CN110498785A - 一种黑枸杞花青素及其制备方法 - Google Patents
一种黑枸杞花青素及其制备方法 Download PDFInfo
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- CN110498785A CN110498785A CN201910835079.2A CN201910835079A CN110498785A CN 110498785 A CN110498785 A CN 110498785A CN 201910835079 A CN201910835079 A CN 201910835079A CN 110498785 A CN110498785 A CN 110498785A
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- Prior art keywords
- lycium ruthenicum
- anthocyanidin
- ruthenicum anthocyanidin
- preparation
- macroreticular resin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
- A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
- C07D311/60—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2
- C07D311/62—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with aryl radicals attached in position 2 with oxygen atoms directly attached in position 3, e.g. anthocyanidins
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
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Abstract
本发明公开了一种黑枸杞花青素及其制备方法,该制备方法包括以下步骤:黑枸杞花青素粉的制备;黑枸杞花青素提取;黑枸杞花青素的纯化及冷冻干燥。本发明制备得到的黑枸杞花青素有较好的清除DPPH自由基和羟自由基的能力;并对肥胖小鼠脂肪蓄积、脂质代谢、胰岛素耐受性的影响。
Description
技术领域
本发明属于天然产物开发技术领域,具体地说,涉及一种黑枸杞花青素及其制备方法。
背景技术
花青素(Anthocyanidin)又称花色素,是自然界广泛存在于植物中的一类水溶性的天然色素,属黄酮类化合物。花青素具有抗血脂、降血糖、抗氧化、免疫调节、延缓衰老、预防治疗心血管疾病、动脉硬化、肿瘤药物的应用价值。在清除自由基分子机制上类似于黄酮类化合物,可以清除体内和体外多种氧自由基,对体内各种抗氧化酶具有促进和激活作用;多酚结构可以络合金属离子作用,阻止此类具有氧化还原活性的金属离子发生催化作用以至加速自由基生成,最终加强其抗氧化作用。基于花青素的抗氧化活性,花青素发挥影响脂类代谢和降血糖的生理功能,还能抑制分裂素蛋白致活酶的活性,从而抑制肿瘤转移和生长。
黑枸杞(Lycium ruthenicum Murr)为茄科枸杞属植物,是一种野生珍稀植物资源,主要生长在海拔2700米-3000米的在青海德令哈等沙漠盆地地带,是我国西部地区特有的一种纯天然药食两用珍品,含有枸杞多糖、氨基酸、甜菜碱、胡萝卜素、维生素B、维生素C、烟酸等多种维生素,以及锰、铬、锌、铜、镁、钙、锗、钴等微量元素。更重要的是,黑枸杞富含花青素,被称为“花青素之王”,这是迄今为止发现花青素含量最高的天然野生果实,也是最有效的天然抗氧化剂,其功效是VC的20倍、VE的50倍,其具有预防心脑血管疾病,提高视力,降低血脂,抗癌、抗动脉粥样硬化、抗肿瘤、抗炎症、抗衰老、抗癌和抗辐射等功能,其维生素、矿物质等营养成分含量也很丰富,尤其含具有清除自由基、抗氧化功能的天然的花色甙素,药用,保健价值远远高于普通红枸杞,被誉为“软黄金”。
乙醇提取的黑枸杞花青素含有蛋白质、黄酮类化合物、多糖等杂质,可利用大孔树脂进行吸附纯化,提高黑枸杞花青素的含量和纯度。巩芳芳等利用体积分数70%酸性乙醇为提取液,采用超声波辅助法提取黑枸杞中的花青素,但此方法中的有机溶剂有一定的毒副作用,且产物提取率低。
发明内容
有鉴于此,本发明针对上述的问题,提供了一种黑枸杞花青素及其制备方法,本发明采取超声波辅助乙醇提取法对黑枸杞花青素进行提取,利用大孔树脂HZ-806进行吸附纯化,对黑枸杞花青素进行了清除DPPH自由基、羟自由基的测定和白色脂肪棕色化相关试验,试验结果均表明花青素具有良好的抗氧化性和促进白色脂肪棕色化。
为了解决上述技术问题,本发明公开了一种黑枸杞花青素的制备方法,包括以下步骤:
步骤1、黑枸杞花青素粉的制备;
步骤2、黑枸杞花青素提取;
步骤3、黑枸杞花青素的纯化及冷冻干燥。
可选地,所述步骤1中的黑枸杞花青素粉的制备具体为:挑选黑枸杞并去除杂物,于55-65℃下干燥10-15小时,然后经高速粉碎机粉碎,过100-150目筛,制备得到黑枸杞花青素粉,装袋备用。
可选地,所述步骤2中的黑枸杞花青素提取具体为:称取黑枸杞花青素干粉,按照质量体积比1:5-1:15的比例加入乙醇,于超声功率100-150w超声辅助提取20-30分钟,在5000-10000r/min下离心5-15min,取出上清液,并将滤渣重复提取,离心,合并提取液即为黑枸杞花青素提取液。
可选地,所述步骤3中的黑枸杞花青素的纯化及冷冻干燥具体为:将体积分数为95%的乙醇倒入装有大孔树脂的层析柱中,对大孔树脂进行浸泡,时间为20-28h,用蒸馏水对大孔树脂进行反复冲洗直至洗出液呈透明状,用体积分数为5%的HCl和体积分数为2%的NaOH分别浸泡大孔树脂3~5h,用蒸馏水洗至中性备用;用质量分数为60%的乙醇洗脱吸附于大孔树脂10-15h的花青素,将洗脱液装入瓶中进行浓缩,冷冻干燥得到粉末状的黑枸杞花青素。
本发明还公开了一种由上述的制备方法制备得到的黑枸杞花青素。
与现有技术相比,本发明可以获得包括以下技术效果:
1)黑枸杞花青素有较好的清除DPPH自由基的能力,清除率能达到78.23%;
2)黑枸杞花青素对羟自由基有一定的清除作用,清除率能达到23.35%;
3)黑枸杞花青素对肥胖小鼠脂肪蓄积、脂质代谢、胰岛素耐受性的影响。
当然,实施本发明的任一产品并不一定需要同时达到以上所述的所有技术效果。
附图说明
此处所说明的附图用来提供对本发明的进一步理解,构成本发明的一部分,本发明的示意性实施例及其说明用于解释本发明,并不构成对本发明的不当限定。在附图中:
图1是本发明黑枸杞花青素清除DPPH自由基的对比图;
图2是本发明黑枸杞花青素清除羟自由基的对比图;
图3是本发明普通日粮下黑枸杞花青素影响小鼠体重的对比图;
图4是本发明高脂饲料下黑枸杞花青素影响小鼠体重的对比图;
图5是本发明普通日粮下黑枸杞花青素影响葡萄糖耐受性的对比图;
图6是本发明高脂饲料下黑枸杞花青素影响葡萄糖耐受性的对比图;
图7是本发明普通日粮下黑枸杞花青素影响胰岛素耐受性的对比图;
图8是本发明高脂饲料下黑枸杞花青素影响胰岛素耐受性的对比图;
图9是本发明黑枸杞花青素对小鼠脂肪含量的影响;其中A代表棕色脂肪含量,B代表皮下脂肪含量,C代表内脏脂肪含量;
图10是本发明小鼠脂肪组织形态的变化;其中,a代表高脂饲料对照组,b代表高脂饲料花青素处理组,c代表普通日粮对照组,d代表普通日粮花青素处理组;右侧上图中的a代表高脂饲料对照组棕色脂肪,b代表高脂饲料花青素处理组棕色脂肪,c代表普通日粮对照组棕色脂肪,d代表普通日粮花青素处理组棕色脂肪;右侧下图中的a代表高脂饲料对照组皮下脂肪,b代表高脂饲料花青素处理组皮下脂肪,c代表普通日粮对照组皮下脂肪,d代表普通日粮花青素处理组皮下脂肪;
图11是本发明电镜下脂肪细胞中线粒体数量;
图12是本发明内脏脂肪相变化关细胞因子浓度的变化;
图13是本发明皮下脂肪相变化关细胞因子浓度的变化;
图14是本发明小鼠自适应产热相关蛋白的变化;A代表WB条带;B代表由条带做出的柱状图。
具体实施方式
以下将配合实施例来详细说明本发明的实施方式,藉此对本发明如何应用技术手段来解决技术问题并达成技术功效的实现过程能充分理解并据以实施。
实施例1
一种黑枸杞花青素的制备方法,包括以下步骤:
步骤1、黑枸杞花青素粉的制备:挑选黑枸杞并去除杂物,于60℃下干燥12小时,然后经高速粉碎机粉碎,过120目筛,制备得到黑枸杞花青素粉,装袋备用;
步骤2、黑枸杞花青素提取:称取黑枸杞花青素干粉,按照质量体积比1:10的比例加入乙醇,于超声功率120w超声辅助提取25分钟,在8000r/min下离心10min,取出上清液,并将滤渣重复提取,离心,合并提取液即为黑枸杞花青素提取液。
步骤3、黑枸杞花青素的纯化及冷冻干燥:将体积分数为95%的乙醇倒入装有大孔树脂的层析柱中,对大孔树脂进行浸泡,时间为24h,用蒸馏水对大孔树脂进行反复冲洗直至洗出液呈透明状,用体积分数为5%的HCl和体积分数为2%的NaOH分别浸泡大孔树脂4h,用蒸馏水洗至中性备用;用体积分数为60%的乙醇洗脱吸附于大孔树脂12h的花青素,将洗脱液装入瓶中进行浓缩,冷冻干燥得到粉末状的黑枸杞花青素。
实施例2
一种黑枸杞花青素的制备方法,包括以下步骤:
步骤1、黑枸杞花青素粉的制备:挑选黑枸杞并去除杂物,于55℃下干燥15小时,然后经高速粉碎机粉碎,过100目筛,制备得到黑枸杞花青素粉,装袋备用;
步骤2、黑枸杞花青素提取:称取黑枸杞花青素干粉,按照质量体积比1:15的比例加入乙醇,于超声功率100w超声辅助提取30分钟,在5000r/min下离心15min,取出上清液,并将滤渣重复提取,离心,合并提取液即为黑枸杞花青素提取液。
步骤3、黑枸杞花青素的纯化及冷冻干燥:将质量分数为95%的乙醇倒入装有大孔树脂的层析柱中,对大孔树脂进行浸泡,时间为20h,用蒸馏水对大孔树脂进行反复冲洗直至洗出液呈透明状,用质量分数为5%的HCl和质量分数为2%的NaOH分别浸泡大孔树脂5h,用蒸馏水洗至中性备用;用质量分数为60%的乙醇洗脱吸附于大孔树脂10h的花青素,将洗脱液装入瓶中进行浓缩,冷冻干燥得到粉末状的黑枸杞花青素。
实施例3
一种黑枸杞花青素的制备方法,包括以下步骤:
步骤1、黑枸杞花青素粉的制备:挑选黑枸杞并去除杂物,于65℃下干燥10小时,然后经高速粉碎机粉碎,过150目筛,制备得到黑枸杞花青素粉,装袋备用;
步骤2、黑枸杞花青素提取:称取黑枸杞花青素干粉,按照质量体积比1:5的比例加入乙醇,于超声功率150w超声辅助提取20分钟,在10000r/min下离心5min,取出上清液,并将滤渣重复提取,离心,合并提取液即为黑枸杞花青素提取液。
步骤3、黑枸杞花青素的纯化及冷冻干燥:将质量分数为95%的乙醇倒入装有大孔树脂的层析柱中,对大孔树脂进行浸泡,时间为28h,用蒸馏水对大孔树脂进行反复冲洗直至洗出液呈透明状,用质量分数为5%的HCl和质量分数为2%的NaOH分别浸泡大孔树脂3h,用蒸馏水洗至中性备用;用质量分数为60%的乙醇洗脱吸附于大孔树脂15h的花青素,将洗脱液装入瓶中进行浓缩,冷冻干燥得到粉末状的黑枸杞花青素。
实施例4
一种黑枸杞花青素的制备方法,包括以下步骤:
步骤1、黑枸杞花青素粉的制备:挑选黑枸杞并去除杂物,于58℃下干燥14小时,然后经高速粉碎机粉碎,过140目筛,制备得到黑枸杞花青素粉,装袋备用;
步骤2、黑枸杞花青素提取:称取黑枸杞花青素干粉,按照质量体积比1:12的比例加入乙醇,于超声功率110w超声辅助提取22分钟,在6000r/min下离心12min,取出上清液,并将滤渣重复提取,离心,合并提取液即为黑枸杞花青素提取液。
步骤3、黑枸杞花青素的纯化及冷冻干燥:将质量分数为95%的乙醇倒入装有大孔树脂的层析柱中,对大孔树脂进行浸泡,时间为22h,用蒸馏水对大孔树脂进行反复冲洗直至洗出液呈透明状,用质量分数为5%的HCl和质量分数为2%的NaOH分别浸泡大孔树脂4h,用蒸馏水洗至中性备用;用质量分数为60%的乙醇洗脱吸附于大孔树脂14h的花青素,将洗脱液装入瓶中进行浓缩,冷冻干燥得到粉末状的黑枸杞花青素。
实施例5黑枸杞花青素清除DPPH自由基
将实施例1制备得到的黑果枸杞花青素提取物粉末进行溶解,并配制成5种浓度,每种浓度各取1m L分别放入试管中,并分别加入80μmol/L的DPPH·溶液5m L,混合均匀,避光保存20min,在517nm处,对混合液进行吸光度的测定,记为Ai;取1.0m L的蒸馏水放入试管中,加入80μmol/L的DPPH·溶液5m L,摇晃均匀后,避光保存20min,在517nm处,对其进行吸光度测定,记为AC;对照组将VC粉末进行溶解,代替样品对其进行测定,步骤同上,按下列公式计算清除率。
清除率(%)=((Ac—Ai)/Ac)×100
式中:Ac为蒸馏水加DPPH·溶液的吸光度;
Ai为不同浓度样品加DPPH·溶液的吸光度。
实验结果见图1,黑枸杞花青素组对DPPH自由基的清除率显著高于对照组,黑枸杞花青素增强小鼠清除DPPH自由基的能力。
实施例6黑枸杞花青素清除羟自由基:
将实施例1制备得到的黑果枸杞花青素提取物粉末进行溶解,并配制成5种浓度,每种浓度各取1m L分别放入试管中,在各试管中分别加入配制好的Fe2SO4,其浓度为8000μmol/L和以无水乙醇配制的8000μmol/L的水杨酸各1m L,反复摇晃试管,直至溶液混合均匀,然后将8.8mmol/L的H2O2加入试管中,每个试管1m L,在恒温为38℃的水浴中放置40min,使其充分反应。以超纯水为空白对照品和参比液,对照品组以VC溶液测定,步骤同上,在520nm处,对混合液进行吸光度的测定,按下列公式计算清除率:
羟基自由基的清除率(%)=((A0—A1)/A0)×100
式中:A0、A1分别为空白与样品的平均吸光度值。
实验结果见图2,黑枸杞花青素组对羟自由基的清除率显著高于对照组,黑枸杞花青素增强小鼠清除羟自由基的能力。
实施例7黑枸杞花青素对肥胖小鼠脂肪蓄积、脂质代谢、胰岛素耐受性的影响
1、黑枸杞花青素对小鼠及肥胖小鼠体质量的影响
以8周龄C57BL/6小鼠为实验动物,将小鼠分为普通日粮组、普通日粮花青素添加组(高剂量组、中剂量组、低剂量组)、高脂组和高脂添加花青素(高剂量组、中剂量组、低剂量组)六组。从第8周开始饲喂不同饲粮,同时每周测量小鼠体重,观测添加过程中花青素对小鼠体重和高脂诱导的体重增长的影响,连续检测8周。花青素的给药方式为灌胃给药。
实验结果见图3和图4,饲喂普通日粮情况下,第七周花青素添加组小鼠体重与对照组小鼠体重有差异;饲喂高脂饲料情况下,第五周花青素添加组小鼠体重与对照组小鼠体重有差异,说明黑枸杞花青素可降低小鼠体重,尤其对高脂诱导的体重增长有明显减缓作用。
2、黑枸杞花青素对普通小鼠和肥胖小鼠空腹血糖、葡萄糖和胰岛素耐受性影响
测定空腹血糖,将小鼠先禁食12~16h,然后测定空腹血糖水平,随后做GTT测试,即给小鼠腹腔注射质量浓度为2g/kg的葡萄糖溶液,然后在注射后0、15、30、60、90、120min等6个时间点依次测定血糖值,比较各处理组间葡萄糖耐受的差别,研究花青素对小鼠糖耐受性变化的影响。恢复一周后,再做ITT测试,即禁食4h后测定血糖值,然后给小鼠腹腔注射1.5U/kg胰岛素,然后测定0、30、60、90、120min等5个时间点的血糖值。
实验结果见图5-图8,由图可知,无论普通日粮还是高脂饲料,黑枸杞花青素可明显改善小鼠的葡萄糖和胰岛素的耐受。
3、黑枸杞花青素对小鼠脂肪分布的的影响
(1)监测活化的棕色脂肪含量(采用DUEX正电子螺旋CT扫描(PET-CT)技术检测F18标记的葡萄糖示踪技术,扫描并计算小鼠棕色脂肪和白色色脂肪的体表和体内分布含量差异)。
(2)脱颈椎处死小鼠,迅速剥离小鼠肩胛骨间隙、肾周脂肪、附睾脂肪和皮下脂肪拍照并称重记录。明确小鼠脂肪的分布情况;取上述组织进行HE染色和油红O染色,观察组织中脂肪细胞的形态变化;电镜下观察脂肪细胞中线粒体数量。
实验结果见图9-图11,黑枸杞花青素显著对小鼠棕色脂肪和皮下脂肪重量的影响不显著,但降低小鼠皮下脂肪的重量;黑枸杞花青素处理组组织中脂肪细胞变小;电镜下观察脂肪细胞中线粒体数量,黑枸杞花青素处理组线粒体数量增加。
4、黑枸杞花青素对小鼠脂肪中与脂肪代谢UCP1等相关基因mRNA和蛋白表达的影响
脂肪沉积相关因子:检测影响脂肪细胞分化因子以及脂代谢相关蛋白的表达:如UCP1、LPL、SCD1、FAS、ACS、HSL、PPARγ、C/EBPs、TNF-α等的表达。具体如下:
(1)Real-time PCR检测相关基因表达
小鼠附睾脂肪组织中总RNA的提取按照试剂盒进行,以琼脂糖凝胶检测RNA品质;按试剂盒合成cDNA第一链,-40℃保存。
总RNA提取:组织中总RNA的提取:取0.5g冻存的组织块在装有液氮的研钵中快速研磨成粉末状,待液氮挥发完全后,用干净小勺把粉末放入盛有1mL RNAiso Plus Reagent的离心管中,剧烈振荡5min,然后室温静置15min;从细胞中提取总RNA。严格按照Real-timePCR操作说明提取RNA,以合格RNA为模板反转录合成cDNA第一链,用于PCR扩增模板。反转录按照说明进行操作,合成的cDNA。
Real-time PCR反应使用TaKaRa公司SYBR Premix ExTaqII试剂盒,利用Biorad公司iQ5型实时定量PCR系统进行检测。
(2)Westernblotting检测相关蛋白表达
1)组织中总蛋白的提取:取1g左右冷冻保存的组织样品研磨成粉末,然后盛有蛋白裂解液的离心管中。振荡混匀后离心10min。吸取上清液转移至新的离心管中,-76℃保存或继续操作。
2)细胞中总蛋白的提取:在细胞间内,将培养皿中的培养液吸出,PBS溶液冲洗细3次,用胰酶将贴壁的细胞消化下来,离心、裂解、离心后-76℃保存或继续操作。
蛋白浓度的检测:
3)制作标准曲线
4)检测样品蛋白含量。(按照Westernblotting实验方法进行)。
实验结果见图12-图13。
5、对小鼠PI3K、PPARγ信号通路关键因子mRNA和蛋白表达的影响,方法同上(4)。实验结果见图14。
上述说明示出并描述了发明的若干优选实施例,但如前所述,应当理解发明并非局限于本文所披露的形式,不应看作是对其他实施例的排除,而可用于各种其他组合、修改和环境,并能够在本文所述发明构想范围内,通过上述教导或相关领域的技术或知识进行改动。而本领域人员所进行的改动和变化不脱离发明的精神和范围,则都应在发明所附权利要求的保护范围内。
Claims (5)
1.一种黑枸杞花青素的制备方法,其特征在于,包括以下步骤:
步骤1、黑枸杞花青素粉的制备;
步骤2、黑枸杞花青素提取;
步骤3、黑枸杞花青素的纯化及冷冻干燥。
2.根据权利要求1所述的制备方法,其特征在于,所述步骤1中的黑枸杞花青素粉的制备具体为:挑选黑枸杞并去除杂物,于55-65℃下干燥10-15小时,然后经高速粉碎机粉碎,过100-150目筛,制备得到黑枸杞花青素粉,装袋备用。
3.根据权利要求1所述的制备方法,其特征在于,所述步骤2中的黑枸杞花青素提取具体为:称取黑枸杞花青素干粉,按照质量体积比1:5-1:15的比例加入乙醇,于超声功率100-150w超声辅助提取20-30分钟,在5000-10000r/min下离心5-15min,取出上清液,并将滤渣重复提取,离心,合并提取液即为黑枸杞花青素提取液。
4.根据权利要求1所述的制备方法,其特征在于,所述步骤3中的黑枸杞花青素的纯化及冷冻干燥具体为:将体积分数为95%的乙醇倒入装有大孔树脂的层析柱中,对大孔树脂进行浸泡,时间为20-28h,用蒸馏水对大孔树脂进行反复冲洗直至洗出液呈透明状,用体积分数为5%的HCl和体积分数为2%的NaOH分别浸泡大孔树脂3~5h,用蒸馏水洗至中性备用;用质量分数为60%的乙醇洗脱吸附于大孔树脂10-15h的花青素,将洗脱液装入瓶中进行浓缩,冷冻干燥得到粉末状的黑枸杞花青素。
5.一种由权利要求1-4中任一权利要求所述的制备方法制备得到的黑枸杞花青素。
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Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111513147A (zh) * | 2020-04-24 | 2020-08-11 | 上海应用技术大学 | 一种天然猪油抗氧化剂及制备方法 |
CN111647283A (zh) * | 2020-06-04 | 2020-09-11 | 宁夏北方生物科技有限公司 | 一种枸杞红色素的提取方法 |
CN111777585A (zh) * | 2020-07-24 | 2020-10-16 | 哈工大机器人南昌智能制造研究院 | 一种高纯度黑枸杞花青素粉末的制备方法 |
CN111875573A (zh) * | 2020-07-24 | 2020-11-03 | 哈工大机器人南昌智能制造研究院 | 一种连续提取黑枸杞中花青素的方法 |
CN117338856A (zh) * | 2023-09-28 | 2024-01-05 | 新疆农垦科学院 | 一种黑枸杞花色苷组合物的制备方法和产品及其应用 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101912480A (zh) * | 2010-07-23 | 2010-12-15 | 中国科学院西北高原生物研究所 | 一种黑果枸杞原花青素提取物的制备方法 |
CN103483304A (zh) * | 2013-05-20 | 2014-01-01 | 宁夏润丰枸杞生物制品有限公司 | 从黑枸杞中提取花青素的工艺方法 |
CN104940038A (zh) * | 2015-05-28 | 2015-09-30 | 秦春哲 | 黑枸杞花青素在制备化妆品中的应用及含有黑枸杞花青素的化妆品 |
CN106905390A (zh) * | 2017-03-03 | 2017-06-30 | 甘肃中医药大学 | 一种黑果枸杞花青素的制备工艺及其检测方法与用途 |
CN109574979A (zh) * | 2018-12-28 | 2019-04-05 | 中国科学院西北高原生物研究所湖州高原生物资源产业化创新中心 | 一种黑果枸杞蓝色花青素及其制备方法 |
-
2019
- 2019-09-05 CN CN201910835079.2A patent/CN110498785A/zh active Pending
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101912480A (zh) * | 2010-07-23 | 2010-12-15 | 中国科学院西北高原生物研究所 | 一种黑果枸杞原花青素提取物的制备方法 |
CN103483304A (zh) * | 2013-05-20 | 2014-01-01 | 宁夏润丰枸杞生物制品有限公司 | 从黑枸杞中提取花青素的工艺方法 |
CN104940038A (zh) * | 2015-05-28 | 2015-09-30 | 秦春哲 | 黑枸杞花青素在制备化妆品中的应用及含有黑枸杞花青素的化妆品 |
CN106905390A (zh) * | 2017-03-03 | 2017-06-30 | 甘肃中医药大学 | 一种黑果枸杞花青素的制备工艺及其检测方法与用途 |
CN109574979A (zh) * | 2018-12-28 | 2019-04-05 | 中国科学院西北高原生物研究所湖州高原生物资源产业化创新中心 | 一种黑果枸杞蓝色花青素及其制备方法 |
Non-Patent Citations (4)
Title |
---|
任小娜等: "黑果枸杞中原花青素稳定性和抗氧化性研究", 《湖北农业科学》 * |
杭园园等: "黑果枸杞花青素制备及其抗氧化活性研究", 《食品科技》 * |
杭园园等: "黑果枸杞花青素的制备与纯化研究", 《食品安全质量检测学报》 * |
赵文娟等: "大孔吸附树脂纯化黑果枸杞中的原花青素", 《食品工业科技》 * |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111513147A (zh) * | 2020-04-24 | 2020-08-11 | 上海应用技术大学 | 一种天然猪油抗氧化剂及制备方法 |
CN111647283A (zh) * | 2020-06-04 | 2020-09-11 | 宁夏北方生物科技有限公司 | 一种枸杞红色素的提取方法 |
CN111777585A (zh) * | 2020-07-24 | 2020-10-16 | 哈工大机器人南昌智能制造研究院 | 一种高纯度黑枸杞花青素粉末的制备方法 |
CN111875573A (zh) * | 2020-07-24 | 2020-11-03 | 哈工大机器人南昌智能制造研究院 | 一种连续提取黑枸杞中花青素的方法 |
CN117338856A (zh) * | 2023-09-28 | 2024-01-05 | 新疆农垦科学院 | 一种黑枸杞花色苷组合物的制备方法和产品及其应用 |
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