Modified hydroxytyrosol, preparation method and application

Abstract

The invention discloses modified hydroxytyrosol as well as a preparation method and application thereof. The provided preparation method of the modified hydroxytyrosol comprises the following steps: in the presence of an acidic solvent, mixing hydroxytyrosol, protease and collagen for reaction, mixing a reaction product with an extraction agent, performing centrifugal layering, and collecting upper-layer components, thereby obtaining the modified hydroxytyrosol, the protease comprises one or two of neutral protease and trypsin. Under the enzymolysis action of protease, the collagen can be decomposed into amino acid monomers such as glycine, proline and hydroxyproline, and the amino acid monomers can be subjected to graft modification with hydroxytyrosol; meanwhile, collagen fragments which are not subjected to enzymolysis also exist, and are subjected to graft modification with hydroxytyrosol in the form of collagen characteristic fragments. The oxidation resistance and the stability of the obtained modified hydroxytyrosol are obviously improved compared with those of unmodified hydroxytyrosol.

Description

Technical Field

[0001] This application belongs to the field of cosmetic or food preparation technology, and relates to modified hydroxytyrosol, its preparation method and application. Background Technology

[0002] Hydroxytyrosol is a polyphenol found in olive oil. While it possesses free radical scavenging capabilities, its antioxidant properties are currently considered poor. Furthermore, it has been found to be unstable and prone to discoloration at high temperatures, limiting its wider application. Summary of the Invention

[0003] Therefore, it is necessary to provide a modified hydroxytyrosol, its preparation method, and its application to improve the antioxidant properties and stability of hydroxytyrosol.

[0004] In some embodiments, a method for preparing modified hydroxytyrosol for cosmetics or food is provided, comprising the following steps: mixing hydroxytyrosol, protease and collagen in the presence of an acidic solvent to react, mixing the reaction product with an extractant, centrifuging to separate the layers, collecting the upper layer components, and preparing the modified hydroxytyrosol.

[0005] The protease includes one or both of neutral proteases and trypsin.

[0006] In some embodiments, in the provided method for preparing modified hydroxytyrosol for cosmetics or food, the mass ratio of hydroxytyrosol to collagen is 1:(1~2).

[0007] In some embodiments, in the provided method for preparing modified hydroxytyrosol for cosmetics or food, the mass ratio of hydroxytyrosol to the protease is 1:(1~2).

[0008] In some embodiments, in the provided method for preparing modified hydroxytyrosol for cosmetics or food, the pH value of the solvent is 6 to 6.8.

[0009] In some embodiments, in the provided method for preparing modified hydroxytyrosol for cosmetics or food, the reaction temperature is 40°C to 50°C, and the reaction time is 4h to 8h.

[0010] In some embodiments, the provided method for preparing modified hydroxytyrosol for cosmetics or food satisfies one or more of the following conditions:

[0011] (1) The extractant includes ethyl acetate;

[0012] (2) The mass-volume ratio of the hydroxytyrosol to the extractant is 1 g: (2~4) mL.

[0013] In some embodiments, the provided method for preparing modified hydroxytyrosol for cosmetics or food satisfies one or more of the following conditions:

[0014] (1) The solvent is water;

[0015] (2) The mass-to-volume ratio of the hydroxytyrosol to the solvent is 1 g: (8~15) mL;

[0016] (3) In the centrifugation layering step, centrifuge at 5000rpm~8000rpm for 5min~8min.

[0017] In some embodiments, a modified hydroxytyrosol is provided, which is prepared by the aforementioned preparation method.

[0018] In some embodiments, the use of the modified hydroxytyrosol is provided in the preparation of products with free radical scavenging capabilities;

[0019] The products mentioned include cosmetics or food.

[0020] In some embodiments, a cosmetic or food product is provided containing the modified hydroxytyrosol.

[0021] In the aforementioned method for preparing modified hydroxytyrosol, collagen is decomposed into amino acid monomers such as glycine, proline, and hydroxyproline under the enzymatic hydrolysis of proteases. These amino acid monomers are then grafted onto hydroxytyrosol for modification. Simultaneously, undigested collagen fragments are also grafted onto hydroxytyrosol as characteristic collagen fragments. The resulting modified hydroxytyrosol exhibits significantly improved antioxidant properties and stability compared to unmodified hydroxytyrosol. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments and examples of this application, and to more completely understand this application and its beneficial effects, the accompanying drawings used in the description of the embodiments or examples will be briefly introduced below. Obviously, the drawings described below are merely some embodiments of this application. Those skilled in the art can obtain other drawings based on these drawings without any creative effort.

[0023] Figure 1 Infrared spectrum of unmodified hydroxytyrosol;

[0024] Figure 2 The infrared spectrum of the modified hydroxytyrosol in Example 1;

[0025] Figure 3 The mass spectrum of the modified hydroxytyrosol in Example 1;

[0026] Figure 4 The image shows the stability test results of modified hydroxytyrosol, where, from left to right, unmodified hydroxytyrosol and modified hydroxytyrosol are shown respectively. Detailed Implementation

[0027] To facilitate understanding of the present invention, a more complete description will be given below with reference to the accompanying drawings. Preferred embodiments of the invention are shown in the drawings. However, the invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the invention.

[0028] It should be noted that when a component is said to be "fixed to" another component, it can be directly attached to the other component or there may be an intervening component. When a component is said to be "connected to" another component, it can be directly connected to the other component or there may be an intervening component.

[0029] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0030] Unless otherwise stated or in case of contradiction, the terms or phrases used herein shall have the following meanings:

[0031] The terms "and / or," "or / and," and "and / or" as used in this application encompass any one of two or more related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected using at least two conjunctions selected from "and / or," "or / and," and "and / or," it should be understood that in this application, the technical solution undoubtedly includes solutions connected by "logical AND," and also undoubtedly includes solutions connected by "logical OR." For example, "A and / or B" includes three parallel solutions: A, B, and "a combination of A and B."

[0032] In this application, the terms "multiple", "various", "multiple times", "multi-dimensional", etc., unless otherwise specified, refer to a quantity greater than or equal to 2. For example, "one or more" means one or more than or equal to two.

[0033] The terms “combinations thereof,” “any combination thereof,” and “any combination thereof” as used in this application include all suitable combinations of any two or more of the listed items.

[0034] In this application, the term "suitable" as used in "suitable combination", "suitable method", "any suitable method", etc., refers to the ability to implement the technical solution of this application, solve the technical problem of this application, and achieve the expected technical effect of this application.

[0035] In this application, terms such as "preferred," "better," "more suitable," and "ideal" are merely used to describe implementation methods or embodiments that achieve better results, and should be understood not to limit the scope of protection of this application.

[0036] In this application, terms such as "further," "even more," and "particularly" are used for descriptive purposes and to indicate differences in content, but should not be construed as limiting the scope of protection of this application.

[0037] In this application, "optionally," "optionally," and "optional" mean that something is optional, that is, it means that it is selected from either "with" or "without." If there are multiple "optional" entries in a technical solution, unless otherwise specified, and there are no contradictions or mutual constraints, each "optional" entry shall be independent.

[0038] In this invention, the terms "first aspect," "second aspect," "third aspect," and "fourth aspect," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance or quantity, nor should they be construed as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first," "second," "third," and "fourth," etc., serve only as a non-exhaustive enumeration and should be understood not to constitute a closed limitation on quantity.

[0039] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.

[0040] In this application, numerical intervals (i.e., numerical ranges) are involved. Unless otherwise specified, the selected numerical distributions within the aforementioned numerical intervals are considered continuous and include the two endpoints (i.e., the minimum and maximum values) of the numerical range, as well as every value between these two endpoints. Unless otherwise specified, when a numerical interval refers only to integers within that interval, it includes the two endpoint integers of the numerical range, as well as every integer between the two endpoints. In this document, this is equivalent to directly listing every integer. For example, if t is an integer selected from 1 to 10, it means that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. Furthermore, when multiple ranges are provided to describe features or characteristics, these ranges can be merged. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges to which they are included.

[0041] Unless otherwise specified, the temperature parameters in this application are permitted to be either constant-temperature treatment or variations within a certain temperature range. It should be understood that the constant-temperature treatment allows temperature fluctuations within the precision range of the instrument control, such as ±5℃, ±4℃, ±3℃, ±2℃, or ±1℃.

[0042] In this application, % (w / w) and wt% both represent weight percentage, % (v / v) refers to volume percentage, and % (w / v) refers to mass-volume percentage.

[0043] In this application, "room temperature" generally refers to 5℃~30℃, and more preferably 25±5℃.

[0044] In some embodiments, a method for preparing modified hydroxytyrosol for use in cosmetics or food is provided, comprising the following steps: mixing hydroxytyrosol, protease and collagen in the presence of an acidic solvent to react, mixing the reaction product with an extractant, centrifuging to separate the layers, collecting the upper layer components, and preparing modified hydroxytyrosol.

[0045] Proteases include one or both of neutral proteases and trypsin.

[0046] In some embodiments, in the provided method for preparing modified hydroxytyrosol for cosmetics or food, the mass ratio of hydroxytyrosol to collagen is 1:(1~2), for example, the mass ratio of hydroxytyrosol to collagen is 1:1, 1:1.5, 1:2, etc., or it can be a range of any two of the aforementioned ratios.

[0047] In some embodiments, in the provided method for preparing modified hydroxytyrosol for cosmetics or food, the mass ratio of hydroxytyrosol to protease is 1:(1~2), for example, the mass ratio of hydroxytyrosol to protease is 1:1, 1:1.5, 1:2, etc., or it can be a range of any two of the aforementioned ratios.

[0048] In some embodiments, in the provided method for preparing modified hydroxytyrosol for cosmetics or food, the pH value of the solvent is 6 to 6.8. For example, the pH value of the solvent is 6, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, etc., or it can be a range composed of any two of the aforementioned values.

[0049] In some embodiments, the method for preparing modified hydroxytyrosol for cosmetics or food uses a reaction temperature of 40°C to 50°C and a reaction time of 4 hours to 8 hours. For example, the reaction temperature can be 40°C, 45°C, 50°C, or any combination of the aforementioned values. The reaction time can be 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, or any combination of the aforementioned values.

[0050] In some embodiments, the extraction agent in the provided method for preparing modified hydroxytyrosol for cosmetics or food includes ethyl acetate.

[0051] In some embodiments, in the provided method for preparing modified hydroxytyrosol for cosmetics or food, the mass-volume ratio of hydroxytyrosol to extractant is 1g:(2~4)mL. For example, the mass-volume ratio of hydroxytyrosol to extractant can be 1g:2mL, 1g:3mL, 1g:4mL, etc., or it can be a range of any two of the aforementioned ratios.

[0052] In some embodiments, the solvent provided in the method for preparing modified hydroxytyrosol for cosmetics or food is water.

[0053] In some embodiments, in the provided method for preparing modified hydroxytyrosol for cosmetics or food, the mass-volume ratio of hydroxytyrosol to solvent is 1g:(8~15)mL. For example, the mass-volume ratio of hydroxytyrosol to solvent can be 1g:8mL, 1g:9mL, 1g:10mL, 1g:11mL, 1g:12mL, 1g:13mL, 1g:14mL, 1g:15mL, etc., or it can be a range of any two of the aforementioned ratios.

[0054] In some embodiments, in the method for preparing modified hydroxytyrosol for cosmetics or food, the centrifugation separation step involves centrifugation at 5000 rpm to 8000 rpm for 5 min to 8 min.

[0055] In some embodiments, a modified hydroxytyrosol is provided, which is prepared by the aforementioned preparation method.

[0056] In some embodiments, the use of modified hydroxytyrosol in the preparation of products with free radical scavenging capabilities is provided;

[0057] Products include cosmetics or food.

[0058] In some embodiments, a cosmetic or food product is provided containing modified hydroxytyrosol.

[0059] The following are specific embodiments. They are intended to provide a more detailed description of this application to help those skilled in the art and researchers better understand it. The technical conditions described do not constitute any limitation on this application. Any modifications made within the scope of the claims of this application are protected by the claims.

[0060] Unless otherwise stated, all raw materials and reagents used in the following examples are commercially available or can be prepared by known methods. Experimental methods not specifying particular conditions in the examples were performed under conventional conditions, such as those described in literature, books, or methods recommended by the manufacturer.

[0061] The neutral protease was purchased from Xiasheng (Beijing) Biotechnology Development Co., Ltd., model number FDG-2230.

[0062] The alkaline protease was purchased from Xiasheng (Beijing) Biotechnology Development Co., Ltd., model number FDY-2241.

[0063] The trypsin was purchased from Xiasheng (Beijing) Biotechnology Development Co., Ltd., model number FDG-2280.

[0064] Example 1

[0065] In this embodiment, neutral protease and collagen were used to prepare modified hydroxytyrosol.

[0066] (1) Add 0.5 g of hydroxytyrosol (HT), 0.5 g of neutral protease and 0.5 g of collagen to 10 ml of water to form solution A;

[0067] (2) Use acetic acid to adjust the pH of solution A to 6.5, thus forming solution B;

[0068] (3) Enzymatic hydrolysis was carried out by magnetic stirring at 45℃ for 5 hours to form solution C;

[0069] (4) After adding 2 mL of ethyl acetate to solution C, centrifuge the solution to deposit the enzyme preparation at the bottom, the water layer in the middle, and the ethyl acetate containing hydroxytyrosol at the top.

[0070] (5) The ethyl acetate layer obtained by centrifugation was collected and dried with anhydrous sodium sulfate to obtain modified hydroxytyrosol.

[0071] Example 2

[0072] In this embodiment, neutral protease and collagen were used to prepare modified hydroxytyrosol.

[0073] The preparation method is similar to that in Example 1, except that in step (1), 0.5 g of hydroxytyrosol (HT), 0.5 g of neutral protease and 1 g of collagen are added to 10 ml of water to form solution A.

[0074] Example 3

[0075] In this embodiment, modified hydroxytyrosol was prepared using trypsin and collagen.

[0076] The preparation method is similar to that in Example 1, except that in step (1), 0.5 g of hydroxytyrosol (HT), 0.5 g of trypsin and 0.5 g of collagen are added to 10 ml of water to form solution A.

[0077] Example 4

[0078] In this embodiment, modified hydroxytyrosol was prepared using neutral protease, alkaline protease, and collagen.

[0079] The preparation method is similar to that in Example 1, except that in step (1), 0.5 g of hydroxytyrosol (HT), 0.25 g of neutral protease, 0.25 g of alkaline protease and 0.5 g of collagen are added to 10 ml of water to form solution A.

[0080] Example 5

[0081] In this embodiment, modified hydroxytyrosol was prepared using neutral protease, trypsin, and collagen.

[0082] The preparation method is similar to that in Example 1, except that in step (1), 0.5 g of hydroxytyrosol (HT), 0.25 g of neutral protease, 0.25 g of trypsin and 0.5 g of collagen are added to 10 ml of water to form solution A.

[0083] Example 6

[0084] In this embodiment, modified hydroxytyrosol was prepared using trypsin, alkaline protease, and collagen.

[0085] The preparation method is similar to that in Example 1, except that in step (1), 0.5 g of hydroxytyrosol (HT), 0.25 g of trypsin, 0.25 g of alkaline protease and 0.5 g of collagen are added to 10 ml of water to form solution A.

[0086] Example 7

[0087] In this embodiment, modified hydroxytyrosol was prepared using neutral protease, alkaline protease, trypsin, and collagen.

[0088] The preparation method is similar to that in Example 1, except that in step (1), 0.5 g of hydroxytyrosol (HT), 0.17 g of neutral protease, 0.17 g of alkaline protease, 0.17 g of trypsin and 0.5 g of collagen are added to 10 ml of water to form solution A.

[0089] Example 8

[0090] In this embodiment, neutral protease and collagen were used to prepare modified hydroxytyrosol.

[0091] The preparation method is similar to that in Example 1, except that in step (1), 0.5 g of hydroxytyrosol (HT), 1 g of neutral protease and 1 g of collagen are added to 10 ml of water to form solution A.

[0092] Example 9

[0093] In this embodiment, neutral protease and collagen were used to prepare modified hydroxytyrosol.

[0094] The preparation method is similar to that in Example 1, except that in step (1), 0.5 g of hydroxytyrosol (HT), 0.1 g of neutral protease and 0.1 g of collagen are added to 10 ml of water to form solution A.

[0095] Example 10

[0096] In this embodiment, neutral protease and collagen were used to prepare modified hydroxytyrosol.

[0097] The preparation method is similar to that in Example 1, except that in step (2), acetic acid is used to adjust the pH of solution A to 5, forming solution B.

[0098] Example 11

[0099] In this embodiment, neutral protease and collagen were used to prepare modified hydroxytyrosol.

[0100] The preparation method is similar to that in Example 1, except that in step (2), acetic acid is used to adjust the pH of solution A to 6, forming solution B.

[0101] Example 12

[0102] In this embodiment, neutral protease and collagen were used to prepare modified hydroxytyrosol.

[0103] The preparation method is similar to that in Example 1, except that in step (2), acetic acid is used to adjust the pH of solution A to 6.8 to form solution B.

[0104] Example 13

[0105] In this embodiment, neutral protease and collagen were used to prepare modified hydroxytyrosol.

[0106] The preparation method is similar to that in Example 1, except that in step (3), the enzymatic hydrolysis reaction is carried out by magnetic stirring at 40°C for 8 hours to form solution C.

[0107] Example 14

[0108] In this embodiment, neutral protease and collagen were used to prepare modified hydroxytyrosol.

[0109] The preparation method is similar to that in Example 1, except that in step (3), the enzymatic hydrolysis reaction is carried out by magnetic stirring at 50°C for 4 hours to form solution C.

[0110] Example 15

[0111] In this embodiment, neutral protease and collagen were used to prepare modified hydroxytyrosol.

[0112] The preparation method is similar to that in Example 1, except that in step (3), the enzymatic hydrolysis reaction is carried out by magnetic stirring at room temperature (23°C) for 3 hours to form solution C.

[0113] Comparative Example 1

[0114] This comparative example uses collagenase and collagen to prepare modified hydroxytyrosol.

[0115] The preparation method is similar to that in Example 1, except that in step (1), 0.5 g of hydroxytyrosol (HT), 0.5 g of collagenase and 0.5 g of collagen are added to 10 ml of water to form solution A.

[0116] Comparative Example 2

[0117] This comparative example uses protease and yeast peptide to prepare modified hydroxytyrosol.

[0118] The preparation method is similar to that in Example 1, except that in step (1), 0.5 g of hydroxytyrosol (HT), 0.5 g of collagenase and 0.5 g of yeast peptide are added to 10 ml of water to form solution A.

[0119] Comparative Example 3

[0120] In this embodiment, modified hydroxytyrosol was prepared using alkaline protease and collagen.

[0121] The preparation method is similar to that in Example 1, except that in step (1), 0.5 g of hydroxytyrosol (HT), 0.5 g of alkaline protease and 0.5 g of collagen are added to 10 ml of water to form solution A.

[0122] Performance testing

[0123] I. Infrared Spectroscopy Detection

[0124] Infrared spectroscopy was performed on both the unmodified hydroxytyrosol and the prepared modified hydroxytyrosol, and the results are as follows: Figure 1 and Figure 2 As shown. Figure 1 The infrared spectrum of unmodified hydroxytyrosol. Figure 2 The infrared spectrum of the modified hydroxytyrosol obtained in Example 1 is shown.

[0125] contrast Figure 1 and Figure 2 The stretching vibration frequency of the OH group of unmodified hydroxytyrosol is 3375 cm⁻¹. -1 The stretching vibration frequency of the modified hydroxytyrosol (OH) is 3386 cm⁻¹. -1 The modified sample showed a significant red shift in peak values ​​compared to the unmodified raw material. The in-plane bending vibrations of the CH groups of certain aromatic rings are typically between 1000-1300 cm⁻¹. -1 Therefore, in unmodified hydroxytyrosol, at 1284 cm⁻¹ -1 Absorption peaks were also observed, but after modification with neutral protease and collagen catalysis, CN or NH peaks were clearly expressed, with the absorption peak at 1284 cm⁻¹. -1 There is a stronger expression at this point. As can be seen from the infrared spectrum, the amino group was successfully grafted onto hydroxytyrosol, achieving successful modification of hydroxytyrosol by neutral protease and collagen.

[0126] II. Mass Spectrometry Detection

[0127] The modified hydroxytyrosol obtained was analyzed by mass spectrometry.

[0128] Figure 3 The mass spectrum of the modified hydroxytyrosol prepared in Example 1 is shown. Figure 3As shown, during the modification of hydroxytyrosol by collagen and protease, collagen is broken down into amino acid monomers such as glycine, proline, and hydroxyproline under the enzymatic action of protease. These amino acid monomers then graft onto hydroxytyrosol for modification. Simultaneously, undigested collagen fragments also graft onto hydroxytyrosol as characteristic collagen fragments. Figure 3 As shown, the m / z of one of the ions in the modified hydroxytyrosol is 311. Since the detection method is positively charged mode (M+H), this indicates that its molecular mass is 310, which is consistent with the actual relative molecular mass. The molecular formula is C. 15 H 22 N2O5 2+ The other modified hydroxytyrosol ion has an m / z of 437. Since the detection method is positively charged mode (M+H), this indicates a molecular weight of 436, which matches the actual relative molecular weight. Its molecular formula is C2. 20 H 26 N3O8 3- .

[0129] III. DPPH Scavenging Experiment

[0130] Equal volumes of distilled water were added to prepare 0.1 mg / mL samples of unmodified hydroxytyrosol and the samples prepared in the examples. 2 mL of each hydroxytyrosol sample solution was added to 2 mL of DPPH (0.1 mM, methanol-soluble) solution, mixed thoroughly, and incubated at room temperature in the dark for 30 min. The absorbance was measured at 517 nm. Distilled water was used as the blank group, and vitamin C (Vc) was used as the positive control. All samples were measured in triplicate, and the average value was calculated. The results are shown in Table 1. The scavenging ability is expressed as the scavenging rate, which was calculated using the following formula:

[0131]

[0132] Among them, A 空白 The absorbance of the blank group (2 mL of distilled water was used to replace the sample);

[0133] A 样品 The absorbance of the sample solution;

[0134] A 对照 The absorbance is calculated using 2 mL of methanol instead of DPPH.

[0135] Table 1

[0136]

[0137] As shown in Table 1, the DPPH free radical scavenging data of modified hydroxytyrosol exhibit varying degrees of improvement in DPPH free radical scavenging compared to unmodified hydroxytyrosol. The modified hydroxytyrosol prepared using neutral protease and collagen in Example 1 showed the greatest increase in scavenging rate. This may be because the modified hydroxytyrosol carries more active charges, resulting in higher efficiency in free radical quenching and enhanced antioxidant capacity, thus achieving the goal of enhanced antioxidant effects.

[0138] IV. ABTS Removal Experiment

[0139] Preparation of ABTS free radical stock solution: Mix 10 mL of 7.4 mol / L ABTS solution and 10 mL of 2.6 mmol / L K2S2O8 solution, and oxidize at room temperature in the dark for 12 h to obtain the stock solution. Before use, the ABTS stock solution was diluted with PBS (0.2 M, pH=7.4) to achieve an absorbance of 0.7 ± 0.02 at 734 nm. Add an equal volume of distilled water to prepare the unmodified hydroxytyrosol and the sample prepared in the examples to a concentration of 0.1 mg / mL. Take 2 mL of ABTS solution, add 2 mL of sample solutions from different groups, mix, and let stand at room temperature for 6 min. Measure the absorbance at 734 nm. Use ascorbic acid (Vc) as a positive control. All samples were measured in triplicate, and the average value was calculated. The results are shown in Table 2. The ABTS scavenging ability is expressed as the scavenging rate, calculated using the following formula:

[0140]

[0141] Among them, A 空白 The absorbance of the blank group (2 mL of distilled water was used to replace the sample);

[0142] A 样品 The absorbance of the sample solution;

[0143] A 对照 2 mL of sample control (2 mM 0.2M buffer instead of ABTS).

[0144] Table 2

[0145]

[0146] As shown in Table 2, the ABTS radical scavenging data indicate that the modified hydroxytyrosol demonstrates improved ABTS radical scavenging performance compared to the unmodified hydroxytyrosol. Furthermore, considering the DPPH radical scavenging rate data, the modified hydroxytyrosol obtained in Example 1 exhibits the best antioxidant effect.

[0147] V. Stability Test

[0148] Weigh 0.01 g of unmodified hydroxytyrosol and the modified hydroxytyrosol obtained in Example 1, respectively, and dissolve them in 10 mL of pure water until completely dissolved. Place them in a 45°C oven overnight (12 h) and observe the color change of the samples. The results are as follows: Figure 4 As shown.

[0149] Depend on Figure 4 The experimental results show that the stability of the modified hydroxytyrosol was significantly improved. After being placed in an oven at 45°C overnight (12h), no obvious color change was observed, while the unmodified hydroxytyrosol showed obvious color change. The reason may be that after the modified hydroxytyrosol was grafted with amino groups, the active charge it carried improved the internal color change mechanism of the hydroxytyrosol molecule and improved the stability of the sample.

[0150] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0151] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention should be determined by the appended claims, and the specification and drawings can be used to interpret the content of the claims.

Claims

1. A method for preparing modified hydroxytyrosol for use in cosmetics or food, characterized in that, The process includes the following steps: hydroxytyrosol, protease, and collagen are mixed and reacted in the presence of an acidic solvent; the reaction product is mixed with an extractant; the mixture is centrifuged to separate the layers; the upper layer is collected to prepare the modified hydroxytyrosol. The protease includes one or both of neutral proteases and trypsin.

2. The method for preparing modified hydroxytyrosol for cosmetics or food according to claim 1, characterized in that, The mass ratio of hydroxytyrosol to collagen is 1:(1~2).

3. The method for preparing modified hydroxytyrosol for cosmetics or food according to claim 1, characterized in that, The mass ratio of hydroxytyrosol to the protease is 1:(1~2).

4. The method for preparing modified hydroxytyrosol for cosmetics or food according to any one of claims 1 to 3, characterized in that, The solvent has a pH value of 6 to 6.

8.

5. The method for preparing modified hydroxytyrosol for cosmetics or food according to any one of claims 1 to 3, characterized in that, The reaction temperature is 40℃~50℃, and the reaction time is 4h~8h.

6. The method for preparing modified hydroxytyrosol for cosmetics or food according to any one of claims 1 to 3, characterized in that, One or more of the following conditions must be met: (1) The extractant includes ethyl acetate; (2) The mass-volume ratio of the hydroxytyrosol to the extractant is 1 g: (2~4) mL.

7. The method for preparing modified hydroxytyrosol for cosmetics or food according to any one of claims 1 to 3, characterized in that, One or more of the following conditions must be met: (1) The solvent is water; (2) The mass-to-volume ratio of the hydroxytyrosol to the solvent is 1 g: (8~15) mL; (3) In the centrifugation layering step, centrifuge at 5000rpm~8000rpm for 5min~8min.

8. A modified hydroxytyrosol, characterized in that, It is prepared by the preparation method according to any one of claims 1 to 7.

9. The use of the modified hydroxytyrosol according to claim 8 in the preparation of products with free radical scavenging effects; The products mentioned include cosmetics or food.

10. A cosmetic or food product, characterized in that, Contains the modified hydroxytyrosol as described in claim 8.

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