Screening Of Medicinal Plants Rich In Rosmarinic Acid And Separation Of Target Components

Rosmarinic acid is a natural antioxidant.In the experiment,the medicinal plants rich in rosmarinic acid were used as raw materials,and the yield of rosmarinic acid was used as the detection index to establish the chromatographic conditions for the determination of rosmarinic acid by HPLC.Optimization of rosmarinic acid and salviaflaside from the spikes of Prunella vulgaris L.by ultrasonic cell grinder extraction and response surface design;enrichment and purification of rosmarinic acid and salviaflaside by aqueous two-phase extraction.The research contents are as follows:1.The HPLC method for the determination of rosmarinic acid was established and the chromatographic conditions were as follows:the wavelength was 325 nm;column temperature at 25?;1.0 mL/min flow rate;10 ?L injection volume;methanol-0.1%trifluoroacetic acid(40:60,v/v)was used as the mobile phase and 30 min elution time.Under this condition,the peak of rosmarinic acid is good.2.Ultrasonic assisted extraction technology was used to extract and determine the content of rosmarinic acid from various medicinal plants.Through the response surface optimization experiment,the best extraction conditions of rosmarinic acid in different medicinal plants are:aerial parts of Lycopus lucidus Tilrcz.var.hirtus Regel(ALL):liquid solid ratio 20 mL/g,ultrasonic irradiation power 250 W and ultrasonic irradiation time 30 min;roots and rhizomes of Salvia miltiorrhiza Bge.(RRSM):liquid solid ratio 20 mL/g,ultrasonic irradiation power 250 W and ultrasonic irradiation time 40 min;the fruit-spikes of Prunella vulgaris L.(FPV):liquid solid ratio 20 mL/g,ultrasonic irradiation power 250 W and ultrasonic irradiation time 50 min;leaves of Perilla frutescens(L.)Britt.(LPF):liquid solid ratio 25 mL/g,ultrasonic irradiation power 250 W and ultrasonic irradiation time 50 min;fruits of Perilla frutescens(L.)Britt.(FPF):liquid solid ratio 25 mL/g,ultrasonic irradiation power 250 W and ultrasonic irradiation time 40 min;the volume fraction of ethanol 50%and the temperature 20?.Under the best extraction conditions,the content of rosmarinic acid were:7.78±0.06 mg/g,1.87±0.04 mg/g,5.79±0.08 mg/g,1.71±0.03 mg/g and 2.68 ± 0.57 mg/g,respectively.3.Rosmarinic acid and salviaflaside were extracted from the fruit-spikes of P.vulgaris L.by ultrasonic cell grinde extraction technology.The effects of different factors on the yield of the target product were investigated by kinetic experiments and response surface experiments.The best extraction technology of rosmarinic acid and salviaflaside from the fruit-spikes of P.vulgaris L.was obtained:50%of the volume fraction of ethanol;25 mL/g of liquid solid ratio;buffer time 2.0 s;cavitation time 1.5 s;ultrasonic irradiation power 310 W;extraction time 6min.Under the optimum conditions,the yield of rosmarinic acid and salviaflaside were 5.66±0.01 mg/g,0.71±0.06 mg/g,respectively4.The purification parameters of rosmarinic acid andsalviaflaside from the fruit-spikes of P.vulgaris L.were studied by aqueous two-phase extraction(ATPE).The extraction ability of four aqueous systems(PEG/(NH4)2 S O4,PEG/Na3C6H5O7·2H2O,C2H5OH/(NH4)2SO4 and C2H5OH/Na3C6H5O7·2H2O)was compared,PEG/Na3C6H5O7·2H2O as the target product in aqueous two-phase system,through the investigation of the mass fraction,temperature and neutral salt of the phase,the best process parameters of aqueous two-phase extraction were determined:The mass fraction of PEG600 is 20%,the mass fraction of Na3C6H5O7·2H2O is 22%,40? of the extraction temperature,the mass of neutral salt NaCl is 0.4g,and 15 min of the homogenate assisted extraction.Under the optimum extraction conditions,the extraction rates of rosmarinic acid and salviaflaside were 92.3%± 0.03%,90.7%± 0.02%,respectively;through aqueous two-phase extraction,the purity of rosmarinic acid and salviaflaside is increased by more than 2 times.In this study,the medicinal plants rich in rosmarinic acid were screened and the chromatographic conditions for the determination of rosmarinic acid by HPLC were established.The extraction process of rosmarinic acid and salviaflaside from the fruit-spikes of P.vulgaris L.was optimized by directly contacting the ultrasonic probe with the plant tissue and the extraction solvent and enrichment and purification of target components by aqueous two-phase extraction.This method is green and low cost,which lays a theoretical foundation for the efficient extraction and separation of rosmarinic acid.