Study On The Preparation Technology Of Phillygenin In Forsythia Suspense Leaves

Forsythia suspensa(F.suspense),the air-dried fruit of Forsythia suspensa(Thunb.)Vahl(Oleaceae),has been used for more than two thousand years in the clinical practice of Chinese traditional medicine.It has the functions of clearing away heat,detoxifying,and dispersing swelling.It is recorded in the contemporary works “Chinese Materia Medica” and “Dictionary of Traditional Chinese Medicine” that "Forsythia suspensa stems and leaves,bitter taste,cold nature,and have the functional indications of heat clearance and detoxification,mainly for cardiopulmonary pyretic accumulation".Studies have shown that the main chemical constituents of Forsy thia suspensa leaves have great similarity with Forsythia suspensa,but the content is different.For example,the contents of phillyrin and phillygenin are higher than fruits.However,Forsythia suspensa leaves,as by-products of Forsythia suspensa,are mostly not effectively utilized except for in folk production of Forsythia tea drinks,resulting in a great waste of resources.Therefore,it is of great significance to rationally and fully exploit the resources of Forsythia suspensa.Modern pharmacological studies have shown that Forsythia suspensa leaves have many activities such as anti-inflammatory and liver protection.As the main active substance,phillygenin is difficult to extract and separate directly which is relatively small in natural medicinal plants.In addition,because the content of phillyrin as the glycoside of phillygenin is relatively high,the hydrolysis of phillyrin to obtain phillygenin has been reported.In order to find a direct method for the separati on of phillygenin from Forsythia suspensa leaves,In this paper,the method of using Forsythia suspensa to ferment the Forsythia suspensa leaves and transforming phillyrin in Forsythia suspensa leaves to form phillygenin was studied.In the study,the high purity of phillygenin monomer was prepared by extraction,separation and purification technology.This paper is expected to provide a basis for the industrialized production and application of phillygenin.The details are as follows:1.The Forsythia suspensa leaves in different months from April to October were used as the research objects.The contents and changes of the two main lignans such as phillyrin and phillygenin were detected before and after fermentation by Forsythia suspense leaves.The results showed that phillyrin,the main component of Forsythia suspense leaves after fermentation,could be transformed into phillygenin.The content of phillyrin in Forsythia suspensa leaves was up to 81.29mg/g in May,and the conversion rate of phillyrin and the conversion yield of phillygenin were higher.The content of phillygenin was increased from 4.88mg/g to 41.38mg/g.This study indicates that the use of Forsythia suspensa leaves self-enzymatic fermentation is an effective means of converting phillyrin into phillygenin.2.The fermentation conditions of forsythin in fresh forsythia suspensa leaves were studied by orthogonal experiment.The most appropriated way was: 55% water,fermentation temperature 30?,fermentation time 2 h,forsythia Glycosides are efficiently transfered to phillygenin.The three-time magnification repeat experiment showed that the average content of phillygenin was 41.60 mg/g.Because the fresh Forsythia suspensa leaves are difficult to store,the process is more fitted for newly harvested leaves in the season.3.There was response surface methodology,which was used to research the fermentation process of phillygenin in dried Forsythia suspensa leaves transformed into phillygenin,the most satisfactory parameters were: 50% water,fermentation te mperature 30?,fermentation time 100 min,the lipoprotein was increased by about28 times;the amplifying experiment showed that phillygenin was 46.40 mg/g on average,which was close to the theoretical value under optimal conditions.4.The process of extracting phillygenin by ethanol reflux was explored and optimized.The optimal extraction conditions were as follows: ethanol concentration 80%,solid-liquid ratio 1:15,extraction time 30 min,extraction times 2 times.Amplification and repeated experiments showed that the average extraction yield of forsythin was 3.27%,and the process was reproducible.In addition,the extract of the Forsythia suspensa leaves fermented product was initially purified,and the content of phillygenin was increased 10.39%-31.65% after washing 3 times with 5 times;then washed twice with 5 times 20% ethanol,the content of phillygenin can be increased from 31.65%-39.75%.It is obvious that the initial purification effect is greatly beneficial.5.The above fermentation product was further purified by using a macroporous adsorption resin.Firstly,the adsorption and desorptive effects were used as indicators to screen the macroporous adsorption resin,and HP-20 resin was determined as the separation material.The adsorption and desorption conditions were determined by static adsorption and static desorption experiments: adsorption equilibrium time 2 h,30-min parsing time,and the most suitable temperature was 25 ?.It was found that 70% ethanol eluent could obtain more phillygenin with a gradient of ethanol concentration of different concentrations,and the content was up to 90%,and the yield was 87.27%.Finally,we investigated recrystallized solvent.The results showed that methanol was used as the recrystallized solvent,the purity of phillygenin was 98% after three recrystallized operations.Preliminary results indicate that HP-20 macroporous adsorption is well for separation and purification of phillygenin,and high purity by recrystallizing of methanol.