Next Article in Journal
Recent High Performance Thin Layer Chromatographic Studies on Biomphalaria glabrata (Gastropoda)
Previous Article in Journal
Recent Advances and Uses of Monolithic Columns for the Analysis of Residues and Contaminants in Food
Previous Article in Special Issue
Analyses of Phytohormones in Coconut (Cocos Nucifera L.) Water Using Capillary Electrophoresis-Tandem Mass Spectrometry
Please note that, as of 1 January 2016, Chromatography has been renamed to Separations and is now published here.
Open AccessArticle

Multivariate DoE Optimization of Asymmetric Flow Field Flow Fractionation Coupled to Quantitative LC-MS/MS for Analysis of Lipoprotein Subclasses

Division of Laboratory Sciences, Centers for Disease Control and Prevention, National Center for Environmental Health, Atlanta, GA 30341, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Gianni Galaverna
Chromatography 2015, 2(1), 96-117; https://doi.org/10.3390/chromatography2010096
Received: 12 December 2014 / Revised: 21 January 2015 / Accepted: 11 February 2015 / Published: 26 February 2015
(This article belongs to the Special Issue Advances in Hyphenated Methods in Separation)
In this report we demonstrate a practical multivariate design of experiment (DoE) approach for asymmetric flow field-flow fractionation (AF4) method optimization using separation of lipoprotein subclasses as an example. First, with the aid of commercially available software, we built a full factorial screening design where the theoretical outcomes were calculated by applying established formulas that govern AF4 channel performance for a 5–35 nm particle size range of interest for lipid particles. Second, using the desirable ranges of instrumental parameters established from theoretical optimization, we performed fractional factorial DoE for AF4 separation of pure albumin and ferritin with UV detection to narrow the range of instrumental parameters and allow optimum size resolution while minimizing losses from membrane immobilization. Third, the optimal range of conditions were tested using response surface DoE for sub-fractionation of high and low density lipoproteins (HDL and LDL) in human serum, where the recovery of the analytes were monitored by fraction collection and isotope-dilution LC-MS/MS analysis of each individual fraction for cholesterol and apolipoproteins (ApoA-1 and ApoB-100). Our results show that DoE is an effective tool in combining AF4 theoretical knowledge and experimental data in finding the most optimal set of AF4 instrumental parameters for quantitative coupling with LC-MS/MS measurements. View Full-Text
Keywords: asymmetric flow field flow fractionation; mass spectrometry; lipoproteins; design of experiment; quality by design asymmetric flow field flow fractionation; mass spectrometry; lipoproteins; design of experiment; quality by design
Show Figures

Figure 1

MDPI and ACS Style

Kuklenyik, Z.; Gardner, M.S.; Parks, B.A.; Schieltz, D.M.; Rees, J.C.; McWilliams, L.G.; Williamson, Y.M.; Pirkle, J.L.; Barr, J.R. Multivariate DoE Optimization of Asymmetric Flow Field Flow Fractionation Coupled to Quantitative LC-MS/MS for Analysis of Lipoprotein Subclasses. Chromatography 2015, 2, 96-117.

Show more citation formats Show less citations formats

Article Access Map by Country/Region

1
Only visits after 24 November 2015 are recorded.
Back to TopTop