WO2020224554A1 - Folate-metal ion composition, preparation method therefor and use thereof in sun protection - Google Patents

Folate-metal ion composition, preparation method therefor and use thereof in sun protection Download PDF

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WO2020224554A1
WO2020224554A1 PCT/CN2020/088465 CN2020088465W WO2020224554A1 WO 2020224554 A1 WO2020224554 A1 WO 2020224554A1 CN 2020088465 W CN2020088465 W CN 2020088465W WO 2020224554 A1 WO2020224554 A1 WO 2020224554A1
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folic acid
hydrogel
metal ion
zinc
aqueous solution
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阎云
刘卡尔顿
臧士豪
黄建滨
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北京大学
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
    • A61K8/04—Dispersions; Emulsions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/19—Cosmetics or similar toiletry preparations characterised by the composition containing inorganic ingredients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/19—Cosmetics or similar toiletry preparations characterised by the composition containing inorganic ingredients
    • A61K8/27—Zinc; Compounds thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/67—Vitamins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G83/00—Macromolecular compounds not provided for in groups C08G2/00 - C08G81/00
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
    • C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
    • C08J3/075—Macromolecular gels
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
    • C08J3/00—Processes of treating or compounding macromolecular substances
    • C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor

Abstract

A folate-metal ion composition, a folate-metal ion hydrogel, a sun-screening agent containing the hydrogel and use thereof in ultraviolet absorption. After a folate aqueous solution and a metal ion aqueous solution have been mixed, folate and the metal ions are cross-linked to form a solution, gel or suspension containing a reticular hydrogel, and the metal ions are coordinated with folate molecules. The folate-metal ion hydrogel has good biocompatibility, has significant advantages in the field of biological applications, and also provides a new nutritional supplement form. The sun protection effect of the folate-metal ion hydrogel and folate are almost constant. The folate-metal ion hydrogel has good safety, high reliability and stable chemical properties, and is refreshing after being prepared into a sun-screening agent, having good user experience. The preparation method for the folate-metal ion hydrogel is simple and convenient, controllable and adjustable, and can satisfy different production requirements and consumer demands.

Description

叶酸-金属离子组合物及其制备方法和在防晒中的用途Folic acid-metal ion composition and its preparation method and use in sunscreen 技术领域Technical field
本发明涉及水凝胶技术领域，特别涉及金属离子和叶酸配位形成的水凝胶及其在防晒中的用途。The invention relates to the technical field of hydrogels, in particular to hydrogels formed by the coordination of metal ions and folic acid and their use in sun protection.
背景技术Background technique
太阳光约由5％紫外线(200-400nm)、43％可见光(400-700nm)、52％近红外(700-2500nm)组成，其中紫外线可以分为短波紫外线(200-275nm)、中波紫外线(275-320nm)、长波紫外线(320-400nm)。短波紫外线(UVC)几乎都会被大气层滤掉；中波紫外线(UVB)会穿透角皮层，引起大多数皮肤伤害；长波紫外线(UVA)可以穿透真皮，使真皮产生黑色素，造成皮肤晒黑老化或晒红晒伤。Sunlight is approximately composed of 5% ultraviolet (200-400nm), 43% visible light (400-700nm), 52% near infrared (700-2500nm), among which ultraviolet rays can be divided into short-wave ultraviolet (200-275nm) and medium-wave ultraviolet ( 275-320nm), long-wave ultraviolet (320-400nm). Short-wave ultraviolet (UVC) is almost always filtered out by the atmosphere; medium-wave ultraviolet (UVB) penetrates the stratum corneum and causes most skin damage; long-wave ultraviolet (UVA) can penetrate the dermis, causing the dermis to produce melanin, causing skin tanning and aging Or sunburn.
传统的防晒原理主要分为两类：Traditional sun protection principles are mainly divided into two categories:
1.物理防晒：主要成分是氧化锌和二氧化钛，其可以停留在皮肤表面并反射和散射紫外线，通过使到达皮肤的紫外线量减少来达到防晒目的。二氧化钛可完全阻隔UVB，但只能隔绝波长较短的UVA。氧化锌几乎可以阻隔所有波长的UVA和UVB，且安全性较高，但是涂抹在皮肤上会发白，且较为粘腻。1. Physical sunscreen: The main ingredients are zinc oxide and titanium dioxide, which can stay on the surface of the skin and reflect and scatter ultraviolet rays to achieve sun protection by reducing the amount of ultraviolet rays reaching the skin. Titanium dioxide can completely block UVB, but it can only block UVA with a shorter wavelength. Zinc oxide can block almost all wavelengths of UVA and UVB, and it is safer, but it will be white and sticky when applied to the skin.
2.化学防晒。化学吸光剂通过其他成膜性物质在肌肤表面形成一道紫外线吞噬屏障，吸收中和紫外线，防止其射入皮肤造成伤害。2. Chemical sunscreen. Chemical light absorbers form an ultraviolet phagocytic barrier on the skin surface through other film-forming substances, absorb and neutralize ultraviolet rays, and prevent it from entering the skin and causing damage.
当前，防晒领域的发展方向主要分为三个方面：At present, the development direction of the sunscreen field is mainly divided into three aspects:
1.有效性：防晒系数(SPF)是指防晒品对阳光中紫外线的防御能力的检测指数，表明防晒用品所能发挥的防晒效能的高低。它根据皮肤的最低红斑剂量来确定,即是皮肤出现红斑的最短日晒时间。SPF防晒系数的数值意味着防晒品可以在相同的紫外线强度下延长多少倍的抵抗紫外线时间。使用防晒用品后，皮肤的最低红斑剂量会增长，防晒系数SPF为：SPF＝最低红斑剂量(使用后)/最低红斑剂量(使用前)，当前研发方向自然希望发展更高SPF等级的防晒产品。1. Effectiveness: The sun protection factor (SPF) refers to the detection index of the sunscreen's defense against ultraviolet rays in the sun, indicating the level of sunscreen effectiveness that the sunscreen can exert. It is determined according to the minimum erythema dose of the skin, that is, the shortest sun exposure time for erythema on the skin. The value of the SPF sun protection factor means how many times the sunscreen can prolong the UV resistance time under the same UV intensity. After using sunscreen products, the minimum erythema dose of the skin will increase. The sun protection factor SPF is: SPF=minimum erythema dose (after use)/minimum erythema dose (before use). The current research and development direction naturally hopes to develop sunscreen products with higher SPF levels.
2.安全性：一些化学防晒剂如阿伏苯宗(Avobenzone、BMDM)、甲氧基肉桂酸辛酯(OMC)等，日光照射分解后产生的自由基或者分子碎片会破坏细胞和组织。另外，一些防晒成份会被皮肤吸收，在皮肤内吸收紫外线，虽然过后可以被人体代谢清除，但也会有安全健康的忧虑，甚至会令皮肤产生依赖性。研发人员希望得到更安全、稳定的化学防晒剂。2. Safety: some chemical sunscreen agents such as Avobenzone (BMDM), octyl methoxycinnamate (OMC), etc., free radicals or molecular fragments generated after the decomposition of sunlight can damage cells and tissues. In addition, some sunscreen ingredients will be absorbed by the skin and absorb ultraviolet rays in the skin. Although it can be eliminated by the body's metabolism, there will be concerns about safety and health, and even skin dependence. Researchers hope to obtain safer and more stable chemical sunscreens.
化学防晒剂的安全问题在研发中尤为重要。FDA在2019年组织志愿者进行测试(JAMA.2019；321(21):2082-2091.doi:10.1001/jama.2019.5586)，以四次每天，连续涂四天的频率涂防晒霜，检测血浆中防晒剂的组分。结果表明，四种常见的商用化学防晒成分，阿伏苯宗、二苯甲酮-3(BP-3)、水杨酸辛酯(Octyl salicylate)、甲氧基肉桂酸辛酯，在血浆中的浓度均超过了FDA规定的血液中该成分的最高浓度。The safety of chemical sunscreens is particularly important in research and development. In 2019, the FDA organized volunteers for testing (JAMA.2019; 321(21):2082-2091.doi:10.1001/jama.2019.5586), applying sunscreen four times a day for four consecutive days to detect plasma Components of sunscreens. The results showed that the plasma concentrations of four common commercial chemical sunscreen ingredients, Avobenzone, Benzophenone-3 (BP-3), Octyl Salicylate, and Octyl Methoxycinnamate Both exceed the maximum concentration of this component in the blood specified by the FDA.
3.消费者接受程度：传统的防晒剂为霜或乳膏等，在炎热的夏天，过于油腻的产品会增加用户的闷热感。消费者会对新型的喷雾、凝胶等剂型的防晒剂产生兴趣。3. Consumer acceptance: traditional sunscreens are creams or creams, etc. In the hot summer, too greasy products will increase the user's sultry feeling. Consumers will be interested in new types of sunscreens such as sprays and gels.
水凝胶是一类在水中形成的软物质材料，由于其具有类似生物组织的柔软和湿润的性质，近年来得到了越来越多的研究者的关注。Hydrogel is a kind of soft material formed in water. Due to its soft and moist properties similar to biological tissues, it has attracted more and more researchers' attention in recent years.
水凝胶可以作为生物材料进行利用，例如利用水凝胶作为细胞培育的基质、胶制备组织器官植入体等。在水凝胶的生物利用中，需要考虑的指标除了传统的机械性质(强度、硬度、模量等)外，生物相容性更是其中极为重要的考量指标。例如，作为组织培养基质的水凝胶必须不含有影响细胞孵育的有害组分；作为器官植入体的水凝胶除了需要本身对生物体无害外，还需要考虑其降解产物是否对生物体不利。因而完全生物相容性的水凝胶成为水凝胶生物应用中的重要问题。Hydrogels can be used as biomaterials, for example, using hydrogels as a matrix for cell cultivation, glues to prepare tissue organ implants, and the like. In the bioavailability of hydrogels, in addition to the traditional mechanical properties (strength, hardness, modulus, etc.), biocompatibility is an extremely important consideration. For example, the hydrogel used as a tissue culture substrate must not contain harmful components that affect cell incubation; the hydrogel used as an organ implant not only needs to be harmless to the organism itself, but also needs to consider whether its degradation products are harmful to the organism. unfavorable. Therefore, a completely biocompatible hydrogel has become an important issue in the biological application of hydrogels.
在生物相容性的水凝胶研究中，很自然的一个想法就是利用生物体内已有的自然物质构建水凝胶，这样“取之自然，用之自然”得到的水凝胶也就自然而然地具备生物相容性。In the research of biocompatibility hydrogels, a natural idea is to construct hydrogels using natural substances already in the organism. In this way, the hydrogels obtained by "take nature and use nature" will naturally With biocompatibility.
然而，目前大多数的水凝胶来源于合成的高分子(Pal,K.,Banthia,A.K.,&Majumdar,D.K.(2009).Polymeric hydrogels:characterization and biomedical applications.Designed monomers and polymers,12(3),197-220.)，少部分水凝胶来源于小分子量分子，其中利用的分子也是经过特殊的设计和合成得到(De Loos,M.,Feringa,B.L.,& van Esch,J.H.(2005).Design and application of self‐assembled low molecular weight hydrogels.European Journal of Organic Chemistry,2005(17),3615-3631.)。而利用生物已有的自然分子构建水凝胶的体系少之又少。倘若我们能够利用生物分子简易地构建水凝胶，那么这种生物相容性极强的水凝胶将在生物应用中具备广阔的应用前景。However, most of the current hydrogels are derived from synthetic polymers (Pal, K., Banthia, AK, & Majumdar, DK (2009). Polymeric hydrogels: characterization and biomedical applications. Designed monomers and polymers, 12(3), 197-220.), a small number of hydrogels are derived from small molecular weight molecules, and the molecules used are also obtained through special design and synthesis (De Loos, M., Feringa, BL, & van Esch, JH (2005). Design and application of self-assembled low molecular weight hydrogels. European Journal of Organic Chemistry, 2005(17), 3615-3631.). However, there are very few systems that use the existing natural molecules of organisms to construct hydrogels. If we can easily construct hydrogels using biomolecules, then this highly biocompatible hydrogel will have broad application prospects in biological applications.
因此，本发明提供了一种基于自然已有分子水溶性维生素B9叶酸构建水凝胶的方法，以及该水凝胶在防晒中的用途。Therefore, the present invention provides a method for constructing a hydrogel based on the natural existing molecule water-soluble vitamin B9 folic acid, and the use of the hydrogel in sun protection.
发明内容Summary of the invention
为了解决上述问题，本发明人进行了锐意研究，结果发现：基于自然已有分子叶酸构建的水凝胶具有良好的生物相容性，通过水溶性的叶酸溶液和金属离子溶液的均匀混合，使叶酸和金属离子相互配位交联，形成水凝胶。叶酸和金属离子制备为水凝胶后可以显著改善叶酸溶解性差的问题，使其以分子状态存在于水凝胶中，使水凝胶的防晒效果几乎与单独的叶酸溶液保持恒定。制备得到的叶酸-金属离子水 凝胶可以很容易调控性状，在一定叶酸-金属离子比例范围内可以得到流体性状不同的水凝胶。该水凝胶具有凝胶的粘稠性、涂展性和均匀性，安全性好可靠性高，化学性质稳定，制备为防晒剂后非常清爽，使用体验良好，从而完成了本发明。本发明的目的在于提供以下方面：In order to solve the above-mentioned problems, the inventors conducted intensive research and found that: the hydrogel constructed based on the naturally existing molecular folic acid has good biocompatibility. The water-soluble folic acid solution and the metal ion solution are uniformly mixed to make Folic acid and metal ions coordinate and cross-link each other to form a hydrogel. After folic acid and metal ions are prepared as a hydrogel, the problem of poor solubility of folic acid can be significantly improved, so that it exists in the hydrogel in a molecular state, so that the sunscreen effect of the hydrogel is almost constant with that of the folic acid solution alone. The prepared folic acid-metal ion hydrogel can easily control the properties, and within a certain range of folic acid-metal ion ratio, hydrogels with different fluid properties can be obtained. The hydrogel has the viscosity, spreadability and uniformity of a gel, has good safety, high reliability, stable chemical properties, is very refreshing after being prepared as a sunscreen, and has a good use experience, thereby completing the present invention. The purpose of the present invention is to provide the following aspects:
第一方面，本发明提供一种叶酸-金属离子水凝胶，该水凝胶具有网状结构，包括配位连接的金属离子和叶酸分子；其中，叶酸的喋呤头基通过氢键形成四联体，四联体之间π-π堆积形成纤维，金属离子和叶酸的羧酸配位，使纤维交联并相互缠绕形成网状结构。In the first aspect, the present invention provides a folic acid-metal ion hydrogel, which has a network structure, including coordinated connection of metal ions and folic acid molecules; wherein the pterin head group of folic acid forms four through hydrogen bonding. Conjoined, π-π stacked between tetrads form fibers, and the metal ions are coordinated with the carboxylic acid of folic acid, so that the fibers are cross-linked and intertwined to form a network structure.
本发明还提供了一种叶酸-金属离子水凝胶，该水凝胶由包括以下步骤制备：The present invention also provides a folic acid-metal ion hydrogel, which is prepared by including the following steps:
(1)将叶酸溶解于水或水性缓冲溶液中，配制叶酸水溶液；(1) Dissolve folic acid in water or an aqueous buffer solution to prepare an aqueous folic acid solution;
(2)将金属盐溶解于水或水性缓冲溶液中，配制金属离子水溶液；(2) Dissolve the metal salt in water or an aqueous buffer solution to prepare an aqueous solution of metal ions;
(3)将叶酸水溶液和金属离子水溶液室温下均匀混合，静置，即得叶酸-金属离子水凝胶；(3) The folic acid aqueous solution and the metal ion aqueous solution are uniformly mixed at room temperature and left to stand to obtain folic acid-metal ion hydrogel;
所述叶酸-金属离子水凝胶为溶液、凝胶或混悬液状态。The folic acid-metal ion hydrogel is in the state of solution, gel or suspension.
第二方面，本发明提供了一种防晒剂，其含有上述叶酸-金属离子水凝胶，所述防晒剂中还包括氧化锌；In a second aspect, the present invention provides a sunscreen, which contains the above-mentioned folic acid-metal ion hydrogel, and the sunscreen also includes zinc oxide;
所述氧化锌在防晒剂中的质量分数为0.1％-10％，优选为0.5％-5％；所述防晒剂为膏剂、霜剂或凝胶剂。The mass fraction of the zinc oxide in the sunscreen is 0.1%-10%, preferably 0.5%-5%; the sunscreen is an ointment, cream or gel.
第三方面，本发明提供了第一方面叶酸-金属离子水凝胶的制备方法，包括以下步骤：In the third aspect, the present invention provides the preparation method of folic acid-metal ion hydrogel in the first aspect, which includes the following steps:
(1)配制叶酸水溶液；(1) Prepare an aqueous solution of folic acid;
(2)配制金属离子水溶液；(2) Preparation of metal ion aqueous solution;
(3)将叶酸水溶液和金属离子水溶液室温下均匀混合，静置，即得水凝胶。(3) The folic acid aqueous solution and the metal ion aqueous solution are uniformly mixed at room temperature and left to stand to obtain a hydrogel.
其中，在步骤(1)中，将叶酸溶解于水性溶液中配制叶酸水溶液；所述水性溶液包括水或水性缓冲溶液；和/或，Wherein, in step (1), folic acid is dissolved in an aqueous solution to prepare an aqueous folic acid solution; the aqueous solution includes water or an aqueous buffer solution; and/or,
在步骤(2)中，所述金属离子选自一价或二价态时化学性质稳定的金属离子；优选地，所述金属离子选自钠、钾、锂、铷、铯、锌(Zn 2+)，铜(Cu 2+)，镍(Ni 2+)，钴(Co 2+)，锰(Mn 2+)，铬(Cr 2+)，钼(Mo 2+)，银(Ag +)，镉(Cd 2+)，镁(Mg 2+)，钙(Ca 2+)，铅(Pb 2+)、钡、金属中的一种或多种；和/或， In step (2), the metal ion is selected from metal ions that are chemically stable in a monovalent or divalent state; preferably, the metal ion is selected from sodium, potassium, lithium, rubidium, cesium, zinc (Zn 2 + ), copper (Cu 2+ ), nickel (Ni 2+ ), cobalt (Co 2+ ), manganese (Mn 2+ ), chromium (Cr 2+ ), molybdenum (Mo 2+ ), silver (Ag + ) , Cadmium (Cd 2+ ), magnesium (Mg 2+ ), calcium (Ca 2+ ), lead (Pb 2+ ), barium, one or more of metals; and/or,
在步骤(3)中，混合后，所述叶酸在混合体系中的最终浓度为0.5mM-200mM，优选为5mM-100mM；In step (3), after mixing, the final concentration of the folic acid in the mixing system is 0.5mM-200mM, preferably 5mM-100mM;
所述金属离子在混合体系中的最终浓度为0.5mM-500mM，优选为5mM-400mM。The final concentration of the metal ion in the mixed system is 0.5mM-500mM, preferably 5mM-400mM.
第四方面，本发明提供了第一方面叶酸-金属离子水凝胶或第三方面所述方法制备的水凝胶在生物材料中的用途；优选在细胞三维培养中的用途或生物三维打印中的用途。In the fourth aspect, the present invention provides the use of the folic acid-metal ion hydrogel in the first aspect or the hydrogel prepared by the method in the third aspect in biomaterials; preferably in three-dimensional cell culture or three-dimensional bioprinting the use of.
第五方面，本发明提供了第一方面叶酸-金属离子水凝胶和第二方面防晒剂在预防或防止紫外吸收中的用途；优选用作防晒剂的用途；和/或，In the fifth aspect, the present invention provides the use of the folic acid-metal ion hydrogel in the first aspect and the use of the sunscreen in the second aspect in preventing or preventing ultraviolet absorption; preferably as a sunscreen; and/or,
叶酸-金属离子组合物在预防或防止紫外吸收中的用途，其特征在于，所述叶酸-金属离子组合物包括叶酸，和锌、银、镁和钙离子中的一种或多种；The use of a folic acid-metal ion composition for preventing or preventing ultraviolet absorption, characterized in that the folic acid-metal ion composition includes folic acid, and one or more of zinc, silver, magnesium and calcium ions;
优选所述叶酸-金属离子组合物为叶酸-金属离子水凝胶。Preferably, the folic acid-metal ion composition is folic acid-metal ion hydrogel.
根据本发明提供的叶酸-金属离子组合物及其制备方法和在防晒中的用途，具有以下有益效果：According to the folic acid-metal ion composition provided by the present invention, its preparation method and use in sunscreen, it has the following beneficial effects:
(1)所述叶酸-金属离子水凝胶制备方法简便、可控和易调，并且易于扩大化生产，水凝胶的性质可以很容易地被调控，从而获得不同需求不同性质的水凝胶。(1) The preparation method of the folic acid-metal ion hydrogel is simple, controllable and easy to adjust, and is easy to scale up production. The properties of the hydrogel can be easily adjusted to obtain hydrogels with different requirements and properties .
(2)所述叶酸-金属离子水凝胶具有良好的生物相容性，在生物应用领域具有显著优势，可以作为生物材料在细胞三维培养和生物三维打印中进行应用。(2) The folic acid-metal ion hydrogel has good biocompatibility, has significant advantages in the field of biological applications, and can be used as a biological material in three-dimensional cell culture and biological three-dimensional printing.
(3)所述叶酸金属离子水凝胶同时可以提供补充叶酸和微量元素的营养补充功效。(3) The folic acid metal ion hydrogel can also provide the nutritional supplement effect of supplementing folic acid and trace elements.
(4)将叶酸加入金属离子制备成水凝胶后，其防晒效果几乎保持恒定，并且叶酸-金属离子水凝胶可以很容易调控其性状，在一定叶酸-金属离子比例范围内可以得到水凝胶。该水凝胶性质可控，可以满足不同的生产需求和消费者需要。(4) After folic acid is added to metal ions to prepare a hydrogel, its sunscreen effect remains almost constant, and the folic acid-metal ion hydrogel can easily control its properties, and it can get hydrogel within a certain range of folic acid-metal ion ratio. glue. The hydrogel has controllable properties and can meet different production requirements and consumer needs.
(5)将叶酸制备为叶酸-金属离子水凝胶后具有粘稠性、涂展性和均匀性，非常有利于防晒剂的制备和成型。(5) Folic acid is prepared into folic acid-metal ion hydrogel with viscosity, spreadability and uniformity, which is very beneficial to the preparation and molding of sunscreen.
(6)叶酸-金属离子水凝胶外观和性状上类似市售的芦荟胶，相比于传统的油腻性防晒霜，水凝胶的消费者体验明显提高，可以提高消费者的接受程度。(6) The appearance and properties of the folic acid-metal ion hydrogel are similar to those of commercially available aloe vera gel. Compared with traditional greasy sunscreens, the consumer experience of the hydrogel is significantly improved, which can increase consumer acceptance.
(7)叶酸-金属离子水凝胶可以和其他化学紫外吸收剂、物理防晒剂复配，提高防晒效果。(7) Folic acid-metal ion hydrogel can be compounded with other chemical UV absorbers and physical sunscreens to improve the sun protection effect.
(8)所述叶酸-金属离子水凝胶安全性好可靠性高，化学性质稳定，制备为防晒剂后非常清爽，使用体验良好。(8) The folic acid-metal ion hydrogel has good safety, high reliability, stable chemical properties, is very refreshing after being prepared as a sunscreen, and has a good user experience.
附图说明Description of the drawings
图1示出实施例1-3制得的水凝胶的宏观状态图；图2示出实验例1中三维打印的示意图；图3示出叶酸分子和金属离子形成四联体的示意图；图4示出叶酸分子和金属离子形成四联体后堆积形成纤维的示意图；图5示出实验例3中不同叶酸和锌离子比例水凝胶的流变表征振幅扫描图；图6示出实验例3中不同叶酸和锌离子比例水凝胶的流变表征频率扫描图；图7示出实验例4中叶酸-锌水凝胶(a)和叶酸-铜水凝胶(b)的透射 电镜图；图8示出实验例4中叶酸-锌水凝胶(a)和叶酸-铜水凝胶(b)的扫描电镜图；图9示出实验例5中叶酸-锌水凝胶和叶酸-铜水凝胶的XRD图；图10示出实验例6中叶酸-锌水凝胶的圆二色谱图；图11示出实验例2中叶酸-锌水凝胶加入hRPE细胞培养液中的细胞存活率；图12示出实验例2中叶酸-锌水凝胶加入HeLa细胞培养液中的细胞存活率；图13示出实验例2中叶酸-锌水凝胶三维培养hRPE细胞的细胞存活率；图14示出实验例2中叶酸-锌水凝胶三维培养HeLa细胞的细胞存活率；图15示出实验例7中化学吸收剂的紫外吸收图谱；图16示出实验例8中不同叶酸-锌离子比例的叶酸-锌水凝胶的紫外吸收图谱；图17示出实验例9中不同用量的阿伏苯宗、二苯甲酮-3、水杨酸异辛酯、甲氧基肉桂酸辛酯和实施例4制备的水凝胶的防晒效果比较图；图18示出实验例10中不同用量的氧化锌及氧化锌与实施例4制备的水凝胶复配的防晒效果比较图；图19示出实验例11中商用防晒剂和氧化锌与实施例1制备的水凝胶复配的防晒效果比较图；图20示出叶酸-锌离子的相图，包括溶液、凝胶和悬浊液。Figure 1 shows the macroscopic state diagram of the hydrogels prepared in Examples 1-3; Figure 2 shows the schematic diagram of the three-dimensional printing in Experimental Example 1; Figure 3 shows the schematic diagram of the formation of folic acid molecules and metal ions into quadruplets; 4 shows a schematic diagram of folic acid molecules and metal ions forming a quadruple and then accumulates to form fibers; Figure 5 shows the rheological characterization amplitude scan of the hydrogel with different folic acid and zinc ion ratios in Experimental Example 3; Figure 6 shows the experimental example Frequency scanning diagram of rheological characterization of hydrogels with different folic acid and zinc ion ratios in 3; Figure 7 shows the transmission electron micrographs of folic acid-zinc hydrogel (a) and folic acid-copper hydrogel (b) in Experimental Example 4 Figure 8 shows the scanning electron micrographs of folic acid-zinc hydrogel (a) and folic acid-copper hydrogel (b) in Experimental Example 4; Figure 9 shows the folic acid-zinc hydrogel and folic acid in Experimental Example 5 XRD pattern of the copper hydrogel; Figure 10 shows the circular dichroism of the folic acid-zinc hydrogel in Experimental Example 6; Figure 11 shows the cells of the folic acid-zinc hydrogel in Experimental Example 2 added to the hRPE cell culture solution Survival rate; Figure 12 shows the cell survival rate of folic acid-zinc hydrogel added to HeLa cell culture medium in Experimental Example 2; Figure 13 shows the cell survival rate of folic acid-zinc hydrogel three-dimensional cultured hRPE cells in Experimental Example 2 Figure 14 shows the cell viability of folic acid-zinc hydrogel three-dimensional culture HeLa cells in Experimental Example 2; Figure 15 shows the UV absorption spectrum of the chemical absorbent in Experimental Example 7; Figure 16 shows the different folic acid in Experimental Example 8. -Ultraviolet absorption spectrum of folic acid-zinc hydrogel with zinc ion ratio; Figure 17 shows different amounts of avobenzone, benzophenone-3, isooctyl salicylate, and methoxycinnamon in experimental example 9 The sunscreen effect comparison diagram of octyl acid and the hydrogel prepared in Example 4; FIG. 18 shows the sunscreen effect comparison diagram of different amounts of zinc oxide and zinc oxide in Experimental Example 10 and the hydrogel prepared in Example 4; Figure 19 shows the comparison of the sunscreen effect of the commercial sunscreen and zinc oxide in Experimental Example 11 and the hydrogel prepared in Example 1; Figure 20 shows the phase diagram of folic acid-zinc ions, including solution, gel and suspension Turbid liquid.
具体实施方式Detailed ways
下面通过对本发明进行详细说明，本发明的特点和优点将随着这些示例性说明而变得更为清楚、明确。在这里专用的词“示例性”意为“用作例子、实施例或说明性”。这里作为“示例性”所说明的任何实施例不必解释为优于或好于其它实施例。以下详述本发明。Through the detailed description of the present invention below, the characteristics and advantages of the present invention will become clearer and clearer with these exemplary descriptions. The dedicated word "exemplary" here means "serving as an example, embodiment, or illustration." Any embodiment described herein as "exemplary" need not be construed as being superior or better than other embodiments. The present invention will be described in detail below.
研究发现，叶酸在紫外线光区范围内可以吸收紫外线。在UV-B区域，叶酸具有和甲氧基肉桂酸辛酯(OMC)、二苯甲酮-3(BP-3)相比拟的吸光系数；在UV-A区域，叶酸也具有一定程度的吸光。因而叶酸可以作为UV-A、UV-B双区域的紫外吸光剂。Studies have found that folic acid can absorb ultraviolet light within the range of ultraviolet light. In the UV-B region, folic acid has an absorption coefficient comparable to that of octyl methoxycinnamate (OMC) and benzophenone-3 (BP-3); in the UV-A region, folic acid also has a certain degree of absorption. Therefore, folic acid can be used as a UV-A and UV-B dual-region ultraviolet absorber.
基于此，本发明提供了一种叶酸在紫外吸收剂中的用途，优选在UV-A和UV-B区域紫外吸收剂中的用途，更优选在防晒中的用途。Based on this, the present invention provides a use of folic acid in ultraviolet absorbers, preferably in UV-A and UV-B region ultraviolet absorbers, and more preferably in sunscreen.
叶酸为水溶性维生素B9，其作为预防胎儿神经管畸形的药物或添加剂已广泛应用于孕妇群体中，安全性非常高。叶酸具有良好的生物相容性，其光照产物不具有生物毒性。将叶酸应用于皮肤表面被吸收后，不会产生生物毒性，吸收的叶酸还可以参与人体内正常的叶酸循环代谢，甚至一定程度起到补充叶酸的功能。使用叶酸作为化学吸收活性成分的紫外吸收剂，相较现有技术中的化学防晒剂，安全性更高，对皮肤健康影响更小。Folic acid is a water-soluble vitamin B9, which has been widely used in pregnant women as a medicine or additive to prevent fetal neural tube malformations, and its safety is very high. Folic acid has good biocompatibility, and its light products are not biotoxic. After folic acid is applied to the skin surface and absorbed, it will not produce biological toxicity. The absorbed folic acid can also participate in the normal folic acid cycle metabolism in the human body, and even play a role in supplementing folic acid to a certain extent. The use of folic acid as an ultraviolet absorber for chemically absorbing active ingredients has higher safety and less impact on skin health than the chemical sunscreen in the prior art.
为了进一步提高叶酸紫外吸收或防紫外线的效果，还可以与ZnO和/或TiO 2等物理防晒的无机组分进行复配，进一步提高防晒效果。 In order to further improve the ultraviolet absorption or anti-ultraviolet effect of folic acid, it can also be compounded with ZnO and/or TiO 2 and other physical sunscreen inorganic components to further improve the sunscreen effect.
考虑到叶酸在水溶性和油溶性溶剂中的溶解性均较差，而且常规的霜或乳膏容易造成皮肤油腻感，本发明提供了一种较为清爽的叶酸组合物，尤其是叶酸-金属离子组合物，以用于紫外吸收，特别用于防晒。Considering that folic acid has poor solubility in water-soluble and oil-soluble solvents, and conventional creams or creams are likely to cause skin greasy feeling, the present invention provides a relatively refreshing folic acid composition, especially folic acid-metal ion The composition is used for ultraviolet absorption, especially for sun protection.
优选所述叶酸-金属离子组合物为叶酸-金属离子水凝胶。本发明中，所述叶酸-金属离子水凝胶是叶酸分子和金属离子充分交联配位形成的具有网状结构水凝胶分子的配合物。根据网状结构水凝胶分子在水中分散性的不同，所述叶酸-金属离子水凝胶具有溶液、凝胶或悬浮液等流体状态。Preferably, the folic acid-metal ion composition is folic acid-metal ion hydrogel. In the present invention, the folate-metal ion hydrogel is a complex of hydrogel molecules with a network structure formed by fully cross-linking and coordination of folate molecules and metal ions. According to the different dispersibility of the network structure hydrogel molecules in water, the folic acid-metal ion hydrogel has a fluid state such as a solution, a gel, or a suspension.
所述叶酸-金属离子水凝胶的制备包括以下步骤：The preparation of the folic acid-metal ion hydrogel includes the following steps:
(1)配制叶酸水溶液；(1) Prepare an aqueous solution of folic acid;
(2)配制金属离子水溶液；(2) Preparation of metal ion aqueous solution;
(3)将叶酸水溶液和金属离子水溶液室温下均匀混合，静置，即得叶酸-金属离子水凝胶。(3) The folic acid aqueous solution and the metal ion aqueous solution are uniformly mixed at room temperature and left to stand to obtain the folic acid-metal ion hydrogel.
其中，在步骤(1)中，优选将叶酸溶解于水性溶液中配制叶酸水溶液。所述水性溶液包括水或水性缓冲溶液。Among them, in step (1), it is preferable to dissolve folic acid in an aqueous solution to prepare an aqueous folic acid solution. The aqueous solution includes water or an aqueous buffer solution.
由于水中可能含有非所需的阳离子和阴离子，影响本发明中叶酸和设定金属离子形成配合物。优选在本发明中所用到的水都为不含有离子的纯化水，更优选为超纯水。Since water may contain undesired cations and anions, it affects the formation of complexes between folic acid and set metal ions in the present invention. Preferably, all the water used in the present invention is purified water that does not contain ions, and more preferably ultrapure water.
使用纯水制备叶酸水溶液时，为了提高叶酸的溶解性和溶解速度，滴加碱性试剂使水溶液的pH呈碱性，优选为7.0～9.5。When preparing an aqueous folic acid solution using pure water, in order to increase the solubility and dissolution rate of folic acid, an alkaline agent is added dropwise to make the pH of the aqueous solution alkaline, preferably 7.0 to 9.5.
所述碱性试剂选自氨水、碳酸钠、碳酸氢钠、碳酸钾、氢氧化钠、氢氧化钾、氢氧化锂、氢氧化铷、氢氧化铯中的一种或多种；优选为氢氧化钠、氢氧化钾、氢氧化锂、氢氧化铷和氢氧化铯中的一种或多种；更优选为氢氧化钠或氢氧化钾。The alkaline reagent is selected from one or more of ammonia, sodium carbonate, sodium bicarbonate, potassium carbonate, sodium hydroxide, potassium hydroxide, lithium hydroxide, rubidium hydroxide, and cesium hydroxide; preferably, it is hydroxide One or more of sodium, potassium hydroxide, lithium hydroxide, rubidium hydroxide, and cesium hydroxide; more preferably sodium hydroxide or potassium hydroxide.
使用氢氧化物和叶酸中的羧基反应时，不会增加叶酸水溶液中无机阴离子的种类和含量，尽量减少叶酸和金属离子配位的干扰。When the hydroxide is used to react with the carboxyl group in folic acid, the type and content of inorganic anions in the aqueous folic acid solution will not increase, and the interference of the coordination of folic acid and metal ions will be minimized.
另外，研究发现，使用氢氧化钠、氢氧化钾、氢氧化锂、氢氧化铷和氢氧化铯中的一种或多种调节溶液pH值可以向叶酸水溶液中添加钠、钾、锂、铷和/或铯金属离子。钠、钾、锂、铷和/或铯金属离子在水凝胶的形成过程中对叶酸和金属离子形成的四连体可能起到一定程度稳定的作用。In addition, studies have found that using one or more of sodium hydroxide, potassium hydroxide, lithium hydroxide, rubidium hydroxide and cesium hydroxide to adjust the pH of the solution can add sodium, potassium, lithium, rubidium and / Or cesium metal ion. Sodium, potassium, lithium, rubidium and/or cesium metal ions may have a certain degree of stability in the formation of folic acid and metal ions during the formation of the hydrogel.
所述水性缓冲溶液选自乙酸-乙酸钠、硼酸-硼砂、HEPES(N-(2-羟乙基)哌嗪-N'-2-乙烷磺酸)、MOPS(3-吗啉丙磺酸)、MES(2-(N-吗啡啉)乙磺酸)、三羟甲基甲胺-盐酸(Tris-HCl)、三羟甲基甲胺-硼酸(Tris-Boric acid)、三羟甲基甲胺-乙酸(Tris-Acetic acid)中的一种或多种。The aqueous buffer solution is selected from the group consisting of acetic acid-sodium acetate, boric acid-borax, HEPES (N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid), MOPS (3-morpholine propanesulfonic acid) ), MES (2-(N-morpholine) ethanesulfonic acid), Tris-Hydrochloric acid (Tris-HCl), Tris-Boric acid, Tris-Boric acid One or more of Tris-Acetic acid.
使用水性缓冲溶液配制叶酸水溶液时，优选地，所述水性缓冲溶液选自三羟甲基甲胺-盐酸、三羟甲 基甲胺-硼酸、三羟甲基甲胺-乙酸中的一种或多种。When an aqueous buffer solution is used to prepare an aqueous folic acid solution, preferably, the aqueous buffer solution is selected from one of trimethylol methylamine-hydrochloric acid, trimethylol methylamine-boric acid, and trimethylol methylamine-acetic acid. Many kinds.
为了提高叶酸在水性缓冲液中的溶解性，所述水性缓冲溶液的pH值为7.0～9.5。In order to improve the solubility of folic acid in the aqueous buffer solution, the pH value of the aqueous buffer solution is 7.0-9.5.
在步骤(1)制得的叶酸水溶液中，所述叶酸的浓度为0.5mM-200mM，优选为5mM-100mM。In the aqueous folic acid solution prepared in step (1), the concentration of the folic acid is 0.5mM-200mM, preferably 5mM-100mM.
进一步地，在步骤(2)中，使用与步骤(1)相同的水性溶液配制金属离子水溶液。Further, in step (2), the same aqueous solution as in step (1) is used to prepare a metal ion aqueous solution.
所述金属离子选自一价或二价态时化学性质稳定的金属离子，优选地，所述金属离子选自钠、钾、锂、铷、铯、锌(Zn 2+)，铜(Cu 2+)，镍(Ni 2+)，钴(Co 2+)，锰(Mn 2+)，铬(Cr 3+)，钼(Mo 2+)，银(Ag +)，镉(Cd 2+)，镁(Mg 2+)，钙(Ca 2+)，铅(Pb 2+)、钡等金属中的一种或多种。 The metal ions are selected from metal ions that are chemically stable in a monovalent or divalent state. Preferably, the metal ions are selected from sodium, potassium, lithium, rubidium, cesium, zinc (Zn 2+ ), copper (Cu 2 + ), nickel (Ni 2+ ), cobalt (Co 2+ ), manganese (Mn 2+ ), chromium (Cr 3+ ), molybdenum (Mo 2+ ), silver (Ag + ), cadmium (Cd 2+ ) , One or more of magnesium (Mg 2+ ), calcium (Ca 2+ ), lead (Pb 2+ ), barium and other metals.
进一步地，所述金属离子包括锌、铜、镍、钴、锰、铬、钼、银、镉、镁、钙和铅中的一种或多种，优选所述金属离子包括锌、银、镁和钙离子中的一种或多种。Further, the metal ion includes one or more of zinc, copper, nickel, cobalt, manganese, chromium, molybdenum, silver, cadmium, magnesium, calcium and lead, preferably the metal ion includes zinc, silver, magnesium And one or more of calcium ions.
配制时，优选使用含有上述金属离子的无机盐进行配制，例如盐酸盐、硝酸盐或硫酸盐等，优选使用硝酸盐或硫酸盐进行配制，这是因为，大多数硝酸盐或硫酸盐的溶解性比较好，更容易配制金属离子水溶液。When preparing, it is preferable to use inorganic salts containing the above-mentioned metal ions, such as hydrochloride, nitrate or sulfate, etc., preferably to use nitrate or sulfate for preparation, because most of the nitrate or sulfate dissolves It has better performance and is easier to prepare metal ion aqueous solution.
在步骤(2)制得的金属离子水溶液中，所述金属离子的浓度为0.5mM-500mM，优选为5mM-400mM。In the metal ion aqueous solution prepared in step (2), the concentration of the metal ion is 0.5mM-500mM, preferably 5mM-400mM.
进一步地，在步骤(3)中，将叶酸水溶液和金属离子水溶液混合均匀，混合方法并不做具体限定，只要能够混合均匀即可，例如涡旋、搅拌或者超声等。Further, in step (3), the folic acid aqueous solution and the metal ion aqueous solution are uniformly mixed, and the mixing method is not specifically limited, as long as the mixing can be uniform, such as vortexing, stirring, or ultrasound.
混合后，所述叶酸在混合体系中的最终浓度为0.5mM-200mM，优选为5mM-100mM，更优选为5mM-50mM，例如15mM。After mixing, the final concentration of the folic acid in the mixing system is 0.5mM-200mM, preferably 5mM-100mM, more preferably 5mM-50mM, for example 15mM.
混合后，所述金属离子在混合体系中的最终浓度为0.5mM-500mM，优选为5mM-400mM，更优选为10mM-200mM，例如27mM。After mixing, the final concentration of the metal ions in the mixing system is 0.5mM-500mM, preferably 5mM-400mM, more preferably 10mM-200mM, for example 27mM.
研究发现，提高叶酸浓度可以使制得的溶液粘度或者水凝胶的机械强度(模量)显著提高；而叶酸浓度过低时无法制得水凝胶。Studies have found that increasing the concentration of folic acid can significantly increase the viscosity of the prepared solution or the mechanical strength (modulus) of the hydrogel; however, when the concentration of folic acid is too low, the hydrogel cannot be prepared.
步骤(3)中，叶酸水溶液和金属离子水溶液混合后，叶酸和金属离子的摩尔浓度比为1：0.1-10。In step (3), after the folic acid aqueous solution and the metal ion aqueous solution are mixed, the molar concentration ratio of folic acid and metal ion is 1:0.1-10.
为了进一步优化水凝胶的机械强度，使制得的水凝胶具有良好的机械强度且适合皮肤涂抹，优选地，混合后混合体系中叶酸和金属离子的摩尔比为1：0.4-10，优选为1:1-5。In order to further optimize the mechanical strength of the hydrogel so that the prepared hydrogel has good mechanical strength and is suitable for skin application, preferably, the molar ratio of folic acid and metal ions in the mixed system after mixing is 1:0.4-10, preferably It is 1:1-5.
叶酸水溶液和金属离子水溶液混合后，混合体系中叶酸和金属离子的用量比例会影响叶酸-金属离子水凝胶的流体性状。本发明中，所述叶酸-金属离子水凝胶具有溶液、凝胶或悬浮液等流体状态。After the folic acid aqueous solution and the metal ion aqueous solution are mixed, the dosage ratio of folic acid and metal ions in the mixed system will affect the fluid properties of the folic acid-metal ion hydrogel. In the present invention, the folic acid-metal ion hydrogel has a fluid state such as a solution, a gel, or a suspension.
具体地，在一种优选的实施方式中，叶酸-锌离子水凝胶的相图如图20所示：随着叶酸浓度的提高，叶酸-锌离子水凝胶形成凝胶态时的叶酸/锌离子摩尔比下限逐渐降低，上限逐渐提高，凝胶区域扩大；相应地，随着叶酸浓度的降低，叶酸-锌离子水凝胶形成凝胶态时的叶酸/锌离子摩尔比下限逐渐提高，上限逐渐降低，凝胶区域缩小。由图20还可知，在叶酸浓度不变的条件下，叶酸/锌离子摩尔比逐渐增大时，叶酸-锌离子水凝胶由溶液状态变为凝胶状态，再变成混悬液状态。Specifically, in a preferred embodiment, the phase diagram of the folic acid-zinc ion hydrogel is shown in Figure 20: as the concentration of folic acid increases, the folic acid/zinc ion hydrogel forms a gel state. The lower limit of the zinc ion molar ratio gradually decreases, the upper limit gradually increases, and the gel area expands; accordingly, as the concentration of folic acid decreases, the lower limit of the folate/zinc ion molar ratio when the folate-zinc ion hydrogel forms a gel state gradually increases. The upper limit gradually decreases and the gel area shrinks. It can also be seen from FIG. 20 that under the condition that the folate concentration is unchanged, when the folate/zinc ion molar ratio gradually increases, the folate-zinc ion hydrogel changes from a solution state to a gel state, and then to a suspension state.
优选地，叶酸浓度为15mM时，将叶酸水溶液和锌离子水溶液混合后，叶酸/锌离子摩尔比为1:0-0.9时，得到的水凝胶为溶液态；叶酸和锌离子的摩尔浓度比为1:0.9-1.9时，得到的水凝胶形态为凝胶态；叶酸和锌离子的摩尔浓度比为1:2.0-10优选为1:2.0-2.2时，得到的水凝胶形态为悬浊液，Preferably, when the folic acid concentration is 15mM, after mixing the folic acid aqueous solution and the zinc ion aqueous solution, the folic acid/zinc ion molar ratio is 1:0-0.9, the obtained hydrogel is in a solution state; the molar concentration ratio of folic acid and zinc ion When it is 1:0.9-1.9, the obtained hydrogel form is gel state; when the molar concentration ratio of folic acid and zinc ion is 1:2.0-10, preferably 1:2.0-2.2, the obtained hydrogel form is suspended Turbid liquid,
优选地，当所选用金属离子为锌离子、且叶酸浓度为10mM时，叶酸和锌离子的摩尔浓度比为1:0-1.0时，得到的水凝胶为溶液态；叶酸和锌离子的摩尔浓度比为1:1.0-1.9时，得到的水凝胶形态为凝胶态；叶酸和锌离子的摩尔浓度比为1:2.0-10优选为1:2.0-2.2时，得到的水凝胶形态为悬浊液。Preferably, when the selected metal ion is zinc ion and the concentration of folic acid is 10mM, and the molar concentration ratio of folic acid and zinc ion is 1:0-1.0, the obtained hydrogel is in a solution state; the molar concentration of folic acid and zinc ion When the ratio is 1:1.0-1.9, the obtained hydrogel form is gel state; when the molar concentration ratio of folic acid and zinc ion is 1:2.0-10, preferably 1:2.0-2.2, the obtained hydrogel form is Suspension.
优选地，当所选用金属离子为锌离子、且叶酸浓度为2mM时，叶酸和锌离子的摩尔浓度比为1:0-1.5时，得到的水凝胶为溶液态；叶酸和锌离子的摩尔浓度比为1:1.5-1.7时，得到的水凝胶形态为凝胶态；叶酸和锌离子的摩尔浓度比为1:1.9-10优选为1:2.0-2.2时，得到的水凝胶形态为悬浊液。Preferably, when the selected metal ion is zinc ion and the concentration of folic acid is 2mM, and the molar concentration ratio of folic acid and zinc ion is 1:0-1.5, the obtained hydrogel is in solution; the molar concentration of folic acid and zinc ion When the ratio is 1:1.5-1.7, the obtained hydrogel form is gel state; when the molar concentration ratio of folic acid and zinc ion is 1:1.9-10, preferably 1:2.0-2.2, the obtained hydrogel form is Suspension.
进一步研究发现，混合体系的pH值可能会影响水凝胶的成型和性质，混合体系碱性过强时制得的水凝胶的机械强度会显著减弱；碱性过低甚至为酸性时混合体系中发生沉淀而无法形成水凝胶。Further studies have found that the pH value of the mixed system may affect the formation and properties of the hydrogel. When the mixed system is too alkaline, the mechanical strength of the hydrogel will be significantly weakened; when the alkaline is too low or even acidic, the mixed system Precipitation occurred in the water and the hydrogel could not be formed.
可选地，调节混合体系溶液pH至中性或碱性，优选为7.0-9.5。Optionally, adjust the pH of the mixed system solution to neutral or alkaline, preferably 7.0-9.5.
将混合体系静置即可使金属离子和叶酸充分交联配位，形成水凝胶分子。The mixed system can be allowed to fully crosslink and coordinate with folic acid to form hydrogel molecules.
研究发现，基于叶酸的金属离子水凝胶(叶酸-金属离子凝胶)的形成是一个多步骤的过程：首先，叶酸的喋呤头基通过氢键作用形成了四联体，如图3所示，所述四联体之间进一步π-π堆积，形成纤维，如图4所示；最后，金属离子和叶酸的羧酸发生配位，将纤维交联，纤维相互缠绕形成网状结构的水凝胶分子，如图4所示。Studies have found that the formation of folic acid-based metal ion hydrogels (folate-metal ion gels) is a multi-step process: first, the pterin head group of folic acid forms a quadruplex through hydrogen bonding, as shown in Figure 3. As shown, the tetrads are further stacked by π-π to form fibers, as shown in Figure 4; finally, the metal ions and the carboxylic acid of folic acid are coordinated to crosslink the fibers, and the fibers are entangled to form a network structure. Hydrogel molecules, as shown in Figure 4.
当金属离子和叶酸分子形成的网状结构的水凝胶分子均匀分散在水中时形成水凝胶的溶液；当网状结构的水凝胶分子分散性良好并能够将水包在其中，使其丧失流动性时，形成凝胶状态；当该网状结构的水凝胶分子无法分散于水中时，形成了叶酸-金属离子水凝胶的混悬液。When the hydrogel molecules of the network structure formed by metal ions and folic acid molecules are uniformly dispersed in water, a hydrogel solution is formed; when the hydrogel molecules of the network structure have good dispersibility and can enclose water in it, When the fluidity is lost, a gel state is formed; when the hydrogel molecules of the network structure cannot be dispersed in water, a folic acid-metal ion hydrogel suspension is formed.
混合体系最好在室温下静置，所述室温是指10-35℃，优选为15-30℃，最好为20-30℃，例如25℃。The mixed system is best left to stand at room temperature. The room temperature refers to 10-35°C, preferably 15-30°C, and most preferably 20-30°C, for example 25°C.
步骤(3)中，静置形成水凝胶的时间与选用的金属离子的种类相关；优选静置1-2天，可以使水凝胶中交联分子更为稳定。In step (3), the time for standing to form the hydrogel is related to the type of metal ion selected; preferably, standing for 1-2 days can make the crosslinking molecules in the hydrogel more stable.
在本发明提供的制备方法中，通过将金属离子事先制备为水溶液，可以避免金属离子固体与叶酸水溶液成胶的不均匀性，制得更为均匀可靠的叶酸-金属离子水凝胶。In the preparation method provided by the present invention, by preparing the metal ion as an aqueous solution in advance, the inhomogeneity of the gelation between the metal ion solid and the folic acid aqueous solution can be avoided, and a more uniform and reliable folic acid-metal ion hydrogel can be prepared.
本发明还提供了一种上述方法制得的叶酸-金属离子水凝胶，所述水凝胶具有网状结构，包括配位连接的金属离子和叶酸分子。其中，叶酸的喋呤头基通过氢键形成四联体，四联体之间π-π堆积形成纤维，金属离子和叶酸的羧酸发生配位，使纤维交联并相互缠绕形成网状结构。The present invention also provides a folic acid-metal ion hydrogel prepared by the above method. The hydrogel has a network structure and includes coordinately connected metal ions and folic acid molecules. Among them, the pterin head group of folic acid forms a quadruplex through hydrogen bonding, and the π-π stacks between the tetrads to form fibers. The metal ions and the carboxylic acid of folic acid coordinate, so that the fibers are cross-linked and intertwined to form a network structure. .
其中，在所述水凝胶的XRD图谱中，在约8.5°和约26.5°处存在特征峰；约8.5°处特征峰对应的是单根纤维直径，约26.5°处特征峰对应的是π-π堆积的距离。Wherein, in the XRD pattern of the hydrogel, there are characteristic peaks at about 8.5° and about 26.5°; the characteristic peak at about 8.5° corresponds to the diameter of a single fiber, and the characteristic peak at about 26.5° corresponds to π- π stacking distance.
所述水凝胶在圆二色谱的谱图中存在叶酸的手性堆积峰；特别地，在300nm附近和260nm附近显示手性信号。The hydrogel has a chiral stacking peak of folic acid in the spectrum of the circular dichroism; in particular, it shows chiral signals near 300 nm and 260 nm.
优选地，当所述水凝胶为叶酸-锌水凝胶时，在300nm附近显示负信号，在260nm附近显示正信号。Preferably, when the hydrogel is folic acid-zinc hydrogel, it shows a negative signal around 300 nm and a positive signal around 260 nm.
上述方法制得的叶酸-金属离子水凝胶具有良好的生物相容性，可以作为生物材料应用于生物医学领域。The folic acid-metal ion hydrogel prepared by the above method has good biocompatibility and can be used as a biological material in the field of biomedicine.
在一种优选的实施方式中，所述叶酸-金属离子水凝胶可用于细胞三维培养，更优选作为细胞培养基质(或支撑材料)用于细胞培养。In a preferred embodiment, the folate-metal ion hydrogel can be used for three-dimensional cell culture, more preferably as a cell culture substrate (or support material) for cell culture.
在一种优选的实施方式中，所述叶酸-金属离子水凝胶可以用于生物三维打印。In a preferred embodiment, the folate-metal ion hydrogel can be used for three-dimensional biological printing.
在一种优选的实施方式中，所述叶酸-金属离子水凝胶可以作为营养补充剂，同时提供叶酸和微量金属元素。In a preferred embodiment, the folic acid-metal ion hydrogel can be used as a nutritional supplement while providing folic acid and trace metal elements.
进一步地，研究发现，将叶酸制备为叶酸-金属离子水凝胶后依旧具有和叶酸类似的紫外吸收的性质，具有在UV-A和UV-B区域吸收紫外作用，可以作为紫外吸收剂或防晒剂用于防晒，使得叶酸-金属离子水凝胶具有在防晒中的用途。Further, research has found that the preparation of folic acid as a folic acid-metal ion hydrogel still has ultraviolet absorption properties similar to folic acid, has the effect of absorbing ultraviolet in the UV-A and UV-B regions, and can be used as an ultraviolet absorber or sunscreen The agent is used for sun protection, so that the folic acid-metal ion hydrogel has the use in sun protection.
将叶酸制备为叶酸-金属离子水凝胶后具有凝胶的粘稠性、涂展性和均匀性，非常有利于防晒剂的制备和成型。叶酸-金属离子水凝胶类似市售的芦荟胶，相比于传统的油腻性防晒霜，水凝胶的消费者体验明显提高，可以提高消费者的接受程度。After folic acid is prepared into folic acid-metal ion hydrogel, it has the viscosity, spreadability and uniformity of the gel, which is very beneficial to the preparation and molding of sunscreen. Folic acid-metal ion hydrogel is similar to commercially available aloe vera gel. Compared with traditional greasy sunscreens, the hydrogel has a significantly improved consumer experience and can increase consumer acceptance.
将叶酸加入金属离子制备成叶酸-金属离子水凝胶后，其防晒效果几乎保持恒定，而且不同比例的叶酸-金属离子可以得到粘稠度不同甚至流动状态(或流体状态)也不同的水凝胶，为生产带来多样性的同时也为消费者提供了多种选择。After folic acid is added to metal ions to prepare folic acid-metal ion hydrogel, the sunscreen effect remains almost constant, and different ratios of folic acid-metal ions can get different viscosity and even different fluid state (or fluid state). Glue, while bringing diversity to production, also provides consumers with multiple choices.
在一种更优选的实施方式中，所述叶酸-金属离子水凝胶还可以与氧化锌复配形成防晒剂，氧化锌具有物理防晒的作用，而叶酸-金属离子水凝胶具有化学防晒即紫外吸收剂的作用，通过二者的复配，显著提高防晒效果。In a more preferred embodiment, the folic acid-metal ion hydrogel can also be compounded with zinc oxide to form a sunscreen. Zinc oxide has a physical sunscreen effect, and the folic acid-metal ion hydrogel has a chemical sunscreen effect. The role of ultraviolet absorber, through the combination of the two, significantly improves the sun protection effect.
所述氧化锌在防晒剂中的质量分数为0.1％-10％，优选为0.5％-5％。The mass fraction of the zinc oxide in the sunscreen is 0.1%-10%, preferably 0.5%-5%.
更优选所述氧化锌在叶酸-金属离子水凝胶和氧化锌复配混合物中的质量分数为0.1％-10％，优选为0.5％-5％。More preferably, the mass fraction of the zinc oxide in the folic acid-metal ion hydrogel and zinc oxide compound mixture is 0.1%-10%, preferably 0.5%-5%.
叶酸-金属离子水凝胶和氧化锌复配时，将设定用量的二者混合均匀后即可。When folic acid-metal ion hydrogel and zinc oxide are compounded, just mix the two in the set dosage evenly.
较好地，叶酸-金属离子水凝胶和氧化锌复配时，任选地，还可以加入其它不同作用的组分，如皮肤营养剂、溶剂、增稠剂、络合剂和/或防晒物质。Preferably, when folic acid-metal ion hydrogel and zinc oxide are compounded, optionally, other components with different functions can be added, such as skin nutrients, solvents, thickeners, complexing agents and/or sunscreens. substance.
皮肤营养剂包括甘油、维生素E、单一或多种动植物提取物、尿囊素和葵基葡糖苷中的一种或多种；Skin nutrition agents include one or more of glycerin, vitamin E, single or multiple animal and plant extracts, allantoin and decyl glucoside;
溶剂包括水、丙二醇、乙醇、甘油硬脂酸酯、椰油酰肌氨酸钠、鲸蜡醇磷酸酯钾、鲸蜡醇、辛甘醇、十一烷、异十六烷、异十二烷、苯乙醇苯甲酸酯、碳酸二辛酯、二辛基醚、碳酸丙二醇酯、丙烯酰胺/丙烯酸铵共聚物和聚山梨醇酯-20中的一种或多种；Solvents include water, propylene glycol, ethanol, glyceryl stearate, sodium cocoyl sarcosinate, potassium cetyl phosphate, cetyl alcohol, octyl glycol, undecane, isohexadecane, isododecane , Phenethyl alcohol benzoate, dioctyl carbonate, dioctyl ether, propylene glycol carbonate, acrylamide/ammonium acrylate copolymer and polysorbate-20 one or more;
所述络合剂包括EDTA二钠；The complexing agent includes disodium EDTA;
所述增稠剂包括淀粉辛烯基琥珀酸铝、黄原胶、氧化铝、甘油三(乙基己酸)酯、三乙氧基辛基硅烷、氯化钠、滑石粉、氢氧化铝、二硬脂二甲铵锂蒙脱石、硅石和异硬脂酸中的一种或多种；The thickener includes aluminum starch octenyl succinate, xanthan gum, aluminum oxide, glycerol tris(ethylhexanoate), triethoxyoctyl silane, sodium chloride, talc, aluminum hydroxide, One or more of distearyl dimethyl ammonium hectorite, silica and isostearic acid;
所述防晒物质包括氧化锌、甲氧基肉桂酸乙基己酯、奥克立林、聚硅氧烷-15、二氧化钛、二乙氨羟苯甲酰基苯甲酸己酯、双-乙基己氧苯酚甲氧苯基三嗪、阿伏苯宗、二苯甲酮-3、水杨酸辛酯和甲氧基肉桂酸辛酯中的一种或多种。The sunscreen substance includes zinc oxide, ethylhexyl methoxycinnamate, octoclilin, polysiloxane-15, titanium dioxide, diethylcarbamyl hexyl benzoate, bis-ethylhexyloxy One or more of phenol methoxyphenyl triazine, avobenzone, benzophenone-3, octyl salicylate, and octyl methoxycinnamate.
任选地，还可以添加防腐剂，例如山梨酸或山梨酸钾。Optionally, preservatives such as sorbic acid or potassium sorbate can also be added.
所述叶酸-金属离子水凝胶制备为防晒剂时，优选制备为常用的膏剂、霜剂或凝胶剂，涂抹于皮肤时用量为0.1-10mg/cm 2，具有较好的防紫外线作用。 When the folic acid-metal ion hydrogel is prepared as a sunscreen, it is preferably prepared as a commonly used ointment, cream or gel, and the amount when applied to the skin is 0.1-10 mg/cm 2 , which has a good UV protection effect.
本发明提供的叶酸-金属离子水凝胶制备简单、工艺可控可调，具有较好的紫外吸收作用，能够与其他化学吸收剂和物理防晒剂复配使用，提高防紫外线和防晒的效果。所述叶酸-金属离子水凝胶安全性好可靠性高，化学性质稳定，制备为防晒剂后非常清爽，使用体验良好。The folic acid-metal ion hydrogel provided by the invention is simple to prepare, has a controllable and adjustable process, has good ultraviolet absorption, can be used in combination with other chemical absorbents and physical sunscreens, and improves the effects of ultraviolet protection and sun protection. The folic acid-metal ion hydrogel has good safety, high reliability, stable chemical properties, is very refreshing after being prepared as a sunscreen, and has a good user experience.
实施例Example
实施例1Example 1
在2mL试管中称取叶酸10.74mg，加入超纯水0.225ml，加入氢氧化钾调节溶液pH至9左右，得到叶 酸水溶液。Weigh 10.74 mg of folic acid in a 2 mL test tube, add 0.225 ml of ultrapure water, and add potassium hydroxide to adjust the pH of the solution to about 9, to obtain an aqueous folic acid solution.
另取试管，加入硝酸锌10.04mg，加入超纯水1.275ml，得到金属离子水溶液。Take another test tube, add 10.04mg of zinc nitrate, add 1.275ml of ultrapure water to obtain a metal ion aqueous solution.
将金属离子水溶液加入叶酸水溶液中，使混合溶液中叶酸的浓度为15mM、锌离子浓度为22.5mM。涡旋搅拌混匀后静置，混合溶液最初呈现浑浊，经过10分钟后逐渐澄清透明，得到物质的量浓度比为1-1.5(叶酸:锌离子＝1：1.5，下同)的叶酸-锌水凝胶，结果如图1所示。The metal ion aqueous solution was added to the folic acid aqueous solution so that the folic acid concentration in the mixed solution was 15 mM and the zinc ion concentration was 22.5 mM. After vortexing and mixing, the mixed solution was initially turbid, and gradually became clear and transparent after 10 minutes. The folic acid-zinc with a substance concentration ratio of 1-1.5 (folic acid: zinc ion = 1: 1.5, the same below) was obtained. Hydrogel, the result is shown in Figure 1.
实施例2Example 2
配制三羟甲基甲胺-硼酸缓冲溶液，pH控制在9左右。在2mL试管中称取叶酸10.74mg，加入三羟甲基甲胺-硼酸0.225ml，得到叶酸水溶液。Prepare the trimethylolmethylamine-boric acid buffer solution, and control the pH at around 9. Weigh 10.74 mg of folic acid in a 2 mL test tube, add 0.225 ml of trimethylol methylamine-boric acid to obtain an aqueous folic acid solution.
另取试管，加入硝酸铜8.154mg，加入三羟甲基甲胺-硼酸1.275ml，得到金属离子水溶液。Take another test tube, add 8.154 mg of copper nitrate, and add 1.275 ml of trimethylol methylamine-boric acid to obtain a metal ion aqueous solution.
将金属离子水溶液加入叶酸水溶液中，使混合溶液中叶酸的浓度为15mM、铜离子浓度为22.5mM。涡旋搅拌混匀后静置，混合溶液最初呈现浑浊，经过两天后逐渐澄清透明，得到叶酸-铜水凝胶，结果如图1所示。The metal ion aqueous solution was added to the folic acid aqueous solution so that the folic acid concentration in the mixed solution was 15 mM and the copper ion concentration was 22.5 mM. After vortexing and mixing, the mixed solution was initially turbid, and gradually became clear and transparent after two days, and the folic acid-copper hydrogel was obtained. The result is shown in Figure 1.
实施例3Example 3
在与实施例2基本相同的条件下，分别将金属离子换为Ni 2+、Mg 2+、Pb 2+、Ag +，分别制备的叶酸-镍水凝胶、叶酸-镁水凝胶、叶酸-铅水凝胶、叶酸-银水凝胶，结果如图1所示。 Under basically the same conditions as in Example 2, the metal ions were replaced with Ni 2+ , Mg 2+ , Pb 2+ , Ag + , respectively, and folic acid-nickel hydrogel, folic acid-magnesium hydrogel, and folic acid were prepared separately -Lead hydrogel, folic acid-silver hydrogel, the results are shown in Figure 1.
实施例4Example 4
将叶酸加入超纯水中，加入氢氧化钾调节溶液pH至9左右使叶酸溶解，得到叶酸水溶液。Folic acid is added to ultrapure water, potassium hydroxide is added to adjust the pH of the solution to about 9 to dissolve the folic acid to obtain an aqueous folic acid solution.
将硝酸锌加入超纯水中溶解，得到金属离子水溶液。Zinc nitrate is added to ultra-pure water to dissolve to obtain a metal ion aqueous solution.
分别取金属离子水溶液和叶酸水溶液并混合，使混合溶液中叶酸的最终质量分数为0.72wt％；锌离子最终质量分数为0.90wt％。涡旋搅拌混匀后静置，混合溶液最初呈现浑浊，经过10分钟后逐渐澄清透明，得到叶酸-锌离子(F-Zn 2+)物质的量浓度比为1-1.8(叶酸:锌离子＝1：1.8，下同)的叶酸-锌水凝胶。 The metal ion aqueous solution and the folic acid aqueous solution are taken and mixed separately, so that the final mass fraction of folic acid in the mixed solution is 0.72 wt%; the final mass fraction of zinc ion is 0.90 wt%. After vortexing and mixing, the mixed solution was initially turbid, and gradually became clear and transparent after 10 minutes. The amount and concentration ratio of folic acid-zinc ion (F-Zn 2+ ) was 1-1.8 (folate: zinc ion = 1:1.8, the same below) folate-zinc hydrogel.
实施例5Example 5
制备方法同实施例4相似，区别在于：The preparation method is similar to Example 4, the difference is:
混合溶液中叶酸的最终质量分数为0.72wt％；锌离子的最终质量分数为0.28wt％。涡旋搅拌混匀后静置10分钟得到叶酸-锌离子(F-Zn 2+)物质的量浓度比为1-0.4的叶酸-锌水凝胶，此时为溶液状态。 The final mass fraction of folic acid in the mixed solution is 0.72 wt%; the final mass fraction of zinc ions is 0.28 wt%. Vortex, stir and mix, and then stand for 10 minutes to obtain a folic acid-zinc hydrogel with a folic acid-zinc ion (F-Zn 2+ ) substance concentration ratio of 1-0.4, which is now in a solution state.
实施例6Example 6
制备方法同实施例4相似，区别在于：The preparation method is similar to Example 4, the difference is:
混合溶液中叶酸的最终质量分数为0.72wt％；锌离子的最终质量分数为0.46wt％。涡旋搅拌混匀后静置10分钟得到叶酸-锌离子(F-Zn 2+)物质的量浓度比为1-0.8的叶酸-锌水凝胶，此时为溶液状态。 The final mass fraction of folic acid in the mixed solution is 0.72 wt%; the final mass fraction of zinc ions is 0.46 wt%. Vortex, stir and mix, and then stand for 10 minutes to obtain a folic acid-zinc hydrogel with a folic acid-zinc ion (F-Zn 2+ ) concentration ratio of 1-0.8, which is now in a solution state.
实施例7Example 7
制备方法同实施例4相似，区别在于：The preparation method is similar to Example 4, the difference is:
混合溶液中叶酸的最终质量分数为0.72wt％；锌离子的最终质量分数为0.63wt％。涡旋搅拌混匀后静置10分钟得到叶酸-锌离子(F-Zn 2+)物质的量浓度比为1-1.2的叶酸-锌水凝胶，此时为凝胶状态。 The final mass fraction of folic acid in the mixed solution is 0.72 wt%; the final mass fraction of zinc ions is 0.63 wt%. Vortex, stir and mix, and then stand for 10 minutes to obtain a folic acid-zinc hydrogel with a folic acid-zinc ion (F-Zn 2+ ) substance concentration ratio of 1-1.2, which is now in a gel state.
实施例8Example 8
制备方法同实施例4相似，区别在于：The preparation method is similar to Example 4, the difference is:
混合溶液中叶酸的最终质量分数为0.72wt％；锌离子的最终质量分数为0.81wt％。涡旋搅拌混匀后静置10分钟得到叶酸-锌离子(F-Zn 2+)物质的量浓度比为1-1.6的叶酸-锌水凝胶，此时为凝胶状态，该凝胶流变性非常好。 The final mass fraction of folic acid in the mixed solution is 0.72 wt%; the final mass fraction of zinc ions is 0.81 wt%. After vortexing and mixing, let it stand for 10 minutes to obtain a folic acid-zinc hydrogel with a folic acid-zinc ion (F-Zn 2+ ) concentration ratio of 1-1.6. At this time, it is in a gel state. Denaturation is very good.
实施例9Example 9
在与实施例1或实施例4相似的实验条件下，分别制备叶酸和锌离子摩尔浓度比为1-1.0，1-1.4，1-1.5，1-1.7，1-1.9的叶酸-锌水凝胶。Under similar experimental conditions as in Example 1 or Example 4, folic acid-zinc hydrogels with molar concentration ratios of folic acid and zinc ions of 1-1.0, 1-1.4, 1-1.5, 1-1.7, 1-1.9 were prepared, respectively. glue.
实施例10Example 10
配制三羟甲基甲胺-硼酸缓冲溶液，pH控制在9.5左右。称取叶酸10.74mg溶于三羟甲基甲胺-硼酸缓冲溶液0.225mL，得到叶酸水溶液。Prepare the trimethylol methylamine-boric acid buffer solution, and control the pH at about 9.5. Weigh 10.74 mg of folic acid and dissolve in 0.225 mL of trimethylol methylamine-boric acid buffer solution to obtain an aqueous folic acid solution.
另取硝酸银19.38mg，溶于三羟甲基甲胺-硼酸缓冲溶液1.275mL，得到金属离子水溶液。Another 19.38 mg of silver nitrate was dissolved in 1.275 mL of trimethylolmethylamine-boric acid buffer solution to obtain a metal ion aqueous solution.
分别取上述金属离子水溶液和叶酸水溶液并混合，最终混合溶液中叶酸的质量分数为0.72wt％；银离子的质量分数为0.89wt％。涡旋搅拌混匀后静置，混合溶液最初呈现浑浊，经过30分钟后逐渐澄清透明，得到叶酸和银离子物质的量浓度比为1：5的叶酸-银水凝胶。The metal ion aqueous solution and the folic acid aqueous solution are taken and mixed separately, and the mass fraction of folic acid in the final mixed solution is 0.72 wt%; the mass fraction of silver ion is 0.89 wt%. After vortexing and mixing, the mixed solution was initially turbid, and gradually became clear and transparent after 30 minutes, and a folic acid-silver hydrogel with a concentration ratio of folic acid and silver ions of 1:5 was obtained.
实施例11Example 11
制备方法同实施例10相似，区别在于：The preparation method is similar to that of Example 10, except that:
混合溶液中叶酸的最终质量分数为0.72wt％；钙离子的最终质量分数为0.37wt％。涡旋搅拌混匀后静置1小时得到叶酸和钙离子物质的量浓度比为1：1.5的叶酸-钙水凝胶。The final mass fraction of folic acid in the mixed solution is 0.72 wt%; the final mass fraction of calcium ions is 0.37 wt%. After vortexing and mixing, let it stand for 1 hour to obtain a folic acid-calcium hydrogel with a folic acid and calcium ion substance concentration ratio of 1:1.5.
实施例12Example 12
制备方法同实施例10基本相同，区别在于：The preparation method is basically the same as that of Example 10, the difference lies in:
混合溶液中叶酸的最终质量分数为0.72wt％；镁离子的最终质量分数为0.33wt％。涡旋搅拌混匀后静置1小时得到叶酸和镁离子物质的量浓度比为1：1.5的叶酸-镁水凝胶。The final mass fraction of folic acid in the mixed solution is 0.72 wt%; the final mass fraction of magnesium ions is 0.33 wt%. Vortex, stir and mix, and then stand for 1 hour to obtain a folic acid-magnesium hydrogel with a folic acid and magnesium ion substance concentration ratio of 1:1.5.
实验例Experimental example
实验例1三维打印Experimental example 1 3D printing
将实施例1制备的叶酸-锌水凝胶注入注射针管中，将水凝胶从针尖中注出至塑料板上，在注出的同时移动针尖的空间位置，即可得到多种多样的水凝胶结构。结果如图2所示，图2中(a)为打印的简易装置注射针管，(b)为打印出的含有数字和字母的水凝胶结构；(c)和(e)为打印出的空心三角形的水凝胶结构；(d)为打印出的金字塔形的水凝胶结构；(g)为打印出的心形水凝胶结构；(f)为打印出的四边形水凝胶结构。Inject the folic acid-zinc hydrogel prepared in Example 1 into the injection needle, inject the hydrogel from the needle tip onto the plastic plate, move the space position of the needle tip while injecting, and then obtain a variety of water Gel structure. The result is shown in Figure 2. In Figure 2, (a) is the printed simple device injection needle, (b) is the printed hydrogel structure containing numbers and letters; (c) and (e) are the printed hollow Triangular hydrogel structure; (d) is the printed pyramid-shaped hydrogel structure; (g) is the printed heart-shaped hydrogel structure; (f) is the printed quadrilateral hydrogel structure.
实验例2细胞培养和打印Experimental example 2 Cell culture and printing
选用HeLa细胞和hRPE细胞进行实验。培养条件是Dulbecco’s modified Eagle’s medium(DMEM，高糖培养基)+10％牛血清蛋白+1％双抗，培养环境为5％CO 2，37℃。 Choose HeLa cells and hRPE cells for experiments. The culture condition is Dulbecco's modified Eagle's medium (DMEM, high glucose medium) + 10% bovine serum protein + 1% double antibody, and the culture environment is 5% CO 2 , 37°C.
细胞毒性试验使用的是CCK-8染色法，HeLa或hRPE细胞在96孔板中培养，细胞浓度约为5×10 3细胞/mL；在培养基中加入实施例1制得的叶酸-锌水凝胶，浓度分别为0、2、4、6、8、10mg/mL。培养一段时间后，使用CCK-8染色，根据450nm处的吸光值确定细胞存活率(cell viability)。 The cytotoxicity test used the CCK-8 staining method. HeLa or hRPE cells were cultured in 96-well plates at a cell concentration of about 5×10 3 cells/mL; the folic acid-zinc water prepared in Example 1 was added to the medium Gels with concentrations of 0, 2, 4, 6, 8, and 10 mg/mL. After culturing for a period of time, use CCK-8 staining to determine cell viability based on the absorbance at 450 nm.
如图11和图12所示，叶酸-锌水凝胶在浓度为0-10mg/mL范围内，hRPE和HeLa细胞的存活率均大于97％，表明叶酸-锌水凝胶对于hRPE和HeLa细胞几乎没有毒性。As shown in Figure 11 and Figure 12, the survival rate of folic acid-zinc hydrogel in the concentration range of 0-10 mg/mL is greater than 97% for hRPE and HeLa cells, indicating that folic acid-zinc hydrogel is effective for hRPE and HeLa cells. There is almost no toxicity.
细胞打印的过程如下：The process of cell printing is as follows:
按照实施例1的方法制备叶酸-锌水凝胶，将悬浮的HeLa或者hRPE细胞在最后混合时加入，细胞浓度为1×10 6细胞/mL,加入量为0.2mL，保证均匀混合。通过移液枪的枪头将水凝胶打印至塑料皿上，将该含有细胞的水凝胶浸没在培养基中培养。三维培养的细胞使用Calcein AM(钙黄绿素)和Ethidium homodimer-1(乙锭同二聚体-1)染色，激光共聚焦显微镜三维扫描，确定凝胶中的细胞生命状态和存活情况。 The folic acid-zinc hydrogel was prepared according to the method of Example 1, and the suspended HeLa or hRPE cells were added at the final mixing. The cell concentration was 1×10 6 cells/mL, and the addition amount was 0.2 mL to ensure uniform mixing. The hydrogel is printed on a plastic dish with the tip of a pipette gun, and the hydrogel containing cells is immersed in the culture medium for culture. Three-dimensionally cultured cells were stained with Calcein AM (calcein) and Ethidium homodimer-1 (ethidium homodimer-1), and three-dimensional scanning with a laser confocal microscope to determine the life state and survival of the cells in the gel.
结果如图13和图14所示，显示HeLa和hRPE的存活率均大于98.5％。这一结果说明叶酸-锌水凝胶可以作为三维培养的基质，维持hRPE和HeLa细胞的正常存活和生命活动。The results are shown in Figure 13 and Figure 14, showing that the survival rates of HeLa and hRPE are both greater than 98.5%. This result indicates that folic acid-zinc hydrogel can be used as a three-dimensional culture matrix to maintain the normal survival and life activities of hRPE and HeLa cells.
实验例3流变表征Experimental example 3 rheological characterization
对实施例4和实施例9制得的1-1.7、1-1.8和1-1.9叶酸-锌水凝胶进行流变学测试，图5和图6示出了其流变学测试结果。Rheology tests were performed on the 1-1.7, 1-1.8, and 1-1.9 folate-zinc hydrogels prepared in Example 4 and Example 9. Figs. 5 and 6 show the rheological test results.
图5为振幅扫描，不同比例的叶酸-锌水凝胶的储能模量G’和耗能模量G”在达到屈服值前均保持稳定，且G’比G”大一个数量级。Figure 5 shows the amplitude sweep. The storage modulus G'and the energy dissipation modulus G" of different ratios of folic acid-zinc hydrogels remain stable before reaching the yield value, and G'is an order of magnitude larger than G".
图6为频率扫描，不同比例的叶酸-锌水凝胶的频率在0.05Hz到5Hz的范围内，凝胶的储能模量G’和耗能模量G”均保持稳定。Figure 6 shows the frequency sweep. The frequency of different ratios of folic acid-zinc hydrogel is in the range of 0.05 Hz to 5 Hz, and the storage modulus G'and the energy dissipation modulus G" of the gel remain stable.
流变学测试结果表明了水凝胶体系独特的粘弹性性质，清晰地证明了在应力达到屈服值前凝胶保持典型的固态性质。另外，随着锌离子比例不断提高(从1:1.7到1:1.9)，凝胶的储能模量G’、耗能模量G”和屈服值都会提高，这说明金属离子的比例是调控水凝胶机械强度的有效方式。The rheological test results show the unique viscoelastic properties of the hydrogel system and clearly prove that the gel retains the typical solid state properties before the stress reaches the yield value. In addition, as the ratio of zinc ions continues to increase (from 1:1.7 to 1:1.9), the storage modulus G', energy dissipation modulus G" and yield value of the gel will increase, which shows that the ratio of metal ions is adjusted An effective way of mechanical strength of hydrogels.
实验例4微观形貌Experimental example 4 micro morphology
对实施例1得到的叶酸-锌水凝胶和实施例2得到的叶酸-铜水凝胶进行透射电子显微镜观察。The folic acid-zinc hydrogel obtained in Example 1 and the folic acid-copper hydrogel obtained in Example 2 were observed by a transmission electron microscope.
结果如图7所示，(a)为叶酸-锌水凝胶的透射电镜图，(b)叶酸-铜水凝胶的透射电镜图。可以发现，叶酸-锌水凝胶和叶酸-铜水凝胶均为纤维交联形成的网状结构。The results are shown in Figure 7, (a) is a transmission electron micrograph of folic acid-zinc hydrogel, (b) a transmission electron micrograph of folic acid-copper hydrogel. It can be found that both folic acid-zinc hydrogel and folic acid-copper hydrogel are network structures formed by fiber cross-linking.
同时，对叶酸-锌水凝胶和叶酸-铜水凝胶进行扫描电子显微镜观察。结果如图8所示，(a)为叶酸-锌水凝胶的扫描电镜图，(b)叶酸-铜水凝胶的扫描电镜图。同样地，可以发现，叶酸-锌水凝胶和叶酸-铜水凝胶均为纤维交联形成的网状结构。At the same time, folic acid-zinc hydrogel and folic acid-copper hydrogel were observed by scanning electron microscope. The results are shown in Figure 8, (a) is a scanning electron micrograph of folic acid-zinc hydrogel, (b) a scanning electron micrograph of folic acid-copper hydrogel. Similarly, it can be found that both folic acid-zinc hydrogel and folic acid-copper hydrogel are network structures formed by fiber cross-linking.
实验例5 XRD表征Experimental example 5 XRD characterization
对实施例1得到的叶酸-锌水凝胶和实施例2得到的叶酸-铜水凝胶进行XRD表征，以明确微观结构。The folic acid-zinc hydrogel obtained in Example 1 and the folic acid-copper hydrogel obtained in Example 2 were characterized by XRD to clarify the microstructure.
XRD结果如图9所示，叶酸-锌水凝胶和叶酸-铜水凝胶中均存在两个特征峰，一个是位于小角区的约8.5°峰，对应1nm距离，为单根纤维直径；另一个是约26.5°峰，对应0.34nm，是π-π堆积的距离。The XRD results are shown in Figure 9. There are two characteristic peaks in both folic acid-zinc hydrogel and folic acid-copper hydrogel, one is a peak at about 8.5° in the small corner area, corresponding to a distance of 1nm, which is a single fiber diameter; The other is about 26.5° peak, corresponding to 0.34nm, which is the distance of π-π stacking.
实验例6水凝胶的圆二色谱表征Experimental Example 6 Circular Dichroism Characterization of Hydrogel
对实施例1、实施例6～8、实施例9制得的不同比例的叶酸-锌水凝胶进行圆二色谱检测，叶酸水溶液(叶酸-Zn 2+1-0)做对照，结果如图10所示。 The folic acid-zinc hydrogels in different proportions prepared in Example 1, Examples 6-8 and Example 9 were detected by circular dichroism, and the folic acid aqueous solution (folic acid-Zn 2+ 1-0) was used as a control. The results are shown in the figure 10 shown.
可以发现：水凝胶在300nm附近显示很强的负信号，在260nm附近显示很强的正信号，说明叶酸在水凝胶中发生了手性的堆积，这也是水凝胶的一大特征。It can be found that the hydrogel shows a strong negative signal near 300nm and a strong positive signal near 260nm, indicating that the folic acid has been chiral accumulation in the hydrogel, which is also a major feature of the hydrogel.
另外，在实验摩尔比范围中，随着金属比例的不断提高，手性信号不断增强。In addition, in the experimental molar ratio range, as the metal ratio continues to increase, the chiral signal continues to increase.
实验例7不同化学吸收剂(防晒组分)的紫外吸收行为Experimental example 7 UV absorption behavior of different chemical absorbers (sunscreen components)
使用乙酸乙酯分别配制质量百分比为0.01％的阿伏苯宗、二苯甲酮-3、水杨酸辛酯和甲氧基肉桂酸辛酯溶液。按照实施例4的方法配制叶酸摩尔浓度为15mM的水溶液。对上述溶液在250-450nm范围内紫外 扫描，随后将吸光值对浓度和光程归一化为1％和1cm(根据朗伯比尔定律)，结果如图15所示，可以发现：Avobenzone, benzophenone-3, octyl salicylate and octyl methoxycinnamate solutions with a mass percentage of 0.01% were prepared using ethyl acetate. According to the method of Example 4, an aqueous solution with a molar concentration of folic acid of 15 mM was prepared. Scan the above solution with UV in the range of 250-450nm, and then normalize the absorbance value to the concentration and optical path to 1% and 1cm (according to Lambert Beer's law). The result is shown in Figure 15 and it can be found that:
在UV-B(280-320nm)区域内，叶酸的紫外吸收能力与二苯甲酮-3相当，高于水杨酸辛酯和阿伏苯宗，可与甲氧基肉桂酸辛酯的紫外吸收能力相比拟；In the UV-B (280-320nm) region, the UV absorption capacity of folic acid is equivalent to that of benzophenone-3, higher than that of octyl salicylate and avobenzone, and can be compared with the UV absorption capacity of octyl methoxycinnamate Compare
UV-AII(320-340nm)区域内，叶酸的紫外吸收能力高于水杨酸辛酯，与二苯甲酮-3和甲氧基肉桂酸辛酯的紫外吸收能力相比拟；In the UV-AII (320-340nm) region, the UV absorption capacity of folic acid is higher than that of octyl salicylate, which is comparable to the UV absorption capacity of benzophenone-3 and octyl methoxycinnamate;
UV-AI(340-400nm)区域内，叶酸的紫外吸收能力高于水杨酸辛酯、二苯甲酮-3和甲氧基肉桂酸辛酯，仅次于阿伏苯宗的紫外吸收能力。In the UV-AI (340-400nm) region, the UV absorption capacity of folic acid is higher than that of octyl salicylate, benzophenone-3 and octyl methoxycinnamate, and is second only to the UV absorption capacity of avobenzone.
实验例8不同叶酸-锌离子配比水凝胶的紫外吸收行为Experimental Example 8 UV absorption behavior of hydrogels with different folate-zinc ion ratios
实验例4中的叶酸水溶液与实施例5-8中制得的叶酸-锌水凝胶。由于叶酸-锌水凝胶具有良好的流动性，使用胶头滴管滴入比色皿中即可测量。The folic acid aqueous solution in Experimental Example 4 and the folic acid-zinc hydrogel prepared in Examples 5-8. Since folic acid-zinc hydrogel has good fluidity, it can be measured by dropping it into a cuvette with a plastic-tip dropper.
对上述溶液在250-450nm范围内紫外扫描，随后将吸光值对浓度和光程归一化为1％和1cm(根据朗伯比尔定律)，得到结果如图16所示，可以发现：Scan the above solution with UV in the range of 250-450nm, and then normalize the absorbance value to the concentration and optical path to 1% and 1cm (according to Lambert Beer's law). The result is shown in Figure 16, and it can be found that:
叶酸制备为水凝胶后，其紫外吸收行为与叶酸水溶液的紫外吸收行为基本相同，只有叶酸-锌离子(F-Zn 2+)为1-1.2和1-1.6在250-280nm处与叶酸水溶液稍有差异，但不影响UV-B、UV-AII和UV-AI区域内的紫外吸收行为。说明叶酸形成水凝胶后紫外吸收行为并没有明显改变，仍具有紫外吸光的能力。 After folic acid is prepared as a hydrogel, its UV absorption behavior is basically the same as that of folic acid aqueous solution, except that folic acid-zinc ion (F-Zn 2+ ) is 1-1.2 and 1-1.6 at 250-280nm compared with folic acid aqueous solution There is a slight difference, but it does not affect the UV absorption behavior in the UV-B, UV-AII and UV-AI regions. It shows that the ultraviolet absorption behavior of folic acid does not change significantly after the hydrogel is formed, and it still has the ability to absorb ultraviolet light.
实验例9不同防晒组分防晒效果的比较Experimental Example 9 Comparison of sunscreen effects of different sunscreen components
使用对紫外光敏感的试纸(试纸购买自北京博大格林高科技有限公司，型号为日光紫)，在试纸表面涂布防晒组分，夏日中午日光照射(日光强度为0.1W/cm 2)30分钟。 Use test paper sensitive to ultraviolet light (test paper purchased from Beijing Boda Green High-tech Co., Ltd., model is Daylight Violet), coated with sunscreen component on the surface of the test paper, and exposed to sunlight at noon in summer (sunlight intensity is 0.1W/cm 2 ) 30 minute.
照射日光前，试纸显示白色；照射日光后，未涂防晒组分的试纸变为紫色，涂布防晒组分的部位是否变为紫色可以反映防晒成分的防晒性能，即越接近原来的白色表明防晒性能越好。Before exposure to sunlight, the test paper shows white; after exposure to sunlight, the test paper without sunscreen component turns purple. Whether the part where the sunscreen component is applied turns purple can reflect the sunscreen performance of the sunscreen component, that is, the closer to the original white, the sunscreen The better the performance.
实验条件：将阿伏苯宗、BP-3、水杨酸异辛酯、OMC四种防晒组分分别溶解于乙酸乙酯中，并涂布于试纸表面，涂抹面积基本相当，待乙酸乙酯挥发后即可得到涂布防晒组分的试纸。实施例4制备的叶酸-Zn 2+水凝胶直接涂布在试纸表面。按照试纸上防晒组分质量0.02mg,0.1mg,0.2mg,1mg,2mg分组试验。 Experimental conditions: The four sunscreen components of Avobenzone, BP-3, isooctyl salicylate, and OMC were dissolved in ethyl acetate and coated on the surface of the test paper. The smearing area is basically the same. After volatilization, test paper coated with sunscreen components can be obtained. The folic acid-Zn 2+ hydrogel prepared in Example 4 was directly coated on the surface of the test paper. According to the mass of sunscreen components on the test paper, 0.02mg, 0.1mg, 0.2mg, 1mg, 2mg were tested in groups.
结果如图17所示，可以发现：The result is shown in Figure 17, you can find:
(1)在防晒组分剂量为0.02mg和0.1mg时，叶酸-Zn 2+水凝胶可起到防晒的效果，其防晒效果可与阿伏苯宗相比拟，明显高于其他防晒组分。 (1) When the dose of sunscreen component is 0.02mg and 0.1mg, folic acid-Zn 2+ hydrogel can play a sunscreen effect, and its sunscreen effect is comparable to that of avobenzone and is significantly higher than other sunscreen components .
(2)在防晒组分剂量为0.2mg、1mg和2mg时，叶酸-Zn 2+水凝胶的防晒效果与阿伏苯宗、BP-3相似，明显高于其他防晒组分。 (2) When the sunscreen component doses are 0.2mg, 1mg and 2mg, the sunscreen effect of folic acid-Zn 2+ hydrogel is similar to that of Avobenzone and BP-3, and is significantly higher than other sunscreen components.
(3)不论防晒剂使用量是0.02mg、0.1mg、0.2mg、1mg或者2mg，叶酸-Zn 2+水凝胶的防晒效果均明显强于水杨酸异辛酯和OMC。 (3) Regardless of whether the amount of sunscreen is 0.02mg, 0.1mg, 0.2mg, 1mg or 2mg, the sunscreen effect of folic acid-Zn 2+ hydrogel is significantly stronger than that of isooctyl salicylate and OMC.
实验例10叶酸-Zn 2+水凝胶及其与氧化锌复配的防晒效果比较 Experimental example 10 Folic acid-Zn 2+ hydrogel and its sunscreen effect comparison with zinc oxide
紫外光敏感试纸、光照条件和光照时间同实验例9。The ultraviolet light sensitive test paper, light conditions and light time are the same as in Experimental Example 9.
使用实施例4制备的叶酸-Zn 2+水凝胶，向其中分别加入质量分数为0.5％、1％、2％、5％的ZnO粉末，混合均匀，得到的叶酸-Zn 2+水凝胶/ZnO复配防晒剂。 Using the folic acid-Zn 2+ hydrogel prepared in Example 4, 0.5%, 1%, 2%, and 5% of ZnO powder were added to the folic acid-Zn 2+ hydrogel and mixed uniformly to obtain the folic acid-Zn 2+ hydrogel /ZnO compound sunscreen.
配制质量分数分别为0.5％、1％、2％、5％的ZnO水悬浮液作为对照。An aqueous suspension of ZnO with mass fractions of 0.5%, 1%, 2%, and 5% was prepared as a control.
实验条件：分别取100mg的ZnO水悬浮液和叶酸-Zn 2+水凝胶/ZnO复配防晒剂，涂布于试纸上端表面，涂布面积基本为1cm×1cm。 Experimental conditions: Take 100 mg of ZnO aqueous suspension and folic acid-Zn 2+ hydrogel/ZnO compound sunscreen, respectively, and apply them on the upper surface of the test paper. The coating area is basically 1cm×1cm.
实验结果如图18所示，可以发现：0.5％、1％、2％的ZnO涂布后无法起到完全的防晒作用，涂布区域仍有紫色出现；而不同氧化锌质量分数的叶酸-Zn 2+水凝胶/ZnO复配防晒剂则可以起到完全的防晒效果，与5％的ZnO水悬浮液防晒效果相当，甚至防晒效果更佳。 The experimental results are shown in Figure 18. It can be found that 0.5%, 1%, and 2% of ZnO cannot provide complete sun protection after coating, and purple still appears in the coated area; and folic acid-Zn with different zinc oxide mass fractions The 2+ hydrogel/ZnO compound sunscreen can achieve complete sunscreen effect, which is equivalent to or even better with 5% ZnO aqueous suspension.
实验例11叶酸-Zn 2+水凝胶/氧化锌复配与商用防晒剂的防晒效果比较 Experimental Example 11 Comparison of the sunscreen effect of folic acid-Zn 2+ hydrogel/zinc oxide compound and commercial sunscreen
紫外光敏感试纸、光照条件和光照时间同实验例9。The ultraviolet light sensitive test paper, light conditions and light time are the same as in Experimental Example 9.
试验组：实施例4制备的叶酸-Zn 2+水凝胶，向其中加入质量分数5％的ZnO粉末，混合均匀，制得叶酸-Zn 2+水凝胶/5％-ZnO复配防晒剂。 Test group: Folic acid-Zn 2+ hydrogel prepared in Example 4, adding 5% mass fraction of ZnO powder to it, mixing uniformly to prepare folic acid-Zn 2+ hydrogel/5%-ZnO compound sunscreen .
对照组：商用防晒剂使用菲诗小铺(THE FACE SHOP)“活力骄阳水润清透防晒乳”(Vita Sun Block Aqua Sun Lotion)，SPF50+，PA+++。Control group: Commercial sunscreens used THE FACE SHOP "Vita Sun Block Aqua Sun Lotion" (Vita Sun Block Aqua Sun Lotion), SPF50+, PA+++.
实验条件：取商用防晒剂50mg按照2.5mg/cm 2的涂布量均匀涂抹于试纸表面；取叶酸-Zn 2+水凝胶/5％-ZnO复配防晒剂20mg按照1mg/cm 2的涂布量均匀涂抹于试纸表面。涂布成半径约为2.5cm的圆。 Experimental conditions: Take 50mg of commercial sunscreen and apply 2.5mg/cm 2 evenly on the surface of the test paper; take 20mg of folic acid-Zn 2+ hydrogel/5%-ZnO compound sunscreen according to 1mg/cm 2 Spread the cloth evenly on the surface of the test paper. Coated into a circle with a radius of approximately 2.5 cm.
实验结果如图19所示，可以发现：在商用防晒剂的涂布区域中出现了更为明显的紫色区域，可见，使用1mg/cm 2的叶酸-Zn 2+水凝胶/5％-ZnO复配防晒剂与商用防晒剂相比，防晒效果相当甚至更佳。 The experimental results are shown in Figure 19, and it can be found that more obvious purple areas appear in the coating area of commercial sunscreens. It can be seen that 1mg/cm 2 of folic acid-Zn 2+ hydrogel/5%-ZnO is used Compared with commercial sunscreens, compound sunscreens have the same or better sunscreen effect.
以上结合具体实施方式和范例性实例对本发明进行了详细说明，不过这些说明并不能理解为对本发明的限制。本领域技术人员理解，在不偏离本发明精神和范围的情况下，可以对本发明技术方案及其实施方式进行多种等价替换、修饰或改进，这些均落入本发明的范围内。本发明的保护范围以所附权利要求为准。The present invention has been described in detail above with reference to specific embodiments and exemplary examples, but these descriptions should not be understood as limiting the present invention. Those skilled in the art understand that without departing from the spirit and scope of the present invention, various equivalent substitutions, modifications or improvements can be made to the technical solutions and embodiments of the present invention, and these all fall within the scope of the present invention. The protection scope of the present invention is subject to the appended claims.

Claims (10)

  1. 叶酸-金属离子水凝胶，其特征在于，该水凝胶具有网状结构，包括配位连接的金属离子和叶酸分子；Folic acid-metal ion hydrogel, characterized in that the hydrogel has a network structure, including coordinated metal ions and folic acid molecules;
    叶酸的喋呤头基通过氢键形成四联体，四联体之间π-π堆积形成纤维，金属离子和叶酸的羧酸配位，使纤维交联并相互缠绕形成网状结构。The pterin head group of folic acid forms a quadruplex through hydrogen bonding, and the π-π stacks between the tetrads to form fibers, and the metal ions are coordinated with the carboxylic acid of folic acid, so that the fibers are cross-linked and intertwined to form a network structure.
  2. 根据权利要求1所述的叶酸-金属离子水凝胶，其特征在于，在所述叶酸-金属离子水凝胶的XRD图谱中，在约8.5°和约26.5°处存在特征峰；和/或，The folic acid-metal ion hydrogel of claim 1, wherein, in the XRD pattern of the folic acid-metal ion hydrogel, there are characteristic peaks at about 8.5° and about 26.5°; and/or,
    所述水凝胶在圆二色谱的谱图中存在叶酸的手性堆积峰；特别地，在300nm附近和260nm附近显示手性信号。The hydrogel has a chiral stacking peak of folic acid in the spectrum of the circular dichroism; in particular, it shows chiral signals near 300 nm and 260 nm.
  3. 叶酸-金属离子水凝胶，其特征在于，该水凝胶由包括以下步骤制备：The folic acid-metal ion hydrogel is characterized in that the hydrogel is prepared by the following steps:
    (1)将叶酸溶解于水或水性缓冲溶液中，配制叶酸水溶液；(1) Dissolve folic acid in water or an aqueous buffer solution to prepare an aqueous folic acid solution;
    (2)将金属盐溶解于水或水性缓冲溶液中，配制金属离子水溶液；(2) Dissolve the metal salt in water or an aqueous buffer solution to prepare an aqueous solution of metal ions;
    (3)将叶酸水溶液和金属离子水溶液室温下均匀混合，静置，即得叶酸-金属离子水凝胶；(3) The folic acid aqueous solution and the metal ion aqueous solution are uniformly mixed at room temperature and left to stand to obtain folic acid-metal ion hydrogel;
    所述叶酸-金属离子水凝胶为溶液、凝胶或混悬液状态。The folic acid-metal ion hydrogel is in the state of solution, gel or suspension.
  4. 根据权利要求3所述的叶酸-金属离子水凝胶，其特征在于，所述金属盐为盐酸盐、硝酸盐或硫酸盐；金属盐中金属离子选自锌、银、镁和钙离子中的一种或多种；和/或，The folic acid-metal ion hydrogel according to claim 3, wherein the metal salt is hydrochloride, nitrate or sulfate; the metal ions in the metal salt are selected from zinc, silver, magnesium and calcium ions One or more of; and/or,
    步骤(1)中，水或水性缓冲溶液溶解叶酸时，调节使水或水性缓冲溶液的pH值为7.0～9.5；和/或，In step (1), when water or aqueous buffer solution dissolves folic acid, adjust the pH of the water or aqueous buffer solution to a value of 7.0 to 9.5; and/or,
    步骤(1)和(2)中，水性缓冲溶液选自乙酸-乙酸钠、硼酸-硼砂、N-(2-羟乙基)哌嗪-N′-2-乙烷磺酸、3-吗啉丙磺酸、2-(N-吗啡啉)乙磺酸、三羟甲基甲胺-盐酸、三羟甲基甲胺-硼酸、三羟甲基甲胺-乙酸中的一种或多种；和/或，In steps (1) and (2), the aqueous buffer solution is selected from acetic acid-sodium acetate, boric acid-borax, N-(2-hydroxyethyl)piperazine-N′-2-ethanesulfonic acid, 3-morpholine One or more of propanesulfonic acid, 2-(N-morpholine)ethanesulfonic acid, trimethylolmethylamine-hydrochloric acid, trimethylolmethylamine-boronic acid, and trimethylolmethylamine-acetic acid; and / or,
    步骤(3)中，叶酸水溶液和金属离子水溶液混合后，叶酸的最终浓度为0.5mM-200mM，优选为5mM-100mM；In step (3), after the aqueous folic acid solution and the metal ion aqueous solution are mixed, the final concentration of folic acid is 0.5 mM-200 mM, preferably 5 mM-100 mM;
    金属离子的最终浓度为0.1mM-500mM，优选为5mM-400mM；和/或，The final concentration of metal ions is 0.1mM-500mM, preferably 5mM-400mM; and/or,
    更优选步骤(3)中，叶酸水溶液和金属离子水溶液混合后，叶酸和金属离子的摩尔浓度比为1∶0.1-10，优选为1∶1-5。More preferably, in step (3), after the folic acid aqueous solution and the metal ion aqueous solution are mixed, the molar concentration ratio of folic acid and metal ion is 1:0.1-10, preferably 1:1-5.
  5. 根据权利要求3所述的叶酸-金属离子水凝胶，其特征在于，金属离子为锌离子时，制得叶酸-锌离子水凝胶；The folic acid-metal ion hydrogel according to claim 3, wherein when the metal ion is zinc ion, folic acid-zinc ion hydrogel is prepared;
    将叶酸水溶液和锌离子水溶液混合后，随着叶酸浓度的提高，叶酸-锌离子水凝胶形成凝胶态时的叶酸/锌离子摩尔比下限逐渐降低，上限逐渐提高，凝胶区域扩大；After mixing the folic acid aqueous solution and the zinc ion aqueous solution, as the concentration of folic acid increases, the lower limit of the folate/zinc ion molar ratio when the folate-zinc ion hydrogel forms a gel state gradually decreases, the upper limit gradually increases, and the gel area expands;
    叶酸浓度不变时，叶酸/锌离子摩尔比逐渐增大，叶酸-锌离子水凝胶由溶液状态变为凝胶状态，再变成混悬液状态。When the concentration of folic acid remains unchanged, the molar ratio of folic acid/zinc ion gradually increases, and the folic acid-zinc ion hydrogel changes from a solution state to a gel state, and then to a suspension state.
  6. 防晒剂，其含有权利要求1-5之一所述叶酸-金属离子水凝胶，所述防晒剂中还包括氧化锌；A sunscreen agent, which contains the folic acid-metal ion hydrogel according to any one of claims 1-5, and the sunscreen agent further includes zinc oxide;
    所述氧化锌在防晒剂中的质量分数为0.1％-10％，优选为0.5％-5％；The mass fraction of the zinc oxide in the sunscreen is 0.1%-10%, preferably 0.5%-5%;
    所述防晒剂为膏剂、霜剂或凝胶剂。The sunscreen is ointment, cream or gel.
  7. 权利要求1或2所述叶酸-金属离子水凝胶的制备方法，包括以下步骤：The preparation method of folic acid-metal ion hydrogel according to claim 1 or 2, comprising the following steps:
    (1)配制叶酸水溶液；(1) Prepare an aqueous solution of folic acid;
    (2)配制金属离子水溶液；(2) Preparation of metal ion aqueous solution;
    (3)将叶酸水溶液和金属离子水溶液室温下均匀混合，静置，即得水凝胶。(3) The folic acid aqueous solution and the metal ion aqueous solution are uniformly mixed at room temperature and left to stand to obtain a hydrogel.
  8. 根据权利要求7所述的制备方法，其特征在于，在步骤(1)中，将叶酸溶解于水性溶液中配制叶酸水溶液；所述水性溶液包括水或水性缓冲溶液；和/或，The preparation method according to claim 7, wherein in step (1), folic acid is dissolved in an aqueous solution to prepare an aqueous folic acid solution; the aqueous solution includes water or an aqueous buffer solution; and/or,
    在步骤(2)中，所述金属离子选自一价或二价态时化学性质稳定的金属离子；优选地，所述金属离子选自钠、钾、锂、铷、铯、锌(Zn 2+)，铜(Cu 2+)，镍(Ni 2+)，钴(Co 2+)，锰(Mn 2+)，铬(Cr 2+)，钼(Mo 2+)，银(Ag +)，镉(Cd 2+)，镁(Mg 2+)，钙(Ca 2+)，铅(Pb 2+)、钡、金属中的一种或多种；和/或， In step (2), the metal ion is selected from metal ions that are chemically stable in a monovalent or divalent state; preferably, the metal ion is selected from sodium, potassium, lithium, rubidium, cesium, zinc (Zn 2 + ), copper (Cu 2+ ), nickel (Ni 2+ ), cobalt (Co 2+ ), manganese (Mn 2+ ), chromium (Cr 2+ ), molybdenum (Mo 2+ ), silver (Ag + ) , Cadmium (Cd 2+ ), magnesium (Mg 2+ ), calcium (Ca 2+ ), lead (Pb 2+ ), barium, one or more of metals; and/or,
    在步骤(3)中，混合后，所述叶酸在混合体系中的最终浓度为0.5mM-200mM，优选为5mM-100mM；In step (3), after mixing, the final concentration of the folic acid in the mixing system is 0.5mM-200mM, preferably 5mM-100mM;
    所述金属离子在混合体系中的最终浓度为0.5mM-500mM，优选为5mM-400mM。The final concentration of the metal ion in the mixed system is 0.5mM-500mM, preferably 5mM-400mM.
  9. 权利要求1-5之一所述叶酸-金属离子水凝胶或权利要求7或8所述方法制备的水凝胶在生物材料中的用途；优选在细胞三维培养中的用途或生物三维打印中的用途。Use of the folate-metal ion hydrogel according to any one of claims 1 to 5 or the hydrogel prepared by the method according to claim 7 or 8 in biological materials; preferably in three-dimensional cell culture or biological three-dimensional printing the use of.
  10. 权利要求1-5之一所述叶酸-金属离子水凝胶和权利要求6所述防晒剂在预防或防止紫外吸收中的用途；优选用作防晒剂的用途；和/或，Use of the folic acid-metal ion hydrogel of any one of claims 1 to 5 and the sunscreen of claim 6 for preventing or preventing ultraviolet absorption; preferably for use as a sunscreen; and/or,
    叶酸-金属离子组合物在预防或防止紫外吸收中的用途，其特征在于，所述叶酸-金属离子组合物包括叶酸，和锌、银、镁和钙离子中的一种或多种；The use of a folic acid-metal ion composition for preventing or preventing ultraviolet absorption, characterized in that the folic acid-metal ion composition includes folic acid, and one or more of zinc, silver, magnesium and calcium ions;
    优选所述叶酸-金属离子组合物为叶酸-金属离子水凝胶。Preferably, the folic acid-metal ion composition is folic acid-metal ion hydrogel.
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