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Stability of plasma adrenocorticotrophic hormone (ACTH): influence of hemolysis, rapid chilling, time, and the addition of a maleimide.

Author(s): Livesey JH, Dolamore B

Publication: Clin Biochem, 2010, Vol. 43, Page 1478-80

PubMed ID: 20875812 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of rapid chilling and preservation with N-phenyl maleimide (NPM) on measurement of plasma adrenocorticotrophic hormone (ACTH) in specimens with varying amounts of hemolysis.

Conclusion of Paper

ACTH and insulin levels were decreased in a dose dependent manner by hemolysis (p<0.0001) regardless of processing method. However, specimens that were preserved with NPM and chilled on ice for 30 minutes prior to centrifugation and freezing (method D) showed the smallest loss of ACTH and insulin. 90% of ACTH was retained in the 1.0% hemolysis specimen despite storage at room temperature for 1 h prior to analysis. Specimens chilled in ice water for 30 minutes prior to centrifugation and freezing and analyzed immediately after thaw (method B) resulted in smaller decreases in ACTH and insulin than specimens which were immediately centrifuged and analyzed right after thaw (method A). Specimens that were left at room temperature for 1 h after thaw and not preserved with NPM (method C) had the greatest loss of ACTH and insulin. Insulin recoveries were greater than ACTH recoveries for all processing methods except with the addition of NPM. Further, the calculated amount of hemolysis that resulted in a 10% change in ACTH or insulin measurement was dependent on processing method and analyte.

Studies

  1. Study Purpose

    The purpose of this study was to determine the effects of rapid chilling, storage at room temperature, and preservation with NPM on measurement of plasma ACTH in specimens with varying amounts of hemolysis. Differing degrees of hemolysis were attained by adding whole blood that had been frozen at -28 degrees C for 30 minutes.

    Summary of Findings:

    Hemolysis decreased ACTH and insulin levels in a dose dependent manner (p<0.0001) regardless of processing. However, method D showed the smallest loss of ACTH and insulin. 90% of ACTH was retained in the 1.0% hemolysis specimen despite storage at room temperature for 1 h prior to analysis. Processing method B resulted in smaller decreases in ACTH and insulin than method A. Method C had the greatest loss of ACTH and insulin. Insulin recoveries were greater than ACTH recoveries for all processing methods except method D. Further the calculated amount of hemolysis that resulted in a 10% change in ACTH or insulin measurement was dependent on processing method and analyte.

    Biospecimens
    Preservative Types
    • Frozen
    • Other Preservative
    Diagnoses:
    • Normal
    Platform:
    AnalyteTechnology Platform
    Peptide Clinical chemistry/auto analyzer
    Protein Clinical chemistry/auto analyzer
    Protein Hematology/ auto analyzer
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Post-thaw duration 0 min
    1 h
    Storage Storage temperature In ice water
    Storage Storage duration 0 min
    30 min
    Biospecimen Aliquots and Components Biospecimen components 0% hemolyzed blood
    0.1% hemolyzed blood
    0.25% hemolyzed blood
    0.5% hemolyzed blood
    1.0% hemolyzed blood
    Biospecimen Preservation Type of fixation/preservation Frozen
    N-phenyl maleimide
    Biospecimen Aliquots and Components Hemolysis Freeze/thaw-induced
    Hemolysate added
    No hemolysate added
    View more

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