Separation Of Effective Components In Juniperus Sabina By High-speed Counter-current Chromatography

Juniperus sabina belongs to Cupressaceae Juniperus plants. It can keep water from running away and clean the air in environmental protection, has insecticidal activity in agriculture, and has anti-tumor and anti-viral activity in medicine. Researchers at home and abroad have studied main components in J. sabina and achieved certain results. Terpenes, lignans and flavonoids (including polyphenols) in J. sabina are widespread concerned for their medicinal value.High-speed counter-current chromatography (HSCCC) is an advanced liquid-liquid partition chromatograpy that uses no solid support matrix. Therefore, it eliminates irreversible adsorptive loss of samples onto the solid support matrix used in the conventional chromtographic column and the metamorphic change of samples. It has high resolution, good repeatability, large sample size and has been successfully applied to the analysis and separation of the natural products.This thesis consists of two main parts. The essential oils was extracted by vapor distillation, and analyzed by GC-MS, while the rest components in the fruits of J. sabina (including the lignans and flavonoids) were extracted using solvents with different polarity, and then separated and purified. Methods to separate podophyllotoxin, deoxypodophyllotoxin and flavonoids were built, and products were analyzed by HPLC and1H-NMR. The two-phase solvent system of carbon tetrachloride/ethyl acetate/methanol/water (volume ratio19:6:16:3) was selected to separate podophyllotoxin and deoxypodophyllotoxin simultaneously and the the two-phase solvent system of petroleum ether/ethyl acetate/methanol/water (volume ratio4:4:5:4) was selected to separate flavonoids. The optimal condition is shown as follows. The upper phase was stationary phase and the lower phase was mobile phase. Rotational direction was clockwise and rotational speed was800rpm. The elution direction was from the head to the end. The flow rate was3.0mL/min. The effluent was detected at280nm.Through the study of methods to separate components from the fruits of J. sabina by HSCCC, podophyllotoxin and deoxypodophyllotoxin were separated simultaneously for the first time. The development of this method is value of reference to separation of two or more components in one solvent system, and provides a practical way to prepare high-purity reference standard needed by quality control of herbal medicine.