Analysis of Sugar Alcohols and Allulose Using an Arc HPLC System with Refractive Index Detection

Importance of the Topic

Overconsumption of traditional sugars contributes to obesity, diabetes and other health concerns. Sugar alcohols and allulose offer low-calorie alternatives, but accurate quantification in food, beverage and pharmaceutical matrices is vital to ensure both efficacy and safety, including avoidance of laxative effects from excessive polyol intake.

Objectives and Study Overview

This study aimed to develop and validate an isocratic high-performance liquid chromatography method with refractive index detection (HPLC-RI) for simultaneous analysis of seven common sugar alcohols (erythritol, glycerol, maltitol, mannitol, xylitol, sorbitol) and allulose in low-calorie products.

Methodology

Standards of each analyte (100 mg/mL) were diluted to calibration levels (0.16–5 mg/mL) in 50:50 acetonitrile:water. Sample preparation protocols for gums, powdered drinks, medicines and beverages involved aqueous dissolution, dilution, filtration and addition of acetonitrile. A 5 µL injection volume was eluted isocratically with 75:25 acetonitrile:water at 0.3 mL/min, with column and detector maintained at elevated temperatures to optimize retention and peak shape.

Instrumentation

• Arc HPLC system with cycle injector valve (SM FTN-R)
• Atlantis Premier BEH Z-HILIC column, 4.6×150 mm, 2.5 µm
• 2414 Refractive Index detector (10 points/sec)
• Empower 3 chromatography data software

Results and Discussion

The BEH Z-HILIC phase provided enhanced retention and distinctive selectivity for polar analytes, resolving all eight targets in a single isocratic run. Calibration curves demonstrated excellent linearity (R²>0.998). Limits of detection and quantitation met typical QA/QC requirements. Column pressure remained stable over 150 injections (<10 psi drift), and a cycle injector valve prevented sample precipitation and carryover, confirming method robustness.

Advantages and Practical Applications

The workflow delivers:

• Simultaneous quantification of multiple polyols and allulose
• Simple isocratic operation suitable for routine quality control
• High reproducibility, accuracy and precision
• No dependence on UV-active chromophores
• Reduced downtime due to injector cycle flushing

Future Trends and Opportunities

Advances may include coupling HILIC separation with mass spectrometry for trace-level screening, development of faster micro-HPLC formats for high-throughput environments, and adaptation to regulatory methods for novel sweeteners in global markets.

Conclusion

The presented Arc HPLC-RI method with Atlantis Premier BEH Z-HILIC column enables reliable, reproducible analysis of sugar alcohols and allulose in diverse matrices, supporting manufacturers and testing laboratories in standardizing reduced-sugar product assessments.

References

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4. Hossain A. et al. (2015) Rare Sugar d-allulose in Obesity and Type 2 Diabetes. Pharmacology & Therapeutics 155:49–59.
5. U.S. FDA (2020) Guidance: Declaration of Allulose and Calories. FDA.
6. Southey F. (2021) Allulose Approval in Europe. Food Navigator.