CN113332453B - 水溶性蜂胶纳米颗粒的制备方法 - Google Patents

水溶性蜂胶纳米颗粒的制备方法 Download PDF

Info

Publication number
CN113332453B
CN113332453B CN202110724960.2A CN202110724960A CN113332453B CN 113332453 B CN113332453 B CN 113332453B CN 202110724960 A CN202110724960 A CN 202110724960A CN 113332453 B CN113332453 B CN 113332453B
Authority
CN
China
Prior art keywords
propolis
water
solution
nano
soluble
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN202110724960.2A
Other languages
English (en)
Other versions
CN113332453A (zh
Inventor
郝玉徽
贺莹娟
冉永红
高继宁
杨娟
柳随义
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Third Military Medical University TMMU
Original Assignee
Third Military Medical University TMMU
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Third Military Medical University TMMU filed Critical Third Military Medical University TMMU
Priority to CN202110724960.2A priority Critical patent/CN113332453B/zh
Publication of CN113332453A publication Critical patent/CN113332453A/zh
Application granted granted Critical
Publication of CN113332453B publication Critical patent/CN113332453B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/58—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. poly[meth]acrylate, polyacrylamide, polystyrene, polyvinylpyrrolidone, polyvinylalcohol or polystyrene sulfonic acid resin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/56—Materials from animals other than mammals
    • A61K35/63—Arthropods
    • A61K35/64—Insects, e.g. bees, wasps or fleas
    • A61K35/644—Beeswax; Propolis; Royal jelly; Honey
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
    • A61K47/59—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes
    • A61K47/60—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyureas or polyurethanes the organic macromolecular compound being a polyoxyalkylene oligomer, polymer or dendrimer, e.g. PEG, PPG, PEO or polyglycerol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
    • A61K47/6927—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores
    • A61K47/6929—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
    • A61K47/6931—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer
    • A61K47/6933—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer the polymer being obtained by reactions only involving carbon to carbon, e.g. poly(meth)acrylate, polystyrene, polyvinylpyrrolidone or polyvinylalcohol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
    • A61K47/6921—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere
    • A61K47/6927—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores
    • A61K47/6929—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle
    • A61K47/6931—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer
    • A61K47/6935—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit the form being a particulate, a powder, an adsorbate, a bead or a sphere the form being a solid microparticle having no hollow or gas-filled cores the form being a nanoparticle, e.g. an immuno-nanoparticle the material constituting the nanoparticle being a polymer the polymer being obtained otherwise than by reactions involving carbon to carbon unsaturated bonds, e.g. polyesters, polyamides or polyglycerol
    • 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
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Veterinary Medicine (AREA)
  • Epidemiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Nanotechnology (AREA)
  • Insects & Arthropods (AREA)
  • Immunology (AREA)
  • Biochemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Animal Husbandry (AREA)
  • Toxicology (AREA)
  • Zoology (AREA)
  • Jellies, Jams, And Syrups (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

本发明涉及一种水溶性蜂胶纳米颗粒的制备方法，该方法是，取20‑150mg蜂胶溶解与质量百分含量为0.5%—2%碱性溶液，得到蜂胶碱性溶液，搅拌下快速加入到含有高分子交联剂的酸性溶液中，磁力搅拌反应2小时，在10000转/分的条件下离心20min分离出固体产物，水洗3次，得到水溶性蜂胶纳米颗粒。该方法不使用有机溶剂，利用蜂胶中的成分与交联剂中的羟基等形成结构稳定的氢键，制备蜂胶纳米颗粒水溶性好、稳定性好、分散性好，该方法制备的蜂胶纳米颗粒保持了蜂胶的生物活性，具有优异的抗氧化性能。

Description

水溶性蜂胶纳米颗粒的制备方法
技术领域
本发明涉及一种纳米药物的制备方法，特别涉及一种水溶性蜂胶纳米颗粒的制备方法。
背景技术
蜂胶是一种传统的天然药物，是工蜂从植物的树芽以及树皮上采集的树脂，并混入蜜蜂的上腭腺体的分泌物，经蜜蜂加工转化而成的具有芳香性不透明的胶状物质，用于维持整个群体健康的有效物质，也是蜜蜂防止外来入侵者的“化学武器”。
蜂胶成分相当复杂，其主要成分是黄酮类化合物、萜类化合物、酚类、醇类、有机酸、酯类以及多种氨基酸、酶、微生物等。
蜂胶具有广泛的生物学活性和药理活性。国内外大量研究证明，蜂胶具有抗氧化活性、广谱的抗菌消炎作用、免疫调节活性、口腔保健作用、创口愈合作用。但是由于蜂胶的主要功效成分为黄酮类和萜烯类化合物很难溶于水而常溶于有机溶剂，现有的蜂胶产品主要是蜂胶软胶囊、复方蜂胶片、纳米乳以及蜂胶纳米颗粒。这些产品都存在一些不足，片制剂加入了大量其他助剂进行加工，剂型不稳定、吸收效果不好。
通过纳米技术将蜂胶制成纳米尺寸的纳米颗粒不仅可以提高蜂胶的生物利用率，还可以提高其水溶性。现有蜂胶纳米粒（例如专利申请号201911044981）制备过程需要加入有机溶剂（丙酮、甲醇等），会破坏蜂胶活性成分，且稳定性不好，制备受超声能量的影响比较大。专利200510049745中提到的纳米蜂胶制品需要加入卵磷脂等脂质体对蜂胶进行包裹负载，载药率有限，所得蜂胶纳米颗粒水溶性不好，限制了蜂胶的使用，且复杂的制备过程会降低蜂胶的生物活性。
发明内容
本发明的目的在于克服现有技术中其它蜂胶产品存在的缺陷，提供一种水溶性蜂胶纳米颗粒的制备方法，该方法不使用有机溶剂，制备出的蜂胶纳米颗粒水溶性好、分散性好，具有优异的抗氧化性。
本发明的技术方案是：
水溶性蜂胶纳米颗粒的制备方法取20-150mg蜂胶溶解与质量百分含量为0.5%—2%碱性溶液，得到蜂胶碱性溶液，搅拌下快速加入到含有高分子交联剂的酸性溶液中，磁力搅拌反应2小时，在10000转/分的条件下离心20min分离出固体产物，水洗3次，得到水溶性蜂胶纳米颗粒。
所述高分子交联剂的质量百分比浓度为0.01%—0.1%。
蜂胶与高分子交联剂的质量比为20:1—100:1。
所述碱性溶液是质量百分含量为0.1%—1%的氢氧化钠溶液。
所述酸性溶液的质量百分含量为0.5%—2%；其酸性溶液可以是醋酸溶液，柠檬酸溶液中的任一种或者是两种溶液的混合。
高分子交联剂选自聚乙烯醇、聚乙烯吡咯烷酮、泊洛沙姆188中的任一种。
聚乙烯醇的平均分子量范围为13000—98000，聚乙烯吡咯烷酮平均分子量范围为8000—29000。
磁力搅拌速率采用400转/分的速率快速搅拌5分钟后，以50转/分的速率低速搅拌反应2小时。
所述水溶性蜂胶纳米颗粒的平均粒径为100—200nm，分散系数PDI小于0.1，表面电位为-2mV— -10mV（参见图5）。
本发明利用高分子交联剂中的羟基与蜂胶中的羟基之间的范德华力，能自组装形成稳定的、水溶性的蜂胶纳米颗粒，有效解决蜂胶水溶性差以及由其带来的难以分散、难以给药等一系列问题，避免制备过程中加入的有机溶剂对环境的污染，提高蜂胶的应用价值，利于工业化。所述水溶性的蜂胶纳米颗粒平均粒径为100—200nm，分散系数PDI小于0.1，表面电位为-2mV— -10mV。傅里叶红外光谱表征可以看出蜂胶与高分子交联剂的之间产生了物理交联，出现了了氢键特征峰(参见图4)。
本发明所述制备方法，利用蜂胶溶于碱性溶液而不溶于酸性溶液的性质，析出来的蜂胶与酸性溶液中的高分子交联剂自组装形成蜂胶纳米颗粒，最后通过高速离心机将水溶性蜂胶纳米颗粒分离出来。本发明采用类似纳米沉降方法快速制备出水溶性蜂胶纳米颗粒，有效解决了蜂胶水溶性差、不稳定及其带来的难以给药、难以分散等问题。方法简单、不使用有机污染物，保留了蜂胶的生物学效应。
该方法制备出来的蜂胶纳米颗粒保留了蜂胶的良好的抗氧化性生物活性。
附图说明
图1为实施例1制备的水溶性蜂胶纳米颗粒的粒径分布曲线；
图2为实施例2制备的水溶性蜂胶纳米颗粒的粒径分布曲线；
图3为实施例3制备的水溶性蜂胶纳米颗粒的粒径分布曲线；
图4为实施例1制备的水溶性蜂胶纳米颗粒的红外谱图；
图5为水溶性蜂胶纳米颗粒的透射电镜图。
具体实施方式
本实施例除蜂胶外其他试剂均采用市售的分析纯产品。
实施例1
将50mg蜂胶（闻达生物，药品级别）溶解于10 mL质量分数0.5%的氢氧化钠（国药集团化学试剂有限公司，分析纯）溶液中，溶解完全后，一边搅拌一边快速将上述溶液加入到20 mL质量分数为1%的醋酸（国药集团化学试剂有限公司，分析纯）溶液中（醋酸溶液中含有10mg的聚乙烯醇13000-23000（sigma试剂，分析纯）)，50转/分磁力搅拌反应2小时，10000转/分的条件下离心20min分离出固体产物，水洗3次后得到水溶性蜂胶纳米颗粒。平均粒径约为143nm，分散系数PDI=0.059,表面电位为-7mV，蜂胶纳米颗粒的粒径分布曲线参见图1。
实施例2
将100mg蜂胶溶解于5 mL质量分数0.1%的氢氧化钠溶液中，溶解完全后，一边搅拌一边快速将上述溶液加入到30 mL质量分数为1%的醋酸溶液中（醋酸溶液中含有6mg的泊洛沙姆188（sigma试剂，分析纯）)，50转/分磁力搅拌反应2小时，10000转/分的条件下离心20min分离出固体产物，水洗3次后得到水溶性蜂胶纳米颗粒。平均粒径约为126nm，分散系数PDI=0.076，表面电位为-3mV。蜂胶纳米颗粒的粒径分布曲线参见图2
实施例3
将80mg蜂胶溶解于10 mL质量分数0.3%的氢氧化钠溶液中，溶解完全后，一边搅拌一边快速将上述溶液加入到50 mL质量分数为1%的柠檬酸钠溶液中（柠檬酸钠溶液中含有20mg的聚吡咯烷酮29000)，50转/分磁力搅拌反应2小时，10000转/分的条件下离心20min分离出固体产物，水洗3次后得到水溶性蜂胶纳米颗粒。平均粒径约为117nm，分散系数PDI=0.047,表面电位为-9mV。蜂胶纳米颗粒的粒径分布曲线参见图3
实施例4 DPPH法评价水溶性蜂胶纳米颗粒的抗氧化性能
取实施例1所得水溶性蜂胶纳米颗粒，加入超纯水配制成0.05mg/mL的水溶性蜂胶纳米颗粒溶液，取500uL上述水溶性蜂胶纳米颗粒溶液加入到5mL 0.1mM的DPPH乙醇溶液中，混匀后25℃避光放置1h，采用UV-3600在517nm处测吸光度。对照组加入500uL超纯水。水溶性蜂胶纳米颗粒对DPPH的清除率为（3100.89±128.02）umol/g。
实施例5 ABTS法评价水溶性蜂胶纳米颗粒的抗氧化性能
取实施例2所得水溶性蜂胶纳米颗粒，加入超纯水配制成7.5ug/mL的水溶性蜂胶纳米颗粒溶液，将2mM的ABTS溶液与2.45mM过硫酸钾(K2S2O8)混合溶解在PBS溶液(25 mM,pH 7.4)中制备成ABTS·+溶液，混匀后25℃避光放置4h，随后将上述溶液用PBS稀释，直到溶液在734nm处吸光度值为0.7±0.05。将500uL 7.5ug/mL水溶性蜂胶纳米颗粒溶液加入到4mL稀释后的ABTS·+溶液中，30min后用UV3600测定吸光度值ABTS自由基清除率，对照组加入500uL超纯水。水溶性蜂胶纳米颗粒对ABTS自由基清除率为（273.86±11.22）mmol/g。
实施例6 FRAP法评价水溶性蜂胶纳米颗粒的抗氧化性能
取实施例3所得水溶性蜂胶纳米颗粒，加入超纯水配制成0.05mg/mL的水溶性蜂胶纳米颗粒溶液，将3mol/L醋酸缓冲液（pH=3.6），0.1mol/L TPTZ以及0.2mol/L FeCl3.6H2O按照10:1:1的体积比混合制备出FRAP工作液（现配现用），37℃反应60min。将500uL 0.05mg/mL水溶性蜂胶纳米颗粒溶液加入到5mLFRAP工作液中，37℃反应15min，在593nm处测吸光度值。水溶性蜂胶纳米颗粒还原Fe3+的能力为（27.19±1.07）mmol/g。
结论： 采用本专利中的方法能快速制备出水溶性蜂胶纳米颗粒，利用蜂胶与高分子交联剂之间的氢键作用使得蜂胶纳米颗粒具有较好的水溶性、稳定性。有效解决了蜂胶水溶性差、不稳定及其带来的难以给药、难以分散等问题。制备方法简单、不使用有机污染物，保留了蜂胶的生物学效应，具有较强的抗氧化性能。

Claims (8)

1.一种水溶性蜂胶纳米颗粒的制备方法，其特征在于：取20-150mg蜂胶溶解于质量百分含量为0.5%—2%的碱性溶液，得到蜂胶碱性溶液，搅拌下快速加入到含有高分子交联剂的酸性溶液中，磁力搅拌反应2小时，随后在10000转/分的条件下离心20min分离出固体产物，水洗3次，得到水溶性蜂胶纳米颗粒；
高分子交联剂选自聚乙烯醇、聚乙烯吡咯烷酮、泊洛沙姆188中的任一种，所述方法不使用有机溶剂。
2.根据权利要求1所述的方法，其特征在于，所述高分子交联剂的质量百分比浓度为0.01%—0.1%。
3.根据权利要求1所述的方法，其特征在于，所述碱性溶液是质量百分含量为0.5%—1%的氢氧化钠溶液。
4.根据权利要求1所述的方法，其特征在于，所述酸性溶液的质量百分含量为0.5%—2%；其酸性溶液选自醋酸溶液，柠檬酸溶液中的任一种。
5.根据权利要求1所述的方法，其特征在于，聚乙烯醇的平均分子量范围为13000—98000，聚乙烯吡咯烷酮平均分子量范围为8000—29000。
6.根据权利要求1所述的方法，其特征在于，蜂胶碱性溶液与酸性溶液的体积比为1:2—1:7。
7.根据权利要求1所述的方法，其特征在于，磁力搅拌速率采用50转/分的速率低速搅拌反应2小时。
8.根据权利要求1所述的方法，其特征在于，所述水溶性蜂胶纳米颗粒的平均粒径为100—200nm，分散系数PDI小于0.1，表面电位为-2mV—-10mV。
CN202110724960.2A 2021-06-29 2021-06-29 水溶性蜂胶纳米颗粒的制备方法 Active CN113332453B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202110724960.2A CN113332453B (zh) 2021-06-29 2021-06-29 水溶性蜂胶纳米颗粒的制备方法

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202110724960.2A CN113332453B (zh) 2021-06-29 2021-06-29 水溶性蜂胶纳米颗粒的制备方法

Publications (2)

Publication Number Publication Date
CN113332453A CN113332453A (zh) 2021-09-03
CN113332453B true CN113332453B (zh) 2022-09-20

Family

ID=77481339

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202110724960.2A Active CN113332453B (zh) 2021-06-29 2021-06-29 水溶性蜂胶纳米颗粒的制备方法

Country Status (1)

Country Link
CN (1) CN113332453B (zh)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115536724B (zh) * 2022-09-30 2023-10-20 中国人民解放军陆军军医大学 水溶性锌(ii)-齐墩果酸荧光纳米颗粒的制备方法

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1596711A (zh) * 2004-08-23 2005-03-23 王美岭 蜂胶蜂王浆保健食品及其生产方法
CN1303899C (zh) * 2005-05-08 2007-03-14 浙江大学 纳米蜂胶制品的制备方法
CN101390884B (zh) * 2008-10-23 2011-05-11 陈宜斗 无醇制取天然蜂胶的方法
GB201001412D0 (en) * 2010-01-28 2010-03-17 Natures Lab Ltd Propolis and process for the treatment thereof
BRPI1004808B1 (pt) * 2010-10-27 2020-09-29 Embrapa - Empresa Brasileira De Pesquisa Agropecuária Composições baseadas em nanopartículas de própolis, processos de obtenção e uso
BR102015033031A2 (pt) * 2015-12-30 2017-07-04 Instituto De Tecnologia E Pesquisa Process of obtaining and composition of nanoencapsulates of red propolis extract
CN110604747B (zh) * 2019-10-30 2021-09-28 中国医学科学院药用植物研究所 一种蜂胶纳米粒、制备方法、应用及其冻干粉的制备方法

Also Published As

Publication number Publication date
CN113332453A (zh) 2021-09-03

Similar Documents

Publication Publication Date Title
WO2004098570B1 (en) Nanoparticulate bioactive agents
CN113332453B (zh) 水溶性蜂胶纳米颗粒的制备方法
CN109793720B (zh) 一种基于透明质酸的线粒体靶向抗氧化剂Mito Q纳米制剂的制备方法及应用
CN112999161A (zh) 一种杜仲京尼平苷酸复合纳米颗粒的制备方法
CN111568875A (zh) 具有核壳结构的纳米硒/介孔二氧化硅及其制备方法与应用
CN113083176A (zh) 一种空心原花青素壁材微胶囊的制备方法及其应用
CN1931129A (zh) 一种可生物降解聚合物磁性载药微球的制备方法
CN110604747B (zh) 一种蜂胶纳米粒、制备方法、应用及其冻干粉的制备方法
CN101036785A (zh) 肝炎疫苗的壳聚糖纳米粒口服制剂
CN112386584A (zh) 一种细菌响应性兽用恩诺沙星复合纳米系统及其制备方法
CA2542049C (en) Finely particulate composite containing carbon compound encapsulated therein
CN102293753B (zh) 辅酶q10纳米混悬剂冻干组合物及其制备方法和应用
CN114642652B (zh) 一种岩藻多糖自组装载药纳米粒制备及应用
CN114948880B (zh) 一种咖啡酸苯乙酯纳米稳定缓释剂型的制备方法
CN113633786B (zh) 一种牛血清白蛋白-疏水改性壳聚糖纳米微囊及其制备方法
CN111252803B (zh) 一种方形AgCl纳米颗粒及其制备方法和应用
Assimopoulou et al. Synthesis and release studies of shikonin-containing microcapsules prepared by the solvent evaporation method
CN108096218B (zh) 一种载柴胡皂甙a的纳米颗粒及其制备方法
CN108451931B (zh) 一种pah/f聚电解质复合物纳米载体及其制备方法
CN110859763B (zh) 一种负载脂溶性活性成分的皮克林乳液及制备方法与化妆品
CN109453265B (zh) 聚氰基丙烯酸正丁酯包裹纳米锌绿茶缓释制剂及其应用
CN114304143A (zh) 一种农药传递体系及其制备方法
KR102604038B1 (ko) 트로폴론이 담지된 키토산 나노입자 및 이의 제조방법
CN115466618B (zh) 一种黑磷量子点水性分散体及其制备方法和应用
CN113750074B (zh) 一种新型酸响应离子液体微胶囊及其制备方法与应用

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant