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玛咖活性成分及抗疲劳功能研究
查圣华
学位类型博士
导师赵兵
2016-07
学位授予单位中国科学院研究生院
学位授予地点北京
学位专业生物化工
关键词玛咖 多糖 生物碱 抗疲劳 动物实验
摘要

玛咖(Lepidium meyenii,maca)为十字花科独行菜属草本植物,主要原产地海拔3500-4500米的秘鲁安第斯山区,作为药食两用植物长期食用。我国在云南丽江等地引种驯化,2008年试种成功,2011年被卫生部批准为新资源食品。玛咖的种植规模日益扩大,但云南与原产国秘鲁玛咖的生长环境有所不同,在成分富集上可能存在差异,因此迫切需要相关技术以解决不同产地玛咖品质差异鉴别并解决玛咖多种功能成分集成高效提取、玛咖功效充分利用及安全性与有效性评价等问题。为解决上述问题,本论文对云南产玛咖与秘鲁产玛咖的成分进行了分析比较;对玛咖生物碱与多糖的同步提取进行了研究;并对玛咖多糖的分级制备及其抗氧化性能进行了研究;通过动物实验系统地研究了玛咖及其与中药复合物抗疲劳功效与安全性。取得的主要研究结果如下:(1)云南产玛咖与秘鲁产玛咖成分的比较对云南种植玛咖营养成分、次生代谢产物、矿物质、微量元素以及重金属等指标进行全面分析,并与秘鲁原产地玛咖成分进行比对。结果表明:云南产玛咖与秘鲁产玛咖相比,营养成分如蛋白质(16.32%,12.49%)、腺苷(214.88 μg/g , 25.49 μg/g)、氨基酸(14.68%,11.46%)含量比秘鲁玛咖更高,尤其在与增强生育力及抗疲劳功能相关的成分精氨酸(1.71%,1.17%)、多糖(9.51%,9.42%)、生物碱(0.126%,0.083%)以及玛咖酰胺(0.06%,0.05%)含量上高于秘鲁玛咖,由此可见,云南玛咖在主要功能上有可能优于秘鲁玛咖,可以进行功能研究和深度开发。另外,首次发现玛咖中腺苷含量随时间变化的增长趋势。(2)玛咖生物碱与多糖的同步提取及工业化制备以生物碱和多糖为指标,采用正交试验法研究pH、液固比、提取时间对提取率的影响。各因素对提取结果的影响顺序为:pH>液固比>提取时间,最佳提取工艺条件为提取90 min,提取2次,盐酸调pH 4.5,液固比为10:1;在此条件下,生物碱和多糖的提取率分别达82.91%和85.73%。然后进行了减压、冷冻、喷雾干燥工艺的考察。确定喷雾干燥的条件为:提取液相对密度1.07 g/mL,进风温度165℃,进料流速400 mL/min,干燥过程中生物碱和多糖保留率分别达93.30%和94.10%,提取干燥工艺最终获得放大生产验证,工艺满足产业化生产需要。(3)玛咖多糖的分级制备及其抗氧化活性研究从玛咖水提物中分离玛咖多糖,经Sevage法除蛋白,淀粉酶和糖化酶除淀粉后,通过不同浓度的乙醇溶液分离出4种玛咖多糖(LMP-60、LMP-70、LMP-80和LMP-90)。这些多糖均由鼠李糖、阿拉伯糖、葡萄糖及半乳糖组成,单糖比例分别为1.81:6.85:1:3.21,2.18: 9.47: 1: 5.21,1.49:6.87:1:3.5和0.83: 2.68: 1: 1.32。4种多糖的抗氧化活性与其纯度成正相关,对超氧自由基的清除率(浓度2 mg/mL)均高于玛咖提取物(33.0%),清除率依次为:85.88%、70.15%、60.03%、37.64%。(4)玛咖抗疲劳功能研究将160只健康雄性小鼠随机分为4组,分别进行负重游泳及血清尿素氮(BUN)、肝糖原(HG)和血乳酸(BLA)的测定。结果表明:与阴性对照组相比,低、中、高剂量组小鼠负重游泳时间分别提高了39.46%、66.67%和123.44%,高剂量组BUN与BLA分别降低了17.31%和7.08%,HG三个剂量组平均提高33.35%。表明云南玛咖提取物具有明显的缓解疲劳作用,且有明显的量-效关系。(5)玛咖提取物与淫羊藿提取物复配食用安全性评估进行了玛咖提取物与淫羊藿提取物复配的小鼠急性经口毒性、三项遗传毒性试验和大鼠30 d喂养试验。急性经口毒性试验观察2周后未见动物有中毒症状,属无毒级。遗传毒性试验结果均为阴性。30 d喂养试验:以分别相当于人体推荐量100、50、25倍的3个剂量的样品喂养30 d,与对照组比较,各剂量组的动物体重、增重量、进食量、食物利用率、血常规指标、血生化指标、脏器重量及脏器/体重比值等均无显著差异(P>0.05)。(6)玛咖提取物与淫羊藿提取物复配抗疲劳功能增效作用的评价选用昆明种雄性小鼠,按低、中、高剂量分别给予玛咖提取物与淫羊藿提取物复方、玛咖提取物、淫羊藿提取物及阴性对照组(等量纯水),连续灌胃30 d后,进行抗疲劳指标测定。结果显示:与同等剂量单一提取物相比,复方组能增强小鼠游泳耐力,加速小鼠BUN清除,促进HG储备量,减少运动后BLA增加方面作用显著。

其他摘要

Maca (Lepidium meyenii) is a crucifer plant belonging to the Brassicaceae family. It grows in Andes at altitudes between 3700 m and 4450 m. Since 2008,maca was planted in Lijiang, Yunnan province. It was approved as a new resource food in 2011 in China. The components in Maca from Yunnan and Peru were analyzed systematically in this thesis. Simultaneous extraction of alkaloid and polysaccharide was researched. High-purity maca polysaccharides were obtained by grade preparation and their antioxidant activities were investigated. Finally, anti-fatigue function and safety of maca and its combination with traditional Chinese medicine were evaluated by animal studies. The main results obtained in this paper are as follows:(1) Component analysis of maca from Yunnan and PeruA comprehensive analysis of nutrients, secondary metabolic products, minerals, trace element and heavy metals was conducted. Results showed that maca from Yunnan contained more protein (16.32% to 12.49%), adenosine (214.88 μg/g to 25.49 μg/g) and amino acid (14.68% to 11.46%) than those from Peru, especially the contents of arginine (1.71% to 1.17%), polysaccharide (9.51% to 9.42%), alkaloid (0.126% to 0.083%) and macamide (0.06% to 0.05%), these all related to fertility and anti-fatigue are higher than those from Peru. In addition, the time-variant content of adenosine in maca was found for the first time.(2) The alkaloid and polysaccharide prepared by simultaneous extraction and scaled upThe orthogonal array test was used to determine the optimal extraction conditions, which were as follows: extraction twice, 90 min each time, pH 4.5, solid to liquid 1:10, and the extraction yield of alkaloid and polysaccharide reached 82.91% and 85.73% respectively. Then different drying methods such as vacuum, freeze and spray drying were investigated, and finally the spray drying conditions were as follows: the density 1.07 g/mL, inlet temperature 165℃ and feed rate 400 mL/min. Recovery yield were 93.30% and 94% respectively for alkaloids and polysaccharides.(3) Grading preparation of maca polysaccharides and their antioxidant activitiesThe polysaccharides were deproteinized by Sevage method. Then amylase and glucoamylase were added in order to remove starch. Four kinds of maca polysaccharides (LMP-60、LMP-70、LMP-80 and LMP-90) were obtained by changing the ethanol concentration in the precipitation. All of the LMPs were composed of rhamnose, arabinose, glucose and galactose (1.81:6.85:1:3.21,2.18: 9.47: 1: 5.21,1.49:6.87:1:3.5,0.83: 2.68: 1: 1.32). There was a positive correlation between the purity of LMPs and antioxidant activities. The scavenging rates for superoxide radical were higher than that of maca extract (the rate was 33.0%) with 85.88%、70.15%、60.03%、37.64% for LMP-60、LMP-70、LMP-80 and LMP-90, respectively.(4) Anti-fatigue of maca extractTotal of 160 healthy male mice were randomly divided into 4 groups to examine load swimming time, BUN, HG, and BLA, respectively. Results showed that compared with control group, load swimming time and the content of HG in different dosage groups increased, while the content of urea nitrogen and lactic acid decreased. Therefore, maca extract powder could alleviate physical fatigue.(5) Safety evaluation of maca extract and epimedium extract combinationThe acute toxicity test by oral administration in mice,three terms of genetic toxicity test and the 30 day feeding test in rats were used to evaluate the safety of maca extract and epimedium extract combination. No obvious toxic symptoms and death were found during the 2 week observation.Negative results were observed in the three terms of genetic toxicity test. Dosages of sample, which were corresponding to 100, 50, 25 times daily dosage in human respectively, were fed to the rats for 30 days.Compared with the control group,indexes of weight, weight gain, food intake, food utilization, blood routine index, blood biochemical indicators, viscera weight and viscera /weight ratio in the three dose groups were not significantly different ( P>0.05) .(6) Anti-fatigue of maca extract and epimedium extract combinationMale mice were used to test the anti-fatigue of maca extract and epimedium extract combination, maca extract and epimedium extract with different doses for 30 days. The result showed combination group had better effect than that of the single extract group with the same dosage. The former could prolong the swimming time, accelerate BLA clearance, promote HG storage, and reduce the content of BUN

语种中文
文献类型学位论文
条目标识符http://ir.ipe.ac.cn/handle/122111/22899
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查圣华. 玛咖活性成分及抗疲劳功能研究[D]. 北京. 中国科学院研究生院,2016.
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