title: High-Throughput Indirect Quantitation of13C Enriched Metabolites Using1H NMR.
abstract: Nuclear magnetic resonance (NMR) spectroscopy is widely used in metabolomics to perform quantitative profiling of low-molecular weight compounds from biological specimens. The measurement of endogenous metabolites using NMR has proven to be a powerful tool to identify new metabolic biomarkers in physiological and pathological conditions, and to study and evaluate treatment efficiency. In this study we present a rapid approach to indirectly quantify13C enriched molecules using one-dimensional (1D)1H NMR. We demonstrate this approach using isotopically labeled [1,6-13C]glucose and in four different cell lines. We confirm the applicability of this approach for treatment follow-up, utilizing a renal cancer cell line with rapamycin as a tool compound to study changes in metabolic profiles. Finally, we validate the applicability of this method to study metabolic biomarkers from ex vivo tumor extracts, after infusion, using isotopically enriched glucose. Given the high throughput and increased sensitivity of direct-detect1H NMR, this analytical approach provides an avenue for simple and rapid metabolic analysis of biological samples including blood, urine, and biopsies.
authors.affiliation: Radiology and Molecular Pharmacology Program, Memorial Sloan Kettering Cancer Center , New York, New York 10065, United Sates.
authors.forename: Valentina
authors.initials: V
authors.lastname: Di Gialleonardo
authors.affiliation: Radiology and Molecular Pharmacology Program, Memorial Sloan Kettering Cancer Center , New York, New York 10065, United Sates.
authors.forename: Sui Seng
authors.initials: SS
authors.lastname: Tee
authors.affiliation: Radiology and Molecular Pharmacology Program, Memorial Sloan Kettering Cancer Center , New York, New York 10065, United Sates.
authors.forename: Hannah N
authors.initials: HN
authors.lastname: Aldeborgh
authors.affiliation: Radiology and Molecular Pharmacology Program, Memorial Sloan Kettering Cancer Center , New York, New York 10065, United Sates.
authors.forename: Vesselin Z
authors.initials: VZ
authors.lastname: Miloushev
authors.affiliation: Radiology and Molecular Pharmacology Program, Memorial Sloan Kettering Cancer Center , New York, New York 10065, United Sates.
authors.forename: Lidia S
authors.initials: LS
authors.lastname: Cunha
authors.affiliation: Radiology and Molecular Pharmacology Program, Memorial Sloan Kettering Cancer Center , New York, New York 10065, United Sates.
authors.forename: George D
authors.initials: GD
authors.lastname: Sukenick
authors.affiliation: Radiology and Molecular Pharmacology Program, Memorial Sloan Kettering Cancer Center , New York, New York 10065, United Sates.|Weill Cornell Medical College, New York, New York 10065, United States.
authors.forename: Kayvan R
authors.initials: KR
authors.lastname: Keshari
mesh_terms: D002247:Carbon Isotopes; D002460:Cell Line; D005947:Glucose / Q000737:chemistry; D057166:High-Throughput Screening Assays; D006801:Humans; D055432:Metabolomics; D015394:Molecular Structure; D066244:Proton Magnetic Resonance Spectroscopy; D015398:Signal Transduction / Q000187:drug effects; D058570:TOR Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D002247:Carbon Isotopes; D058570:TOR Serine-Threonine Kinases; D005947:Glucose