|
|
HPLC Application Sheet
Using LC-MS to Measure Steroids in Clinical Studies
Note: The total ion chromatogram of a mixture of six steroids presented substantiates the resolving power of the Cogent
UDC Cholesterol column. The peak shapes are excellent (As < 1.1 for all solutes) using an isocratic mobile phase.
Click here to view
printable Application Sheet
|
Method Conditions
| Column |
Cogent UDC Cholesterol™, 4µm, 100A
Reverse Phase Mode |
| Catalog No. |
69069-7.5P |
| Dimensions |
4.6 x 75mm |
| Mobile Phase |
50:50 MeOH/DI water + 0.5% formic acid |
| Flow rate |
0.5 mL/minute |
| Injection Volume |
5 µL |
| Peaks: | | |
| Solute | Parent
ion
m/z | Other peaks |
|
| 1. Andrenosterone |
301
333 |
283 [MH + - H2O]
[MH + + O2] |
| 2. Corticosterone |
347
311
379 |
329 [MH + - H2O]
[MH + - 2H2O]
[[MH + + O2] |
| 3. 4-androstene-3,17-dione |
287 |
319 [[MH + + O2] |
| 4. 11-alpha-acetoxyprogesterone |
373 |
313 [MH + - H3CHOOH] |
| 5. Estrone |
271 |
253 [MH + - H2O] |
| 6. Estradiol |
273 |
254 [MH + - H2O] |
| Concentration: 1 mg of each in methanol + DI water |
|
|
| Detection |
APCI, Single Ion Monitoring Mass Spec |
Discussion
Serum corticosterone concentrations are of clinical significance in adrenal dysfunction. Its measurement is sometimes used
to diagnose apparent mineralcorticoid excess syndrome. It can also be used as a bio marker of malignancy in adrenal tumors.
Using the Cogent UDC Cholesterol column allows the simultaneous measurement of the two main estrogen fractions, estrone and
estradiol in breast tumor tissue. Highly sensitive assays for estrone and estradiol for measuring low levels of estrogen in
postmenopausal women, and monitoring estrogen levels in women receiving hormone replacement therapy can also be developed
using the method presented in this note.
|
