Clinical Analysis of Steroids by HPLC

Ordering Information

HPLC Application Sheet

Using LC-MS to Measure Steroids in Clinical Studies

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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.

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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.

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