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Serum cholesterol measured by isotope dilution liquid chromatography tandem mass spectrometry

  • Weiyan Zhou , Hongxia Li , Jun Dong , Shu Wang , Ruiyue Yang , Chuanbao Zhang and Wenxiang Chen EMAIL logo
Published/Copyright: December 23, 2010
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Clinical Chemistry and Laboratory Medicine
From the journal Clinical Chemistry and Laboratory Medicine Volume 49 Issue 4

Abstract

Background: Accurate cholesterol measurements are essential for the prevention and management of cardiovascular diseases. Quality assurance of cholesterol measurements requires reference methods.

Methods: An isotope dilution liquid chromatography mass spectrometric (ID/LC/MS) method was developed. Serum samples were sampled volumetrically using automated dilutors, treated with potassium hydroxide and equilibrated with 3,4-13C2 cholesterol. The natural cholesterol and the internal standard were extracted with hexane and oxidized to cholest-4-en-3,6-diones with chromic acid. The oxidation products were separated on reversed phase LC and detected by tandem MS. The method was calibrated using aqueous cholesterol calibrators and the calibration function was established with a polynomial regression.

Results: The correlation coefficients of the calibration curves were always >0.9999. The coefficients of variation (CV) of the volumetric sampling and the LC/MS analysis averaged 0.22% and 0.50%, respectively, and the total measurement CV was 0.60%. Other sources of measurement uncertainty were minor. Results on certified reference materials agreed within 1% of the certified values.

Conclusions: An ID/LC/MS method for serum cholesterol has been developed. The method is simple and accurate and may be used as a candidate reference method.

Keywords: cholesterol; isotope dilution; liquid chromatography mass spectrometry
Corresponding author: Wenxiang Chen, MD, MSc, Beijing Hospital Institute of Geriatrics and National Center for Clinical Laboratories, 1 Dahua Road, Beijing 100730, P.R. China Phone: +86 10 5811 5060, Fax: +86 10 6513 2968
Received: 2010-9-4
Accepted: 2010-11-2
Published Online: 2010-12-23
Published in Print: 2011-04-01

©2011 by Walter de Gruyter Berlin New York

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https://doi.org/10.1515/CCLM.2011.093
Keywords for this article
cholesterol; isotope dilution; liquid chromatography mass spectrometry

Articles in the same Issue

  1. Editorial
  2. Nuclear and radiation accidents: the next and further challenge for laboratory diagnostics
  3. Reviews
  4. Diagnostic, prognostic and therapeutic relevance of B-type natriuretic hormone and related peptides in children with congenital heart diseases
  5. Miniaturization and globalization of clinical laboratory activities
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  14. Genetics and Molecular Diagnostics
  15. Non-invasive prenatal diagnosis of trisomy 21 by reverse transcriptase multiplex ligation-dependent probe amplification
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