Use of a 13C tracer to investigate lutein as a ligand for plasma transthyretin in humans.

The selective accumulation of lutein in the macula of the human retina is likely to be mediated by specific transport and/or binding proteins. Our objective was to determine whether transthyretin (TTR) is a plasma transport protein for lutein. We used a biosynthetic 13C-lutein tracer and GC-combustion interfaced-isotope ratio MS to gain the requisite sensitivity to detect the minute amounts of lutein expected as a physiological ligand for TTR. Subjects (n = 4) each ingested 1 mg of 13C-lutein daily for 3 d and donated blood 24 h after the final dose. For three subjects, the plasma TTR-retinol-binding protein (RBP) complex was partially purified by anion-exchange (diethylaminoethyl, DEAE) chromatography and then dissociated by hydrophobic-interaction chromatography to yield the TTR component. For subject 4, the initial DEAE purification step was omitted and total plasma TTR (RBP-bound and free) was isolated by hydrophobic-interaction chromatography. In each case, the crude TTR fractions were then purified to homogeneity by RBP-Sepharose affinity chromatography. Pure TTR was extracted with chloroform, and unlabeled lutein was added to the extract as a carrier. The mean 13C/12C ratio (expressed in delta notation, delta13C) of the lutein fraction isolated from the plasma TTR extracts of the four subjects was -30.53 +/- 3.29 per thousand. The delta13C value of the unlabeled lutein carrier was -30.97 +/- 0.27per thousand. Thus, no 13C enrichment was detected in association with TTR. We conclude that lutein is not associated with TTR in human plasma after being ingested in physiological amounts.