Application of zero-valent iodine in the detection of hyaluronic acid

Abstract

The present invention discloses an application of zero-valent iodine in the detection of hyaluronic acid, wherein zero-valent iodine is mixed with a sample to be detected in a solvent, and the presence of hyaluronic acid in the sample to be detected is determined by the color change of the mixture. The present invention can realize rapid detection of HA, and is convenient to operate, time-saving, and low-cost. The present invention can be effectively suitable for the detection of HA in products such as cosmetics, food, paper products, non-woven products, and fabrics (which are not easy to use traditional HPLC detection methods), and has a good application prospect.

Description

Technical Field

[0001] The invention relates to application of zero-valent iodine in hyaluronic acid detection, belonging to the technical field of chemical detection. Background Art

[0002] Hyaluronic acid (HA), also known as hyaluronic acid, is a high molecular weight straight-chain mucopolysaccharide composed of (1-3)-2-acetylamino-2-deoxy-D-glucose (1-4)-DD-glucuronic acid disaccharide repeating units. Hyaluronic acid has good moisturizing properties and has been added to various skin care products, daily necessities, and foods in recent years, such as essences, facial masks, paper towels, wet wipes, non-woven products, fabric products, beverages, and the like. The HA detection method recorded in the European Pharmacopoeia is currently the most commonly used method for HA detection. It is necessary to first degrade HA into glucuronic acid, and then use the color reaction of glucuronic acid and carbazole to calculate the content of glucuronic acid by photometry, and then convert the content of HA. The reaction process of this method is complicated and time-consuming, and a strong acid is also needed, which also pollutes the environment to a certain extent.

[0003] HA in products can also be detected by high-performance liquid chromatography, but this method generally requires a series of "extraction-enzymatic hydrolysis-HPLC" operations. For example, patent CN109298112B provides a method for determining the content of hyaluronic acid, which is suitable for the determination of hyaluronic acid content in health foods, medicines, medical devices, cosmetics, hair care products, oral care products, and paper products. It is necessary to first use hyaluronidase to enzymatically hydrolyze the sample containing hyaluronic acid and then use high-performance liquid chromatography to determine the HA content. Not only is it time-consuming, but it is also limited by the equipment model and cannot quickly detect HA in the product.

[0004] Currently, no method has been found in the prior art that can quickly detect HA in products. Summary of the invention

[0005] In view of the shortcomings of the prior art, the inventors have surprisingly found, after a large number of experiments and studies, that hyaluronic acid and zero-valent iodine can change color after mixing. The color of zero-valent iodine is yellow. After mixing with hyaluronic acid, the yellow color disappears and the mixed solution becomes colorless. The inventors further studied other polyhydroxy polysaccharides with similar structures to hyaluronic acid, such as chitosan and sodium alginate, and found that under the same conditions, other polyhydroxy polysaccharides did not change color when mixed with zero-valent iodine. Therefore, the color reaction between zero-valent iodine and hyaluronic acid has certain specificity, and zero-valent iodine can be used to indicate the presence of hyaluronic acid.

[0006] The specific technical solutions of this application are as follows:

[0007] The present invention provides an application of zero-valent iodine in detecting hyaluronic acid.

[0008] In some embodiments, zerovalent iodine, a sample to be tested, and a solvent are mixed, and the presence of hyaluronic acid is determined by the change in color of the mixture.

[0009] The zero-valent iodine includes one or more of elemental iodine and a compound of elemental iodine and a polymer.

[0010] The compound of iodine element and polymer includes povidone iodine.

[0011] The present invention also provides an application of a composition containing zerovalent iodine in detecting hyaluronic acid.

[0012] The composition comprises zero-valent iodine and a solvent, wherein the zero-valent iodine comprises one or more of simple iodine and a complex of simple iodine and a polymer, and the solvent comprises water.

[0013] The present invention also provides a method for detecting hyaluronic acid, which comprises mixing a sample to be detected, zero-valent iodine and a solvent, and observing the color change of the mixture.

[0014] The samples to be tested include cosmetics, foods, health products, paper products, non-woven products, fabric products, and / or their extracts.

[0015] In some embodiments, zero-valent iodine is mixed with a sample to be tested in a solvent, and the presence of hyaluronic acid is determined by the color change of the mixture. The sample to be tested is preferably a product expected to contain hyaluronic acid.

[0016] Furthermore, since zero-valent iodine is yellow, when the color of the mixture remains unchanged from the initial color, it indicates that no hyaluronic acid is present, and when the initial color of the mixture changes, it indicates that hyaluronic acid is present.

[0017] In some embodiments, when the mixture also contains other colored ingredients, such as dyes, the mixture containing hyaluronic acid may have multiple color types. The dyes that can be added are water-soluble dyes such as CI 45430, CI 16255, CI15510, CI 45410, CI 16035, CI42090, etc.

[0018] The present invention is based on the phenomenon that iodine and hyaluronic acid substances can react to produce color change, and the presence of hyaluronic acid substances can be judged by naked eyes, thereby realizing rapid qualitative detection of hyaluronic acid substances.

[0019] The hyaluronic acid mentioned in the present invention refers to hyaluronic acid substances, which include hyaluronic acid, hyaluronate, cross-linked hyaluronic acid, cross-linked hyaluronate, hyaluronic acid derivatives, cross-linked hyaluronic acid derivatives and other hyaluronic acid substances. Among them, hyaluronate includes sodium hyaluronate, potassium hyaluronate, zinc hyaluronate, magnesium hyaluronate, calcium hyaluronate, cobalt hyaluronate, gold hyaluronate and the like. Hyaluronic acid derivatives include acetylated hyaluronic acid, thiolated hyaluronic acid and the like, and cross-linked hyaluronic acid derivatives include acetylated cross-linked hyaluronic acid, thiolated cross-linked hyaluronic acid and the like.

[0020] The zero-valent iodine includes iodine element and / or a compound of iodine element and a polymer. The compound of iodine element and polymer is not particularly limited, and may be, for example, povidone iodine, etc. The color of zero-valent iodine is yellow. When the sample solution to be tested is mixed with zero-valent iodine, the solution is yellow. When the sample contains hyaluronic acid, iodine reacts with the hyaluronic acid, the color of zero-valent iodine becomes lighter or disappears, and the color of the mixed solution becomes lighter or directly becomes colorless.

[0021] In some embodiments, the sample to be tested is contacted and mixed with zerovalent iodine in the form of a solution. When the sample to be tested can be dissolved in a solvent, the sample to be tested, zerovalent iodine and the solvent are mixed. The order of mixing can be selected at will. For example, the sample to be tested and the solvent can be prepared into a solution, and the zerovalent iodine and the solvent can be prepared into a solution, and then the two solutions are mixed; or, the sample to be tested and the zerovalent iodine are directly dissolved in the solvent at the same time and mixed evenly. When the sample to be tested cannot be completely dissolved directly in the solvent, the sample to be tested needs to be extracted with a solvent in advance, and then the extract is mixed with zerovalent iodine. The solvent is, for example, water.

[0022] Once zero-valent iodine and hyaluronic acid come into contact, they will react. The more iodine is added or the more hyaluronic acid is contained, the faster the reaction and the faster the color change. However, it is not easy to add too much iodine. If it is added too much, there will be excess iodine in the mixed system, and it is difficult to observe the color change. The mass ratio of hyaluronic acid to elemental iodine may be (1000-0.1):1, preferably (100-1):1, for example, 1000:1, 900:1, 800:1, 700:1, 600:1, 500:1, 400:1, 300:1, 200:1, 100:1, 90:1, 80:1, 70:1, 60:1, 50:1, 40:1, 30:1, 20:1, 10:1, 9:1, 8:1, 7:1, 6:1, 5:1, 4:1, 3:1, 2:1, 1:1, 0.9:1, 0.8:1, 0.7:1, 0.6:1, 0.5:1, 0.4:1, 0.3:1, 0.2:1, 0.1:1. The content of zero-valent iodine can be selected in a conventional amount. In some embodiments, the content of zero-valent iodine is 0.0001% to 0.1%, preferably 0.001% to 0.01%, for example, it can be 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%; the content of HA can be HA in cosmetics, food, health products, paper products, non-woven products, textiles The conventional addition amount in products such as pharmaceuticals, in some embodiments, the content of HA is 0.0001% to 5%, preferably 0.001% to 1.0%, for example, it can be 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.001%, 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.007%, 0.008%, 0.009%, 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%.

[0023] The present invention also provides an application of zero-valent iodine in the detection of hyaluronic acid, which can be used for the qualitative detection of HA in products, such as cosmetics, foods, health products, paper products, non-woven products, fabrics, etc., and / or extracts thereof. When hyaluronic acid is mixed with zero-valent iodine, a color change visible to the naked eye may occur, which takes a short time and can achieve rapid qualitative detection of HA.

[0024] In some embodiments, the present invention can be used to determine whether a product contains hyaluronic acid, and can also be used to analyze the HA content between different products. For example, the same type of products produced by different manufacturers containing HA can be tested for HA using this qualitative method, and the speed of color change can be used to evaluate which product has a higher hyaluronic acid content.

[0025] In some embodiments, the present invention can also be used for quality control in the production process. For example, in the production process of beverages containing HA, the qualitative method can be used to quickly detect whether HA is contained, which is used to control product quality.

[0026] In some embodiments, the present invention can also be used to quickly distinguish genuine and fake goods in the market. The method is simple to operate, has intuitive phenomena, is short in time consumption, is highly efficient, does not require the assistance of other equipment, is low in cost, and has high application value.

[0027] In some embodiments, since the color change phenomenon of the present invention is intuitive, it can also be used for experimental demonstration, allowing consumers to intuitively see that HA ingredients are added to the product.

[0028] The present invention has the following beneficial effects:

[0029] 1. The present invention provides an application of zero-valent iodine in the detection of hyaluronic acid. By mixing zero-valent iodine with hyaluronic acid, a color change visible to the naked eye can occur, which can achieve rapid detection of HA, with easy operation, short time consumption and low cost.

[0030] 2. The present invention is effectively suitable for the detection of HA in cosmetics, foods, health products, paper products, non-woven products, fabric products, etc. that are expected to contain HA (which are not easy to detect using traditional HPLC methods). Before quantitatively detecting the HA content in the product, it can be used for the preliminary screening of whether the product contains HA, and has a good application prospect.

[0031] 3. The present invention can be applied to a variety of different scenario requirements and realize rapid qualitative detection of HA. It has the advantages of simple operation, intuitive phenomenon, high efficiency, and no need for other equipment assistance. DETAILED DESCRIPTION

[0032] The present invention is further described below by means of specific examples. The following description is only exemplary and does not limit the content thereof.

[0033] In the following examples, all reagents used are commercially available products, and some of the reagents are shown in Table 1 below:

[0034]

[0035] Example 1

[0036] Weigh 0.002 g of sodium hyaluronate (30 wDa) and dissolve it in purified water, and dilute to 100 ml to prepare a hyaluronic acid solution with a mass percentage of 0.002%;

[0037] Weigh 0.002 g of povidone iodine and dissolve it in purified water, dilute to 100 ml, and prepare a povidone iodine solution with a mass percentage of 0.002%;

[0038] The above hyaluronic acid solution and povidone-iodine solution were mixed in equal volumes, and the mass percentages of sodium hyaluronate and povidone-iodine in the mixture after mixing were as shown in Table 2.

[0039] Example 2-14

[0040] According to the steps of Example 1, sodium hyaluronate, cross-linked sodium hyaluronate or acetylated sodium hyaluronate are dissolved in purified water to prepare a hyaluronic acid aqueous solution. Zero-valent iodine is also dissolved in purified water to prepare a zero-valent iodine aqueous solution.

[0041] The hyaluronic acid aqueous solution and the zero-valent iodine aqueous solution were mixed in a certain volume ratio to obtain a mixture solution with different hyaluronic acid and zero-valent iodine concentrations as shown in Table 2.

[0042]

[0043] Comparative Example 1

[0044] Sodium alginate solution: weigh 0.002 g of sodium alginate and dissolve it in purified water, dilute to 100 ml, and prepare a sodium alginate solution with a mass percentage of 0.002%;

[0045] Povidone-iodine solution: weigh 0.002 g of povidone-iodine and dissolve it in purified water, dilute to 100 ml, and prepare a povidone-iodine solution with a mass percentage of 0.002%;

[0046] The sodium alginate solution and the povidone iodine solution are mixed in equal volumes, and the mass percentages of sodium alginate and povidone iodine in the mixture are both 0.001%.

[0047] Comparative Example 2

[0048] Chitosan solution: weigh 0.002 g of chitosan and dissolve it in purified water, dilute to 100 ml, and prepare a chitosan solution with a mass percentage of 0.002%;

[0049] Povidone-iodine solution: weigh 0.002 g of povidone-iodine and dissolve it in purified water, dilute to 100 ml, and prepare a povidone-iodine solution with a mass percentage of 0.002%;

[0050] The chitosan solution and the povidone-iodine solution are mixed in equal volumes, and the mass percentages of chitosan and povidone-iodine in the mixture are both 0.001%.

[0051] Color change experiment

[0052] The time after the hyaluronic acid aqueous solution and the zero-valent iodine aqueous solution are mixed is defined as the initial time, and the solution color is observed to see if there is a change. If there is a change, the time required for the solution color to change to colorless from the initial time is recorded, which is the color change time. The color change time of Comparative Example 1 is the time required for the solution color to change to colorless after the sodium alginate solution and the povidone-iodine solution are mixed. The color change time of Comparative Example 2 is the time required for the solution color to change to colorless after the chitosan solution and the povidone-iodine solution are mixed.

[0053] The color change of the mixture solution of each embodiment and comparative example is shown in Table 3 below:

[0054]

[0055] From the above experiments, it can be seen that sodium hyaluronate, cross-linked sodium hyaluronate and sodium hyaluronate derivatives of different concentrations and molecular weights all change color after mixing with zero-valent iodine, while sodium alginate and chitosan, which have a similar structure to hyaluronic acid, cannot undergo a color reaction with zero-valent iodine under the same conditions. This shows that the color reaction between zero-valent iodine and HA has a certain specificity, and zero-valent iodine can specifically indicate the presence of HA.

Claims

1. An application of zero-valent iodine in detecting hyaluronic acid, characterized in that: Zero-valent iodine, a sample to be tested and a solvent are mixed, and the presence of hyaluronic acid is determined by the change in the color of the mixture, wherein the solvent includes water.

2. The use according to claim 1, characterized in that: The zero-valent iodine includes one or more of elemental iodine and a compound of elemental iodine and a polymer.

3. The use according to claim 2, characterized in that: The compound of iodine element and polymer includes povidone iodine.

4. The use according to any one of claims 1 to 3, characterized in that: The samples to be tested include cosmetics, foods, health products, paper products, non-woven products, fabric products, and / or extracts thereof.

5. Use of a composition comprising zerovalent iodine in detecting hyaluronic acid, characterized in that: The composition includes zerovalent iodine and a solvent, and the solvent includes water.

6. The use according to claim 5, characterized in that: The zero-valent iodine includes one or more of elemental iodine and a compound of elemental iodine and a polymer.

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