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HPLC Application Sheet
Glyburide and Metformin: 2 minutes

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Separation of "Highly Polar" and "Non-Polar" Compounds in one isocratic run using Reverse Phase Solvents.

Method Conditions


Column Cogent Qx2™ Column for LC-MS
Bidentate C-18, 4 µm, 100A
Catalog No. 40018-Q20
Dimensions 2.1 x 20 mm
Mobile Phase 
A:  50:50 acetonitrile, DI water + 0.5% formic acid
B:  85:15 acetonitrile, DI water + 0.5% formic acid
Flow rate 0.3 mL/minute
Injection Volume 1 µL
Sample 100 µg/mL
Detection UV 254nm


Discussion of Chromatograms

The polar compound, Metformin, and the nonpolar compound, Glyburide, are retained on narrow bore LC-MS column. Mobile phase is optimized for LC-MS analysis. Depending on the composition of the Mobile Phase either Metformin or Glyburide can be retained longer. NO ION-PAIR HPLC METHOD IS NEEDED TO ANALYZE METFORMIN.

Note: elution order was confirmed by LC-MS, APCI+, with single ion monitoring Metformin (m/z 130) and Glyburide (m/z 369).





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