Analysis of Water-soluble Vitamins

Importance of Topic

Water-soluble vitamins play vital roles in biochemical pathways and human health. Accurate analysis is critical for nutritional quality control in food products, dietary supplements, and pharmaceutical formulations.

Objectives and Study Overview

This application demonstrates a fast, reliable reversed-phase HPLC method for simultaneous determination of ten water-soluble vitamins, leveraging Shim-pack GIST C18 stationary phase to achieve high resolution and throughput.

Methodology and Instrumentation

The separation uses a gradient from aqueous 10 mM KH₂PO₄ (pH 2.5, adjusted with H₃PO₄) containing 5 mM ion-pairing reagent to acetonitrile. The program transitions from 98/2 to 88/12 (A/B) over 13 minutes at 0.6 mL/min. Injection volume is 2 µL, column temperature 30 °C, and detection at 210 nm.

Used Instrumentation

• Shim-pack GIST C18 column (100 mm × 3.0 mm I.D., 3 µm)
• UV detector set at 210 nm
• High-performance liquid chromatography system with quaternary pump, autosampler, and column oven

Main Results and Discussion

All ten analytes—ascorbic acid, nicotinic acid, nicotinamide, pantothenic acid, pyridoxal, pyridoxine, pyridoxamine, thiamine, folic acid, and riboflavin—were baseline separated within 13 minutes. Chromatographic peaks exhibited good symmetry and minimal tailing, demonstrating the efficacy of the chosen gradient and ion-pairing conditions.

Benefits and Practical Applications

This method enables high-throughput, precise quantification of water-soluble vitamins, ideal for quality assurance in food analysis, supplement manufacturing, and clinical research settings.

Future Trends and Opportunities

Future developments may include coupling with mass spectrometry for enhanced sensitivity and specificity, implementation of UHPLC columns for faster analysis and reduced solvent consumption, and integration into automated platforms for streamlined laboratory workflows.

Conclusion

The presented reversed-phase HPLC approach using Shim-pack GIST C18 offers a robust, rapid solution for comprehensive water-soluble vitamin profiling, meeting the demands of modern analytical laboratories.