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.-Takeda, H., M. Takahashi, T. Hara, Y. Izumi, and T. Bamba. Improved quantitation of lipid classes using supercritical fluid chromatography with a charged aerosol detector. J. Lipid Res. 2019. 60: 1465-1474.
| Original language | English |
|---|---|
| Pages (from-to) | 1465-1474 |
| Number of pages | 10 |
| Journal | Journal of Lipid Research |
| Volume | 60 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - Jan 1 2019 |
Fingerprint
All Science Journal Classification (ASJC) codes
- Biochemistry
- Endocrinology
- Cell Biology
Cite this
Improved quantitation of lipid classes using supercritical fluid chromatography with a charged aerosol detector. / Takeda, Hiroaki; Takahashi, Masatomo; Hara, Takeshi; Izumi, Yoshihiro; Bamba, Takeshi.
In: Journal of Lipid Research, Vol. 60, No. 8, 01.01.2019, p. 1465-1474.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Improved quantitation of lipid classes using supercritical fluid chromatography with a charged aerosol detector
AU - Takeda, Hiroaki
AU - Takahashi, Masatomo
AU - Hara, Takeshi
AU - Izumi, Yoshihiro
AU - Bamba, Takeshi
PY - 2019/1/1
Y1 - 2019/1/1
N2 - 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.-Takeda, H., M. Takahashi, T. Hara, Y. Izumi, and T. Bamba. Improved quantitation of lipid classes using supercritical fluid chromatography with a charged aerosol detector. J. Lipid Res. 2019. 60: 1465-1474.
AB - 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.-Takeda, H., M. Takahashi, T. Hara, Y. Izumi, and T. Bamba. Improved quantitation of lipid classes using supercritical fluid chromatography with a charged aerosol detector. J. Lipid Res. 2019. 60: 1465-1474.
UR - http://www.scopus.com/inward/record.url?scp=85071062345&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85071062345&partnerID=8YFLogxK
U2 - 10.1194/jlr.D094516
DO - 10.1194/jlr.D094516
M3 - Article
C2 - 31201290
AN - SCOPUS:85071062345
VL - 60
SP - 1465
EP - 1474
JO - Journal of Lipid Research
JF - Journal of Lipid Research
SN - 0022-2275
IS - 8
ER -