Study On The Preparation Process Of Total Saponins In Lysimachia Capillipes Hemsl. And The Establishment Of Quality Standards

Herba Lysimachiae Capillipe was the entire dry herb of Lysimachia capillipes Hemsl., it is used in folk to treat rheumatism, irregular menstruation, asthma and also has the effect of tonic, vermifuge and anti-tumor.In order to better explore the medicinal valve of Lysimachia capillipes as a new anti-tumor herbal drug, we aimed the triterpenoid saponins as target and established a method using HPLC-ELSD to detect the content of saponins, which took Capilliposide B and Capilliposide C as index compounds. By analysis of saponin content, the extraction and purification process were optimized, and the quality standards of herb and intermediate were also developed. The research was aimed to explore new anti-tumor drug with well-defined ingredient and high efficiency.By comparing the traits of herb with different collected location and time, an identification method of medical material was consummated. We set the limitation of saponin content through HPLC analysis of different batches of herb, using the two major saponins as analysis subject. Saponin content in different parts of plant was also tested. The moisture content was determinate by torrefaction method. The minimum of saponin content was 4.00% and the maximum of moisture content was 12.00%.The extraction process was screened by combination of single factor experiments and orthogonal design. The optimal extraction process was as follows:The herb was soaked in 70% ethanol for 30 min before the first extraction, then extracted with 20 times of solvent for 1 h, finally with 16 times for 0.5 h. The yield and extraction rate were 5.31% and 91.2% respectively.HPD450 macroporous resin was selected to be the suitable resin by dynamic adsorption and desorption test. The final purification process was optimized by orthogonal design using the combination of saponin yield and content as composite index. The final purification process was as follows:After concentrating to 0.25g·mL-1 with pH level 7-8, the extract was loaded to macroporous resin column with diameter-height ratio of 1:4, in the speed of 3 BV·h-1. After 3 times of loading repetition and 0.5 h of adsorption, the column was eluted with water,40% ethanol, 70% ethanol for 4 BV respectively in the speed of 4 BV·h-1,4 BV·h-1,6 BV·h-1. The elute of 70% ethanol was dried then yielded intermediate.5 batches of intermediate were prepared by using the selected process. The quality of intermediate can be measured by the total content of Capilliposide B and Capilliposide C. Through the analysis of different batches, content limitation of total saponin was formulated as 55.00%.The flowability and hydroscopicity were measured during the selection of adjuvant. Using the saponin intermediate and microcrystalline cellulose as material, Capillipe Enteric-coated Capsules were prepared. The saponin limitation in capsules was 434.5 mg·g-1. The quality standards of capsules were also established. Accelerated stability index of initial month were investigated.