Studies On Extraction, Separation, Purification And Anti-tumor Activity Of Effective Parts In Brucea Javanica

Brucea Javanica fruit is the seed of Brucea Javanica Merr (L.), which belongs to Brucea species of Simaroubaceae department. In recent years, as a natural anti-tumor drug, Bruea Javanica has got great attention, which is paid by the researchers. This paper mainly focus on extraction, separation, purification and anti-tumor activity of effective parts in Brucea Javanica. And the main results are as follows:Using the method of Solvent reflux extraction, with anhydrous ethanol as the extraction solvent, the extracting technology of water-soluble polar constituents of Brucea Javanica was studied. And the extracting technology was optimized by L9(34) orthogonal experiment with the extracting rate of bitter extracts as index, and with the extracting temperature, ratio of liquid and solid, extracting time and extracting times as factors. The optimal conditions for extracting technology were as follows: extracting 3 times, adding seven-fold amount of anhydrous ethanol,2h each time and extracting temperature 60℃. In that conditions, the extraction rate was reached to 17.4%.A method for the determination of total triterpenes from Brucea Javanica was established by vanillin-glacial acetic acid and perchloric acid colorimetric spectrophotometry. From the instrument precision, repeatability, stability and average recovery, the method feasibility was studied, and the main results are as follows:RSD of instruments precision 0.70%, repeatability 1.32%, stability 4.19%, average recovery 1.17%, and the four of RSD are less than 5%.It indicates that the measured data is desirable, the results are good reproducibility and it provides a method to establish the internal quality control for Brucea and its extracts.And the average content of total triterpenes of bitter extracts, which is from crude anhydrous ethanol extracts of Brucea Javanica, is 3.68%.Through the method of liquid-liquid partition extraction, the bitter extracts of Brucea Javanica was divided into six parts, Cp1, Cp2, Cp3, Cp4, Cp5 and Cp6; and then through the test of inhibition of in vitro cell proliferation to find that the part of chloroform layer Cp2 was the strongest. Then the Cp2 part was purified by silica gel column, and the part was divided into four components, Cp2a, Cp2b, Cp2c and Cp2d. And with the four cell lines of HL-60 human leukemia, BGC-823 human gastric cancer, BeL-7402 human hepatocellular carcinoma, KB human nasopharyngeal carcinoma, testing the anti-tumor activity of the above four components, measured by inhibitory rate of cell proliferation, to determine which components were most active; and the results were showed that:in the same concentration, in terms of four kinds of cell lines, Cp2d is stronger than Cp2c in the cell proliferation inhibitory effect, while Cp2c is stronger than Cp2a and Cp2b.With Brusatol as the reference substance, using the HPLC instrument to analyze the composition of Cp2d, and the results are showed that:the main constituents of Cp2d are Brusatol.