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Hemostat Thromboplastinliquid Manual Testing on HumaClot Junior and HumaClot DuoPlus

Perform duplicate testing of samples and controls.
Determination of Prothrombin Time (PT) HumaClot Junior
Package Size HumaClot DuoPlus
[REF] 31012 6 x 2 ml Pre-warm [RGT] to 37°C before use and pre-warm test tubes.
[IVD] Pipette Plasma/ Control into pre-warmed test 100 μl
tube.
Intended Use
Incubate 3 min. at 37°C
HEMOSTAT THROMBOPLASTINliquid is a highly sensitive reagent intended
for the manual or automated determination of prothrombin time (PT). Add pre-warmed [RGT] 200 μl
Prolongation of the PT indicates either acquired or congenital disorders Start timer with addition of [RGT]. Record time required for clot
that affect coagulation factors I, II, V, VII and X. The PT has been widely formation.
accepted as the means to monitor patients on anticoagulant therapy, due
to the reduction in the activity of vitamin K dependent clotting factors (II, Quality Control
VII, IX, X, Protein C and S). HEMOSTAT THROMBOPLASTINliquid can be used Routine quality control is indispensable in coagulation testing.
to assay coagulation factors in the extrinsic and common pathways of HEMOSTAT CONTROL PLASMA NORMAL (CPN) and HEMOSTAT CONTROL
coagulation. PLASMA ABNORMAL (CPA) should be run in conjunction with patient
Test Principle plasmas. CPN is a normal plasma, whereas CPA is adjusted to mimic
moderately deficient plasmas.
The one-stage PT measures the clotting time of plasma after adding a
source of tissue factor (thromboplastin) and calcium. The recalcification of Results
plasma in the presence of tissue factor generates activated factor Xa, with The results can be reported in seconds, in Prothrombin ratio, in % of norm
the consequent formation of thrombin and ultimately an insoluble fibrin or as INR.
clot.
Seconds: Calculate the mean time of duplicate PT determination for each
Content plasma and report to the nearest 0.1 second.
[RGT] 6 x 2 ml Thromboplastin reagent (liquid) Prothrombin Ratio (PR): To obtain the Prothrombin ratio, the reaction
Rabbit brain extract >10% time of the sample is divided by the reaction time of the normal plasma
CaCl2 0.2% pool.
Sodium azide <0.01%
PTpatient [s]
Material required but not provided PR = 
[REF] 35500 HEMOSTAT CALIBRATOR PTnormal plasma pool [s]
[REF] 35001 HEMOSTAT CONTROL PLASMA NORMAL HEMOSTAT CALIBRATOR (please refer to the table “Examples of
Reconstitution” in the HEMOSTAT CALIBRATOR instruction for use (IFU))
[REF] 35002 HEMOSTAT CONTROL PLASMA ABNORMAL
can be used as a normal plasma pool. Alternatively, a normal plasma pool
Sodium chloride 0.9 % (0.154 mol/l) of apparently “healthy” donors can be prepared.
Reagent Preparation % of norm: To report results in % of norm a calibration curve is required.
No special preparation step is necessary as this is a liquid reagent, ready to Prepare a calibration curve using HEMOSTAT CALIBRATOR. Alternatively
use. Let the thromboplastin stand at 18...25°C for 30 minutes. Swirl the the calibration curve can be generated with a normal plasma pool
vial gently, horizontally several times (5-10) before use, but do not shake. prepared from apparently “healthy” donors, declaring the Prothrombin
time as 100%.
Storage and Stability
When stored at 2...8°C unopened vials are stable until the expiry date. Establishment of the Reference Curve - Fully Automated Testing:

After opening of [RGT]
Temperature 37°C 20…25°C 15…19°C 2…8°C
Time 2 days 4 days 5 days 12 days
Store at 2...8°C when not in use. Do not freeze!
Specimen Collection
Use only plastic or siliconised glass containers and use 3.2% (0.109 M)
buffered sodium citrate as anticoagulant (sodium oxalate, EDTA and
heparin are not suitable). The use of higher concentration of trisodium
citrate (3.8 %; 129 mmol/l) is not recommended.
Obtain venous blood by clean venipuncture. Immediately mix 9 parts
blood with 1 part anticoagulant. Avoid foaming the specimen.
Specimen Processing
Centrifuge for 15 minutes at 1500 x g.
Remove plasma using a plastic pipette and store in a plastic tube. Cover
specimens to prevent pH changes that may affect test results. Turbid,
icteric, lipemic, or haemolysed specimens may generate erroneous results.
Specimens maintained at 18...25°C must be tested within 24 hours. Do
not store at 2...8°C, because it may undergo cold activation resulting in
significant shortening of the PT. For longer periods of time, plasma should
be frozen at -20°C for up to 2 weeks or at -70°C for up to 12 months. Thaw
samples rapidly at 37°C, swirl gently and test immediately. Do not re-
freeze.
Test Procedure
HEMOSTAT THROMBOPLASTINliquid can be used manually or on automated
coagulation analyzers.
Fully Automated Testing and Manual Testing on HumaClot Junior model
HC1, HumaClot Duo model HC2 and HumaClot Quattro
The Application Sheets can be accessed via www.human.de/aps-coag/
If the Application Sheets are not accessible via internet, they can be
obtained free of charge from your local distributor. The Application Sheets
contain analyzer/assay specific handling and performance information.
Please also consult the user manual of the respective instrument.
Refer to the HEMOSTAT CALIBRATOR IFU and the user manual or the
Application Sheets of the instrument.
Establishment of the Reference Curve – Manual Testing:
Generate a calibration curve diluting the HEMOSTAT CALIBRATOR (for
correct reconstitution please refer to the table “Examples for
Reconstitution” in the HEMOSTAT CALIBRATOR IFU) or a normal plasma
pool with sodium chloride (0,154 mol/l):
Dilution Scheme
HEMOSTAT CALIBRATOR undiluted 1+1 1+3 1+7
or normal plasma pool
% of norm 100% 50% 25% 12.5%
INR: The interdependency of the Prothrombin time of the used
Thromboplastin and instrument can be corrected through the
determination of the “International Normalized Ratio (INR). For this
purpose the Prothrombin ratio can be converted into internationally
comparable values by means of the “International Sensitivity Index (ISI)”.
INR = PRISI
The ISI-value for HEMOSTAT THROMBOPLASTINliquid is stated in the lot-
specific Table of Assigned Values.
Reference Range
For normal plasma a range of PT = 10-14 seconds can be expected.
Each laboratory should establish its own normal range using instrumenta-
tion, blood collection methods, and testing techniques commonly used in
that laboratory. The normal range should be re-established with any
change in instrumentation, blood collection techniques, anticoagulant
and should be at least verified when changing to new lots of reagents.
Limitations
The PT may be prolonged by substances including corticosteroids, EDTA,
oral contraceptives, asparaginase, clofibrate, erythromycin, ethanol,
tetracycline and anticoagulants, such as heparin and warfarin.
The PT may be shortened by substances including antihistamines, bu-
tabarbital, caffeine, oral contraceptives, phenobarbital and vitamin K.
Performance Characteristics
Typical performance data can be found in the Verification Report, at
www.human.de/data/gb/vr/co-ptli.pdf or
www.human-de.com/data/gb/vr/co-ptli.pdf
If the performance data are not accessible via internet, they can be
obtained free of charge from your local distributor.
Safety Notes
[RGT]
P260 Do not breathe dust/fume/gas/mist/vapours/spray.
P262 Do not get in eyes, on skin, or on clothing.
P281 Use personal protective equipment as required.
P303+P361+P353 IF ON SKIN (or hair): Take off immediately all
contaminated clothing. Rinse skin with water/shower.
P305+P351+P338 IF IN EYES: Rinse cautiously with water for several
minutes. Remove contact lenses, if present and easy to do. Continue
rinsing.
P337+P313 If eye irritation persists: Get medical advice/attention.
P401 Store in accordance with local/regional/national/international
regulations.
P501 Dispose of contents/container in accordance with local/regional/
national/ international regulations.

CO-PTLI INF 3101201 GB 01-2020-007 |

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