Quantitative Lipoprotein Subclass and Low Molecular Weight Metabolite Analysis in Human Serum and Plasma by ¹H NMR Spectroscopy in a Multilaboratory Trial

Abstract

We report an extensive 600 MHz NMR trial of quantitative lipoprotein and small-molecule measurements in human blood serum and plasma. Five centers with eleven 600 MHz NMR spectrometers were used to analyze 98 samples including 20 quality controls (QCs), 37 commercially sourced, paired serum and plasma samples, and two National Institute of Science and Technology (NIST) reference material 1951c replicates. Samples were analyzed using rigorous protocols for sample preparation and experimental acquisition. A commercial lipoprotein subclass analysis was used to quantify 105 lipoprotein subclasses and 24 low molecular weight metabolites from the NMR spectra. For all spectrometers, the instrument specific variance in measuring internal QCs was lower than the percentage described by the National Cholesterol Education Program (NCEP) criteria for lipid testing [triglycerides <2.7%; cholesterol <2.8%; low-density lipoprotein (LDL) cholesterol <2.8%; high-density lipoprotein (HDL) cholesterol <2.3%], showing exceptional reproducibility for direct quantitation of lipoproteins in both matrixes. The average relative standard deviations (RSDs) for the 105 lipoprotein parameters in the 11 instruments were 4.6% and 3.9% for the two NIST samples, whereas they were 38% and 40% for the 37 commercially sourced plasmas and sera, respectively, showing negligible analytical compared to biological variation. The coefficient of variance (CV) obtained for the quantification of the small molecules across the 11 spectrometers was below 15% for 20 out of the 24 metabolites analyzed. This study provides further evidence of the suitability of NMR for high-throughput lipoprotein subcomponent analysis and small-molecule quantitation with the exceptional required reproducibility for clinical and other regulatory settings.

Effect of Estrogen Receptor Status on Circulatory Immune and Metabolomics Profiles of HER2-Positive Breast Cancer Patients Enrolled for Neoadjuvant Targeted Chemotherapy

“…Metabolite and lipoprotein concentrations were estimated on 1 H NOESY spectra using the Bruker IVDr platform [64]. Receiver operating characteristic (ROC) analyses, calculated using the R package "caTools", were applied to assess whether metabolite and/or cytokine concentrations could differentiate the groups of interest.…”

Nuclear Magnetic Resonance (NMR) Spectroscopy on the Effect of PCSK9 Inhibitor on Lipoprotein Particles in Patients With Acute Coronary Syndromes(ACS)

“…According to the standard operating procedure of AVANCE IVDr magnetic resonance spectrometer system( Bruker Biospin) [15,16] . The spectra were normalized to the same quantitative scale using Bruker's QuantRef manager within TopSpin which is based on the PULCON method; hence, the spectral intensity is normalized to proton concentration in units of millimoles per liter .…”

“…The spectra were normalized to the same quantitative scale using Bruker's QuantRef manager within TopSpin which is based on the PULCON method; hence, the spectral intensity is normalized to proton concentration in units of millimoles per liter . For data analysis, the study selected the commercial Bruker IVDr LIpoprotein Subclass Analysis (B.I.-LISA) method [17,18] as lipoprotein distribution prediction method, which used a PLS-2 regression model as the algorithm for spectral deconvolution [19] .This model provides information on main lipoprotein classes, including very low density lipoprotein (VLDL), intermediate density lipoprotein (IDL), LDL, and high density lipoprotein (HDL), as well as the six VLDL subclasses (VLDL-1 to VLDL-5), six LDL subclasses (LDL-1 to LDL-6), and four HDL subclasses (HDL-1 to HDL-4). Subclasses were sorted according to their increasing density and decreasing size in ascending order, respectively.…”

High-Sensitivity CRP is Independently Associated with HDL-Mediated Cholesterol Efflux Capacity and HDL Remodeling in Patients with Coronary Artery Disease

“…In the context of the result, the HDL-C measured enzymatically was designated as ENZ-HDL-C, and the HDL-C measured by NMR was described as NMR-HDL-C. The lipoprotein-distributionprediction method selected for the analysis was the commercial Bruker IVDr Lipoprotein Subclass Analysis (B.I.-LISA) method as previously described methods [23,29]. HDL-related lipoproteins were classi ed into four subclasses, labeled numerically according to decreasing size and increasing density.…”