【作者】 隋小宇；

【导师】 杨磊；

【作者基本信息】 东北林业大学 ， 生药学， 2009， 硕士

【摘要】 本文以浙江产迷迭香为主要原料,研究目的是实现迷迭香的综合开发与利用。在提取精油后,分别对迷迭香中脂溶性和水溶性抗氧化成分进行分离、纯化,同时对迷迭香精油进行初步精制。并将得到的纯化产品采用DPPH法测定其抗氧化能力。研究的主要内容如下:1.综合比较了水中蒸馏、水蒸气蒸馏、同时蒸馏萃取等五种精油提取方法。结果表明,五种提取方法的粗精油产率分布在1.50-1.60m L/100g之间,水中蒸馏与无溶剂微波提取方法的粗精油产率相对较高。从提取所需时间上看,水蒸气蒸馏与无溶剂微波提取方法所用时间最短,均在1 h左右,而其他三种方法,均在2 h以上。研究还发现,无溶剂微波提取法具有良好的预处理效果。经过微波提取精油后的迷迭香叶片,其中的三种主要抗氧化剂成分鼠尾草酸、鼠尾草酚和迷迭香酸,均能在较长贮存时间内维持在较高的含量水平。而未经任何预处理的迷迭香鲜叶采摘后保存10天后,其中的鼠尾草酸、鼠尾草酚及迷迭香酸含量均显著降低。可见,无溶剂微波提取方法不仅提取精油所需时间短、产率高,而且还可以作为一种新鲜迷迭香原料贮存之前的预处理手段使用。2.建立了一种可靠稳定的迷迭香总酚酸含量的检测方法。最终确定的检测条件为:Folin-Ciocalteu试剂与10%Na₂CO₃的体积比为1:3,Folin-Ciocalteu试剂1.5 mL,显色温度25℃,反应时间2 h,检测波长765 nm。3.选用H1020大孔吸附树脂对迷迭香脂溶性总酚酸进行纯化,研究表明,树脂对迷迭香脂溶性总酚酸的吸附属于S型吸附。上样液质量浓度对树脂的吸附效率存在较大影响。当浓度为4.45 mg/mL的迷迭香脂溶性总酚酸提取液以1 mL/min的流速过柱,经动态纯化后,迷迭香脂溶性总酚酸的质量分数达到了70.46%。该纯化组分清除DPPH自由基的IC₅₀值为0.0469 mg/mL,与BHT接近。4.采用大孔吸附树脂纯化迷迭香中水溶性迷迭香酸,经实验证明,HPD-200L为理想的纯化材料。其饱和吸附量为25.27 mg/g,饱和吸附时间为3 h,体积分数为60%的乙醇溶液为适宜的解吸剂。当浓度为0.80 mg/mL的迷迭香酸提取液以1.0 mL/min的流速过柱,经动态纯化后,迷迭香脂溶性迷迭香酸的质量分数由7.98%提高到35.80%。该纯化组分清除DPPH自由基的IC₅₀值为0.0262 mg/mL,其抗氧化能力优于BHT。5.采用减压间歇精馏的方法对迷迭香粗精油进行初步精制。结果表明,当体系真空度为-0.095 MPa,采用变回流比方式,控制不同的温度区间进行产品收集,所得到的产品实现了初步精制的要求,其中的α-蒎烯、1,8-桉叶素、樟脑的含量分别达到88.17%、77.45%和78.82%。对精制产品的抗氧化活性研究表明,重油组分具有较好的抗氧化能力,说明通常作为残液处理的重油物质也具有很大的研究和利用前景。更多还原

【Abstract】 The purpose of the present study is to implement comprehensive development and utilization of rosemary, using rosmary planted in Zhejiang Province as main raw material. Isolating and purifying water-soluble and liposoluble antioxidant components from rosemary that enssential oils has been extracted, and preliminarily refining rosemary essential oil. In addition, determining antioxygenic property of purified product by using the 2,2-diphenyl-1-picrylhydrazyl （DPPH） radical scavenging method. The main research contents were as follows:1. Five extraction methods of essential oils were comprehensively compared, including water distillation、steam distillation and simultaneous distillation and extraction, etc. The results show that essential oils yield was distributed between 1.50 mL/100g and 1.60 mL/100g for five methods. Among them, water distillation and solvent-free microwave extraction had relatively higher yield. From aspect of extraction time, steam distillation and solvent-free microwave extraction need to spend about one hour, less than the other methods.It was also found that solvent-free microwave extraction showed a good pretreatment effect. Rosemary leaves that essential oils been extracted by microwave extraction, the three kinds of main antioxidative substance, carnosic acids carnosol and rosmarinic acid contents can be reserved for a longer time. Whereas the no-pretreatment fresh leaves’ carnosic acid、carnosol and rosmarinic acid contents reduced significantly after storing for about ten days. This shows that solvent-free microwave extraction is not only in short time and high efficiency, but also can be used as a kind of new pretreatment method of storing fresh material.2. A reliable and stable method is established to determine the content of total phenolic acids of rosemary by Folin-Ciocalteu colorimetry. The appropriate detection conditions were : ratio between the volume of Folin-Ciocalteu reagent and 10% Na₂CO₃ was 1:3 （v:v）; the volume of Folin-Ciocalteu reagent was 1.5 mL; reaction temperature was 25℃; reaction time was 2 h; the detection wavelength was 765 nm.3. H1020 resin was screened out from 9 kinds of macroporous resin,and it was the optimum macroporous resin to purify liposoluble total phenolic acids from rosemary. The results showed adsorption mode of H1020 resin belong to S style, sample concentration have a great influence on adsorption efficiency. The flow rate was 1mL/min when the concentration of rosemary total phenolic acids was 4.45mg/mL, After dynamic purification, the liposoluble total phenolic acids mass fraction increased from 47.74% to 70.46%, and its IC₅₀ value on scavenging DPPH radical was 0.0469 mg/mL, it was close to BHT.4. Study on the purification of rosmarinic acid from rosemary by macroporous resin. The results showed HPD-200L resin is a suitable material. Saturated adsorption capacity of HPD- 200L resin was 25.27 mg/g （dry resin）, saturated absorption time was 3 h, suitable desorption agent was 60% （volume fraction） ethanol solution, under the conditions of 20℃and pH 2-3, the flow rate was 1 mL/min when the concentration of rosmarinic acid was 0.8 mg/mL. After dynamic purification, the hydrosoluble rosmarinic acid mass fraction increased from 7.98% to 36.8%, and its IC₅₀ value on scavenging DPPH radical was 0.0262 mg/mL, it was superior to BHT.5. Preliminarily refine rosemary essential oil by vacuum batch distillation. The results showed acquired product reached the expected objective when system vacuum pressure was -0.095 MPa, using mode of changing reflux ratio and collecting different temperature section product. To obtain several product which content ofα-pinene、1,8-cineole and camphor reached to 88.17%、77.45% and 78.82% respectively. Research on antioxidative activity showed heavy oil has a better effect than others, therefore heavy oil fraction that is usually considered as discard raffinate will have a good prospect of research and application.更多还原