Chromatography for Foods and Beverages - Vitamin and Antioxidant Applications Notebook

Importance of the Topic

Modern food, beverage and supplement quality control relies on accurate, sensitive and high-throughput analysis of vitamins and antioxidants. These compounds, essential for human health and nutrition, vary widely in chemical properties, from highly polar water-soluble B and C vitamins to nonpolar fat-soluble A, D, E and K forms and nonessential dietary antioxidants. Reliable analytical methods ensure label compliance, detect degradation products and contaminants, and support nutritional research and regulatory requirements.

Study Objectives and Overview

This collection of proven analytical applications demonstrates robust chromatographic and detection techniques for:

• Individual and simultaneous measurement of water- and fat-soluble vitamins
• Quantification of vitamin mixtures in tablets, beverages, feeds and biological samples
• Profiling of non-essential antioxidants such as carotenoid isomers, CoQ10, lipoic acid and carnitine derivatives

Examples cover high-performance liquid chromatography (HPLC), ultra-high performance liquid chromatography (UHPLC), ion chromatography (IC), mass spectrometry (MS) coupling and advanced detection including diode array (DAD), charged aerosol (CAD), electrochemical (CoulArray) and fluorescence detectors.

Methodology and Instrumentation

Common workflows employ reversed-phase UHPLC/HPLC with gradients tailored to compound polarity. Polar-embedded and phenyl columns permit retention of both polar and nonpolar vitamins in a single run. Key methodologies include:

• Online solid-phase extraction (SPE) coupled to HPLC-UV for trace vitamin B12 in beverages
• HILIC and normal-phase separations for lipid-soluble vitamins in oil formulations
• Simultaneous water- and fat-soluble vitamin assays using amide-embedded columns under dual detection wavelengths
• Ion chromatography for inorganic anion/cation vitamins like ascorbate and pantothenate without reagent addition
• UHPLC-MS for high-sensitivity confirmation and quantitation
• Coulometric array detection for profiling of fat-soluble antioxidants

Instrumentation frequently used: Thermo Scientific™ UltiMate™ 3000 UHPLC+ systems, Dionex™ Ion Chromatography systems (ICS-5000+, RFIC), MSQ Plus single quadrupole MS, Dionex Corona Veo™ CAD, CoulArray™ electrochemical detectors, VWD, DAD and fluorescence modules, ASE™ accelerated solvent extractor and AutoTrace™ SPE platforms.

Main Results and Discussion

Applications achieve:

• Baseline-resolved separation of eight water-soluble vitamins in 5 minutes using UHPLC amide columns
• Complete fat-soluble vitamin profiles in 20 minutes by RP-HPLC-CAD with detection limits in the low nanogram range
• Quantitative vitamin B12 analysis in functional beverages down to sub-nanogram per milliliter levels via online SPE–HPLC-UV
• Simultaneous assay of multivitamin tablets and infant formulas with recoveries within 95–105%
• Carotenoid isomer resolution in tomatoes, plasma and food matrices by C30 columns with DAD
• Global lipid antioxidant profiling using CoulArray with selective potentials for individual compounds

Advantages and Practical Applications

These methods offer:

• High throughput and reduced solvent consumption through sub-2 μm UHPLC columns
• Enhanced selectivity for co-eluting isomers via HILIC, phenyl and C30 chemistries
• Versatile detection adapting to UV-active and non-UV-active analytes using CAD and electrochemical sensors
• Regulatory compliance with built-in instrument wellness monitoring and automated qualification
• Minimal sample preparation via online SPE and accelerated extraction
• Seamless integration of IC and LC systems with MS for confirmation and trace analysis

Future Trends and Applications

Emerging directions include:

• Automated lab-to-process analytical pipelines for real-time quality control in manufacturing
• Further miniaturization and increased multiplexing of detectors for ultrafast screening
• Advanced column technologies combining mixed-mode retention for comprehensive multi-analyte assays
• Machine learning-driven method development and data analysis in chromatography data systems
• Integration of high-resolution MS and ion mobility for isomer differentiation in complex matrices

Conclusion

State-of-the-art chromatography and detection platforms enable accurate, sensitive and efficient analysis of a broad spectrum of vitamins and antioxidants. The described methods support nutritional labeling, research, quality assurance and process control across foods, beverages, supplements and biological matrices.

References

No specific references were provided in the original source document.