| The plants of Eucalyptus globules Labill. belonging to the family of Myrtaceae distributed widely in Guangdong, Yunnan, Sichuan provinces and Guangxi autonomous region of China. The fruit of Eucalyptus globules Labill.(Yi kou zhong ) has long been used as traditional Chinese folk medicine to treat cold, inflammations, contusions and wounds. It was also used as an anti-tumor medicine in some folk remedy.Pre-phase study on the fruit of Eucalyptus globules Labill. has found that components of this kind of fruit has strong activities. Therefore we continued to investigate the plant instructed by the modernization theory of traditional Chinese medicines(TCM) so as to develop a new kind of medicine.Part 1 Studies on chemical componentsIn order to reveal its substance base, compounds were separated by chromatography on silica gel and Sephadex LH-20 columns. Their structures were determined by spectral analysis (UV,IR,MS,H-NMR,13C-NMR,2D-NMR) and phytochemical properties. 9 compounds were isolated and identified as sistosterel (1) Doucosterol (2) stigmasterol (3) Quercetin(4) Kaempferol (5) ursolic acid (6) betulinic acid (7) betulonic acid (8) 1- (1'-ledol) -isopentane-2, 4, 6-trihydroxy-3, 5-benzendihyde(9). Compounds3, 4, 5 were firstly isolated from this plant and compound 9 was a new compound.In previous studies the essential oil from fruit of E.globules Labill were extracted and 17 components were identified. In this paper steam distillation method was adopted and the chemical components of essential oil were analyzed by GC-MS. 31 components were identified : Pinene, phellandrene, terpinene, limonene, 1,8-cineole, terpinene, cymene, careen, isovaleric acid, isopentyl ester, linaloloxide, aristolene, isoledene, terpienol-4, caryophyllene, elemene, pulegone, longifolene, a -terpineol, carane, eucalyptol, acetic acid, geraniol ester, aromadendrene, umbellulol, cadinol, guaiol, globulol, spathulenol, V -eudesmol, a -eudesmol, carvacrol and iludol.The contents and compositions of amino acids in fruit of E.globules Labill were determined by amino acid analyzer. The results showed that there were at least 17 kinds of amino acids including 7 kinds of essential amino acids to human body and the main components were asparagic acid, glutarnic acid, glycosthene, valine, leucine and arginine. The total amount of amino acids was 4.76%(g/g) and the essential amino acids was 2.11%, accounting for 44.3%.The content of free amino acids was 25.29mg in every 100g fruits of E.globules Labill and the free essential amino acids was 9.3mg, accounting for 36.7%.Part 2 Studies on quality standard of ETAThe previous study showed that the fruit of E.globules Labill was generally rich in triterpenoid acids which showed strong pharmacological activities. To meet the higher requirements in developing new medicine guided by state drug administration, the extraction process of Eucalyptus triterpenoid acids (ETA) was optimized according to the orthogonal design, with ursolic acid, betulinic acid and betulonic acid as the quantitative marker. The main influencing factors, i.e., ethanol concentration, ethanol volume, ethanol flow speed and soaking time were systematically optimized. The studies of extraction process had settled a solid base for the industrialization of ETA.In order to control the quality of ETA, the content of ETA was determined by UV spectrophotometry and HPLC methods for the first time, besides identification and test items. The quality standard of extracted ETA was then established, which guaranteed stability, controllability and reproducibility of the ETA sample.Part 3 Studies on pharmacology effects of extractions from fruit of Eucalyptus globules Labill.MTT assay was adopted to examine the inhibitory effects of ETA on the growth of different tumor cell lines in vitro for the first time. The growth inhibitory ratio of ETA on the following tumor cell lines reached above 85%: lung cancer (A549), cervix carcinoma (Hela), melanoma (B16), hepatocarcinoma (HepG2, SMMC-7721,BEL-7402, BEL-7404), colon cancer (L...
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