CN118370744A - Preparation method and application of turmeric fiber preparation - Google Patents
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Abstract
本发明提供一种姜黄纤维制剂的制备方法及其用途,步骤包括:将姜黄纤维粉用去离子水溶解后过滤,通过醇沉法制备姜黄水溶性纤维;将姜黄水溶性纤维溶解于去离子水中,加入乙醇,调解pH值为4.0~5.0,搅拌后得到纤维基液;将负载有姜黄素的蛋白‑壳聚糖纳米复合物溶解于无水乙醇中得到复合姜黄素液;将姜黄素液滴加至搅拌状态中的纤维基液中,加入乙醇后搅拌、旋转蒸发去除乙醇后用去离子水定容至原体积,得到姜黄纤维制剂。本发明所制备得到的姜黄纤维制剂可以改善姜黄素的低溶解度和低生物利用度问题,并与水溶性纤维共奏调节肠道之效,以期为扩大姜黄素类物质的食疗保健与临床应用提供新方法。
The present invention provides a preparation method and use of a turmeric fiber preparation, the steps include: filtering after dissolving turmeric fiber powder with deionized water, preparing turmeric water-soluble fiber by alcohol precipitation method; dissolving turmeric water-soluble fiber in deionized water, adding ethanol, adjusting pH value to 4.0-5.0, and stirring to obtain fiber-based liquid; dissolving the protein-chitosan nanocomposite loaded with curcumin in anhydrous ethanol to obtain a composite curcumin liquid; adding the curcumin liquid dropwise to the fiber-based liquid in a stirring state, stirring after adding ethanol, and removing ethanol by rotary evaporation, and then settling to the original volume with deionized water to obtain a turmeric fiber preparation. The turmeric fiber preparation prepared by the present invention can improve the low solubility and low bioavailability problems of curcumin, and play the effect of regulating the intestine with water-soluble fiber, in order to provide a new method for expanding the dietary health care and clinical application of curcumin-like substances.
Description
技术领域Technical Field
本发明涉及保健品技术领域,具体涉及一种姜黄纤维制剂的制备方法及其用途。The invention relates to the technical field of health products, and in particular to a preparation method and application of a turmeric fiber preparation.
背景技术Background technique
姜科植物姜黄(Curcuma longa L.)一直是亚洲文化和烹饪中不可或缺的一部分。姜黄素类是从姜黄根茎中提取的植物多酚类物质,主要以姜黄素、去甲氧基姜黄素和双去甲氧基姜黄素为代表。现代药理研究表明,姜黄素具有抗氧化、抗炎、抗肿瘤、抗增殖和抗菌等多种生物活性,在多种疾病的预防和治疗中效果甚佳。目前,大量关于姜黄素抗肿瘤活性的临床试验已经广泛开展,主要集中于针对胰腺癌和结肠癌。溃疡性结肠炎(Ulcerativecolitis,UC)是一种慢性特发性结肠炎症性肠病(Inflammatory bowel disease,IBD),该病以直肠、(乙状)结肠黏膜及黏膜下层的炎症和溃疡性病变为主,严重时可侵犯到全结肠和末端回肠,具有难治性、反复性、连续性、复发率高,聚集性及地域性等特点,需要长期监控和治疗,已被世界卫生组织(World Health Organization,WHO)列为现代难治疾病之一。研究显示合理饮食有助于缓解UC,这是由于饮食中的功能组分可以调节肠道菌群、修复黏膜屏障和增强免疫功能。尽管如此,通过常规饮食缓解UC的实际效果受患者的饮食喜好、饮食习惯以及食物来源的影响较大。因此,通过直接补充功能性组分缓解UC更符合现实需求。大量研究表明特定的组分如纤维、多酚、功能油脂等对UC具有明显的缓解作用。姜黄素的主要活性成分为姜黄素,姜黄素是常见的多酚,具有抗炎、抗氧化、抗肿瘤等生物活性,在临床研究中对UC的治疗、缓解和减少复发表现出了积极的效果。此外,姜黄作为烹饪食物必不可少的香料和染料,临床安全性评价研究表明姜黄素在日服计量高达8g至12g的大剂量下,依然没有造成任何明显的毒副作用。由此可知,姜黄素类是具有多种健康益处且安全性高的植物多酚类物质。Turmeric (Curcuma longa L.), a plant of the ginger family, has always been an integral part of Asian culture and cooking. Curcuminoids are plant polyphenols extracted from the rhizomes of turmeric, mainly represented by curcumin, demethoxycurcumin and bisdemethoxycurcumin. Modern pharmacological studies have shown that curcumin has multiple biological activities such as antioxidant, anti-inflammatory, anti-tumor, anti-proliferative and antibacterial, and is very effective in the prevention and treatment of a variety of diseases. At present, a large number of clinical trials on the anti-tumor activity of curcumin have been widely carried out, mainly focusing on pancreatic cancer and colon cancer. Ulcerative colitis (UC) is a chronic idiopathic inflammatory bowel disease (IBD) of the colon. The disease is mainly characterized by inflammation and ulcerative lesions in the rectal and (sigmoid) colon mucosa and submucosa. In severe cases, it can invade the entire colon and terminal ileum. It is characterized by refractory, recurrent, continuous, high recurrence rate, aggregation and regionality. It requires long-term monitoring and treatment. It has been listed as one of the modern refractory diseases by the World Health Organization (WHO). Studies have shown that a reasonable diet can help relieve UC, because the functional components in the diet can regulate intestinal flora, repair mucosal barriers and enhance immune function. Nevertheless, the actual effect of relieving UC through a regular diet is greatly affected by the patient's dietary preferences, dietary habits and food sources. Therefore, relieving UC by directly supplementing functional components is more in line with practical needs. A large number of studies have shown that specific components such as fiber, polyphenols, functional oils, etc. have a significant effect on UC. The main active ingredient of curcumin is curcumin, which is a common polyphenol with anti-inflammatory, antioxidant, and anti-tumor biological activities. In clinical studies, it has shown positive effects on the treatment, relief, and reduction of recurrence of UC. In addition, turmeric is an essential spice and dye for cooking food. Clinical safety evaluation studies have shown that curcumin does not cause any obvious toxic side effects at a daily dose of up to 8g to 12g. It can be seen that curcumin is a plant polyphenol with multiple health benefits and high safety.
然而,姜黄素因其高的疏水性和低的生物利用度,阻碍了其抗炎、抗氧化、抗肿瘤等生物活性的发挥;同时,低溶解度和低生物利用度也一直是限制姜黄素扩大开发和运用的难点和关键问题。市场上的姜黄素饮品常见于通过添加大量或者多种表面活性剂提高姜黄素的溶解性,提高饮品中姜黄素含量。该方法并不能良好的解决生物利用度的问题,还会在饮品中引入大量的表面活性剂,不利于饮品的安全性和存储稳定性。此外,为了掩盖姜黄素的辛、苦、辣的特殊味道,许多饮品都使用了很多的调味添加剂,这不仅可能影响饮品的口感和品质,还可能引入新的安全风险。However, due to its high hydrophobicity and low bioavailability, curcumin's anti-inflammatory, antioxidant, and anti-tumor biological activities are hindered; at the same time, low solubility and low bioavailability have always been the difficulties and key issues that limit the expansion of development and application of curcumin. Curcumin drinks on the market are often made by adding a large amount or multiple surfactants to increase the solubility of curcumin and increase the curcumin content in the drink. This method does not solve the problem of bioavailability well, and it will also introduce a large amount of surfactants into the drink, which is not conducive to the safety and storage stability of the drink. In addition, in order to cover up the special taste of curcumin, which is pungent, bitter, and spicy, many drinks use a lot of flavoring additives, which may not only affect the taste and quality of the drink, but also introduce new safety risks.
近年来,随着纤维制剂技术的发展,其在药物制剂领域的应用越来越广泛。纤维制剂具有比表面积大、吸附能力强、生物相容性好等优点,可以提高药物的溶解度和生物利用度,改善药物的稳定性和安全性。因此,将姜黄与纤维制剂技术相结合,开发一种新型的姜黄纤维制剂,具有重要的研究价值和实际应用前景。基于此,本发明提供了一种姜黄纤维制剂的制备方法及其用途。In recent years, with the development of fiber preparation technology, its application in the field of pharmaceutical preparations has become more and more extensive. Fiber preparations have the advantages of large specific surface area, strong adsorption capacity, good biocompatibility, etc., which can improve the solubility and bioavailability of drugs and improve the stability and safety of drugs. Therefore, combining turmeric with fiber preparation technology to develop a new type of turmeric fiber preparation has important research value and practical application prospects. Based on this, the present invention provides a preparation method of turmeric fiber preparation and its use.
发明内容Summary of the invention
本发明的目的是提供一种姜黄纤维制剂的制备方法及其用途,解决上述姜黄素类物质口服的低溶解度和低生物利用度问题,为姜黄素的食疗保健开发和临床应用提供新思路。The purpose of the present invention is to provide a preparation method of a turmeric fiber preparation and its use, so as to solve the problems of low solubility and low bioavailability of the above-mentioned curcuminoids when taken orally, and to provide new ideas for the development of dietary health care and clinical application of curcumin.
一方面,本发明提供一种姜黄纤维制剂,步骤包括:In one aspect, the present invention provides a turmeric fiber preparation, comprising the steps of:
(1)制备姜黄水溶性纤维:将姜黄纤维粉用去离子水溶解后过滤,通过醇沉法制备姜黄水溶性纤维;(1) Preparing turmeric water-soluble fiber: Dissolving turmeric fiber powder in deionized water and filtering, and preparing turmeric water-soluble fiber by alcohol precipitation method;
(2)制备纤维基液和姜黄素液:将姜黄水溶性纤维溶解于去离子水中,加入乙醇,调解pH值为4.0~5.0,搅拌后得到纤维基液;将负载有姜黄素的蛋白-壳聚糖纳米复合物溶解于无水乙醇中得到复合姜黄素液;(2) preparing a fiber-based liquid and a curcumin liquid: dissolving turmeric water-soluble fiber in deionized water, adding ethanol, adjusting the pH value to 4.0-5.0, and stirring to obtain a fiber-based liquid; dissolving a protein-chitosan nanocomposite loaded with curcumin in anhydrous ethanol to obtain a composite curcumin liquid;
(3)制备姜黄纤维制剂:将复合姜黄素液滴加至搅拌状态中的纤维基液中,加入乙醇后搅拌、旋转蒸发去除乙醇后用去离子水定容至原体积,得到姜黄纤维制剂。(3) Preparation of turmeric fiber preparation: adding the composite curcumin solution dropwise to the fiber base solution in a stirring state, adding ethanol and stirring, removing the ethanol by rotary evaporation, and then fixing the volume to the original volume with deionized water to obtain the turmeric fiber preparation.
进一步地,所述姜黄纤维粉和去离子水料液比为1:(25-35),所述去离子水的温度为65-75℃,溶解在搅拌条件下进行,搅拌时间为25-35min。Furthermore, the material-liquid ratio of the turmeric fiber powder to deionized water is 1:(25-35), the temperature of the deionized water is 65-75° C., and the dissolution is carried out under stirring conditions for 25-35 minutes.
进一步地,所述醇沉法的具体方法包括:将所得滤液与3-5倍体积的乙醇混合并静置0.8-1.2天,于7500-8500rpm离心12-18min,随后将沉淀冷冻干燥。Furthermore, the specific method of the alcohol precipitation method includes: mixing the obtained filtrate with 3-5 times the volume of ethanol and standing for 0.8-1.2 days, centrifuging at 7500-8500 rpm for 12-18 minutes, and then freeze-drying the precipitate.
进一步地,所述步骤(2)中加入乙醇使得含量达到35-40vol%,所述调解pH的试剂为0.2M HCl和NaOH溶液。Furthermore, in the step (2), ethanol is added to a content of 35-40 vol%, and the reagents for adjusting pH are 0.2 M HCl and NaOH solutions.
进一步地,所述负载有姜黄素的蛋白-壳聚糖纳米复合物的制备方法包括:将乳铁蛋白和壳聚糖分别溶解在两份解离溶液中;将姜黄素溶解在乙醇中,将pH值调至5.5,并在3-5℃下孵育10h以上以完全解离;将姜黄素乙醇溶液与乳铁蛋白溶液混合得到混合液一,随后将混溶液一和壳聚糖溶液等比例混合,透析去除尿素;完成后得到混合液二,离心收集上清液即得。Furthermore, the preparation method of the protein-chitosan nanocomposite loaded with curcumin includes: dissolving lactoferrin and chitosan in two dissociation solutions respectively; dissolving curcumin in ethanol, adjusting the pH value to 5.5, and incubating at 3-5°C for more than 10 hours for complete dissociation; mixing the curcumin ethanol solution with the lactoferrin solution to obtain a mixed solution 1, then mixing the mixed solution 1 and the chitosan solution in equal proportions, and dialyzing to remove urea; after completion, a mixed solution 2 is obtained, and the supernatant is collected by centrifugation.
进一步地,所述壳聚糖Mw=200ku,脱乙酰度≥90%;所述解离溶液中包括0.5%冰醋酸,6mol/L尿素;所述乳铁蛋白与解离溶液的用量比为(200-220)mg:(24-30)mL;所述壳聚糖与解离溶液的用量比为(200-220)mg:(24-30)mL;所述姜黄素与乙醇的用量比为(500-600)mg:(18-22)mL。Furthermore, the chitosan has a Mw of 200 ku and a deacetylation degree of ≥90%; the dissociation solution comprises 0.5% glacial acetic acid and 6 mol/L urea; the dosage ratio of lactoferrin to the dissociation solution is (200-220) mg: (24-30) mL; the dosage ratio of chitosan to the dissociation solution is (200-220) mg: (24-30) mL; and the dosage ratio of curcumin to ethanol is (500-600) mg: (18-22) mL.
进一步地,所述pH值调节剂为乙酸。Furthermore, the pH adjuster is acetic acid.
进一步地,所述透析材料为分子质量为8.0~14ku的透析管;透析方式为在2-3℃下用去离子水透析2-2.5天以去除尿素。Furthermore, the dialysis material is a dialysis tube with a molecular weight of 8.0-14 ku; the dialysis method is to use deionized water for 2-2.5 days at 2-3°C to remove urea.
进一步地,所述离心条件为8 000g、15-25min。Furthermore, the centrifugation conditions are 8 000 g, 15-25 min.
进一步地,所述复合姜黄素液中的姜黄素与姜黄水溶性纤维的质量比为1:20-25。Furthermore, the mass ratio of curcumin to turmeric water-soluble fiber in the composite curcumin liquid is 1:20-25.
进一步地,所述复合姜黄素液的浓度为3-7mg/mL。Furthermore, the concentration of the compound curcumin solution is 3-7 mg/mL.
进一步地,所述姜黄素纯度>95%,每10mg姜黄素包括姜黄素72.22wt%、去甲氧基姜黄素17.72wt%和双去甲氧基姜黄素5.26wt%。Furthermore, the curcumin has a purity of >95%, and each 10 mg of curcumin comprises 72.22 wt % of curcumin, 17.72 wt % of demethoxycurcumin and 5.26 wt % of bisdemethoxycurcumin.
进一步地,所述步骤(3)中加入乙醇后乙醇含量达到35-40vol%,所述搅拌为在室温下于550-650rpm继续搅拌2-4h。Furthermore, after adding ethanol in step (3), the ethanol content reaches 35-40 vol%, and the stirring is continued at 550-650 rpm for 2-4 hours at room temperature.
进一步地,所述旋转蒸发的条件为置于38-45℃水浴中,旋转蒸发的时间为17-23℃。Furthermore, the rotary evaporation condition is placed in a 38-45°C water bath, and the rotary evaporation time is 17-23°C.
另一方面,本发明中姜黄纤维制剂应用于制备保健品或食品中。On the other hand, the turmeric fiber preparation of the present invention is used in preparing health products or foods.
有益效果:Beneficial effects:
本发明提供的姜黄纤维制剂能够防治UC,利用多酚类物质与纤维的天然相互作用力,选取姜黄自体的水溶性纤维对难溶性的姜黄素进行包载,克服了纤维与姜黄素之间溶解度的差异,构建了共同溶解体系(40%乙醇),以促进二者之间的化学相互作用,并通过旋转蒸发除去体系中的乙醇,能够辅助姜黄素的包载,有助于包装组装纤维纳米粒的形成。The turmeric fiber preparation provided by the present invention can prevent and treat UC, utilizes the natural interaction force between polyphenols and fibers, selects water-soluble fibers of turmeric itself to encapsulate poorly soluble curcumin, overcomes the difference in solubility between the fibers and curcumin, constructs a co-dissolving system (40% ethanol) to promote the chemical interaction between the two, and removes ethanol in the system by rotary evaporation, which can assist the encapsulation of curcumin and facilitates the formation of packaged and assembled fiber nanoparticles.
负载有姜黄素的蛋白-壳聚糖纳米复合物在制备姜黄纤维制剂时对结肠组织氧化应激的缓解能力较好,通过纳米复合物的负载,姜黄素的溶解度得到提高,从而使其在体内的吸收和分布更加均匀,提高了其在结肠组织中的浓度,这也有效提高了姜黄素的生物利用度;蛋白-壳聚糖纳米复合物具有良好的生物相容性和靶向性,可以实现对结肠组织的靶向输送,使姜黄素更多地集中在结肠部位,发挥其抗氧化作用;此外,纳米复合物可以实现对姜黄素的缓释,使药物在结肠部位持续释放,从而维持较高的药物浓度,长时间地缓解结肠组织的氧化应激反应。同时,蛋白和壳聚糖本身也可能具有一定的抗氧化和抗炎作用,与姜黄素共同发挥作用,进一步增强对结肠组织氧化应激的缓解能力。The protein-chitosan nanocomposite loaded with curcumin has a good ability to relieve the oxidative stress of colon tissue when preparing turmeric fiber preparations. Through the loading of the nanocomposite, the solubility of curcumin is improved, so that its absorption and distribution in the body are more uniform, and its concentration in the colon tissue is increased, which also effectively improves the bioavailability of curcumin; the protein-chitosan nanocomposite has good biocompatibility and targeting, which can achieve targeted delivery to colon tissue, so that curcumin is more concentrated in the colon and exerts its antioxidant effect; in addition, the nanocomposite can achieve sustained release of curcumin, so that the drug is continuously released in the colon, thereby maintaining a high drug concentration and relieving the oxidative stress response of colon tissue for a long time. At the same time, protein and chitosan themselves may also have certain antioxidant and anti-inflammatory effects, and work together with curcumin to further enhance the ability to relieve oxidative stress in colon tissue.
纤维具有调节胃肠道功能、降血糖等多种健康益处,其亦能通过作用于肠道菌群从而促进姜黄素类物质的吸收和利用。因此,本发明在提高姜黄素生物利用度的同时,还得到一种新兴的功能食品组分即“防治UC纤维”,从而丰富姜黄素类补充剂品种、促进保健食品产业发展。Fiber has many health benefits such as regulating gastrointestinal function and lowering blood sugar, and it can also promote the absorption and utilization of curcuminoids by acting on intestinal flora. Therefore, the present invention not only improves the bioavailability of curcumin, but also obtains a new functional food component, namely "UC prevention and treatment fiber", thereby enriching the variety of curcuminoid supplements and promoting the development of the health food industry.
附图说明BRIEF DESCRIPTION OF THE DRAWINGS
图1为本发明实验例1中DAI分数曲线图;FIG1 is a DAI score curve diagram in Experimental Example 1 of the present invention;
图2为本发明实验例1中结肠形态得分对应图;FIG2 is a graph showing the colon morphology scores in Experimental Example 1 of the present invention;
图3为本发明实验例1中结肠形态HE染色图;FIG3 is a HE staining diagram of the colon morphology in Experimental Example 1 of the present invention;
图4为本发明实验例1中D-乳酸浓度图;FIG4 is a graph showing the concentration of D-lactic acid in Experimental Example 1 of the present invention;
图5为本发明实验例1中GSH浓度图;FIG5 is a graph showing the concentration of GSH in Experimental Example 1 of the present invention;
图6为本发明实验例1中SOD浓度图;FIG6 is a graph showing the concentration of SOD in Experimental Example 1 of the present invention;
图7为本发明实验例1中IL-1β浓度图;FIG7 is a graph showing IL-1β concentration in Experimental Example 1 of the present invention;
图8为本发明实验例1中IL-6浓度图;FIG8 is a graph showing IL-6 concentration in Experimental Example 1 of the present invention;
图9为本发明实验例1中IL-10浓度图;FIG9 is a graph showing IL-10 concentration in Experimental Example 1 of the present invention;
图10为本发明实验例1中TNF-α浓度图。FIG. 10 is a graph showing the concentration of TNF-α in Experimental Example 1 of the present invention.
具体实施方式Detailed ways
下面对本发明的技术方案进行清楚、完整地描述,显然,所描述的实施例是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。The technical solution of the present invention is described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
下面将结合实施例对本发明的实施方案进行详细描述,但是本领域技术人员将会理解,下列实施例仅用于说明本发明,而不应视为限制本发明的范围,实施例中未注明的具体条件,按照常规条件或者制造商建议的条件进行,所用试剂或仪器未注明生产厂商者,均为可以通过市售购买获得的常规产品。The embodiments of the present invention will be described in detail below in conjunction with examples. However, those skilled in the art will understand that the following examples are only used to illustrate the present invention and should not be construed as limiting the scope of the present invention. The specific conditions not specified in the examples are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used without indicating the manufacturer are all conventional products that can be purchased commercially.
姜黄纤维粉购自陕西绿莱生物科技有限公司,产品批号:LL211012-1。Turmeric fiber powder was purchased from Shaanxi Lvlai Biotechnology Co., Ltd., product batch number: LL211012-1.
实施例1Example 1
本实施例提供一种姜黄纤维制剂的制备方法,步骤包括:This embodiment provides a method for preparing a turmeric fiber preparation, the steps comprising:
(1)负载有姜黄素的蛋白-壳聚糖纳米复合物的制备:将210mg乳铁蛋白和210mgMw=200ku,脱乙酰度≥90%的壳聚糖分别溶解在两份27mL含有0.5%冰醋酸,6mol/L尿素的解离溶液中;用乙酸将最终pH值调至5.5,并在4℃下孵育12h以完全解离;将550mg姜黄素溶解在20mL乙醇中得到姜黄素乙醇溶液;将姜黄素乙醇溶液与乳铁蛋白溶液混合得到混合液一,将混溶液一和壳聚糖溶液等比例混合,使用分子质量为10ku的透析管在2.5℃下用去离子水透析2天以去除尿素;透析完成后得到混合液二,在8 000g条件下离心20min,收集上清液即得;(1) Preparation of protein-chitosan nanocomposites loaded with curcumin: 210 mg lactoferrin and 210 mg chitosan with a Mw=200 ku and a deacetylation degree of ≥90% were dissolved in two 27 mL dissociation solutions containing 0.5% glacial acetic acid and 6 mol/L urea, respectively; the final pH value was adjusted to 5.5 with acetic acid, and the mixture was incubated at 4°C for 12 h for complete dissociation; 550 mg curcumin was dissolved in 20 mL ethanol to obtain a curcumin ethanol solution; the curcumin ethanol solution was mixed with the lactoferrin solution to obtain a mixed solution 1; the mixed solution 1 and the chitosan solution were mixed in equal proportions, and dialyzed with deionized water at 2.5°C for 2 days using a dialysis tube with a molecular weight of 10 ku to remove urea; after the dialysis was completed, a mixed solution 2 was obtained, which was centrifuged at 8,000 g for 20 min, and the supernatant was collected;
其中,10mg姜黄素(纯度>95%)包括姜黄素72.22wt%、去甲氧基姜黄素17.72wt%和双去甲氧基姜黄素5.26wt%。Among them, 10 mg of curcumin (purity>95%) includes 72.22 wt % of curcumin, 17.72 wt % of demethoxycurcumin and 5.26 wt % of bisdemethoxycurcumin.
(2)复合姜黄素液的制备:将负载有姜黄素的蛋白-壳聚糖纳米复合物溶解于无水乙醇中得到复合姜黄素液,复合姜黄素液的浓度为5mg/mL;(2) Preparation of composite curcumin solution: dissolving the protein-chitosan nanocomposite loaded with curcumin in anhydrous ethanol to obtain a composite curcumin solution, wherein the concentration of the composite curcumin solution is 5 mg/mL;
(3)姜黄水溶性纤维的制备:将姜黄纤维粉置于70℃水中,料液比为1:30,搅拌30分钟后,过滤;将所得滤液与4倍体积的乙醇混合并静置1天,于8000rpm离心15min,将沉淀冷冻干燥后,得到姜黄水溶性纤维;(3) Preparation of turmeric water-soluble fiber: turmeric fiber powder was placed in 70°C water with a solid-liquid ratio of 1:30, stirred for 30 minutes, and filtered; the obtained filtrate was mixed with 4 volumes of ethanol and allowed to stand for 1 day, centrifuged at 8000 rpm for 15 minutes, and the precipitate was freeze-dried to obtain turmeric water-soluble fiber;
(4)纤维基液的制备:将制备的姜黄水溶性纤维溶解于纯水中,加入乙醇,使乙醇含量达到40vol%,采用0.2M HCl和NaOH溶液调节pH值至4,搅拌,得到纤维基液;(4) Preparation of fiber-based liquid: dissolving the prepared turmeric water-soluble fiber in pure water, adding ethanol to make the ethanol content reach 40 vol%, adjusting the pH value to 4 with 0.2 M HCl and NaOH solution, stirring, and obtaining a fiber-based liquid;
其中,复合姜黄素液中的姜黄素与姜黄水溶性纤维的质量比为1:30;Among them, the mass ratio of curcumin to turmeric water-soluble fiber in the composite curcumin solution is 1:30;
(5)将复合姜黄素液滴加至搅拌状态中的纤维基液中,再滴加乙醇使乙醇含量达到40vol%,室温下于600rpm继续搅拌3h后,置于40℃水浴上旋转蒸发20min除去乙醇,用水定容至原体积,得到姜黄纤维制剂。(5) The composite curcumin liquid was added dropwise to the fiber base liquid in a stirring state, and ethanol was then added dropwise to make the ethanol content reach 40 vol%, and the mixture was stirred at 600 rpm for 3 h at room temperature, and then placed in a 40° C. water bath for 20 min to remove the ethanol by rotary evaporation, and the volume was adjusted to the original volume with water to obtain a turmeric fiber preparation.
对比例1Comparative Example 1
本对比例制备复合姜黄素的步骤包括:The steps of preparing the compound curcumin in this comparative example include:
(1)负载有姜黄素的蛋白-壳聚糖纳米复合物的制备:将210mg乳铁蛋白和210mgMw=200ku,脱乙酰度≥90%的壳聚糖分别溶解在两份27mL含有0.5%冰醋酸,6mol/L尿素的解离溶液中;用乙酸将最终pH值调至5.5,并在4℃下孵育12h以完全解离;将550mg姜黄素溶解在20mL乙醇中得到姜黄素乙醇溶液;将姜黄素乙醇溶液与乳铁蛋白溶液混合得到混合液一,将混溶液一和壳聚糖溶液等比例混合,使用分子质量为10ku的透析管在2.5℃下用去离子水透析2天以去除尿素;透析完成后得到混合液二,在8 000g条件下离心20min,收集上清液即得;(1) Preparation of protein-chitosan nanocomposites loaded with curcumin: 210 mg lactoferrin and 210 mg chitosan with a Mw=200 ku and a deacetylation degree of ≥90% were dissolved in two 27 mL dissociation solutions containing 0.5% glacial acetic acid and 6 mol/L urea, respectively; the final pH value was adjusted to 5.5 with acetic acid, and the mixture was incubated at 4°C for 12 h for complete dissociation; 550 mg curcumin was dissolved in 20 mL ethanol to obtain a curcumin ethanol solution; the curcumin ethanol solution was mixed with the lactoferrin solution to obtain a mixed solution 1; the mixed solution 1 and the chitosan solution were mixed in equal proportions, and dialyzed with deionized water at 2.5°C for 2 days using a dialysis tube with a molecular weight of 10 ku to remove urea; after the dialysis was completed, a mixed solution 2 was obtained, which was centrifuged at 8,000 g for 20 min, and the supernatant was collected;
其中,10mg姜黄素(纯度>95%)包括姜黄素72.22wt%、去甲氧基姜黄素17.72wt%和双去甲氧基姜黄素5.26wt%。Among them, 10 mg of curcumin (purity>95%) includes 72.22 wt % of curcumin, 17.72 wt % of demethoxycurcumin and 5.26 wt % of bisdemethoxycurcumin.
(2)复合姜黄素液的制备:将负载有姜黄素的蛋白-壳聚糖纳米复合物溶解于无水乙醇中得到复合姜黄素液,复合姜黄素液的浓度为5mg/mL。(2) Preparation of composite curcumin solution: The protein-chitosan nanocomposite loaded with curcumin was dissolved in anhydrous ethanol to obtain a composite curcumin solution, wherein the concentration of the composite curcumin solution was 5 mg/mL.
对比例2Comparative Example 2
本对比例制备姜黄水溶性纤维的步骤包括:将姜黄纤维粉置于70℃水中,料液比为1:30,搅拌30分钟后,过滤;将所得滤液与4倍体积的乙醇混合并静置1天,于8000rpm离心15min,将沉淀冷冻干燥后,得到姜黄水溶性纤维。The steps of preparing turmeric water-soluble fiber in this comparative example include: placing turmeric fiber powder in 70° C. water with a solid-liquid ratio of 1:30, stirring for 30 minutes, and filtering; mixing the obtained filtrate with 4 times the volume of ethanol and standing for 1 day, centrifuging at 8000 rpm for 15 minutes, and freeze-drying the precipitate to obtain turmeric water-soluble fiber.
为了进一步说明本发明提供的防治UC纤维制剂的性能和有益效果,特进行下述实验:In order to further illustrate the performance and beneficial effects of the fiber preparation for preventing and treating UC provided by the present invention, the following experiments were conducted:
试验例1Test Example 1
通过自由给予饮用3% DSS进行急性UC造模,美莎拉嗪(mesalazine,5-ASA)作为阳性对照药物。将48只雄性SPF级BALB/c小鼠适应性喂养7天后随机分为六组:对照组、模型组、阳性药组、复合姜黄素组(对比例1制备)、姜黄水溶性纤维组(对比例2制备)及姜黄纤维制剂组(实施例1制备)。第1天开始,除对照组小鼠外,其余各组灌胃给予相应受试药,1次/日,并自由饮用3% DSS溶液诱导UC模型,连续9天;对照组每天灌胃和饮用无菌去离子水。实验过程中每天称量小鼠质量,进行疾病活动指数(disease active index,DAI)评分。实验第10天,摘眼球采血后处死并解剖小鼠,取结肠肠段测量长度并进行组织学损伤评分,取部分结肠固定于4%多聚甲醛溶液中,石蜡包埋切片,苏木精-伊红染色(Hematoxylin andeosin staining,HE)观察结肠黏膜病理变化;采用生化分析法检测结肠组织超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽(glutathione,GSH);同时采用酶联免疫吸附法测定血浆中白细胞介素-1β(interleukin-β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-10(interleukin-10,IL-10)及肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)含量;微量法检测D-乳酸(D(-)-lactic acid)含量。结果见下表:Acute UC modeling was performed by freely giving 3% DSS to drink, and mesalazine (5-ASA) was used as a positive control drug. After 7 days of adaptive feeding, 48 male SPF-grade BALB/c mice were randomly divided into six groups: control group, model group, positive drug group, compound curcumin group (prepared in comparative example 1), turmeric water-soluble fiber group (prepared in comparative example 2) and turmeric fiber preparation group (prepared in Example 1). Starting from the first day, except for the control group mice, the remaining groups were gavaged with the corresponding test drugs, once a day, and freely drank 3% DSS solution to induce UC model for 9 consecutive days; the control group was gavaged and drank sterile deionized water every day. The mice were weighed every day during the experiment, and the disease activity index (DAI) was scored. On the 10th day of the experiment, the mice were killed and blood was collected after eyeball removal. The length of the colon segment was measured and histological damage was scored. Part of the colon was fixed in 4% paraformaldehyde solution, embedded in paraffin and sectioned. Hematoxylin andeosin staining (HE) was used to observe the pathological changes of colon mucosa; biochemical analysis was used to detect superoxide dismutase (SOD) and glutathione (GSH) in colon tissue; enzyme-linked immunosorbent assay was used to determine the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) in plasma; the content of D-lactic acid (D(-)-lactic acid) was detected by microassay. The results are shown in the following table:
表1DAI得分表(平均数±标准差,n=8)Table 1 DAI score table (mean ± standard deviation, n = 8)
表2结肠组织肉眼大体形态学评分(平均数±标准差,n=8)Table 2 Macroscopic morphological scores of colon tissue (mean ± SD, n = 8)
图1为本发明实验例1中DAI分数曲线图;图2为本发明实验例1中结肠形态得分对应图;图3为本发明实验例1中结肠形态HE染色图;图4为本发明实验例1中D-乳酸浓度图;图5为本发明实验例1中GSH浓度图;图6为本发明实验例1中SOD浓度图;图7为本发明实验例1中IL-1β浓度图;FIG. 1 is a graph of DAI scores in Experimental Example 1 of the present invention; FIG. 2 is a graph corresponding to colon morphology scores in Experimental Example 1 of the present invention; FIG. 3 is a HE staining graph of colon morphology in Experimental Example 1 of the present invention; FIG. 4 is a graph of D-lactic acid concentration in Experimental Example 1 of the present invention; FIG. 5 is a graph of GSH concentration in Experimental Example 1 of the present invention; FIG. 6 is a graph of SOD concentration in Experimental Example 1 of the present invention; and FIG. 7 is a graph of IL-1β concentration in Experimental Example 1 of the present invention;
图8为本发明实验例1中IL-6浓度图;图9为本发明实验例1中IL-10浓度图;图10为本发明实验例1中TNF-α浓度图。FIG. 8 is a graph showing the concentration of IL-6 in Experimental Example 1 of the present invention; FIG. 9 is a graph showing the concentration of IL-10 in Experimental Example 1 of the present invention; and FIG. 10 is a graph showing the concentration of TNF-α in Experimental Example 1 of the present invention.
由表1和图1可得,姜黄纤维制剂组的DAI评分低于模型组、复合姜黄素组和姜黄水溶性纤维组,DAI评分可判断结肠炎症状的严重程度,可见姜黄纤维制剂缓解结肠炎症效果较强;由表2和图2可得姜黄纤维制剂组的结肠形态评分低于模型组、复合姜黄素组和姜黄水溶性纤维组,结肠形态可直接判断结肠炎症状的严重程度,可见姜黄纤维制剂对结肠形态的保护作用较好;As shown in Table 1 and Figure 1, the DAI score of the turmeric fiber preparation group was lower than that of the model group, the compound curcumin group and the turmeric water-soluble fiber group. The DAI score can judge the severity of colitis symptoms, which shows that the turmeric fiber preparation has a strong effect in relieving colon inflammation; As shown in Table 2 and Figure 2, the colon morphology score of the turmeric fiber preparation group was lower than that of the model group, the compound curcumin group and the turmeric water-soluble fiber group. The colon morphology can directly judge the severity of colitis symptoms, which shows that the turmeric fiber preparation has a good protective effect on colon morphology;
由图3可得,与对照组相比,姜黄纤维制剂组与阳性药组中组织病理学结构以及中性粒细胞的浸润没有显著变化,而杯状细胞的数量显著增加且排列有序,模型组、复合姜黄素组和姜黄水溶性纤维组存在较多炎症浸润,肠壁结构损伤,可见姜黄纤维制剂对肠道具有保护作用,并减少DSS诱导的结肠组织损伤;As shown in Figure 3, compared with the control group, the histopathological structure and neutrophil infiltration in the turmeric fiber preparation group and the positive drug group did not change significantly, while the number of goblet cells increased significantly and was arranged in an orderly manner. There were more inflammatory infiltrations and intestinal wall structural damage in the model group, compound curcumin group and turmeric water-soluble fiber group. It can be seen that the turmeric fiber preparation has a protective effect on the intestine and reduces DSS-induced colon tissue damage;
由图4可得,姜黄纤维制剂组的D-乳酸含量低于模型组、复合姜黄素组和姜黄水溶性纤维组,当肠黏膜通透性增加时,肠道中细菌产生的大量D-乳酸通过受损黏膜入血,使血浆D-乳酸水平升高,可见姜黄纤维制剂对肠道具有保护作用;As shown in Figure 4, the D-lactic acid content in the turmeric fiber preparation group was lower than that in the model group, the compound curcumin group and the turmeric water-soluble fiber group. When the intestinal mucosal permeability increases, a large amount of D-lactic acid produced by bacteria in the intestine enters the blood through the damaged mucosa, increasing the plasma D-lactic acid level. It can be seen that the turmeric fiber preparation has a protective effect on the intestine.
由图5可得,姜黄纤维制剂组的GSH含量高于模型组、复合姜黄素组和姜黄水溶性纤维组,GSH能帮助机体保持正常的免疫系统功能,并有抗氧化等作用,可评估各组小鼠的氧化应激水平,可见姜黄纤维制剂对结肠组织氧化应激的缓解能力较好;As shown in Figure 5, the GSH content of the turmeric fiber preparation group was higher than that of the model group, the compound curcumin group and the turmeric water-soluble fiber group. GSH can help the body maintain normal immune system function and has antioxidant effects. It can be used to evaluate the oxidative stress level of mice in each group. It can be seen that the turmeric fiber preparation has a good ability to alleviate the oxidative stress of colon tissue.
由图6可得,姜黄纤维制剂组的SOD含量高于模型组、复合姜黄素组和姜黄水溶性纤维组,SOD是抗氧化酶,可以催化超氧化物阴离子自由基歧化生成氧气和过氧化氢,在体内氧化和抗氧化活性的平衡中发挥着至关重要的作用,可评估各组小鼠的氧化应激水平,可见姜黄纤维制剂对结肠组织氧化应激的缓解能力较好;As shown in Figure 6, the SOD content in the turmeric fiber preparation group was higher than that in the model group, the compound curcumin group and the turmeric water-soluble fiber group. SOD is an antioxidant enzyme that can catalyze the dismutation of superoxide anion radicals to generate oxygen and hydrogen peroxide. It plays a vital role in the balance of oxidation and antioxidant activity in the body and can evaluate the oxidative stress level of mice in each group. It can be seen that the turmeric fiber preparation has a good ability to alleviate the oxidative stress of colon tissue.
由图7可得,姜黄纤维制剂组的IL-1β含量低于模型组、复合姜黄素组和姜黄水溶性纤维组,IL-1β的大量释放直接导致肠道炎症的发展,可见姜黄纤维制剂对该炎症因子降低能力较好,缓解肠道炎症能力较好;As shown in Figure 7, the IL-1β content in the turmeric fiber preparation group was lower than that in the model group, the compound curcumin group and the turmeric water-soluble fiber group. The large release of IL-1β directly led to the development of intestinal inflammation. It can be seen that the turmeric fiber preparation has a better ability to reduce this inflammatory factor and relieve intestinal inflammation.
由图8可得,姜黄纤维制剂组的IL-6含量低于模型组、复合姜黄素组和姜黄水溶性纤维组,IL-6的大量释放直接导致肠道炎症的发展,可见姜黄纤维制剂对该炎症因子降低能力较好,缓解肠道炎症能力较好;As shown in Figure 8, the IL-6 content in the turmeric fiber preparation group was lower than that in the model group, the compound curcumin group and the turmeric water-soluble fiber group. The large release of IL-6 directly led to the development of intestinal inflammation. It can be seen that the turmeric fiber preparation has a better ability to reduce this inflammatory factor and relieve intestinal inflammation.
由图9可得,姜黄纤维制剂组的IL-10含量高于模型组、复合姜黄素组和姜黄水溶性纤维组,IL-10的缺乏会加剧结肠炎症的发展,可见姜黄纤维制剂对该抗炎因子增高能力较好,缓解肠道炎症能力较好;As shown in Figure 9, the IL-10 content of the turmeric fiber preparation group was higher than that of the model group, the compound curcumin group and the turmeric water-soluble fiber group. The lack of IL-10 would aggravate the development of colon inflammation. It can be seen that the turmeric fiber preparation has a better ability to increase this anti-inflammatory factor and relieve intestinal inflammation.
由图10可得,姜黄纤维制剂组的TNF-α含量低于模型组、复合姜黄素组和姜黄水溶性纤维组,TNF-α的大量释放直接导致肠道炎症的发展,可见姜黄纤维制剂对该炎症因子降低能力较好,缓解肠道炎症能力较好;As shown in Figure 10, the TNF-α content in the turmeric fiber preparation group was lower than that in the model group, the compound curcumin group and the turmeric water-soluble fiber group. The large release of TNF-α directly led to the development of intestinal inflammation. It can be seen that the turmeric fiber preparation has a better ability to reduce this inflammatory factor and relieve intestinal inflammation.
综上可知,与模型组相比,姜黄纤维制剂有效缓解了DAI评分水平、血液和组织炎症蛋白水平(TNF-α、IL-6、IL-10和IL-1β),并强于单一使用游离姜黄素和姜黄水溶性纤维;在HE染色下也体现姜黄纤维制剂对结肠形态的良好保持作用且有效下调血浆中D-乳酸水平;同时SOD与GSH含量高,与模型组相比,显著增加了对UC的抗氧化作用,并强于单一使用游离姜黄素和姜黄水溶性纤维。In summary, compared with the model group, the turmeric fiber preparation effectively alleviated the DAI score level, blood and tissue inflammatory protein levels (TNF-α, IL-6, IL-10 and IL-1β), and was stronger than the single use of free curcumin and turmeric water-soluble fiber; HE staining also showed that the turmeric fiber preparation had a good maintenance effect on the colon morphology and effectively lowered the level of D-lactic acid in plasma; at the same time, the SOD and GSH contents were high, and compared with the model group, it significantly increased the antioxidant effect on UC, and was stronger than the single use of free curcumin and turmeric water-soluble fiber.
实施例2Example 2
本实施例提供一种姜黄纤维制剂的制备方法,步骤包括:This embodiment provides a method for preparing a turmeric fiber preparation, the steps comprising:
(1)负载有姜黄素的蛋白-壳聚糖纳米复合物的制备:将200mg乳铁蛋白和210mgMw=200ku,脱乙酰度≥90%的壳聚糖分别溶解在两份24mL含有0.5%冰醋酸,6mol/L尿素的解离溶液中;用乙酸将最终pH值调至5.5,并在4℃下孵育10h以完全解离;将500mg姜黄素溶解在18mL乙醇中得到姜黄素乙醇溶液;将姜黄素乙醇溶液与乳铁蛋白溶液混合得到混合液一,将混溶液一和壳聚糖溶液等比例混合,使用分子质量为10ku的透析管在2.5℃下用去离子水透析2天以去除尿素;透析完成后得到混合液二,在8 000g条件下离心15min,收集上清液即得;(1) Preparation of protein-chitosan nanocomposites loaded with curcumin: 200 mg lactoferrin and 210 mg chitosan with Mw=200 ku and a deacetylation degree of ≥90% were dissolved in two 24 mL dissociation solutions containing 0.5% glacial acetic acid and 6 mol/L urea, respectively; the final pH value was adjusted to 5.5 with acetic acid, and the mixture was incubated at 4°C for 10 h for complete dissociation; 500 mg curcumin was dissolved in 18 mL ethanol to obtain a curcumin ethanol solution; the curcumin ethanol solution was mixed with the lactoferrin solution to obtain a mixed solution 1; the mixed solution 1 and the chitosan solution were mixed in equal proportions, and dialyzed with deionized water at 2.5°C for 2 days using a dialysis tube with a molecular weight of 10 ku to remove urea; after the dialysis was completed, a mixed solution 2 was obtained, which was centrifuged at 8,000 g for 15 min, and the supernatant was collected;
其中,10mg姜黄素(纯度>95%)包括姜黄素72.22wt%、去甲氧基姜黄素17.72wt%和双去甲氧基姜黄素5.26wt%。Among them, 10 mg of curcumin (purity>95%) includes 72.22 wt % of curcumin, 17.72 wt % of demethoxycurcumin and 5.26 wt % of bisdemethoxycurcumin.
(2)复合姜黄素液的制备:将负载有姜黄素的蛋白-壳聚糖纳米复合物溶解于无水乙醇中得到复合姜黄素液;复合姜黄素液的浓度为4mg/mL;(2) Preparation of composite curcumin solution: dissolving the protein-chitosan nanocomposite loaded with curcumin in anhydrous ethanol to obtain composite curcumin solution; the concentration of the composite curcumin solution is 4 mg/mL;
(3)姜黄水溶性纤维的制备:将姜黄纤维粉置于70℃水中,料液比为1:30,搅拌30分钟后,过滤;将所得滤液与4倍体积的乙醇混合并静置1天,于8000rpm离心15min,将沉淀冷冻干燥后,得到姜黄水溶性纤维;(3) Preparation of turmeric water-soluble fiber: turmeric fiber powder was placed in 70°C water with a solid-liquid ratio of 1:30, stirred for 30 minutes, and filtered; the obtained filtrate was mixed with 4 volumes of ethanol and allowed to stand for 1 day, centrifuged at 8000 rpm for 15 minutes, and the precipitate was freeze-dried to obtain turmeric water-soluble fiber;
(4)纤维基液的制备:将制备的姜黄水溶性纤维溶解于纯水中,加入乙醇,使乙醇含量达到40vol%,采用0.2M HCl和NaOH溶液调节pH值至4,搅拌,得到纤维基液;(4) Preparation of fiber-based liquid: dissolving the prepared turmeric water-soluble fiber in pure water, adding ethanol to make the ethanol content reach 40 vol%, adjusting the pH value to 4 with 0.2 M HCl and NaOH solution, stirring, and obtaining a fiber-based liquid;
其中,复合姜黄素液中的姜黄素与姜黄水溶性纤维的质量比为1:30;Among them, the mass ratio of curcumin to turmeric water-soluble fiber in the composite curcumin solution is 1:30;
(5)将复合姜黄素液滴加至搅拌状态中的纤维基液中,再滴加乙醇使乙醇含量达到40vol%,室温下于600rpm继续搅拌3h后,置于40℃水浴上旋转蒸发20min除去乙醇,用水定容至原体积,得到姜黄纤维制剂。(5) The composite curcumin liquid was added dropwise to the fiber base liquid in a stirring state, and ethanol was then added dropwise to make the ethanol content reach 40 vol%, and the mixture was stirred at 600 rpm for 3 h at room temperature, and then placed in a 40° C. water bath for 20 min to remove the ethanol by rotary evaporation, and the volume was adjusted to the original volume with water to obtain a turmeric fiber preparation.
实施例3Example 3
本实施例提供一种姜黄纤维制剂的制备方法,步骤包括:This embodiment provides a method for preparing a turmeric fiber preparation, the steps comprising:
(1)负载有姜黄素的蛋白-壳聚糖纳米复合物的制备:将220mg乳铁蛋白和210mgMw=200ku,脱乙酰度≥90%的壳聚糖分别溶解在两份30mL含有0.5%冰醋酸,6mol/L尿素的解离溶液中;用乙酸将最终pH值调至5.5,并在4℃下孵育12h以完全解离;将600mg姜黄素溶解在22mL乙醇中得到姜黄素乙醇溶液;将姜黄素乙醇溶液与乳铁蛋白溶液混合得到混合液一,将混溶液一和壳聚糖溶液等比例混合,使用分子质量为10ku的透析管在2.5℃下用去离子水透析2天以去除尿素;透析完成后得到混合液二,在8 000g条件下离心25min,收集上清液即得;(1) Preparation of protein-chitosan nanocomposites loaded with curcumin: 220 mg lactoferrin and 210 mg chitosan with Mw=200 ku and a deacetylation degree of ≥90% were dissolved in two 30 mL dissociation solutions containing 0.5% glacial acetic acid and 6 mol/L urea, respectively; the final pH value was adjusted to 5.5 with acetic acid, and the mixture was incubated at 4°C for 12 h for complete dissociation; 600 mg curcumin was dissolved in 22 mL ethanol to obtain a curcumin ethanol solution; the curcumin ethanol solution was mixed with the lactoferrin solution to obtain a mixed solution 1; the mixed solution 1 and the chitosan solution were mixed in equal proportions, and dialyzed with deionized water at 2.5°C for 2 days using a dialysis tube with a molecular weight of 10 ku to remove urea; after the dialysis was completed, a mixed solution 2 was obtained, which was centrifuged at 8,000 g for 25 min, and the supernatant was collected;
其中,10mg姜黄素(纯度>95%)包括姜黄素72.22wt%、去甲氧基姜黄素17.72wt%和双去甲氧基姜黄素5.26wt%。Among them, 10 mg of curcumin (purity>95%) includes 72.22 wt % of curcumin, 17.72 wt % of demethoxycurcumin and 5.26 wt % of bisdemethoxycurcumin.
(2)复合姜黄素液的制备:将负载有姜黄素的蛋白-壳聚糖纳米复合物溶解于无水乙醇中得到复合姜黄素液;复合姜黄素液的浓度为7mg/mL;(2) Preparation of composite curcumin solution: dissolving the protein-chitosan nanocomposite loaded with curcumin in anhydrous ethanol to obtain composite curcumin solution; the concentration of the composite curcumin solution is 7 mg/mL;
(3)姜黄水溶性纤维的制备:将姜黄纤维粉置于70℃水中,料液比为1:30,搅拌30分钟后,过滤;将所得滤液与4倍体积的乙醇混合并静置1天,于8000rpm离心15min,将沉淀冷冻干燥后,得到姜黄水溶性纤维;(3) Preparation of turmeric water-soluble fiber: turmeric fiber powder was placed in 70°C water with a solid-liquid ratio of 1:30, stirred for 30 minutes, and filtered; the obtained filtrate was mixed with 4 volumes of ethanol and allowed to stand for 1 day, centrifuged at 8000 rpm for 15 minutes, and the precipitate was freeze-dried to obtain turmeric water-soluble fiber;
(4)纤维基液的制备:将制备的姜黄水溶性纤维溶解于纯水中,加入乙醇,使乙醇含量达到40vol%,采用0.2M HCl和NaOH溶液调节pH值至4,搅拌,得到纤维基液;(4) Preparation of fiber-based liquid: dissolving the prepared turmeric water-soluble fiber in pure water, adding ethanol to make the ethanol content reach 40 vol%, adjusting the pH value to 4 with 0.2 M HCl and NaOH solution, stirring, and obtaining a fiber-based liquid;
其中,复合姜黄素液中的姜黄素与姜黄水溶性纤维的质量比为1:30;Among them, the mass ratio of curcumin to turmeric water-soluble fiber in the composite curcumin solution is 1:30;
(5)将复合姜黄素液滴加至搅拌状态中的纤维基液中,再滴加乙醇使乙醇含量达到40vol%,室温下于600rpm继续搅拌3h后,置于40℃水浴上旋转蒸发20min除去乙醇,用水定容至原体积,得到姜黄纤维制剂。(5) The composite curcumin liquid was added dropwise to the fiber base liquid in a stirring state, and ethanol was then added dropwise to make the ethanol content reach 40 vol%, and the mixture was stirred at 600 rpm for 3 h at room temperature, and then placed in a 40° C. water bath for 20 min to remove the ethanol by rotary evaporation, and the volume was adjusted to the original volume with water to obtain a turmeric fiber preparation.
实施例4Example 4
本实施例提供一种姜黄纤维制剂的制备方法,步骤包括:This embodiment provides a method for preparing a turmeric fiber preparation, the steps comprising:
(1)负载有姜黄素的蛋白-壳聚糖纳米复合物的制备:将200mg乳铁蛋白和210mgMw=200ku,脱乙酰度≥90%的壳聚糖分别溶解在两份25mL含有0.5%冰醋酸,6mol/L尿素的解离溶液中;用乙酸将最终pH值调至5.5,并在4℃下孵育12h以完全解离;将560mg姜黄素溶解在21mL乙醇中得到姜黄素乙醇溶液;将姜黄素乙醇溶液与乳铁蛋白溶液混合得到混合液一,将混溶液一和壳聚糖溶液等比例混合,使用分子质量为10ku的透析管在2.5℃下用去离子水透析2天以去除尿素;透析完成后得到混合液二,在8 000g条件下离心20min,收集上清液即得;(1) Preparation of protein-chitosan nanocomposites loaded with curcumin: 200 mg lactoferrin and 210 mg chitosan with Mw=200 ku and a deacetylation degree of ≥90% were dissolved in two 25 mL dissociation solutions containing 0.5% glacial acetic acid and 6 mol/L urea, respectively; the final pH value was adjusted to 5.5 with acetic acid, and the mixture was incubated at 4°C for 12 h for complete dissociation; 560 mg curcumin was dissolved in 21 mL ethanol to obtain a curcumin ethanol solution; the curcumin ethanol solution was mixed with the lactoferrin solution to obtain a mixed solution 1; the mixed solution 1 and the chitosan solution were mixed in equal proportions, and dialyzed with deionized water for 2 days at 2.5°C using a dialysis tube with a molecular weight of 10 ku to remove urea; after the dialysis was completed, a mixed solution 2 was obtained, which was centrifuged at 8,000 g for 20 min, and the supernatant was collected;
其中,10mg姜黄素(纯度>95%)包括姜黄素72.22wt%、去甲氧基姜黄素17.72wt%和双去甲氧基姜黄素5.26wt%。Among them, 10 mg of curcumin (purity>95%) includes 72.22 wt % of curcumin, 17.72 wt % of demethoxycurcumin and 5.26 wt % of bisdemethoxycurcumin.
(2)复合姜黄素液的制备:将负载有姜黄素的蛋白-壳聚糖纳米复合物溶解于无水乙醇中得到复合姜黄素液;复合姜黄素液的浓度为6mg/mL;(2) Preparation of composite curcumin solution: dissolving the protein-chitosan nanocomposite loaded with curcumin in anhydrous ethanol to obtain a composite curcumin solution; the concentration of the composite curcumin solution is 6 mg/mL;
(3)姜黄水溶性纤维的制备:将姜黄纤维粉置于70℃水中,料液比为1:30,搅拌30分钟后,过滤;将所得滤液与4倍体积的乙醇混合并静置1天,于8000rpm离心15min,将沉淀冷冻干燥后,得到姜黄水溶性纤维;(3) Preparation of turmeric water-soluble fiber: turmeric fiber powder was placed in 70°C water with a solid-liquid ratio of 1:30, stirred for 30 minutes, and filtered; the obtained filtrate was mixed with 4 volumes of ethanol and allowed to stand for 1 day, centrifuged at 8000 rpm for 15 minutes, and the precipitate was freeze-dried to obtain turmeric water-soluble fiber;
(4)纤维基液的制备:将制备的姜黄水溶性纤维溶解于纯水中,加入乙醇,使乙醇含量达到40vol%,采用0.2M HCl和NaOH溶液调节pH值至4,搅拌,得到纤维基液;(4) Preparation of fiber-based liquid: dissolving the prepared turmeric water-soluble fiber in pure water, adding ethanol to make the ethanol content reach 40 vol%, adjusting the pH value to 4 with 0.2 M HCl and NaOH solution, stirring, and obtaining a fiber-based liquid;
其中,复合姜黄素液中的姜黄素与姜黄水溶性纤维的质量比为1:30;Among them, the mass ratio of curcumin to turmeric water-soluble fiber in the composite curcumin solution is 1:30;
(5)将复合姜黄素液滴加至搅拌状态中的纤维基液中,再滴加乙醇使乙醇含量达到40vol%,室温下于600rpm继续搅拌3h后,置于40℃水浴上旋转蒸发20min除去乙醇,用水定容至原体积,得到姜黄纤维制剂。(5) The composite curcumin liquid was added dropwise to the fiber base liquid in a stirring state, and ethanol was then added dropwise to make the ethanol content reach 40 vol%, and the mixture was stirred at 600 rpm for 3 h at room temperature, and then placed in a 40° C. water bath for 20 min to remove the ethanol by rotary evaporation, and the volume was adjusted to the original volume with water to obtain a turmeric fiber preparation.
试验例2Test Example 2
依照前述试验例方法,对实施例2-3进行测试,并将实施例1数据作为对比,结果如下表3所示:According to the above test method, Examples 2-3 were tested, and the data of Example 1 was used as a comparison. The results are shown in Table 3 below:
表3DAI得分表(平均数±标准差,n=8)Table 3 DAI score table (mean ± standard deviation, n = 8)
最后应说明的是:以上实施例仅用以说明本发明而并非限制本发明所描述的技术方案;本领域的普通技术人员应当理解,仍然可以对本发明进行修改或等同替换;而一切不脱离本发明的精神和范围的技术方案及其改进,其均应涵盖在本发明的权利要求范围中。Finally, it should be noted that the above embodiments are only used to illustrate the present invention and are not intended to limit the technical solutions described in the present invention. Those skilled in the art should understand that the present invention can still be modified or replaced by equivalents. All technical solutions and improvements that do not depart from the spirit and scope of the present invention should be included in the scope of the claims of the present invention.
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Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20110190399A1 (en) * | 2008-07-31 | 2011-08-04 | Santosh Kumar Kar | Curcumin nanoparticles and methods of producing the same |
CN104273522A (en) * | 2013-07-03 | 2015-01-14 | 江南大学 | Curcumin nanor compound and preparation method thereof |
CN110089753A (en) * | 2019-04-16 | 2019-08-06 | 东北农业大学 | A kind of preparation method for the nanoparticle improving curcumin bioavailability |
CN110393295A (en) * | 2019-08-22 | 2019-11-01 | 哈尔滨工业大学 | A curcumin composite nanoparticle solution with antioxidant function and preparation method thereof |
KR20220086018A (en) * | 2020-12-16 | 2022-06-23 | 가톨릭대학교 산학협력단 | Nanocomposites for enhancing stability and bioavailability of curcumin using cellulose and preparation method thereof |
CN114712327A (en) * | 2022-05-05 | 2022-07-08 | 辽宁大学 | A kind of preparation method of curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles |
CN115607524A (en) * | 2022-10-17 | 2023-01-17 | 合肥工业大学 | A composite nanoparticle loaded with curcumin and its preparation method |
WO2023065528A1 (en) * | 2021-10-21 | 2023-04-27 | 琛蓝(美国)营养制品股份有限公司 | Curcumin composite composition and preparation method therefor |
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20110190399A1 (en) * | 2008-07-31 | 2011-08-04 | Santosh Kumar Kar | Curcumin nanoparticles and methods of producing the same |
CN104273522A (en) * | 2013-07-03 | 2015-01-14 | 江南大学 | Curcumin nanor compound and preparation method thereof |
CN110089753A (en) * | 2019-04-16 | 2019-08-06 | 东北农业大学 | A kind of preparation method for the nanoparticle improving curcumin bioavailability |
CN110393295A (en) * | 2019-08-22 | 2019-11-01 | 哈尔滨工业大学 | A curcumin composite nanoparticle solution with antioxidant function and preparation method thereof |
KR20220086018A (en) * | 2020-12-16 | 2022-06-23 | 가톨릭대학교 산학협력단 | Nanocomposites for enhancing stability and bioavailability of curcumin using cellulose and preparation method thereof |
WO2023065528A1 (en) * | 2021-10-21 | 2023-04-27 | 琛蓝(美国)营养制品股份有限公司 | Curcumin composite composition and preparation method therefor |
CN114712327A (en) * | 2022-05-05 | 2022-07-08 | 辽宁大学 | A kind of preparation method of curcumin-loaded rice bran protein-gum arabic-sodium caseinate nanoparticles |
CN115607524A (en) * | 2022-10-17 | 2023-01-17 | 合肥工业大学 | A composite nanoparticle loaded with curcumin and its preparation method |
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