A simple and precise method for direct measurement of fractional esterification rate of high density lipoprotein cholesterol by high performance liquid chromatography
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Jun Dong
Abstract
Background: The relationship between fractional cholesterol esterification rate in plasma or serum high-density lipoprotein (HDL) (FERHDL) and lipoprotein subfractions and other cardiovascular disease (CVD) risk factors has been demonstrated. However, the current method for measuring FERHDL requires fresh serum samples and radioactive labeling of the samples, making it impractical for use in clinical laboratories. In this study, we developed a simple and precise HPLC method for the measurement of FERHDL. Correlations between FERHDL and CVD risk factors were evaluated in 119 healthy volunteers.
Methods: Fasting blood samples were collected and serum was isolated within 2 h. Serum HDL was prepared by precipitation of apolipoprotein B (apoB)-containing lipoproteins with dextran sulfate and magnesium chloride. HDL fractions were divided into two aliquots and incubated at 0°C and 37°C, respectively, for 1 h. Free cholesterol in the HDL fractions was analyzed by HPLC. FERHDL was calculated as the percent decrease of free cholesterol during incubation.
Results: The esterification reaction of HDL free cholesterol was not linear, but the measured FERHDL was stable when serum samples were stored at room temperature for <4 h, or at 4°C for <24 h. The intra-assay and total CVs for FERHDL measurements were 1.0%–2.1% and 1.6%–3.8%, respectively. Results of 119 healthy volunteers showed that FERHDL was positively correlated with age, BMI, blood pressure, total cholesterol (TC), triglyceride (TG) and small dense low-density lipoprotein-cholesterol (LDLb-C), and negatively correlated with HDL-C and HDL2-C. FERHDL has shown to be a predictor of HDL and LDL subfraction distributions.
Conclusions: This method is simple, non-radioactive and precise and will be useful in prediction of lipoprotein subfraction distributions and in clinical assessment of CVD risks.
Acknowledgments
This study was supported by research grants from National Science & Technology Pillar Program under the Twelfth Five-year Plan of China (2012BAI37B01), and National Natural Science Foundation of China (30872413, 81171647).
Conflict of interest statement
Authors’ conflict of interest disclosure: The authors stated that there are no conflicts of interest regarding the publication of this article. Research funding played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
Research funding: None declared.
Employment or leadership: None declared.
Honorarium: None declared.
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Artikel in diesem Heft
- Masthead
- Masthead
- Editorials
- Inflammatory bowel diseases: where we are and where we should go
- Thrombophilia testing. Useful or hype?
- Reviews
- Inflammatory bowel diseases: from pathogenesis to laboratory testing
- Crohn’s disease specific pancreatic antibodies: clinical and pathophysiological challenges
- Point/Counterpoint
- The utility of thrombophilia testing
- The futility of thrombophilia testing
- Genetics and Molecular Diagnostics
- Identification of an 18 bp deletion in the TWIST1 gene by CO-amplification at lower denaturation temperature-PCR (COLD-PCR) for non-invasive prenatal diagnosis of craniosynostosis: first case report
- General Clinical Chemistry and Laboratory Medicine
- Agreement of seven 25-hydroxy vitamin D3 immunoassays and three high performance liquid chromatography methods with liquid chromatography tandem mass spectrometry
- First trimester pregnancy-associated plasma protein A and human chorionic gonadotropin-beta in early and late pre-eclampsia
- S100B blood level measurement to exclude cerebral lesions after minor head injury: the multicenter STIC-S100 French study
- NGAL, L-FABP, and KIM-1 in comparison to established markers of renal dysfunction
- Value-added reporting of antinuclear antibody testing by automated indirect immunofluorescence analysis
- Cardiovascular Diseases
- A fit-for-purpose approach to analytical sensitivity applied to a cardiac troponin assay: time to escape the ‘highly-sensitive’ trap
- Diabetes
- A simple and precise method for direct measurement of fractional esterification rate of high density lipoprotein cholesterol by high performance liquid chromatography
- Infectious Diseases
- Detection of unamplified HCV RNA in serum using a novel two metallic nanoparticle platform
- Increased plasma arginase activity in human sepsis: association with increased circulating neutrophils
- Corrigendum
- The relationship between estimated average glucose and fasting plasma glucose
- Letter to the Editors
- The obsessive-compulsory clinical pathologist
- Prism effect of specimen receiving – generation of fundamentals for the smooth progress of analytical processing
- Preanalytical errors: the professionals’ perspective
- Corrected reports in laboratory medicine in a Chinese university hospital for 3 years
- Comparison of an enzymatic assay with liquid chromatography-pulsed amperometric detection for the determination of lactulose and mannitol in urine of healthy subjects and patients with active celiac disease
- Effect of freezing-thawing process on neuron specific enolase concentration in severe traumatic brain injury sera samples
- Undetected creatinine levels
- The effect of centrifugation on three urine protein assays: benzethonium chloride, benzalkonium chloride and pyrogallol red
