Extraction Of Steroidal Alkaloids And Determination Of Main Secondary Metabolites Of Pachysandra Terminalis

Pachysandra terminalis Sieb. Zucc is a species of flowering plant of pachysandra inBuxaceae. As a Chinese folk medicine, it has the effect of dispelling wind and removing cold,promoting blood circulation, and is widely used to relieve the pain and anesthesia of jointscaused by Rheumatism and Bruises because of containing a large number of steroidalalkaloids in which has the anti-tumor activity. The plant of Pachysandra has rich resources,has three species and two variants, and widely distributes in Shaanxi, Guizhou, Gansu, Hubei,Sichuan province, and Japan and the southeast of United States. So it has significant valuethat study the extraction and quantitative analysis of its alkaloids and the system analysis ofsecondary metabolites, that can lay a foundation for the clinical applications of the medicinalherbal and for the further development and exploiting of the plant of Pachysandra.We have established a method for rapid separation and quantitative analysis of steroidalalkaloids, and have studied the difference of contents and distribution of the major secondarymetabolites in this plant. In addition, we have also studied the change of secondarymetabolites in apple branch after pruning with apple tree “Changfu2”and “Yuhua Zaofu” asthe test materials. The main results are summarized as follows:1. We can use the petroleum ether, acetone, and diethylamine solvent (20:5:1and60:5:1)to rapid separate the active ingredients—steroidal alkaloids. Firstly the medicine plantsamples was extracted by ethanol and n-butanol to obtain the total alkaloids, then then-butanol part was fractionated to get the pure single component by dry columnchromatography on silica gel using e petroleum ether, acetone and diethylamine (20:5:1and60:5:1) as the eluent.2. Through using UV spectrophotometric method, we have determinated the contents ofsecondary metabolites which including the steroidal alkaloids in the herbs. The alkaloids ofherbal medicine mainly focus on the underground stems parts, up to22.921mg/g DW, itscontents are nearly twice times than that of other parts of the plant. The average alkaloidcontents of the whole plant were only12.444mg/g DW, and minimum contents in roots(11.255mg/g DW), so its main medicinal part is the underground stem. 3. Similarly, the sugar of the herbs also focus on the underground stems (10.762mg/gDW), the contents were almost twice times than that of the other parts.4. The flavonoid contents were minimal in underground stems, but and concentrate onleaves (23.514mg/g DW);5. Using the same method, we have also determinate the contents of the cortical totalphenols after the pruning of apple branches, it had an obviously regular change—firstlyrapidly increased, then gradually declined, and then slowly recovered to its basic value，therange of cortical total phenols contents were7.749mg/g～12.612mg/g(FW).