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摘要：鸦胆子为苦木科植物鸦胆子的果实，主产于广东、广西等地。是一种传统中药，具有清热解毒、截疟、止痢、腐蚀赘疣等功效。我国民间早在古代就广泛应用鸦胆子治疗阿米巴痢疾等疾病。近年来，鸦胆油子作为一种辅助抗肿瘤的药物，引起广泛关注。本文主要对鸦胆子油的提取、分离、鸦胆子油乳样品的试制及其体外的抗肿瘤效果进行了研究．
采用气相色谱法分析了两种不同提取方法得到的鸦胆子油的脂肪酸组成，结果发现提取方法不同对鸦胆子油的脂肪酸组成并无影响。鸦胆子油中的脂肪酸主要包括棕榈酸10.12%，硬脂酸7.39%，油酸60.50%，亚油酸20.08%，另外鸦胆子油中还含有少量的花生酸和蜡酸。采用气相色谱-质谱联用法分析了鸦胆子油的化学组成，共检测出了30种化合物，除脂肪酸外，在有机溶剂提取的鸦胆子油中还检测出一些烃类化合物。
采用常规方法对鸦胆子油精炼，包括脱胶、脱酸、脱水、脱色等工艺。传统溶剂法提取所得到鸦胆子油色泽较深，含有较多的色素，本文考察了脱色剂种类、脱色剂用量、脱色温度、脱色时间对鸦胆子油脱色效果的影响。罗维朋比色法检测结果表明油脂脱色环节对脱色效果影响按大小排序依次为脱色温度，脱色时间，脱色剂种类，脱色剂用量。通过正交实验确定了鸦胆子油脱色的最佳工艺条件为脱色温度100℃，脱色时间30分钟，SF-NO型脱色剂，脱色剂用量1%。
本文还研究了不同制备方法所得鸦胆子油乳样品的稳定性，分别选用大豆磷脂和Tween80两种亲水性不同的乳化剂，考察其对鸦胆子油乳样品乳化效果的影响。结果表明，以大豆磷脂为乳化剂，鸦胆子油：甘油：水为4：1：35时，鸦胆子油乳样品在60℃水浴中能保持稳定1小时，在室温下可保持稳定1个月。
通过体外试管实验考察了鸦胆子油乳样品对HL-60人白血病、Bel-7402人肝癌、BGC-823人胃癌和KB人鼻咽癌肿瘤菌株的抑制效果。并考察了脱脂鸦胆子的极性成分对鸦胆子油乳样品体外抗肿瘤效果的影响。实验结果表明：以市售鸦胆子油乳为对照，所制备的鸦胆子油乳样品与市售鸦胆子油乳对Bel-7402人肝癌、KB人鼻咽癌肿瘤菌株均有抑制效果，且与市售鸦胆子油乳的效果相当。添加了脱脂鸦胆子的水提物、氯仿提取物、乙醇和甲醇提取物的鸦胆子油乳样品对上述2种肿瘤菌株的抑制率都有不同程度的提高。加入脱脂鸦胆子甲醇提取物的鸦胆子油乳样品在抑制Bel-7402人肝癌、KB人鼻咽癌肿瘤菌株的同时，对HL-60人白血病和BGC-823人胃癌肿瘤菌株也有较好的抑制作用。另外，体外试管实验还表明，未碱炼的鸦胆子油乳样品的抗肿瘤效果优于碱炼的鸦胆子油乳样品，由此可推断鸦胆子油中游离脂肪酸为抗肿瘤的主要活性成分。
Abstract
Java brucea is the fruit of the plant Java brucea of Simaroubaceous family, mainly originating in Guangdong and Guangxi provinces. Java Brucea Fruit is a kind of Chinese traditional medicine, which has the function of removing heat and detoxification, antimalarial, curing dysentery, and corroding wart. In the ancient China, its people had been using Java Brucea Fruit to cure ameba diarrhea. Recently, as the oil of Java Brucea Fruit was helpful to deal with tumors, it has got much attention. The thesis focused on the extraction and separation of Java Brucea Fruit oil and the trial-producing of oil emulsion while the effect of anti-tumor effect in vitro was studied.
The fatty acid compositions of Java brucea extracted by those two methods were analyzed by GC. With its results indicating that the two different extraction methods had little influence on the fatty acid composition of Java brucea. The fatty acid compositions of Java brucea oil were: palmitic acid 10.12%, stearic acid 7.39%, oleic acid 60.50% and linolic acid 20.08%, and a small amount of arachidic acid and cerotic acid were detected. GC-MS was adopted to analyze the chemical composition of Java brucea oil. 30 compounds were identified. Besides fatty acids, some advanced hydrocarbons were found in the oil extracted with solvent extraction.
Routine method was adopted to refine Java brucea oil. The processes of deglue, deacidification, dehydration, decolor were included. The Java brucea oil extracted by organic solvent has more pigment. The effects of decolorizer kind, decolorizer amount, decoloring temperature and decoloring time were studied. The results of Lovibond test showed that the decoloring temperature had the biggest influence on the decoloring effect, the second was decoloring time, the third was decolorizer kind and the last was decolorizer amount. A series of parameters optimized by orthogonal experiment were as follows: decoloring temperature, 100℃; decoloring time, 30mins; decolorizer, SF-N0; decolorizer amount, 1%.
The stability of Java brucea oil emulsion prepared by different methods was studied. Soybean lecithin and Tween80 were the emulsifiers. Their influences on the emulsifiability of Java brucea oil emulsion were studied. The results showed that when soybean lecithin was the emulsifier and the proportion of Java brucea oil, glycerine and water was 4:1:35, and the emulsion’s stable period was 1 hour at 60℃, or 1 month at room temperature.
The anti-tumor effects of the emulsion to HL-60 leukosis, Bel-7402 hepatic carcinoma, and BGC-823 gastric cancer and KB nasopharyngeal darcinoma were studied in vitro test. The restraint effect of polar ingredients of fat free Java Brucea on the tumor strain was learnt. The result indicated that in contrast to the Java Brucea oil emulsion sold in the market, the emulsion we prepared could depress the growth of Bel-7402 hepatic carcinoma and KB nasopharyngeal darcinoma strain too, and the effect was equivalent. The ingredients in fat free Java Brucea were extracted by water, ethanol, CHCl3 and methanol. The emulsion appended the above extracts could improve the anti-tumor effect of emulsion to different extent. The Java Brucea oil emulsion appended methyl extracts could restrain Bel-7402 hepatic carcinoma, KB nasopharyngeal darcinoma, and HL-60 leukosis and BGC-823 gastric tumor strain at same time. In addition, the anti-tumor effect of deacidified Java Brucea oil emulsion was less than that of non-deacidification in vitro test. So it could be inferred that the free fatty acids of Java Brucea oil were the main active components |
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发表于 2006-11-25 10:21:26
