Selective Quantitative Determination of Water Soluble Vitamins in Various Food Matrices Using the ACQUITY UPLC H-Class System and ACQUITY QDa Mass Detector

Significance of Water-Soluble Vitamin Analysis

Food and beverage products are often fortified with water-soluble vitamins to comply with nutritional regulations and address dietary deficiencies. Accurate measurement of these vitamins is vital for consumer safety, label verification, and maintaining product consistency across diverse matrices.

Objectives and Study Overview

This application note describes a streamlined UPLC-MS approach for the simultaneous quantification of twelve water-soluble vitamins in dietary supplements, powdered beverages, and vitamin waters. By coupling an ACQUITY UPLC H-Class system with an ACQUITY QDa Mass Detector, the study aims to achieve rapid analysis, improved selectivity, and lower limits of quantification compared to conventional UV-based methods.

Instrumentation and Methodology

Sample Preparation:

• Simple dissolution of powders or tablets followed by filtration through 0.2 µm PVDF membranes
• Appropriate dilutions (up to 1:1000) to match calibration ranges

Chromatography:

• ACQUITY UPLC H-Class with HSS T3 column (2.1×100 mm, 1.8 µm)
• Mobile phases: 10 mM ammonium formate with 0.1% formic acid in water (A) and methanol (B)
• Gradient elution over 17.5 min at 0.45 mL/min, 30 °C column temperature

Detection:

• ACQUITY QDa Mass Detector in positive electrospray mode with single ion recording (SIR) channels for each vitamin
• Parallel ACQUITY PDA scanning from 210–400 nm with an analog channel at 270 nm

Main Results and Discussion

All twelve water-soluble vitamins eluted within eight minutes. Mass detection enabled accurate quantification of co-eluting pairs (e.g., nicotinamide/pyridoxine and cyanocobalamin/folic acid) by monitoring unique m/z values. Calibration showed linear responses (r2>0.99) over the studied ranges, and SIR detection achieved low-µg/L sensitivity for challenging analytes like biotin and pantothenate. Trace-level cyanocobalamin (<50 µg/L) in supplement tablets was quantified reliably, whereas UV detection alone failed at these concentrations. Repeatability tests yielded retention time RSDs below 0.6% and peak area RSDs below 15% for most vitamins.

Benefits and Practical Applications

Key advantages of the UPLC-QDa method include:

• Consolidated analysis of twelve vitamins in a single run
• Minimal sample preparation with straightforward dilutions
• Enhanced selectivity overcoming co-elution without two-dimensional chromatography
• Improved detection limits facilitating compliance with strict label requirements
• Seamless integration into existing LC workflows using standard chromatography software

Future Trends and Application Possibilities

Potential developments:

• Extension to fat-soluble vitamins and broader micronutrient panels
• High-throughput screening of complex food matrices
• Automated sample preparation for routine quality control
• Adoption of high-resolution mass spectrometry for expanded compound identification

Conclusion

The combination of ACQUITY UPLC H-Class chromatography and QDa mass detection provides a rapid, sensitive, and user-friendly platform for comprehensive water-soluble vitamin analysis in diverse food and beverage products, supporting both regulatory compliance and quality assurance.

Reference

1. Ball GFM, Editor. Bioavailability and Analysis of Vitamins in Foods. Chapman & Hall; 1998.
2. Riches E. The Rapid Simultaneous Analysis of 12 Water Soluble Vitamin Compounds. Waters Application Note 720003052en; 2009.
3. Heudi O, et al. Separation of water-soluble vitamins by reversed-phase HPLC with UV detection: Application to polyvitaminated premixes. J Chromatogr A. 2005;1070:49–56.
4. AOAC Official Method 2011.09: Vitamin B12 Determination.