title: Determination of energy metabolites in cancer cells by porous graphitic carbon liquid chromatography electrospray ionization mass spectrometry for the assessment of energy metabolism.
abstract: A high performance liquid chromatography (HPLC) tandem mass spectrometric (MS/MS) method has been developed for the simultaneous determination of fifteen glucose, or acetate derived metabolites isolated from tumor cells. Glycolytic and tricarboxylic acid (TCA) cycle metabolites as well as acidic amino acids were separated on a HPLC porous graphitic carbon (PGC) column and simultaneously determined by means of triple quadrupole MS/MS using multiple reaction monitoring (MRM). Target compounds were eluted within 10 min with 8% v/v formic acid as an electronic modifier added to a 4:1 v/v methanol water mobile phase. The calibration is linear in the 1-100 μM concentration range for each analyte. The limit of detection ranges between 0.39 and 2.78 μM for the analytes concerned. To test the PGC-HPLC-MS/MS method in metabolomic studies, ZR-75.1 human mammary adenocarcinoma cells were labeled with U-(13)C glucose or 1-(13)C acetate. Applying the MRM mode, the incorporation of (13)C into metabolites, isolated from the tumor cells, and derived from glucose or acetate, could be properly identified.
authors.affiliation: Laboratory of Environmental Chemistry and Bioanalytics, Department of Analytical Chemistry, Institute of Chemistry, Eötvös Loránd University, Pázmány Péter stny 1/A, H-1117 Budapest, Hungary. Electronic address: szobosz@chem.elte.hu.
authors.forename: Norbert
authors.initials: N
authors.lastname: Szoboszlai
authors.affiliation: Institute of Analytical Chemistry and Food Chemistry, Graz University of Technology, Stremayrgasse 9, 8010 Graz, Austria.
authors.forename: Xinghua
authors.initials: X
authors.lastname: Guo
authors.affiliation: Research Centre for Natural Sciences of the Hungarian Academy of Sciences, Pusztaszeri u. 59-67, H-1025 Budapest, Hungary.
authors.forename: Olivér
authors.initials: O
authors.lastname: Ozohanics
authors.affiliation: 1st Institute of Pathology and Experimental Cancer Research, Semmelweis University, H-1085 Budapest, Hungary.
authors.forename: Júlia
authors.initials: J
authors.lastname: Oláh
authors.affiliation: Research Centre for Natural Sciences of the Hungarian Academy of Sciences, Pusztaszeri u. 59-67, H-1025 Budapest, Hungary.
authors.forename: Ágnes
authors.initials: Á
authors.lastname: Gömöry
authors.affiliation: Laboratory of Environmental Chemistry and Bioanalytics, Department of Analytical Chemistry, Institute of Chemistry, Eötvös Loránd University, Pázmány Péter stny 1/A, H-1117 Budapest, Hungary.
authors.forename: Victor G
authors.initials: VG
authors.lastname: Mihucz
authors.affiliation: 1st Institute of Pathology and Experimental Cancer Research, Semmelweis University, H-1085 Budapest, Hungary.
authors.forename: András
authors.initials: A
authors.lastname: Jeney
authors.affiliation: Research Centre for Natural Sciences of the Hungarian Academy of Sciences, Pusztaszeri u. 59-67, H-1025 Budapest, Hungary.
authors.forename: Károly
authors.initials: K
authors.lastname: Vékey
mesh_terms: D000230:Adenocarcinoma / Q000378:metabolism* / Q000473:pathology; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D002244:Carbon / Q000737:chemistry*; D002853:Chromatography, Liquid; D004734:Energy Metabolism; D006801:Humans; D010316:Particle Size; D016062:Porosity; D021241:Spectrometry, Mass, Electrospray Ionization; D013499:Surface Properties; D014407:Tumor Cells, Cultured
chemical_list: D002244:Carbon
keywords: Electrospray ionization; Intracellular; Tandem mass spectrometry; Targeted metabolomics; Triple quadrupole