title: DAPNe with micro-capillary separatory chemistry-coupled to MALDI-MS for the analysis of polar and non-polar lipid metabolism in one cell.
abstract: The cellular metabolome is considered to be a representation of cellular phenotype and cellular response to changes to internal or external events. Methods to expand the coverage of the expansive physiochemical properties that makeup the metabolome currently utilize multi-step extractions and chromatographic separations prior to chemical detection, leading to lengthy analysis times. In this study, a single-step procedure for the extraction and separation of a sample using a micro-capillary as a separatory funnel to achieve analyte partitioning within an organic/aqueous immiscible solvent system is described. The separated analytes are then spotted for MALDI-MS imaging and distribution ratios are calculated. Initially, the method is applied to standard mixtures for proof of partitioning. The extraction of an individual cell is non-reproducible; therefore, a broad chemical analysis of metabolites is necessary and will be illustrated with the one-cell analysis of a single Snu-5 gastric cancer cell taken from a cellular suspension. The method presented here shows a broad partitioning dynamic range as a single-step method for lipid analysis demonstrating a decrease in ion suppression often present in MALDI analysis of lipids. Graphical Abstract ᅟ.
authors.affiliation: Department of Chemistry, University of North Texas, Denton, TX, USA.
authors.forename: Jason S
authors.initials: JS
authors.lastname: Hamilton
authors.affiliation: Department of Chemistry, University of North Texas, Denton, TX, USA.
authors.forename: Roberto
authors.initials: R
authors.lastname: Aguilar
authors.affiliation: Department of Chemistry, Texas Women's Univeristy, Denton, TX, USA.
authors.forename: Robby A
authors.initials: RA
authors.lastname: Petros
authors.affiliation: Department of Chemistry, University of North Texas, Denton, TX, USA. gverbeck@unt.edu.
authors.forename: Guido F
authors.initials: GF
authors.lastname: Verbeck
mesh_terms: D045744:Cell Line, Tumor; D005591:Chemical Fractionation / Q000295:instrumentation*; D004867:Equipment Design; D006801:Humans; D050356:Lipid Metabolism; D055442:Metabolome; D055432:Metabolomics / Q000295:instrumentation*; D059010:Single-Cell Analysis / Q000295:instrumentation*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation*; D013274:Stomach Neoplasms / Q000378:metabolism
keywords: DAPNe; MALDI; Nanomanipulation; One-cell