Online Direct Infusion Mass Spectrometry of Liquid–Liquid Extraction Phases for Metabolite and Lipid Profiling with the Direct Infusion Probe
<b>Background/Objectives:</b> Profiling of metabolites and lipids in biological samples can provide invaluable insights into life-sustaining chemical processes. The ability to detect both metabolites and lipids in the same sample can enhance these understandings and connect cellular dyna...
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2024-10-01
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| author | Cátia Marques Lena Blaase Ingela Lanekoff |
| author_facet | Cátia Marques Lena Blaase Ingela Lanekoff |
| author_sort | Cátia Marques |
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| description | <b>Background/Objectives:</b> Profiling of metabolites and lipids in biological samples can provide invaluable insights into life-sustaining chemical processes. The ability to detect both metabolites and lipids in the same sample can enhance these understandings and connect cellular dynamics. However, simultaneous detection of metabolites and lipids is generally hampered by chromatographic systems tailored to one molecular type. This void can be filled by direct infusion mass spectrometry (MS), where all ionizable molecules can be detected simultaneously. However, in direct infusion MS, the high chemical complexity of biological samples can introduce limitations in detectability due to matrix effects causing ionization suppression. <b>Methods:</b> Decreased sample complexity and increased detectability and molecular coverage was provided by combining our direct infusion probe (DIP) with liquid–liquid extraction (LLE) and directly sampling the different phases for direct infusion. Three commonly used LLE methods for separating lipids and metabolites were evaluated. <b>Results:</b> The butanol–methanol (BUME) method was found to be preferred since it provides high molecular coverage and have low solvent toxicity. The established BUME DIP-MS method was used as a fast and sensitive analysis tool to study chemical changes in insulin-secreting cells upon glucose stimulation. By analyzing the metabolome at distinct time points, down to 1-min apart, we found high dynamics of the intracellular metabolome. <b>Conclusions:</b> The rapid workflow with LLE DIP-MS enables higher sensitivity of phase separated metabolites and lipids. The application of BUME DIP-MS provides novel information on the dynamics of the intracellular metabolome of INS-1 during the two phases of insulin release for both metabolite and lipid classes. |
| format | Article |
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| institution | Kabale University |
| issn | 2218-1989 |
| language | English |
| publishDate | 2024-10-01 |
| publisher | MDPI AG |
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| series | Metabolites |
| spelling | doaj-art-be1aa3c43a2f4fa4a0d167af651631a32024-11-26T18:13:04ZengMDPI AGMetabolites2218-19892024-10-01141158710.3390/metabo14110587Online Direct Infusion Mass Spectrometry of Liquid–Liquid Extraction Phases for Metabolite and Lipid Profiling with the Direct Infusion ProbeCátia Marques0Lena Blaase1Ingela Lanekoff2Department of Chemistry—BMC, Uppsala University, Husargatan 3, 75 123 Uppsala, SwedenDepartment of Chemistry—BMC, Uppsala University, Husargatan 3, 75 123 Uppsala, SwedenDepartment of Chemistry—BMC, Uppsala University, Husargatan 3, 75 123 Uppsala, Sweden<b>Background/Objectives:</b> Profiling of metabolites and lipids in biological samples can provide invaluable insights into life-sustaining chemical processes. The ability to detect both metabolites and lipids in the same sample can enhance these understandings and connect cellular dynamics. However, simultaneous detection of metabolites and lipids is generally hampered by chromatographic systems tailored to one molecular type. This void can be filled by direct infusion mass spectrometry (MS), where all ionizable molecules can be detected simultaneously. However, in direct infusion MS, the high chemical complexity of biological samples can introduce limitations in detectability due to matrix effects causing ionization suppression. <b>Methods:</b> Decreased sample complexity and increased detectability and molecular coverage was provided by combining our direct infusion probe (DIP) with liquid–liquid extraction (LLE) and directly sampling the different phases for direct infusion. Three commonly used LLE methods for separating lipids and metabolites were evaluated. <b>Results:</b> The butanol–methanol (BUME) method was found to be preferred since it provides high molecular coverage and have low solvent toxicity. The established BUME DIP-MS method was used as a fast and sensitive analysis tool to study chemical changes in insulin-secreting cells upon glucose stimulation. By analyzing the metabolome at distinct time points, down to 1-min apart, we found high dynamics of the intracellular metabolome. <b>Conclusions:</b> The rapid workflow with LLE DIP-MS enables higher sensitivity of phase separated metabolites and lipids. The application of BUME DIP-MS provides novel information on the dynamics of the intracellular metabolome of INS-1 during the two phases of insulin release for both metabolite and lipid classes.https://www.mdpi.com/2218-1989/14/11/587direct infusion probeliquid–liquid extractionglucose stimulationINS-1 cellstime-dependent analysishigh-resolution mass spectrometry |
| spellingShingle | Cátia Marques Lena Blaase Ingela Lanekoff Online Direct Infusion Mass Spectrometry of Liquid–Liquid Extraction Phases for Metabolite and Lipid Profiling with the Direct Infusion Probe Metabolites direct infusion probe liquid–liquid extraction glucose stimulation INS-1 cells time-dependent analysis high-resolution mass spectrometry |
| title | Online Direct Infusion Mass Spectrometry of Liquid–Liquid Extraction Phases for Metabolite and Lipid Profiling with the Direct Infusion Probe |
| title_full | Online Direct Infusion Mass Spectrometry of Liquid–Liquid Extraction Phases for Metabolite and Lipid Profiling with the Direct Infusion Probe |
| title_fullStr | Online Direct Infusion Mass Spectrometry of Liquid–Liquid Extraction Phases for Metabolite and Lipid Profiling with the Direct Infusion Probe |
| title_full_unstemmed | Online Direct Infusion Mass Spectrometry of Liquid–Liquid Extraction Phases for Metabolite and Lipid Profiling with the Direct Infusion Probe |
| title_short | Online Direct Infusion Mass Spectrometry of Liquid–Liquid Extraction Phases for Metabolite and Lipid Profiling with the Direct Infusion Probe |
| title_sort | online direct infusion mass spectrometry of liquid liquid extraction phases for metabolite and lipid profiling with the direct infusion probe |
| topic | direct infusion probe liquid–liquid extraction glucose stimulation INS-1 cells time-dependent analysis high-resolution mass spectrometry |
| url | https://www.mdpi.com/2218-1989/14/11/587 |
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