CN106333972B - 黄秋葵抗疲劳有效部位提取物及其制备方法和应用 - Google Patents
黄秋葵抗疲劳有效部位提取物及其制备方法和应用 Download PDFInfo
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- CN106333972B CN106333972B CN201510417143.7A CN201510417143A CN106333972B CN 106333972 B CN106333972 B CN 106333972B CN 201510417143 A CN201510417143 A CN 201510417143A CN 106333972 B CN106333972 B CN 106333972B
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- okra
- antifatigue
- effective part
- extract
- quercetin
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Abstract
本发明公开了一种黄秋葵抗疲劳有效部位提取物及其制备方法和应用。本发明黄秋葵抗疲劳有效部位提取物以总多酚为抗疲劳的主要活性成分,总多酚占有效部位提取物总质量的11.79‑23.34%;所述总多酚主要包括儿茶素类衍生物以及槲皮素衍生物,其中,所述槲皮素衍生物主要包括槲皮素‑3‑O‑龙胆二糖苷、槲皮素‑3‑O‑葡萄糖‑木糖苷、异槲皮苷和槲皮素‑3‑O‑丙二酰基‑葡萄糖苷、芦丁和槲皮素。本发明黄秋葵抗疲劳有效部位提取物能够显著延长小鼠负重游泳时间,消除运动后的乳酸堆积,降低血清尿素氮含量,增加肝糖原,提高小鼠在应激状态下的抗氧化能力,具有极显著的抗疲劳功效。
Description
技术领域
本发明涉及黄秋葵的抗疲劳有效部位提取物及其制备方法,本发明还涉及黄秋葵有效部位提取物在制备抗疲劳的药物、保健品或食品中的用途。
背景技术
疲劳是机体复杂的生理生化变化过程,是人脑力或体力到达一定阶段时必然出现的一种正常的生理现象。它既标志着机体原有工作能力的暂时下降,又可能是机体发展到伤病状态的先兆。据WHO调查,全球约有35%以上的人处于疲劳状态,中年男性人群疲劳状态达60%以上。疲劳的出现,可引起运动能力降低、战斗力减退、工作效率降低、差错事故增多,疲劳发生后如果得不到及时的恢复,逐渐积累,还会导致“过劳”,出现“过度训练综合症”、“慢性疲劳综合症”等,使机体发生内分泌紊乱、免疫力下降,甚至出现器质性病变,成为威胁人类健康的重要因素。
然而,目前市场上抗疲劳的化学药物如苯丙胺类、莫达非尼、咖啡因等在延缓疲劳发生和消除疲劳方面确有较好的效果,但是出现了许多不良反应,如心血管的不良影响、干扰睡眠、精神紊乱、成瘾性等。因此,从天然植物资源中提取安全、有效,不含对人体有害的兴奋剂成分的抗疲劳成分已成为近年来抗疲劳的研究热点。
黄秋葵(Okra)属于锦葵科(Malvaceae)秋葵属(Abelmoschus)植物Abelmoschusesculentus(linn.),别名秋葵、羊角豆,广泛分布于海南、广东、广西、浙江、湖南、四川、河南等省。黄秋葵含有蛋白质、脂肪、多糖、维生素及微量元素等营养成分[高振茂,高冠亚,杜丽红。天然佳蔬黄秋葵的营养与食用方法。上海蔬菜,2005(2):76-77;黄阿根,陈学好,高云中等.黄 秋葵的成分测定与分析。食品科学,2007(10):451-455]以及黄酮等次生代谢产物。已从黄秋葵中分离得到的黄酮主要有:4,5,7-三羟基黄酮、4,5,7-三羟基黄酮-3-散O-葡萄糖苷、杨梅黄素、杨梅黄素-3-O-葡萄糖苷、杨梅黄素-3-O-鼠李糖苷、异鼠李糖-3-O-葡萄糖-戊糖苷、槲皮素、槲皮素-3-O-芸香糖苷、槲皮素-3-O-葡萄糖苷、槲皮素-3-O-二葡萄糖苷、槲皮素-3-O-葡萄糖-木糖苷和槲皮素-3-O-(丙二酰基)葡萄糖苷[ArapitsasP.Identification and quantification of polyphenolic compounds from okra seedsand skins[J].Food Chemistry,2008,110(4):1041-1045;Shui GH,Peng LL.An improvedmethod for the analysis of major antioxidants of Hibiscus esculentus Linn[J].Journal of Chromatography A.2004,1048(1):17-24]。
黄秋葵具有抗肿瘤[任丹丹,陈谷。黄秋葵多糖组分对人体肿瘤细胞的增殖抑制作用。食品科学,2010,31(21):353-356;金忠浇、林建龙。治疗皮肤癌的苗头中草药--秋葵。中华实用中西医杂志。2001,29(10):2256]、抗氧化[Adelakun OE,Oyelade OJ,Omowaye BO,et al.Chemical composition and the antioxidative properties of Nigerian OkraSeed(Abelmoschus esculentus Moench)Flour.Food and Chemical Toxicology,2009,47(6):1123-1126;Adelakun OE,Oyelade OJ,Omowaye BO,et al.Influence of pre-treatment on yield chemical and antioxidant properties of a Nigerian okraseed(Abelmoschus esculentus moench)flour.Food and Chemical Toxicology,2009,47(3):657-661]、肝保护[A study of its hepatoprotective activity[J].SaudiPharmaceutical Journal.2011 Oct 2]、神经细胞保护[Walaiporn T,Nootchanart R,Piyanete C,et al.Neuroprotective effects of quercetin,rutin and okra{Abelmoschus esculentus Linn.)in dexamethasone-treated mice.NeurochemistryInternational,2011,59(5):677-685}]等活性。
尽管黄秋葵抗疲劳的功效已有文献报道,迄今为止,黄秋葵的抗疲劳活性部位及活性组分尚不清楚,严重制约了其在制备药物、保健品或食品中的推广应用。
发明内容
本发明的目的之一是提供一种黄秋葵抗疲劳有效部位提取物;
本发明的目的之二是提供一种制备所述黄秋葵抗疲劳有效部位提取物的方法;
本发明的目的之三是将所述的黄秋葵有效部位提取物应用于制备抗疲劳的药物、保健品或食品。
本发明的上述目的是通过以下技术方案来实现的:
一种黄秋葵抗疲劳有效部位提取物,以总多酚为抗疲劳的主要活性成分,总多酚占有效部位提取物总质量的11.79-23.34%;优选的,总多酚占有效部位提取物总质量的23.34%。
其中,所述的总多酚主要包括儿茶素类衍生物和槲皮素衍生物,所述儿茶素类衍生物主要包括:表没食子儿茶素二聚体、表没食子儿茶素、芥子酰基阿魏酸酯等;所述的槲皮素衍生物:槲皮素-3-O-龙胆二糖苷、槲皮素-3-O-葡萄糖-木糖苷、异槲皮苷和槲皮素-3-O-丙二酰基-葡萄糖苷、芦丁和槲皮素等;其中,槲皮素-3-O-龙胆二糖苷占提取物总质量的1.97-2.99%,异槲皮苷占提取物总质量1.15-2.16%。
本发明的另外一个目的是提供一种制备所述黄秋葵抗疲劳有效部位提取物的方法,该方法包括以下步骤:
(1)鲜用或干用的黄秋葵果实加溶剂回流提取,收集提取液;(2)将所述提取液浓缩后过大孔树脂吸附,弃去水洗脱液,再用乙醇洗脱,收集乙醇洗脱液,浓缩,即得。
所述鲜用的黄秋葵果实是刚收获时嫩果或在冰冻条件下保存解冻后的嫩果;所述干用的黄秋葵果实是指将黄秋葵嫩果干燥后的黄秋葵果实;所述的干燥方式包括:冷冻干燥,阴干或低温烘干中任何一种常规的干燥方式;所述黄秋葵嫩果为坐果4-7天采摘的果实。
步骤(1)中所述的溶剂包括水、30%乙醇或50%乙醇;
步骤(1)中所述回流提取的次数至少进行二次,优选为三次;每次提取时间30-60分钟,优选60分钟;每次提取时,所述黄秋葵与提取溶剂的质量比为1:8-12,优选为1:10。
步骤(2)中所述的浓缩是将提取液浓缩至1/3体积;
步骤(2)中所述的大孔树脂选自D101,DM130,HPD417,HPD750或ADS-17大孔树脂中的任何一种,优选为HPD50树脂;
步骤(2)中所述的乙醇洗脱优选用90%乙醇进行洗脱。
本发明黄秋葵抗疲劳有效部位提取物采用小鼠负重3%铅皮一次性力竭游泳模型,以行为学检测和组织中各类生化指标为参数对黄秋葵抗疲劳有效部位提取物的抗疲劳活性进行研究;实验结果表明,本发明黄秋葵抗疲劳有效部位提取物能够延长小鼠负重游泳时间,消除运动后的乳酸堆积,降低血清尿素氮含量,提高小鼠在应激状态下的抗氧化能力,对疲劳恢复、抗疲劳能力提高具有促进作用;
本发明的另一个目的是将黄秋葵抗疲劳有效部位提取物组合物应用于制备抗疲劳药物、抗疲劳保健品、抗疲劳食品等各种产品。
因此,本发明提供了一种抗疲劳药物组合物,该药物组合物中含有有效量的黄秋葵抗疲劳有效部位提取物以及药学上可接受的载体。
在上述药物中,还可以加入一种或多种药学上可接收的载体或辅料。所述的载体或辅料是指药学领域常规的载体或辅料,例如:稀释剂、崩裂剂、润滑剂、赋形剂、粘合剂、助流剂、填充剂、表面活性剂等;另外,还可以在组合物中加入其它辅助剂如香味剂和甜味剂。
所述稀释剂可以是一种或几种增加片剂重量和体积的成分;常用的稀释剂包括乳糖、淀粉、预胶化淀粉、微晶纤维素、山梨醇、甘露醇以及无机钙盐等。其中最常用为乳糖、淀粉、微晶纤维素。
所述崩解剂可以为交联聚乙烯吡咯烷酮(与总重量比为2-6%),交联羧甲基纤维素钠(与总重量比为2-6%)、海藻酸(与总重量比为2 -5%)、微晶纤维素(与总重量比为5-15%)中之一种或几种混合物。其中以交联聚乙烯吡咯烷酮(与总重量比为2-7%),交联羧甲基纤维素钠(与总重量比为2-6%)为佳。最佳为交联聚乙烯吡咯烷酮(与总重量比为2-6%)。
所述的润滑剂包括硬脂酸,硬脂酸钠,硬脂酸镁,硬脂酸钙,聚乙二醇,滑石粉,氢化植物油中之一种或几种混合物。其中以硬脂酸镁最为适宜。润滑剂的用量范围(与总重量比)为0.10-1%,一般用量为0.25-0.75%,最佳用量为0.5-0.7%。
所述的粘合剂可以是一种或几种有利于制粒的成分。可以是淀粉浆(10-30%,与粘合剂总重量比),羟丙基甲基纤维素(2-5%,与粘合剂总重量比),聚乙烯吡咯烷酮(2-20%,与粘合剂总重量比),以聚乙烯吡咯烷酮的乙醇水溶液为佳,最佳为聚乙烯吡咯烷酮的50%乙醇水溶液。
所述助流剂可以为微粉硅胶、滑石粉、三硅酸镁中之一种或几种混合物。
所述表面活性剂可以为一种或几种能够提高润湿性和增加药物溶出的成分。常用为十二烷基硫酸钠(常用范围为0.2-6%,与总重量比)。
用黄秋葵抗疲劳有效部位提取物制备的抗疲劳药物可以制成注射剂,片剂,粉剂,颗粒剂,胶囊,口服液等多种形式。上述各种类型的药物均可以按照药学领域的常规方法制备。
本发明还提供了一种抗疲劳的保健品,所述保健品中包含本发明提供的黄秋葵抗疲劳有效部位提取物;
此外,本发明的黄秋葵组合物还可进一步将其应用于制备成抗疲劳的食品。
附图说明
图1没食子酸的标准曲线图。
图2黄秋葵多酚提取物的UPLC-QTof离子流图;主要化合物:保留时间tR(min)=4.1:表没食子儿茶素二聚体;tR(min)=7.02:表没食子儿茶素;tR(min)=8.29:芥子酰基阿魏酸酯;tR(min)=9.06:槲皮素-3-O-龙胆二糖苷;tR(min)=9.40:槲皮素-3-O-葡萄糖-木糖;tR(min)=10.29:芦丁;tR(min)=10.79:异槲皮苷;tR(min)=11.88:槲皮素-3-O-(丙二酰基)葡萄糖苷;tR(min)=18.85:槲皮素。
图3黄秋葵有效部位提取物的HPLC色谱图。
具体实施方式
下面结合具体实施例来进一步描述本发明,本发明的优点和特点将会随着描述而更为清楚。但是应理解所述实施例仅是范例性的,不对本发明的范围构成任何限制。本领域技术人员应该理解的是,在不偏离本发明的精神和范围下可以对本发明技术方案的细节和形式进行修改或替换,但这些修改或替换均落入本发明的保护范围。
具体实施方式
下面通过具体实施例对本发明进行说明,但本发明并不局限于此。下述实施例中所述的方法,如无特殊说明,均为常规方法,所述试剂和生物材料如无特殊说明,均可从商业途径获得。
本发明中所用到的对照品没食子酸购自中国药品生物制品检定所,批号分别为110831-201204。槲皮素-3-O-龙胆二糖苷和异槲皮苷购自成都普菲德生物技术有限公司。
实施例1黄秋葵有效部位提取物的制备
1称取黄秋葵嫩果,称取新鲜嫩果2000g(相当于干果200g),加蒸馏水2000ml,回流提取60分钟;
2倾出提取液,药渣按照第一次提取条件和方法重复回流提取二次,合并三次提取液;
3将提取液减压浓缩至2000ml时,采用D101树脂(河北沧州宝恩化工有限公司购买)吸附,用蒸馏水洗脱,收集蒸馏水洗脱液;再用90%乙醇洗脱,收集乙醇洗脱液;
4将蒸馏水洗脱液浓缩、干燥,得到黄秋葵水洗脱提取物(HQKS1);将乙醇洗脱液浓缩、干燥,得到抗疲劳有效部位提取物(HQK1)。
实施例2黄秋葵有效部位提取物的制备
黄秋葵水提取液采用DM130大孔树脂(河北沧州宝恩化工有限公司购买)吸附,其它方法同实施例1完全相同,得到黄秋葵水洗脱提取物(HQKS2)和抗疲劳有效部位提取物(HQK2)。
实施例3黄秋葵有效部位提取物的制备
黄秋葵水提取液采用HPD417大孔树脂(河北沧州宝恩化工有限公司购买)吸附,其它方法同实施例1完全相同,得到黄秋葵水洗脱提取物(HQKS3)和抗疲劳有效部位提取物(HQK3)。
实施例4黄秋葵有效部位提取物的制备
黄秋葵水提取液采用HPD750大孔树脂(河北沧州宝恩化工有限公司购买)吸附,其它方法同实施例1完全相同,得到黄秋葵水洗脱提取物(HQKS4)和抗疲劳有效部位提取物(HQK4)。
实施例5黄秋葵有效部位提取物的制备
黄秋葵水提取液采用ADS-17大孔树脂(河北沧州宝恩化工有限公司购买)吸附,其它方法同实施例1完全相同,得到黄秋葵水洗脱提取物 (HQKS5)和抗疲劳有效部位提取物(HQK5)。
试验例1黄秋葵乙醇洗脱提取物中总多酚含量测定、主要化学成分鉴定及主要化学成分含量的测定
采用紫外分光光度法测定黄秋葵乙醇洗脱液(即:实施例1-5所制备的抗疲劳有效部位提取物)中总多酚含量,采用UPLC-MS鉴定黄秋葵乙醇洗脱液(即:实施例1-5所制备的抗疲劳有效部位提取物)中主要成分,采用高效液相色谱法测定黄秋葵乙醇洗脱液(即:实施例1-5所制备的抗疲劳有效部位提取物)中槲皮素-3-O-龙胆二糖苷和异槲皮苷含量;具体方法如下:
一、紫外分光光度法测定黄秋葵提取物中总多酚含量测定
(一)测定方法
1)标准曲线的绘制
称取没食子酸标准品7mg溶于25mL蒸馏水中,准确吸取0,0.2,0.4,0.6,0.8,1mL于25mL容量瓶中,分别加入FC试剂2mL,10%Na2CO3溶液10mL,蒸馏水定容至25mL,置于50℃水浴中避光反应1h,在765nm处测定吸光度,绘制标准曲线。
2)样品测定
精密称取样品适量,定容于25mL容量瓶中,分别加入FC试剂2mL,10%Na2CO3溶液10mL,蒸馏水定容至25mL,置于50℃水浴中避光反应1h,在765nm处测定吸光度,根据标准曲线计算总多酚的含量。
3)对照设置
在标准曲线的制作和样品测定时,设置对照溶液,于25mL容量瓶中,分别加入FC试剂2mL,10%Na2CO3溶液10mL,蒸馏水定容至25mL,置于50℃水浴中避光反应1h,在765nm处测定吸光度。
(二)总多酚测定结果
通过紫外分光光度法,以吸光值为纵坐标,没食子酸质量浓度(mg/L)为横坐标绘制校准曲线,没食子酸在0.056-0.28mg/ml内具有良好的线性关系。得到没食子酸的回归方程:y=109.13x+0.0054(R2=0.9996)(图1)
分别测定实施例1-实施例5制备的黄秋葵抗疲劳有效部位提取物HQK1-HQK5中总多酚占有效部位提取物的质量,测定结果分别为:HQK1:13.21%,HQK2:12.56%,HQK3:12.72%,HQK4:23.34%,HQK5:11.79%。
测定实施例1-实施例5制备的五个水洗脱部位HQKS1-HQKS5的总多酚含量,总多酚占水洗脱提取物质量分别为:HQKS1:0.11%,HQKS2:0.25%,HQKS3:0.15%,HQKS4:0.05%,HQKS5:0.13%.
二、超高效液相色谱与质谱联用技术(UPLC-ESI-QTof)鉴定黄秋葵有效部位提取物中主要化学成分
(一)鉴定方法
1)超高效液相条件
色谱柱:Acquity UPLC BEH C18(2.1mm×150mm,1.7μm);保护柱:VanGuand BEHC18(2.1mm×5mm,1.7μm),流动相:A为乙腈,B为0.01%甲酸水溶液,梯度洗脱(0→30min:5%→100%A;30→35min:100%A;35→35.01min:100%→5%A;35.01→37min:5%A);流速:0.4mL/min;进样量:1μL;柱温40℃,检测器:PDA:200-400nm全扫描。
2)质谱条件
正离子,MSe模式;检测范围100-1500Da;毛细管电压:3kV,锥孔电压:50V,裂解电压:40-50V;锥孔气流量:50L/h;脱溶剂气流量:800L/h;源温:100℃;脱溶剂气温度350℃;准确质量测定采用亮氨酸-脑啡肽(Leucine-Enkephalin,m/z=556.2771)溶液作为质量锁定溶液。
3)样品测试
将供试品溶液1μL注入液相-质谱系统分析检测,运行时间为30min。每个样品重复提取3次,至少平行进样2次。
4)数据分析
色谱及质谱数据使用沃特世工作站MassLynx4.1软件进行处理。
(二)鉴定结果
通过UPLC-QTof技术,从黄秋葵抗有效部位提取物(HQK1-HQK5)中鉴定出的主要化合物有:儿茶素类衍生物主要包括:表没食子儿茶素二聚体、表没食子儿茶素、芥子酰基阿魏酸酯等;槲皮素衍生物主要包括:槲皮素-3-O-龙胆二糖苷、槲皮素-3-O-葡萄糖-木糖苷、异槲皮苷和槲皮素-3-O-丙二酰基-葡萄糖苷、芦丁和槲皮素等。详见黄秋葵多酚UPLC-QTof主要化合物鉴定色谱图(图2)。
三、HPLC测定提取物中槲皮素-3-O-龙胆二糖苷和异槲皮苷的含量
(一)测定方法
1)色谱条件:Thermo Syncronis AQ-C18色谱柱(4.6mm×250mm,5μm),流动相A为含0.1%甲酸的水溶液,B为乙腈,梯度洗脱,洗脱程序如表1;流速1ml/min;柱温30℃,检测波长:280nm和350nm双波长检测;进样体积10μl,分析时间40分钟。
表1梯度洗脱程序表
时间(min) | 流动相A(1%甲酸水溶液) | 流动相B(乙腈) |
0 | 97 | 3 |
5 | 85 | 15 |
12 | 82 | 18 |
20 | 70 | 30 |
25 | 5 | 95 |
30 | 97 | 3 |
40 | 97 | 3 |
2)标准品溶液的配制
精密称取各标准品5mg,用甲醇溶解并定容于10ml容量瓶,配成0.5mg/ml的标准品溶液,置冰箱中保存备用。
3)样本制备
精密称取实施例1-5所制备的黄秋葵抗疲劳有效部位提取物HQK1-HQK5各200mg,加甲醇10ml,充分溶解后,分别定容于10ml容量瓶。
4)实验步骤
准确吸取标准品溶液和黄秋葵有效部位提取物溶液(HQK1-HQK5)各10μl,按照色谱条件分别进行测定,标识相应色谱峰,计算峰面积。
5)数据分析
Waters色谱工作站Empower软件进行分析。
(二)测定结果
通过高效液相色谱对样品及两个标准品的检测可见色谱图(图3),黄秋葵有效部位提取物(HQK1-HQK5)中槲皮素-3-O-龙胆二糖苷占提取物总质量在1.97-2.99%,异槲皮苷占提取物总质量在1.15-2.16%。
实施例2黄秋葵不同树脂制备有效部位提取物(HQK1-HQK5)及非有效部位(HQKS1和HQKS2)的抗疲劳活性比较试验
1.试验药物
实施例1-5制备的黄秋葵抗疲劳有效部位提取物(HQK1-HQK5)和非有效部位(HQKS1-HQKS5)。
2动物分组及给药
7周龄雄性ICR小鼠(20±1g)100只(北京维通利华实验动物技术有限公司),实验动物合格证号:SCXK(京)2012-0001。100只小鼠随 机分为10组,每组10只。人参剂量:100mg/kg,黄秋葵有效部位提取物HQK1—HQK5剂量:200mg/kg;黄秋葵水洗脱提取物:400mg/kg。
A组:空白对照,灌胃生理盐水;
B组:人参提取物组(阳性对照);
C组:黄秋葵有效部位提取物(HQK1);
D组:黄秋葵有效部位提取物(HQK2);
E组:黄秋葵有效部位提取物(HQK3);
F组:黄秋葵有效部位提取物(HQK4);
G组:黄秋葵有效部位提取物(HQK5);
H组:黄秋葵非有效部位提取物(HQKS1);
I组:黄秋葵非有效部位提取物(HQKS2);
J组:黄秋葵非有效部位提取物(HQKS3);
K组:黄秋葵非有效部位提取物(HQKS4);
M组:黄秋葵非有效部位提取物(HQKS5)。
3实验方法
3.1小鼠负重游泳实验
检测容器:采用直径43cm,高40cm的水桶,水位高25cm,水温25±1℃,4只/桶;检测标准:首次下沉标准为小鼠鼻尖首次没入水面以下,力竭标准为小鼠沉入水下10s不上浮;检测方法:给药30分钟后,将小鼠尾部负上相应体重3%的铅皮,各组随机抽取1只小鼠,放入水中开始计时,到小鼠力竭结束计时,记录小鼠首次下沉时间与力竭时间,两者时间之差表征小鼠的耐力。
3.2动物取材
小鼠力竭游泳检测结束10分钟后,立即对空白组、人参组和黄秋葵 提取物组的小鼠摘取眼球取血,解剖取肝脏组织。血液置4℃冰箱静置一夜,第二天3000r/min,10min离心,取血清;肝脏组织置于﹣80℃保存备用。
3.3生化指标的测定
血清用于测定血乳酸和尿素氮,肝脏用于测定肝糖原、丙二醛(MDA)、超氧化物歧化酶(SOD)和谷光甘肽还原酶(GSH-PX)。所有的生化试剂盒均由南京建成生物工程研究所提供。测定方法根据试剂盒说明书进行。
4统计方法
采用SPSS统计分析软件进行数据处理,所有数据均以平均值±标准误表示,组间差异用单因素方差分析(One-way ANOVA)处理。
5实验结果
5.1小鼠负重游泳力竭时间的测定
从表2的数据可见,五种不同树脂吸附黄秋葵水提取液后,得到黄秋葵多酚提取物,具有明显的抗疲劳效果;但五种不同树脂处理得到多酚提取物,存在显著的差异;其中,HQK4提取物(采取HPD750大孔树脂作为吸附剂制备的黄秋葵提取物)的抗疲劳效果最好。同时,还发现黄秋葵水提取液大孔树脂吸附的水洗脱部位(非多酚部位:HQKS1-HQKS5)基本上没有抗疲劳活性。
表2黄秋葵不同提取物抗疲劳检测结果
*表示P≤0.05,与空白组比较具有统计学差异;
**表示P≤0.01,与空白组比较统计学差异显著;
***表示P≤0.001,与空白组比较统计学差异极为显著。
5.2小鼠负重游泳抗疲劳生化指标的测定
对空白组人参组及HQK1-HQK5组小鼠的血清乳酸(BLA)、尿素氮(BUN)、肝糖原及丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱肝肽还原酶(GSH-PX)进行了测定,结果见表3和表4。
从表3和表4可以看出,黄秋葵提取物HQK1--HQK5五个组:血清乳酸、尿素氮及MDA水平显著低于正常对照组,而超氧化物歧化酶(SOD)、谷胱肝肽还原酶(GSH-PX)及肝糖原水平显著高于对照组。其中,HQK4血清乳酸、尿素氮及MDA水平极显著低于正常对照组,而超氧化物歧化酶(SOD)、谷胱肝肽还原酶(GSH-PX)及肝糖原水平极显著高于对照组;说明从生化指标角度也证明本发明制备的黄秋葵有效部位提取物具有明显的抗疲劳活性。
表3黄秋葵提取物抗疲劳生化指标的检测结果
表4黄秋葵提取物抗疲劳生化指标的检测结果
Claims (3)
1.一种黄秋葵抗疲劳有效部位提取物,其特征在于:以总多酚为抗疲劳的主要活性成分;其中,总多酚占有效部位提取物总质量的23.34%;其制备法包括以下步骤:(1)黄秋葵嫩果加水回流提取3次,每次提取时间为60分钟;按照g:ml计,每次提取时,黄秋葵嫩果与提取溶剂水的比例为1:1;收集提取液;(2)将所述提取液浓缩后用大孔树脂HPD750吸附,用水洗脱,弃去水洗脱液;再用90%乙醇洗脱,收集90%乙醇洗脱液,将乙醇洗脱液浓缩、干燥,即得。
2.按照权利要求1所述的黄秋葵抗疲劳有效部位提取物,其特征在于:所述的干燥方式包括:冷冻干燥,阴干或低温烘干;所述黄秋葵嫩果为坐果4-7天采摘的果实。
3.权利要求1所述的黄秋葵抗疲劳有效部位提取物在制备抗疲劳的药物、保健品或食品中的用途。
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