The aim of the present study was to prepare Tc-99m-folate-glucaric acid and investigate the radiopharmaceutical potential for tumors imaging that over express folate receptor. Folate-glucaric acid was synthesized and the synthesized folate conjugate was confirmed with H-1 NMR and LC-MS/MS methods. Folate-glucaric acid was labeled with Tc-99m, and its radiolabelling efficiency was found as 96 +/- 2.0%. Biodistribution study of Tc-99m-folate-glucaric acid was carried out in vivo using two groups of female Albino Wistar rats: folate receptor (FR) saturated and unsaturated. Biodistribution study showed that Tc-99m-folate-glucaric acid indicated high uptake in folate receptor rich tissues such as breast, ovary and uterus. Therefore, Tc-99m-folate-glucaric acid shows good radiolabeling and biodistribution in FR organs, the radiolabeled conjugate is a reason potentially useful radiopharmaceutical for detection of FR-positive tumors. (C) 2011 Elsevier B.V. All rights reserved.