摘要
玛咖(Lepidium meyenii Walp.)是生长在秘鲁海拔4000～4500米的安第斯山脉的一种十字花科植物，具有提高生育能力、抗疲劳、改善性功能、抗氧化、抑制前列腺增生、缓解更年期综合症、增强免疫力等多种功效，因此受到世界各国的广泛关注。研究者认为玛咖的多种保健功能与其含有的生物碱、芥子油苷、甾醇、多酚等次生代谢物质有密切关系，而抗氧化活性也是产生这些功效的基础，因此玛咖抗氧化功能及活性物质基础的研究对玛咖其他生物功能机制的揭示、功能产品的开发具有重要的意义。
本论文以云南种植的紫、黄、白3种色型的玛咖为原料，对生物碱、芥子油苷、甾醇、多酚等次生代谢物质的组成及含量进行分析比较，建立和确定了玛咖中次生代谢物质的分析检测方法；通过体外抗氧化活性的检测分析，对抗氧化成分提取及活性进行研究；采用生物活性追踪的方法，筛选出活性较强部分；利用硅胶柱层析的方法将活性较强部位进一步分离为不同的组分，通过各组分次生代谢物质含量和体外抗氧化活性的相关性研究揭示玛咖抗氧化功效的物质基础。并利用对氧化损伤细胞的保护作用进一步验证抗氧化活性组分的抗氧化能力。主要研究结果如下：
1、玛咖次生代谢物质组成与含量分析
（1）总生物碱：通过对玛咖总生物碱含量比色法与滴定法测定结果的比较，确定酸性染料比色法是测定玛咖总生物碱含量较理想的方法，确定了比色测定的方法与条件：测定波长为414nm，溴麝香草酚蓝显色剂的ph值为7.0，用量为5.0mL。比色法的测定结果显示紫色、黄色、白色玛咖的总生物碱含量分别为4.4078、2.2241、2.9193mg/g，3种颜色的差异达到了极显著水平（P＜0.01），其中紫色玛咖的总生物碱含量最大，与相关文献测定的来源于秘鲁的玛咖干粉的总生物碱含量相近。
（2）芥子油苷：采用LC-ESI/MS对玛咖中芥子油苷组成进行的分析和研究表明，3种颜色玛咖中含有相同组分的芥子油苷，即苄基芥子油苷（Benzyl glucosinolate）和甲氧基芐基芥子油苷（Methoxybenzyl glucosinolate），组分中的苄基芥子油苷含量较高，甲氧基芐基芥子油苷含量相对较低，结果与相关文献报道的秘鲁产原料一致。紫、黄、白玛咖鲜样中的芥子油苷总量分别是50.14、46.35、84.57μmol/g，存在显著差异(P＜0.01)，其中白色最高。干燥样品的含量显著低于鲜样（P＜0.01），而不同的干燥方法之间也存在显著差异（P＜0.01），表明不同处理条件对玛咖芥子油苷的水解具有较大影响。
（3）甾醇：采用GC-MS对玛咖的甾醇类化合物组成及含量进行分析，研究表明3种色型玛咖中含有相同组分的甾醇类化合物，即β-谷甾醇（β-Sitosterol）和菜油甾醇（Campesterol）。组分中的β-谷甾醇含量较高，菜油甾醇含量相对较低。白、紫、黄玛咖中的甾醇总量分别是36.60、32.38、27.97mg/100g，存在显著差异（P＜0.01），其中白色含量最高。结果显示甾醇的主要组成与秘鲁产玛咖相同。
（4）多酚：采用Folin-酚测定法对玛咖中的总多酚含量进行分析，结果表明云南种植的紫、黄、白3种颜色玛咖的多酚含量分别为5.08、4.05、3.57mg/g，3种颜色之间存在显著差异（P＜0.01），其中紫色玛咖含量最高。
2、玛咖抗氧化活性成分提取及活性部位研究
采用超氧阴离子自由基的清除能力、羟自由基清除能力、还原抗氧化能力及抑制脂质氧化能力等体外抗氧化评价方法，对甲醇和不同浓度乙醇的玛咖提取物的抗氧化活性进行分析，确定了70%乙醇溶液为玛咖抗氧化活性成分最佳提取溶剂，试验结果也表明了云南种植的紫、黄、白3种颜色玛咖具有明显的体外抗氧化活性，抗氧化活性大小顺序为紫色＞黄色＞白色。活性追踪方法显示黄色和紫色玛咖抗氧化活性较强部位为氯仿极性部位。采用H202诱导损伤细胞保护作用的评价方法对黄色和紫色玛咖氯仿部分及氯仿部分的柱分离组分的抗氧化活性进行验证，结果显示柱分离组分对氧化损伤细胞有明显的保护作用，进一步验证了玛咖的抗氧化活性。
3、玛咖抗氧化活性物质基础的研究
利用相关性分析及多元线性回归的统计方法研究玛咖柱分离组分抗氧化活性与次生代谢物的相关性，研究结果显示，黄色玛咖中多酚含量和生物碱含量与还原抗氧化能力、羟自由基清除能力及脂质氧化抑制率都具有极显著的线性相关（P＜0.01），并且显示抗氧化功效是多酚和生物碱协同作用的结果；紫色玛咖中多酚含量和生物碱含量与还原抗氧化能力呈显著的线性相关（P＜0.01），研究结果揭示了玛咖的主要次生代谢物中多酚和生物碱是玛咖抗氧化能力的主要物质基础。
Maca (Lepidium meyeni) is a plant of the Brassicaceae family growing originally at4000～4500meters altitude in the Andean region of Peru．Maca has attracted attention frommany countries around the world, as it has multiple biological functions such asfertility-enhancing, anti-fatigue, improving sexual function, antioxidant, inhibition prostatehyperplasia, reducing menopause syndrome and increasing immunity function. Researchersrecognized these functions were correlative with maca secondary metabolites includingalkaloids,polyphenol, glucosinolates and sterol, and several functions were also attributable toantioxidant. Therefore studies on antioxidant and active substance basis are important forrevealing reactive mechanism and development of functional products.
In this study, HPLC, LC-ESI/MS and colorimetry have been applied to analyzecompositions and contents of alkaloids,polyphenol, glucosinolates and sterol in three colortypes of maca cultivated in Yunnan, meanwhile the methods analyzing secondary metaboliteshave been established. In addition, the crude extract solvent was confirmed by antioxidantactivity evaluation in vitro. Then the crude extracts of maca were separated into severalfractions of different polarities based on the bioassay guided method. The fractions whichshowed best antioxidative activity were selected to separate into six constituents by silica gelcolumn chromatography. The antioxidative substance basis was revealed by relativity analysisbetween antioxidative activity and the content of secondary metabolites of six constituents.Further, the antioxidative capacities of constituents were confirmed by resisting injury toH_2O_2-induced damage cells. The main results are as follow:
1. Analysis of secondary metabolites compositions and contents in maca
The acidic dye colorimetry is more appropriate for the determination of total alkaloids inmaca cultivated in Yunnan, The conditions of method are: determination wave414nm,dibromothymol-sulfonphthalein dye pH7.0, dye volume5.0mL. The contents of total alkaloids in purple, yellow and white maca were4.4078,2.2241and2.9193mg/g, respectively, thedifference between3color types maca is significant (P＜0.01). The content of total alkaloidsin purple maca is the highest. which is close to the content of total alkaloids in maca grown inPeru.
HPLC and LC-ESI/MS were applied to analyze glucosinolate compositions and contentsof3color types maca cultivated in Yunnan. Three color types of maca contain the samecompositions of glucosinolate, which are benzyl glucosinolate and methoxybenzylglucosinolate.The content of benzyl glucosinolate is higher than that of methoxybenzylglucosinolate.The result was consistent with that of sample cultivated in Peru. The totalglucosinolate contents in purple, yellow and white fresh samples were50.14,46.35, and84.57μmol/g, respectively, with a significant difference (P＜0.01). Among them, the content ofglucosinolate in white sample is highest. The contents of dried samples were much lower thanthat in fresh samples. The contents of glucosinolate in sample processed by two dry techniquewere different significantly(P＜0.01) too, which showed that effect on glucosinolatehydrolyzed of processing condition was obvious.
The compositions and contents of sterols in three color types of maca cultivated in Yunnanwere analyzed by GC-MS．The result showed that all the three types of maca containedβ-sitosterol and campesterol． The content of β-sitosterol was higher than that ofcampesterol．The total sterol contents in white，purple and yellow samples were36.60，32.38，27.97mg/100g，respectively．Among them，the total sterol content in white sample was thehighest．The difference in contents was significant among the three types of sample(P＜0.01)．The composition was consistent with that of sample cultivated in Peru.
The contents of total polyphenol in three color types of macacultivated in Yunnan wereanalyzed by Folin-Ciocalteus method. Result indicated that the total polyphenol contents inpurple, yellow and white were35.08,4.05,3.57mg/g，respectively, with a significantdifference (P＜0.01), the content of polyphneol in purple maca is highest.
2. Study on extraction of antioxidative components and the strongest antioxidativefraction
The antioxidative activities of components extracted by methanol and ethanol withvarious concentration from maca were determined by evaluating ferric reducing antioxidantpower, scavenging capacity of superoxide anion and hydroxyl free radical, inhibition of lipidoxidation. Result showed70%ethanol was appropriate extract solvent for anitoxidativecomponents in maca, The results indicated that ethanol extracts of3color types of macacultivated in Yunnan all had antioxidative activities, among them, the activity of purple macawas highest,and the activity of yellow maca was higher than that of white maca. It was foundthat the chloroform fractions in purple and yellow maca had the strongest antioxidativecapacities by using the bioassay guided method. The antioxidative activities of chloroformfractions and activity constituents separated from chloroform fractions were also confirmed byresisting injury to H_2O_2-induced damage cells, the protective effects of constituents separatedfrom chloroform fractions by silica gel column chromatography were visible in this test.
3. Study on antioxidant substance basis of maca
The relativities were studied between antioxidative activity and the content of secondarymetabolites of constituent separated from chloroform fractions by correlation and linearregression statistic analysis.Result indicated the contents of polyphenol and alkaloids hadsignificant linear correlation with reducing antioxidative capacity, scavenging capacity ofhydroxyl free radical and inhibition of lipid oxidation (P＜0.01) in yellow maca. The resultalso showed that antioxidant capacities resulted from synergic action of polyphenol andalkaloids. In purple maca the contents of polyphenol and alkaloids had significant linearcorrelation with reducingl antioxidative capacity (P＜0.01). The research reveals thatpolyphenol and alkaloids are the most important substance basis for antioxidation. of maca.
引文
曹耀强,谢新华,陈卫军,等.油茶饼中酚类化合物的抗氧化性研究.食品科技,2012,37(3):201-204
陈军,蔡宝昌,沈春云,等.马钱子总生物碱的含量测定.中药新药与临床药理,2007,18(6):468-471
陈阳,张浩,王文君,等.栀子黄多酚含量与抗氧化活性的相关性研究.食品工业科技,2011,32(10):125-128
陈燕青,陈思浩,丁德润.玛咖多糖含量的测定.食品科技,2007,2:231-233
陈奕冰,柳鸿敏,冯艳群,等.葛花多糖对Ｘ射线照射小鼠抗氧化应激能力的影响.中国实验诊断学,2012,16(9):1594-1596
杜广香,浦跃武.玛咖生物碱的分离纯化及抗氧化活性研究.华南理工大学硕士学位论文,2011
冯颖,何钊,徐珑峰,等.云南栽培玛咖的营养成分分析与评价.林业科学研究,2009,22(5):696-700
高路,张公信,高云涛.余甘子多糖提取物抗氧化活性研究.中国农学通报,2011,27(20):133-136
高蒙蒙,孙桂波,斯建勇,等.红车轴草总黄酮对H2O2诱导的血管内皮细胞损伤的保护作用.中国药理学通报.2013,29(2):201-207
高珊,童英,熊晓燕,等.玛咖与西洋参缓解力疲劳作用对比试验研究.实验动物科学与管理,2006,23(4):4-6
郭晶,袁天翊,方莲花,等.天然黄酮对缺氧/复氧损伤乳鼠心肌细胞的保护作用及抗氧化作用比较.中国药理学会补益药药理专业委员会成立大会暨人参及补益药学术研讨会会议论文集,2011:11-12
郭玫.中药成分分析.北京:中国中医药出版社,2006:22-23
郭鑫,杨绍娟,曹霞,等.君子兰总生物碱对肝肾组织功能和抗氧化能力的影响.中国实用医药,2009,4(13):36-37
郭新竹,宁正祥.天然酚类化合物及其保健作用.食品工业,2002,(3):28-29
韩明.青梅果多酚提取及其与抗氧化相关性研究.食品研究与开发,2007,28(6):31-34
何军,祝林,奉建芳.附子总生物碱含量测定方法比较.现代中药研究与实践,2003,17(6):20-22
何钊,冯颖,徐珑峰,等.云南种植玛咖乙醇提取物的体外抗氧化活性分析.食品科学,2010,31(15):39-43
胡雪梅.玛咖和新疆雪莲再生体系的建立及新疆雪莲转昆虫抗冻蛋白基因的研究.新疆大学硕士学位论文,2007
黄雨三.保健食品检验与评价技术规范实施手册.北京:清华同方电子出版社,2003
季宇彬,武晓丹,邹翔.硫代葡萄糖苷的研究.哈尔滨商业大学学报(自然科学版),2005,21(5):550-562
贾之慎,邬建敏,唐孟成.比色法测定Fenton反应产生的羟自由基.生物化学与生物物理进展,1996,23(2):184-186
金文闻.药食两用植物玛咖（Lepidium meyenii）的功效物质研究.华中科技大学博士学位论文,2006
金文闻,王晴芳,李硕,等.新疆产玛咖的挥发油成分研究.食品科学,2009,30(12):241-245
腊贵晓,方萍,李亚娟,等.液相色谱质谱联用分离、鉴定芥蓝中脱硫芥子油苷,浙江大学学报(农业与生命科学版),2008,34(5):557-563
李华,李佩洪,王晓宇,等.抗氧化检测方法的相关性研究.食品与生物技术学报,2008,27(4):6-11
李静,聂继云,李海飞,等. Folin-酚法测定水果及其制品中总多酚含量的条件.果树学报,2008,25(1):126-131
李龙,高荫榆,奚长生,等.混甘酯萃取茶多酚对精炼菜油的抗氧化作用.食品科学,2001,22(11):17-18
李培清.美国特种中药材-玛卡在云南会泽试验种植成功栽培技术.云南农村经济,2007,(4):94-95
李培武,赵永国,张文,等.中国甘蓝型油菜硫苷含量及组份分析.中国农业科学,2005,38(7):1346-1352
丽江格林恒信生物种植有限公司.科学种植玛咖优化高寒山区的产业结构.云南科技管理,2009(1):50
连瑞丽,李宇伟,李存法,等.灵芝菌丝体多糖体外抗氧化活性研究.广东农业科学,2012,(19):93-101
令博,王捷,吴洪斌,等.葡萄皮渣多酚超声波辅助提取工艺响应面法优化及抗氧化活性研究.食品科学,2011,32(18):24-29
刘斌,石任兵,周素蓉.苦参汤有效部位总生物碱含量测定方法研究.北京中医药大学学报,2004,27(2):76-79
刘德胜,刘为忠,颜玲.樱桃叶多糖的抗氧化活性研究.中国生化药物杂志.2012,33(5):571-574
刘海英,仇农学,姚瑞祺,等.我国86种药食两用植物的抗氧化活性及其与总酚的相关性分析.西北农林科技大学学报(自然科学版).2009,37(2):173-180.
刘佳,焦士蓉,唐远谋,等.苦丁茶多酚的提取及抗氧化活性.食品科学,2011,32(14):134-138
刘月萍.黑芥子酶提取分离、性质及固定化研究.浙江工商大学硕士学位论文,2007
庞敏,姜绍通.甾醇氧化特性及其在食品中应用研究进展.食品科学,2010,31(23):434-437
浦跃武,王金全.玛咖多糖的抗氧化性研究.安徽农业科学,2009,37(28):13803-13805
任雪松,于秀林.多元统计分析.北京:中国统计出版社,2012,12
任祎,任贵兴,马挺军,等.燕麦生物碱的提取及其抗氧化活性研究.农业工程学报,2008,24(5):265-269
邵芬娟,闫桂琴,张直峰,等.翅果油树叶片中总生物碱抗氧化活性研究.西北植物学报,2008,28(7):1339-1342
沈蓓,袁冬平,伍城颖,等.芡实提取物对SH-SY5Y神经细胞损伤的保护作用及体外抗氧化活性研究.2013,29(1):39-43
沈维治,邹宇晓,刘凡,等.桑叶总多酚含量与体外抗氧化能力的相关性研究.桑业科学,2012,38(6):1073-1078
石雪萍,吴亮亮,高鹏,等.20种食用辛香料抗氧化性及其与黄酮和多酚的相关性研究.食品科学,2011,32(5):83-86
唐丽华,王登良.苦荞茶的功能研究进展.广东茶业,2011,(6):15-17
涂行浩,张弘,郑华,等.响应面法优化微波钝化玛咖黑芥子酶活.食品科学,2011,32(18):148-153
汪艳群.五味子多糖的分离、结构鉴定及免疫活性研究.沈阳农业大学博士学位论文,2012,102-137
王世博,邱景富,白群华,等.黄芪甲苷对H2O2致PC12细胞氧化应激损伤的保护作用.中国药理学通报.2011,27(11)：1603-1609
王雪飞,张华.多酚类物质生理功能的研究进展.食品研究与开发，2012,33(2):211-214
王振宇,刘瑜,周丽萍.大果沙棘黄酮对糖尿病小鼠血脂与抗氧化水平的影响.食品科学,2010,31(7):297-301
文镜,樊蓉.,植物甾醇和植物甾烷醇降胆固醇的功效和安全性.食品科学,2005,26(8):437-442
吴洪,高平章.11种黄酮类化合物抗氧化活性与结构关系的DFT研究.数理医药学杂志,2009,22(4):434-437
吴宏伟,陈建新,杨洪军,等.丹参成分组合与抗氧化活性相关性分析.中国实验方剂学杂志,2009,15(8):68-71
吴时敏,吴谋成.植物甾醇的研究进展与趋向(Ⅱ).中国油脂,2002,27(3):60-63
邬新荣,王岳飞,张士康等.茶多酚保健功能研究进展与保健食品开发.茶叶科学,2010，30（增刊）:501~505
肖培根,刘勇,肖伟.玛卡－全球瞩目的保健食品.中国食品卫生杂志.2006.18(4):396
谢荣芳,瞿熙.玛卡引种及栽培技术.云南农业科技,2008(4):42-43
姚新生,吴立军,吴继洲,等.天然药物化学.北京:人民卫生出版社,2006,352,441-443
于帅,杜彬,杨越冬,等.植物中多酚的研究进展.河北科技师范学院学报,2011,25(3):69-74
余龙江,金文闻.玛咖(Lepidium meyenii.)干粉的营养成分及抗疲劳作用研究.食品科学,2004,25(2):164-166
余龙江,张永忠,金文闻,等.玛咖醇提取物对小鼠的抗衰老作用.中草药,2006,37(1):81-83
杨晶明,王竹,杨月欣.玛咖(Maca)干品营养成分分析与比较.中国食品卫生杂志,2007,19(3):201-205
尹文清,段少卿,张岩,等.钩藤不同溶剂提取物及总生物碱抗氧化活性研究.广西师范大学学报(自然科学版),2010,28(1):31-34
张超,汪水本,陈奉玲,等.两种方法测定山豆根总生物碱含量的研究.安徽中医学院学报,2002,21(3):51-52
张欣,赵新.几种多酚化合物抗氧化性的不同化学评价及相关性分析.食品科学,2008,29(10):85-89
张晓梅,刘晓芳,高云涛,等.葡萄籽中多酚物质的超声提取及其抗氧化活性研究.安徽农业科学,2012,40(25):12341-12342
张娅,李宝才,项朋志,等.松茸多糖超声提取物抗氧化活性研究.化学与生物工程,2011,28(9):75-82
张永忠.玛咖对更年期综合征及骨质疏松的实验研究.华中科技大学博士学位论文,2006
张永忠,余龙江,万军梅,等.玛咖醇提物对正常小鼠免疫功能的影响.天然产物研究与开发,2007,19:274-276
张永忠,余龙江,敖明章.玛咖醇提取物对去卵巢大鼠内分泌激素及血脂水平的影响.中国新药杂志,2008a,17(24):2112-2121
张永忠,余龙江,金文闻,等.玛咖多糖抗氧化保健作用研究.食品科技,2008b,(5):97-99
张永忠,程华.玛咖提取物的激素样活性实验研究.时珍国医国药,2008,19(8):1874-1875
赵明谋,刘晓丽,崔春,等.余甘子多酚响应面法优化提取及其抗氧化活性研究.食品工业科技,2007,28(6):117-120.
赵奇志,赵毅民.阿朴菲类生物碱生物活性研究进展.天然产物研究与开发,2006,18:316-324
郑海音,赵锦燕,刘艳,等.粗叶悬钩子总生物碱对大鼠非酒精性脂肪肝病的抗氧化作用研究.中国中药杂志,2011,36(17):2383-2387
周达,鲁晓翔,罗成.玫瑰花黄酮对糖尿病小鼠的降血糖作用.食品工业科技,2011,32(2):319-321
周佳,阮征,江波,等.蔬菜抗氧化能力及与酚酸和总黄酮相关性研究.食品与机械,2012,28(3):139-143
周林珠,杨祥良,周井炎,等.多糖抗氧化作用研究进展.中国生化药物杂志,2002,23(4):210-212
周吴萍,李军生,韦媛媛,等.不同采收期广西一点红总生物碱含量分析.食品科学,2009,30(6):151-154
Abeysinghe D C, Li X, Sun C d, et al. Pigment contents and their relationship with hydrophilic andlipophilic antioxidant capacities in seven fruit species. Journal of Fruit Science,2008,25（6）:790～796
Afonso V, Champy R, Mitrovic D, et al. Reactive oxygen spdcies and superoxide dismutases: Role in jointdiseases. Joint Bone Spine,2007,74(4):324-329
Antolovieh M, Prenzler P D, Patsalides E, et al. Methods for testing antioxidant activity. Analyst,2002,12(7):183-198
Bagchi D, Bagghi M, Stohs S J, et a1. Free radicals and grape seed proanthocyanidin extract: importance inhuman health and disease prevention．Toxicology,2000,148;187-197
Battino M, Bullon P, Wilson M, et al. Oxidative injury and inflammatory periodontal diseases: the challengeof antioxidants to free radicals and reactive oxygen species. Critical Reviews in Oral Biology&Medicine,1999,10(4):458-476
Benzie I F, Strain J J. The ferric reducing ability of plasma (FRAP) as a measure of antioxidant power: theFRAP assay. Analytical biochemistry,1996,239(1):70-76
Bogania P, Simonini F, Iriti M, et al. Lepidium meyenii (Maca) does not exert direct androgenic activities.Journal of Ethnopharmacology,2006,104:415-417
Bors W, Michel C. Chemistry of the antioxidant effect of polyphenols. Annals of the New York Academy ofSciences,2002,957:57-69
Brinckmann J, Smith E. Maca culture of the Junín plateau. The Journal of Alternative and ComplementaryMedicine,2004,10(3):426-430
Brooks N A, Wilcox G, Walker K Z, et al. Beneficial effects of Lepidium meyenii (Maca) on psychologicalsymptoms and measures of sexual dysfunction in postmenopausal women are not related to estrogen orandrogen content. Menopause,2008,15(6):1-6
Barzilai A, Yamamoto K. DNA damage responses to oxidative stress. DNA Repair,2004,3(8):1109-1115.
Cassels B K, Asencio M, Conget P, et al. Structure-antioxidative activity relationships in benzyl isoquinolinealkaloids. Pharmacological Research,1995,31:103-107
Choi E H, Kang J I, Cho J Y, et al. Supplementation of standardized lipid-soluble extract from maca(Lepidium meyenii) increases swimming endurance capacity in rats. Journal of Function Foods,2012,4:568-573
Chung F, Rubio J, Carla G, et al. Dose-response effects of Lepidium meyenii (maca) aqueous extract ontesticular function and weight of different organs in adult rats. Journal of Ethnopharmacology,2005,98:143-147
Cicero A F G, Bandieri E, Arletti R. Lepidium meyenii Walp. improves sexual behaviour in male ratsindependently from its action on spontaneous locomotor activity. Journal of Ethnopharmacology,2001,75:225-229
Cicero A F G, Piacente S, Plaza A, et al. Hexanic Maca extract improves rat sexual performance moreeffectively than methanolic and chloroformic Maca extracts. Andrologia,2002,34:177-179
Clément C, Grados D A D, AvμLa B, et al. Influence of colour type and previous cultivation on secondarymetabolites in hypocotyls and leaves of maca (Lepidium meyenii Walpers). Journal of the Science of Foodand Agriculture,2010,90:861–869
Clément C, Witschi U, Kreuzer V. The potential influence of plant-based feed supplements on spermquantity and quality in livestock: A review. Animal Reproduction Science,2012,132:1-10.
Cui B, Zheng B L, He K, et al. Imidazole Alkaloids from Lepidium meyenii. Journal of Natural Product.2003,66:1101-1103
Dash R, Acharya C, Bindu P C. Antioxidant potential of silk protein sericin against hydrogenperoxide-induced oxidative stress in skin fibroblasts. Biochemistry and Molecular Biology Reports,2008,41(3):236-241
Dijk C, Driessen A J, Recourt K. The uncoupling efficiency and affinity of flavonoids for vesicles.Biochem．Pharmacology,2000,60:1593-1600
Dini A, Migliuolo G, Rastrelli L, et al. Chemical composition of Lepidium meyenii. Food Chemistry,1994,49:347-349
Dini I. Tenore G C, Dini A. Glucosinolates from maca (Lepidium meyenii). Biochemical Systematics andEcology,2002,30:1087–1090.
Dording C M, Fisher L, Papakostas G, et al. A Double-Blind, Randomized, Pilot Dose-Finding Study ofMaca Root (L. Meyenii) for the Management of SSRI-Induced Sexual Dysfunction. Neuroscience&Therapeutics,2008,14:182–191
Fang W T, Li H J, Zhou L S. Protective effect of prostaglandin E1on human umbilical vein endothelial cellinjury induced by hydrogen peroxide．Acta Pharmacologica Sinica.2010,31（4）:485-492
Fantel A G, Mackler B, Stamps L D, et al. Reactive oxygen species and dna oxidation in fetal rat tissues. FreeRadical Biology and Medicine,1998,25(1):95-103
Ganzera M, Zhao J P, Muhammad I, et al. Chemical profiling and standardization of lepidium meyenii (maca)by reversed phase high performance liquid chromatography. Chemical&Pharmaceutical Bulletin,2002,50(7):988-991
Gasco M, Aguilar J, Gonzales G F. Effect of chronic treatment with three varieties of Lepidium meyenii(maca) on reproductive parameters and DNA quantification in adult male rats. Andrologia,2007a,39:151-158
Gasco M, Villegas L, Yucra S, et al. Dose–response effect of red maca (Lepidium meyenii) on benignprostatic hyperplasia induced by testosterone enanthate. Phytomedicine,2007b,14:460–464
Gonzales C, Rubio J, Gasco M, et al. Effect of short-term and long-term treatments with three ecotypes ofLepidium meyenii (MACA) on spermatogenesis in rats. Journal of Ethnopharmacology,2006,103:448-454
Gonzales-Casta eda C, Rivera V, Chirinos A L, et al. Photoprotection against the UVB-induced oxidativestress and epidermal damage in mice using leaves of three different varieties of Lepidium meyenii (maca).International Journal of Dermatology,2011,50:928–938
Gonzales G F, Ruiz A, Gonzales C, et al. Effect of Lepidium meyenii(maca) roots on spermatogenesis ofmale rats. Asian Journal of Andrology,2001a,3(3):231-233
Gonzales G F, Cordova A, Gonzales C, et al. Lepidium meyenii(Maca) improvwd semen parameters in adultmen. Asian Journal of Andrology,2001b,3(4):301-303
Gonzales G F, Cordova A, Vega K., et al. Effect of Lepidium meyenii (MACA) on sexual desire and itsabsent relationship with serum testosterone levels in adμLt healthy men. Andrologia,2002,34:367–372
Gonzales G F, Rubio J, Chung A, et al. Effect of alcoholic extract of lepidium meyenii(maca) on testicularfunction in male rats. Asian Journal of Andrology,2003a,5(4):349-352
Gonzales G F, Cordova A, Vega k, et al. Effect of Lepidium meyenii (Maca), a root with aphrodisiac andfertility-enhancing properties, on serum reproductive hormone levels in adult healthy men. Journal ofEndocrinology,2003b,176:163-168
Gonzales G F, Gasco M, Cordova A, et al. Effect of Lepidium meyenii (Maca) on spermatogenesis in malerats acutely exposed to high altitude (4340m).Journal of Endocrinology,2004,180:87-95
Gonzales G F, Miranda S, Nieto J, et al. Red maca (Lepidium meyenii) reduced prostate size in rats.Reproductive Biology and Endocrinology,2005,3:1-16
Gonzales G F, Nieto J, Rubio J, Gascoet M. Effect of black maca (Lepidium meyenii) on one spermatogeniccycle in rats. Andrologia,2006,38:166-172
Gonzales G F, Valerio L G. Medicinal plants from peru: a review of plants as potential agents against cancer.Anti-cancer agents in medicinal chemistry,2006,6:429-444
Gonzales G F, Vasquez V, Rodriguez D, et al. Effect of two different extracts of red maca in male rats withtestosterone-induced prostatic hyperplasia. Asian Journal Andrology,2007,9(2):245–251
Gonzales G F, Gasco M, Malheiros-Pereira A, et al. Antagonistic effect of Lepidium meyenii (red maca) onprostatic hyperplasia in adult mice. Andrologia,2008,40:179–185
Gonzales-Castaneda C, Gonzales G F. Hypocotyls of Lepidium meyenii (maca), a plant of the Peruvianhighlands, prevent ultraviolet A-, B-, and C-induced skin damage in rats. Photodermatology,Photoimmunology&Photomedicine,2008,24:24–31
Harman D. The free radical theory of aging. Antioxid Redox Signal,2003,5(5):557-561
Hatano T, Miyatake H, Natsume M．Proanthocyanidin glycosides and related polyphenols from cacao liquorand their antioxidant effects. Phytochemistry,2002,59:749-758
Hong J, Smith T J, Ho C T．Effects of purified green and black tea polyphenols on cyclooxygenase-andlipoxygenase dependent metabolism of arachidonic acid in human colon mucosa and colon tumor tissues.Biochemical Pharmacology,2001,62:1175-1183
Huang D J, Ou B X, Prior R L. The chemistry behind antioxidant capacity assays. Journal of Agriculturaland Food Chemistry,2005,53:1841-1856
Ikeuchi M, Koyama T, Takei S, et al. Effect Benzylglucosinolate on Endurance Capacity in Mice. Journal ofHealth Science,2009,55(2):178-182
Jin W W, Zhang Y Z, Mei S, et al. Identification of lepidiurm meyenii(walp.) based on spectra andchromatographic characteristics of its principal functional ingredients. Journal of the Science of Food andAgriculture,2007,87:2251-2258
Kiddle G, Bennett R N, Botting N P, et al. High-performance liquid chromatographic separation of naturaland synthetic desμLphoglucosinolates and their chemical validation by UV, NMR and chemicalionisation-MS methods. Phytochemical Analysis,2001,12:226-242
Kim S Y, Kim R H, Huh T L, et al. α-Phenyl-N-t-butylnitrone protects oxidative damage to HepG2cells.Journal of Biochemistry and Molecular Biology,2000,34(1):43-46.
Koufen P, Stark G. Free radical induced inactivation of creatine kinase: sites of interaction, protection, andrecovery. Biochimca et Biophysica Acta,2000,1501,44-50
Kuiper G G, Lemmen J G, Carlsson B, et al. Interaction of estrogenic chemicals and phytoestrogens withestrogen receptor beta. Endocrinol,1998,139:4252-4263．
Lee K J, Dabrowski K, Rinchard J, et al. Supplementation of maca (Lepidium meyenii) tuber meal in dietsimproves growth rate and survival of rainbow trout Oncorhynchus mykiss (Walbaum) alevins andjuveniles. Aquaculture Research,2004,35:215-223
León J. The maca (Lepidium meyenii), a little-known food plant of Peru. Economic Botany,1964,18(2):122-127
Li G Y, Ammermann U, Quiros C E. Glucosinolate contents in maca (Lepidium peruvianum Chacon) seeds,sprouts, mature plants and several derived commercial products. Economic Botany,2001,55(2):255-262
Liu R H, Finley J. Potential cell culture models for antioxidant researeh. Journal of Agricultural and FoodChemistry,2005,53:4311-4314
Lopes G K B, SchμLman H M, Hermes-Lim M．Polyphenol tannic acid inhibits hydroxyl radical formationfrom Fenton reaction by complexing ferrous ions1. Biochimica et Biophysica Acta,1999,1472:142-152
Matés J M, Sánchez-Jiménez F M. Role of reactive oxygen species in apoptosis: implications for cancertherapy. The International Journal of Biochemistry&Cell Biology,2000,32(2):157-170
Matthaus B, Luftmann H. Glucosinolates in members of the family Brassicaceae: separation andidentification by LC/ESI-MS-MS. Journal of Agricultural and Food Chemistry,2000,48(6):2234-2239
Mccollom M M, Villinski J R, Mcphail K L, et al. Analysis of macamides in samples of maca(lepidiummeyenii) by hplc-uv-ms/ms. Phytochemical Analysis,2005,16:463-469
Messina M．A brief historical overview of the past two decades of soy and isoflavone research. Journal ofNutrition,2010,140:1350-1354
Miura T, Motoki H, Naito Y, et al.Antihypoglycemic effect of maca in fasted and insulin-inducedhypoglycemic mice. Journal Traditional Medicines,1999,16:93-96
Moon J K, Shibamoto T. Antioxidant assays for plant and food components. Journal of Agricultural andFood Chemistry,2009,57(5):1655-1666
Muhammad I, Zhao J P, Dunbar D C, et al. Constituents of Lepidium meyenii ‘maca’. Phytochemistr,2002,59:105-110
Nakashima A,Ohtawa M,Masuda N, et al. Antioxidative effects of fluvastatin, and its major metabolites.Yakμgaku Zasshi,2001,121(1):113-116
Niki E. Assessment of antioxidant capacity in vitro and in vivo. Free Radical Biology&Medicine.2010,49(4):503-515.
Orie N N, Zidek W, Tepel M. Reactive oxygen species in essential hypertension and Non–Insulin-Dependentdiabetes mellitus. American Journal of Hypertension,1999,12(12):1169-1174
Oshima M, Gu Y, Tsukada S. Effects of Lepidium meyenii walp and Jatropha macrantha on blood levels ofestradiol-17β, progesterone, testosterone and the rate of embryo implantation in mice. Journal ofVeterinary Medical Science,2003,65(10):1145-1146
Patel R P, Moellering D, Murphy-ullrich J, et al. Cell signaling by reactive nitrogen and oxygen species inatherosclerosis.Free Radical Biology and Medicine,2000,28(12):1780-1794
Piacente S, Carbone V, Plaza A, et al. Investigation of the tuber constituents of maca (Lepidium meyeniiWalp.). Journal of Agricultural Food Chemistry.2002,50:5621-5625
Prior R L, Wu L, Schaich K. Standardized methods for the determination of antioxidant capacity andphenolics in foods and dietary supplements. Journal of Agricultural and Food Chemistry,2005,53(10):4290-4302
Quirós C F, Epperson A, Hu J, et al. Physiological studies and determination of chromosome number inMaca, Lepidium meyenii (Brassicaceae). Economic Botany,1996,50(2):216-223
Ranilla L. G.., Kwon Y, Apostolidis E. et al. Phenolic compounds, antivity and in vitro inhibitory potentialagainst key enzymes relevant for hyperglycemia and hypertension of commonly used medicinal plants,herbs and spices in Latin America.Bioresource Technology,2010,101:4676-4689.
Rea J. Neglected crops:1492from a different perspective//Hernando B, Leon J. Plant production andprotection series No.26. Rome (Italy): FAO,1994:165-167
Reuter S, Gupta S C, Chaturvedi M M, et al. Oxidative stress, inflammation, and cancer: How are theylinked? Free Radical Biology and Medicine,2010,49(11):1603-1616
Riec-Evans C, Mi1ler N J. Structure-antioxidant activity relationships of flavonoidsand phenolic acids. FreeRadical Biology&Medicine,1996,20:933-956
Rondán-Sanabria G G, Finardi-Filho F. Physical-chemical and functional properties of maca root starch.Food Chemistry,2009,114:492-498
Rubio J, Caldas M, Dávila S, et al. Effect of three different cultivars of Lepidium meyenii (maca) on learningand depression in ovariectomized mice. Complementary and Alternative Medicine,2006a,6:23
Rubio J, Riqueros M I, Manuel G, et al. Lepidium meyenii (Maca) reversed the lead acetate induced-Damageon reproductive function in male rats. Food and Chemical Toxicology,2006b,44:1114-1122
Rubio J, Dang H, Gong M, et al. Aqueous and hydroalcoholic extracts of Black maca (Lepidium meyenii)improve scopolamine-induced memory impairment in mice. Food and Chemical Toxicology,2007,45:1882–1890
Rubio J, Yucra S, Gasco M, et al. Dose–response effect of black maca (Lepidium meyenii) in mice withmemory impairment induced by ethanol. Toxicology Mechanisms and Methods,2011,21(8):628–634
Ruiz-Luna A C, Salazar S, Aspajo N, et al. Lepidium meyenii (Maca) increases litter size in normal adultfemale mice. Reproductive Biology and Endocrinology,2005,3:16
Sandoval M. Okuhama N, Angeles F M, et al. Antioxidant activity of the cruciferous vegetable Maca(Lepidium meyenii). Food Chemistry,2002,79:207-213
Han S S, Lo S C, Choi Y W, et al. Antioxidant activity of crude extract and pure compounds of Acer ginnalaMax.. Bulletin of the Korean Chemical Society,2004,25(3):389-391
Stone M, Ibarra A, Roller M. A pilot investigation into the effect of maca supplementation on physicalactivity and sexual desire in sportsmen. Journal of Ethnopharmacology,2009,126(3):574-576
Tellez M R, Khan K A, Kobaisy M, et al. Composition of the essential oil of Lepidium meyenii (Walp.).Phytochemistry,2002,61:149-155
Valentová K, ulrichová J. Smallanthus sonchifolius and Lepidium meyenii-prospective andean crops for theprevention of chronic diseases. Biomedical Papers,2003,147(2):119–130
Valentová K, Buckiova D, Kren V, et al. The in vitro biological activity of Lepidium meyenii extracts. CellBiology and Toxicology,2006,22:91-99
Vecera R, Orolin J, SKottova N, et al. The influence of maca (Lepidium meyenii) on antioxidant status, lipidand glucose metabolism in rat. Plant Foods for Human Nutrition,2007,62:59–63
Wade K L, Garrard I J, Fahey J W. Improved hydrophilic interaction chromatography method for theidentification and quantification of glucosinolates. Journal of Chromatography,2007,1154:469-472
Wang G, Sharma R K, Falcone T, et al. Importance portance of reactive oxygen species in the peritonealfluid of women with endometriosis or idiopathic infertility.Fertility and Sterility,1997,68(5):826-830
Wang L W, Liang J, Wang X D, et al. High efficient antioxidant activity of extracts from Lepidium meyeniiWalp. Asian Journal of Chemistry,2012,24(10):4795-4798
Wang Y, Wang Y, Mcneil B, et al. An Andean crop with mμLti-pharmacological functions. Food ResearchInternational,2007,40:783-792
Wijeratne S S, Cuppett S L, Schlegel V. Hydrogen peroxide induced oxidative stress damage and antioxidantenzyme response in Caco-2human colon cells. Journal of Agricultural and Food Chemistry,2005,53(22):8768-8774
Xu W T, Zhang F F, Luo Y B, et al. Antioxidant activity of a water-soluble polysaccharide purified fromPteridium aquilinum. Carbohydrate Research,2009,344(2):217-22
Yabar E, Pedreschi R, chirinos R, et al. Glucosinolate content and myrosinate activity evolution in threemaca (Lepidium meyenii Walp.)ecotypes during preharvest, harvest and postharvest drying. FoodChemistry,2011,127:1576-1583
Zhang Y Z, Yu L J, Ao M Z, et al. Effect of ethanol extract of Lepidium meyenii Walp. on osteoporosis inovariectomized rat. Journal of Ethnopharmacology,2006,105:274–279
Zhao J, Muhammad I, Dunbar D C, et al. New alkamides from maca (Lepidium meyenii). Journal ofAgricultural and Food Chemistry,2004,53:690–693
Zhao J P, AvμLa B, Chan, M, et al. Metabolomic differentiation of maca(lepidium meyenii) accessionscultivated under different conditions using NMR and chemometric analysis. Planta Medica,2012,78:90–101
Zheng B L, He K, Kim C H, et al. Effect of a lipidic extract from Lepidium meyenii on sexual behavior inmice and rats. Urology,2000,55(4):598-602