Concurrent profiling of polar metabolites and lipids in human plasma using HILIC-FTMS

Abstract

Blood plasma is the most popularly used sample matrix for metabolite profiling studies, which aim to achieve global metabolite profiling and biomarker discovery. However, most of the current studies on plasma metabolite profiling focused on either the polar metabolites or lipids. In this study, a comprehensive analysis approach based on HILIC-FTMS was developed to concurrently examine polar metabolites and lipids. The HILIC-FTMS method was developed using mixed standards of polar metabolites and lipids, the separation efficiency of which is better in HILIC mode than in C5 and C18 reversed phase (RP) chromatography. This method exhibits good reproducibility in retention times (CVs < 3.43%) and high mass accuracy (<3.5 ppm). In addition, we found MeOH/ACN/Acetone (1:1:1, v/v/v) as extraction cocktail could achieve desirable gathering of demanded extracts from plasma samples. We further integrated the MeOH/ACN/Acetone extraction with the HILIC-FTMS method for metabolite profiling and smoking-related biomarker discovery in human plasma samples. Heavy smokers could be successfully distinguished from non smokers by univariate and multivariate statistical analysis of the profiling data, and 62 biomarkers for cigarette smoke were found. These results indicate that our concurrent analysis approach could be potentially used for clinical biomarker discovery, metabolite-based diagnosis, etc.

Pediatric Multi-Organ Dysfunction Syndrome: Analysis by an Untargeted “Shotgun” Lipidomic Approach Reveals Low-abundance Plasma Phospholipids and Dynamic Recovery Over 8-Day Period, a Single-Center Observational Study

“…Blood samples collected in EDTA-treated tubes were immediately placed on ice and then spun at 4oC (once for 15 minutes at 1500rpm; a second spin for 10 minutes at 10,000rpm), plasma was harvested and frozen to −20oC, and −80oC for long-term. Lipidome profiles were determined from thawed plasma, subjected to lipid extraction with acetone, methanol, and acetonitrile [29], followed by analysis by nano-electrospray direct infusion high resolution/accurate mass spectrometry and tandem mass spectrometry utilizing an LTQ-Orbitrap Velos mass spectrometer (Thermo Scientific, Waltham, MA) with the FT analyzer operating at 100,000 resolving power over two minutes. An Advion Nanomate Triversa (Advion Biosciences, Ithaca, NY) served as the nano-electrospray source with high-throughput autosampler.…”

Phosphate transporter PstSCAB of Campylobacter jejuni is essential for lactate-dependent growth and colonization in chickens

“…Ten microliters of each metabolite extract was injected and separated on a Phenomenex HILIC LC column, 2.0 mm x 100 mm, 3.0 micron particle size, 100 Angstrom pore size, using a gradient of (A) water containing 50 mM ammonium formate, and (B) acetonitrile from 95% to 50% B over 14 minutes as previously descrived. (38). A guard cartridge of matching chemistry was fitted to the analytical column.…”

Lipidomics unveils the complexity of the lipidome in metabolic diseases

“…Monophasic extraction strategies may also simultaneously facilitate recovery of polar metabolites to enable analysis of the entire metabolome. Monophasic extraction methods include variants of the methanol/chloroform/water mixtures discussed above or methanol, acetonitrile, and acetone [ 11 , 12 ]. After extraction, the lipids are often separated by various chromatographic methods such as thin layer chromatography (TLC), gas chromatography (GC), and high-performance liquid chromatography (HPLC), or lipids can be directly infused to a mass spectrometer for detection without separation.…”