五味子功能饮料及保健葡萄酒的研制
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。
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.
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。
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.
引文
[1]Ip S, Ko K. The crucial antioxident action of schisandrin B in protecting against carbon tetrachloride hepatotoxicity in mice:A comparative study with butylated hydroxytoluene[J]. Biochemical Pharmacology.1996,52:1687-1693.
[2]Huang F, Xiong Y T, Xu L H, et al. Sedative and hypnotic activities of the ethanol fraction from Fructus Schisandrae in mice and rats[J]. Journal Of Ethnopharmacology.2007,110(3): 471-475.
[3]Lam P Y, Yan C W, Chiu P Y, et al. Schisandrin B protects against solar irradiation-induced oxidative stress in rat skin tissue[J]. Fitoterapia.2011,82(3):393-400.
[4]Lam P Y, Chiu P Y, Leung H Y, et al. Schisandrin B co-treatment ameliorates the impairment on mitochondrial antioxidant status in various tissues of long-term ethanol treated rats[J]. Fitoterapia.2010,81(8):1239-1245.
[5]Park J Y, Shin H K, Lee Y J, et al. The mechanism of vasorelaxation induced by Schisandra chinensis extract in rat thoracic aorta[J]. Journal of Ethnopharmacology.2009,121:69-73.
[6]Guo L Y, Hung T M, Bae K H, et al. Anti-inflammatory effects of schisandrin isolated from the fruit of Schisandra chinensis Baill[J]. European Journal Of Pharmacology.2008,591:293-299.
[7]孙杰.五味子乙素预防阿霉素引起的慢性心肌毒性并增强其抗肿瘤活性的体内研究[D].浙江大学,2010.
[8]Min H, Park E, Hong J, et al. Antiproliferative effects of dibenzocyclooctadiene lignans isolated from Schisandra chinensis in human caneer cells[J]. Bioorganic & Medicinal Chemistry letters.2008,18:523-526.
[9]仰榴青,陈荣华,吴向阳,等.五味子醇提残渣中粗多糖的免疫活性研究[J].食品科学.2008(6)：392-394.
[10]He X G, Lian L Z, Lin L Z. Analysis of lignan constituents from Schisandra chinensis by liquid chromatography-electrospray mass spectrometry[J]. Journal Of Chromatography A.1997, 757(1-2):81-87.
[11]周英,杨峻山,王立为,等.五味子科植物三萜成分[J].中国药学杂志.2003(2)：81-83.
[12]Wang L, Chen Y, Song Y, et al. GC-MS of volatile components of Schisandra chinensis obtained by supercritical fluid and conventional extraction[J]. JOURNAL OF SEPARATION SCIENCE.2008,31:3238-3245.
[13]赵婷,仰榴青,笪祖林,等.五味子醇提残渣中多糖的提取工艺研究[J].食品研究与开发.2008(8)：70-73.
[14]范荣军,任涛,刘成柏,等.五味子多糖提取工艺的比较研究[J].中成药.2008(6)：827-831.
[15]张兰杰,张维华,赵珊红.北五味子果实中多糖的提取与纯化研究[J].鞍山师范学院学报.2002(1)：58-60.
[16]金鑫,辛爱学.南五味子提取工艺研究[J].中国现代药物应用.2009(6)：149-151.
[17]田振坤,王春晖,陈忠新.高效液相色谱法优选五味子浸膏的提取工艺[J].中医药学报.2009(2)：43-45.
[18]马永全,于新,黄雪莲.响应面法优化南五味子甲素提取工艺的研究[J].食品科技. 2010(12)：206-209.
[19]邢军,李正国.均匀设计法优选五味子药材水提取工艺[J].中国药业.2008(10)：51-52.
[20]戴军,徐佐旗,赵婷,等.超临界CO2提取五味子木脂素的工艺研究[J].食品与药品.2010(9)：312-315.
[21]顾蔚,李宗霖,王喆之.华中五味子中木脂素提取工艺研究[J].中药材.2010(9)：1501-1503.
[22]杨磊,马春慧,祖元刚.北五味子木脂素超声提取工艺[J].农业工程学报.2009(S1)：185-192.
[23]Lee H J, Kim C Y. Simultaneous determination of nine lignans using pressurized liquid extraction and HPLC-DAD in the fruits of Schisandra chinensis[J]. Food Chemistry.2010,120(4): 1224-1228.
[24]Wang B L, Hu J P, Tan W, et al. Simultaneous quantification four active schisandra lignans from a traditional Chinese medicine Schisandra chinensis (Wuweizi) in rat plosma using liquid chromatography-mass apectrometry[J]. Journal of Chromatography B.2008,865:114-120.
[25]Zhang H, Zhang G Q, Zhu Z Y, et al. Determination of six lignans in Schisandra chinensis (Turcz.) Baill. Fruits and related Chinese multiherb remedies by HPLC[J]. Food Chemistry.2009, 115(2):735-739.
[26]窦志华,丁安伟,罗琳,等.复方五仁醇胶囊含药血清中五味子醇甲、五味子醇乙、五味子甲素、五味子乙素的测定[J].中草药.2009(3)：386-389.
[27]李丽波,高涵,孙超,等.五味子乙素诱导的HSP27和HSP70对小鼠肝损伤的保护作用[J].齐齐哈尔医学院学报.2010(8)：1177-1178.
[28]M. Zhu K F L, Yeung R Y, Li R C. Evalution of the protective efforts of Schisandra chinensis on Phase I drug metabolism using a CC14 intoxication model[J]. Journal of Ethnopharmacology.1999,67:61-68.
[29]孔华丽,闫亮,段惠娟.五味子醇提取物保肝作用成分分析[J].解放军药学学报.2010(1)：27-29.
[30]余凌虹,刘耕陶.五味子联苯环辛烯类木脂素成分的结构与药理活性关系和药物创新[J].化学进展.2009(1)：66-76.
[31]Panossian A, Wikman G. Pharmacology of Schisandra chinensis Bail.:An overview of Russian research and uses in medicine[J]. Journal of Ethnopharmacology.2008,118:183-212.
[32]林蔚,黄宗锈,陈冠敏,等.中药五味子改善小鼠睡眠作用的研究[J].海峡预防医学杂志.2009(4)：51-52.
[33]徐亚杰,刘雅静,刘彤.考察北五味子不同提取工艺对镇静催眠作用的影响[J].山西医药杂志.2009(11)：1056.
[34]王雯雯,仰榴青,李永金,等.南、北五味子提取物对小鼠镇静、催眠作用的影响[J].江苏大学学报(医学版).2008(2)：122-123.
[35]徐丽华,黄芳,孙萌,等.南北五味子镇静催眠活性部位共有成分的分析[J].分析化学.2009(6)：828-834.
[36]黄莉莉,李廷利,郭冷秋,等.五味子对自由活动大鼠睡眠时相的影响[J].中药药理与临床.2007(5)：126-127.
[37]钮心懿,王维君,边振甲,等.五味子有效成分“醇甲”的中枢神经系统作用[J].药学学报.1983(6)：416-421.
[38]王文燕,邢晓冬,陈建光.北五味子提取液对高脂血症大鼠调脂作用的研究[J].中国老年学杂志.2009(21).
[39]高文荣,金香子,李香丹.五味子水提液对D-半乳糖所致衰老神经细胞的保护作用[J].时珍国医国药.2010(10)：2705-2706.
[40]裴文芳,单安山,石莉莎,等.五味子CO2超临界提取物对肉仔鸡免疫功能的影响[J].东北农业大学学报.2009(9)：70-74.
[41]李建平,单安山,程宝晶,等.五味子、柠檬酸对断奶仔猪血清生化指标和免疫功能的影响[J].中国畜牧杂志.2009(17)：25-29.
[42]韩占臣,张永泉.五味子提取物对肥育猪生产性能和免疫功能的影响[J].黑龙江畜牧兽医.2010(8)：79-81.
[43]黄秀兰,朱家媛,胡宏,等.五味子水煎剂对小鼠免疫功能的影响[J].川北医学院学报.1997(3)：9-10.
[44]刘力生,王勤,郑荣梁,等.五味子乙素对小鼠免疫功能、腹水型肝癌细胞DNA合成和存活率的作用[J].兰州大学学报.1990(1)：55-59.
[45]马永全,刘福瑜,于新.南药五味子提取物清除自由基活性的研究[J].食品工业.2010(3)：1-4.
[46]王昌.复方五味子护肝保健饮料的研制及其功能性评价的研究[D].黑龙江八一农垦大学,2010.
[47]文连奎,张久丽,姜明珠,等.北五味子露酒加工工艺研究[J].食品科学.2005(9)：278-280.
[48]田颖.五味子保健饮料配方优选研究[J].饮料工业.2007(10)：12-14.
[49]赵海智,李国玉,白天红.五味子果浆对乳酸菌生长的影响及五味子酸奶的加工[J].中国乳品工业.2003(4)：28-30.
[50]黄宏星.中国6城市普通人群失眠状况及京沪穗医师对抑郁伴失眠患者的认知及处理状况调查[J].中国医药导刊.2006(2)：100.
[51]陆明,王旭,朱壵,等.江苏地区不同年龄层人群失眠相关情况调查[J].深圳中西医结合杂志.2010(3)：173-176.
[52]老年人容易患癌症与免疫力低下有关[J].
[53]曹元成山东省淄博市中医院主任医师.免疫力下降莫忽视[N].健康报.
[54]魏强.高效液相色谱分析三类保健食品中常见违禁药物的方法研究[D].南昌大学,2008.
[55]耿立超,杨中林HPLC法测定不同产地五味子中木脂素的含量[J].中医药学报.2007(2)：47-48.
[56]Halstead C W, Lee S, Khoo C S, et al. Validation of a method for the simulaneous determination of four kings of lignans in the raw herb and commercial dried aqueous extracts of Schisandra chinensis by RP-LC with DAD[J]. Journal of Pharmaceutical And Biomedical Analysis.2007,45:30-37.
[57]马永全,黄雪莲,于新.响应面法优化五味子提取工艺研究[J].广东农业科学.2010(9)：151-153.
[58]高妮,龚又明,刘利根.牛蒡子提取液的澄清工艺研究[J].海峡药学.2006(4)：24-25.
[59]邹宇晓,廖森泰,刘学铭,等.澄清型金桑菊凉茶饮料工艺研究[J].食品研究与开发.2010(1)：78-80.
[60]姚艾东,颜栋美,李燕.山楂汁澄清工艺的研究[J].饮料工业.2001(2)：32-35.
[61]张卫国,郭红辉,刘欣.明矾澄清灵芝饮料效果的研究[J].广州食品工业科技.2003(3)：27-28.
[62]杨宏图,常翠,饶曼妮,等.五味子宁神口服液澄清工艺改进[J].中国药师.2006(11)：1065-1066.
[63]吴丹,薛旭初,马子骏,等.几种澄清剂在水蜜桃发酵酒中的应用[J].食品工业.2006(4)：11-13.
[64]谭勇,刘四新,柏玉娜,等.几种澄清剂对腰果梨酒品质的影响[J].食品科学.2009(21)：21-23.
[65]张陈云,刘金福,王娜.不同澄清剂对冬枣酒澄清效果的研究[J].食品科技.2010(8)：134-137.
[66]吴国卿,王文平,田亮,等.不同澄清剂对野木瓜果汁澄清效果的影响[J].贵州农业科学.2010(7)：171-172.
[67]陈娟,阚建全,杨蓉生,等.蜂蜜桑椹酒的澄清研究[J].食品工业科技.2010(6)：203-206.
[68]苏槟楠,谢红,刘春卉,等.低糖型金耳饮料口感调配研究初探[J].太原科技.2003(6)：75-76.
[69]刘春凤,郑飞云,李永仙,等.啤酒口感品评的模糊综合评价法[J].食品科学.2008(4)：138-142.
[70]刘春凤,董建军,郑飞云,等.模糊综合评价法在啤酒口感协调性品评中的应用[J].西北农林科技大学学报(自然科学版).2008(3)：213-222.
[71]乔海鸥,丁晓雯,张庆祝,等.柑桔类果汁的脱苦技术[J].包装与食品机械.2003(5)：27-31.
[72]左安连,毛海舫,李琼,等.柑橘类果汁脱苦方法研究综述[J].香料香精化妆品.2008(3)：33-39.
[73]梁泽建.柑桔汁脱苦技术的研究[D].四川农业大学,2007.
[74]潘利华,徐迪,布文婕.柑橘类果汁脱苦的研究进展[J].饮料工业.2006(2)：6-9.
[75]徐国胜,潘利华,杨阳,等.β-环糊精对柑橘类果汁脱苦效果的研究[J].安徽农业科学.2006(20)：5366-5367.
[2]Huang F, Xiong Y T, Xu L H, et al. Sedative and hypnotic activities of the ethanol fraction from Fructus Schisandrae in mice and rats[J]. Journal Of Ethnopharmacology.2007,110(3): 471-475.
[3]Lam P Y, Yan C W, Chiu P Y, et al. Schisandrin B protects against solar irradiation-induced oxidative stress in rat skin tissue[J]. Fitoterapia.2011,82(3):393-400.
[4]Lam P Y, Chiu P Y, Leung H Y, et al. Schisandrin B co-treatment ameliorates the impairment on mitochondrial antioxidant status in various tissues of long-term ethanol treated rats[J]. Fitoterapia.2010,81(8):1239-1245.
[5]Park J Y, Shin H K, Lee Y J, et al. The mechanism of vasorelaxation induced by Schisandra chinensis extract in rat thoracic aorta[J]. Journal of Ethnopharmacology.2009,121:69-73.
[6]Guo L Y, Hung T M, Bae K H, et al. Anti-inflammatory effects of schisandrin isolated from the fruit of Schisandra chinensis Baill[J]. European Journal Of Pharmacology.2008,591:293-299.
[7]孙杰.五味子乙素预防阿霉素引起的慢性心肌毒性并增强其抗肿瘤活性的体内研究[D].浙江大学,2010.
[8]Min H, Park E, Hong J, et al. Antiproliferative effects of dibenzocyclooctadiene lignans isolated from Schisandra chinensis in human caneer cells[J]. Bioorganic & Medicinal Chemistry letters.2008,18:523-526.
[9]仰榴青,陈荣华,吴向阳,等.五味子醇提残渣中粗多糖的免疫活性研究[J].食品科学.2008(6)：392-394.
[10]He X G, Lian L Z, Lin L Z. Analysis of lignan constituents from Schisandra chinensis by liquid chromatography-electrospray mass spectrometry[J]. Journal Of Chromatography A.1997, 757(1-2):81-87.
[11]周英,杨峻山,王立为,等.五味子科植物三萜成分[J].中国药学杂志.2003(2)：81-83.
[12]Wang L, Chen Y, Song Y, et al. GC-MS of volatile components of Schisandra chinensis obtained by supercritical fluid and conventional extraction[J]. JOURNAL OF SEPARATION SCIENCE.2008,31:3238-3245.
[13]赵婷,仰榴青,笪祖林,等.五味子醇提残渣中多糖的提取工艺研究[J].食品研究与开发.2008(8)：70-73.
[14]范荣军,任涛,刘成柏,等.五味子多糖提取工艺的比较研究[J].中成药.2008(6)：827-831.
[15]张兰杰,张维华,赵珊红.北五味子果实中多糖的提取与纯化研究[J].鞍山师范学院学报.2002(1)：58-60.
[16]金鑫,辛爱学.南五味子提取工艺研究[J].中国现代药物应用.2009(6)：149-151.
[17]田振坤,王春晖,陈忠新.高效液相色谱法优选五味子浸膏的提取工艺[J].中医药学报.2009(2)：43-45.
[18]马永全,于新,黄雪莲.响应面法优化南五味子甲素提取工艺的研究[J].食品科技. 2010(12)：206-209.
[19]邢军,李正国.均匀设计法优选五味子药材水提取工艺[J].中国药业.2008(10)：51-52.
[20]戴军,徐佐旗,赵婷,等.超临界CO2提取五味子木脂素的工艺研究[J].食品与药品.2010(9)：312-315.
[21]顾蔚,李宗霖,王喆之.华中五味子中木脂素提取工艺研究[J].中药材.2010(9)：1501-1503.
[22]杨磊,马春慧,祖元刚.北五味子木脂素超声提取工艺[J].农业工程学报.2009(S1)：185-192.
[23]Lee H J, Kim C Y. Simultaneous determination of nine lignans using pressurized liquid extraction and HPLC-DAD in the fruits of Schisandra chinensis[J]. Food Chemistry.2010,120(4): 1224-1228.
[24]Wang B L, Hu J P, Tan W, et al. Simultaneous quantification four active schisandra lignans from a traditional Chinese medicine Schisandra chinensis (Wuweizi) in rat plosma using liquid chromatography-mass apectrometry[J]. Journal of Chromatography B.2008,865:114-120.
[25]Zhang H, Zhang G Q, Zhu Z Y, et al. Determination of six lignans in Schisandra chinensis (Turcz.) Baill. Fruits and related Chinese multiherb remedies by HPLC[J]. Food Chemistry.2009, 115(2):735-739.
[26]窦志华,丁安伟,罗琳,等.复方五仁醇胶囊含药血清中五味子醇甲、五味子醇乙、五味子甲素、五味子乙素的测定[J].中草药.2009(3)：386-389.
[27]李丽波,高涵,孙超,等.五味子乙素诱导的HSP27和HSP70对小鼠肝损伤的保护作用[J].齐齐哈尔医学院学报.2010(8)：1177-1178.
[28]M. Zhu K F L, Yeung R Y, Li R C. Evalution of the protective efforts of Schisandra chinensis on Phase I drug metabolism using a CC14 intoxication model[J]. Journal of Ethnopharmacology.1999,67:61-68.
[29]孔华丽,闫亮,段惠娟.五味子醇提取物保肝作用成分分析[J].解放军药学学报.2010(1)：27-29.
[30]余凌虹,刘耕陶.五味子联苯环辛烯类木脂素成分的结构与药理活性关系和药物创新[J].化学进展.2009(1)：66-76.
[31]Panossian A, Wikman G. Pharmacology of Schisandra chinensis Bail.:An overview of Russian research and uses in medicine[J]. Journal of Ethnopharmacology.2008,118:183-212.
[32]林蔚,黄宗锈,陈冠敏,等.中药五味子改善小鼠睡眠作用的研究[J].海峡预防医学杂志.2009(4)：51-52.
[33]徐亚杰,刘雅静,刘彤.考察北五味子不同提取工艺对镇静催眠作用的影响[J].山西医药杂志.2009(11)：1056.
[34]王雯雯,仰榴青,李永金,等.南、北五味子提取物对小鼠镇静、催眠作用的影响[J].江苏大学学报(医学版).2008(2)：122-123.
[35]徐丽华,黄芳,孙萌,等.南北五味子镇静催眠活性部位共有成分的分析[J].分析化学.2009(6)：828-834.
[36]黄莉莉,李廷利,郭冷秋,等.五味子对自由活动大鼠睡眠时相的影响[J].中药药理与临床.2007(5)：126-127.
[37]钮心懿,王维君,边振甲,等.五味子有效成分“醇甲”的中枢神经系统作用[J].药学学报.1983(6)：416-421.
[38]王文燕,邢晓冬,陈建光.北五味子提取液对高脂血症大鼠调脂作用的研究[J].中国老年学杂志.2009(21).
[39]高文荣,金香子,李香丹.五味子水提液对D-半乳糖所致衰老神经细胞的保护作用[J].时珍国医国药.2010(10)：2705-2706.
[40]裴文芳,单安山,石莉莎,等.五味子CO2超临界提取物对肉仔鸡免疫功能的影响[J].东北农业大学学报.2009(9)：70-74.
[41]李建平,单安山,程宝晶,等.五味子、柠檬酸对断奶仔猪血清生化指标和免疫功能的影响[J].中国畜牧杂志.2009(17)：25-29.
[42]韩占臣,张永泉.五味子提取物对肥育猪生产性能和免疫功能的影响[J].黑龙江畜牧兽医.2010(8)：79-81.
[43]黄秀兰,朱家媛,胡宏,等.五味子水煎剂对小鼠免疫功能的影响[J].川北医学院学报.1997(3)：9-10.
[44]刘力生,王勤,郑荣梁,等.五味子乙素对小鼠免疫功能、腹水型肝癌细胞DNA合成和存活率的作用[J].兰州大学学报.1990(1)：55-59.
[45]马永全,刘福瑜,于新.南药五味子提取物清除自由基活性的研究[J].食品工业.2010(3)：1-4.
[46]王昌.复方五味子护肝保健饮料的研制及其功能性评价的研究[D].黑龙江八一农垦大学,2010.
[47]文连奎,张久丽,姜明珠,等.北五味子露酒加工工艺研究[J].食品科学.2005(9)：278-280.
[48]田颖.五味子保健饮料配方优选研究[J].饮料工业.2007(10)：12-14.
[49]赵海智,李国玉,白天红.五味子果浆对乳酸菌生长的影响及五味子酸奶的加工[J].中国乳品工业.2003(4)：28-30.
[50]黄宏星.中国6城市普通人群失眠状况及京沪穗医师对抑郁伴失眠患者的认知及处理状况调查[J].中国医药导刊.2006(2)：100.
[51]陆明,王旭,朱壵,等.江苏地区不同年龄层人群失眠相关情况调查[J].深圳中西医结合杂志.2010(3)：173-176.
[52]老年人容易患癌症与免疫力低下有关[J].
[53]曹元成山东省淄博市中医院主任医师.免疫力下降莫忽视[N].健康报.
[54]魏强.高效液相色谱分析三类保健食品中常见违禁药物的方法研究[D].南昌大学,2008.
[55]耿立超,杨中林HPLC法测定不同产地五味子中木脂素的含量[J].中医药学报.2007(2)：47-48.
[56]Halstead C W, Lee S, Khoo C S, et al. Validation of a method for the simulaneous determination of four kings of lignans in the raw herb and commercial dried aqueous extracts of Schisandra chinensis by RP-LC with DAD[J]. Journal of Pharmaceutical And Biomedical Analysis.2007,45:30-37.
[57]马永全,黄雪莲,于新.响应面法优化五味子提取工艺研究[J].广东农业科学.2010(9)：151-153.
[58]高妮,龚又明,刘利根.牛蒡子提取液的澄清工艺研究[J].海峡药学.2006(4)：24-25.
[59]邹宇晓,廖森泰,刘学铭,等.澄清型金桑菊凉茶饮料工艺研究[J].食品研究与开发.2010(1)：78-80.
[60]姚艾东,颜栋美,李燕.山楂汁澄清工艺的研究[J].饮料工业.2001(2)：32-35.
[61]张卫国,郭红辉,刘欣.明矾澄清灵芝饮料效果的研究[J].广州食品工业科技.2003(3)：27-28.
[62]杨宏图,常翠,饶曼妮,等.五味子宁神口服液澄清工艺改进[J].中国药师.2006(11)：1065-1066.
[63]吴丹,薛旭初,马子骏,等.几种澄清剂在水蜜桃发酵酒中的应用[J].食品工业.2006(4)：11-13.
[64]谭勇,刘四新,柏玉娜,等.几种澄清剂对腰果梨酒品质的影响[J].食品科学.2009(21)：21-23.
[65]张陈云,刘金福,王娜.不同澄清剂对冬枣酒澄清效果的研究[J].食品科技.2010(8)：134-137.
[66]吴国卿,王文平,田亮,等.不同澄清剂对野木瓜果汁澄清效果的影响[J].贵州农业科学.2010(7)：171-172.
[67]陈娟,阚建全,杨蓉生,等.蜂蜜桑椹酒的澄清研究[J].食品工业科技.2010(6)：203-206.
[68]苏槟楠,谢红,刘春卉,等.低糖型金耳饮料口感调配研究初探[J].太原科技.2003(6)：75-76.
[69]刘春凤,郑飞云,李永仙,等.啤酒口感品评的模糊综合评价法[J].食品科学.2008(4)：138-142.
[70]刘春凤,董建军,郑飞云,等.模糊综合评价法在啤酒口感协调性品评中的应用[J].西北农林科技大学学报(自然科学版).2008(3)：213-222.
[71]乔海鸥,丁晓雯,张庆祝,等.柑桔类果汁的脱苦技术[J].包装与食品机械.2003(5)：27-31.
[72]左安连,毛海舫,李琼,等.柑橘类果汁脱苦方法研究综述[J].香料香精化妆品.2008(3)：33-39.
[73]梁泽建.柑桔汁脱苦技术的研究[D].四川农业大学,2007.
[74]潘利华,徐迪,布文婕.柑橘类果汁脱苦的研究进展[J].饮料工业.2006(2)：6-9.
[75]徐国胜,潘利华,杨阳,等.β-环糊精对柑橘类果汁脱苦效果的研究[J].安徽农业科学.2006(20)：5366-5367.