To address the goals of developing predictive biomarkers of adverse reactions to radiation treatment, we propose to perform a multi-site analytic validation of the product (metabolite panel kit) developed in the Phase I SBIR effort. Results of plasma metabolite analysis in defined cohorts of patients treated with SBRT for prostate cancer will be tested for precision , accuracy and reproducibility in three independent, geographically distributed testing laboratories. Clinical samples obtained from patients with prostate cancers and samples spiked with known quantities of metabolites will be distributed for analytic validation of Shuttle's metabolite kit. The data derived in the Phase I effort have defined metabolite panels for potential use as predictive biomarkers of radiation late effects. here, we propose three technical objectives: 1. To manufacture 500 metabolite panel kits and analysis software 2. Test and validate the performance of kits and software at three independent sites and 3. With the assistance of regulatory consultant and FDA input, design an expanded multi institutional validation clinical trial to test the metabolite panel kit for use as a potential metabolite biomarker for clinical application. Demonstration of proof of principle in patients with prostate cancer may support development and commercialization of the metabolite biomarker product.