ACS Publications. Most Trusted. Most Cited. Most Read
My Activity

Figure 1Loading Img

Stable Isotope- and Mass Spectrometry-based Metabolomics as Tools in Drug Metabolism: A Study Expanding Tempol Pharmacology

View Author Information
Laboratory of Metabolism, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, United States
# Department of Food Science and Nutrition, University of Minnesota, St. Paul, Minnesota 55108, United States
§ Radiation Biology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, United States
Department of Veterinary and Biomedical Sciences and the Center for Molecular Toxicology and Carcinogenesis, The Pennsylvania State University, University Park, Pennsylvania 16802, United States
*Tel: (814) 867-4565. Fax: (814) 863-1696. E-mail: [email protected]
Cite this: J. Proteome Res. 2013, 12, 3, 1369–1376
Publication Date (Web):January 10, 2013
Copyright © 2013 American Chemical Society

    Article Views





    Other access options
    Supporting Info (1)»


    Abstract Image

    The application of mass spectrometry-based metabolomics in the field of drug metabolism has yielded important insights not only into the metabolic routes of drugs but has provided unbiased, global perspectives of the endogenous metabolome that can be useful for identifying biomarkers associated with mechanism of action, efficacy, and toxicity. In this report, a stable isotope- and mass spectrometry-based metabolomics approach that captures both drug metabolism and changes in the endogenous metabolome in a single experiment is described. Here the antioxidant drug tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl) was chosen because its mechanism of action is not completely understood and its metabolic fate has not been studied extensively. Furthermore, its small size (MW = 172.2) and chemical composition (C9H18NO2) make it challenging to distinguish from endogenous metabolites. In this study, mice were dosed with tempol or deuterated tempol (C9D17HNO2) and their urine was profiled using ultraperformance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. Principal component analysis of the urinary metabolomics data generated a Y-shaped scatter plot containing drug metabolites (protonated and deuterated) that were clearly distinct from the endogenous metabolites. Ten tempol drug metabolites, including eight novel metabolites, were identified. Phase II metabolism was the major metabolic pathway of tempol in vivo, including glucuronidation and glucosidation. Urinary endogenous metabolites significantly elevated by tempol treatment included 2,8-dihydroxyquinoline (8.0-fold, P < 0.05) and 2,8-dihydroxyquinoline-β-d-glucuronide (6.8-fold, P < 0.05). Urinary endogenous metabolites significantly attenuated by tempol treatment including pantothenic acid (1.3-fold, P < 0.05) and isobutrylcarnitine (5.3-fold, P < 0.01). This study underscores the power of a stable isotope- and mass spectrometry-based metabolomics in expanding the view of drug pharmacology.

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.


    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. You can change your affiliated institution below.

    Supporting Information

    Jump To

    Supplemental metabolites identification and figures. This material is available free of charge via the Internet at

    Terms & Conditions

    Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system:

    Cited By

    This article is cited by 28 publications.

    1. Yuan Tian, Wei Gui, Philip B. Smith, Imhoi Koo, Iain A. Murray, Margherita T. Cantorna, Gary H. Perdew, Andrew D. Patterson. Isolation and Identification of Aryl Hydrocarbon Receptor Modulators in White Button Mushrooms (Agaricus bisporus). Journal of Agricultural and Food Chemistry 2019, 67 (33) , 9286-9294.
    2. Troy D. Hubbard, Qing Liu, Iain A. Murray, Fangcong Dong, Charles Miller, III, Philip B. Smith, Krishne Gowda, Jyh Ming Lin, Shantu Amin, Andrew D. Patterson, Gary H. Perdew. Microbiota Metabolism Promotes Synthesis of the Human Ah Receptor Agonist 2,8-Dihydroxyquinoline. Journal of Proteome Research 2019, 18 (4) , 1715-1724.
    3. Robert G. Nichols, Nicole E. Hume, Philip B. Smith, Jeffrey M. Peters, and Andrew D. Patterson . Omics Approaches To Probe Microbiota and Drug Metabolism Interactions. Chemical Research in Toxicology 2016, 29 (12) , 1987-1997.
    4. Fei Li, Xiu-Wei Yang, Kristopher W. Krausz, Robert G. Nichols, Wei Xu, Andrew D. Patterson, and Frank J. Gonzalez . Modulation of Colon Cancer by Nutmeg. Journal of Proteome Research 2015, 14 (4) , 1937-1946.
    5. Bernhard Kluger, Christoph Bueschl, Nora Neumann, Romana Stückler, Maria Doppler, Alexander W. Chassy, Andrew L. Waterhouse, Justyna Rechthaler, Niklas Kampleitner, Gerhard G. Thallinger, Gerhard Adam, Rudolf Krska, and Rainer Schuhmacher . Untargeted Profiling of Tracer-Derived Metabolites Using Stable Isotopic Labeling and Fast Polarity-Switching LC–ESI-HRMS. Analytical Chemistry 2014, 86 (23) , 11533-11537.
    6. Fei Li, Changtao Jiang, Michele C. Larsen, Justin Bushkofsky, Kristopher W. Krausz, Ting Wang, Colin R. Jefcoate, and Frank J. Gonzalez . Lipidomics Reveals a Link between CYP1B1 and SCD1 in Promoting Obesity. Journal of Proteome Research 2014, 13 (5) , 2679-2687.
    7. Pavan K. Kancharla, Takayuki Kato, and David Crich . Probing the Influence of Protecting Groups on the Anomeric Equilibrium in Sialic Acid Glycosides with the Persistent Radical Effect. Journal of the American Chemical Society 2014, 136 (14) , 5472-5480.
    8. Pengyi Chen, Chengguo Pang, Tianen Xu, Peiliang Dong, Hua Han. Characterization of chemical constituents and metabolites in vivo and in vitro after oral administration of Wuteng tablets in rats by UHPLC–Q/TOF–MS. Biomedical Chromatography 2023, 37 (10)
    9. Yuanfu Lu, Xue-Mei Zhao, Zhaoyong Hu, Li Wang, Feng Li. LC–MS-Based Metabolomics in the Study of Drug-Induced Liver Injury. Current Pharmacology Reports 2019, 5 (1) , 56-67.
    10. Jingwei Cai, Robert G. Nichols, Imhoi Koo, Zachary A. Kalikow, Limin Zhang, Yuan Tian, Jingtao Zhang, Philip B. Smith, Andrew D. Patterson, . Multiplatform Physiologic and Metabolic Phenotyping Reveals Microbial Toxicity. mSystems 2018, 3 (6)
    11. Diren Beyoğlu, Yuyin Zhou, Chi Chen, Jeffrey R. Idle. Mass isotopomer-guided decluttering of metabolomic data to visualize endogenous biomarkers of drug toxicity. Biochemical Pharmacology 2018, 156 , 491-500.
    12. Qi Zhao, Jia-Le Zhang, Fei Li. Application of Metabolomics in the Study of Natural Products. Natural Products and Bioprospecting 2018, 8 (4) , 321-334.
    13. Dan-Dan Hu, Xing-Long Chen, Xue-Rong Xiao, Yi-Kun Wang, Fang Liu, Qi Zhao, Xian Li, Xiu-Wei Yang, Fei Li. Comparative metabolism of tripolide and triptonide using metabolomics. Food and Chemical Toxicology 2018, 115 , 98-108.
    14. Qi Zhao, Xin-Mei Li, Hong-Ning Liu, Frank J. Gonzalez, Fei Li. Metabolic map of osthole and its effect on lipids. Xenobiotica 2018, 48 (3) , 285-299.
    15. Qian-Qian Lv, Xiao-Nan Yang, Dong-Mei Yan, Wei-Qing Liang, Hong-Ning Liu, Xiu-Wei Yang, Fei Li. Metabolic profiling of dehydrodiisoeugenol using xenobiotic metabolomics. Journal of Pharmaceutical and Biomedical Analysis 2017, 145 , 725-733.
    16. Xiaoli Wei, Biyun Shi, Imhoi Koo, Xinmin Yin, Pawel Lorkiewicz, Hamid Suhail, Ramandeep Rattan, Shailendra Giri, Craig J. McClain, Xiang Zhang. Analysis of stable isotope assisted metabolomics data acquired by GC-MS. Analytica Chimica Acta 2017, 980 , 25-32.
    17. Wei Jia, Xiaogang Chu, James Chang, Perry G. Wang, Ying Chen, Feng Zhang. High-throughput untargeted screening of veterinary drug residues and metabolites in tilapia using high resolution orbitrap mass spectrometry. Analytica Chimica Acta 2017, 957 , 29-39.
    18. Yuwei Lu, Chi Chen. Metabolomics: Bridging Chemistry and Biology in Drug Discovery and Development. Current Pharmacology Reports 2017, 3 (1) , 16-25.
    19. Frank J. Gonzalez, Changtao Jiang, Andrew D. Patterson. An Intestinal Microbiota–Farnesoid X Receptor Axis Modulates Metabolic Disease. Gastroenterology 2016, 151 (5) , 845-859.
    20. Ningli Wang, Jianteng Wei, Yewei Liu, Dong Pei, Qingping Hu, Yu Wang, Duolong Di. Discovery of biomarkers for oxidative stress based on cellular metabolomics. Biomarkers 2016, 21 (5) , 449-457.
    21. Julian R Marchesi, David H Adams, Francesca Fava, Gerben D A Hermes, Gideon M Hirschfield, Georgina Hold, Mohammed Nabil Quraishi, James Kinross, Hauke Smidt, Kieran M Tuohy, Linda V Thomas, Erwin G Zoetendal, Ailsa Hart. The gut microbiota and host health: a new clinical frontier. Gut 2016, 65 (2) , 330-339.
    22. Frank J Gonzalez, Zhong-Ze Fang, Xiaochao Ma. Transgenic mice and metabolomics for study of hepatic xenobiotic metabolism and toxicity. Expert Opinion on Drug Metabolism & Toxicology 2015, 11 (6) , 869-881.
    23. Amélie Desmarais, Estelle Pujos‐Guillot, Bernard Lyan, Jean‐François Martin, Nadine Leblanc, Paul Angers, Jean‐Louis Sébédio. Cyclic Fatty Acids Found in Frying Oils are Detoxified via Classical Drug Metabolic Pathway but also by β‐Oxidation and Eliminated as Conjugates in Rats. Lipids 2015, 50 (4) , 381-396.
    24. Mar Garcia-Aloy, Rafael Llorach, Mireia Urpi-Sarda, Sara Tulipani, Jordi Salas-Salvadó, Miguel Angel Martínez-González, Dolores Corella, Montserrat Fitó, Ramon Estruch, Lluis Serra-Majem, Cristina Andres-Lacueva. Nutrimetabolomics fingerprinting to identify biomarkers of bread exposure in a free-living population from the PREDIMED study cohort. Metabolomics 2015, 11 (1) , 155-165.
    25. M. Ferrara. Future perspectives for metabolomics in nutrition research. 2015, 231-236.
    26. Zhong-Ze Fang, Frank J. Gonzalez. LC–MS-based metabolomics: an update. Archives of Toxicology 2014, 88 (8) , 1491-1502.
    27. . Modern methodologies and tools for human hazard assessment of chemicals. EFSA Journal 2014
    28. Fei Li, Changtao Jiang, Kristopher W. Krausz, Yunfei Li, Istvan Albert, Haiping Hao, Kristin M. Fabre, James B. Mitchell, Andrew D. Patterson, Frank J. Gonzalez. Microbiome remodelling leads to inhibition of intestinal farnesoid X receptor signalling and decreased obesity. Nature Communications 2013, 4 (1)

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    You’ve supercharged your research process with ACS and Mendeley!

    STEP 1:
    Click to create an ACS ID

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Your Mendeley pairing has expired. Please reconnect