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HPLC Application Sheet
Glyburide and Metformin

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

Method Conditions


Column Cogent Bidentate C-18, 4 µm, 100A
Catalog No. 40018-75P
Dimensions 4.6 x 75 mm
Mobile Phase 
A:  85:15 acetonitrile,
DI water + 0.5% formic acid
B:  50:50 acetonitrile,
DI water + 0.5% formic acid
Flow rate 0.5 mL/minute
Injection Volume 1 µL
Sample 100 µg/µL
Detection UV 254nm


Discussion of Chromatograms

The polar compound, Metformin, and the nonpolar compound, Glyburide, can be retained on the same stationary phase (see A & B), utilizing the unique properties of the hydride based Cogent Bidentate C18 column. Depending on the mobile phase composition either Metformin or Glyburide can be retained longer.

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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