HPLC Method for Analysis of Lutein and Zeaxanthin in Supplements on Zodiac HST B Column

Lutein is an oxygenated carotenoid, classified as a xanthophyll, with the chemical formula C₄₀H₅₆O₂. It features a chain of conjugated double bonds and terminal hydroxyl groups, contributing to its potent antioxidant properties. Lutein is particularly effective at absorbing blue light, offering protection against oxidative stress in the eyes. In plants, it plays a vital role in stabilizing chlorophyll and safeguarding cells from light-induced damage.

Zeaxanthin, also a xanthophyll carotenoid with the same chemical formula (C₄₀H₅₆O₂), differs in its double bond configuration. This structural difference enhances its ability to protect the central macula of the retina. Like lutein, it filters harmful blue light and helps prevent oxidative damage to eye tissues.

Analytical Method:

Lutein and zeaxanthin can be effectively retained, separated, and analyzed using the Zodiac HST B mixed-mode stationary phase column. The method utilizes a simple isocratic mobile phase consisting of water, acetonitrile (MeCN), and sulfuric acid as the buffer. Detection is carried out at 450 nm using visible light (Vis), making this approach suitable for the analysis of xanthophyll carotenoids.