【作者】 刘辉；

【导师】 刘长江；

【作者基本信息】 沈阳农业大学 ， 食品科学与工程， 2009， 博士

【摘要】 芹菜籽为伞形花科草本植物芹菜（Apium graveolens Linn）的种子,在我国资源丰富,且尚未得到合理的有效利用。研究表明,芹菜籽中油脂含量丰富,含有多种药理活性成分,具有散气、消肿、利尿、抗肿瘤、抗衰老等功效。本文采用在我国产量最高的旱芹菜籽为试验原料,对其萃取出的芹菜籽油进行系统的研究,确定最佳的制备技术路线,并明确其生理活性,为更好的开发和利用芹菜籽资源提供理论依据。本文通过有机溶剂法、同时水蒸气蒸馏法和超临界流体萃取法研究芹菜籽油的提取工艺;采用分子蒸馏技术分离轻重组分纯化芹菜籽油;研究了芹菜籽油的理化性质和氧化稳定性,分析其营养成分;将芹菜籽油微胶囊化;通过动物实验评价芹菜籽油的毒理安全性,研究了芹菜籽油对高脂大鼠的血脂调节作用,并探讨其作用机理。主要研究结论如下:1、芹菜籽中粗脂肪含量达19.05%;含有19种基本氨基酸,总氨基酸含量230.569mg/g,8种必需氨基酸均含有,以谷氨酸含量最高;原子吸收光谱分析芹菜籽中矿物元素钾含量高达8.6mg/g,镁含量为1.0mg/g,钠、铁含量也比较丰富。2、有机溶剂法提取芹菜籽油最佳工艺条件为:粒度60目、液料比3∶1、浸泡48h、旋转蒸发温度50℃,提取率达11.47%。同时水蒸气蒸馏法最佳工艺条件为:粒度60目、液料比8∶1、浸泡时间12h、蒸馏时间8h,提取率为4.36%。超临界流体萃取法响应面优化工艺条件为:粒度60目、萃取压力21MPa、萃取温度46℃、CO₂流量16L/h、分离温度30℃、每次进料300g、萃取2h,工艺优化后芹菜籽油提取率达12.04%。3、分子蒸馏纯化芹菜籽油响应面法优化工艺条件为:进料温度50℃、蒸发温度111℃、刮膜转速214r/min、进料速率1.6mL/min,在此优化条件下重组分馏出物收率达71.08%。气质联用分析分子蒸馏后芹菜籽油主要成分和相对含量,共鉴定出占总成分91.6%的19种主要成分。轻组分化合物被分离在蒸出物中,而不饱和脂肪酸、丁基苯酞、瑟单内酯、角鲨烯等有效成分则分离在重组分馏出物中,且相对含量均有提高。4、芹菜籽油中不饱和脂肪酸含量丰富,不稳定。温度、时间、空气和光照可加速芹菜籽油的氧化速度。加入抗氧化剂可有效延缓氧化速度,四种抗氧化剂抗氧化性顺序为TBHQ＞PG＞BHT＞BHA,以抗坏血酸增效,抗氧化效果更好。与Cu²⁺、Fe³⁺两种金属离子接触,芹菜籽油过氧化值明显升高,Cu²⁺的影响更大。芹菜籽油应尽量在低温、隔绝空气、避光条件下保存,避免与金属离子的接触,为延长贮藏期可适当加入抗氧化剂。5、气相色谱法测定芹籽油中棕榈酸、硬脂酸、油酸、亚油酸,亚麻酸、花生酸组成及含量,在优化的色谱条件下,六种不饱和脂肪酸达到基线分离,结果表明芹菜籽油中棕榈酸的含量为92.86mg/g,硬脂酸含量为8.46mg/g,油酸含量为98.71mg/g,亚油酸的含量为283.24mg/g,亚麻酸的含量为19.69mg/g,花生酸的含量为34.28mg/g。反相高效液相色谱检测芹菜籽油中6-维生素E含量为86.9μg/g,γ-维生素E含量为120.6μg/g,β-维生素E含量为69.7μg/g,α-维生素E含量为132.5μg/g。6、以β-环糊精和明胶为复合壁材微胶囊化芹菜籽油的最佳壁材配方为:芯材占壁材含量24.70%,β-环糊精/明胶3.03倍,固形物浓度18.82%,乳化剂用量0.75%。以最优壁材配方包埋芹菜籽油,通过工艺优化模型得出最佳微胶囊化芹菜籽油的喷雾干燥工艺参数为32MPa压力下均质3次、进风温度181℃、出风温度80℃,在此条件下,芹菜籽油的微胶囊化效率达到92.11%,产率为84.46%。制备的微胶囊化芹菜籽油产品溶解性良好,模拟胃液中60min释放率可达90%以上,贮藏稳定性明显优于液态芹菜籽油,电镜扫描观察微胶囊结构均匀呈球形。红外光谱分析微胶囊化前后芹菜籽油的成分基本保留。7、芹菜籽油对小鼠经口LD₅₀＞10.00g/kg体重,Ames试验、小鼠骨髓微核试验结果均为阴性。所测芹菜籽油属实际无毒级,是安全的。8、与高脂模型组比,灌胃不同剂量芹菜籽油显著降低大鼠血清的总胆固醇（TC）、总甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）水平和载脂蛋白B（ApoB）含量,显著提高高密度脂蛋白胆固醇（HDL-C）水平和载脂蛋白A（ApoA I）含量,动脉硬化指数LDL-C/HDL-C显著下降;能有效防止高脂饮食引起的肝脏脂肪变性及细胞坏死,且呈一定剂量依赖关系;并使高脂血症大鼠血清和肝脏的SOD活性显著提高,脂质过氧化物MDA含量显著下降,且呈现一定的剂量依赖性。表明芹菜籽油能增强抗氧化酶活性,减少自由基的产生,防止脂质过氧化及其中间代谢产物对机体的损害,可以有效提高高脂血症大鼠体内的抗氧化水平,从而改善血脂代谢。综上所述,本论文系统的研究了制备芹菜籽油的工艺条件,分析芹菜籽油中的活性成分,明确了其降血脂的生理功能,为芹菜籽油的工业化生产提供切实可行的理论依据。更多还原

【Abstract】 Celery seed is the seed of umbelliferae herbage apium.There have abundant resources in our country.Celery seeds are not reasonable and effective to be used.Studies have shown that celery seeds contain abundant oil and pharmacological activities ingredients.Celery seeds have the efficacy on scatter chi,detumescence,diuresis,antitumor,anti-aging and etc.The test selected the celery seeds that had the most output in our country.Celery seeds oil（CSO） was systematic studied.It was very important that determined the technology pathway and its bioactivities to provide theoretical support for developing and utilizing the resources of celery seeds.In this paper,celery seeds oil was extracted by organic solvent extraction（SE）, simultaneous water distillation-solvent extraction（SDE）and supercritical carbon dioxide fluid extraction（SFE-CO₂）.The light component and weighty component were separated by molecular distillation purification.The physicochemical property and oxidative stability were tested.Nutrient content and ingredients were identified.Microencapsulation CSO was studied. The toxicological evaluation was completed.And the effects of CSO on hypolipidemic and mechanism in hyperlipidemic rats were studied by animal experiment.The main results were as follows:1.The content of crude fat from celery seeds was 19.05%.Celery seed contained 19 kind amino acids including 8 kind sessential amino acids.The content of total amino acid was 230.569mg/g,and the highest content was glutamic acid.Through the analysis by atomic absorption spectrum,the content of mineral elements K was 8.6mg/g,the content of Mg was l.0mg/g.In addition,the content of Na and Fe were plentiful.2.The optimum technical conditions by SE were:granularity was 60 mesh size,liquid seeds ratio was 3:1,soaking time was 48h,evaporation temperature was 50℃.Under these conditions,the extraction yield was 11.47%.The optimum technical conditions by SDE were: granularity was 60 mesh size,liquid seeds ratio was 8:1,soaking time was 12h,distilling time was 8h.Under the conditions,the extraction yield was 4.36%.The optimum response surface technical conditions by SE were:granularity was 60 mesh size,extraction pressure was 21MPa,extraction temperature was 46℃,flow rate of CO₂ was 16L/h,separation temperature was 30℃,incoming stock was 300g every time,extraction time was 2h.Under the optimize conditions,the extraction yield of CSO was 12.04%. 3.The response surface optimum technical conditions by molecular distillation were: charging temperature was 50℃,distillation temperature was 111℃,rotating speed was 214r/min,charging rate was 1.6mL/min.Under these optimize conditions,the yield of weighty constituents distillate was 71.08%.Through analyzing the main constituents of CSO by GC/MS,main components of 91.6%were identified,it included 19 kinds compound.The light constituents were separated as evaporant.The active ingredients including unsaturated fatty acid,NBP,Squalene and etc were separated in weighty constituent distillate.And the relative contents were increased.4.Celery seeds oil contained plentiful unsaturated fatty acid,it was instability.Some external conditions such as temperature,time,oxygen and light could accelerate the oxidation rate.The oxidation rate could defer by adding in some antioxidants.The sequence of inoxidability measurement was:TBHQ＞PG＞BHT＞BHA.The oxidation resistance effect was increased by adding ascorbic acid as synergistic agent.If celery seeds oil was in touch with Cu²⁺,Fe³⁺,the peroxide number would increase obviously.The influence of Cu²⁺was greater. Therefore,the CSO should be preserved at lower temperature,and should avoid touching oxygen,light,metallic ion.In order to prolonging the storage life of CSO,it should be appended right amount antioxidant.5.The content of unsaturated fatty acid including hexadecanoic acid,stearic acid,oleic acid,linoleic acid,linolenic acid,arachidic acid was determined by GC.The optimum chromatogram conditions,it achieved baseline separation.The content of hexadecanoic acid was 92.86mg/g,content of stearic acid was 8.46mg/g,content of oleic acid was 98.71mg/g, content of linoleic acid was 283.24mg/g,content of linolenic acid was 19.69mg/g,content of arachidic acid was 34.28mg/g.The content ofδ-V_E was 86.9μg/g,content ofγ-V_E was 120.6μg/g,content ofβ-V_e was 69.7μg/g,content ofα-V_E was 132.5μg/g.6.Glutin and p-CD were selected as complex wall material to embed celery seeds oil. The optimum prescription of wall material was:rate of core material to wall material was 24.70%,β-CD/glutin was 3.03,total solid concentration was 18.82%,content of emulsifier was 0.75%.The response surface optimum technical conditions of sponging drying were: homogeneity pressure was 32MPa,homogeneity times were there,Inlet air temperature was 181℃,Outlet air temperature was 80℃.Under the optimize conditions,the microencapsulation efficiency was 92.11%,the output was84.46%.The solubility of microcapsule product was well.The release rate in simulation gastric juice could attain 90% after 60min.Storge stability of microcapsule product excelled liquid state CSO.The microcapsule product was globose and its structure was well-distributed.Its ingredients were retained fundamentally by infrared spectrum analysis.7.The irritable toxicology test indicated LD₅₀＞10.00g/kg.The results of Ames test and micronucleus test of bone marrow cell were feminine.So the CSO belonged to innocuity grade.8.The serum TG,TC,LDL-C concentration,ApoB content and atherosclerosis index in rats fed with CSO were significantly lower than those in model group,while serum HDL-C concentration and ApoA I content were higher.CSO could prevent steatosis and cytoclasis causing by high fat diet,and it had dose-dependent.Comparing with model group,SOD activity were increased significantly in serum and liver by feeding with CSO.MDA content were decreased in serum and liver.CSO could increase antioxidase activity,decrease the freeradical.prevent the harm of lipid peroxide and intermediary metabolism.CSO could improve oxidation resistance level of hyperlipemia rats and regulate blood lipid metabolism.As we had seen,the test studied systematically the manufacture technology condition of CSO,analysised its bioactivities ingredients,and determined the hypolipidemic function.The study could provide theoretical support for industrial production of CSO.更多还原