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冷榨文冠果种仁油的毒理学评价及体内外抗氧化功能的研究

Studies on the Toxicological Evaluation and in Vtiro and Vivo Antioxidation Function of Xanthoceras Sorbifolia Bunge Kernel Cold Pressing Oil

【作者】 范雪层

【导师】 邓红;

【作者基本信息】 陕西师范大学 , 食品科学, 2009, 硕士

【摘要】 文冠果(Xanthoceras sorbifolia Bunge)又名木瓜、文登阁、僧灯毛道,为无患子科文冠果属的落叶乔木或灌木,原产中国北方。文冠果具有较强的适应性和抗逆(抗寒、抗旱和抗盐碱)能力,是中国特有的珍稀木本油料及能源、观赏植物。文冠果种子含油率30.4%,种仁含油率达66.39%。文冠果种仁油中不饱和脂肪酸含量高达94%,因而文冠果油具有良好的软化血管及防治心血管系统疾病的作用和具有更好的降胆固醇的作用,可作为天然不饱和因子、不饱和添加剂使用。文冠果油在民间已有较长的食用历史,但其食用安全性的理论研究尚为空白,对文冠果油功能特性的研究也很少。本论文用冷榨法制取文冠果种仁油,对此油脂进行毒理学评价(急性毒性试验、遗传毒性试验、30天喂养试验),从而确定文冠果种仁油的食用安全性,在此基础上进一步对文冠果种仁油的功能特性进行研究,为开发新的功能性食用油资源提供实验依据。试验结果表明:1、在压力55 MPa~60 MPa,仁壳比为9:1,压榨时间为8h的条件下,文冠果冷榨油的得率为40.44%;文冠果油的折光指数为1.4723,相对密度为0.914,与其它植物油相比较低;酸价和过氧化值分别为0.601 mgKOH/g和0.16 meq/Kg,说明文冠果冷榨油新鲜无质变;碘价高达121.4g/100 g,说明文冠果冷榨油中不饱和脂肪酸的含量较高,可作为油脂工业原料。2、文冠果冷榨油的急性毒性试验结果表明,受试动物未发现明显中毒症状及死亡,按急性毒性分级标准判定文冠果油为无毒级别;且三项遗传毒性试验(Ames、骨髓细胞微核试验、小鼠精子畸变试验)结果均为阴性,说明文冠果冷榨油无毒性。3、文冠果冷榨油的30天喂养的亚急性毒性试验结果表明,试验SD大鼠未见明显的中毒症状及死亡。各剂量组指标与空白对照组比较,大鼠体重、食物利用率、血液学及脏/体比等指标均属于正常范围内,肉眼观察其宏观组织形态无异常现象。连续灌胃30天,动物受试物血液生化指标中,TP、ALB、ALT、AST、BUN、GLU与阴性对照组均无显著差异,TC、TG均在正常参考范围内;但各剂量组的CRE值均高于参考值范围,可能是文冠果油试验剂量偏高,对大鼠的肾脏可能有不良作用,需进一步研究确定。4、文冠果冷榨油的体外抗氧化活性表明,文冠果油的还原能力超过BHT、TBHQ;对邻二氮法产生的·OH的清除效果强于BHT、TBHQ;对DPPH具有很强的清除作用;对超氧阴离子的清除作用稍高于BHT、TBHQ;对Fe2+诱导脂蛋白PUFA过氧化反应表现出较好的抑制作用。且体内抗氧化活性表明,文冠果油可以显着提高受试小鼠肝组织、脑组织中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)及谷胱甘肽过氧化物酶(GSH-PX)的活性,而显著降低这些组织中丙二醛(MDA)的含量。因此,文冠果油具有显著的抗氧化性。

【Abstract】 Xanthoceras sorbifolia Bunge is the perennial deciduous small tree or bush of Xanthoceras Bunge of Sapindancae, which originates in the Loess Plateau in northern part of China and also named as MUGua, WENDengguo and SENDengmaodao. It has strong adaptability and is resistance to cold, drought, salt and alkali conditions, and is China’s rare and unique woody fuels and energy plants.The oil contents of X.sorbifolia Bunge seeds and kernel is 30.4% and 66.39% respectively. The unsaturated fatty acids content in its kernel oil is reach as high as 94%. So the X.sorbifolia kernel oil has good functions of softening the blood vessel and controling cardiovascular system disease, and has a better function in lowering cholesterol. The Xsorbifolia kernel oil can also be used as natural unsaturated factors or additives.The Xanthoceras oil has a longer history as food in the civil society, but the theoretical research for its edible security is still a blank, and the studies on its functional properties also is rare. In this paper, the cold mechanical pressing method is adopted to extract X.sorbifilia Bunge seed oil. The toxicological evaluation (acute toxicity, genetic toxicity tests, a 30-day feeding test) of Xanthoceras kernel oil is conducted to determine its security as food, and its functional properties was also studied to provide experimental evidence for the development of a new functional oil resource. The results of test are as follows:1. The cold-pressing extraction condition of Xsorbifilia oil were:the pressure ranges from 55 to 60 MPa, kernel/shell ratio 9:1, squeezing time lasts for 8h, the yield of oil get to 40.44%.The refractive index and relative density of Xsorbifilia oil was 1.4723 and 0.914 respectively, which was lower compared with other vegetable oils. Its acid value and peroxide value were 0.601 mgKOH/g and 0.16 meq/Kg, those indicated that X.sorbifilia cold-pressing oil was fresh and had no changes in its quality and the oil iodine value was as high as 121.4g/100g. So Xsorbifilia cold-pressing oil can be used as raw materials to eat with a high level of unsaturated fatty acids.2. The acute toxicity test of Xsorbifilia cold-pressing oil results showed that the test animals did not find obvious symptoms of poisoning and death, which manifests Xsorbifilia cold-pressing oil is non-toxic level according to the acute toxicity grading criteria. The three genetic toxicity tests (Ames, micronucleus test of mouse marrow cells and mouse sperm aberrance test) were all negative. 3.30-day feeding of sub acute toxicity test of X.sorbifilia cold-pressing oil results showed that SD rats had no obvious symptoms of poisoning and death. Comparing the dose group with the control group, animal weight, food utility, hematology, weight and coefficient of visceral organ and other indicators are within normal range. There is no macroscopic morphology anomaly observed with naked eyes. After 30d feeding test, compared with the negative control group, no significant difference of TP, ALB, ALT, AST, BUN, GLU of test animals’blood biochemical parameters were observed; CHO,TG were within the normal reference range. But each dose group’s CRE values were higher than the reference range, maybe Xanthoceras oil has adverse effects on rat kidneys and needs further study to determine.4. In vitro antioxidant activity test of X.sorbifilia cold-pressing oil manifests that the reduction capacity of Xanthoceras oil is stranger than BHT and TBHQ, its scavenging effect on-OH produced by O-nitrogen method is greater than BHT and TBHQ; the scavenging effect on superoxide anion is also slightly higher than that of BHT and TBHQ. It also has a strong scavenging effect of DPPH and showed good inhibition of Fe2+ induced lipoprotein peroxidation PUFA reaction.In vivo antioxidant activity test shows that Xanthoceras oil can significantly improve the subjected mice’s superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-PX) activity in their liver and brain tissue, and there is also a significant reduction in malonaldehyde(MDA) content of these tissues. In a word, Xanthoceras oil has significant anti-oxidation activity.

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