A Method for the Rapid and Simultaneous Analysis of Sweeteners in Various Food Products Using the ACQUITY Arc System and ACQUITY QDa Mass Detector

Significance of the Topic

Accurate measurement of nutritive and non-nutritive sweeteners in food and beverage products is critical for consumer safety, regulatory compliance, and product consistency. Excessive sugar intake has negative health impacts, prompting the use of alternative sweeteners in various diet foods and beverages. Reliable analytical methods enable manufacturers and quality control laboratories to quantify multiple sweeteners in a single analysis, ensuring label accuracy and adherence to EU and FDA limits.

Objectives and Study Overview

• Develop a rapid UHPLC-MS method for simultaneous quantification of 12 natural and artificial non-nutritive sweeteners.
• Validate method linearity, sensitivity, and selectivity for compounds including acesulfame-K, sucralose, saccharin, aspartame, steviol glycosides, and new high-potency sweeteners.
• Demonstrate applicability in real food and beverage matrices: tabletop sweeteners, diet soda, candy, and pudding.

Methodology

• Chromatographic separation on Waters CORTECS T3 column (3×100 mm, 2.7 µm) at 40 °C using a water/acetonitrile gradient with 0.1% formic acid.
• 8.5 min runtime at 0.55 mL/min with gradient from 10% to 100% organic.
• Calibration over eight concentration levels (0.25–25 mg/L for most sweeteners; 0.5–50 mg/L for REBA and sucralose) with quadratic 1/x weighting.

Instrumentation

• ACQUITY Arc UHPLC system
• ACQUITY QDa mass detector with ESI positive/negative modes
• Waters 2998 PDA UV detector and Empower 3 software

Key Results and Discussion

• All 12 sweeteners were separated and detected within 8.5 min, with mass-based discrimination eliminating the need for full baseline resolution.
• Single ion recording in ESI+ and ESI– modes enabled sensitive detection of chromophore-lacking sweeteners such as sucralose and cyclamate.
• Calibration achieved correlation coefficients R2 > 0.996 for all analytes.
• Sucralose formed a prominent formate adduct, while REBA and stevioside exhibited dominant fragment ions.
• Analysis of commercial samples revealed expected sweetener concentrations and detected an undeclared low-level stevioside in a Reba tabletop sweetener formulation.

Benefits and Practical Applications

• Single-run UHPLC-MS method reduces analysis time and solvent consumption compared to multiple UV-based methods.
• Mass detection improves selectivity for co-eluting compounds and UV-transparent analytes.
• Easy integration into existing workflows allows routine quality control in food and beverage industries.

Future Trends and Applications

• Extension to emerging low-calorie sweeteners, sweetener metabolites, and complex matrices.
• High-throughput applications in regulatory laboratories and industrial quality assurance.
• Integration with automated sample preparation and data analysis pipelines for large-scale screening.

Conclusion

A fast, robust, and sensitive UHPLC-MS method using the ACQUITY Arc and QDa detector enables simultaneous quantification of a broad panel of sweeteners in under 10 minutes. This approach enhances analytical efficiency and reliability for quality control and regulatory compliance in the food industry.

Reference

• M. Wu et al Direct large volume injection UHPLC-MS/MS determination of sucralose and acesulfame in water J Chromatogr A 1359 (2014) 156–161.
• D. Yang and B. Chen Simultaneous determination of nonnutritive sweeteners in foods by HPLC/ESI-MS J Agric Food Chem 57 (2009) 3025.