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

The effect of storage time and freeze-thaw cycles on the stability of serum samples.

Author(s): Cuhadar S, Koseoglu M, Atay A, Dirican A

Publication: Biochem Med (Zagreb), 2013, Vol. 23, Page 70-7

PubMed ID: 23457767 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to determine the effects of up to 10 freeze-thaw cycles and up to 3 months of storage at -20 degrees C on the levels of 17 clinical chemistry analytes in serum from out-patients.

Conclusion of Paper

Compared to fresh specimens, a single freeze-thaw cycle led to decreased total bilirubin and total protein, and 2 freeze-thaw cycles led to increased uric acid (UA) levels. Blood urea nitrogen (BUN) and calcium were stable for up to 3 freeze-thaw cycles, lactate dehydrogenase (LD) was stable for up to 5 freeze-thaw cycles, and albumin was stable for up to 7 freeze-thaw cycles. Compared to fresh serum, serum stored for 3 months at -20 degrees C had statistically and clinically significant elevations in total protein and BUN levels and decreases in albumin levels.

Studies

  1. Study Purpose

    The purpose of this study was to determine the effects of up to 10 freeze-thaw cycles on the levels of 17 clinical chemistry analytes in serum from out-patients. Specimens were thawed at room temperature for 1 h before assay or refreezing and thawing the following day. Due to multiple testing, statistical significance was defined as p<0.005 compared to fresh specimens.

    Summary of Findings:

    After a single freeze-thaw cycle, total bilirubin and total protein levels were lower than in fresh specimens (p<0.001 and p=0.004, respectively). UA was higher after 2 freeze-thaw cycles than in fresh specimens (p=0.001). Further, BUN and calcium were stable for up to 3 freeze-thaw cycles, LD was stable for up to 5 freeze-thaw cycles, and albumin was stable for up to 7 freeze-thaw cycles. While levels of triglycerides increased and creatinine kinase (CK) decreased after 6 and 8 freeze-thaw cycles, respectively, the changes were clinically irrelevant. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyl transferase (GGT), cholesterol, glucose, creatinine, direct bilirubin, and high density lipoprotein (HDL) cholesterol levels were unaffected by up to 10 free-thaw cycles.

    Biospecimens
    Preservative Types
    • Frozen
    • None (Fresh)
    Diagnoses:
    • Not specified
    Platform:
    AnalyteTechnology Platform
    Carbohydrate Clinical chemistry/auto analyzer
    Electrolyte/Metal Clinical chemistry/auto analyzer
    Lipid Clinical chemistry/auto analyzer
    Protein Clinical chemistry/auto analyzer
    Small molecule Clinical chemistry/auto analyzer
    Steroid Clinical chemistry/auto analyzer
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Storage Freeze/thaw cycling 0 cycles
    1 cycle
    2 cycles
    3 cycles
    4 cycles
    5 cycles
    6 cycles
    7 cycles
    8 cycles
    9 cycles
    10 cycles
    Biospecimen Preservation Type of fixation/preservation Frozen
    None (fresh)
  2. Study Purpose

    The purpose of this study was to determine the effects of up to 3 months of storage at -20 degrees C on the levels of 17 clinical chemistry analytes in serum from out-patients. Due to multiple testing, statistical significance was defined as p<0.013 compared to fresh specimens.

    Summary of Findings:

    Compared to fresh serum, serum stored for 3 months at -20 degrees C had statistically and clinically significant elevations in total protein and BUN levels. Albumin levels were non-significantly higher after 2 months at -20 degrees C but significantly lower after 3 months at -20 degrees C (p=0.001). While statistically significant decreases in ALT, AST, CK, GGT, and LD, and increases in UA were observed after storage at -20 degrees C compared to fresh specimens, the variations did not exceed the maximum desirable bias and were considered clinically irrelevant. Changes in creatinine, calcium, and BUN levels after storage at -20 degrees C compared to fresh specimens exceeded the maximum desirable variability but did not reach statistical significance.

    Biospecimens
    Preservative Types
    • Frozen
    • None (Fresh)
    Diagnoses:
    • Not specified
    Platform:
    AnalyteTechnology Platform
    Carbohydrate Clinical chemistry/auto analyzer
    Electrolyte/Metal Clinical chemistry/auto analyzer
    Lipid Clinical chemistry/auto analyzer
    Protein Clinical chemistry/auto analyzer
    Small molecule Clinical chemistry/auto analyzer
    Steroid Clinical chemistry/auto analyzer
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Preservation Type of fixation/preservation Frozen
    None (fresh)
    Storage Storage duration 0 months
    1 month
    2 months
    3 months

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