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Validation of an In-House-Developed GC-MS Method for 5α-Cholestanol According to ISO 15189:2012 Requirements
1Dokuz Eylül University Faculty of Medicine, Department of Biochemistry, Izmir, Turkey.
Objective:
The aim of this study was to validate a gas chromatography-mass spectrometry (GC-MS) method for the measurement of 5α-cholestanol in the clinical laboratory in agreement with ISO 15189:2012.
Materials And Methods:
The GC-MS performance was evaluated and proficiency testing data were used to estimate the measurement uncertainty of the method considering the recommendations of international guidelines.
Results:
The calibration curves were linear from 6 to 50 μmol/L, with r2 >.99. The limit of detection and limit of quantitation were determined to be 0.36 and 2.58 μmol/L, respectively. The bias ranged from -18.9% to 15.2% for 6.5, 18.3, and 66 μmol/L. The intra- and interassay reproducibility was <20% at the various concentrations studied. The expanded uncertainty was determined to be 50.9%.
Conclusion:
The GC-MS method for the measurement of 5α-cholestanol has proved to have acceptable analytical performance for use in the clinical laboratory.
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