Simultaneous Determination of Water-soluble Vitamins

Importance of the Topic

Water-soluble vitamins play essential roles in cellular metabolism, neurological function, and overall health. Accurate and efficient analysis of these vitamins in food, biological samples, and pharmaceutical formulations is critical for quality control and regulatory compliance. A robust chromatographic method capable of simultaneously resolving multiple vitamins streamlines laboratory workflows and reduces analysis time and costs.

Objectives and Overview of the Study

The primary goal was to develop and validate a reversed-phase liquid chromatography method for the concurrent quantification of eight water-soluble vitamins. The study focused on optimizing separation conditions, achieving baseline resolution, and demonstrating method reproducibility using Shimadzu Shim-pack MAqC-ODS I column technology.

Methodology and Instrumentation

The method was carried out on a Shimadzu LC system configured with a Shim-pack MAqC-ODS I column (150 mm length, 4.6 mm i.d., 5 μm particle size). Mobile phases consisted of a 10 mmol/L sodium phosphate buffer at pH 2.6 and acetonitrile. A gradient program increased organic content from 1 to 50 over 10 minutes, returning to initial conditions by 10.01 minutes, with a total run time of 15 minutes. Flow rate was set at 1.2 mL/min, column temperature at 40 °C, and detection performed at 210 nm using a UV detector. Sample injection volume was 10 µL.

Main Results and Discussion

The optimized gradient enabled clear separation of thiamine, niacin, nicotinamide, pyridoxine, calcium pantothenate, folic acid, riboflavin, and biotin in a single run. Retention times were well spaced, and peak shapes were sharp and symmetric. The method demonstrated good repeatability with relative standard deviations below 2 for retention times and peak areas. Sensitivity was sufficient for quantifying vitamins at concentrations around 20 mg/L.

Benefits and Practical Applications

• Concurrent analysis reduces total run time and solvent consumption.
• High resolution minimizes interference between closely eluting compounds.
• Method can be applied in food fortification assessment, pharmaceutical quality control, and nutritional analysis.

Future Trends and Applications

Advances in column technology and faster gradient systems may further shorten analysis time. Coupling with mass spectrometry could enhance sensitivity and specificity for trace-level detection. Automation and high-throughput platforms will expand applicability in routine industrial settings.

Conclusion

The described LC method utilizing a Shim-pack MAqC-ODS I column offers a rapid, reliable, and reproducible approach for simultaneous determination of key water-soluble vitamins. Its implementation supports efficient quality control in diverse analytical laboratories.

Reference

• Application News L459 JP ENG Shimadzu Corporation First Edition Dec 2021