Analysis of Lutein and Zeaxanthin

Importance of the Topic

Lutein and zeaxanthin are dietary xanthophylls with significant roles in human health, particularly in eye function and antioxidant protection. Precise quantification of these carotenoids is essential for quality control in food products, nutritional research and regulatory compliance.

Objectives and Study Overview

This application note presents a rapid, high-resolution reversed-phase liquid chromatography method for the simultaneous separation and quantification of lutein and zeaxanthin in food and supplement matrices. The study focuses on optimizing chromatographic conditions to achieve baseline separation within a short run time.

Methodology

The procedure uses a Shim-pack XR-ODS column (75 mm × 3.0 mm, 2.2 µm) with a binary gradient:

• Mobile phase A: Methanol/Tetrahydrofuran/Water (45/30/25, v/v/v)
• Mobile phase B: Tetrahydrofuran
• Gradient program: 0% B (0.00–4.50 min) → 100% B (4.51–5.50 min) → 0% B (5.51–7.50 min)
• Flow rate: 0.8 mL/min
• Column temperature: 50 °C
• Injection volume: 2 µL
• Detection: UV-VIS at 450 nm

Used Instrumentation

• Shimadzu LC system equipped with Shim-pack XR-ODS column (P/N 228-41606-93)
• UV-VIS detector set to 450 nm

Main Results and Discussion

The method achieved clear baseline separation of zeaxanthin and lutein within a 7.5 min run. Sharp peak shapes and minimal tailing confirm the effectiveness of the small particle size column and optimized gradient. The fast cycle time enhances throughput without sacrificing resolution.

Benefits and Practical Applications

• High sensitivity and selectivity for routine analysis of carotenoids in food, supplements and biologically relevant samples
• Short analysis time supports high sample throughput in quality control laboratories
• Reliable quantification aids in compliance with labeling regulations and research on nutrient bioavailability

Future Trends and Applications

Extension of this method to additional carotenoids and adaptation for coupling with mass spectrometry could further improve identification capabilities. Integration into automated QA/QC workflows and its application in nutraceutical development and clinical studies are promising avenues.

Conclusion

The described reversed-phase LC method on Shim-pack XR-ODS provides a robust, fast and sensitive solution for simultaneous analysis of lutein and zeaxanthin. It meets the demand for efficient high-throughput testing in food science and nutritional research.

References

• Shimadzu Corporation. Application Note ERAS-1000-0523: Analysis of Lutein and Zeaxanthin, First Edition, Sep. 2023.