Analysis of Sucralose in Beverages Using an ACQUITY™ UPLC™ H-Class PLUS with the ACQUITY UPLC Refractive Index Detector

Importance of the Topic

Sucralose is a chlorinated sucrose derivative and low-calorie sweetener widely used in foods and beverages. Due to its lack of UV chromophore, conventional UV/Vis detectors cannot detect it, necessitating alternative detection methods such as refractive index (RI), evaporative light scattering, or mass spectrometry.

Objectives and Study Overview

This study aimed to develop a simple, robust isocratic UPLC-RI method for routine quantification of sucralose in beverages using a CORTECS™ T3 column and Waters ACQUITY™ UPLC H-Class PLUS system. The method was validated across a broad concentration range and tested on commercial soft and energy drinks.

Methodology and Materials

The sucralose standard (≥98% purity) was prepared in water:methanol (80:20) with concentrations from 7.8 to 500 µg/mL. Beverage samples were diluted (1.25X to 100X), vortexed, and filtered through 0.2 µm PTFE filters. The mobile phase was isocratic 80:20 water:methanol at 1.0 mL/min, column temperature 50 °C, detector temperature 50 °C, injection volume 5–10 µL, and sample temperature 25 °C.

Instrumentation

• ACQUITY UPLC H-Class PLUS System (Quaternary Solvent Manager)
• ACQUITY UPLC Refractive Index Detector (20 points/sec sampling rate)
• CORTECS T3 Column, 2.7 µm, 3×100 mm, 120 Å
• Empower 3 Chromatography Data Software
• PTFE syringe filters (0.2 µm)

Main Results and Discussion

The method achieved baseline separation of sucralose from matrix components and provided sharp, symmetric peaks at 50 °C. The lowest calibration standard (7.8 µg/mL) exhibited high signal-to-noise and minimal peak tailing. Calibration over 7.8–500 µg/mL displayed excellent linearity (R² > 0.999). Robustness tests across 100 injections showed minimal backpressure drift (~23 psi) and consistent retention times. Analysis of beverage samples in six replicates confirmed reproducible quantitation and well-resolved sucralose peaks.

Benefits and Practical Applications

• Simple isocratic operation with no gradient re-equilibration steps
• Low detector dispersion and stable baseline for accurate results
• High precision, accuracy, and reproducibility for routine quality control
• Wide dynamic range accommodating diverse sample concentrations

Future Trends and Possibilities

Combining RI detection with mass spectrometry or advanced light scattering could improve sensitivity and specificity. Adoption of greener solvents and faster column technologies can reduce analysis time and solvent use. Real-time online monitoring during manufacturing and extending the approach to other non-chromophoric analytes are promising developments.

Conclusion

The Waters ACQUITY UPLC H-Class PLUS coupled with an RI detector and CORTECS T3 column delivers an efficient, reproducible isocratic method for sucralose analysis in beverages. Achieving excellent linearity, stability, and precision, this method is ideal for routine QC in the food and beverage industry.

Reference

Yang J., Rainville P.D. Analysis of Soft Drink Additives with No Interference from Aspartame Degradants Using Arc HPLC System with PDA Detection. Waters Application Note 720007219, 2021.