Rapid and sensitive UHPLC screening for water soluble vitamins in sports beverages

Importance of the topic

Water-soluble vitamins are critical for human health and must be accurately quantified in fortified beverages to ensure safety and label compliance.

Objectives and study overview

The study demonstrates a rapid UHPLC method using Thermo Scientific Acclaim PolarAdvantage II column and Vanquish Flex Binary system for simultaneous analysis of seven water-soluble vitamins in sports drinks with high precision, resolution, and recovery.

Methodology and instrumentation

• Instrumentation: Vanquish Flex Binary UHPLC system with Binary Pump F, System Base, Split Sampler FT, Column Compartment H, Active Pre-heater, Diode Array Detector HL, LightPipe Flow Cell (2 μL, 10 mm), Virtuoso vial identification system.
• Column: Acclaim PolarAdvantage II PA2 (100 mm × 3 mm, 2.2 μm).
• Mobile phases: 0.015% formic acid in water (A) and 20:80 methanol/acetonitrile (B); gradient from 100% A to 45% B in 6 minutes, flow rate 0.5 mL/min, column temperature 50 °C.
• Detection: UV at 210 nm.
• Sample preparation: beverages degassed, diluted 1:1 with mobile phase A, filtered, spiked with mixed vitamin standard; calibration standards prepared in mobile phase A.

Key results and discussion

• Separation: Baseline resolution of seven vitamins achieved in approximately 6 minutes.
• Precision: Retention time RSD ≤0.27% and peak area RSD ≤1.82% for all compounds.
• Linearity: Calibration curves with R2 between 0.9953 and 0.9983 over relevant concentration ranges.
• Recovery: Spiked recoveries ≥80% in three sports drink matrices, indicating minimal matrix interference.
• Quantification: Measured vitamin concentrations closely matched product label values; vitamin B12 was below the lower limit of quantification in all samples.

Benefits and practical applications

The method offers rapid, sensitive, and reproducible analysis for quality control of vitamin-fortified beverages, supporting regulatory compliance and product development in the food and beverage industry.

Future trends and applications

Advances may include coupling UHPLC to mass spectrometry for enhanced selectivity, high-throughput automation, expansion to diverse matrices, method miniaturization, and integration into online process monitoring for real-time nutrient profiling.

Conclusion

The combination of the Acclaim PolarAdvantage II column and Vanquish Flex Binary system provides a fast and reliable UHPLC method for high-throughput screening of water-soluble vitamins in sports beverages with excellent performance metrics.

Instrumentation

• Thermo Scientific Vanquish Flex Binary UHPLC system (Binary Pump F, System Base, Split Sampler FT, Column Compartment H, Active Pre-heater, Diode Array Detector HL)
• Thermo Scientific Acclaim PolarAdvantage II (PA2) column, 100 mm × 3 mm, 2.2 μm
• LightPipe Flow Cell (2 μL, 10 mm)
• Virtuoso vial identification system

Reference

Bardsley J. Rapid and sensitive UHPLC screening for water soluble vitamins in sports beverages. Thermo Fisher Scientific Application Note AN21671, 2017.