Improved Quantitation of Lipid Classes Using Supercritical Fluid Chromatography with a Charged Aerosol Detector

Overview

Journal J Lipid Res

Publisher Elsevier

Specialty Biochemistry

Date 2019 Jun 16

PMID 31201290

Citations 7

Authors

Hiroaki Takeda

Masatomo Takahashi

Takeshi Hara

Yoshihiro Izumi

Takeshi Bamba

Affiliations

Soon will be listed here.

Abstract

Quantitatively and rapidly analyzing lipids is necessary to elucidate their biological functions. Herein, we developed a quantitative method for various lipid classes using supercritical fluid chromatography (SFC) coupled with a charged aerosol detector (CAD), providing high-throughput data analysis to detect a large number of molecules in each lipid class as one peak. Applying the CAD was useful for analyzing lipid molecules in the same lipid class with a constant response under the same mobile phase composition. First, we optimized the washing method for the diethylamine column, achieving baseline separation of lipid classes while maintaining good peak shapes. In addition, the CAD conditions (organic solvent evaporation and numerical correction of the CAD data) were optimized to improve the signal-to-noise ratio. We used an internal standard (ceramide phosphoethanolamine d17:1-12:0), which did not coelute with the lipid classes and showed high extraction efficiency. Based on a quantitative analysis of HepG2 cells, the concentration of lipid classes detected by CAD was adequate compared with that obtained by triple-quadrupole MS (QqQMS) in a previous study because the deviations of the concentrations were 0.6- to 2.3-fold. These results also supported the quantitative performance of SFC-QqQMS developed in our previous report.

Citing Articles

Quantitative Lipidomics of Biological Samples Using Supercritical Fluid Chromatography Mass Spectrometry.

Takeda H, Izumi Y, Bamba T Methods Mol Biol. 2025; 2891():131-152.

PMID: 39812980 DOI: 10.1007/978-1-0716-4334-1_7.

Validation of an HPLC-CAD Method for Determination of Lipid Content in LNP-Encapsulated COVID-19 mRNA Vaccines.

Yu X, Yu C, Wu X, Cui Y, Liu X, Jin Y Vaccines (Basel). 2023; 11(5).

PMID: 37243041 PMC: 10222414. DOI: 10.3390/vaccines11050937.

Single-Standard Quantification Strategy for Lignin Dimers by Supercritical Fluid Chromatography with Charged Aerosol Detection.

Papp D, Rukkijakan T, Lebedeva D, Nylander T, Sandahl M, Samec J Anal Chem. 2022; 95(2):1436-1445.

PMID: 36548212 PMC: 9850414. DOI: 10.1021/acs.analchem.2c04383.

Speciation Study on O-Phosphorylethanolamine and O-Phosphorylcholine: Acid-Base Behavior and Mg Interaction.

Aiello D, Cordaro M, Napoli A, Foti C, Giuffre O Front Chem. 2022; 10:864648.

PMID: 35419347 PMC: 8996081. DOI: 10.3389/fchem.2022.864648.

Achieving Absolute Molar Lipid Concentrations: A Phospholipidomics Cross-Validation Study.

Schoeny H, Rampler E, Binh Chu D, Schoeberl A, Galvez L, Blaukopf M Anal Chem. 2022; 94(3):1618-1625.

PMID: 35025205 PMC: 8792901. DOI: 10.1021/acs.analchem.1c03743.

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