Skip to Content
MilliporeSigma
  • A new method for determination of serum cholestanol by high-performance liquid chromatography with ultraviolet detection.

A new method for determination of serum cholestanol by high-performance liquid chromatography with ultraviolet detection.

Journal of chromatography. B, Biomedical sciences and applications (2000-07-20)
G Halperin, M Elisaf, E Leitersdorf, D Harats
ABSTRACT

We developed a method for the determination of serum 5alpha-cholestan-3beta-ol (cholestanol). The sterols were derivatized to the 4'-bromobenzenesulfonyl esters and heated in isopropanol. The cholesterol-4'-bromobenzenesulfonate was solvolyzed to cholesteryl isopropyl ether, but the derivatized cholestanol did not change and could be measured in a high-performance liquid chromatographic system equipped with a UV detector at 235 nm. On the other hand, the resulting cholesteryl isopropyl ether, having different absorbance and chromatographic mobility was not detected. This method was used for measuring cholestanol levels in patients with cerebrotendinous xanthomatosis (CTX), liver cirrhosis and serum from healthy control subjects. Reproducibility, linearity and recovery tests were done on 0.3 ml of serum samples containing >2 microg/ml cholestanol, using stigmastanol as an internal standard (I.S.). Determining cholestenol by this method can be used for diagnosis and follow-up of patients with CTX and various liver diseases.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Urea-12C, 99.9 atom % 12C
Sigma-Aldrich
Urea solution, BioUltra, ~8 M in H2O
Sigma-Aldrich
Propylene glycol, ≥99.5% (GC), FCC, FG
Sigma-Aldrich
Linoleic acid, technical, 58-74% (GC)
Sigma-Aldrich
L-Aspartic acid, BioUltra, ≥99.5% (T)
Supelco
Linoleic acid, analytical standard
Sigma-Aldrich
L-Aspartic acid, BioXtra, ≥99% (HPLC)
Sigma-Aldrich
L-Aspartic acid, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
Urea solution, 40 % (w/v) in H2O
Sigma-Aldrich
Linoleic acid, liquid, BioReagent, suitable for cell culture
Sigma-Aldrich
Propylene Glycol, meets USP testing specifications
SAFC
L-Aspartic acid
Supelco
L-Aspartic acid, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Supelco
L-Aspartic acid, Pharmaceutical Secondary Standard; Certified Reference Material
Supelco
Propylene glycol, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
L-Aspartic acid, from non-animal source, meets EP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
Linoleic acid, ≥99%
Sigma-Aldrich
2,3-Butanediol, 98%
Millipore
Urea solution, suitable for microbiology, 40% in H2O
Sigma-Aldrich
1,2-Propanediol, SAJ special grade, ≥99.0%
Sigma-Aldrich
Urea, SAJ first grade, ≥98.0%
Sigma-Aldrich
1,2-Propanediol, SAJ first grade, ≥99.0%
Sigma-Aldrich
Urea, JIS special grade, ≥99.0%
Sigma-Aldrich
Urea, ≥99.0%
Urea, European Pharmacopoeia (EP) Reference Standard
Supelco
Urea, 8 M (after reconstitution with 16 mL high purity water)
Sigma-Aldrich
Creatine, anhydrous
Sigma-Aldrich
Urea, BioUltra, for molecular biology, 99% (T)
Sigma-Aldrich
Urea, powder, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
Urea, suitable for electrophoresis