Targeted Metabolomic Analysis of Head and Neck Cancer Cells Using High Performance Ion Chromatography Coupled with a Q Exactive HF Mass Spectrometer

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• 作者：Shen Hu ; Junhua Wang ; Eoon Hye Ji ; Terri Christison ; Linda Lopez ; Yingying Huang
• 刊名：Analytical Chemistry
• 出版年：2015
• 出版时间：June 16, 2015
• 年：2015
• 卷：87
• 期：12
• 页码：6371-6379
• 全文大小：554K
• ISSN：1520-6882

文摘

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 R² 鈮?0.99. Using this platform, metabolites can be simultaneously quantified from low fmol/渭L to nmol/渭L levels in cellular samples. The high flow rate IC at 380 渭L/min has shown excellent reproducibility for a large set of samples (150 injections), with minimal variations of retention time (SD < 卤 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.