本文主要研究内容
作者李媛媛（2019）在《苹果树枝中根皮素的提取纯化、载药体系构建与评价》一文中研究指出：我国苹果种植面积广泛,资源丰富,而每年因苹果树修剪等原因产生大量的苹果树枝,多被废弃,造成资源浪费和环境污染。苹果树枝中含有丰富的黄酮类活性成分,其中二氢查耳酮类化合物根皮苷和根皮素含量较高,根皮素作为根皮苷的苷元发挥主要的药理作用。根皮素具有大多数黄酮类化合物的特性,具有很好的脂溶性,但是水溶性较差,稳定性差,生物利用度低,这些原因使其应用受到了限制。目前针对根皮素的研究主要集中在其药理活性方面,根皮素的提取、纯化及其水溶性改善研究较少,且其提取纯化研究也主要是利用传统萃取分离技术,周期长,得率低,质量不高,而且较难扩大生产。基于上述原因,本研究对苹果树枝中根皮素进行高效绿色提取,分离纯化提取液中的根皮素,获得高纯度根皮素,并以二种天然载体材料对根皮素进行负载,以提高其水溶性和生物利用度。研究结果如下:1、以吐温-80为表面活性剂,纤维素酶和果胶酶为复合酶,采用表面活性剂胶束辅助酶法对苹果树枝进行预处理,提取根皮素的同时将部分根皮苷水解成根皮素,通过单因素和响应面法对预处理方法进行优化,以根皮素总提取率和根皮素提取率为指标(将预处理液中根皮苷按水解反应机理折算成根皮素,计算根皮素提取率),最终得到预处理的最佳条件为:酶解温度为55℃、酶解pH值为4.5、酶解时间为3.5 h、酶的浓度为2.5%、果胶酶与纤维素酶的比例为3:1、表面活性剂的量为5%、料液比为1:11,在此条件下,根皮素总提取率为25.18 mg/g,根皮素提取率为7.26 mg/g。在表面活性剂胶束辅助酶法预处理苹果树枝后,以超声-微波协同法对苹果树枝中的根皮素进行提取,获得最佳提取条件为:超声功率50W,料液比为1:20、表面活性剂的量为6.0%、微波功率为400 W、提取时间为6 min,在此条件下,根皮素总提取率为28.21 mg/g,根皮素提取率为10.51 mg/g。采用表面活性剂胶束辅助酶法预处理,再以超声-微波协同法提取苹果树枝能够增大根皮素的提取率,乙醇为溶剂热回流法根皮素总提取率为22.59 mg/g,根皮素提取率为0.33 mg/g。2、采用表面活性剂胶束浊点分离法将根皮素提取液进行相分离,根皮苷得率为85.4%,根皮素得率为84.9%。氯仿洗涤分相上层液,干燥后可以得到根皮苷含量为8.9%,根皮素含量为3.2%的固体粉末,进一步采用乙醇洗涤去除多糖等杂质、乙醇洗涤液加稀盐酸将根皮苷水解转化为根皮素,稀碱溶液中和,旋转蒸发去除乙醇,乙酸乙酯萃取,最终得到根皮素含量为59.63%的根皮素粗品。以二甲基亚砜为溶剂,水为反溶剂,利用反溶剂沉淀法对根皮素粗品进行纯化,以根皮素的纯度和得率为指标,通过单因素和响应面法优化得到反溶剂沉淀纯化根皮素的最佳件为:溶剂与反溶剂的体积比1:10,沉积时间5 min,沉积温度28℃,根皮素粗品的浓度为46 mg/mL,根皮素纯度和得率分别为98.11%和86.74%。采用高效液相色法、红外光谱法、液相-质谱法和差示量热扫描法对纯化得到的根皮素样品进行测定,确定纯化后得到的样品即为根皮素。3、以多孔淀粉负载根皮素,考察不同因素对多孔淀粉负载根皮素载药量和包封率的影响,确定了多孔淀粉负载根皮素的最佳工艺条件为:吸附时间30 min,根皮素浓度150 mg/mL,根皮素与多孔淀粉的比例1:3,在此条件下,多孔淀粉负载根皮素的载药量为12.8%,包封率为44.4%。通过扫描电镜对根皮素、多孔淀粉、多孔淀粉负载根皮素样品进行形态表征,多孔淀粉负载根皮素后的形态与空白载体基本一致,吸附介质和机械搅拌对多孔淀粉的形态和孔洞没有影响。多孔淀粉吸附根皮素后孔洞被根皮素填充,比表面积明显变小。通过红外光谱、X-射线衍射和差示量热和热重综合分析得出,多孔淀粉负载根皮素后,根皮素的结晶度明显降低,根皮素基本以无定型态存在,热重曲线中,多孔淀粉负载根皮素样品的保留率介于根皮素和多孔淀粉之间,根据各物质的保留率,算出多孔淀粉负载根皮素的载药量为13.7%,结果与HPLC测得的载药量基本一致。4、以乙醇为溶剂介质,将根皮素与羟丙基-β-环糊精以物质的量比1:1制备根皮素包合物,木醋杆菌静态培养得到细菌纤维素,再对细菌纤维素进行冷冻、解冻处理,得到含水率为83.4%的细菌纤维素膜,以细菌纤维素膜对根皮素包合物水溶液进行吸附,冷冻干燥制备细菌纤维素负载根皮素包合物样品,载药量为2.52%,包封率为57.20%。通过扫描电镜、红外光谱、X-射线衍射、差示量热和热重分析表明根皮素与羟丙基-β-环糊精以物质的量1:1形成包合物,细菌纤维素膜中的纤维丝紧密地排列在一起,吸附了根皮素包合物水溶液的细菌纤维在冷冻干燥后呈现出较细菌纤维素膜更加疏松的网状结构,根据热重曲线中各物质的保留率,算出细菌纤维素载药量为2.43%,结果与HPLC测定的载药量结果基本一致。5、测定了根皮素、多孔淀粉负载根皮素、根皮素包合物和细菌纤维素负载根皮素包合物在人工胃液、pH值4.5醋酸-醋酸钠缓冲液和人工肠液中的饱和溶解度,人工胃液介质中分别为 29.07 μg/mL、40.43 μg/mL、49.14 mg/mL 和 48.87 mg/mL,在 pH 值4.5醋酸-醋酸钠缓冲液介质中分别为28.93 μg/mL、44.29μg/mL、59.81 mg/mL、60.12 mg/mL,人工肠液介质中分别为 61.40μg/mL、81.90μg/mL、64.09 mg/mL 和 65.58 mg/mL,载药体系三种介质溶液中的饱和溶解度均明显高于根皮素原药。多孔淀粉负载根皮素、根皮素包合物和细菌纤维素负载根皮素包合物在人工胃液介质中的累积释放率分别是根皮素原粉的12.7倍、17.4倍、9.5倍;在pH值4.5醋酸-醋酸钠缓冲液介质中的累积释放率分别是根皮素原粉的10.4倍、16.2倍、8.7倍;在人工肠液介质中的累积释放率分别是根皮素原粉的7.8倍、7.3倍、5.9倍,载药体系能够明显提高根皮素的溶解度和溶出率。人工胃液、pH值4.5醋酸-醋酸钠缓冲液和人工肠液中的稳定性结果表明,根皮素及其载药体系在人工胃液和pH值4.5醋酸-醋酸钠缓冲液中的稳定性较好,根皮素在人工肠液中较容易发生降解,以多孔淀粉、羟丙基-β-环糊精和细菌纤维素对根皮素负载,能够降低根皮素的降解速度,提高根皮素的稳定性。脂质抗氧化、羟自由基清除能力、还原力测定结果说明根皮素具有很好的体外抗氧化性,而且经过负载后的根皮素体外抗氧化能力优于根皮素原粉。6、对根皮素、多孔淀粉负载根皮素、根皮素包合物和细菌纤维素负载根皮素包合物在大鼠体内血浆药物浓度随时间变化研究表明,多孔淀粉负载根皮素、根皮素包合物和细菌纤维素负载根皮素包合物的生物利用度分别是根皮素原粉的1.89倍、2.39倍、4.56倍。根皮素的组织分布情况表明根皮素、根皮素包合物组大鼠心、肝、脾、肺、脑器官中根皮素最大浓度在给药后的1 h出现,灌胃多孔淀粉负载根皮素、细菌纤维素负载根皮素包合物组在给药2 h后,心、肝、脾、肺、脑器官中根皮素浓度达到最大。多孔淀粉负载根皮素、根皮素包合物、细菌纤维素负载根皮素包合物组大鼠脏器中最大根皮素浓度均高于根皮素原粉组,心脏中,分别是根皮素原粉的1.20、1.21、1.34倍,肝脏中分别是根皮素原粉的1.58、1.88、1.94倍;肺脏中分别是根皮素原粉的1.32、1.76、1.75倍,脾中分别是根皮素原粉的1.63、1.89、1.86倍,脑中分别是根皮素原粉的3.00、3.90、5.10倍;根皮素、多孔淀粉负载根皮素、根皮素包合物、细菌纤维素负载根皮素包合物组大鼠肾脏中根皮素浓度分别在给药后的4 h、6 h、4 h、6 h达到最大值,多孔淀粉负载根皮素、根皮素包合物、细菌纤维素负载根皮素包合物在肾脏中的最大组织浓度分别是根皮素原粉的1.46、1.59、1.83倍。
Abstract
wo guo ping guo chong zhi mian ji an fan ,zi yuan feng fu ,er mei nian yin ping guo shu xiu jian deng yuan yin chan sheng da liang de ping guo shu zhi ,duo bei fei qi ,zao cheng zi yuan lang fei he huan jing wu ran 。ping guo shu zhi zhong han you feng fu de huang tong lei huo xing cheng fen ,ji zhong er qing cha er tong lei hua ge wu gen pi gan he gen pi su han liang jiao gao ,gen pi su zuo wei gen pi gan de gan yuan fa hui zhu yao de yao li zuo yong 。gen pi su ju you da duo shu huang tong lei hua ge wu de te xing ,ju you hen hao de zhi rong xing ,dan shi shui rong xing jiao cha ,wen ding xing cha ,sheng wu li yong du di ,zhe xie yuan yin shi ji ying yong shou dao le xian zhi 。mu qian zhen dui gen pi su de yan jiu zhu yao ji zhong zai ji yao li huo xing fang mian ,gen pi su de di qu 、chun hua ji ji shui rong xing gai shan yan jiu jiao shao ,ju ji di qu chun hua yan jiu ye zhu yao shi li yong chuan tong cui qu fen li ji shu ,zhou ji chang ,de lv di ,zhi liang bu gao ,er ju jiao nan kuo da sheng chan 。ji yu shang shu yuan yin ,ben yan jiu dui ping guo shu zhi zhong gen pi su jin hang gao xiao lu se di qu ,fen li chun hua di qu ye zhong de gen pi su ,huo de gao chun du gen pi su ,bing yi er chong tian ran zai ti cai liao dui gen pi su jin hang fu zai ,yi di gao ji shui rong xing he sheng wu li yong du 。yan jiu jie guo ru xia :1、yi tu wen -80wei biao mian huo xing ji ,qian wei su mei he guo jiao mei wei fu ge mei ,cai yong biao mian huo xing ji jiao shu fu zhu mei fa dui ping guo shu zhi jin hang yu chu li ,di qu gen pi su de tong shi jiang bu fen gen pi gan shui jie cheng gen pi su ,tong guo chan yin su he xiang ying mian fa dui yu chu li fang fa jin hang you hua ,yi gen pi su zong di qu lv he gen pi su di qu lv wei zhi biao (jiang yu chu li ye zhong gen pi gan an shui jie fan ying ji li she suan cheng gen pi su ,ji suan gen pi su di qu lv ),zui zhong de dao yu chu li de zui jia tiao jian wei :mei jie wen du wei 55℃、mei jie pHzhi wei 4.5、mei jie shi jian wei 3.5 h、mei de nong du wei 2.5%、guo jiao mei yu qian wei su mei de bi li wei 3:1、biao mian huo xing ji de liang wei 5%、liao ye bi wei 1:11,zai ci tiao jian xia ,gen pi su zong di qu lv wei 25.18 mg/g,gen pi su di qu lv wei 7.26 mg/g。zai biao mian huo xing ji jiao shu fu zhu mei fa yu chu li ping guo shu zhi hou ,yi chao sheng -wei bo xie tong fa dui ping guo shu zhi zhong de gen pi su jin hang di qu ,huo de zui jia di qu tiao jian wei :chao sheng gong lv 50W,liao ye bi wei 1:20、biao mian huo xing ji de liang wei 6.0%、wei bo gong lv wei 400 W、di qu shi jian wei 6 min,zai ci tiao jian xia ,gen pi su zong di qu lv wei 28.21 mg/g,gen pi su di qu lv wei 10.51 mg/g。cai yong biao mian huo xing ji jiao shu fu zhu mei fa yu chu li ,zai yi chao sheng -wei bo xie tong fa di qu ping guo shu zhi neng gou zeng da gen pi su de di qu lv ,yi chun wei rong ji re hui liu fa gen pi su zong di qu lv wei 22.59 mg/g,gen pi su di qu lv wei 0.33 mg/g。2、cai yong biao mian huo xing ji jiao shu zhuo dian fen li fa jiang gen pi su di qu ye jin hang xiang fen li ,gen pi gan de lv wei 85.4%,gen pi su de lv wei 84.9%。lv fang xi di fen xiang shang ceng ye ,gan zao hou ke yi de dao gen pi gan han liang wei 8.9%,gen pi su han liang wei 3.2%de gu ti fen mo ,jin yi bu cai yong yi chun xi di qu chu duo tang deng za zhi 、yi chun xi di ye jia xi yan suan jiang gen pi gan shui jie zhuai hua wei gen pi su ,xi jian rong ye zhong he ,xuan zhuai zheng fa qu chu yi chun ,yi suan yi zhi cui qu ,zui zhong de dao gen pi su han liang wei 59.63%de gen pi su cu pin 。yi er jia ji ya feng wei rong ji ,shui wei fan rong ji ,li yong fan rong ji chen dian fa dui gen pi su cu pin jin hang chun hua ,yi gen pi su de chun du he de lv wei zhi biao ,tong guo chan yin su he xiang ying mian fa you hua de dao fan rong ji chen dian chun hua gen pi su de zui jia jian wei :rong ji yu fan rong ji de ti ji bi 1:10,chen ji shi jian 5 min,chen ji wen du 28℃,gen pi su cu pin de nong du wei 46 mg/mL,gen pi su chun du he de lv fen bie wei 98.11%he 86.74%。cai yong gao xiao ye xiang se fa 、gong wai guang pu fa 、ye xiang -zhi pu fa he cha shi liang re sao miao fa dui chun hua de dao de gen pi su yang pin jin hang ce ding ,que ding chun hua hou de dao de yang pin ji wei gen pi su 。3、yi duo kong dian fen fu zai gen pi su ,kao cha bu tong yin su dui duo kong dian fen fu zai gen pi su zai yao liang he bao feng lv de ying xiang ,que ding le duo kong dian fen fu zai gen pi su de zui jia gong yi tiao jian wei :xi fu shi jian 30 min,gen pi su nong du 150 mg/mL,gen pi su yu duo kong dian fen de bi li 1:3,zai ci tiao jian xia ,duo kong dian fen fu zai gen pi su de zai yao liang wei 12.8%,bao feng lv wei 44.4%。tong guo sao miao dian jing dui gen pi su 、duo kong dian fen 、duo kong dian fen fu zai gen pi su yang pin jin hang xing tai biao zheng ,duo kong dian fen fu zai gen pi su hou de xing tai yu kong bai zai ti ji ben yi zhi ,xi fu jie zhi he ji xie jiao ban dui duo kong dian fen de xing tai he kong dong mei you ying xiang 。duo kong dian fen xi fu gen pi su hou kong dong bei gen pi su tian chong ,bi biao mian ji ming xian bian xiao 。tong guo gong wai guang pu 、X-she xian yan she he cha shi liang re he re chong zeng ge fen xi de chu ,duo kong dian fen fu zai gen pi su hou ,gen pi su de jie jing du ming xian jiang di ,gen pi su ji ben yi mo ding xing tai cun zai ,re chong qu xian zhong ,duo kong dian fen fu zai gen pi su yang pin de bao liu lv jie yu gen pi su he duo kong dian fen zhi jian ,gen ju ge wu zhi de bao liu lv ,suan chu duo kong dian fen fu zai gen pi su de zai yao liang wei 13.7%,jie guo yu HPLCce de de zai yao liang ji ben yi zhi 。4、yi yi chun wei rong ji jie zhi ,jiang gen pi su yu qiang bing ji -β-huan hu jing yi wu zhi de liang bi 1:1zhi bei gen pi su bao ge wu ,mu cu gan jun jing tai pei yang de dao xi jun qian wei su ,zai dui xi jun qian wei su jin hang leng dong 、jie dong chu li ,de dao han shui lv wei 83.4%de xi jun qian wei su mo ,yi xi jun qian wei su mo dui gen pi su bao ge wu shui rong ye jin hang xi fu ,leng dong gan zao zhi bei xi jun qian wei su fu zai gen pi su bao ge wu yang pin ,zai yao liang wei 2.52%,bao feng lv wei 57.20%。tong guo sao miao dian jing 、gong wai guang pu 、X-she xian yan she 、cha shi liang re he re chong fen xi biao ming gen pi su yu qiang bing ji -β-huan hu jing yi wu zhi de liang 1:1xing cheng bao ge wu ,xi jun qian wei su mo zhong de qian wei si jin mi de pai lie zai yi qi ,xi fu le gen pi su bao ge wu shui rong ye de xi jun qian wei zai leng dong gan zao hou cheng xian chu jiao xi jun qian wei su mo geng jia shu song de wang zhuang jie gou ,gen ju re chong qu xian zhong ge wu zhi de bao liu lv ,suan chu xi jun qian wei su zai yao liang wei 2.43%,jie guo yu HPLCce ding de zai yao liang jie guo ji ben yi zhi 。5、ce ding le gen pi su 、duo kong dian fen fu zai gen pi su 、gen pi su bao ge wu he xi jun qian wei su fu zai gen pi su bao ge wu zai ren gong wei ye 、pHzhi 4.5cu suan -cu suan na huan chong ye he ren gong chang ye zhong de bao he rong jie du ,ren gong wei ye jie zhi zhong fen bie wei 29.07 μg/mL、40.43 μg/mL、49.14 mg/mL he 48.87 mg/mL,zai pH zhi 4.5cu suan -cu suan na huan chong ye jie zhi zhong fen bie wei 28.93 μg/mL、44.29μg/mL、59.81 mg/mL、60.12 mg/mL,ren gong chang ye jie zhi zhong fen bie wei 61.40μg/mL、81.90μg/mL、64.09 mg/mL he 65.58 mg/mL,zai yao ti ji san chong jie zhi rong ye zhong de bao he rong jie du jun ming xian gao yu gen pi su yuan yao 。duo kong dian fen fu zai gen pi su 、gen pi su bao ge wu he xi jun qian wei su fu zai gen pi su bao ge wu zai ren gong wei ye jie zhi zhong de lei ji shi fang lv fen bie shi gen pi su yuan fen de 12.7bei 、17.4bei 、9.5bei ;zai pHzhi 4.5cu suan -cu suan na huan chong ye jie zhi zhong de lei ji shi fang lv fen bie shi gen pi su yuan fen de 10.4bei 、16.2bei 、8.7bei ;zai ren gong chang ye jie zhi zhong de lei ji shi fang lv fen bie shi gen pi su yuan fen de 7.8bei 、7.3bei 、5.9bei ,zai yao ti ji neng gou ming xian di gao gen pi su de rong jie du he rong chu lv 。ren gong wei ye 、pHzhi 4.5cu suan -cu suan na huan chong ye he ren gong chang ye zhong de wen ding xing jie guo biao ming ,gen pi su ji ji zai yao ti ji zai ren gong wei ye he pHzhi 4.5cu suan -cu suan na huan chong ye zhong de wen ding xing jiao hao ,gen pi su zai ren gong chang ye zhong jiao rong yi fa sheng jiang jie ,yi duo kong dian fen 、qiang bing ji -β-huan hu jing he xi jun qian wei su dui gen pi su fu zai ,neng gou jiang di gen pi su de jiang jie su du ,di gao gen pi su de wen ding xing 。zhi zhi kang yang hua 、qiang zi you ji qing chu neng li 、hai yuan li ce ding jie guo shui ming gen pi su ju you hen hao de ti wai kang yang hua xing ,er ju jing guo fu zai hou de gen pi su ti wai kang yang hua neng li you yu gen pi su yuan fen 。6、dui gen pi su 、duo kong dian fen fu zai gen pi su 、gen pi su bao ge wu he xi jun qian wei su fu zai gen pi su bao ge wu zai da shu ti nei xie jiang yao wu nong du sui shi jian bian hua yan jiu biao ming ,duo kong dian fen fu zai gen pi su 、gen pi su bao ge wu he xi jun qian wei su fu zai gen pi su bao ge wu de sheng wu li yong du fen bie shi gen pi su yuan fen de 1.89bei 、2.39bei 、4.56bei 。gen pi su de zu zhi fen bu qing kuang biao ming gen pi su 、gen pi su bao ge wu zu da shu xin 、gan 、pi 、fei 、nao qi guan zhong gen pi su zui da nong du zai gei yao hou de 1 hchu xian ,guan wei duo kong dian fen fu zai gen pi su 、xi jun qian wei su fu zai gen pi su bao ge wu zu zai gei yao 2 hhou ,xin 、gan 、pi 、fei 、nao qi guan zhong gen pi su nong du da dao zui da 。duo kong dian fen fu zai gen pi su 、gen pi su bao ge wu 、xi jun qian wei su fu zai gen pi su bao ge wu zu da shu zang qi zhong zui da gen pi su nong du jun gao yu gen pi su yuan fen zu ,xin zang zhong ,fen bie shi gen pi su yuan fen de 1.20、1.21、1.34bei ,gan zang zhong fen bie shi gen pi su yuan fen de 1.58、1.88、1.94bei ;fei zang zhong fen bie shi gen pi su yuan fen de 1.32、1.76、1.75bei ,pi zhong fen bie shi gen pi su yuan fen de 1.63、1.89、1.86bei ,nao zhong fen bie shi gen pi su yuan fen de 3.00、3.90、5.10bei ;gen pi su 、duo kong dian fen fu zai gen pi su 、gen pi su bao ge wu 、xi jun qian wei su fu zai gen pi su bao ge wu zu da shu shen zang zhong gen pi su nong du fen bie zai gei yao hou de 4 h、6 h、4 h、6 hda dao zui da zhi ,duo kong dian fen fu zai gen pi su 、gen pi su bao ge wu 、xi jun qian wei su fu zai gen pi su bao ge wu zai shen zang zhong de zui da zu zhi nong du fen bie shi gen pi su yuan fen de 1.46、1.59、1.83bei 。
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论文作者分别是来自东北林业大学的李媛媛，发表于刊物东北林业大学2019-11-18论文，是一篇关于苹果树枝论文,根皮素论文,天然载体论文,水溶性论文,生物利用度论文,东北林业大学2019-11-18论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自东北林业大学2019-11-18论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：苹果树枝论文; 根皮素论文; 天然载体论文; 水溶性论文; 生物利用度论文; 东北林业大学2019-11-18论文;