Nonaqueous reverse phase liquid chromatographic system for separation and quantitation of provitamins A.
A reverse phase liquid chromatographic (LC) system has been developed for separating the main naturally occurring carotenoids that have provitamin A activity. The system produces baseline separation of beta-carotene, alpha-carotene, and beta-cryptoxanthin (beta, beta-carotene-3-ol, 472-70-8) from biologically inactive zeinoxanthin (beta, epsilon-carotene-3-ol, 24480-38-4) and from a pigment believed to be alpha-cryptoxanthin (beta, epsilon-carotene-3'-ol). Some cis-isomers are also separated. These separations are obtained on a C-18 column, isocratically, with methanol-chloroform eluant. For quantitation, peak areas from detection at 475 nm are compared with that of an internal standard, 1-(phenylazo)-2-naphthalenol (842-07-9), which elutes prior to the provitamins. Provitamin amounts are calculated from absorbance ratios. Prior to LC, esters are saponified, and interfering pigments are removed from ester-free extracts by adsorption on magnesia.
D002338:Carotenoids / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D007202:Indicators and Reagents; D013053:Spectrophotometry / Q000379:methods; D013329:Structure-Activity Relationship; D014801:Vitamin A / Q000302:isolation & purification*
D007202:Indicators and Reagents; D014801:Vitamin A; D002338:Carotenoids