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Bile Acids from Urine
Determination of Chenodeoxycholic acid (CDCA) (bile acid) in human urine using ANP (inverse gradient)
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printable Application Sheet
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Notes: Urinary levels of bile acids are expected to be a useful index of liver function. Also bile acid concentrations in
urine obtained from patients with cerebrotendinous xanthomatosis are different from those of normal subjects and should be
useful for diagnosis and may be used for routine assays of biological fluids. In addition unique biological activities have
been identified for bile acids, especially chenodeoxycholic acid has an effect on human ovarian cancer cell lines A2780 and
A2780-CP, the last one being cisplatin resistant. CDCA may be used in treatment of ovarian cancer especially in cisplatin
resistant disease. Synthetic bile acid derivatives are lead compounds in creation of new drugs useful for prostate cancer
therapy.
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Method Conditions
| Column |
Cogent Diamond Hydride™, 4µm, 100A |
| Catalog No. |
70000-15P-2 |
| Dimensions |
2.1 x 150 mm |
| Solvents |
A: DI water + 0.1% formic acid
B: acetonitrile + 0.1% formic acid |
| Mobile Phase |
Gradient t0 = 1.44 min
| Time |
%B |
Time |
%B |
| 0.0 |
95 |
10.0 |
50 |
| 0.2 |
95 |
11.0 |
50 |
| 7.0 |
85 |
11.1 |
95 |
| 8.0 |
85 |
14.0 |
95 |
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| Flow Rate |
0.4 mL/min. |
| Sample |
Human urine - after simple extraction |
| Analyte |
Chenodeoxycholic acid 393.29994 m/z
(M+H)+, RT = 6.26 min |
| Detection |
ESI – pos- Agilent 6210 MSD TOF mass spectrometer.
Figure B: EIC – extracted ion chromatogram of selected compound and corresponding spectrum |
Discussion
The presence of an important bile acid (chenodeoxycholic acid-CDCA) in human urine was detected using a simple mobile phase, a Cogent
Diamond Hydride HPLC column and an Agilent TOF MS instrument. The column is an excellent choice for LC-MS analysis due to its very low
carbon content (~2%) background spectrum that is extremely low. In addition the special surface of the column helps to provide a fast
equilibration while using a gradient.
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