Gumiganghwal-tang (GT) is a traditional herbal prescription widely used to treat inflammatory diseases in Asia. In this study, we evaluated neuroprotective effect and acetylcholinesterase (AChE) inhibitory activity of Gland compared with fermented GT (FGT). In order to better understand the neuroprotective mechanism, intracellular reactive oxygen species (ROS) production was investigated and high-performance liquid chromatography with diode-array detection (HPLC-DAD) analysis of contents of GT and FGT were conducted. As a result, FGT showed more potent protective effect against glutamate-induced HT22 cell death than GT and inhibited AChE activity. The neuroprotective effect of FGT is associated with inhibition of ROS production. The HPLC-DAD analytical results indicated that FGT contains higher content of bergapten and atractylenolide III than that of GT. In conclusion, FGT have neuroprotective effect and AChE inhibition, and may be useful source for treatment of neurodegenerative disease. (C) 2014, The Society for Biotechnology, Japan. All rights reserved.