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HPLC Application Sheet
Glyburide and Metformin
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 |
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| 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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