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HPLC Application Sheet
Glyburide and Metformin: 2 minutes
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 |
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| 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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