Separation of Metformin and Voglibose Using a Thermo Scientific Accucore HILIC HPLC Column

Importance of the Topic

Polar anti-diabetic agents such as metformin and voglibose lack strong UV chromophores, making conventional HPLC separation and detection challenging. Efficient and rapid separation of these compounds supports therapeutic monitoring and quality control in pharmaceutical development.

Objectives and Study Overview

This study aimed to develop a rapid, reproducible HILIC-based method to separate metformin and voglibose using a Thermo Scientific Accucore HILIC column coupled with tandem mass spectrometric detection.

Methodology

A working standard containing 1 µg/mL metformin and 2 µg/mL voglibose was prepared in acetonitrile/water/formic acid. Chromatography was performed under the following conditions:

• Column: Accucore HILIC, 2.6 µm, 50 × 2.1 mm
• Mobile phase: 17:83 (v/v) 100 mM ammonium formate (pH 3.2)/acetonitrile
• Flow rate: 0.4 mL/min, ambient temperature, 5 µL injection
• Run time: 3.5 min

Used Instrumentation

• Thermo Scientific Accela 1250 pump with Open Autosampler
• Thermo Scientific TSQ Vantage triple quadrupole mass spectrometer (HESI source, positive mode)
• Thermo Scientific LCQuan software for data processing

Main Results and Discussion

Metformin eluted at approximately 1.05 min (k′≈2.9) and voglibose at 1.96 min (k′≈6.3) with retention time RSDs below 0.6% over six injections. High peak efficiency and minimal tailing were achieved due to the core-shell particle design reducing secondary interactions. Total analysis time fell below four minutes, demonstrating suitability for high-throughput workflows.

Benefits and Practical Applications

• Rapid, reproducible separation of polar analytes
• Enhanced sensitivity with MS detection compensates for lack of chromophores
• Core-shell HILIC column provides high efficiency and robustness
• Applicable to QC, pharmacokinetic studies, and routine monitoring

Future Trends and Potential Applications

Advances may include coupling HILIC with high-resolution MS for untargeted metabolomics, method miniaturization for microflow applications, and expansion to other polar drug combinations in clinical and regulatory settings.

Conclusion

The developed Accucore HILIC–MS method offers a fast and reliable approach for simultaneous separation of metformin and voglibose, supporting efficient pharmaceutical analysis in research and quality control environments.

References

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