Characterization of Phenethyl Cinnamamide Compounds from Hemp Seed and Determination of Their Melanogenesis Inhibitory Activity.

Hyperpigmentation is induced by the overactivation of tyrosinase, which is a rate-limiting enzyme in melanogenesis. The defatted extract of hemp (Cannabis sativa L.) seed is known to have inhibitory effects on melanogenesis; however, effective compounds in the extract have not been identified yet. In this study, three phenethyl cinnamamides present in hemp seed extract were prepared by purification and chemical synthesis and were assessed for their inhibitory effect on melanogenesis in B16F10 melanoma cells. A comparison of the anti-melanogenesis and anti-tyrosinase activity of hemp seed solvent fractions revealed that the ethyl acetate fraction possessed the greatest potential for suppressing melanogenesis in melanoma cells by decreasing tyrosinase activity. We tentatively identified 26 compounds in the ethyl acetate fraction by comparing spectroscopic data with the literature. Three phenethyl cinnamamides such as N-trans-caffeoyltyramine, N-trans-coumaroyltyramine, and N-trans-feruloyltyramine present abundantly in the ethyl acetate fraction were prepared and their anti-melanogenesis and anti-tyrosinase activities in melanoma cells were evaluated. We found that N-trans-caffeoyltyramine and N-trans-feruloyltyramine inhibited alpha melanocyte stimulating hormone (α-MSH)-induced melanogenesis without cytotoxicity, while N-trans-coumaroyltyramine inhibited melanogenesis with cytotoxicity. IC50 values of N-trans-caffeoyltyramine, N-trans-feruloyltyramine, and N-trans-coumaroyltyramine for inhibition of α-MSH-mediated tyrosinase activation were 0.8, 20.2, and 6.3 µM, respectively. Overall, N-trans-caffeoyltyramine possessed the strongest anti-melanogenesis activity among the three phenethyl cinnamamides evaluated. The inhibitory effect of N-trans-caffeoyltyramine was verified by determining the melanin content and tyrosinase activity in melanoma after treating the cells with synthetic compounds. Thus, N-trans-caffeoyltyramine isolated from hemp seed extract could be useful in cosmetics as a skin-whitening agent.

Identification of toxicity variations in a stream affected by industrial effluents using Daphnia magna and Ulva pertusa.

A comprehensive toxicity monitoring study from August to October 2011 using Daphnia magna and Ulva pertusa was conducted to identify the cause of toxicity in a stream receiving industrial effluents (IEs) from a textile and leather products manufacturing complex. Acute toxicity toward both species was observed consistently in IE, which influenced toxicity of downstream (DS) water. A toxicity identification evaluation (TIE) confirmed that both Cu and Zn were key toxicants in the IE, and that the calculated toxicity based on Cu and Zn concentrations well simulated the variation in the observed toxicity (r(2)=0.9216 and 0.7256 for D. magna and U. pertusa, respectively). In particular, U. pertusa was sensitive enough to detect acute toxicity in DS and was useful to identify Zn as a key toxicant. Activities of catalase, superoxide dismutase, glutathione peroxidase, glutathione S-transferase, and malondialdehyde were induced significantly in D. magna, although acute toxicity was not observed. In addition, higher levels of antioxidant enzymes were expressed in DS than upstream waters, likely due to the Cu and Zn from IE. Overall, TIE procedures with a battery of bioassays were effective for identifying the cause of lethal and sub-lethal toxicity in effluent and stream water.