CN114848692A - 一种藜麦苗多酚类物质的提取工艺及应用 - Google Patents
一种藜麦苗多酚类物质的提取工艺及应用 Download PDFInfo
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Abstract
本发明公开了一种藜麦苗多酚类物质的提取工艺及应用,属于天然产物提取领域。主要包括以下步骤:(1)将藜麦苗烘干粉碎,过100目筛;(2)称取藜麦苗粉末,加入一定量蒸馏水中,‑25℃冷冻2h,60℃水浴融解10min,循环冻融4‑6次;(3)加入无水乙醇使乙醇水溶液浓度为60%‑80%,料液比为1:20;(4)在50℃‑70℃下,300W功率超声提取40‑60min,离心;(5)将上清液旋转蒸发、冷冻干燥成粉末。采用冻融结合超声波辅助提取,可以促进藜麦苗细胞内容物溶出,增加提取率。本发明具有工艺简单、成本低、提取率高、污染小等优点。藜麦苗多酚作为抗氧化剂的有效成分可用于医药、食品及保健品领域。
Description
技术领域
本发明涉及天然产物提取技术领域,具体涉及一种藜麦苗中多酚类物质的提取工艺及应用。
背景技术
藜麦(Chenopodium quinoa Willd.)是藜科藜属的一年生双子叶植物,原产于南美洲印第安山脉,由于其丰富的营养价值和耐干旱耐盐碱特性,被印第安人称为“粮食之母”。作为粮食作物,藜麦主要以收获籽粒为主。近年来,藜麦的幼苗作为一种营养素食,因其丰富的营养价值、较高的保健功能和独特食用风味,逐渐成为餐桌上常见的食物。因此,藜麦苗也可以作为一种绿色蔬菜,并日益引起重视。基于此,藜麦苗这一资源的深度开发和有效利用具有潜在的经济和社会价值。
多酚是植物中广泛存在的一类酚羟基结构的化合物,具有清除自由基、抗氧化、抑菌等多种功能活性,在食品、医药、保健品等领域具有广阔的应用前景。在藜麦的不同部位,多酚的含量也有所不同,且抗氧化能力也有差异。目前,现有技术对藜麦多酚类提取物的提取研究主要来源于籽粒,有关藜麦苗中多酚类物质的提取研究还很有限。根据现有的少量关于藜麦苗中多酚类物质提取的文献报道,提取工艺是使用溶剂浸提法,此法虽然操作简单,成本低,但是因细胞破碎不充分,有提取率低,耗时长等缺点,如果使用毒性较大的有机溶剂,则不利于安全生产和进一步应用。超临界或超高压辅助提取方法虽然可以提高提取率,但要求较高并增加了提取成本。这些问题的存在,限制了藜麦苗营养价值的全面开发利用。因此,亟需发展一种安全、高效、低成本的藜麦苗多酚提取工艺。
发明内容
针对现有技术问题,本发明提供一种藜麦苗中多酚类物质的提取工艺及应用。
为了解决上述技术问题,本发明采用的技术方案是:一种藜麦苗中多酚类物质的提取工艺及应用,包括以下步骤:
(1)将新鲜藜麦苗洗净,置于60℃烘箱内烘干至恒重,粉碎,过100目筛,得到藜麦苗粉末;
(2)称取藜麦苗粉末,加入一定量蒸馏水中,-25℃冷冻2h,60℃水浴融解10min,循环进行4-6次;
(3)在藜麦苗粉末水悬浮液中,加入无水乙醇形成乙醇水溶液作为提取溶剂,使乙醇水溶液体积百分比浓度为60%-80%,并且使藜麦苗粉末质量:乙醇水溶液体积为1:20,振荡混匀;
(4)超声提取,功率为300W,提取时间40-60min,提取温度50-70℃;提取后将粗提液以3500r/min离心15min,保留上清液;
(5)将上清液在45℃旋转蒸发浓缩成浸膏状,将浸膏状浓缩物冷冻干燥成粉末,制得藜麦苗多酚提取物。
进一步,对藜麦苗多酚含量进行测定,包括以下步骤:
(1)绘制标准曲线:精确称取没食子酸标准品0.0100g,用蒸馏水溶解并定容至100mL,得浓度为0.10mg/mL的标准液。准确吸取0、0.5、1.0、1.5、2.0、2.5mL置于25mL的棕色容量瓶中,加蒸馏水10mL,然后分别加入Folin-Ciocalteau试剂1mL,混匀,在0.5~8min内加入10mL 7%的Na2CO3溶液,充分混合后定容,得浓度为0、2、4、6、8、10μg/mL的没食子酸标准溶液,25℃下避光放置半小时,空白对照为不加标准液的6.0mL蒸馏水,在765nm波长下测定吸光值,以吸光值为纵坐标,没食子酸的浓度为横坐标绘制标准曲线(如图1所示)。
(2)藜麦苗多酚含量的测定:按照上述标准曲线实施步骤,对提取的藜麦苗多酚进行含量测定和计算。藜麦苗多酚含量以每克藜麦苗粉(干重)含有没食子酸当量的毫克数表示,单位为mg/g。
为获得藜麦苗多酚类物质最佳提取工艺,保持藜麦苗粉末质量:乙醇水溶液体积为1:20,超声提取功率300W,选取对藜麦苗多酚含量影响较显著的四个因素:乙醇浓度(A)、冻融次数(B)、提取时间(C)和提取温度(D)为自变量,以藜麦苗多酚含量为评价指标,采用正交试验法设计四因素三水平的分析试验,试验设计因素与水平如表1所示,试验结果如表2所示。表中数据结果表明,四个因素对藜麦苗多酚的含量影响由大到小依次为冻融次数>乙醇浓度>提取温度>提取时间,最优提取工艺条件为:乙醇浓度80%,冻融次数6次,超声提取时间50min,超声提取温度50℃。
表1试验设计因素与水平
表2正交试验结果
优选,所述步骤为:称取藜麦苗粉末,加入一定量蒸馏水中,-25℃冷冻2h,60℃水浴融解10min,循环冻融6次;在藜麦苗粉末水悬浮液中,加入无水乙醇形成乙醇水溶液作为提取溶剂,使乙醇水溶液体积百分比浓度为80%,并且使藜麦苗粉末质量:乙醇水溶液体积为1:20,振荡混匀;在50℃温度下,对藜麦苗粉末超声提取50min,超声功率为300W;将粗提液以3500r/min下离心15min,保存上清液;将上清液在45℃旋转蒸发浓缩成浸膏状,将浸膏状浓缩物冷冻干燥成粉末,制得藜麦苗多酚提取物。
本发明的有益效果是:采用本发明的方法可以对藜麦苗多酚类物质进行有效的提取,具有工艺简单、易于操作、提取率高、污染小、成本低等优点;本发明对藜麦苗粉末水悬浮液进行反复冻融处理,通过低温冷冻,使细胞中的水分瞬间凝结成冰晶,将细胞壁胀破;超声波具有空化作用、机械作用以及热效应;在冻融的基础上结合超声处理,可以最大限度地破坏藜麦苗细胞壁,使细胞内容物更彻底的溶出,因此提高提取率;选用的乙醇溶剂价格低廉,易于去除,无残留的毒副作用;本发明多酚的抗氧化效果,为将藜麦苗多酚应用在抗氧化制剂中提供了一种新的途径,可用于医药、食品及保健品领域。
附图说明
图1没食子酸标准曲线图。
图2藜麦苗多酚对DPPH自由基和羟基自由基的清除率图。
图3麦苗多酚对油脂氧化的抑制率图。
具体实施方式
下面结合具体实施例对本发明的实施方案作详细的说明。应该指出,以下详细说明都是例示性的,旨在对本申请提供进一步的说明。除非另有指明,本文使用的所有技术和科学术语具有与本申请所属技术领域的普通技术人员通常理解的相同含义。
本发明的一种藜麦苗多酚类物质的提取方法,将藜麦苗洗净,置于60℃烘箱中烘干至恒重并粉碎,过100目筛,得到藜麦苗粉末。称取藜麦苗粉末,加入一定量蒸馏水中,-25℃冷冻2h,60℃水浴融解10min,循环冻融4-6次;在藜麦苗粉末水悬浮液中,加入无水乙醇形成乙醇水溶液作为提取溶剂,使乙醇水溶液体积百分比浓度为60%-80%,并且使藜麦苗粉末质量:乙醇水溶液体积为1:20,振荡混匀;在50℃-70℃温度下,对藜麦苗粉末超声提取40-60min,超声功率为300W;将粗提液以3500r/min离心15min,保存上清液;将上清液在45℃旋转蒸发浓缩成浸膏状,将浸膏状浓缩物冷冻干燥成粉末,制得藜麦苗多酚类物质提取物。
其次,本发明还提供一种藜麦苗多酚类物质的抗氧化效果,此操作包括:取适量藜麦苗多酚提取物用水溶解,配制成一定质量浓度梯度的溶液,分别测定不同浓度下藜麦苗多酚提取物对DPPH自由基、羟基自由基的清除作用以及总还原能力。
进一步,本发明还提供一种藜麦苗多酚类物质提高油脂氧化稳定性的效果。此操作包括:以菜籽油质量为基准,按照不同浓度的添加量加入菜籽油中,均质处理,使藜麦苗多酚提取物均匀分散于菜籽油中,置于恒温振荡培养箱中进行加速氧化实验,定时测定菜籽油的丙二醛数值,计算氧化抑制率。
实施例1:称取1g藜麦苗粉末,加入到8mL蒸馏水中,振荡混匀,-25℃冷冻2h,然后迅速在60℃水浴融解10min,循环冻融4次;在藜麦苗粉末水悬浮液中,加入12mL无水乙醇形成乙醇水溶液作为提取溶剂,使乙醇水溶液体积百分比浓度为60%,并且使藜麦苗粉末质量:乙醇水溶液体积为1:20,振荡混匀;在50℃温度下,对藜麦苗粉末超声提取40min,超声功率为300W;将粗提液以3500r/min下离心15min,保存上清液;测得多酚含量为4.17mg/g。
实施例2:称取1g藜麦苗粉末,加入到8mL蒸馏水中,振荡混匀,-25℃冷冻2h,然后迅速在60℃水浴融解10min,循环冻融5次;在藜麦苗粉末水悬浮液中,加入12mL无水乙醇形成乙醇水溶液作为提取溶剂,使乙醇水溶液体积百分比浓度为60%,并且使藜麦苗粉末质量:乙醇水溶液体积为1:20,振荡混匀;在60℃温度下,对藜麦苗粉末超声提取50min,超声功率为300W;将粗提液以3500r/min下离心15min,保存上清液;测得多酚含量为4.43mg/g。
实施例3:称取1g藜麦苗粉末,加入到6mL蒸馏水中,振荡混匀,-25℃冷冻2h,然后迅速在60℃水浴融解10min,循环冻融6次;在藜麦苗粉末水悬浮液中,加入14mL无水乙醇形成乙醇水溶液作为提取溶剂,使乙醇水溶液体积百分比浓度为70%,并且使藜麦苗粉末质量:乙醇水溶液体积为1:20,振荡混匀;在60℃温度下,对藜麦苗粉末超声提取40min,超声功率为300W;将粗提液以3500r/min下离心15min,保存上清液;测得多酚含量为4.36mg/g。
实施例4:称取1g藜麦苗粉末,加入到6mL蒸馏水中,振荡混匀,-25℃冷冻2h,然后迅速在60℃水浴融解10min,循环冻融5次;在藜麦苗粉末水悬浮液中,加入14mL无水乙醇形成乙醇水溶液作为提取溶剂,使乙醇水溶液体积百分比浓度为70%,并且使藜麦苗粉末质量:乙醇水溶液体积为1:20,振荡混匀;在50℃温度下,对藜麦苗粉末超声提取60min,超声功率为300W;将粗提液以3500r/min下离心15min,保存上清液;测得多酚含量为4.22mg/g。
实施例5:称取1g藜麦苗粉末,加入到4mL蒸馏水中,振荡混匀,-25℃冷冻2h,然后迅速在60℃水浴融解10min,循环冻融5次;在藜麦苗粉末水悬浮液中,加入16mL无水乙醇形成乙醇水溶液作为提取溶剂,使乙醇水溶液体积百分比浓度为80%,并且使藜麦苗粉末质量:乙醇水溶液体积为1:20,振荡混匀;在70℃温度下,对藜麦苗粉末超声提取40min,超声功率为300W;将粗提液以3500r/min下离心15min,保存上清液;测得多酚含量为4.39mg/g。
实施例6:称取1g藜麦苗粉末,加入到4mL蒸馏水中,振荡混匀,-25℃冷冻2h,然后迅速在60℃水浴融解10min,循环冻融6次;在藜麦苗粉末水悬浮液中,加入16mL无水乙醇形成乙醇水溶液作为提取溶剂,使乙醇水溶液体积百分比浓度为80%,并且使藜麦苗粉末质量:乙醇水溶液体积为1:20,振荡混匀;在50℃温度下,对藜麦苗粉末超声提取50min,超声功率为300W;将粗提液以3500r/min下离心15min,保存上清液;测得多酚含量为4.72mg/g。
本发明进一步对藜麦苗多酚类物质进行抗氧化效果试验,试验涉及以下三个不同形式的效果体现:对DPPH自由基清除作用、对羟基自由基清除作用、总还原能力。
实施例1:藜麦苗多酚对DPPH自由基清除作用
取质量浓度为2,4,6,8,10μg/mL的藜麦苗多酚提取液2mL,分别加入0.2mmolDPPH·溶液2mL,摇匀,避光静置30min,以无水乙醇为空白在波长517nm处测定其吸光度A样品,用无水乙醇代替DPPH·溶液按上述方法测定吸光度A对照,用无水乙醇代替藜麦苗多酚提取液按上述方法测定的吸光度为A空白。按下式计算DPPH清除率。如图2所示,藜麦苗多酚类物质含量与DPPH自由基清除率成正比。
DPPH清除率/%=[1–(A样品–A对照)/A空白]×100%
实施例2:藜麦苗多酚对羟基自由基清除作用
在试管中依次加入2mL 6mmol/L FeSO4溶液,2mL质量浓度为2,4,6,8,10μg/mL的藜麦苗多酚提取液,2mL 6mmol/L H2O2溶液,摇匀,静置10min,再加入2mL 6mmol/L水杨酸溶液,摇匀,静置30min,以蒸馏水为空白于波长510nm处测定其吸光度A样品,用蒸馏水代替水杨酸按上述方法测定吸光度A对照,用蒸馏水代替多酚提取液按上述方法测定吸光度A空白。按下式计算羟基自由基的清除率。如图2所示,藜麦苗多酚类物质含量与羟基自由基清除率成正比。
羟基自由基清除率/%=[1–(A样品–A对照)/A空白]×100%
实施例3:藜麦苗多酚总还原能力
取质量浓度为2,4,6,8,10μg/mL的藜麦苗多酚提取液1mL,加入2.5mL pH 6.6的磷酸盐缓冲液和2.5mL 1%的铁氰化钾溶液,混匀。50℃保温20min,取出后加入2.5mL 10%的三氯乙酸,混匀。取出其中2.5mL,再加入2.5mL蒸馏水和0.5mL 0.1%三氯化铁溶液,混匀在波长700nm处测定吸光度。
本发明还对藜麦苗多酚类物质应用于延缓油脂氧化进行了试验,下面结合实施例进行说明:
实施例:藜麦苗多酚对油脂氧化抑制率的影响
丙二醛是油脂被氧化后的产物,它可以与TBA(硫代巴比妥酸)反应生成有色产物可在532nm下测定出其吸光值,吸光度值表示油脂氧化程度,其值越大,氧化程度越严重,当油脂中加入具有抗氧化作用的物质时,会减少氧化反应的发生从而降低丙二醛的含量。
以无水乙醇制备不同浓度藜麦多酚提取液,以猪油和菜籽油作为动物油和植物油样品。称取2g油样,分别按照0.25%、0.50%、0.75%、1.00%、1.25%和1.50%的添加量加入藜麦苗多酚提取液,在烘箱中45℃下恒温24h。取烘过的混合油样1mL加入2mL 20%的TCA(三氯乙酸)和2mL 0.6%的TBA在沸水浴下20min,取出冷却后加入5mL三氯甲烷,充分混匀,离心后取上清液比色,在532nm下测定其吸光度。按下式计算油脂的氧化抑制率。如图3所示,藜麦苗多酚类物质添加量与油脂氧化抑制率成正比。
抑制率/%=(A0-A1)/A0×100%
式中A0—以无水乙醇代替藜麦多酚提取物的吸光值;
A1—加入藜麦多酚提取物的吸光值。
以上所述仅为本申请的优选实施例而已,并不用于限制本申请。应当指出的是,对于本领域的技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形、改进及替代,这些都属于本发明的保护范围。
Claims (9)
1.一种藜麦苗多酚类物质的提取方法,其特征在于,包括以下步骤:
(1)将新鲜藜麦苗进行清洗、干燥、粉碎、过筛,得到藜麦苗粉末;
(2)称取藜麦苗粉末,加入到一定量蒸馏水中,振荡混匀,冷冻,然后迅速融解;在藜麦苗粉末水悬浮液中,加入无水乙醇,振荡混匀,形成乙醇水溶液作为提取溶剂,超声提取,然后离心,保留上清液;
(3)将上清液旋转蒸发浓缩至恒重,将浓缩物冷冻干燥成粉末,制得藜麦苗多酚类物质提取物。
2.根据权利要求1所述藜麦苗多酚类物质的提取方法,其特征在于,所述步骤(1)为将藜麦苗洗净,置于60℃烘箱内恒温烘干至恒重并粉碎,过100目筛。
3.根据权利要求1所述藜麦苗多酚类物质的提取方法,其特征在于,所述步骤(2)为-25℃冷冻2h,60℃水浴融解10min,循环进行4-6次。
4.根据权利要求1所述藜麦苗多酚类物质的提取方法,其特征在于,所述步骤(2)为加入无水乙醇使乙醇水溶液体积百分比浓度为60%-80%。
5.根据权利要求1所述藜麦苗多酚类物质的提取方法,其特征在于,所述步骤(2)为加入无水乙醇使藜麦苗粉末质量:乙醇水溶液体积为1:20。
6.根据权利要求1所述藜麦苗多酚类物质的提取方法,其特征在于,所述步骤(2)为超声提取的功率为300W,提取时间40-60min,提取温度50-70℃;提取后将粗提液以3500r/min离心15min。
7.根据权利要求1所述藜麦苗多酚的提取方法,其特征在于,所述步骤(3)为上清液旋转蒸发温度为45℃。
8.一种藜麦苗多酚类物质提取物,其特征在于:所述藜麦苗多酚类物质是根据权利要求1-7中任意一项所述的提取方法而得到的。
9.根据权利要求1-7所述的任一方法提取的藜麦苗多酚类物质在制备食品、保健品和医药产品中的应用。
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