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HPLC Application Sheet
Separation of Steroids by LC-MS
Method Conditions
| Column |
Cogent Qx2™ Column for LC-MS
UDC-Cholesterol, 4 µm, 100A |
| Catalog No. |
69069-Q20 |
| Dimensions |
2.1 x 20 mm |
| Mobile Phase |
35:65 methanol/DI water + 0.5% formic acid
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| Flow rate |
0.3 mL/minute |
| Injection Volume |
1 µL |
| Samples |
- Adrenosterone (m/z 301)
- Corticosterone (m/z 347)
- Estrone (m/z 253)
- 4-Adrostene-3, 17-dione (m/z 287)
(10 µg/mL of each dissolved in methanol)
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| Detection |
Atmospheric Pressure Chemical Ionization in positive mode: APCI+ Single Ion Monitoring |
Discussion
Separation of four steriods in the reversed phase mode using the UDC-Cholesterol Qx2™ column is easily acheived.
Retention order is determined by hydrophobic interactions as well as the shape selectivity of the cholesterol stationary
phase. Note that the mobile phase used was isocratic.
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