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Clinical Chemistry 54: 1087-1088, 2008; 10.1373/clinchem.2007.100107
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(Clinical Chemistry. 2008;54:1087-1088.)
© 2008 American Association for Clinical Chemistry, Inc.

Letters to the Editor

Improved HPLC Analysis of Serum 7{alpha}-Hydroxycholest-4-en-3-one, a Marker of Bile Acid Malabsorption

Martin Lenicek1,2,a, Miroslava Juklova1, Jaroslav Zelenka1, Jan Kovar3, Milan Lukas1,4, Martin Bortlik4 and Libor Vitek1,2

1 Institute of Clinical Biochemistry and Laboratory Diagnostics, 1st Faculty of Medicine, Charles University in Prague, Czech Republic
2 4th Department of Internal Medicine, 1st Faculty of Medicine, Charles University in Prague, Czech Republic
3 Center of Experimental Medicine, Institute for Clinical and Experimental Medicine, Prague, Czech Republic
4 IBD Clinical and Research Center, ISCARE I.V.F. Lighthouse, Prague, Czech Republic

aAddress correspondence to this author at: Laboratory of Hepatology, UKBLD, U Nemocnice 2, 12808 Prague 2, Czech Republic, Fax +420-22496-2532, e-mail mleni@centrum.cz

The first 20% of the full text of this article appears below.


To the Editor:

Serum concentrations of 7{alpha}-hydroxycholest-4-en-3-one (cholesten, bile acid synthesis intermediate) have been shown to correlate with the severity of bile acid malabsorption(1)(2). Current techniques for holster quantification require sophisticated instrumentation(3) or solid-phase extraction (SPE) at 64 °C(4). We developed and validated a method for measuring serum cholesten concentrations using routinely available HPLC instrumentation, focusing on the optimization of the SPE step.

Cholesten was purchased from Steraloids, and used to prepare serum calibrators containing 0–1000 µg/L. We added 30 ng of internal standard (7β-hydroxycholest-4-en-3-one, Steraloids) dissolved in 80 µL of methanol, to 1 mL of serum. Then 5 mL of chloroform:methanol (2:1, vol/vol, analytical grade, Penta) was added; the mixture was vortex-mixed vigorously, and centrifuged (2000g, 3 min, ambient temperature). The upper phase was discarded, and 2 mL of 125 mmol/L NaCl in 50% methanol (vol/vol) was added to the sample, vortex-mixed, and centrifuged as above. The lower phase was transferred to another tube . . . [Full Text of this Article]




The following articles in journals at HighWire Press have cited this article:


Home page
 J. Lipid Res.Home page
M. Lenicek, V. Komarek, M. Zimolova, J. Kovar, M. Jirsa, M. Lukas, and L. Vitek
CYP7A1 promoter polymorphism -203A>C affects bile salt synthesis rate in patients after ileal resection
J. Lipid Res., December 1, 2008; 49(12): 2664 - 2667.
[Abstract] [Full Text] [PDF]


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