Study On Quality Of Brucea Javanica And Pharmaceutical Brucea Javanica Oil

There are different and abundant chemical components in Brucea javacina, one of theTraditional Chinese medicine. They had been report that they show the strongpharmacological activities of anti-leukemia, anti-malarial, Antiphytoviral, antitrypano-somal, anti-tumor, anti-cancer such as the simaroubolides, glycosides, alkaloids, terpenoids,fatty acids, flavonoids, phenylpropanoids and so on. At present, a lot of meaningfullyfundamental researches have been done and more researches will continue to be done. Onlyin clinical the intravenous emulsion and oral emulsion containing Brucea javacina oil(BJO)has been widely used in the treatment of tumors, cancers, gastrelcoma, hydrothorax, ascitesand other diseases. The chemical constituents of BJO was not very clear, and a little studieshad been done on the stability of chemical constituents in the production process includingBrucea javacina fruit, BJO and the preparation of BJO. Only the oleic acid content wasrequired in the quality standard of BJO which could not ensure the quality and efficacy ofthe BJO preparation.Hereby, the constituents and mainly contents had been researched in Brucea javacinafruits (BJF) and BJO. Firstly, the BJF was identified by TLC and microscopy method.Secondly, the gas chromatography method for the determination of BJO was developed.Gas chromatography of FAMEs was performed on a0.32mm(i.d.)×30m fused silicacolumn lined with a0.50μm film of polyethylene glycol (CP-WAX52CB). Samples (1μL)were injected in split mode (split/column flow ratio5:1). The flow rate of carrier gas(nitrogen) was1.0mL/min. The oven temperature was205℃; the injection temperature was250℃; the detector temperature was300℃. Before the fatty acid components of BJO canbe analyzed by GC, it is necessary to convert them to low molecular weight non-polar derivatives, such as methyl esters. The optimum process of methyl esterification of BJOwas established by single factor method and Response surface methodology(RSM): theBJO(3mg) was refluxed with a1M solution of potassium hydroxide in methanol(2mL) attemperature of60℃for25min; the solution was cooled,14%boron trifluoride-methanolsolution was added and the mixture was refluxed at temperature of60℃for2min; thesolution was cooled, the mixture was extracted thoroughly with hexane(2mL) andsaturated sodium chloride solution (1mL) was added and the supernatant was what weneeded. Finally, the methyl esters of BJO were determinate by GC. And the result was: onewas that the oleic acid in BJO produced from Beihai Guangxi(⑧) and Guangxi(⑩) wasless than eight percent, and the oleic acid in other origins were greater than eight percent;another was that the linoleic acid in BJO from Beihai Guangxi(⑧), Hainan(⑨),Guangxi(⑩), Haikou Hainan() was less than three percent, and the linoleic acid in otherproducing area were greater than three percent; the content ratio of oleic acid and linoleicacid were more than2.5.Secondly, the fingerprint of BJO was established by gas chromatography. Gaschromatography of FAMEs was performed on a0.32mm(i.d.)×30m fused silica columnlined with a0.50μm film of polyethylene glycol (CP-WAX52CB). Samples (1μL) wereinjected in split mode (split/column flow ratio5:1). The flow rate of carrier gas (nitrogen)was1.0mL/min. The oven temperature was205℃; the injection temperature was250℃;the detector temperature was300℃. The similarity of fingerprints was evaluated by thefingerprint similarity calculation software based on median. The results demonstrated thatdifferent origins of BJF had a good similarity (＞0.99)，and different batches of BJO had abetter similarity(=1.00). However, only in combing characteristic components andfingerprint similarity results, the good quality control of BJF and medicinal BJO willbecome reality. Otherwise, the hierarchical cluster analysis was used to find the bestpartition of all the BJF and medicinal BJO, and learn about the intrinstic relationship andstatistical laws in BJF and BJO.Otherwise, the method based on gas chromatography of the methyl esters of BJO, withdetection by mass spectrometry was used to identified the11components; To characterize the major ingredients of BJO,42major chromatographic peaks were characterized by theirESI-MS spectra.Lastly, according to research results, the quality standard draft of BJO has beenproposed.