Journal
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
Volume 877, Issue 11-12, Pages 1209-1215Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jchromb.2009.02.063
Keywords
Serum; Retinol; alpha-Tocopherol; HPLC; Validation; Uncertainty of measurement; Biological variability; Quality specifications
Funding
- Istituto Superiore di Sanita [1033/2033/RI 2000-2002]
- Italian Ministry of Health Project [Alz7 2001-2005]
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An isocratic RP-HPLC method for the determination of retinol and a-tocopherol in serum, with fluorescence and UV/VIS detection, respectively, was developed and validated according to international guidelines. Detection (retinol 0.015 mg/L, alpha-tocopherol 0.29 mg/L) and quantification (retinol 0.05 mg/L, a-tocopherol 0.95 mg/L) limits were determined. Repeatability was < 3.3% and < 2.9% and intermediate precision was < 4.6% and < 3.2%, for retinol and a-tocopherol, respectively. Certified reference materials were utilised to assess bias and guarantee traceability to SI units. Expanded uncertainties (retinol 8.9%: a-tocopherol 7.9%), estimated according to the EURACHEM/CITAC guide from method validation data, satisfied fit-for-purpose requirements based on biological variability. (C) 2009 Elsevier B.V. All rights reserved.
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