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

High Performance Liquid Chromatography (HPLC) MS Method for Analysis of Lutein, Zeaxanthin on Primesep B by SIELC Technologies

Separation type: Liquid Chromatography Mixed-mode SIELC Technologies

HPLC Method for Analysis of Lutein, Zeaxanthin on Primesep B Column


 High Performance Liquid Chromatography (HPLC) Method for Analysis of Lutein, Zeaxanthin

Lutein is an oxygenated carotenoid, or xanthophyll, with the chemical formula C₄₀H₅₆O₂. It has a chain of conjugated double bonds and hydroxyl groups at each end, which give it strong antioxidant properties. Lutein’s ability to absorb blue light helps protect the eyes from oxidative stress. In plants, lutein stabilizes chlorophyll and protects cells from light-induced damage.

Zeaxanthin is also a xanthophyll carotenoid, sharing the same chemical formula as lutein, C₄₀H₅₆O₂, but with a different arrangement of double bonds. This structural variation allows zeaxanthin to be particularly effective in protecting the central macula of the retina. It filters harmful blue light and shields the eyes from oxidative damage, similar to lutein.

Lutein, Zeaxanthin can be retained, separated and analyzed using a Primesep B mixed-mode stationary phase column. The analysis employs an isocratic method with a simple mobile phase comprising water, acetonitrile (MeCN), and formic acid as a buffer. This method allows for detection using Vis 450 nm.

ColumnPrimesep B, 4.6 x 150 mm, 5 µm, 100 A
Mobile PhaseMeCN – 85%
BufferFormic Acid -0.1%
Flow Rate1.0 ml/min
DetectionVis 450 nm, ESI MS SIM 551 +, 568+
* LOD was determined for this combination of instrument, method, and analyte, and it can vary from one laboratory to another even when the same general type of analysis is being performed.

Class of Compounds
xanthophylls
Analyzing CompoundsLutein, Zeaxanthin

Application Column

Primesep B

Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 5 µm
Pore Size: 100 A

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Application Analytes:
Lutein
Zeaxanthin
Application Detection:
UV Detection
LC MS Detection
SIELC Technologies usually develops more than one method for each compound. Therefore, this particular method may not be the best available method from our portfolio for your specific application. Before you decide to implement this method in your research, please send us an email to research@sielc.com so we can ensure you get optimal results for your compound/s of interest.