{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T20:03:17Z","timestamp":1792180997399,"version":"4.4.0"},"reference-count":21,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2019,8,2]],"date-time":"2019-08-02T00:00:00Z","timestamp":1564704000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2019,8,2]],"date-time":"2019-08-02T00:00:00Z","timestamp":1564704000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Given the implications of oncometabolites [succinate, fumarate, and 2-hydroxyglutarate (2HG)] in cancer pathogenesis and therapeutics, quantitative determination of their tissue levels has significant diagnostic, prognostic, and therapeutic values. Here, we developed and validated a multiplex liquid chromatography coupled with tandem mass spectrometry (LC-MS\/MS) platform that allows simultaneous determination of oncometabolites (including succinate, fumarate and total 2HG) and other tricarboxylic acid cycle metabolites (\u03b1-ketoglutarate, malic acid, and glutamate) in frozen and FFPE tissues specimens. In addition, by employing chiral derivatization in the sample preparation, the platform enabled separation and determination of 2HG enantiomers (D- and L-2HG) in frozen and FFPE tissues. Isotope-labeled internal standard method was used for the quantitation. Linear calibration curve ranges in aqueous solution were 0.02\u201310, 0.2\u2013100, 0.002\u201310, and 0.002\u20135\u2009\u00b5M for succinate, fumarate, total 2HG, and D\/L-2HG, respectively. Intra- and inter-day precision and accuracy for individual oncometabolites were within the generally accepted criteria for bioanalytical method validation (&lt;15%). The recovery of spiked individual oncometabolites from pooled homogenate of FFPE or frozen tissue ranged 86\u2013112%. Method validation indicated the technical feasibility, reliability and reproducibility of the platform. Oncometabolites were notably lost during the routine FFPE process. The ratios of succinate to glutamate, fumarate to \u03b1-ketoglutarate, 2HG to glutamate, and D-2HG to L-2HG were reliable surrogate measurements for the detection of altered levels of oncometabolites in FFPE specimens. Our study laid a foundation for the utility of archival FFPE specimens for oncometabolite profiling as a valid technique in clinical research and routine medical care.<\/jats:p>","DOI":"10.1038\/s41598-019-47669-5","type":"journal-article","created":{"date-parts":[[2019,8,2]],"date-time":"2019-08-02T07:19:54Z","timestamp":1564730394000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Quantitative Profiling of Oncometabolites in Frozen and Formalin-Fixed Paraffin-Embedded Tissue Specimens by Liquid Chromatography Coupled with Tandem Mass Spectrometry"],"prefix":"10.1038","volume":"9","author":[{"given":"Xun","family":"Bao","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianmei","family":"Wu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian","family":"Shuch","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patricia","family":"LoRusso","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3255-0467","authenticated-orcid":false,"given":"Ranjit S.","family":"Bindra","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,8,2]]},"reference":[{"key":"47669_CR1","doi-asserted-by":"publisher","first-page":"85","DOI":"10.3389\/fonc.2012.00085","volume":"2","author":"M Yang","year":"2012","unstructured":"Yang, M., Soga, T., Pollard, P. J. & Adam, J. The emerging role of fumarate as an oncometabolite. Front Oncol 2, 85, https:\/\/doi.org\/10.3389\/fonc.2012.00085 (2012).","journal-title":"Front Oncol"},{"key":"47669_CR2","doi-asserted-by":"publisher","first-page":"813","DOI":"10.1056\/NEJMe0810213","volume":"360","author":"CB Thompson","year":"2009","unstructured":"Thompson, C. B. Metabolic enzymes as oncogenes or tumor suppressors. N Engl J Med 360, 813\u2013815, https:\/\/doi.org\/10.1056\/NEJMe0810213 (2009).","journal-title":"N Engl J Med"},{"key":"47669_CR3","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1016\/j.ccr.2010.01.020","volume":"17","author":"PS Ward","year":"2010","unstructured":"Ward, P. S. et al. The common feature of leukemia-associated IDH1 and IDH2 mutations is a neomorphic enzyme activity converting alpha-ketoglutarate to 2-hydroxyglutarate. Cancer Cell 17, 225\u2013234, https:\/\/doi.org\/10.1016\/j.ccr.2010.01.020 (2010).","journal-title":"Cancer Cell"},{"key":"47669_CR4","doi-asserted-by":"publisher","first-page":"3652","DOI":"10.1172\/JCI67228","volume":"123","author":"M Yang","year":"2013","unstructured":"Yang, M., Soga, T. & Pollard, P. J. Oncometabolites: linking altered metabolism with cancer. J Clin Invest 123, 3652\u20133658, https:\/\/doi.org\/10.1172\/JCI67228 (2013).","journal-title":"J Clin Invest"},{"key":"47669_CR5","doi-asserted-by":"publisher","first-page":"2231","DOI":"10.1093\/hmg\/ddi227","volume":"14","author":"PJ Pollard","year":"2005","unstructured":"Pollard, P. J. et al. Accumulation of Krebs cycle intermediates and over-expression of HIF1alpha in tumours which result from germline FH and SDH mutations. 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N Engl J Med 362, 369\u2013370, https:\/\/doi.org\/10.1056\/NEJMc0910063 (2010).","journal-title":"N Engl J Med"},{"key":"47669_CR8","doi-asserted-by":"publisher","first-page":"765","DOI":"10.1056\/NEJMoa0808710","volume":"360","author":"H Yan","year":"2009","unstructured":"Yan, H. et al. IDH1 and IDH2 mutations in gliomas. N Engl J Med 360, 765\u2013773, https:\/\/doi.org\/10.1056\/NEJMoa0808710 (2009).","journal-title":"N Engl J Med"},{"key":"47669_CR9","doi-asserted-by":"publisher","first-page":"291","DOI":"10.1016\/j.cmet.2015.06.021","volume":"22","author":"WM Oldham","year":"2015","unstructured":"Oldham, W. M., Clish, C. B., Yang, Y. & Loscalzo, J. Hypoxia-Mediated Increases in L-2-hydroxyglutarate Coordinate the Metabolic Response to Reductive Stress. Cell Metab 22, 291\u2013303, https:\/\/doi.org\/10.1016\/j.cmet.2015.06.021 (2015).","journal-title":"Cell Metab"},{"key":"47669_CR10","doi-asserted-by":"publisher","first-page":"1290","DOI":"10.1158\/2159-8290.CD-13-0696","volume":"4","author":"EH Shim","year":"2014","unstructured":"Shim, E. H. et al. L-2-Hydroxyglutarate: an epigenetic modifier and putative oncometabolite in renal cancer. Cancer Discov 4, 1290\u20131298, https:\/\/doi.org\/10.1158\/2159-8290.CD-13-0696 (2014).","journal-title":"Cancer Discov"},{"issue":"375","key":"47669_CR11","doi-asserted-by":"publisher","first-page":"eaal2463","DOI":"10.1126\/scitranslmed.aal2463","volume":"9","author":"Parker L. Sulkowski","year":"2017","unstructured":"Sulkowski, P. L. et al. 2-Hydroxyglutarate produced by neomorphic IDH mutations suppresses homologous recombination and induces PARP inhibitor sensitivity. 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Brain Pathol 22, 26\u201331, https:\/\/doi.org\/10.1111\/j.1750-3639.2011.00506.x (2012).","journal-title":"Brain Pathol"},{"key":"47669_CR16","doi-asserted-by":"publisher","first-page":"1391","DOI":"10.1373\/clinchem.2004.033399","volume":"50","author":"EA Struys","year":"2004","unstructured":"Struys, E. A., Jansen, E. E., Verhoeven, N. M. & Jakobs, C. Measurement of urinary D- and L-2-hydroxyglutarate enantiomers by stable-isotope-dilution liquid chromatography-tandem mass spectrometry after derivatization with diacetyl-L-tartaric anhydride. Clin Chem 50, 1391\u20131395, https:\/\/doi.org\/10.1373\/clinchem.2004.033399 (2004).","journal-title":"Clin Chem"},{"key":"47669_CR17","doi-asserted-by":"publisher","DOI":"10.1038\/srep15217","volume":"5","author":"QY Cheng","year":"2015","unstructured":"Cheng, Q. Y. et al. 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