Study On Preparation Of Ginkgo Biloba Extract Based On Continuous Resin Column Chromatography

Traditional chromatographic separation technology is usually accompanied by low resin utilization,large amount of mobile phase and long production cycle.In this study,Ginkgo biloba Leaves were used as a raw material to establish a complete set of continuous chromatographic device based on the saturated adsorption method to prepare and separate Ginkgo biloba extract(GBE)which met the requirements of Pharmacopoeia.Its main content was divided into the following five parts:1.Through the static and dynamic adsorption and desorption characteristics of six types of resins,such as D101,ADS-17,X-5,AB-8,DM-130 and S-8,the ADS-17 macroporous resin has a higher flavonoid content to Ginkgo biloba Adsorption and desorption effects.The process of preparation and separation of GBE from ADS-17 resin was studied with the index of adsorption and desorption rate of Ginkgo biloba flavonoids.The sample concentration,desorption ethanol volume fraction and pH were 3.15 mg/mL,70% and 5.0,respectively.The volume of sample,wash,desorption were 5 BV,3 BV and 3 BV,respectively.The speed of sample,wash,desorption were 2 BV/h.Under this condition,the contents of total flavonoids and lactones in GBE increased from 2.71% and 0.72% before preparation and separation to30.28% and 6.83% respectively,and the recoveries of total flavonoids and lactones were82.30% and 78.32%,respectively.2.Due to the problems such as low resin utilization and large amount of mobile phase in the preparation of GBE by traditional process macroporous resin,this paper proposes a saturated adsorption method for the saturation adsorption of ADS-17 macroporous resin.When the resin is saturated adsorption,ginkgo total flavonoids and lactones in crude drug amounts of 2.72 to 3.28 times weight per g resin can be adsorbed.Comparing with traditional adsorption resin method,resin usage was decreased by 2/3.After saturated adsorption,the contents of total flavonoids and lactones of GBE were in accordance with the national pharmacopoeia standards.3.Based on the results obtained from single-column resin saturated adsorptionexperiments,the laboratory developed a continuous chromatography device.The core is to cut the original single resin column into three resin columns,which are fixed in series on the disc.The continuous chromatographic equipment consists of twelve reservoirs,six fluid pumps,eighteen resin columns,a logic control valve and a software control system.Eighteen columns are divided into six zones.Six fluid pumps are used to transport various types of liquids.Regular rotation of valve achieves continuous separation of GBE by soft control system.4.The process parameters of preparation and separation of GBE by continuous chromatography principle prototype(Φ5.1 cm×30.0 cm)as follow: A-loading speed 45mL/min,B-loading speed 45 mL/min,C-loading speed 45 mL/min,washing speed 36mL/min,desorption speed 36 mL/min,regeneration speed 18 mL/min,balancing speed 18mL/min,logic control valve switching time 20 min.Under this condition,the content of total flavonoids and lactones in obtained GBE were 24.06~26.01% and 6.06~7.01%,and the equivalent of Ginkgo biloba leaves adsorbed by the resin were 2.58~3.11 kg/kg.5.The process parameters of preparation and separation of GBE by continuous chromatography production equipment(Φ80.0 cm×2.0m)as follow: A-loading speed 1500 L/h,B-loading speed 1500 L/h,C-loading speed 1500 L/h,washing speed 1200 L/h,desorption speed 1260 L/h,regeneration speed 600 L/h,balancing speed 600 L/h,logic control valve switching time 1 h.Under this condition,the content of total flavonoids and lactones in obtained GBE were 24.33~26.01% and 6.18~7.01%,and the equivalent of Ginkgo biloba leaves adsorbed by the resin were 2.87~3.13 kg/kg.The continuous chromatographic device based on independent laboratory development can not only be used for preparing and separating GBE but also for preparing and separating the active ingredients of other traditional Chinese medicines and has broad application prospects in the preparation and separation of the active ingredients of traditional Chinese medicines and provides the new ideas of the preparation and separation of the active ingredients of the traditional Chinese medicines.