Extraction,Separation And Purification Of Camptothecin And 10-hydroxycamptothecin From Camptotheca Acuminate Leaves

China has a vast territory,rich resources and a treasure house of natural plants.As a special species,camptotheca chinensis only grows and distributes in China.China also takes it as one of the first key protected species to be protected by law.Camptotheca acuminata is rich in dozens of substances with unique biological activities,among which alkaloids are particularly important.Dozens of alkaloids have been found in the fruit and bark of camptotheca,among which camptothecin and 10-hydroxycamptothecin are represented.camptothecin and 10-hydroxycamptothecin are also found in the camptotheca acuminate leaves.The conventional extraction solvents of camptothecin and 10-hydroxycamptothecin include methanol,ethanol,chloroform and other solvents,which have many disadvantages such as large amount of solvent,high extraction cost,unfriendliness to the environment,and potential safety risks.At the same time,the conventional extraction methods include reflux extraction method,cold immersion method,ultrasonic extraction method,etc.,which have the disadvantages of complex extraction process,long extraction cycle and large energy consumption.Therefore,it is particularly important to find a safe,efficient,environmentally friendly and easy to operate process.Based on the above reasons,camptotheca acuminate leaves,which are rich in resources and can be sustainably regenerated,were selected as the extraction raw material,and the surfactant sodium lauryl sulfate aqueous solution was used as the extraction solvent.The active camptothecin and 10-hydroxycamptothecin in camptotheca acuminate leaves were extracted by combining the ultrasound-microwave assisted extraction technology.At the same time,the 10-hydroxycamptothecin was separated and purified.The main contents and results of the study are as follows:1.HPLC conditions for camptothecin and 10-hydroxycamptothecin were established:the detection wavelength was confirmed by full-wavelength scanning at 265 nm.The chromatographic column was Diamonsil C18 inverse phase column(5 m,4.6 mm×150 mm).The mobile phase was methanol:water=55:45(V/V),the column temperature was set at 25?,the mobile phase flow rate was 1.0 mL/min,and the single injection volume was 10 L.camptothecin and 10-hydroxylcamptothecin have good linear relationship in the measured range.2.Select different kinds of surfactant aqueous solutions:Anionic surfactant,cationic surfactant aqueous solution,zwitterionic surfactant,nonionic surfactant aqueous solution,and methanol,ethanol,chloroform,ethyl acetate in camptothecin and 10-hydroxycamptothecin solubility test,considering cost,is in line with the green chemistry,the late can purify finally chose the surfactant sodium dodecyl sulfate solution as extraction solvent,sodium dodecyl sulfate aqueous solution of camptothecin and 10-hydroxycamptothecin solubility were 25.8?g/mL,79.3 ?g/mL.3.Select camptotheca acuminate leaves to the surfactant sodium dodecyl sulfate solution as extraction solvent,and through the response surface optimization design based on single factor,respectively to investigate microwave time,and solid-liquid ratio,microwave power,the concentration of surfactant,four factors of camptothecin in camptotheca acuminate leaves and the extraction of10-hydroxycamptothecin,through data analysis and processing of camptothecin and 10-hydroxycamptothecin total extraction rate,the optimal process parameters:10 min,microwave time and solid-liquid ratio 1:100,microwave power 700 W,the concentration of surfactant 8 g/L,under the condition of the total extraction yield reached 0.0549%.4.The extract was prepared and the extract was initially purified by macroporous resin.AB-8 macroporous resin was used for elution at the flow rate of 2 BV/h and the equilibrium adsorption time of 7 h without changing pH value at room temperature.The flow rate of 1 BV/h of methanol solution was used for elution,and the amount of elution was 8 BV.5.Using 10-hydroxycamptothecin crude products as raw materials for the purification,is obtained by single factor screening of 10-hydroxycamptothecin content have an impact on three factors,by using the response surface software design experiments,respectively to investigate 10-hydroxycamptothecin rough concentration,deposition time,stirring speed of three factors on the influence of10-hydroxycamptothecin content,get the final optimum technological parameters of purification 10-hydroxycamptothecin:raw10-hydroxycamptothecin 35 mg/mL concentration,deposition time 6 min,stirring speed 2000 r/min,under the condition of 10-hydroxycamptothecin content 85.49%.