Analysis of Water-Soluble Vitamins with Multi-Mode ODS Column Using LCMS-2020

Importance of the Topic

Water-soluble vitamins play essential roles in human health and nutrition. Their analysis in functional foods and supplements is critical to ensure compliance with dietary guidelines and consumer safety. Due to their high polarity, these compounds exhibit weak retention on conventional reversed-phase columns, often requiring ion-pair reagents that can compromise MS sensitivity.

Study Objectives and Overview

This work demonstrates the simultaneous analysis of eight water-soluble vitamins using a cation-anion exchange multimode ODS column (Scherzo SM-C18) coupled to the LCMS-2020 mass spectrometer. Objectives included achieving adequate retention, separation, and quantitation with high sensitivity and linearity.

Methodology and Instrumentation

The analysis employed a gradient of formic acid/ammonium formate buffer (mobile phase A) and acetonitrile (mobile phase B) on a Scherzo SM-C18 column (150×2.0 mm, 3 μm) at 40 °C. Key analytical conditions:

• Flow rate: 0.2 mL/min; injection volume: 2 μL
• Gradient: 0→55 % B over 10 min, return to 0 % B by 10.01 min, hold to 20 min
• Mass spectrometry: LCMS-2020 with ESI in positive and negative modes
• Probe voltages: +4.5 kV (positive), −3.5 kV (negative)
• Nebulizing gas: 1.5 L/min; drying gas: 10 L/min
• DL temperature: 250 °C; block heater: 450 °C

Key Results and Discussion

Baseline separation of thiamin, pyridoxine, nicotinic acid, nicotinamide, pantothenic acid, cyanocobalamin, riboflavin, and folic acid was achieved within 12.5 min. Mass spectra showed clear [M+H]+ or [M–H]– ions. Calibration curves exhibited excellent linearity (R2>0.99) across relevant concentration ranges (e.g., 1–1000 ng/mL for thiamin). Application to cereal extract samples using ultrasonic extraction followed by centrifugation demonstrated quantitation with minimal matrix interference. Cyanocobalamin was below detection in the tested sample.

Benefits and Practical Applications

This method offers:

• High sensitivity and selectivity without ion-pair reagents
• Rapid, simultaneous quantitation of multiple vitamins
• Minimal sample preparation and robust performance for food analysis

Future Trends and Potential Applications

Ongoing advances in multimode stationary phases and MS detector designs will further enhance throughput and sensitivity. Integration with high-resolution MS and automated sample handling will expand applications to clinical diagnostics and metabolomics.

Conclusion

The Scherzo SM-C18 column combined with LCMS-2020 provides an effective platform for the rapid, sensitive analysis of water-soluble vitamins in complex matrices. The approach meets the needs of quality control in the food and supplement industries.