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
N1 - Funding Information:
This work was supported by Japan Agency for Medical Research and Development Grants 19gm0910010h0204, 19gm0910013h0003, and 19gm1010010s0202 (Y.I., T.B.); Japan Science and Technology Agency Grant JPMJAL1607 (Y.I., T.B.); and Japan Society for the Promotion of Science Grants-in-Aid 17H06304 (Y.I., T.B.) and 18H01800 (T.B.). Manuscript received 4 April 2019 and in revised form 11 June 2019. Published, JLR Papers in Press, June 14, 2019 DOI https://doi.org/10.1194/jlr.D094516
PY - 2019
Y1 - 2019
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.
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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 -