| Macleaya cordata(Willd.)R. Br.(Macleaya cordata)is a medicinal plant in the family of papaveraceae,which has detumescence,detoxification,and insecticidal effects.The main active compounds in Macleaya cordata are alkaloids,which have been found to possess biological and pharmacological activities.Therefore separation and purification of alkaloids in Macleaya cordata is of great significance for fully studying the biological activity of alkaloids,and it can also achieve comprehensive utilization of this plant.Because both stationary phase and mobile phase are liquid,high speed counter-current chromatography(HSCCC)has obvious advantages including no irreversible adsorption,large injection volume,and a wide range of solvent systems,which has been widely used in separating active compounds from natural product.Ionic liquids(ILs)have gained extensive attention due to their distinct features,such as high thermal stability,tunable viscosity,moderate dissolvability in organic compounds,and could form biphasic liquid systems with numerous solvents.In this paper,using ionic liquid as a modifier of the solvent system,a method for the separation of alkaloids from the fruits of Macleaya cordata by HSCCC was established.Meanwhile,as the characteristic polyphenol in apples,phlorizin has variety biological activities.The HPLC method for the determination of phlorizin in apple products like apple crisp and apple jam was established,which could meet the need of evaluation and quality control of these apple products.In addition,this method was applied for recommended national standards.This article has carried out research on these two aspects,the main contents are as follows:1.The extraction method and HPLC analysis method of alkaloids in the fruits of Macleaya cordata were established,and the separation results of HSCCC and ILs modified HSCCC on alkaloids in the fruits of Macleaya cordata were compared.The partition coefficient of five target compounds in the solvent system dichloromethane-methanol-hydrochloric acid aqueous solution in different volume ratio were investigated,and the basic solvent system was dichloromethane-methanol-0.3 mol/L hydrochloric acid aqueous solution(4:2:2,v/v),then the injection amount of crude extract was optimized.The effect of the type and amount of ILs on the separation effect of five target alkaloids was investigated.Finally,a ionic-liquid-modified HSCCC method was successfully established,using the solvent system dichloromethane-methanol-0.3 mol/L hydrochloric acid aqueous solution-1-butyl-3-methylimidazole tetrafluoroborate(4:2:2:0.015,v/v),five alkaloids including protopine,allocryptopine,sanguinarine,8-O-demethylchelerythrine,and chelerythrine with the purities of 96.33%,95.56%,97.94%,96.22%and 97.90%were separated from the crude extract of the fruits of Macleaya cordata,respectively.2.The HPLC method for determination of phlorizin in apple products like apple crisp and apple jam was established,and applied for recommended national standards.The extraction method of phlorizin in apple crisp and apple jam and the standard curve of phlorizin reference materials were established.Then the precision,stability,repeatability,and the recovery of spiked samples were tested.The phlorizin had a good linear relationship in the concentration range of 5?500 mg/L,and the correlation coefficient(r2)was greater than 0.999.The sampling amount of apple crisp was 50 g,the determination limit was 1.37 mg/L,the limit of quantification was 4.57 mg/L,and the content of phlorizin was 9.50 mg/kg.According to sample measurement method,the recovery rate was 93.79%?99.93%,and the relative standard deviation was less than 5%.The sampling amount of apple jam was 100 g,the determination limit was 0.69 ?g/mL,the limit of quantification was 2.28 mg/L,and the content of phlorizin was 1.18 mg/kg.According to sample measurement method,the recovery rate was between 90.56%?98.78%,and the relative standard deviation was less than 5%. |
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