The Wormicidal Sibstance of Fresh Water Fishes on Clonorchis sinensis(VI) - Struture Identification of Purified Clonorchicidal Sudstance from Epidermal Mucus of Cyprinus carpio nudus

간흡충에 대한 살충성물질에 관한 연구 (VI) - 향어 체표면 점액내 살충성물질의 구조 확인

In order to elucidate the defence mechanism of Cyprinus carpio nudus to Clonorchis sinensis, clonorchicidal substance in the epidermal mucus of this fish was isolated by silica gel column and thin layer chromatography and analyzed for its chemical nature by UV, IR and NMR-spectroscopy. The epidermal mucus of the fish was extracted by ethyl ether and separated into 2 fractions by column chromatography using petroleum ether/chloroform(30/70,v/v) as first solvent and methanol as second solvent. The second fraction with greyish white colour showed clonorchicidal effect. The second was again fractionated into greyish white precipitate and clonorchicidal greenish yellow supernatant fraction, by adding petroleum ether and standing the mixture for 5 days at $5^{\circ}C$. The latter was divided into 7 fractions in column chromatography with acetone/ benzene (10/90, v/v) as carrier. The fraction on equivalent to the spot of Rf. 0.225 value among them disclosed the strongest clonorchicidal effect. By this purification procedure, clonorchicidal substance was purified 154-fold with 0.9% yield from the mucus of the fish, and 10mg of the final fraction killed tne metaceicariae in 22 min. Infra red, nuclear magnetic resonance and ultraviolet spectrometric analysis of the purified substance revealed that the substance is linoleic acid. According to the results of the present studies it seemed that this fish could not be proper intermediate host of Clonorchis sixensis, and that defence mechanism of this fish to the worm seems to be correlated with clonorchicidal substance in epidermal mucus.