HORIBA hemostasis

Hemostasis Guide

Preanalytical Guidelines

Sample Tube



Potential Risk


Recommended Sodium citrate 3,2 % (109 mmol/l);
Acceptable Sodium Citrate 3,8% (0,129 M);
CTAD acceptable in some circumstances.


Serum or any other anticoagulant can lead to an incorrect result.





Sample dilution (excess CaCl2-Citratre binding)9




False results.

Sufficient volume.
Respect the required ratio of sodium citrate to whole blood (1:9).
Fill volume: ≥90 %.
Do not transfer from 1 tube to another.

Check expiry date of the tube.

Sample Collection


Potential Risk

Tube filling order:
Before filling citrate tube, discard the first tube (neutral or citrate).
Citrate tube: before tubes with additives.
Sample contamination;
Sample dilution (excess CaCl2-Citratre binding)⁹.


Potential Risk

Proper Tube Identification Label / patient demographic and collection date and time.
The diameter of the needle recommended should preferably be between 19 and 22 gauge¹¹.
Acceptable: 23 (pediatric, compromised veins, geriatrics, oncology)¹¹.

Wrong result;


Risk of contamination from tissue thromboplastin and hemolysis.

Fibrinolysis activation;
Acidosis (pH <7,3);
PT prolonged.

Coagulation activation⁴·⁷;
Interferences, sample dilution.


Factor activation, Hemolysis¹;
Sample clotting⁴·⁷ (partial);
Variable anticoagulant gradient (gradient of sample with different citrate buffering).

Vacuum tube holder: the venipuncture must be swift and the blood flow, regular¹¹.

Winged collection butterfly: before filling citrate tube, discard the first tube (neutral or citrate)¹¹.

Avoid traumatic phlebotomy (draw);
Avoid drip lines;
Avoid wet alcohol carryover.

Release the tourniquet immediately when the first tube starts to fill (<1 mn).




Immediately mix gently by 3 to 6 complete end-over-end inversions to ensure adequate mixing with anticoagulant and to prevent clottin.

Sample Transport


Potential Risk

Room temperature (15 – 25 °C): should be maintained.
Keep the tube vertical during transport.
Prohibits transport on ice or refrigerate transport (2°C - 8°C).
False results: activation of some coagulation factors.

Sample Stability


Potential Risk

Fresh sample: Room temperature (15 – 25 °C)¹³ = 4 hours for most tests.
NOTE: this specification is true for most routine tests, for details per parameter, refer to GFHT ².
False results: activation of some coagulation factors.

Centrifugation and Storage


Potential Risk

Standard recommendation: 1500 g, 15 minutes.
Centrifugal conditions must be established and validated by the laboratory.
Maximum time for centrifugation after sampling is 2 hours.
Room temperature (15 – 25 °C)¹³.
“Rapid centrifugation“ may be used (higher speed, shorter duration) under lab validation.

False results due to contamination by phospholipid from platelets;
Lupus and Heparin Assays particularly affected10;
PT, APTT, TT not affected up to Platelet count=200 000/μl ¹.

False results;
Lupus and Heparin Assays affected¹⁰.




FVII activation, platelet disruption, loss of coagulation components, Hemolysis¹.


False results due to the release of phospholipid.

Double centrifugation recommended before freezing.
Transfer the plasma to a non-activating plastic centrifuge tube using a plastic pipette, then re-centrifuging the sample for an additional 10 minutes. When transferring to a secondary tube, take care to not include any residual platelets that may have collected at the bottom of the centrifuge tube.

Storage (depending on the parameter)
Room temperature (15 – 25 °C)¹²,¹³ . 4 hours for most of tests.
Minus 20°C = Maximum storage time: 2 weeks.
Minus 70°C = 6 month to 12 month.

NOTE: these specifications are true for most of tests, for details per parameter, refer to GFHT².

Defreezing must be done at 37°C in bain-marie during 5 to 10 minutes maximum1.


Pre-analytical Sample Integrity


Potential Risk

Avoid clotted samples.
Check samples for Hemolysis, lipid and icterus.
Coagulation activation⁴ ⁶ ⁷ (micro clots could be not visible);
False results.


1 – Clinical Laboratory Standard Institute
CLSI. Collection t, and processing of blood specimens for testing plasma-based coagulation assays and molecular hemostasis assays; approved guideline. 5e edition. CLSI Document H21-A5. Ed. Wayne: PCaLSI, 2008.
CLSI. Procedures for the Handling and Processing of Blood Specimens for Common Laboratory Tests; Approved Guideline – Fourth Edition, CLSI document GP44-A4, Wayne, PA : CLSI, 2010, 57 p.
CLSI. Laboratory Testing for the Lupus Anticoagulant; Approved Guideline. CLSI Document H60-A. Wayne, PA: Clinical and Laboratory Standards Institute; 2014
CLSI. Quantitative D-dimer for the exclusion of venous thromboembolic disease; approved guideline. 1e edition. CLSI Document H59-A. Ed. Wayne: PCaLSI, 2016
CLSI. Collection of Diagnostic Venous Blood Specimens; Approved Standard- Seventh Edition, CLSI document GP41-A7, Wayne, PA: CLSI, 2017, 86 p.
2 – GFHT Recommandations pré-analytiques en hémostase : Stabilité des paramètres d’hémostase générale et délais de réalisation des examens - Mai 2017
3 – Töpfer et al. J Lab Med 2000; 24 : 514-20
4 – Lippi et al., Blood Coagul Fibrinolysis 2005; 16: 453-9
5 – Lippi et al., Clin Lab Haematol 2006; 28: 332-7
6 – Interference of Blood cell lysis on routine coagulation testing, G.Lippi et al, Arch pathol Lab Med, 2006
7 – Preanalytical Variables in Coagulation Testing Associated with diagnostic errors in Hemostasis, E.J. Favaloro, D. Funk, G.Lippi, Labmedicine, 2012
8 – Cattaneo M et al. J Thromb Haemost 2013; 11: 1183–9
9 – Preanalytical and Post analytical Variables: the leading causes of diagnostic error in Hemostasis?,E.J. Favaloro, G.Lippi, D.Adcock, Seminars in Thrombosis and Hemostasis Volume 34 Number 7, 2008 10 – Pengo V, Tripodi A, Reber G, et al. Update of the guidelines for lupus anticoagulant detection. Subcommittee on Lupus Anticoagulant/Antiphospholipid Antibody of the Scientific and Standardization Committee of the International Society on Thrombosis and Haemostasis. J Thromb Haemost. 2009; 7(10):1737-1740.
11 – Pre-analytical issues in the haemostasis laboratory: guidance for the clinical laboratories A. Magnette, M. Chatelain, B. Chatelain, H. Ten Cate, and F. Mullier Thromb J. 2016; 14: 49
12 – Toulon P et al. Impact of different storage times at room temperature of unspun citrated blood samples on routine coagulation tests results. Results of a bicenter study and review of the literature. Int J Lab Hematol 2017; 39:458–468. doi:10.1111/ijlh.12660
13 – Pharmacopée Européenne

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