| Plumula Nelumubinis, the embryo of the seed of Nelumbo nucifera GAERTN., has many pharmacological activities such as alkaloids, flavonoids, polysaccharides, amino acids and so on. So far, most of the researches focus on the alkaloids. Less study has been done on its flavonoids, polysaccharides and amino acids. To achieve a much better utilization of Plumula Nelumubinis, the study was conducted to isolate and purify flavonoids and polysaccharides, to determine pesticide residues and amino acids. The chief contents are as follows:1. Optimization about the Reflux Extraction Processes of Total Flavonoids in Plumula NelumubinisOn the basis of single-factor test, orthogonal design was carried out to optimize the extraction process in which the total flavonoids content and the yield of extraction were taken as the indexs. According to the results, the optimal extraction process was with an amounts of20times70%ethanol extracted two times and the first time was1.5h, the second time was1h.2. Separation and Purification Technology of Total Flavonoids from Plumula Nelumubinis by Using ResinTen types of resins such as ME-2, OU-2, DM130were evaluated by measuring adsorbance and desorption rate of total flavonoids. DM130macroporous resin was finally chosen. Static adsorption and dynamic adsorption were used to determine the best process condition. The optimization of purification conditions of DM130resin were concluded as follows:sample concentration was0.4g·mL-1,pH was3～4, eluent was70%ethanol, the velocity of eluent was1mL·min-1, the elution volume was triple sample solution. With the optimized process, the content of total flavones was more than42%.3. Determination and Purification of Polysaccharides in Plumula NelumubinisCrude polysaccharides from Plumula Nelumubinis were obtained by hot water extraction, deproteinization, alcohol precipitation and vacuum drying. And they were isolated by DEAE cellulose chromatography-32and Sephadex G-100, distilled water and different levels of NaCl were used as eluent, elution curve and seven polysaccharides mixture were obtained.4. Determination of Amino Acids in Plumula Nelumubinis by Gas Chromatography-Mass SpectrometryA method was developed to determine free amino acids and hydrolytic amino acids in Plumula Nelumubinis by GC-MS. Amino acids with derivatization of silanization were separated and identified on HP-5MS column(30m×0.25μm×0.25mm). The initial column temperature of100℃was held for3min, then programmed at8℃·min-1to300℃and maintained at this temperature for further6min. Identification was performed on the basis of the retention time and the mass peak intensity ratio. Quantitative determination was performed using the internal standard technique and the internal standard was sarcosine. The spiked recoveries of the samples were in the range of85.7%-109.2%and the relative standard deviations were0.77%-4.85%.15kinds of hydrilyzed amino acids and13kinds of free amino acids were found. The total contents were7.14%and0.13%,respectively. The method which could analyze amino acids in Plumula Nelumubinis was simple, accurate and sensitive.5. Determination of Organochlorine Pesticide Residues in Plumula Nelumubinis by Gas Chromatography-Mass SpectrometrySamples were analyzed By GC-MS after purified by C18and sulfonated. Identification was performed on the basis of the retention time and characteristic ions. The chromatographic column used in this method was capillary column of HP-5MS (30m×0.25μm×0.25mm). Temperature programming was used and the split ratio was1:1. The spiked recoveries of the samples were in the range of87.9%-105.3%and the relative standard deviations were below6%. The results showed that there was no organochlorine pesticide residues in five samples from different areas. |
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