedependent manner, which influences hemostasis and augments artificially false-positive results from blood samples. A study of Multiplate (Roche Diagnostics, Mannheim, Germany) samples showed decreased aggregation over time, but those samples were collected in hirudin tubes.…”

Multiple electrode aggregometry in thrombocytopenic haemato-oncologic patients – Influence of haematologic variables

“…Reference values for Lithium Heparin are reported, as values for Sodium Heparin are not available from the manufacturer. Kaiser, et al showed that Sodium Heparin and Lithium Heparin anticoagulated samples give equal results for ADP in the first hour after sample collection [14]. During further storage Sodium Heparin conserved the results better than Lithium Heparin.…”

Stability of VerifyNow P2Y12 assay results with citrate anticoagulation as compared to hirudin anticoagulation over 20-min period

“…Other studies assessing the use of hirudin with Multiple Electrode Aggregometry (MEA) demonstrated stability in results obtained with hirudin up to 12 hours post sampling [7,8].…”