Investigating the Effects of Evodia rutaecarpa on Caffeine-Infected Water

Caffeine is an emerging contaminant in waterways, raising concerns about oxidative stress, reproductive issues, and increased mortality in aquatic life. Due to caffeine’s water solubility, it stays within waterways for a long period of time creating hazards for aquatic wildlife. The available treatments for caffeine-polluted water all have drawbacks making large-scale implementation futile as well as necessitating preventative measures. Evodia rutaecarpa, also known as Wu Zhu Yu, is a Traditional Chinese Medicine (TCM) that has been used to combat the negative effects of caffeine. Evodiamine, a major constituent of Evodia rutaecarpa, was found to decrease sleep disruptions and erratic behaviors in caffeine-treated mice (Ko et. al., Biomol. Ther. 2018). Planarian worms, Girardia dorotocephala, are a species of flatworms we have used to study the protective effect of E. rutaecarpa on regeneration. In assays with caffeine-treated G. dorotocephala, we measured the rate of regeneration in order to find the small molecule from E. rutaecarpa responsible for caffeine protection, allowing for higher rates of survival and regeneration. Partitions of a commercially available tincture were made: ethyl acetate, chloroform, and the remaining water soluble tincture constituents not soluble in ethyl acetate and chloroform. Caffeine-laced water was treated with each partition and compared to the negative control group (no treatment) and the original tincture (positive control). It was found that ethyl acetate and chloroform partitions had the most stable growth of planaria with the lowest mortality rate. Ethyl acetate and chloroform partitions were then fractionated using silica gel to create four fractions per partition. These fractions are currently being tested. The known molecules will then be identified in the fractions by liquid chromatography mass spectrometry. Any novel compounds with bioactivity will be isolated and their structures determined. The most biologically active molecules could be used to reduce the negative effects of caffeine in waterways.