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Related Experiment Videos

Candidate reference method for determining creatinine in serum: method development and interlaboratory validation.

T G Rosano1, R T Ambrose, A H Wu

  • 1Department of Pathology and Laboratory Medicine, Albany Medical Center, NY 12208.

Clinical Chemistry
|November 1, 1990
PubMed
Summary
This summary is machine-generated.

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A new high-performance liquid chromatography (HPLC) method accurately measures serum creatinine. This validated isocratic HPLC method offers a reliable alternative to existing assays for clinical diagnostics.

Area of Science:

  • Clinical Chemistry
  • Analytical Chemistry
  • Biochemistry

Background:

  • Accurate serum creatinine determination is crucial for diagnosing kidney function and related diseases.
  • Existing methods like alkaline picrate and enzymatic assays have known limitations, including potential positive bias.

Purpose of the Study:

  • To develop and validate a high-performance liquid chromatography (HPLC) method for accurate serum creatinine measurement.
  • To assess the performance characteristics and interlaboratory transferability of the proposed isocratic HPLC method.

Main Methods:

  • Serum protein precipitation followed by isocratic ion-exchange chromatography with UV detection (234 nm).
  • Validation using certified reference materials (National Institute of Standards and Technology) and comparison with isotope-dilution mass spectrometry.

Related Experiment Videos

  • Comparison with step-gradient HPLC, alkaline picrate, and enzymatic methods using patient samples.

    • Main Results:

    • The method demonstrated linearity up to 1770 µmol/L, a detection limit of 22 µmol/L, 101% average recovery, and coefficients of variation (CV) from 3% to 11%.
    • Isocratic HPLC results closely agreed with step-gradient HPLC and showed good correlation with definitive mass spectrometry measurements.
    • Confirmed positive bias of the alkaline picrate method and demonstrated interlaboratory transferability across different instruments and columns.

    Conclusions:

    • The developed isocratic HPLC method is accurate, precise, and reliable for serum creatinine determination.
    • This method serves as a strong candidate Reference Method for routine clinical laboratory use.
    • It offers advantages over traditional methods, particularly in avoiding the positive bias associated with alkaline picrate assays.