Abstract Background and objective: Metabolomic research offers a deeper insight into biochemical changes in cancer metabolism and is a promising tool for identifying novel biomarkers. We aimed to evaluate the diagnostic and prognostic potential of metabolites in prostate cancer (PCa) tissue after radical prostatectomy. Material and methods: 107 matched-paired tissue samples collected after radical prostatectomy were subjected to the MxPTM Broad Profiling by gas chromatography- mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (Patent WO 2010/139711 A1: "Means and methods for diagnosing prostate carcinomas"). Aminoadipic acid, cerebronic acid, gluconic acid, glycerophosphoethanolamine, 2- hydroxybehenic acid, isopentenyl pyrophosphate, maltotriose, 7-methylguanine, and tricosanoic acid were related to clinicopathological variables like prostate volume, tumor stage, Gleason score, preoperative prostate-specific antigen (PSA), and disease recurrence in the follow-up. Non-parametric statistical tests, receiver- operating characteristics (ROC) and univariate and multivariate analyses (Kaplan- Meier curve; Cox regression) were performed. Results: All metabolites showed higher concentrations in malignant than in non- malignant samples except for gluconic acid and maltotriose, which had lower levels in tumors. ROC analyses showed a clear differentiation for all metabolites with a maximal area under the curve of 0.86 for tricosanoic acid. However, the metabolites were not related to tumor stage and Gleason grade. "Aminoadipic acid, gluconic acid, and maltotriose levels were associated with tumor recurrence (Kaplan-Meier analysis) and were, together with tumor stage and Gleason score, a successful metabolite combination in the multivariate Cox regression model for the prediction of tumor recurrence." Conclusions: This exemplary study performed with selected metabolites from a global metabolic profiling investigation proves that metabolites in prostate carcinoma tissue can be used, in combination with traditional pathological and histomorphological parameters, as promising diagnostic and prognostic tools. Citation Format: Glen Kristiansen, Regina Reszka, Beate Kamlage, Bianca Bethan, Michael Lein, Carsten Stephan, Klaus Jung. Metabolite profiling as tool for the identification of differentiating and prognostic markers of prostate carcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3224. doi:10.1158/1538-7445.AM2013-3224