Comparison of simple monophasic versus classical biphasic extraction protocols for comprehensive UHPLC-MS/MS lipidomic analysis of Hela cells
| Autor: | Corinna Sanwald, Carlos Calderón, Jörg Schlotterbeck, Bernhard Drotleff, Michael Lämmerhofer |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Methyl Ethers
Negative mode 02 engineering and technology Mass spectrometry 01 natural sciences Biochemistry Uhplc ms ms Analytical Chemistry 2-Propanol HeLa Tandem Mass Spectrometry Lipidomics Humans Environmental Chemistry Lipid profiling Data-independent acquisition Chromatography High Pressure Liquid Spectroscopy Chromatography biology Chemistry Methanol Solid Phase Extraction 010401 analytical chemistry Extraction (chemistry) Water 021001 nanoscience & nanotechnology biology.organism_classification Lipids 0104 chemical sciences Solvents Chloroform 0210 nano-technology HeLa Cells |
| Zdroj: | Analytica Chimica Acta. 1048:66-74 |
| ISSN: | 0003-2670 |
| DOI: | 10.1016/j.aca.2018.10.035 |
| Popis: | In this study, two monophasic isopropanol-water mixtures (IPA:H2O 75:25 v/v and IPA:H2O 90:10 v/v) were compared with traditionally employed biphasic methods of Bligh & Dyer and Matyash et al. as extraction systems for lipidomics analysis in Hela cells. Samples were analyzed by UHPLC-ESI-QTOF-MS/MS in positive and negative mode using sequential window acquisition of all theoretical fragment ion spectra (SWATH) and a relatively new software (MS-DIAL) was employed for the processing of the data which includes detection of peaks, MS/MS spectra deconvolution, identification of detected lipids and alignment of peaks through the analyzed samples. The studied performance parameters such as precision, recoveries of isotopically labeled internal standards and endogenous lipids, number of extracted lipids, and complexity of employed procedure showed that extraction with IPA:H2O 90:10 v/v performs similar to the Matyash protocol and better than Bligh & Dyer as well as IPA:H2O 75:25 v/v. However, less complex monophasic protocol which is simpler to implement and can be executed in plastic rather than glass, make the monophasic IPA:H2O 90:10 v/v protocol an excellent alternative to the classical biphasic protocols for reversed phase LC-MS lipidomics studies. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect