Attempts to Detect Lipid Metabolites from a Single Cell Using Proton-Transfer-Reaction Mass Spectrometry Coupled with Micro-Scale Supercritical Fluid Extraction: A Preliminary Study

Toshinobu Hondo; Chihiro Ota; Kohta Nakatani; Yumi Miyake; Hiroshi Furutani; Takeshi Bamba; Michisato Toyoda

doi:10.5702/massspectrometry.A0112

Attempts to Detect Lipid Metabolites from a Single Cell Using Proton-Transfer-Reaction Mass Spectrometry Coupled with Micro-Scale Supercritical Fluid Extraction: A Preliminary Study

Mass Spectrom (Tokyo). 2022;11(1):A0112. doi: 10.5702/massspectrometry.A0112. Epub 2022 Dec 29.

Authors

Toshinobu Hondo^{1

2}, Chihiro Ota^{2

3}, Kohta Nakatani⁴, Yumi Miyake², Hiroshi Furutani^{2

5}, Takeshi Bamba⁴, Michisato Toyoda²

Affiliations

¹ MS-Cheminformatics LLC, 2-13-21 Sasao-nishi, Toin, Inabe, Mie 511-0231, Japan.
² Forefront Research Center, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, Japan.
³ Graduate School of Science and Engineering, Kansai University, 3-3-35 Yamate-cho, Suita, Osaka 564-8680, Japan.
⁴ Division of Metabolomics, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.
⁵ Center for Scientific Instrument Renovation and Manufacturing Support, Osaka University, 1-2 Machikaneyama, Toyonaka, Osaka 560-0043, Japan.

Abstract

Proton-transfer-reaction (PTR) mass spectrometry (MS), a widely used method for detecting trace-levels of volatile organic compounds in gaseous samples, can also be used for the analysis of small non-volatile molecules by using supercritical fluid as a transporter for the molecules. Supercritical fluid extraction (SFE) is a method that permits lipophilic compounds to be rapidly and selectively extracted from complex matrices. The combination of the high sensitivity of PTR MS with the SFE is a potentially novel method for analyzing small molecules in a single cell, particularly for the analysis of lipophilic compounds. We preliminarily evaluated this method for analyzing the components of a single HeLa cell that is fixed on a stainless steel frit and is then directly introduces the SFE extracts into the PTR MS. A total of 200/91 ions were observed in positive/negative ion mode time-of-flight mass spectra, and the masses of 11/10 ions could be matched to chemical formulae obtained from the LipidMaps lipids structure database. Using various authentic lipophilic samples, the method could be used to detect free fatty acids in the sub-femtomole to femtomole order in the negative ion mode, the femtomole to sub-picomole order for fat-soluble vitamins, and the picomole order for poly aromatic hydrocarbons in both the positive and negative ion mode.

Keywords: proton-transfer-reaction mass spectrometry; single-cell lipid metabolite analysis; supercritical fluid chromatography; supercritical fluid extraction; water ion chemical ionization.