Ultrafast Analysis of Water-Soluble Vitamins

Importance of the Topic

Water-soluble vitamins play crucial roles in metabolism, cellular processes and public health. Rapid and reliable quantification of B-complex vitamins such as thiamine, riboflavin and niacin derivatives is essential in nutritional analysis, quality control of dietary supplements and functional food development. Advanced chromatographic techniques enable high throughput and precise determination of multiple vitamins in complex matrices.

Aims and Study Overview

This application demonstrates an ultrafast gradient elution method using a core-shell C18 column coupled with Nexera UHPLC for simultaneous separation of six water-soluble vitamins. Two workflows are presented: analysis of standard mixtures to assess repeatability and linearity, and practical testing on commercial nutrition drink samples to illustrate applicability in real matrices.

Methodology and Instrumentation

Key analytical parameters and instrumentation:

• UHPLC System: Shimadzu Nexera with high-accuracy gradient capability
• Column: Phenomenex Kinetex C18, core-shell, 100 × 4.6 mm, 2.6 µm particles
• Mobile Phases: A = 20 mmol/L phosphate buffer (pH 2.5) with 2 mmol/L sodium 1-hexanesulfonate; B = A/ACN (2/3)
• Gradient: 5 % B at 0 min→23 % B at 1 min→100 % B at 2–2.5 min
• Flow Rate: 2.5 mL/min (180 µL mixer volume)
• Column Temperature: 40 °C
• Injection Volume: 5 µL for standards; 2–5 µL for sample tests
• Detection: UV at 275 nm, semi-micro flow cell
• Operating Pressure: Up to 48 MPa

Main Results and Discussion

Standard mixture analysis of six vitamins (niacin, nicotinamide, pyridoxine, riboflavin phosphate, thiamine, riboflavin) yielded sharp peaks within 2.5 min. Repeatability (n=6) showed:

• Retention time RSD ≤ 0.05 %
• Peak area RSD ≤ 0.10 %

Gradient precision and injection repeatability underscore the system’s robustness. Analysis of two commercial nutrition drinks confirmed clear separation of all analytes within the same runtime, demonstrating minimal matrix interference and reliable quantification.

Benefits and Practical Applications

The ultrafast method offers:

• High throughput: complete separation in under 3 minutes
• Excellent precision: low RSD in retention and peak area
• Broad applicability: suitable for standards and complex beverages
• Reduced solvent consumption: short gradient and core-shell efficiency

These features support routine QA/QC in supplement manufacturing, regulatory compliance and research laboratories.

Future Trends and Application Possibilities

Emerging directions include:

• Integration with mass spectrometry for enhanced sensitivity and specificity
• Miniaturization and microfluidic platforms for on-site testing
• Automated sample preparation to further increase throughput
• Expansion to other vitamin classes and water-soluble biomarkers

Adoption of faster, greener chromatography aligns with sustainability goals and the demand for real-time monitoring in the food and pharmaceutical sectors.

Conclusion

The presented ultrafast gradient LC method using a core-shell C18 column and Nexera system achieves precise, reproducible separation of six water-soluble vitamins in under 3 minutes. Its high throughput, accuracy and adaptability make it a valuable tool for nutritional analysis, quality control and research applications.

Reference

Shimadzu Corporation. Ultrafast Analysis of Water-Soluble Vitamins. Nexera Application Data Sheet No.16. Issued Mar 2011.