Modified hydroxytyrosol, its preparation methods and applications, and cosmetics

Abstract

This application provides modified hydroxytyrosol, its preparation method, and its application in cosmetics, belonging to the field of cosmetic technology. This application involves mixing crude hydroxytyrosol, cysteine, and water, then adding peroxidase and hydrogen peroxide to react and generate modified hydroxytyrosol. The preparation method is green and safe, and the flavor of the modified hydroxytyrosol is significantly improved, with a substantial increase in pleasant substances, effectively improving the unpleasant flavors such as irritation and bitterness of the crude hydroxytyrosol raw material.

Description

Technical Field

[0001] This application relates to the field of cosmetic formulation technology, and in particular to modified hydroxytyrosol, its preparation method and application, and cosmetics. Background Technology

[0002] Hydroxytyrosol possesses exceptional antioxidant capabilities, making it one of the most powerful natural antioxidants found in nature. Currently, hydroxytyrosol can be obtained from various sources, primarily through plant extraction, microbial fermentation, biotransformation, and chemical synthesis. Among these, chemical synthesis is widely used due to its lower cost and suitability for large-scale production. However, chemical synthesis involves various organic reagents, and these reagents introduced during extraction or purification result in a strong, pungent, and bitter taste in hydroxytyrosol, significantly impacting its application in cosmetic formulations.

[0003] Therefore, how to modify hydroxytyrosol to improve the unpleasant flavor caused by organic reagents in hydroxytyrosol raw materials has become an urgent technical problem to be solved. Summary of the Invention

[0004] Based on this, the main purpose of this application is to provide modified hydroxytyrosol, its preparation method and application in cosmetics, so as to improve the flavor of the raw materials of modified hydroxytyrosol and avoid the flavor of the raw materials affecting the application and development of hydroxytyrosol-related products.

[0005] The first aspect of this application provides a method for preparing modified hydroxytyrosol, comprising the following steps:

[0006] Crude hydroxytyrosol, cysteine, and water were mixed, and then peroxidase and hydrogen peroxide were added to react and produce modified hydroxytyrosol.

[0007] The modified hydroxytyrosol includes compounds having the structure shown below:

[0008] .

[0009] In some embodiments, the step of adding peroxidase and hydrogen peroxide to carry out the reaction includes the following conditions: the reaction temperature is 40°C-50°C; the reaction time is 4h-5h.

[0010] In some embodiments, the mass ratio of the crude hydroxytyrosol to the cysteine ​​is 1:(1-2).

[0011] In some embodiments, the crude hydroxytyrosol comprises hydroxytyrosol and volatile impurities; the volatile impurities include n-butanol and tetramethyl silicate.

[0012] In some embodiments, the mass ratio of the crude hydroxytyrosol, the peroxidase, and the hydrogen peroxide is 500:(40-80):(20.4-47.6).

[0013] In some embodiments, the peroxidase is horseradish peroxidase; the activity concentration of the peroxidase is 150 U / g-180 U / g.

[0014] In some embodiments, after reacting with peroxidase and hydrogen peroxide, the method further includes the step of extracting the modified hydroxytyrosol with an organic solvent.

[0015] A second aspect of this application provides a modified hydroxytyrosol, prepared by the method for preparing modified hydroxytyrosol described in the first aspect, wherein the modified hydroxytyrosol comprises a volatile component and a compound having the structure shown below:

[0016] .

[0017] In some embodiments, the volatile component includes at least one of aromatic compounds, pyranone compounds, sugar compounds, and urea.

[0018] The third aspect of this application provides the use of modified hydroxytyrosol prepared by the method described in the first aspect or the modified hydroxytyrosol described in the second aspect in the preparation of cosmetics.

[0019] In a fourth aspect of this application, a cosmetic product is provided, comprising modified hydroxytyrosol prepared by the method described in the first aspect or modified hydroxytyrosol described in the second aspect.

[0020] Compared with traditional technologies, this application has at least the following beneficial effects:

[0021] This application modifies crude hydroxytyrosol using cysteine ​​in a system containing water, peroxidase, and hydrogen peroxide. The preparation method is green and safe, and the modified hydroxytyrosol has a significantly improved flavor with a substantial increase in pleasant substances, effectively improving the unpleasant flavors such as irritation and bitterness of the crude hydroxytyrosol raw material. Attached Figure Description

[0022] To better describe and illustrate the embodiments or examples provided in this application, reference may be made to one or more accompanying drawings. Additional details or examples used to describe the drawings should not be considered as limiting the scope of any of the disclosed applications, the currently described embodiments or examples, or the best mode of conduct of these applications as currently understood. Furthermore, the same reference numerals denote the same parts throughout the drawings.

[0023] Figure 1 The infrared spectrum of crude hydroxytyrosol in Example 1;

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

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

[0026] Figure 4 The total ion chromatogram of volatile components in the crude hydroxytyrosol from Example 1 is shown.

[0027] Figure 5 The total ion chromatogram of the volatile components of the modified hydroxytyrosol in Example 1 is shown.

[0028] Figure 6 The fingerprint spectra of volatile components in samples 1, 2 and 3 are shown. Detailed Implementation

[0029] The present application will be further described in detail below with reference to the embodiments and examples. These embodiments and examples are for illustrative purposes only and are not intended to limit the scope of the present application. The purpose of providing these embodiments and examples is to enable a more thorough and comprehensive understanding of the disclosure of the present application. It should also be understood that the present application can be implemented in many different forms and is not limited to the embodiments and examples described herein. Those skilled in the art can make various modifications or alterations without departing from the spirit of the present application, and the equivalent forms obtained also fall within the protection scope of the present application. Furthermore, numerous specific details are set forth in the following description to provide a fuller understanding of the present application. It should be understood that the present application can be implemented without one or more of these details.

[0030] 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 application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0031] To address the unpleasant flavor issues caused by organic reagents introduced during the extraction or purification of hydroxytyrosol, this application modifies crude hydroxytyrosol using cysteine ​​in a system containing water, peroxidase, and hydrogen peroxide. The preparation method is green and safe, and the modified hydroxytyrosol obtained has a significantly improved flavor with a substantial increase in pleasant substances, effectively improving the unpleasant flavors such as irritation and bitterness of the crude hydroxytyrosol raw material.

[0032] The first aspect of this application provides a method for preparing modified hydroxytyrosol, comprising the following steps:

[0033] Crude hydroxytyrosol, cysteine, and water were mixed, and then peroxidase and hydrogen peroxide were added to react and produce modified hydroxytyrosol.

[0034] The modified hydroxytyrosol includes compounds having the structure shown below:

[0035] .

[0036] This application utilizes a system containing water, peroxidase, and hydrogen peroxide to modify crude hydroxytyrosol using cysteine. Compared to the raw crude hydroxytyrosol, the modified hydroxytyrosol exhibits significantly improved flavor, with a substantial increase in pleasant compounds, effectively mitigating the harshness and bitterness of the crude hydroxytyrosol. Specifically, during the preparation of the modified hydroxytyrosol, cysteine ​​is used to graft the hydroxytyrosol, thereby separating it from the organic reagents in the crude hydroxytyrosol. Simultaneously, new functional groups are introduced into the structure of the hydroxytyrosol, giving it a superior odor and thus improving the unpleasant odor of the raw crude hydroxytyrosol.

[0037] In some embodiments, the crude hydroxytyrosol comprises hydroxytyrosol and volatile impurities, including n-butanol and tetramethyl silicate. The mixed n-butanol and tetramethyl silicate in the crude hydroxytyrosol have a distinctly pungent odor, resulting in a generally pungent taste in the crude hydroxytyrosol.

[0038] In some embodiments, the crude hydroxytyrosol comprises hydroxytyrosol and volatile impurities, including n-butanol, 2,2-dimethoxybutane, tetramethyl silicate, butyl acetate, 2,6-di-tert-butyl-p-cresol, 2-cyanobiphenyl, 3,4-dihydroxyphenylethanol, and 2,1,3-benzothiadiazole.

[0039] In some embodiments, the step of adding peroxidase and hydrogen peroxide to carry out the reaction includes the following conditions: the reaction temperature is 40℃-50℃, which can be 40℃, 41℃, 42℃, 43℃, 44℃, 45℃, 46℃, 47℃, 48℃, 49℃ or 50℃; the reaction time is 4h-5h, which can be 4h, 4.2h, 4.5h, 4.8h or 5h.

[0040] In some embodiments, the mass ratio of the crude hydroxytyrosol to the cysteine ​​is 1:(1-2), which can be 1:1, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9 or 1:2.

[0041] In some embodiments, the conditions for mixing crude hydroxytyrosol, peroxidase, hydrogen peroxide, and water include: a temperature of 35°C-40°C, which can be 35°C, 36°C, 37°C, 38°C, 39°C, or 40°C; and a time of 1h-2h, which can be 1h, 1.2h, 1.5h, 1.8h, or 2h.

[0042] In some embodiments, the mass ratio of the crude hydroxytyrosol, the peroxidase, and the hydrogen peroxide is 500:(40-80):(20.4-47.6), which can be 500:40:20.4, 500:40:47.6, 500:60:20.4, 500:60:47.6, 500:40:34, 500:60:34, 500:80:20.4, or 500:80:47.6.

[0043] In some embodiments, the peroxidase is horseradish peroxidase (HRP); the activity concentration of the peroxidase is 150 U / g-180 U / g.

[0044] In some embodiments, the hydrogen peroxide is provided by hydrogen peroxide solution at a concentration of 0.2 mol / L - 0.6 mol / L, which can be 0.2 mol / L, 0.3 mol / L, 0.4 mol / L, 0.5 mol / L, or 0.6 mol / L.

[0045] This application describes a process where, in a system containing water, peroxidase, hydrogen peroxide, and cysteine, hydroxytyrosol in crude hydroxytyrosol is oxidized to a quinone intermediate by peroxidase and hydrogen peroxide. The quinone intermediate then undergoes an addition reaction with cysteine, thereby achieving graft modification of cysteine ​​onto hydroxytyrosol to generate modified hydroxytyrosol. The reaction formula is as follows:

[0046] .

[0047] In some embodiments, after reacting with peroxidase and hydrogen peroxide, the method further includes the step of extracting the modified hydroxytyrosol with an organic solvent.

[0048] In some embodiments, the step of extracting the modified hydroxytyrosol with an organic solvent includes ethyl acetate.

[0049] In some embodiments, the method for preparing modified hydroxytyrosol includes the following steps:

[0050] The crude hydroxytyrosol, cysteine, and water are mixed, and then peroxidase and hydrogen peroxide are added to react. The modified hydroxytyrosol is then extracted with an organic solvent. The volume ratio of the organic solvent to the water is 1:(3-6), which can be 1:3, 1:4, 1:5, or 1:6.

[0051] In some embodiments, after extracting the modified hydroxytyrosol with an organic solvent, the step of drying the extracted modified hydroxytyrosol is further included.

[0052] In some embodiments, the drying conditions include drying with anhydrous sodium sulfate. In some embodiments, the dried modified hydroxytyrosol is a yellow liquid.

[0053] In some embodiments, the modified hydroxytyrosol further includes volatile components; the volatile components include at least one of aromatic compounds, pyranone compounds, sugar compounds, and urea.

[0054] In some embodiments, the aromatic compound includes at least one selected from benzyl alcohol, homovanillin, 2,6-di-tert-butylnaphthalene, catechol, 4-(2,2,4-trimethyl-3,4-dihydro-2H-chromene)phenol, 4-ethylphenylphosphine, 4-(2-allyl)-catechol, 2,6-di-tert-butyl-p-cresol, and chromenepyridine amine derivatives.

[0055] In some embodiments, the pyranone compounds include 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one.

[0056] In some embodiments, the carbohydrate compound includes D-allose.

[0057] In some embodiments, the volatile components include benzyl alcohol, urea, 2,6-di-tert-butylnaphthalene, catechol, chromonepyridine amine derivatives, 4-(2,2,4-trimethyl-3,4-dihydro-2H-chromene)phenol, 4-ethylphenylphosphine, 4-(2-allyl)-catechol, 2,6-di-tert-butyl-p-cresol, D-aloose, homovanillin, and 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one.

[0058] A second aspect of this application provides a modified hydroxytyrosol, prepared by the method for preparing modified hydroxytyrosol described in the first aspect, wherein the modified hydroxytyrosol comprises a volatile component and a compound having the structure shown below:

[0059] .

[0060] In some embodiments, the volatile component includes at least one of aromatic compounds, pyranone compounds, sugar compounds, and urea.

[0061] In some embodiments, the aromatic compound includes at least one selected from benzyl alcohol, homovanillin, 2,6-di-tert-butylnaphthalene, catechol, 4-(2,2,4-trimethyl-3,4-dihydro-2H-chromene)phenol, 4-ethylphenylphosphine, 4-(2-allyl)-catechol, 2,6-di-tert-butyl-p-cresol, and chromenepyridine amine derivatives.

[0062] In some embodiments, the pyranone compounds include 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one.

[0063] In some embodiments, the carbohydrate compound includes D-allose.

[0064] In some embodiments, the volatile components include benzyl alcohol, urea, 2,6-di-tert-butylnaphthalene, catechol, chromonepyridine amine derivatives, 4-(2,2,4-trimethyl-3,4-dihydro-2H-chromene)phenol, 4-ethylphenylphosphine, 4-(2-allyl)-catechol, 2,6-di-tert-butyl-p-cresol, D-aloose, homovanillin, and 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one.

[0065] In some embodiments, the modified hydroxytyrosol is a yellow liquid.

[0066] The third aspect of this application provides the use of modified hydroxytyrosol prepared by the method described in the first aspect or the modified hydroxytyrosol described in the second aspect in the preparation of cosmetics.

[0067] Hydroxytyrosol possesses excellent antioxidant capabilities and primarily functions as a free radical scavenger in cosmetics, with its core active groups being its catechol structure and phenolic hydroxyl groups. Therefore, compared to crude hydroxytyrosol, the modified hydroxytyrosol prepared in this application not only improves the unpleasant flavor of the crude raw material but also retains its free radical scavenging function, thus enhancing its application value in cosmetics.

[0068] In a fourth aspect of this application, a cosmetic product is provided, comprising modified hydroxytyrosol prepared by the method described in the first aspect or modified hydroxytyrosol described in the second aspect.

[0069] The embodiments of this application will be described in detail below with reference to examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of this application. For experimental methods in the following embodiments where specific conditions are not specified, please refer to the guidelines given in this application, or follow experimental manuals or conventional conditions in the art, or follow the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0070] As an example, the raw materials used in the embodiments and comparative examples of this application are as follows:

[0071] Crude hydroxytyrosol: purchased from Chongqing Boteng Pharmaceutical Technology Co., Ltd., production batch number 2505002;

[0072] Horseradish peroxidase: purchased from Shanghai Aladdin Biochemical Technology Co., Ltd., model number H597642, with an activity concentration of 150 U / mg;

[0073] Lipase: Purchased from Xiasheng (Beijing) Biotechnology Development Co., Ltd., model FDG-2208, activity concentration 100,000 U / g.

[0074] Example 1

[0075] The specific preparation method of modified hydroxytyrosol is as follows:

[0076] 500 mg of crude hydroxytyrosol (HT) and 800 mg of cysteine ​​were dissolved completely in 10 mL of water. Then, 80 mg of horseradish peroxidase (HRP) and 3.5 mL of hydrogen peroxide (0.4 mol / L hydrogen peroxide concentration, 47.6 mg of hydrogen peroxide added to 3.5 mL of hydrogen peroxide) were added and the mixture was stirred at 40 °C and 300 rpm for 4 h. After the reaction was completed, 2 mL of ethyl acetate was added for extraction, the ethyl acetate phase was collected, dried over anhydrous sodium sulfate, and filtered to obtain modified hydroxytyrosol (yellow liquid).

[0077] Example 2

[0078] The preparation method of modified hydroxytyrosol in Example 2 is basically the same as that in Example 1, except that "800mg cysteine" is replaced with "600mg cysteine"; and modified hydroxytyrosol is prepared according to the method in Example 1.

[0079] Example 3

[0080] The preparation method of modified hydroxytyrosol in Example 3 is basically the same as that in Example 1, except that "mixing and stirring at 40°C and 300 rpm for 4 hours" is replaced with "mixing and stirring at 50°C and 300 rpm for 4 hours"; modified hydroxytyrosol is prepared according to the method in Example 1.

[0081] Example 4

[0082] The preparation method of modified hydroxytyrosol in Example 4 is basically the same as that in Example 1, except that "mixing and stirring at 40°C and 300 rpm for 4 hours" is replaced with "mixing and stirring at 40°C and 300 rpm for 5 hours"; modified hydroxytyrosol is prepared according to the method in Example 1.

[0083] Comparative Example 1

[0084] The preparation method of modified hydroxytyrosol in Comparative Example 1 is basically the same as that in Example 1, except that "80 mg horseradish peroxidase and 3.5 mL hydrogen peroxide (hydrogen peroxide concentration of 0.4 mol / L, and the amount of hydrogen peroxide added in 3.5 mL hydrogen peroxide is 47.6 mg)" is replaced with "80 mg lipase"; modified hydroxytyrosol is prepared according to the method of Example 1.

[0085] Comparative Example 2

[0086] The preparation method of modified hydroxytyrosol in Comparative Example 2 is basically the same as that in Example 1, except that "cysteine" is replaced with an equal amount of "glycine"; and modified hydroxytyrosol is prepared according to the method in Example 1.

[0087] Experimental Example 1

[0088] (1) Structural identification of modified hydroxytyrosol and crude hydroxytyrosol in Example 1

[0089] The modified hydroxytyrosol and crude hydroxytyrosol from Example 1 were subjected to infrared spectroscopy and mass spectrometry analysis. The results are as follows: Figures 1-3 As shown.

[0090] in, Figure 1 The image shows the infrared spectrum of the crude hydroxytyrosol from Example 1. Figure 2 The infrared spectrum of the modified hydroxytyrosol in Example 1 is shown for comparison. Figures 1-2 It can be seen that, Figure 1 The stretching vibration frequency of the OH group in hydroxytyrosol is 3375.19 cm⁻¹. -1 ,and Figure 2 The stretching vibration frequency of the OH group in modified hydroxytyrosol is 3416.82 cm⁻¹. -1 This indicates that the OH elution peak of modified hydroxytyrosol shows a significant red shift compared to unmodified hydroxytyrosol. Compared to the infrared spectrum of crude hydroxytyrosol, the modified hydroxytyrosol exhibits a significantly higher elution peak at 2069.02 cm⁻¹. -1 A distinct absorption peak appears at [value missing], and based on the peak position, it is likely the stretching vibration peak of the C=O bond in carbonyl sulfide (O=C=S). Meanwhile, the C=S bond typically occurs between 700-590 cm⁻¹. -1 A weak intensity of expression was observed at 629.56 cm⁻¹. Based on the peak position, the modified hydroxytyrosol was determined to be present at this value.-1 The absorption peak at 1631.90 cm⁻¹ is a stretching vibration peak of the CS bond. The appearance of these characteristic peaks proves that cysteine ​​has been successfully grafted onto hydroxytyrosol via a thiol group, indicating that the modified hydroxytyrosol contains a CS bond. Furthermore, compared to hydroxytyrosol, the modified hydroxytyrosol exhibits a higher absorption peak at 1631.90 cm⁻¹. -1 A distinct absorption peak appears at [location missing], which corresponds to the characteristic peak of the stretching vibration of C=O in amides or carboxylic acids (1680-1630 cm⁻¹). -1 The results show a high degree of agreement, and the appearance of this absorption peak indicates that the cysteine-hydroxytyrosol graft product was successfully synthesized.

[0091] Figure 3 The mass spectrum of the modified hydroxytyrosol in Example 1 is shown below. Figure 3 It can be seen that the m / z of the major ion of modified hydroxytyrosol is 274. Since the detection method is positively charged mode (M+H), the molecular weight of modified hydroxytyrosol is 273. Furthermore, based on the reaction mechanism, it can be deduced that the reaction of hydroxytyrosol with cysteine, under the catalytic system of hydrogen peroxide and peroxidase (horseradish peroxidase), results in a molecular weight of 273 for modified hydroxytyrosol, with the molecular formula C273. 11 H 15 NO5S is consistent with the results of infrared spectroscopy detection.

[0092] Combination Figures 1-3 As can be seen, in Example 1, hydroxytyrosol in crude hydroxytyrosol was oxidized to a quinone intermediate by peroxidase and peroxide in a system containing water, peroxidase, hydrogen peroxide, and cysteine. The quinone intermediate then underwent an addition reaction with cysteine, thereby achieving graft modification of cysteine ​​onto hydroxytyrosol to generate modified hydroxytyrosol. Among these, the ortho-benzoquinone structure in the quinone intermediate has a lower electron cloud density at the C-5 position, exhibiting stronger electrophilicity and making it more prone to generating graft products. The reaction formula is as follows:

[0093] .

[0094] (2) Volatile component analysis of modified hydroxytyrosol and crude hydroxytyrosol in Example 1

[0095] GCMS (Gas Chromatography-Mass Spectrometry) provides high-resolution mass spectra, which helps in the accurate identification of compounds. Using GCMS, the mass spectrometry data of the components were automatically retrieved from the NIST20.L spectral library, and the search results were manually verified. The names and CAS numbers of the compounds formed by each peak position were checked. The main volatile components in the modified hydroxytyrosol and crude hydroxytyrosol of Example 1 were analyzed, and the relative content (relative volume percentage) of the main volatile components was calculated using the peak area normalization method.

[0096] The results are shown in Tables 1-2 and 2. Figures 4-5 .

[0097] Table 1. Main volatile components and relative contents of crude hydroxytyrosol in Example 1

[0098]

[0099] Table 2. Main volatile components and relative contents of the modified hydroxytyrosol prepared in Example 1

[0100]

[0101] Table 1 shows that, apart from hydroxytyrosol, the main volatile components in the crude hydroxytyrosol accounted for 99.9% of the total volatile components. The main volatile components of hydroxytyrosol were as follows in relative amounts: 1.74% n-butanol, 0.05% 2,2-dimethoxybutane, 0.09% tetramethyl silicate, 0.17% butyl acetate, 0.43% 2,6-di-tert-butyl-p-cresol, 0.15% 2-cyanobiphenyl, and 1.23% 2,1,3-benzothiadiazole. Among them, n-butanol had a distinct bitter taste, tetramethyl silicate had a distinct pungent odor, and although butyl acetate itself had a fruity aroma, it was easily masked by the pungent odor. The remaining volatile components each had their own characteristic odor, and the overall mixture presented a relatively pungent taste.

[0102] Table 2 shows that, apart from the modified hydroxytyrosol itself, the main volatile components in the modified hydroxytyrosol prepared in Example 1 accounted for 99.9% of the total volatile components. The main volatile components in the modified hydroxytyrosol were as follows (in relative amounts): 4.78% benzyl alcohol, 0.27% urea, 0.97% 2,6-di-tert-butylnaphthalene, 0.13% catechol, 0.11% chromonepyridine amine derivatives, 0.42% 4-(2,2,4-trimethyl-3,4-dihydro-2H-chromone)phenol, 0.25% 4-ethylphenylphosphine, 0.37% 4-(2-allyl)-catechol, 0.2% 2,6-di-tert-butyl-p-cresol, 0.49% D-allose, 0.89% homovanillin, and 0.74% [other components not specified in the original text]. 2,3-Dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one; wherein benzyl alcohol has a light fragrance, 2,3-dihydro-3,5-dihydroxy-6-methyl-4(H)-pyran-4-one has the food fragrance of pyran compounds, and high vanillin has an aromatic odor.

[0103] Comparing Tables 1 and 2, it can be seen that after modification, hydroxytyrosol produces a variety of volatile components with pleasant odors, effectively improving the obvious irritation of the crude hydroxytyrosol raw material.

[0104] Experimental Example 2

[0105] GC-IMS (Gas Chromatography Ion Mobility Spectrometry) is characterized by rapid identification and comparison. Although its accuracy is lower than that of GCMS, it is good at performing rapid analysis of complex odors / volatiles in the form of "fingerprints". GC-IMS can be used to further compare and analyze the differences in volatile odors between modified hydroxytyrosol and crude hydroxytyrosol raw materials.

[0106] The modified hydroxytyrosol prepared in Example 1 was used as Sample 1, the crude hydroxytyrosol from Example 1 was used as Sample 2, and the modified hydroxytyrosol prepared in Comparative Example 1 was used as Sample 3. The flavor differences between Sample 1 and Sample 3 were analyzed by GC-IMS, and the specific methods are as follows:

[0107] Volatile organic compounds were detected using a Flavor Spec1H1-00053 GC-IMS system (manufactured by GAS GmbH, Germany). 1g of sample 1, 1g of sample 2, and 1g of sample 3 were weighed and placed in 20mL headspace vials, incubated at 50℃ for 20min, and then injected.

[0108] The GC-IMS instrument comes with VOCal software, two plugins, and GC×IMS LibrarySearch, which can analyze samples from different perspectives.

[0109] Among them, the VOCal software is used to view and analyze spectra, with each point in the graph representing a volatile organic compound; after establishing a standard curve, quantitative analysis can be performed; the Reporter plugin allows direct comparison of spectral differences between samples (two-dimensional top view and three-dimensional spectrum); the Gallery Plot plugin is used for fingerprint spectrum comparison, intuitively and quantitatively comparing the differences in volatile organic compounds between different samples.

[0110] To clearly identify and compare the specific volatile components that differ between Sample 1, Sample 2, and Sample 3, fingerprint spectra of all peaks were selected for rapid identification and comparison.

[0111] The fingerprint spectra of volatile components in samples 1, 2, and 3 are as follows: Figure 6 As shown, based on the content of volatile components in the three samples, the fingerprint spectra of the three samples are divided into three regions: Region A indicates that volatile components were detected in all three samples; Region B indicates that the content of volatile components in Sample 1 is higher than that in Sample 2 and Sample 3; Region C indicates that the content of volatile components in Sample 2 and Sample 3 is higher than that in Sample 1.

[0112] Figure 6 middle:

[0113] The volatile components marked in area A were detected in all three samples. Among them, 1-butanol had a fusel oil and whiskey odor; (Z)-2-penten-1-ol had a fruity aroma; butyl acetate had a light aroma; 2-methyl-2-propanol had a camphor odor; 2,3-diethyl-5-methylpyrazine had a coffee and pome aroma; and isopropyl acetate had a weak aroma.

[0114] The volatile components marked in region B are substances with higher content in sample 1 compared to samples 2 and 3. Among them, 1-octene-M and 1-octene-D have ester aromas; methyl isovalerate has herbal and fruity aromas; 1-propanethiol has a cabbage and natural gas aroma; 2-methyl-1-propylacetate has a fruity ester aroma; 2-acetyl-5-methylfuran has a musty and nutty aroma; 4-methyl-2-pentanone has a camphor aroma; (Z)-4-heptenal has a grassy and oily aroma; propyl butyrate has a fruity aroma; 2-ethyl-5-methylpyrazine has a coffee aroma; propyl propionate has a fruity aroma; 2-methylbutanal has a musty, cocoa, and phenolic aroma; dimethyl trisulfide has a minty and spicy aroma; and 2-ethyl-4-methylthiazole has a nutty aroma.

[0115] The substances marked in region C are the volatile components that are present in higher amounts in samples 2 and 3 compared to sample 1. Among them, acetic acid-M and acetic acid-D have an irritating odor; tetrahydrofuran has an ether-like odor; ethyl formate has a pungent, irritating odor and a pineapple-like fruity aroma; 1-propanol has a musty, slightly bitter, and astringent odor similar to topical rubbing alcohol; propionic acid has an irritating odor; 1-penten-3-one has a spicy, ethereal, peppery, garlic, mustard, and onion-like odor; and isopropanol has an ethanol-like odor.

[0116] The fingerprint data above show that the volatile components detected in Sample 1 are mainly pleasant aroma components, while the volatile components with higher content in Samples 2 and 3 are mainly irritating components. This indicates that the flavor of the modified hydroxytyrosol prepared in Example 1 is significantly improved compared with the crude hydroxytyrosol and the modified hydroxytyrosol in Comparative Example 1.

[0117] Experimental Example 3

[0118] Using crude hydroxytyrosol from Example 1, and modified hydroxytyrosol prepared in Examples 1-4 and Comparative Examples 1-2 as samples, eight sensory evaluators aged 19-25 years (4 males and 4 females) were selected to conduct a comprehensive evaluation of the samples, mainly focusing on odor. The relevant evaluation content and standards are shown in Table 3. Specifically, all sensory evaluators received one year of sensory training and were familiar with bitter, aromatic, pungent, sour, and full-bodied tastes. The evaluators conducted blind evaluations of the samples according to the evaluation standards and provided scores for each indicator.

[0119] Table 3. Sensory Evaluation Criteria for Odor

[0120]

[0121] The sensory evaluation results of each sample are shown in Table 4.

[0122] Table 4 Sensory evaluation results for each sample

[0123]

[0124] The results in Table 4 show that the modified hydroxytyrosol prepared in Examples 1-4 has a significantly improved flavor, with the modified hydroxytyrosol prepared in Example 1 having the best flavor.

[0125] In the evaluation of bitterness, compared with crude hydroxytyrosol and the modified hydroxytyrosol in Comparative Examples 1-2, the bitterness of the modified hydroxytyrosol in Examples 1-4 was significantly reduced, with Example 1 having the lowest bitterness.

[0126] In the aroma evaluation, compared with crude hydroxytyrosol and the modified hydroxytyrosol in Comparative Examples 1-2, the aroma of the modified hydroxytyrosol in Examples 1-4 was significantly improved, with Example 1 having the highest aroma.

[0127] In the evaluation of pungent taste, compared with crude hydroxytyrosol and the modified hydroxytyrosol in Comparative Examples 1-2, the pungent taste of the modified hydroxytyrosol in Examples 1-4 was significantly reduced, with Example 1 having the lowest pungent taste.

[0128] In the acidity evaluation, the acidity of the modified hydroxytyrosol in Example 1 was significantly reduced compared with crude hydroxytyrosol and the modified hydroxytyrosol in Comparative Examples 1-2.

[0129] 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.

[0130] The embodiments described above are merely illustrative of several implementation methods of this application, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this patent application should be determined by the appended claims.

Claims

1. A method for preparing modified hydroxytyrosol, characterized in that, Includes the following steps: Crude hydroxytyrosol, cysteine, and water were mixed, and then peroxidase and hydrogen peroxide were added to react and produce modified hydroxytyrosol. The modified hydroxytyrosol includes compounds having the structure shown below: 。 2. The method for preparing modified hydroxytyrosol according to claim 1, characterized in that, The reaction steps involving the addition of peroxidase and hydrogen peroxide include the following conditions: the reaction temperature is 40℃-50℃; the reaction time is 4h-5h.

3. The method for preparing modified hydroxytyrosol according to claim 1, characterized in that, It meets at least one of the following characteristics: (1) The mass ratio of the crude hydroxytyrosol to the cysteine ​​is 1:(1-2); (2) The crude hydroxytyrosol product includes hydroxytyrosol and volatile impurities; the volatile impurities include n-butanol and tetramethyl silicate.

4. The method for preparing modified hydroxytyrosol according to claim 1, characterized in that, The mass ratio of the crude hydroxytyrosol, the peroxidase, and the hydrogen peroxide is 500:(40-80):(20.4-47.6).

5. The method for preparing modified hydroxytyrosol according to claim 4, characterized in that, The peroxidase is horseradish peroxidase; The activity concentration of the peroxidase is 150 U / mg-180 U / mg.

6. The method for preparing modified hydroxytyrosol according to any one of claims 1-5, characterized in that, After reacting with peroxidase and hydrogen peroxide, the process further includes the following step: extracting the modified hydroxytyrosol with an organic solvent.

7. A modified hydroxytyrosol, characterized in that, The modified hydroxytyrosol is prepared by the method according to any one of claims 1-6, wherein the modified hydroxytyrosol comprises a volatile component and a compound having the structure shown in the following formula: 。 8. The modified hydroxytyrosol according to claim 7, characterized in that, The volatile components include at least one of aromatic compounds, pyranones, sugars, and urea.

9. The use of modified hydroxytyrosol prepared by the method of any one of claims 1-6 or the modified hydroxytyrosol according to any one of claims 7-8 in the preparation of cosmetics.

10. A cosmetic product, characterized in that, The modified hydroxytyrosol prepared by the method of any one of claims 1-6 or the modified hydroxytyrosol according to any one of claims 7-8.

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