Abstract:Non-alcoholic fatty liver disease is a pathology with a hard-to-detect onset and is estimated to be present in a quarter of the adult human population. To improve our understanding of the development of non-alcoholic fatty liver disease, we treated a human hepatoma cell line model, HepG2, with increasing concentrations of common fatty acids, namely myristic, palmitic and oleic acid. To reproduce more physiologically representative conditions, we also included combinations of these fatty acids and monitored the… Show more
“…Various lipidsoluble compounds, especially free fatty acids (FFAs), play significant roles in mammalian bodies and are involved in many diseases or disorders, such as cancers, fever, pain, and bipolar disorders. [9][10][11][12][13][14] However, their measurement in a single cell is not always easy due to the small amounts of these substances, the presence of isomers, and the complex sample preparation procedures.…”
Proton-transfer-reaction (PTR) mass spectrometry (MS), a widely used method for detecting tracelevels of volatile organic compounds in gaseous samples, can also be used for the analysis of small nonvolatile 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 200/90 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
“…Various lipidsoluble compounds, especially free fatty acids (FFAs), play significant roles in mammalian bodies and are involved in many diseases or disorders, such as cancers, fever, pain, and bipolar disorders. [9][10][11][12][13][14] However, their measurement in a single cell is not always easy due to the small amounts of these substances, the presence of isomers, and the complex sample preparation procedures.…”
Proton-transfer-reaction (PTR) mass spectrometry (MS), a widely used method for detecting tracelevels of volatile organic compounds in gaseous samples, can also be used for the analysis of small nonvolatile 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 200/90 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
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