Simultaneous Analysis of Water- and Fat-Soluble Vitamins in Beverages Using an ODS-Modified Metal-Doped Column

Importance of the topic

Vitamins are essential micronutrients that support metabolic processes in the human body. Simultaneous analysis of both water- and fat-soluble vitamins is critical for quality control of fortified beverages and dietary supplements. A unified method reduces analysis time and avoids complex sample preparation.

Study objectives and overview

This study aimed to develop a single high-performance liquid chromatography method with a photodiode array detector (HPLC-PDA) capable of separating ten water-soluble and nine fat-soluble vitamins in one run. The goal was to eliminate ion-pair reagents and simplify gradient elution.

Methodology and instrumentation

The method uses a Shimadzu Prominence-i integrated HPLC system with PDA detection. Separation is achieved on a Shim-pack MAqC-ODS I column (150 mm × 4.6 mm, 5 µm) and guard column Shim-pack GVP-ODS (10 mm × 4.6 mm, 4.6 µm). Mobile phases consist of 10 mmol/L phosphate buffer (pH 2.6) and acetonitrile. A gradient from 0 % to 40 % organic over 10 minutes, then to 100 % by 25 minutes, was applied at 1.2 mL/min with column temperature of 40 °C. Injection volume was 10 µL and detection wavelength set at 220 nm.

Main results and discussion

The ODS-modified metal-doped stationary phase improved retention of polar water-soluble vitamins without ion-pairing agents. All ten water-soluble and nine fat-soluble vitamins were baseline-separated in under 45 minutes. In fortified water samples, most vitamins were detected at 220 nm; retinyl palmitate required detection at 330 nm due to low concentration. Chromatograms demonstrated consistent peak shapes and reproducible retention times.

Benefits and practical applications

• Single-run analysis increases laboratory throughput
• Avoidance of ion-pair reagents simplifies method development and maintenance
• Wide applicability to quality control of fortified beverages and nutritional products

Used instrumentation

• Shimadzu Prominence-i integrated HPLC system with PDA model LC-2030 3D
• Shim-pack MAqC-ODS I column and Shim-pack GVP-ODS guard column

Future trends and potential applications

Further development may include coupling with mass spectrometry for enhanced sensitivity and selectivity. Faster gradients and smaller particle columns could reduce analysis time. The method can be extended to complex matrices such as dairy products and fortified foods.

Conclusion

The Shim-pack MAqC-ODS I column enabled robust simultaneous separation of water- and fat-soluble vitamins in a single HPLC-PDA run, offering a streamlined approach for routine quality control and research applications.

References

• Shimadzu application news No. L459 Simultaneous Analysis of Water-Soluble Vitamins by a Newly Developed Shim-pack MAqC-ODS I Column, First Edition March 2015