LC-MS/MS Method for the Rapid Analysis of Five Artificial Sweeteners Using a Core Enhanced Technology Column

Significance of the Topic

Artificial sweeteners are widely used in food and beverage industries to address health concerns such as obesity, diabetes, and dental decay. Rapid and reliable analysis of these compounds is essential for quality control and regulatory compliance, especially for sweeteners lacking UV chromophores that challenge traditional HPLC-UV detection.

Objectives and Study Overview

This study aimed to demonstrate a fast and efficient LC-MS/MS method for simultaneous quantification of five artificial sweeteners—acesulfame potassium, saccharin, sodium cyclamate, aspartame, and sucralose—using a Thermo Scientific Accucore RP-MS column. The goal was to achieve baseline separation within a minimal run time and robust detection through MS/MS transitions.

Methodology and Instrumentation

Sample Preparation:
Sweetener standards were prepared at 200 ng/mL in LC-MS grade water.

Chromatographic Conditions:

• Column: Accucore RP-MS, 2.6 µm, 50 × 2.1 mm
• Mobile Phase A: Water with 0.1% formic acid
• Mobile Phase B: Acetonitrile with 0.1% formic acid
• Gradient: 5% B (0–0.10 min) to 95% B (1.00 min), hold (1.25 min), back to 5% B (1.26 min) until 2.00 min
• Flow Rate: 1 mL/min
• Column Temperature: 40 °C
• Injection Volume: 10 µL

• LC System: Thermo Scientific Dionex UltiMate 3000 RS with HPG-3200SD pump, WPS-3000 RS autosampler, TCC-3000 RS thermostat
• MS: Thermo Scientific TSQ Vantage triple quadrupole
• Ionization: HESI-II in negative mode, spray voltage 4000 V, vaporizer 450 °C, capillary 300 °C
• Collision Pressure: 1.5 mTorr; S-lens voltages and collision energies optimized per compound

Main Results and Discussion

The method achieved separation of all five sweeteners in under one minute. MS/MS detection provided unambiguous identification without the need for full chromatographic baseline resolution. Precision assessment over six replicate injections yielded relative standard deviations below 5% for all analytes, demonstrating excellent reproducibility.

Benefits and Practical Applications

• Significantly reduced analysis time enhances laboratory throughput.
• MS/MS detection overcomes limitations of UV-inactive sweeteners.
• Low backpressure operation supports compatibility with standard HPLC systems.
• Applicable for routine QA/QC in food and beverage testing laboratories.

Future Trends and Potential Applications

• Integration with high-resolution mass spectrometry for expanded screening.
• Automation and high-throughput workflows for large-scale monitoring.
• Method adaptation to other non-chromophoric additives.
• Exploration of green solvents and miniaturized column formats.

Conclusion

The Accucore RP-MS column combined with LC-MS/MS enables rapid, precise analysis of five common artificial sweeteners in a sub-one-minute run. This approach offers a powerful tool for the fast, reliable quality control of sweetener content in diverse product matrices.