NIH, National Cancer Institute, Division of Cancer Treatment and Diagnosis (DCTD) NIH - National Institutes of Health National Cancer Institute DCTD - Division of Cancer Treatment and Diagnosis

Monitoring heparin therapy by the activated partial thromboplastin time--the effect of pre-analytical conditions.

Author(s): van den Besselaar AM, Meeuwisse-Braun J, Jansen-Grüter R, Bertina RM

Publication: Thromb Haemost, 1987, Vol. 57, Page 226-31

PubMed ID: 3603414 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to observe the effects of several pre-analytical variables on activated partial thromboplastin time (APTT) of blood specimens obtained from normal donors and patients receiving heparin treatment.

Conclusion of Paper

APTT decreased with Cephotest reagent, or low speed or short centrifugation. Storage of specimens at room temperature prior to or after centrifugation when sodium citrate or HEPES-buffered citrate were used as anticoagulants also decreased APTT. Storage of blood specimens at 4 degrees C generally led to increased APTT. Storage of CTAD specimens at room temperature had the least overall impact on APTT and therefore is preferable to other storage conditions for monitoring of heparin therapy.

Studies

  1. Study Purpose

    The purpose of this study was to determine the effects of different reaction solutions used when measuring APTT as well as the effects of anticoagulant on blood specimens from normal donors. Specimens were stored at room temperature and centrifuged at 940 x g for 30 minutes. APTT was determined no longer than 4 hours after venepuncture.

    Summary of Findings:

    HEPES-buffered citrate plasma had a slightly higher APTT with Manchester reagent than citrate plasma did (p<0.05), but no other significant differences were found between anticoagulants. The choice of reaction solution significantly altered APTT with specimens measured using the Manchester reagent having the longest average APTTs and the widest spread while those measured with Cephotest had the shortest APTTs and the smallest spread (p<0.05).

    Biospecimens
    Preservative Types
    • None (Fresh)
    Diagnoses:
    • Normal
    Platform:
    AnalyteTechnology Platform
    Morphology Hematology/ auto analyzer
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Acquisition Anticoagulant HEPES-buffered citrate
    Sodium citrate
    Citrate-theophylline-adenosine-dipyridamole
    Hematology/ auto analyzer Specific Reaction solution Actin-FS
    Automated APTT
    Cephotest
    Manchester reagent
  2. Study Purpose

    The purpose of this study was to determine the effects of centrifugation speed (600 x g, 940 x g, or 2200 x g), and duration (10 min instead of 30 min) on APTTs in specimens from patients receiving heparin. Measurement of APTTs took place 85, 200, or 315 minutes after venepuncture. Centrifugation and storage were at 4 degrees C.

    Summary of Findings:

    Low speed centrifugation (600 x g) and reduction of centrifugation time from 30 to 10 minutes each resulted in shorter APTTs (not always significantly) when compared to centrifugation at 940 x g for 30 minutes. Faster centrifugation then 940 x g did not significantly alter APTT.

    Biospecimens
    Preservative Types
    • None (Fresh)
    Diagnoses:
    • Other diagnoses
    Platform:
    AnalyteTechnology Platform
    Morphology Hematology/ auto analyzer
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Storage duration 85 min
    200 min
    315 min
    Storage Storage temperature 4 degrees C
    Biospecimen Aliquots and Components Centrifugation Multiple durations compared
    Multiple speeds compared
  3. Study Purpose

    The purpose of this study was to determine the effects of anticoagulant, collection tube type, and storage prior to or after centrifugation on APTT of specimens from patients receiving heparin. Centrifugation and storage took place at room temperature.

    Summary of Findings:

    APTT decreased steadily when centrifuged or uncentrifuged blood was stored at room temperature and sodium citrate was used as the anticoagulant. However, the decrease in APTT was smaller when storage occurred after centrifugation. HEPES-buffered citrate specimens showed decreases in APTT with storage that were comparable to sodium citrate, however, starting APTTs were about 10% shorter. Blood collected in CTAD and stored prior to centrifugation showed less shortening of APTT than when collected in sodium citrate and APTT stayed relatively unchanged when storage occurred after centrifugation in these tubes. APTT increased over time when plasma was stored in polystyrene tubes at room temperature. In contrast, APTT decreased with storage when plasma was stored in siliconized vacutainer tubes.

    Biospecimens
    Preservative Types
    • None (Fresh)
    Diagnoses:
    • Other diagnoses
    Platform:
    AnalyteTechnology Platform
    Morphology Hematology/ auto analyzer
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Storage conditions Plasma
    Uncentrifuged blood
    Centrifuged blood
    Storage Time at room temperature 30 min
    1 h
    3 h
    5 h
    Biospecimen Acquisition Anticoagulant Citrate-theophylline-adenosine-dipyridamole
    HEPES-buffered citrate
    Sodium citrate
    Biospecimen Acquisition Type of collection container/solution Polystyrene
    Vacutainer
    Biospecimen Aliquots and Components Centrifugation Centrifugation delays investigated
  4. Study Purpose

    The purpose of this study was to determine the effects of storage temperature and duration on APTT of specimens from patients receiving heparin.

    Summary of Findings:

    Storage of blood specimens at 4 degrees C with either sodium citrate or CTAD increased APTT (p<0.05). At room temperature, sodium citrate specimens showed a significant decrease in APTT (p<0.05) with a decline in APTT apparent by 3 h. In contrast, APTT remained relatively constant for CTAD specimens over the storage period.

    Biospecimens
    Preservative Types
    • None (Fresh)
    Diagnoses:
    • Other diagnoses
    Platform:
    AnalyteTechnology Platform
    Morphology Hematology/ auto analyzer
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Storage temperature 22 degrees C
    4 degrees C
    Storage Storage duration 1-1.5 h
    3-3.5 h
    5-5.5 h
    Biospecimen Acquisition Anticoagulant Citrate-theophylline-adenosine-dipyridamole
    Sodium citrate
  5. Study Purpose

    The purpose of this study was to determine the effects of storage and anticoagulant on plasma platelet factor 4 (PF4) concentrations in two normal blood specimens.

    Summary of Findings:

    PF4 concentrations increased with storage and were higher with storage at room temperature compared to 4 degrees C. PF4 concentrations were significantly higher in sodium citrate specimens than in either CTAD or Thrombotect specimens.

    Biospecimens
    Preservative Types
    • None (Fresh)
    Diagnoses:
    • Normal
    Platform:
    AnalyteTechnology Platform
    Protein Hematology/ auto analyzer
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Storage temperature Room temperature
    4 degrees C
    Storage Storage duration 1 h
    5 h
    Biospecimen Acquisition Anticoagulant Citrate-theophylline-adenosine-dipyridamole
    Sodium citrate
    Thrombotect

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