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High-Resolution Mass Spectrometry (HRMS)

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Updated: Jun 19, 2026

A Simple Fractionated Extraction Method for the Comprehensive Analysis of Metabolites, Lipids, and Proteins from a Single Sample

A Simple Fractionated Extraction Method for the Comprehensive Analysis of Metabolites, Lipids, and Proteins from a Single Sample

Published on: June 1, 2017

Comprehensive Quantitative Profiling of Less Polar Lipids in Human Plasma Using Validated Reversed-Phase

Zuzana Lásko¹, Veronika Šubrtová¹, Ondřej Peterka¹

¹Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 53210 Pardubice, Czech Republic.

Analytical Chemistry

|November 24, 2025

Summary

This summary is machine-generated.

This study introduces a new method for analyzing less polar lipids in human plasma using reversed-phase ultrahigh-performance supercritical fluid chromatography-tandem mass spectrometry (RP-UHPSFC/MS/MS). The validated technique allows for high-throughput lipid profiling and accurate quantification of various lipid species.

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Shotgun Lipidomics of Rodent Tissues

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Published on: November 18, 2022

Area of Science:

Lipidomics
Analytical Chemistry
Biochemistry

Background:

Lipid analysis is complex due to structural diversity and isomers.
High-throughput methods are needed for lipidomic studies.

Purpose of the Study:

To develop and validate a robust method for high-throughput profiling of less polar lipids in human plasma.
To enable sensitive detection and accurate quantification of multiple lipid classes and species.

Main Methods:

Reversed-phase ultrahigh-performance supercritical fluid chromatography-tandem mass spectrometry (RP-UHPSFC/MS/MS).
Folch extraction, benzoyl chloride derivatization, and liquid-liquid extraction.
Dual C18 column system for enhanced chromatographic resolution.

Main Results:

Sensitive profiling of six major lipid classes (TG, DG, MG, SE, ST, FA) in under 18 minutes.
Separation of isomeric species, including cis/trans and positional isomers.
Accurate quantification of 147 lipid species in human plasma, with validated response factors for sterol esters.

Conclusions:

The RP-UHPSFC/MS/MS method is robust, validated, and suitable for large-scale lipidomic studies.
The method offers high sensitivity and resolution for analyzing less polar lipids.
Accurate quantification necessitates the use of response factors, particularly for sterol esters.