bile acids from urine - mtc-usa.com · 70000-15p-2 cogent diamond hydride™ hplc column, 100Å,...

70000-15P-2 Cogent Diamond Hydride™ HPLC Column, 100Å, 4μm, 2.1 x 150 mm Cat. No. Description APP-A55 APPLICATION NOTES MTC • MicroSolv Technology Corporation 1 Industrial Way West, Bldg E, Eatontown, NJ 07724 USA Ph. 1-732-578-1777 • Fx. 1-732-578-9777 www.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-2 Dimensions: 2.1 x150 mm Solvents: A: DI water + 0.1% formic acid B: acetonitrile + 0.1% formic acid Mobile phase: Gradient 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 t 0 = 1.44 min 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. For more information visit www.MTC-USA.com Bile Acids from Urine 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 HPLC Figure A Figure B

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70000-15P-2 Cogent Diamond Hydride™ HPLC Column, 100Å, 4µm, 2.1 x 150 mm

Cat. No. Description

APP-A55

ICA

TIO

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.

For more information visit www.MTC-USA.com

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

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