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1、硕士学位论文甘薯中薯蓣皂甙元的提取工艺研究论文作者：指导教师：教授副指导教师：申请学位类别：工学硕士学科、领域：食品科学所在单位：生物科学与工程学院答辩日期：年5月Hebei University of Science and TechnologyDissertation for the Master DegreeStudy on Extraction of Diosgenin from theSweet PotatoCandidate：Li XiaoqianSupervisor：Prof. Mou DehuaAssociate Supervisor：AcademicDegreeAppplie
2、d for：Master of EngineeringSpeciality：Food ScienceEmployer：College of Bioscience and BioengineeringDateofOralExamination:May, 2012l摘 要我国是世界上最大的甘薯生产国，相对于其他水果甘薯具有低脂、低热量和高纤维等优点，除了含有大量淀粉以外，还含有胡萝卜素、VB1、B2和VC，以及钙、钾、硒、铁等元素。近几年研究发现，甘薯中含有脱氢表雄酮，即DHEA，该物质具有多种保健功能，是循环系统中最为丰富的甾体激素之一，而据报道在植物体内DHEA以其前体物质薯蓣皂甙元的形式存在
3、，而薯蓣皂甙元作为一种重要的甾体中间体广泛的应用于医药工业生产。本课题主要针对河北省农科院甘薯组选育的新品种，进行甘薯中薯蓣皂甙元的提取工业研究；对检测方法进行对比优化；并对甘薯膨大期到成熟期薯蓣皂甙元在甘薯根、茎和叶中的含量进行了检测。为河北省新品种甘薯的推广种植和综合加工利用提供依据，并带动河北产业经济。本课题研究了分光光度法检测甘薯中薯蓣皂甙元的条件极其稳定性研究，通过对比三种显色剂，最终确定5%香草醛冰醋酸溶液+高氯酸溶液作为显色剂效果最佳，显色稳定条件为加入0.2 mL高氯酸，在80下水浴20 min，静置30 min。最终测得的薯蓣皂甙元特征吸收波长在540 nm。本课题考察了膨大
4、期甘薯中基础物质和薯蓣皂甙元的变化关系及相关性。通过试验可知，在膨大期随着淀粉含量逐渐增加，薯蓣皂甙元含量的增加趋势也越明显。块根中的薯蓣皂甙元含量随着生长周期的延长呈现明显的增加趋势，而薯皮中的薯蓣皂甙元在前期增长趋势明显，后期含量增加不明显，并且薯块中的薯蓣皂甙元含量明显高于薯皮。本课题采用了索氏提取法和超声波辅助方法对薯蓣皂甙元进行提取。两种方法比较而言，超声波辅助提取方法，具有提取率高，提取时间短，操作简单等优点，第三次提取率能达到97.54%。通过单因素和正交试验获得了超声波辅助提取的最优条件：酸浓度1 mol/L、酸解温度100、酸解时间3 h、pH值为5、溶剂提取时间1 h。最佳
5、提取条件下的二次提取率达到了92.80%。硅胶纯化甘薯中薯蓣皂甙元的粗提液最佳的洗脱溶剂为石油醚：乙酸乙酯=1：1，纯化后的样品纯化度能达到82.56%。RP-HPLC法定性定量检测纯化后的薯蓣皂甙元，并确定了最佳检测条件，流动相：乙腈：甲醇：水=8：1：1；流速：0.7 mL/min；进样量：20 L；检测波长：210 nm，检出时间为：15 min左右。关键词 甘薯；薯蓣皂甙元；分光光度法；提取；纯化 lAbstractChina is the worlds largest producer of sweet potato, relative to other fruits, sweet
6、potato have the advantages of low fat, low heat and high fiber, besides contains a lot of starch, but also contains beta-carotene, VB1, B2 and VC, and calcium, potassium, selenium, iron and other elements. Study found that in recent years, sweet potato contains DHEA, namely DHEA, the material has a
7、variety of health care function, is the most abundant in the circulation system of one of steroid hormones, and DHEA in plant body with its precursor diosgenin form existence,as a kind of important intermediates steroid widely used and the pharmaceutical industry production.This subject mainly aimed
8、 at the academy in hebei province of sweet potato varieties of breeding, sweet potato in the extraction of diosgenin industrial research; Compared to test method and optimization; And detect the diosgenin of sweet potato at swollen to maturity in each organ levels.For new varieties of sweet potato i
9、n hebei province cultivated and comprehensive processing using provides the basis, and drive the hebei industry economy.This topic research the spectrophotometry detection conditions and stability study of diosgenin, through comparing three show color agent, and finally determined that 5% vanillin g
10、lacial acetic acid solution-high chlorine acid solution as colour-display reagent, show color stability conditions for join 0.2 mL high chlorine acid, in 80 water under 20 min, and let rest for 30 min. Finally the measurement of diosgenin features absorption wavelength in the 540 nm.This subject res
11、earch sweet potato based and diosgenin at the swollen change relations and correlation. Starch content increase obviously, diosgenin the increase of the content of the more obvious trend. diosgenin content of root with the extension of the growth cycle is obviously increased, and diosgenin of potato
12、 skins only expands the period in the beginning and end of the growth trend obvious, and diosgenin of root was obviously higher than potato skins.This topic research the soxhlet method and utrasonic-assisted extraction method. Two methods in comparison, utrasonic-assisted extraction method, have hig
13、her extraction yield, extracting time short, simple operation etc, the third time extraction yield is 97.54%. By single factor and orthogonal test won the ultrasonic extraction auxiliary the optimal conditions: acid concentration 1 mol/L, acid solution temperature of 100, acid solution time 3 h, pH
14、value for 5,solvent extraction time 1 h. Best extraction conditions of secondary extraction yield to 92.80%.Silica gel purification diosgenin coarse extraction solvent the best elution liquid for petroleum ether:ethyl acetate= 1:1, purification of sample purification and could reach 93.14%. RP-HPLC
15、method to detect the sweet potato diosgenin best detection conditions for: mobile phase: acetonitrile: methanol: water=8:1:1; Flow rate: 0.7 mL/min; Into the sample weight: 20 L; Detected wavelength: 210 nm, detection for time:15 min.Key words Sweet potato；Diosgenin；Spectrophotometry；Extracted；Purificationl目 录摘 要IAbstractII第1章 绪 论11.1 甘薯简介11.2 甘薯主要营养成分简介11.3 薯蓣皂甙元的简介21.4 薯蓣皂甙元研究现状31.4.1 薯蓣皂甙元的功能特性31.4.2薯蓣皂甙元的提取现状41.4.3薯蓣皂甙元的纯化和检测方法的研究进展61.5 课题主要研究的内容、意义及创新点81.5.1 课题研究的内容和意义81.5.2 课题的创新点9第2章 甘薯中薯蓣皂甙元分光光度检测法102.1 试验材料与设备102.1.1 试验材料与试剂102.1.2 试验设备102.
