薄层色谱和高效液相色谱联用测定暗纹东方鲀肌肉中肌肽和谷胱甘肽的含量

通过薄层色谱(TLC)与高效液相色谱(HPLC)联用技术鉴定了暗纹东方鲀(Takifugu obscurus)肌肉中存在肌肽和谷胱甘肽,同时利用高效液相色谱法测定了肌肽和谷胱甘肽(GSH)的含量。薄层层析采用的展开剂为正丁醇:乙酸:水(4∶2∶1),层析板为硅胶板。高效液相色谱利用Kromasil C18反相柱分析,流动相为10%的纯乙腈和90%含有0.05%三氟乙酸的超纯水。结果表明:通过薄层色谱和高效液相色谱鉴定了暗纹东方鲀肌肉中肌肽和谷胱甘肽,其中暗纹东方鲀肌肉中肌肽含量约213μg/g(鲜重),还原性谷胱甘肽含量约211μg/g(鲜重)。本法样品无需衍生,操作简便,适合于暗纹东方鲀肌肉中肌肽和谷胱甘肽的测定。本文为生物体内肌肽和谷胱甘肽的研究提供借鉴意义。

Determination of the Carnosine and Glutathione from Takifugu obscurus by Thin Layer Chromatography and High Performance Liquid Chromatography

The thin layer chromatography(TLC) combined with high peRformance liquid chromatography(HPLC) method was established for identification the carnosine and glutathione from puffer fish(Takifugu obscurus) muscle.And the contents of carnosine and glutathione were determination by the HPLC.The TLC used the silicon sheet with the developing solvents n-butanol,acetic acid and water(4∶ 2∶ 1).The samples were separated on Kromail C18 column at 30 ℃.An isocratic elution of 90% acetonitrile and 10% Mili-Q water containing 0.05% trifluoroacetic acid was used.The results showed that the carnosine and glutathione(GSH) existed in Takifugu obscurus muscle.The content of carnosine and glutathione(GSH) in Takifugu obscurus muscle was 213 μg/g(fresh weight) and 211 μg/g(fresh weight),respectively.This method was very simple,convenient and operated without derivant.It was suitable for the determination of carnosine and GSH.This study has important reference significance to research the carnosine and glutathione from other organisms.