Preliminary Study On Preparation Method And Antidepressant Activity Of Total Coumaroylspermidines In Safflower Dregs

The Chinese medicine safflower is a dry tubular flower of compositae plant Carthamus tinctorius L..It has the pharmacological effects of promoting blood circulation,dredging meridians,anti-inflammatory,analgesic and anti-oxidation.It is the main raw material of Chinese medicine injection which including safflower injection and safflower oral liquid.The water extraction process is mainly used in the process of producing safflower injection or oral liquid.It extracts most of the water-soluble components such as safflower yellow pigment and flavonoid glycosides,which have certain solubility in water.The extracted safflower dregs also contain lipid-soluble chemical components such as red pigments and coumaroylspermidines.Previous studies of our group have shown that coumaroylspermidines of safflower have the effect of inhibiting 5-HT reuptake,which suggesting a potential antidepressant effect.In order to rationally develop and utilize safflower dregs resources and further explore the potential biological activity of coumaroylspermidines,this paper studied the extract method,the preparation method of the total coumaroylspermidines in safflower dregs and the separation of coumaroylspermidines.And the antidepressant activity of total coumaroylspermidines was evaluated by behavioral pharmacology initially.The research contents of this paper are as follows:1.Study on extraction method of total coumaroylspermines from safflower dregs.Using the established high performance liquid chromatography?HPLC?method for quantitative determination of coumaroylspermidines in safflower,taking the total extraction rate of four coumaroylspermidines in safflower dregs as assessment index,the effects of natural lighting,extraction solvent,extraction method,solid-liquid ratio,extraction time and times on the extraction rate of total coumaroylspermidines in safflower dregs were investigated.According to the results of single factor test,the optimal extraction method of total coumaroylspermidines in safflower dregs was determined as follows: 35 times the amount of anhydrous methanol was heat reflux three times for 2 hours,the content of total coumaroylspermidines in safflower dregs was?2.894 ± 0.011?mg/g,which was 36.38% higher than that of the ultrasonic extraction method established previously of coumaroylspermidines in safflower.2.Study on the preparation technology of total coumaroylspermidines from safflower dregs and the separation of coumaroylspermidines.The extraction solvent of total coumaroylspermidines used in the preparation process was further optimized based on the established extraction method of total coumaroylspermidines in safflower dregs.And using the established high performance liquid chromatography?HPLC?method for quantitative determination of coumaroylspermidines in safflower,taking the total content of four coumaroylspermidines in safflower dregs as assessment index,the pretreatment conditions?concentration and washing conditions?and column separation materials for the separation and preparation of total coumaroylspermidines were investigated.On this basis,the total coumaroylspermidines were rapidly purified by medium pressure flash chromatography,and the effects of sample quantity and concentration on the separation of total coumaroylspermidines were investigated.The results showed that on the basis of pretreatment in safflower dregs extract,the optimum conditions for separation and preparation of total coumaroylspermidines by C18 medium pressure flash chromatography were as follows: when elution speed was fixed at 15 m L/min,W1/W2 was 4.65?mg/g?,?W1 is the mass of sample to be separated,W2 is the mass of the C18 column separation material?,the concentration of sample was 90 mg/m L,and the purity of total coumaroylspermidines in the final product was over 50%.In addition,N¹,N⁵,N¹⁰-?E?-tri-p-coumaroylspermidine was isolated during the separation and purification of total coumaroylspermidines.3.Preliminary study on antidepressant activity of total coumaroylsper-midines in safflower dregs.The antidepressant activity of total coumaroylspermidines in safflower dregs was preliminarily evaluated and the effective dose was screened on depression in mice by using the immobility time in tail suspension test?TST?and forced swimming test?FST?of mice as the evaluation index.The results showed that compared with the blank control group,the tail suspension and forced swimming immobility of mice were significantly improved when the total coumaroylspermidines dose was 25 mg/kg and 50 mg/kg.Therefore,it is speculated that total coumaroylspermidines may have antidepressant activity.The results again demonstrated that coumaroylspermidines could improve the behavioral despair of mice.It is concluded that the total coumaroylspermidines of safflower dregs have certain antidepressant activity.