Fluorometric determination of retinol in human tear fluid using high-performance liquid chromatography
article
A fast and sensitive high-performance liquid chromatographic (HPLC) method for the determination of vitamin A (all-trans retinol) in 50 ??l human tear fluid is described. After deproteinization with ethanol and extraction with n-hexane, retinol is separated from the extract on a straight-phase HPLC column and detected fluorometrically. Vitamin A can be determined in concentrations as low as 0.4 ng/ml. A single analysis can be completed in 12 min while the analysis of a series of 60 samples takes about 8 h. The within-assay and between-assay coefficients of variation were 3.1 and 4.2% resp. The between-assay analytical recovery of retinol added to tear fluid was 97.4 ?? 6.0% (mean ?? SD). Retinol was determined in tear fluid of nine adult volunteers. The amounts observed ranged from <0.4-10.6 ng/ml. Chemicals/CAS: retinol, 68-26-8, 82445-97-4; Vitamin A, 11103-57-4
Topics
RetinolDrug analysisDrug determinationDrug tissue levelFluorometryHigh performance liquid chromatographyHumanHuman cellMethodologyNormal humanTearVisual systemChromatography, High Pressure LiquidDrug StabilityEvaluation StudiesFluorometryHumanSupport, Non-U.S. Gov'tTearsVitamin ABiologyBiochemistryMedicineGeneeskunde
TNO Identifier
230190
ISSN
02713683
Source
Current Eye Research, 5(11), pp. 841-845.
Pages
841-845
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