Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter November 14, 2013

A simple and precise method for direct measurement of fractional esterification rate of high density lipoprotein cholesterol by high performance liquid chromatography

  • Jun Dong , Songlin Yu , Ruiyue Yang , Hongxia Li , Hanbang Guo , Haijian Zhao , Shu Wang and Wenxiang Chen EMAIL logo

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.


Corresponding author: Prof. Wenxiang Chen, Beijing Hospital Institute of Geriatrics and National Center for Clinical Laboratories, Beijing 100730, P.R. China, Phone: +86 105811 5060, Fax: +86 10 6513 2968, E-mail:

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.

References

1. Glomset JA. The mechanism of the plasma cholesterol esterification reaction: plasma fatty acid transferase. Biochim Biophys Acta 1962;65:128–35.10.1016/0006-3002(62)90156-7Search in Google Scholar

2. Glomset JA, Janssen ET, Kennedy R, Dobbins J. Role of plasma lecithin:cholesterol acyltransferase in the metabolism of high density lipoproteins. J Lipid Res 1966;7:638–48.10.1016/S0022-2275(20)39245-2Search in Google Scholar

3. Calabresi L, Franceschini G. Lecithin:cholesterol acyltransferase, high-density lipoproteins, and atheroprotection in humans. Trends Cardiovasc Med 2010;20:50–3.10.1016/j.tcm.2010.03.007Search in Google Scholar

4. Kunnen S, Van Eck M. Lecithin:cholesterol acyltransferase: old friend or foe in atherosclerosis? J Lipid Res 2012;53:1783–99.10.1194/jlr.R024513Search in Google Scholar

5. Dobiasova M, Frohlich JJ. Structural and functional assessment of high density lipoprotein heterogeneity. Clin Chem 1994;40:1554–8.10.1093/clinchem/40.8.1554Search in Google Scholar

6. Dobiasova M, Stribrna J, Pritchard PH, Frohlich JJ. Cholesterol esterification rate in plasma depleted of very low and low density lipoproteins is controlled by the proportion of HDL2 and HDL3 subclasses: study in hypertensive and normal middle-aged and septuagenarian men. J Lipid Res 1992;33:1411–8.10.1016/S0022-2275(20)41395-1Search in Google Scholar

7. Ohta T, Saku K, Takata K, Nagata N, Maung KK, Matsuda I. Fractional esterification rate of cholesterol in high density lipoprotein (HDL) can predict the particle size of low density lipoprotein and HDL in patients with coronary heart disease. Atherosclerosis 1997;135:205–12.10.1016/S0021-9150(97)00163-9Search in Google Scholar

8. Ohta T, Kakiuti Y, Kurahara K, Saku K, Nagata N, Matsuda I. Fractional esterification rate of cholesterol in high density lipoprotein is correlated with low density lipoprotein particle size in children. J Lipid Res 1997;38:139–46.10.1016/S0022-2275(20)37283-7Search in Google Scholar

9. Frohlich J, Dobiasova M. Fractional esterification rate of cholesterol and ratio of triglycerides to HDL cholesterol are powerful predictors of positive findings on coronary angiography. Clin Chem 2003;49:1873–80.10.1373/clinchem.2003.022558Search in Google Scholar PubMed

10. Dobiasova M, Frohlich J, Sedova M, Cheung MC, Brown BG. Cholesterol esterification and atherogenic index of plasma correlate with lipoprotein size and findings on coronary angiography. J Lipid Res 2011;52:566–71.10.1194/jlr.P011668Search in Google Scholar PubMed PubMed Central

11. Brown BG, Zhao XQ, Chait A, Fisher LD, Cheung MC, Morse JS, et al. Simvastatin and niacin, antioxidant vitamins, or the combination for the prevention of coronary disease. N Engl J Med 2001;345:1583–92.10.1056/NEJMoa011090Search in Google Scholar

12. Saku K, Zhang B, Ohta T, Arakawa K. Quantity and function of high density lipoprotein as an indicator of coronary atherosclerosis. J Am Coll Cardiol 1999;33:436–43.10.1016/S0735-1097(98)00560-9Search in Google Scholar

13. Lichtenstein AH, Ausman LM, Jalbert SM, Vilella-Bach M, Jauhiainen M, McGladdery S, et al. Efficacy of a therapeutic lifestyle change/step 2 diet in moderately hypercholesterolemic middle-aged and elderly female and male subjects. J Lipid Res 2002;43:264–73.10.1016/S0022-2275(20)30168-1Search in Google Scholar

14. Dobiasova M, Schutzova M. Cold labelled substrate and estimation of cholesterol esterification rate in lecithin cholesterol acyltransferase radioassay. Physiol Bohemoslov 1986;35:319–27.Search in Google Scholar

15. Dobiasova M, Adler L, Ohta T, Frohlich J. Effect of labeling of plasma lipoproteins with [(3)H]cholesterol on values of esterification rate of cholesterol in apolipoprotein B-depleted plasma. J Lipid Res 2000;41:1356–7.10.1016/S0022-2275(20)33444-1Search in Google Scholar

16. Dong J, Chen WX, Wang S, Zhang J, Li H, Guo H, et al. Jones oxidation and high performance liquid chromatographic analysis of cholesterol in biological samples. J Chromatogr B Analyt Technol Biomed Life Sci 2007;858:239–46.10.1016/j.jchromb.2007.08.038Search in Google Scholar

17. Warnick GR, Benderson J, Albers JJ. Dextran sulfate-Mg2+ precipitation procedure for quantitation of high-density-lipoprotein cholesterol. Clin Chem 1982;28:1379–88.10.1093/clinchem/28.6.1379Search in Google Scholar

18. Lopes-Virella MF, Stone P, Ellis S, Colwell JA. Cholesterol determination in high-density lipoproteins separated by three different methods. Clin Chem 1977;23:882–4.10.1093/clinchem/23.5.882Search in Google Scholar

19. Dong J, Guo HB, Yang RY, Li H, Wang S, Zhang J, et al. Serum LDL- and HDL-cholesterol determined by ultracentrifugation and HPLC. J Lipid Res 2011;52:383–8.10.1194/jlr.D008979Search in Google Scholar

20. Dong J, Guo H, Yang R, Li H, Wang S, Zhang J, et al. A novel and precise method for simultaneous measurement of serum HDL and LDL subfractions and lipoprotein (a) cholesterol by ultracentrifugation and high-performance liquid chromatography. Clin Chim Acta 2012;413:1071–6.10.1016/j.cca.2012.02.022Search in Google Scholar

21. Fraser CG. Biological variation: from principles to practice. Washington, DC: AACC Press, 2001.Search in Google Scholar

22. Glomset JA, Wright JL. Some properties of a cholesterol esterifying enzyme in human plasma. Biochim Biophys Acta 1964;89:266–76.Search in Google Scholar

23. Kobori K, Saito K, Ito S, Kotani K, Manabe M, Kanno T. A new enzyme-linked immunosorbent assay with two monoclonal antibodies to specific epitopes measures human lecithin-cholesterol acyltransferase. J Lipid Res 2002;43:325–34.10.1016/S0022-2275(20)30175-9Search in Google Scholar

24. Stokke KT, Norum KR. Determination of lecithin: cholesterol acyltransfer in human blood plasma. Scand J Clin Lab Invest 1971;27:21–7.10.3109/00365517109080184Search in Google Scholar

25. Albers JJ, Chen CH, Adolphson JL. Lecithin:cholesterol acyltransferase (LCAT) mass; its relationship to LCAT activity and cholesterol esterification rate. J Lipid Res 1981;22:1206–13.10.1016/S0022-2275(20)37313-2Search in Google Scholar

26. Pritchard PH. Lecithin: cholesterol acyltransferase. In: Rifai N, Warnick GR, Dominiczak MH, editor. Handbook of lipoprotein testing. Washington DC: AACC Press, 1997:401–14.Search in Google Scholar

27. Dobiasova M, Frohlich J. Measurement of fractional esterification rate of cholesterol in plasma depleted of apolipoprotein B containing lipoprotein: methods and normal values. Physiol Res 1996;45:65–73.Search in Google Scholar

28. Dobiasova M, Schutzova M. Factors influencing the cholesterol esterification rate in lecithin cholesterol acyltransferase radioassay. Physiol Bohemoslov 1986;35:464–72.Search in Google Scholar

29. Dobiasova M, Stribrna J, Sparks DL, Pritchard PH, Frohlich JJ. Cholesterol esterification rates in very low density lipoprotein- and low density lipoprotein-depleted plasma. Relation to high density lipoprotein subspecies, sex, hyperlipidemia, and coronary artery disease. Arterioscler Thromb 1991;11:64–70.10.1161/01.ATV.11.1.64Search in Google Scholar

30. Dobiasova M, Raslova K, Rauchova H, Vohnout B, Ptackova K, Frohlich J. Atherogenic lipoprotein profile in families with and without history of early myocardial infarction. Physiol Res 2001;50:1–8.Search in Google Scholar

Received: 2013-7-10
Accepted: 2013-10-14
Published Online: 2013-11-14
Published in Print: 2014-04-01

©2014 by Walter de Gruyter Berlin/Boston

Downloaded on 28.3.2024 from https://www.degruyter.com/document/doi/10.1515/cclm-2013-0525/html
Scroll to top button