【作者】 姜兴华；

【导师】 范青生；

【作者基本信息】 南昌大学 ， 食品科学， 2012， 硕士

【摘要】 五味子为木兰科五味子(Schisandra chinensis (Turcz.) Baill,俗称北五味子)或华中五味子(S.sphenanthera Rehd.et Wils,俗称南五味子)的干燥成熟果实,五味子已有上千年的药用历史,药用价值极高。现代药理学研究表明五味子的生物活性成分木脂素,尤其是联苯环辛烯类木脂素,具有护肝、镇静催眠、抗氧化应激、促进血管舒张、抗炎、抗癌以及增强免疫力等作用。本研究建立了一种同时检测五味子中五味子醇甲、五味子醇乙、五味子甲素、五味子乙素和五味子丙素5种有效成分的高效液相色谱法,优化了五味子功能饮料的制备工艺并考察了其稳定性,同时对五味子保健葡萄酒的制备工艺进行了研究并对其进行了功能评价。主要研究及结果如下：经紫外分光光度计扫描确定五味子醇甲、五味子醇乙、五味子甲素、五味子乙素和五味子丙素同时检测的最佳波长为254nm,建立的HPLC检测方法为：色谱柱：KromasilC18柱(250×4.6nm)；波长：254nm；流速：1.0mL/min;柱温：30℃；试样温度：室温；进样量：20μL；流动相：甲醇(A)和水(B)进行梯度洗脱,0-8min30%B,8-12min30%-20%B,12-20min20%-10%B,20-30min10%B。并对其线性范围、精密度、稳定性、重复性和加标回收率进行了考察。结果表明该方法精确可靠、重复性好,可用于测定五味子和含五味子的保健食品中的有效成分含量。对五味子的提取工艺进行了研究,采用正交试验设计分别考察了料液比、温度、提取时间、提取次数及醇浓度对五味子木脂素提取率的影响。通过正交实验,优化得到水提法的最佳工艺条件是：料液比1：12,温度50℃,提取时间2h,提取3次；醇提法的最佳工艺条件是：料液比1：12,醇浓度80%,提取温度90℃,提取时间2h,提取3次。同时对比了醇提水沉法与水提法和醇提法最佳工艺的五味子醇甲得率和5种木脂素总得率,结果表明醇提法提取所得五味子醇甲含量和5种木脂素总含量均高于水提法和醇提水沉法,而水提法和醇提水沉法两种方法所得提取物中五味子醇甲含量差别不大。因此五味子功能饮料的工艺中选用水提法,而五味子保健葡萄酒的制备工艺中选用醇提法。探究了五味子功能饮料的制备工艺,干燥成熟的五味子果实经提取、过滤、澄清、掩蔽和调配可得五味子功能饮料。经实验优化得到最佳澄清工艺为：2%壳聚糖溶液作为最佳澄清剂,每100mL的五味子水提物(生药量为0.02g/mL)添加2%壳聚糖溶液2mL,澄清条件为室温下静置1h。最佳掩蔽包合工艺为：以0.01g/mL的五味子提取液为基准,按1.6g/100mL添加β-环糊精,在50℃下搅拌80min。甜菊糖和蜂蜜添加量为饮料质量的0.8‰和2%(以0.01g/mL的五味子提取液为基准)。同时对五味子功能饮料进行了稳定性考察,结果表明经强光照射和加速实验后,五味子功能饮料性状、pH、澄清度和功效成分含量并无明显变化。研究了五味子保健葡萄酒的制备工艺,干燥成熟的五味子果实经提取、过滤、浓缩、精滤和调配等步骤可得五味子保健葡萄酒。该五味子保健葡萄酒符合GB15037-2006《葡萄酒》和GB16740-1997《保健(功能)食品通用标准》的规定。考察了五味子保健葡萄酒的保健作用,以8.3mL/kg·bw、16.7mL/kg·bw、50.0mL/kg·bw剂量的五味子保健葡萄酒经口灌胃给予小鼠30天。50.0mL/kg·bw剂量能延长阀剂量戊巴比妥钠诱导的小鼠睡眠时间、缩短巴比妥钠睡眠潜伏期,与空白和酒基对照组比较差异有显著性(P<0.05),表明五味子保健葡萄酒具有改善睡眠的功能；16.7mL/kg·bw、50.0mL/kg·bw剂量能提高小鼠迟发型变态反应能力、抗体生成细胞数和NK细胞活性,与空白和酒基对照组比较差异有显著性(P<0.05或P<0.01),表明五味子保健葡萄酒具有增强免疫力的功能。更多还原

【Abstract】 The drug Wuweizi is dried fruits of Schisandra chinensis or S. sphenantherd, it has been used for thousands of years and its officinal value is high. Modern pharmacological research has demonstrated that most of the biological actions and pharmacological effects of Wuweizi can be attributed to its lignan constituents, particularly the dibenzocyclooctadiene-type lignans, which show protection for liver, sedative hypnotic, protection against oxidative stress, vasodilative, anti-inflammatory, anti-proliferative and enhancing immunity effects.A simple, rapid and specific HPLC method was established for simultaneous determination of five major lignans (Schisandrol A, Schisandrol B, Schisandrin A, y-Schisandrin and Schisandrin C) in Schisandra chinensis. The preparation technology and stability of functional drink of Schisandra chinensis were researched. The preparation technology of health-care wine of Schisandra chinensis was studied and health benefits of the wine were evaluated. The major results as follow:The best detection wavelength was identified as254nm after the five lignans scanned via ultraviolet spectrophotometer, and the five lignans could be separated completely on Kromasil C18column (250X4.6nm) and then detected at254nm, using methanol (mobile phase A) and water (mobile phase B) with gradient elution as the mobile phase at flow rate1.0mL/min, and the column temperature was30℃. Mobile phase B kept30%(V/V) in the first8minutes, and decreased linearly from30%to20%within4minutes, then decreased linearly again from20%to10%within8minutes, and kept constant to30th minute, finally increased linearly from10%to30%within15minutes. The method was validated through linearity, precision, stability, repeatability and recovery, the results showed that the method is accurate and reproducible. This HPLC method established is suitable for routine quantitative analysis of Schisandra chinensis and multi-herb remedies and healthy food containing this herb.The extraction technology of water extraction and alcohol reflux method was optimized, the effects of the ratio of material to solution, ethanol solution concentration, temperature, extracting time and extracting times on the yields of Schisandrol A and five lignans were tested by orthogonal experiment. The optimal conditions of water extraction and alcohol reflux method were as following respectively:the ratio of material to solution1:12, temperature50℃, extracting time2hour and extracting times3times; the ratio of material to solution1:12, ethanol concentration87%, temperature90℃, extracting time2hour and extracting times3times. And the yields of Schisandrol A and five lignans obtained by alcohol reflux method, alcohol extraction-water precipitation method and water extraction method were compared. The results showed that the yields of Schisandrol A and five lignans obtained by alcohol reflux method were high than another two methods. And the difference is not significant between alcohol extraction-water precipitation method and water extraction method. So water extraction method and alcohol reflux method were respectively selected to extract lignans for functional drink and health-care wine of Schisandra chinensis preparation.Preparation technology of functional drink of Schisandra chinensis was studied, the drink was prepared through the process of extraction, filtration, clarification, masking, allocation and so on from dried ripe fruits of Schisandra chinensis. The optimal clarification conditions were as following:100mL aqueous extract of Schisandra chinensis (0.02g/mL) was added2mL2%chitosan solution, then let the solution still stand at room temperature for1hour. The optimal masking conditions were as following:100mL the aqueous extract of Schisandra chinensis (0.01g/mL) was added1.6gβ-CD, then the solution was stirred at50℃for80minutes. Finally honey and stevioside, of which the qualities were0.8%o and2%respectively of the drink (0.01g/mL) were added. The paper researched the stability of the functional drink of Schisandra chinensis, the results showed that the appearance, pH, clarity and the content of functional composition of the functional drink of Schisandra chinensis were not changed significantly after high-light exposure experiment and accelerating experiment.Preparation technology of health-care wine of Schisandra chinensis was researched, the wine was prepared through the process of extraction, filtration, concentration, pure filtration, allocation and so on from dried ripe fruits of Schisandra chinensis. The health-care wine conforms to the regulations of GB15037-2006and GB16740-1997.This paper also researched health benefits of the health-care wine. The health-care wine was given to mice through mouth at three doses (8.3mL/kg·bw、16.7mL/kg-bw、50.0mL/kg-bw) for30days. There was significant difference (P<0.05) between50.0mL/kg-bw dosage of the sample and the blank and wine-based control group, respectively, which could significantly extend the sleeping time of tested mice induced by Sodium Pentobarbital, significantly shorten incubation period induced by Sodium Pentobarbital, so the results showed that the health-care wine of Schisandra chinensis can improve sleep. And there was significant difference (P<0.05or P<0.01) between16.7mL/kg·bw,50.0mL/kg·bw dosage of the sample and the blank and wine-based control group, respectively, which could significantly enhance the ability of delayed hypersensitivity, the quantity of antibody producing cell, activity of natural killer cell, so the results showed that the health-care wine of Schisandra chinensis can boost immunity.更多还原