Harman, NL, Griffin, BA and Davies, IG (2013) Separation of the principal HDL subclasses using iodixanol ultracentrifugation. Journal of Lipid Research, 54. pp. 2273-2281. ISSN 1539-7262

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Abstract

HDL subclasses detection, in cardiovascular risk, has been limited due to the time-consuming nature of current techniques. We have developed a time-saving and reliable separation of the principal HDL subclasses employing iodixanol density gradient ultracentrifugation (IxDGUC) combined with digital photography. HDL subclasses were separated in 2.5 h from prestained plasma on a three-step iodixanol gradient. HDL subclass profiles were generated by digital photography and gel scan software. Plasma samples (n = 46) were used to optimize the gradient for the resolution of HDL heterogeneity and to compare profiles generated by IxDGUC with gradient gel electrophoresis (GGE); further characterization from participants (n = 548) with a range of lipid profiles was also performed. HDL subclass profiles generated by IxDGUC were comparable to those separated by GGE as indicated by a significant association between areas under the curve for both HDL2 and HDL3 (HDL2, r = 0.896, P < 0.01; HDL3, r = 0.894, P < 0.01). The method was highly reproducible, with intra- and interassay coefficient of variation percentage < 5 for percentage area under the curve HDL2 and HDL3, and < 1% for peak Rf and peak density. The method provides time-saving and cost-effective detection and preparation of the principal HDL subclasses.

Item Type:	Article
Uncontrolled Keywords:	0601 Biochemistry And Cell Biology, 1101 Medical Biochemistry And Metabolomics
Subjects:	Q Science > QD Chemistry Q Science > QH Natural history > QH301 Biology
Divisions:	Sport Studies, Leisure & Nutrition (closed 31 Aug 19)
Publisher:	American Society for Biochemsitry and Molecular Biology
Date Deposited:	11 Mar 2016 09:00
Last Modified:	03 Sep 2021 23:24
DOI or ID number:	10.1194/jlr.D037432
URI:	https://researchonline.ljmu.ac.uk/id/eprint/3152

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