70000-15P-2 Cogent Diamond Hydride™ HPLC Column, 100Å, 4µm, 2.1 x 150 mm
Cat. No. Description
APP-A55
AP
PL
ICA
TIO
NN
OT
ES
MTC • MicroSolv Technology Corporation1 Industrial Way West, Bldg E, Eatontown, NJ 07724 USAPh. 1-732-578-1777 • Fx. 1-732-578-9777www.mtc-usa.com Email: [email protected]
Determination of Chenodeoxycholic acid (CDCA) (bile acid) in human urine using ANP (inverse gradient)
Method Conditions
Column: Cogent Diamond Hydride™ 4µm, 100Å.Catalog No.: 70000-15P-2Dimensions: 2.1 x150 mmSolvents: A: DI water + 0.1% formic acid
B: acetonitrile + 0.1% formic acidMobile phase: Gradient Time %B Time %B
0.0 95 10.0 500.2 95 11.0 507.0 85 11.1 958.0 85 14.0 95t0 = 1.44 min
Flow rate: 0.4 mL/min.Sample: Human urine – after simple extractionAnalyte: Chenodeoxycholic acid 393.29994 m/z
(M+H)+ , RT = 6.26 minDetection: ESI – pos- Agilent 6210 MSD TOF
mass spectrometer.Figure B: EIC – extracted ion chromatogram ofselected compound and corresponding spectrum
Discussion
The presence of an important bile acid (chenodeoxycholic acid-CDCA) in human urine was detected using a simple mobilephase, a Cogent Diamond Hydride HPLC column and anAgilent TOF MS instrument. The column is an excellent choicefor LC-MS analysis due to its very low carbon content (~2%)background spectrum that is extremely low. In addition thespecial surface of the column helps to provide a fastequilibration while using a gradient.
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Bile Acids from Urine
Notes:Urinary levels of bile acids are expected to be a usefulindex of liver function. Also bile acid concentrations inurine obtained from patients with cerebrotendinousxanthomatosis are different from those of normalsubjects and should be useful for diagnosis and maybe used for routine assays of biological fluids. Inaddition unique biological activities have been identifiedfor bile acids, especially chenodeoxycholic acid has aneffect on human ovarian cancer cell lines A2780 andA2780-CP, the last one being cisplatin resistant. CDCAmay be used in treatment of ovarian cancer especiallyin cisplatin resistant disease. Synthetic bile acidderivatives are lead compounds in creation of newdrugs useful for prostate cancer therapy
HPLC
Figure A
Figure B
