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DOI: 10.1055/a-2749-6046
Laboratory Monitoring of UFH in Different Settings (DEXHEP Study): Association between Anti-Xa Levels, Platelet Factor 4 (PF4) Plasma Levels and Dextran Sulfate
Authors
Funding Information This work was supported by the Société Française de Thrombose et d'Hémostase (SFTH, French Society on Thrombosis and Haemostasis).

Abstract
Background
Chromogenic anti-Xa assay is currently used in the management of patients on unfractionated heparin (UFH). It has been shown that inter-assay variability in anti-Xa levels can be explained in part by the presence or absence of dextran sulfate (DXS) in the reagents. DXS has the ability to dissociate UFH from neutralizing proteins, including platelet factor 4 (PF4).
Aims
Investigate whether PF4 plasma levels along with the presence/absence of DXS in anti-Xa reagents are associated with variations in UFH anti-Xa levels in different clinical situations.
Methods
In the prospective multicenter study DEXHEP-NCT04700670, critically ill patients on UFH therapy (four groups) were recruited. Blood was collected into citrate and CTAD tubes. Chromogenic anti-Xa levels were assessed using seven reagent/analyzer combinations including two without DXS. Plasma PF4 was measured by ELISA (Zymutest-PF4-Hyphen-Biomed).
Results
A total of 144 patients were analyzed: average PF4 levels in citrate plasma samples were consistently higher than in CTAD ones (206 vs. 46 ng/mL, p < 10−4), regardless of the patient group. Using a linear mixed-effect model, we found a significant effect of both DXS and PF4 on anti-Xa level, with a significant interaction term (p < 10−4). Considering the 0.3 to 0.7 IU/mL therapeutic range, agreement between anti-Xa values (Liquid-anti-Xa/DXS-free vs. Biophen-LRT/DXS) was observed in roughly two-thirds of the patients.
Conclusion
PF4 levels slightly affects anti-Xa levels, the use of CTAD tubes minimizing the effect. However, PF4 levels do not fully explain the differences of anti-Xa levels observed in the presence or absence of DXS, which has a greater effect. Anti-Xa assays require better standardization.
Publication History
Received: 16 August 2025
Accepted after revision: 22 October 2025
Accepted Manuscript online:
18 November 2025
Article published online:
02 December 2025
© 2025. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)
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References
- 1 Garcia DA, Baglin TP, Weitz JI, Samama MM. Parenteral anticoagulants: antithrombotic therapy and prevention of thrombosis, 9th ed: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines. Chest 2012; 141 (2, Suppl): e24S-e43S
- 2 Smythe MA, Priziola J, Dobesh PP, Wirth D, Cuker A, Wittkowsky AK. Guidance for the practical management of the heparin anticoagulants in the treatment of venous thromboembolism. J Thromb Thrombolysis 2016; 41 (01) 165-186
- 3 Arachchillage DJ, Kitchen S. Pleiotropic effects of heparin and its monitoring in the clinical practice. Semin Thromb Hemost 2024; 50 (08) 1153-1162
- 4 Gouin-Thibault I, Mansour A, Hardy M. et al. Management of therapeutic-intensity unfractionated heparin: a narrative review on critical points. TH Open 2024; 8 (03) e297-e307
- 5 Gehrie E, Laposata M. Test of the month: the chromogenic antifactor Xa assay. Am J Hematol 2012; 87 (02) 194-196
- 6 Toulon P, Smahi M, De Pooter N. APTT therapeutic range for monitoring unfractionated heparin therapy. Significant impact of the anti-Xa reagent used for correlation. J Thromb Haemost 2021; 19 (08) 2002-2006
- 7 Lyon SG, Lasser EC, Stein R. Modification of an amidolytic heparin assay to express protein-bound heparin and to correct for the effect of antithrombin III concentration. Thromb Haemost 1987; 58 (03) 884-887
- 8 Mouton C, Calderon J, Janvier G, Vergnes MC. Dextran sulfate included in factor Xa assay reagent overestimates heparin activity in patients after heparin reversal by protamine. Thromb Res 2003; 111 (4-5): 273-279
- 9 Ignjatovic V, Summerhayes R, Gan A. et al. Monitoring unfractionated heparin (UFH) therapy: which anti-factor Xa assay is appropriate?. Thromb Res 2007; 120 (03) 347-351
- 10 Smahi M, De Pooter N, Hollestelle MJ, Toulon P. Monitoring unfractionated heparin therapy: lack of standardization of anti-Xa activity reagents. J Thromb Haemost 2020; 18 (10) 2613-2621
- 11 Hollestelle MJ, van der Meer FJM, Meijer P. Quality performance for indirect Xa inhibitor monitoring in patients using international external quality data. Clin Chem Lab Med 2020; 58 (11) 1921-1930
- 12 Amiral J, Amiral C, Dunois C. Optimization of heparin monitoring with anti-FXa assays and the impact of dextran sulfate for measuring all drug activity. Biomedicines 2021; 9 (06) 700
- 13 Toulon P, Smahi M, De Pooter N. APTT therapeutic range for monitoring unfractionated heparin therapy. Significant impact of the anti-Xa reagent used for correlation: response from original authors Toulon et al. J Thromb Haemost 2021; 19 (08) 2090-2091
- 14 Lasne D, Toussaint-Hacquard M, Delassasseigne C. et al. Factors influencing anti-Xa assays: a multicenter prospective study in critically ill and noncritically ill patients receiving unfractionated heparin. Thromb Haemost 2023; 123 (12) 1105-1115
- 15 Hardy M, Cabo J, Deliège A. et al. Reassessment of dextran sulfate in anti-Xa assay for unfractionated heparin laboratory monitoring. Res Pract Thromb Haemost 2023; 7 (08) 102257
- 16 Hammami E, Stiel L, Palpacuer C, Harzallah I. Heparin monitoring during extracorporeal membrane oxygenation: the effect of dextran sulfate on anti-Xa assay. Res Pract Thromb Haemost 2023; 7 (07) 102196
- 17 Warkentin TE. Classification of platelet-activating anti-platelet factor 4 disorders. Int J Lab Hematol 2025
- 18 Gritters-van den Oever M, Schoorl M, Schoorl M, Bartels PC, Grooteman MP, Nubé MJ. The role of the extracorporeal circuit in the trapping and degranulation of platelets. Blood Purif 2009; 28 (03) 253-259
- 19 Mazzeffi M, Clark M, Grazioli A. et al. Platelet factor-4 concentration in adult veno-arterial ECMO patients. Perfusion 2021; 36 (07) 688-693
- 20 Sancheti M, Rentschler M, Bolch C. et al. Designing an experimental method for assessing biocompatibility of circuit coatings using biomarkers for platelet activation during cardiopulmonary bypass. J Extra Corpor Technol 2024; 56 (02) 37-44
- 21 Contant G, Gouault-Heilmann M, Martinoli JL. Heparin inactivation during blood storage: its prevention by blood collection in citric acid, theophylline, adenosine, dipyridamole-C.T.A.D. mixture. Thromb Res 1983; 31 (02) 365-374
- 22 Kitchen S, Adcock DM, Dauer R. et al. International Council for Standardisation in Haematology (ICSH) recommendations for collection of blood samples for coagulation testing. Int J Lab Hematol 2021; 43 (04) 571-580
- 23 Kitchen S, Adcock DM, Dauer R. et al. International Council for Standardization in Haematology (ICSH) recommendations for processing of blood samples for coagulation testing. Int J Lab Hematol 2021; 43 (06) 1272-1283
- 24 R Core Team. (2021). R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria; . Accessed at: https://www.R-project.org/
- 25 Bates D, Maechler M, Bolker B, Walker S. Fitting linear mixed-effects models using lme4. J Stat Softw 2015; 67 (01) 1-48
- 26 Hindle MS, Cheah LT, Yates DM, Naseem KM. Preanalytical conditions for multiparameter platelet flow cytometry. Res Pract Thromb Haemost 2023; 7 (07) 102205
- 27 Toulon P, Abecassis L, Smahi M, Ternisien C. Monitoring treatments with unfractionated heparin: CTAD must be used instead of citrate as the anticoagulant solution when using partial-draw collection tubes. Results of a multicenter evaluation. Thromb Res 2010; 126 (06) 536-542
- 28 Billoir P, Clavier T, Guilbert A. et al. Is citrate theophylline adenosine dipyridamole (CTAD) better than citrate to survey unfractionated heparin treatment? Has delayed centrifugation a real impact on this survey?. J Thromb Thrombolysis 2019; 48 (02) 277-283
- 29 Gremillet M, Talon L, Lebreton A, Sinegre T. Monitoring heparin therapy: stability of two different anti-Xa assays using blood samples collected in citrate-containing and CTAD tubes. Thromb J 2023; 21 (01) 21
- 30 Mayo KH, Ilyina E, Roongta V. et al. Heparin binding to platelet factor-4. An NMR and site-directed mutagenesis study: arginine residues are crucial for binding. Biochem J 1995; 312 (Pt 2): 357-365
- 31 Levine MN, Hirsh J, Gent M. et al. A randomized trial comparing activated thromboplastin time with heparin assay in patients with acute venous thromboembolism requiring large daily doses of heparin. Arch Intern Med 1994; 154 (01) 49-56