Simultaneous Determination of 15 Saccharides Without Interference by Salt in Seasonings

Importance of the Topic

The growing awareness of the impact of dietary sugars on conditions such as diabetes, obesity and dementia has driven the need for reliable sugar analysis in foods. Seasonings often contain hidden sugars alongside high salt levels, which can interfere with conventional detectors. This study develops an approach to accurately quantify 15 saccharides in complex, salted matrices.

Objectives and Study Overview

The aim was to establish and validate a gradient hydrophilic interaction liquid chromatography (HILIC) method combined with an ELSD-LT III detector on a Shimadzu Nexera™ lite system. Fifteen saccharides from monosaccharides to tetrasaccharides were targeted. Applications included six types of commercially available seasonings, with desalination steps to remove salt interference.

Methodology and Instrumentation

• Chromatography: Shodex HILICpak VG-50 4E column (250 × 4.6 mm, 5 μm).
• Instrument: Shimadzu Nexera lite HPLC with ELSD-LT III (filter 4 s, drift tube 40 °C, N₂ at 350 kPa).
• Mobile phase: water (A) and acetonitrile (B) gradient from 88% B to 60% B over 40 min at 1.0 mL/min, column at 45 °C.
• Sample prep: extraction with 70% acetonitrile, centrifugation, filtration; desalination via H⁺-type SPE (Maxi-Clean IC-H) for high-salt samples.

Main Results and Discussion

• All 15 saccharides eluted within ~40 min; glucose and galactose were baseline separated despite isomeric similarity.
• Repeatability: retention time RSD ≤ 0.2%, peak area RSD ≤ 6.3% (n=6).
• Linearity: log–log ELSD calibration curves showed r² ≥ 0.998 over concentration ranges from 10 to 1000 mg/L.
• Recovery in spiked ketchup samples: 82–94% for fructose, glucose and sucrose.
• Desalination removed salt peaks, enabling quantitation of low-level sugars (maltose, isomaltose) in steak sauce, ketchup, soy sauce and vinegar.
• Quantified sugar levels varied widely: e.g., BBQ sauce contained ~542 mg/L fructose and ~597 mg/L glucose; balsamic vinegar showed high sugar content without significant salt interference.

Benefits and Practical Applications

• Sensitive, simultaneous analysis of mono- to tetrasaccharides in salted foods.
• Effective separation of critical isomers, ensuring accurate quantitation.
• Integrated desalination step overcomes salt interference, broadening application to various food matrices.
• Suitable for food industry QA/QC, nutritional labeling, and research on sugar–health relationships.

Future Trends and Potential Uses

The HILIC-ELSD protocol can be adapted for other sugar classes and salted matrices. Integration with mass spectrometry could enhance structural identification. Automation of desalination procedures and extension to glycan profiling in clinical and nutritional studies represent promising directions.

Conclusion

This work demonstrates a robust HILIC-ELSD method on the Nexera lite platform for the simultaneous determination of 15 saccharides in high-salt seasonings. The use of cation-exchange SPE effectively removes salt interference, delivering precise, reproducible sugar quantitation and confirming the method’s broad applicability in food analysis.

Reference

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