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

Lutein is an oxygenated carotenoid (xanthophyll) with the molecular formula C₄₀H₅₆O₂. Its structure features a series of conjugated double bonds and hydroxyl groups at both ends, which contribute to its potent antioxidant activity. Lutein is known for its ability to absorb blue light, offering protection against oxidative stress, particularly in the eyes. In plants, it plays a crucial role in stabilizing chlorophyll and shielding cells from light-induced damage.

Zeaxanthin is another xanthophyll carotenoid that shares the same chemical formula (C₄₀H₅₆O₂) as lutein but differs in the arrangement of its double bonds. This structural distinction makes zeaxanthin especially effective in safeguarding the central macula of the retina. Like lutein, it filters harmful blue light and protects ocular tissues from oxidative stress.

Analytical Method:

Lutein and zeaxanthin can be efficiently retained, separated, and analyzed using a Zodiac HST B mixed-mode stationary phase column. The method utilizes a simple isocratic mobile phase consisting of water, acetonitrile (MeCN), and formic acid as the buffer. Detection is performed using visible light at 450 nm, making this a robust and effective approach for carotenoid analysis.