| Plumula Nelumubinis, the embryo of the seed of Nelumbo nucifera GAERTN., has been extensively used as a traditional medicinal herb in China for thousands of years. It has many pharmacological activities such as the alkaloids, the flavonoids, polysaccharides, volatile oil and so on, which was primarily used for nervous disorders, insomnia, high fevers with restlessness, reversing multidrug resistance and cardiovascular diseases such as hypertension and arrhythmia. The previous research were mainly focused on extraction, isolation, purification and pharmacology activity of alkaloids, the research on flavonoids were somehow ignored. More important, the organic solvent to separate the alkaloids are usually poisonous, and the procedures are cumbersome. To make full use of the medicinal and overcome the weakness of the existing technology, this project try to establish a new method to separate alkaloids, and to enrich the flavonoids at the same time. It is quick, simple, environment-friendly and with high utilization of medicine. The chief contents and results of this research are as follows:1. Precipitation of Plumula Nelumubinis Extract with ZTC1+1-ⅡClarifiersIn this part, ZTC1+1-Ⅱnatural clarify agents was used to remove impurities such as chlorophyll, mucilage glue, proteins and tannins from Plumula Nelumubinis. On the basis of single-factor test, orthogonal design was carried out to optimize the precipitation process in which the solids removal rate and remaining rates of neferine as indexs. According to the results, the optimal process was condensing the extract solution to 1:6 (herb:herb solution), the adding order of 2 components of ZTC1+1-Ⅱnatural clarifier was to add component B firstly and then add component A, the dosages of ZTC1+1-Ⅱwere 0.8mL·g-1 for clarifier B and 0.4mL·-1 for clarifier A, the reaction temperature of clarifier was 80℃, then stirring 10min and holding in water bath at 80℃for 30min respectively. The solids removal rate was 22.81% and the remaining rates of neferine was 89.42%. We can draw a conclusion that ZTC 1+1-Ⅱnatural clarify agents can be used to effectively remove the impurities of extract of Plumula Nelumubinis.2. Precipitation of Plumula Nelumubinis Extract with chitosanChitosan was used to purify the extract of Plumula Nelumubinis. Orthogonal design and response surface method were comparaed to optimize the precipitation process on the basis of single-factor test in which removal rate and remaining rates of neferine were chosen as indexs. According to the results, the best experimental conditions were as follow: the concentration of extract solution was 1:5 or 1:5.5 (herb:extract solution), the dosages of chitosan were 1.2mL·g-1, the reaction temperature of clarifier was 60℃and the pH value was 6.2. There is no significant differences between the results of optimization.3. Separation and enrichment of alkaloids and flavonoids simultaneously by using ion-exchange resinStatic adsorption and dynamic adsorption were used to screen out the best resin in 4 different types of ion exchange resins,4# resin was finally chosen. Based on the reseach of initial solution volume, the velocity of initial solution and eluent, we optimize the technology of separation and enrichment conditions. With the optimized process, the purity of flavonoids and alkaloids were significantly enhanced to 27.08 and 9.48 times than ever respectively.4. The purification of the flavonoids of Plumula NelumubinisThe separated flavonoids were desorbed by gradient elution 20%, 30%,40%, and 50% ethanol in the polyamide column gradually. The flavonoids components of fraction of 40% ethanol eluent were further separated. Five fractions were collected after the flavonoids are further separated by Prep-HPLC.
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