Analysis of Sweeteners (Sucralose and Acesulfame K)

Importance of the Topic

Artificial sweeteners such as sucralose and acesulfame K are extensively used in food and beverage formulations to reduce caloric content without compromising sweetness. Reliable analytical methods are essential for quality control, regulatory compliance and ensuring product safety. High-performance liquid chromatography (HPLC) with suitable detection provides accurate quantification of these additives in complex matrices.

Study Objectives and Overview

This application note describes the development and validation of a reversed-phase HPLC method using the Shim-pack GIST-HP C18-AQ column coupled with evaporative light scattering detection (ELSD) for the simultaneous separation and quantification of sucralose and acesulfame K. The goal was to achieve robust baseline separation within a reasonable run time while maintaining sensitivity and reproducibility.

Methodology and Instrumentation Used

• Column: Shim-pack GIST-HP C18-AQ (250 mm × 3.0 mm I.D., 3 µm particle size).
• Mobile phase: A = 10 mmol/L ammonium formate buffer; B = acetonitrile. Gradient program: start at 2% B, hold 2% B for 7 min; ramp to 35% B by 25 min; ramp to 80% B by 25.01 min; hold at 80% B until 28 min; return to 2% B at 28.01 min; re-equilibration until 38 min.
• Flow rate: 0.7 mL/min.
• Column temperature: 55 °C.
• Detection: Evaporative light scattering detector (ELSD).
• Injection volume: 20 µL.

Main Results and Discussion

The method achieved clear baseline separation of acesulfame K and sucralose within 15 minutes. Retention times were approximately 7.5 minutes for acesulfame K and 11.0 minutes for sucralose. ELSD provided stable detection for both compounds at concentrations of 400 mg/L (acesulfame K) and 60 mg/mL (sucralose). The method exhibited good repeatability with relative standard deviations below 2% for retention time and peak area. Linearity was confirmed over relevant concentration ranges, demonstrating suitability for routine analysis in quality control environments.

Benefits and Practical Applications

This HPLC-ELSD approach offers several advantages:

• Robust baseline resolution of structurally similar sweeteners.
• High reproducibility and low detection limits suitable for regulatory thresholds.
• Applicability to diverse food and beverage matrices after simple sample preparation.
• Efficient run time supporting medium-throughput laboratory workflows.

Future Trends and Applications

Emerging developments may further enhance sweetener analysis:

• Coupling UHPLC with mass spectrometry for increased sensitivity and confirmatory identification.
• Miniaturization and microfluidic HPLC to reduce solvent consumption and analysis time.
• Automated sample preparation for higher throughput and reduced manual error.
• Integration of chemometric tools for multicomponent quantification in complex matrices.

Conclusion

The described method using the Shim-pack GIST-HP C18-AQ column and ELSD detection delivers rapid, reliable quantification of sucralose and acesulfame K. It meets the demands of modern analytical laboratories for quality control and regulatory compliance in the food and beverage industry.

References

• Shimadzu Corporation. Analysis of Sweeteners (Sucralose and Acesulfame K), Application Note ERAS-1000-0372, First Edition, September 2022.