Beta-carotene determined in serum by liquid chromatography with an internal standard

Abstract

We describe a procedure for quantitative determination of beta-carotene in human serum. The 0.1-mL serum sample is precipitated with ethanol containing the internal standard, dimethyl-beta-carotene, then extracted with hexane. This extract is injected onto a reversed-phase, 'high-performance' liquid-chromatography column, and the carotenes are resolved and eluted with an acetonitrile/methylene chloride isocratic solvent system. They are quantified from the peak-height ratios of their absorbance at 450 nm. About 14 min is required for each chromatogram. The procedure has excellent precision and is appropriate for routine use in analysis of large numbers of samples. The method should be particularly useful for clinical studies on the relationship of serum beta-carotene and cancer incidence in human populations.