Application of directly coupled HPLC MMR to separation and characterization of lipoproteins from human serum

Research output: Contribution to journalJournal articleResearchpeer-review

Standard

Application of directly coupled HPLC MMR to separation and characterization of lipoproteins from human serum. / Daykin, C. A.; Corcoran, O.; Hansen, S. H.; Bjornsdottir, I.; Cornett, Claus; Connor, S. C.; Lindon, J. C.; Nicholson, J. K.

In: Analytical Chemistry, Vol. 73, No. 6, 2001, p. 1084-1090.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Daykin, CA, Corcoran, O, Hansen, SH, Bjornsdottir, I, Cornett, C, Connor, SC, Lindon, JC & Nicholson, JK 2001, 'Application of directly coupled HPLC MMR to separation and characterization of lipoproteins from human serum', Analytical Chemistry, vol. 73, no. 6, pp. 1084-1090.

APA

Daykin, C. A., Corcoran, O., Hansen, S. H., Bjornsdottir, I., Cornett, C., Connor, S. C., ... Nicholson, J. K. (2001). Application of directly coupled HPLC MMR to separation and characterization of lipoproteins from human serum. Analytical Chemistry, 73(6), 1084-1090.

Vancouver

Daykin CA, Corcoran O, Hansen SH, Bjornsdottir I, Cornett C, Connor SC et al. Application of directly coupled HPLC MMR to separation and characterization of lipoproteins from human serum. Analytical Chemistry. 2001;73(6):1084-1090.

Author

Daykin, C. A. ; Corcoran, O. ; Hansen, S. H. ; Bjornsdottir, I. ; Cornett, Claus ; Connor, S. C. ; Lindon, J. C. ; Nicholson, J. K. / Application of directly coupled HPLC MMR to separation and characterization of lipoproteins from human serum. In: Analytical Chemistry. 2001 ; Vol. 73, No. 6. pp. 1084-1090.

Bibtex

@article{f85b701c17624371ab7821220bc16812,
title = "Application of directly coupled HPLC MMR to separation and characterization of lipoproteins from human serum",
abstract = "Disorders in lipoprotein metabolism are critical in the etiology of several disease states such as coronary heart disease and atherosclerosis, Thus, there is considerable interest in the development of novel methods for the analysis of lipoprotein complexes. We report here a simple chromatographic method for the separation of highdensity lipoprotein, low-density lipoprotein, and very low-density lipoprotein from intact serum or plasma. The separation was achieved using a hydroxyapatite column and elution with pH 7.4 phosphate buffer with 100-muL injections of whole plasma. Coelution of HDL with plasma proteins such as albumin occurred, and this clearly limits quantitation of that species by HPLC peak integration. We also show, for the first time, the application of directly coupled HPLC H-1 NMR spectroscopy to confirm the identification of the three major lipoproteins, The full chromatographic run time was 90 min with stopped-now 600-MHz NMR spectra of each lipoprotein being collected using 128 scans, in 7 min. The H-1 NMR chemical shifts of lipid signals were identical to conventional NMR spectra of freshly prepared lipoprotein standards, confirming that the lipoproteins were not degraded by the HPLC separation and that their gross supramolecular organization was intact.",
keywords = "performance liquid-chromatography nuclear-magnetic-resonance human blood-plasma hydroxyapatite chromatography proteins quantitation spectroscopy variability density",
author = "Daykin, {C. A.} and O. Corcoran and Hansen, {S. H.} and I. Bjornsdottir and Claus Cornett and Connor, {S. C.} and Lindon, {J. C.} and Nicholson, {J. K.}",
year = "2001",
language = "Udefineret/Ukendt",
volume = "73",
pages = "1084--1090",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "6",

}

RIS

TY - JOUR

T1 - Application of directly coupled HPLC MMR to separation and characterization of lipoproteins from human serum

AU - Daykin, C. A.

AU - Corcoran, O.

AU - Hansen, S. H.

AU - Bjornsdottir, I.

AU - Cornett, Claus

AU - Connor, S. C.

AU - Lindon, J. C.

AU - Nicholson, J. K.

PY - 2001

Y1 - 2001

N2 - Disorders in lipoprotein metabolism are critical in the etiology of several disease states such as coronary heart disease and atherosclerosis, Thus, there is considerable interest in the development of novel methods for the analysis of lipoprotein complexes. We report here a simple chromatographic method for the separation of highdensity lipoprotein, low-density lipoprotein, and very low-density lipoprotein from intact serum or plasma. The separation was achieved using a hydroxyapatite column and elution with pH 7.4 phosphate buffer with 100-muL injections of whole plasma. Coelution of HDL with plasma proteins such as albumin occurred, and this clearly limits quantitation of that species by HPLC peak integration. We also show, for the first time, the application of directly coupled HPLC H-1 NMR spectroscopy to confirm the identification of the three major lipoproteins, The full chromatographic run time was 90 min with stopped-now 600-MHz NMR spectra of each lipoprotein being collected using 128 scans, in 7 min. The H-1 NMR chemical shifts of lipid signals were identical to conventional NMR spectra of freshly prepared lipoprotein standards, confirming that the lipoproteins were not degraded by the HPLC separation and that their gross supramolecular organization was intact.

AB - Disorders in lipoprotein metabolism are critical in the etiology of several disease states such as coronary heart disease and atherosclerosis, Thus, there is considerable interest in the development of novel methods for the analysis of lipoprotein complexes. We report here a simple chromatographic method for the separation of highdensity lipoprotein, low-density lipoprotein, and very low-density lipoprotein from intact serum or plasma. The separation was achieved using a hydroxyapatite column and elution with pH 7.4 phosphate buffer with 100-muL injections of whole plasma. Coelution of HDL with plasma proteins such as albumin occurred, and this clearly limits quantitation of that species by HPLC peak integration. We also show, for the first time, the application of directly coupled HPLC H-1 NMR spectroscopy to confirm the identification of the three major lipoproteins, The full chromatographic run time was 90 min with stopped-now 600-MHz NMR spectra of each lipoprotein being collected using 128 scans, in 7 min. The H-1 NMR chemical shifts of lipid signals were identical to conventional NMR spectra of freshly prepared lipoprotein standards, confirming that the lipoproteins were not degraded by the HPLC separation and that their gross supramolecular organization was intact.

KW - performance liquid-chromatography nuclear-magnetic-resonance human blood-plasma hydroxyapatite chromatography proteins quantitation spectroscopy variability density

M3 - Tidsskriftartikel

VL - 73

SP - 1084

EP - 1090

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 6

ER -

ID: 38061262