| Huperzine A(Hup A)is an alkaloid extracted from Huperzina serrata Thunb.Trev.It can effectively improve the efficiency of repair of mental activity,memory impairment,has obvious therapeutic effect on age-related memory decline and Alzheimer’s disease,and all obstacles and memory enhancement have improved role in alleviating the slow development of adolescent learning,memory,has high medical value and economic value.For a long time,the study on production technology of huperzine A has been in low level,with the increase in market demand and the depletion of resources,how to make rational use of resources Huperzina serrata Thunb.Trev,maximize extraction of huperzine A,improve the utilization rate has become the research topic.This study investigated on yield and purity of huperzine A,comparison of the extraction methods,selection of alcohol extracted as the optimized extraction technology.Select the ethanol times,ethanol concentration,pH as mainly factors to response surface optimization analysis,get the optimum extraction conditions of huperzine A.At the same time,by the pilot-scale experiments,provide experimental basis and rational suggestion for the industrial production of huperzine A.(1)extraction method and single factor testThe acid water method and ethanol method was comparisoned,pay close attention to huperzine A purity and recovery,process operation difficulty level and energy consumption,the alcohol extraction method is beneficial to the realization of industrialization.Select the ethanol concentration,ethanol times,pH as three factors,through the single factor experiment,preliminary to determine the effects of various factors on the purity and recovery rate.(2)the optimal response surface analysisThrough the single factor experiment,the concentration of ethanol(A),ethanol times(B),pH(C)as the three factors,and to determine the level of each factor,with huperzine A purity(Y)and recovery(Z)as response value,optimized the conditions of extraction process by the response surface design.The main factors of the two coding multiple regression equation:Y=34.58656-0.70323A-2.41725B+2.02862C-9.75*10-3AB-4.125*10-3AC+0.12625BC+5.22250*10-3A2+0.10225B2-0.14319C2,Y is huperzine a purity.Z=183.02406-2.31663A-29.65125B+24.49562C-0.20575AB-0.050500AC+0.79250BC+0.028487A2,+2.15375B2-1.40969C2,Z is huperzine a recovery.Modelsignificantly,by analyzing the response surface optimization,the optimal level of three factors were:ethanol concentration 90%,7 times the amount of ethanol times,pH=9.(3)Pilot-scale experiments in silica gel column chromatography purificationHuperzine A crude product was purified by silica gel column chromatography,optimization of mobile phase and sample volume,results show that,in the mobile phase 5:100=ethanol/cyclohexane,sample quantity of crude 0.4mg/mg silica gel conditions,huperzine A yield and purity were:70.09%and 46.57%.In pilot-scale experiments,the mobile phase condition is 4:100,the sample volume of coarse 0.35mg/mg silica gel,the recovery rate was 71.45%,the purity was 47.43%. |
