| In recent years, domestic and overseas scholars are increasing interested in researching natural, safety and health medicine and food plants, Leaves of Morus alba L. comes from the plant of Morus alba L. which sweet-bitter flavor and cold-natured. Morus alba L. has function of dispelling wind and heat, clearing liver and improving vision, clearing lung-heat and moistening dryness. Modern research has shown that mulberry leaves have fall hematic fat, fall blood sugar, anti atherosclerosis, antiviral, anti-inflammatory, anti-aging and other pharmacological effects. Taking mulberry leaves as the research object, the total phenols and total alkaloids of mulberry leaf on extraction and purification. In this paper, optimization of the extraction and purification from mulberry leaves to get optimum parameters of total phenols and total alkaloids by consulting relevant literature and combining with the characteristics of the lab,To develop diabetes drugs has important application value. Based on this, the main contents are as following:1, With four selection of extract solvent, rutin content was very low of alkaline reflux and alkaline diacolation extract, rutin and chlorogenic acid content in alcohol extract was more than two times of the water extraction. Using orthogonal experiment method to screen out the best extraction parameters, the experimental results has shown that:the best condition for 12 times and 10 times with 70% ethanol, decocting 2 times and 2 hours every time.2, Two kinds of cationic resin were compared:metastatic rate of DNJ of Cation resin 732 was higher, but repeated use effect was poorer. The optimum parameters of purification of total alkaloids were screened out by single factor experiment: Used cation resin D001,1.2BV drug and 4BV 2% ammonia was used, swelling time was 9 hours the flow rate was 10 mL/min. Optimal sample amount for the anion resin was 0.67 BV. The DNJ of three results was 25.8±1.5, which showed good stability.3, Based on static adsorption and dynamic adsorption, AB-8 was sifted out from eight types of macroporous resin. The optimum parameters of purification of total phenols were screened out by single factor experiment. The parameters in the first tests:the ratio of crude drug and macropourous resin was 0.8:1 (g/mL), the sample concentration was 250mg/mL, pH was 2.0 and the volumn of 0.5% ammonia water was 3BV. The parameters in the second tests: the ratio of crude drug and macropourous resin was 1.25:1 (g/mL) and the volumn of 50% ethanol water was 3BV. The results of the validation was 52±1.5%, which showed good stability.4, The qualitative analysis of the main chemical compositions of total phenols of mulberry leaves were determined by HPLC-ESI-MS. MS spectrum revealed and UV absorption spectrum that twelve compounds were identified: neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeoylshikimic acid(isomers), rutin, kaempferol-3-O-rutinoside, kaempferol-3-O-glucopyranoside,quercetin-3-O-(6"-O-acetyl)-β-D-glucopyranoside, kaempfer-ol-3-O-(6"-O- acetyl)-β-D-glucopyranoside, quercetin and kaempferol.5, The qualitative analysis of the main chemical compositions of total alkaloids of mulbe-rry leaves were determined by UPLC-ESI-MS. MS spectrum revealed and eference substance compared the main four compounds of total alkaloids were entified preliminary:1-DNJ,2-O-β-Glc-DAB, fagomine, N-Me-1-DNJ.6, Followed the optimized process, we performed the pilot scale experiments three times. 10 kg muberry leaves was used, more than 14%o extraction of total phenols and more than 1.5‰ extraction of total alkaloids was obtained respectively, the grade of purity was more than 50% and 26%(DNJ). the pilot scale experiments has shown the process was stable. |
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