6533b81ffe1ef96bd1277c36
RESEARCH PRODUCT
Development and validation of a fast and sensitive chromatographic assay for all-trans-retinol and tocopherols in human serum and plasma using liquid-liquid extraction.
Taibi GC. M. A. Nicotrasubject
AnalyteChromatographyClinical BiochemistryExtraction (chemistry)RetinolReproducibility of ResultsTocopherolsLiterCell BiologyGeneral MedicineBiochemistryHigh-performance liquid chromatographySensitivity and SpecificityAnalytical Chemistrychemistry.chemical_compoundSpectrometry FluorescencechemistryLiquid–liquid extractionHumansSample preparationAll trans retinolVitamin AChromatography High Pressure Liquiddescription
A sensitive HPLC assay for all-trans-retinol, alpha-tocopherol, and gamma-tocopherols in human serum and plasma is reported. Sample preparation is performed in one step and involves precipitation of proteins and extraction of lipids with two volumes of an ethanol-chloroform mixture (3:1, v/v) without I.S. addition. After removal of the precipitated protein, 20 microl aliquots of the supernatant (equivalent to 6.7 microl of serum or plasma) were injected into the HPLC system and analyzed using fluorometric detection. RP-HPLC was performed using a C(18) S3 ODS2 column with a methanol-water step gradient (97:3 to 100) at 1.0 ml/min. The quantification limit expressed as nanograms of analyte per milliliter of serum or plasma was approximately 30 ng for all-trans-retinol, 300 ng for alpha-tocopherol and 250 ng for gamma- and delta-tocopherol. The method was validated and applied to human serum and plasma from a total of 120 subjects. This procedure requires a small volume of serum or plasma and can therefore be a valuable tool for measuring low concentrations of these vitamins in preterm infants with sensitivity, precision and accuracy.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2002-11-01 | Journal of chromatography. B, Analytical technologies in the biomedical and life sciences |