Determination of B-vitamins in Energy Drinks by CE/MS/MS

Importance of the topic

Energy drinks often contain B-vitamins essential for human health. Accurate quantification is critical to ensure label compliance, assess nutrient stability, and safeguard consumer health. Traditional assays may lack speed or multiplexing capability, motivating advanced analytical approaches.

Objectives and overview

This study aims to develop and validate a fast, sensitive capillary electrophoresis-tandem mass spectrometry (CE/MS/MS) method for simultaneous determination of seven B-vitamins in energy drinks: B1, B2, B3a, B3b, B5, B6, and B12, using cetirizine as an internal standard.

Methodology

• Background electrolyte: 50 mM formic acid, pH 2.5.
• Capillary: fused silica, 50 µm i.d., 60 cm total length.
• Voltage: 29 kV at 25 °C; hydrodynamic injection: 5 s at 50 mbar.
• Sample preparation: degassing in ultrasonic bath, 0.2 µm PVDF filtration, 1:10 dilution.

Used instrumentation

• Capillary electrophoresis: Agilent 7100 CE system.
• Mass spectrometry: Agilent 6430 triple quadrupole with ESI positive mode; MRM acquisition.
• Sheath liquid: 0.1% formic acid in methanol (50:50), 5 µL/min.
• ESI settings: capillary voltage 2.5 kV, drying gas 7 L/min at 350 °C, nebulizer 9 psi.

Main results and discussion

Under optimized conditions, all seven vitamins and the internal standard were separated in under 8 minutes with excellent peak shapes. Calibration was linear with R2 > 0.995 across relevant concentration ranges. Limits of detection ranged from 0.01 to 0.19 ppm, and limits of quantification from 0.04 to 0.63 ppm. Injection volumes were approximately 4.5 nL. Precision (RSD) was below 6.6%. Application to 11 commercial samples yielded results consistent with label claims; minor discrepancies in B1 and B3 levels were attributed to natural guarana extract.

Benefits and practical applications

The method is rapid, consumes minimal sample and reagents, and generates little waste, making it suitable for high-throughput quality control of fortified beverages and other food products.

Future trends and potential applications

• Coupling CE with high-resolution mass spectrometry for broader analyte coverage.
• Microchip and nano-CE platforms for further reduction in sample and solvent needs.
• Automated workflows for routine quality assurance in industry.
• Development of portable CE-MS devices for on-site analysis.

Conclusion

The developed CE/MS/MS approach offers a robust, sensitive, and environmentally friendly solution for simultaneous determination of B-vitamins in energy drinks, fulfilling the demands of modern analytical laboratories.

References

• de Leenheer AP, Lambert WE, Van Bocxlaer JF, editors. Modern Chromatographic Analysis of Vitamins. 3rd ed. Marcel Dekker; 2000.
• Simó C, Barbas C, Cifuentes A. Capillary electrophoresis-mass spectrometry in food analysis. Electrophoresis. 2005;26:1306-1318.
• Wang X, et al. Recent advances in vitamins analysis by capillary electrophoresis. Journal of Pharmaceutical and Biomedical Analysis. 2018;147:278-287.
• Fotsing L, et al. Determination of six water-soluble vitamins in a pharmaceutical formulation by capillary electrophoresis. Journal of Pharmaceutical and Biomedical Analysis. 1997;15:1113-1124.