Quantification of HDL particle concentration by calibrated ion mobility analysis

Patrick M. Hutchins, Graziella E. Ronsein, Jeffrey S. Monette, Nathalie Pamir, Jake Wimberger, Yi He, G. M. Anantharamaiah, Daniel Seung Kim, Jane E. Ranchalis, Gail P. Jarvik

Research output: Contribution to journalArticlepeer-review

65 Scopus citations


BACKGROUND: It is critical to develop new metrics to determine whether HDL is cardioprotective in humans. One promising approach is HDL particle concentration (HDL-P), the size and concentration of HDL in plasma. However, the 2 methods currently used to determine HDL-P yield concentrations that differ >5-fold. We therefore developed and validated an improved approach to quantify HDL-P, termed calibrated ion mobility analysis (calibrated IMA).

METHODS: HDL was isolated from plasma by ultracentrifugation, introduced into the gas phase with electrospray ionization, separated by size, and quantified by particle counting. We used a calibration curve constructed with purified proteins to correct for the ionization efficiency of HDL particles.

RESULTS: The concentrations of gold nanoparticles and reconstituted HDLs measured by calibrated IMA were indistinguishable from concentrations determined by orthogonal methods. In plasma of control (n=40) and cerebrovascular disease (n = 40) participants, 3 subspecies of HDL were reproducibility measured, with an estimated total HDL-P of 13.4 (2.4) μmol/L. HDL-C accounted for 48% of the variance in HDL-P. HDL-P was significantly lower in participants with cerebrovascular disease (P=0.002), and this difference remained significant after adjustment for HDL cholesterol concentrations (P = 0.02).

CONCLUSIONS: Calibrated IMA accurately determined the concentration of gold nanoparticles and synthetic HDL, strongly suggesting that the method could accurately quantify HDL particle concentration. The estimated stoichiometry of apolipoprotein A-I determined by calibrated IMA was 3-4 per HDL particle, in agreement with current structural models. Furthermore, HDL-P was associated with cardiovascular disease status in a clinical population independently of HDL cholesterol.

Original languageEnglish (US)
Pages (from-to)1393-1401
Number of pages9
JournalClinical chemistry
Issue number11
StatePublished - Nov 1 2014
Externally publishedYes

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical


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