title: Targeted Metabolomic Analysis of Head and Neck Cancer Cells Using High Performance Ion Chromatography Coupled with a Q Exactive HF Mass Spectrometer.
abstract: In this study, we have demonstrated a targeted metabolomics method for analysis of cancer cells, based on high-performance ion chromatography (IC) separation, Q Exactive HF MS for high-resolution and accurate-mass (HR/AM) measurement and the use of stable isotope-labeled internal standards for absolute quantitation. Our method offers great technical advantages for metabolite analysis, including exquisite sensitivity, high speed and reproducibility, and wide dynamic range. The high-performance IC provided fast separation of cellular metabolites within 20 min and excellent resolving power for polar molecules including many isobaric metabolites. The IC/Q Exactive HF MS achieved wide dynamic ranges of 5 orders of magnitude for six targeted metabolites, pyruvate, succinic acid, malic acid, citric acid, fumaric acid, and alpha-ketoglutaric acid, with R(2) ≈ 0.99. Using this platform, metabolites can be simultaneously quantified from low fmol/μL to nmol/μL levels in cellular samples. The high flow rate IC at 380 μL/min has shown excellent reproducibility for a large set of samples (150 injections), with minimal variations of retention time (SD < ± 0.03 min). In addition, the IC-MS-based approach acquires targeted and global metabolomic data in a same analytical run, and the use of stable isotope-labeled standards facilitates accurate quantitation of targeted metabolites in large-scale metabolomics analysis. This metabolomics approach has been successfully applied to analysis of targeted metabolites in head and neck cancer cells as well as cancer stem-like cells (CSCs), and the findings indicate that the metabolic phenotypes may be distinct between high and low invasive head and neck cancer cells and between CSCs and non-SCCs.
authors.affiliation: †School of Dentistry and Jonsson Comprehensive Cancer Center, University of California, Los Angeles, California 90095, United States.
authors.forename: Shen
authors.initials: S
authors.lastname: Hu
authors.affiliation: ‡Thermo Fisher Scientific Inc, San Jose, California 95134, United States.
authors.forename: Junhua
authors.initials: J
authors.lastname: Wang
authors.affiliation: †School of Dentistry and Jonsson Comprehensive Cancer Center, University of California, Los Angeles, California 90095, United States.
authors.forename: Eoon Hye
authors.initials: EH
authors.lastname: Ji
authors.affiliation: ‡Thermo Fisher Scientific Inc, San Jose, California 95134, United States.
authors.forename: Terri
authors.initials: T
authors.lastname: Christison
authors.affiliation: ‡Thermo Fisher Scientific Inc, San Jose, California 95134, United States.
authors.forename: Linda
authors.initials: L
authors.lastname: Lopez
authors.affiliation: ‡Thermo Fisher Scientific Inc, San Jose, California 95134, United States.
authors.forename: Yingying
authors.initials: Y
authors.lastname: Huang
mesh_terms: D002851:Chromatography, High Pressure Liquid; D019343:Citric Acid / Q000378:metabolism; D005650:Fumarates / Q000378:metabolism; D006258:Head and Neck Neoplasms / Q000378:metabolism* / Q000473:pathology*; D006801:Humans; D007656:Ketoglutaric Acids / Q000378:metabolism; D008293:Malates / Q000378:metabolism; D013058:Mass Spectrometry; D055432:Metabolomics; D019289:Pyruvic Acid / Q000378:metabolism; D019802:Succinic Acid / Q000378:metabolism
chemical_list: D005650:Fumarates; D007656:Ketoglutaric Acids; D008293:Malates; D019343:Citric Acid; C030298:malic acid; D019289:Pyruvic Acid; C032005:fumaric acid; D019802:Succinic Acid