Hydroxytyrosol inclusion with stabilizing effect and preparation method thereof

Abstract

The invention discloses a hydroxytyrosol inclusion with a stabilizing effect and a preparation method thereof. The raw materials of the hydroxytyrosol inclusion of the invention include hydroxytyrosol, solid lipids, an emulsifier and water. The invention adopts a water-in-oil method to prepare the hydroxytyrosol inclusion, combines supramolecular technology with solid nanolipid technology, and dissolves hydroxytyrosol in solid fats and oils by designing and screening solid lipids, and rapidly emulsifies and rapidly cools to prepare the hydroxytyrosol inclusion with a stabilizing effect. The hydroxytyrosol inclusion prepared by the invention has good stability, and long-term storage will not cause precipitation and discoloration of active substances. At the same time, the application scenarios are wider and can better meet the system types of most daily chemical products.

Description

Technical Field

[0001] The invention belongs to the technical field of daily chemical industry, and particularly relates to a hydroxytyrosol inclusion body with stabilizing effect and a preparation method thereof. Background Art

[0002] Whitening cosmetics are one of the important cosmetic categories. With the improvement of economic level, consumers' demand for whitening cosmetics is increasing. Therefore, the development and utilization of new whitening agent raw materials have important practical significance for the research and development of whitening cosmetics.

[0003] Hydroxytyrosol (3,4-Dihydroxyphenylethanol, Figure 1 ) is the main active ingredient of olive leaves and is a new type of whitening agent molecule. Studies have shown that the production process of melanin starts with the oxidation of L-tyrosine to dopaquinone by tyrosinase, followed by a series of enzyme-catalyzed oxidations to produce two types of melanin, eumelanin (brown-black) and pheomelanin (red-yellow). Tyrosinase is the only key enzyme currently known to participate in melanin synthesis in organisms. Hydroxytyrosol has tyrosinase inhibitory activity and can intervene in multiple stages of melanin formation to achieve a whitening effect. At the same time, hydroxytyrosol has multiple effects such as antioxidant and anti-aging, and is a very potential natural active molecule. However, like many polyphenol molecules, hydroxytyrosol is extremely sensitive to temperature and pH and is prone to discoloration, which greatly hinders its use in cosmetics.

[0004] Currently, encapsulation technology is commonly used to solve the problem of discoloration of active ingredients, such as nanoemulsions, microemulsions, liposomes, solid lipid nanoparticles, etc. However, these technologies are generally targeted at oil-soluble active ingredients, and cannot effectively deal with the discoloration problem of water-soluble active ingredients. Due to its small molecular weight and many hydrophilic groups, hydroxytyrosol is highly soluble in water and can be mixed with water in any proportion, but it is insoluble in common oil phases. The development of stabilized formulation technology for hydroxytyrosol is very challenging.

[0005] As a whitening agent, hydroxytyrosol is easy to change color, which greatly limits its application in product formulations. Patent document CN114099357A proposes an oil-in-polyol nanoemulsion that can improve the stability of hydroxytyrosol and inhibit its oxidation. However, the application scenarios of oil-in-polyol nanoemulsions in daily chemical products are very limited. Most daily chemical products use water as the continuous phase, and the creams and emulsions of manufacturers are all oil-in-water emulsions. Therefore, the development of hydroxytyrosol encapsulation technology in the water phase has higher technical value and practical significance. Summary of the invention

[0006] In view of the deficiencies of the prior art, the present invention provides a hydroxytyrosol inclusion with a stabilizing effect and a preparation method thereof. The present invention combines supramolecular technology with solid nanolipid technology, designs and screens solid lipids, utilizes the complexing properties between solid lipids and hydroxytyrosol, dissolves hydroxytyrosol in solid fats and oils, and rapidly emulsifies and rapidly cools to prepare a hydroxytyrosol inclusion with a stabilizing effect. The inclusion containing hydroxytyrosol proposed by the present invention has good stability, and long-term storage will not cause the active substance to precipitate or discolor.

[0007] The technical solution of the present invention is as follows:

[0008] A hydroxytyrosol inclusion with a stabilizing effect. The raw materials of the hydroxytyrosol inclusion include hydroxytyrosol, solid lipid, emulsifier and water.

[0009] Furthermore, the solid lipid is a fatty acid, and the fatty acid includes one or more of lauric acid, myristic acid, palmitic acid, eicosanoic acid, and behenic acid.

[0010] Furthermore, the emulsifier is a fatty alcohol polyether product corresponding to solid lipids; the number of polyether units in the fatty alcohol polyether product is 5-40; the emulsifier includes one or more of lauryl alcohol polyether, myristyl alcohol polyether, cetyl alcohol polyether, stearyl alcohol polyether, eicosanol polyether, and behenyl alcohol polyether.

[0011] Furthermore, the mass ratio of the solid lipid to hydroxytyrosol is >2:1, preferably 3-5:1.

[0012] Furthermore, the mass ratio of the emulsifier to the solid lipid is 1:5-10, preferably 1:5-8.

[0013] Furthermore, the preparation method comprises the following steps:

[0014] (1) mixing hydroxytyrosol and solid lipids and heating them to obtain a liquid oil phase, and then adding an emulsifier and stirring to dissolve them to obtain a phase a;

[0015] (2) using water as phase b, homogenizing phase b in a water bath, and gradually adding phase a prepared in step (1) during the homogenization process to obtain an emulsion;

[0016] (3) pouring the emulsion of step (2) into water, dissolving by stirring and cooling, thereby forming a solid lipid nanoemulsion, i.e., an oil-in-water type hydroxytyrosol inclusion body.

[0017] Furthermore, in step (1), the heating temperature is 60-90° C. and the heating time is 15-25 min; the stirring is mechanical stirring with a rotation speed of 200-300 rpm and a stirring time of 3-5 min.

[0018] Furthermore, in step (2), the mass ratio of water to solid lipid is 4-9:1, preferably 4-8:1.

[0019] Furthermore, in step (2), the temperature of the water bath is 80-90° C.; the homogenization time is 3-5 min, and the rotation speed is 10000-18000 rpm.

[0020] Furthermore, in step (3), the mass ratio of the emulsion to water is 1:3-10; the cooling is specifically pouring the homogenized emulsion into water and rapidly cooling it to 20-30°C by rapid stirring; the stirring is mechanical stirring, the stirring speed is 300-400 rpm, and the time is 10-15 min.

[0021] The beneficial technical effects of the present invention are:

[0022] The present invention combines supramolecular technology with solid nanolipid technology, and through designing and screening solid lipids, utilizing the complexation properties between solid lipids and hydroxytyrosol, a relatively stable supramolecular structure is realized through secondary bonds such as hydrogen bonds, so that hydroxytyrosol is dissolved in solid fats and oils; the emulsifier corresponding to the solid fats and oils is selected for the purpose of constructing an environment conducive to the formation of supramolecular structures. Hydroxytyrosol inclusions with stabilization effects are prepared by rapid emulsification and rapid cooling, and the core lies in encapsulating hydroxytyrosol in solid oil droplets of nanoemulsions, isolating it from the external environment, thereby avoiding oxidation of hydroxytyrosol.

[0023] The hydroxytyrosol inclusions provided by the present invention have good stability, and long-term storage will not cause the active substance to precipitate or discolor. The hydroxytyrosol inclusions prepared by the present invention have wider application scenarios and can better meet the system types of most daily chemical products. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is the structural formula of hydroxytyrosol.

[0025] Figure 2 This is the standard curve of hydroxytyrosol.

[0026] Figure 3 This is a curve diagram showing the change of hydroxytyrosol retention rate over time of the hydroxytyrosol inclusions prepared in Example 1 of the present invention under storage conditions of 50°C.

[0027] Figure 4 This is a comparison chart of the hydroxytyrosol inclusions prepared in Example 1 of the present invention stored at a constant temperature of 50° C. for 14 days.

[0028] In the figure: 1, the initial state of the hydroxytyrosol inclusions prepared in Example 1; 2, the state of the hydroxytyrosol inclusions prepared in Example 1 after being stored at a constant temperature of 50° C. for 14 days.

[0029] Figure 5 The hydroxytyrosol inclusions prepared in Examples 1-12.

[0030] In the figure: 1, hydroxytyrosol inclusions prepared in Example 1; 2, hydroxytyrosol inclusions prepared in Example 2; 3, hydroxytyrosol inclusions prepared in Example 3; 4, hydroxytyrosol inclusions prepared in Example 4; 5, hydroxytyrosol inclusions prepared in Example 5; 6, hydroxytyrosol inclusions prepared in Example 6; 7, hydroxytyrosol inclusions prepared in Example 7; 8, hydroxytyrosol inclusions prepared in Example 8; 9, hydroxytyrosol inclusions prepared in Example 9; 10, hydroxytyrosol inclusions prepared in Example 10; 11, hydroxytyrosol inclusions prepared in Example 11; 12, hydroxytyrosol inclusions prepared in Example 12.

[0031] Figure 6 In the hydroxytyrosol inclusion bodies prepared in Example 1 of the present invention, behenic acid and hydroxytyrosol form a supramolecular system.

[0032] Figure 7 This is the emulsion prepared in Comparative Example 1 of the present invention.

[0033] Figure 8 This is the emulsion prepared in Comparative Examples 2-4 of the present invention.

[0034] Fig. 9 This is the emulsion prepared in Comparative Example 5 of the present invention. DETAILED DESCRIPTION

[0035] The present invention is described in detail below in conjunction with the accompanying drawings and embodiments.

[0036] Embodiment 1:

[0037] A hydroxytyrosol inclusion with a stabilizing effect, the preparation method of which comprises the following steps:

[0038] Take a 100ml beaker, add 4g hydroxytyrosol and 20g behenic acid, heat at 80℃ for 20min to obtain a liquid oil phase, then add 4g behenyl alcohol polyether-20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; take 80g water as phase b, homogenize phase b at 18000rpm for 3min under 80℃ water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the above emulsion and pour it into 400g water, stir at 400rpm for 10min, completely dissolve it and cool it to 20℃ to form a solid lipid nanoemulsion, that is, the active substance hydroxytyrosol inclusion body proposed in the present invention is obtained.

[0039] Embodiment 2:

[0040] A hydroxytyrosol inclusion with a stabilizing effect, the preparation method of which comprises the following steps:

[0041] Take a 100ml beaker, add 4g hydroxytyrosol and 20g palmitic acid, heat at 80℃ for 20min to obtain a liquid oil phase, then add 4g hexadecanol polyether-20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; take 80g water as phase b, homogenize phase b at 18000rpm for 3min under 80℃ water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the emulsion and pour it into 400g water, stir at 400rpm for 10min, completely dissolve it and cool it to 20℃ to form a solid lipid nanoemulsion, that is, the active hydroxytyrosol inclusion body proposed in the present invention is obtained.

[0042] Embodiment 3:

[0043] A hydroxytyrosol inclusion with a stabilizing effect, the preparation method of which comprises the following steps:

[0044] Take a 100ml beaker, add 4g hydroxytyrosol, 10g palmitic acid and 10g behenic acid, heat at 60℃ for 25min to obtain a liquid oil phase, then add 2g behenyl alcohol polyether-20 and mix with 2g hexadecanol polyether-20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; take 80g water as phase b, homogenize phase b at 10000rpm for 5min under 80℃ water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the above emulsion and pour it into 400g water, stir at 300rpm for 15min, completely dissolve it and cool it to 20℃ to form a solid lipid nanoemulsion, that is, the active substance hydroxytyrosol inclusion body proposed in the present invention is obtained.

[0045] Embodiment 4:

[0046] A hydroxytyrosol inclusion with a stabilizing effect, the preparation method of which comprises the following steps:

[0047] Take a 100ml beaker, add 4g hydroxytyrosol and 20g behenic acid, heat at 80℃ for 20min to obtain a liquid oil phase, then add 3.3g behenyl alcohol polyether-20 as an emulsifier, stir at 300rpm for 3min, and completely dissolve it to a clear and transparent state to obtain phase a; take 80g water as phase b, homogenize phase b at 18000rpm for 3min under 80℃ water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 2L ml beaker and add 1000g water, homogenize the above emulsion and pour it into 400g water, stir at 400rpm for 10min, completely dissolve it and cool it to 20℃, to form a solid lipid nanoemulsion, that is, the active substance hydroxytyrosol inclusion body proposed in the present invention is obtained.

[0048] Embodiment 5:

[0049] A hydroxytyrosol inclusion with a stabilizing effect, the preparation method of which comprises the following steps:

[0050] Take a 100ml beaker, add 4g hydroxytyrosol and 20g behenic acid, heat at 90℃ for 15min to obtain a liquid oil phase, then add 2.5g behenyl alcohol polyether-20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; take 80g water as phase b, homogenize phase b at 18000rpm for 3min under 80℃ water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the above emulsion and pour it into 400g water, stir at 400rpm for 10min, completely dissolve it and cool it to 20℃ to form a solid lipid nanoemulsion, that is, the active hydroxytyrosol inclusion body proposed in the present invention is obtained.

[0051] Embodiment 6:

[0052] A hydroxytyrosol inclusion with a stabilizing effect, the preparation method of which comprises the following steps:

[0053] Take a 100ml beaker, add 4g hydroxytyrosol and 20g behenic acid, heat at 80℃ for 20min to obtain a liquid oil phase, then add 2g behenyl alcohol polyether-20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; take 80g water as phase b, homogenize phase b at 18000rpm for 3min under 90℃ water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the above emulsion and pour it into 400g water, stir at 400rpm for 10min, completely dissolve it and cool it to 20℃, to form a solid lipid nanoemulsion, that is, the active substance hydroxytyrosol inclusion body proposed in the present invention is obtained.

[0054] Embodiment 7:

[0055] A hydroxytyrosol inclusion with a stabilizing effect, the preparation method of which comprises the following steps:

[0056] Take a 100ml beaker, add 4g hydroxytyrosol and 20g behenic acid, heat at 80℃ for 20min to obtain a liquid oil phase, then add 4g behenyl alcohol polyether-20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; take 120g water as phase b, homogenize phase b at 18000rpm for 3min under 80℃ water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the above emulsion and pour it into 400g water, stir at 400rpm for 10min, completely dissolve it and cool it to 20℃ to form a solid lipid nanoemulsion, that is, the active hydroxytyrosol inclusion body proposed in the present invention is obtained.

[0057] Embodiment 8:

[0058] A hydroxytyrosol inclusion with a stabilizing effect, the preparation method of which comprises the following steps:

[0059] Take a 100ml beaker, add 4g hydroxytyrosol and 20g behenic acid, heat at 80℃ for 20min to obtain a liquid oil phase, then add 4g behenyl alcohol polyether-20 as an emulsifier, stir at 200rpm for 5min to completely dissolve it to a clear and transparent state to obtain phase a; take 160g water as phase b, homogenize phase b at 18000rpm for 3min under 80℃ water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the above emulsion and pour it into 1200g water, cool it to 20℃, stir at 400rpm for 10min to completely dissolve it, and form a solid lipid nanoemulsion, that is, the active hydroxytyrosol inclusion body proposed in the present invention is obtained.

[0060] Embodiment 9:

[0061] A hydroxytyrosol inclusion with a stabilizing effect, the preparation method of which comprises the following steps:

[0062] Take a 100ml beaker, add 4g hydroxytyrosol and 20g behenic acid, heat at 80℃ for 20min to obtain a liquid oil phase, then add 4g behenyl alcohol polyether-20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; take 180g water as phase b, homogenize phase b at 18000rpm for 3min under 80℃ water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 1000ml beaker and add 400g water, homogenize the above emulsion and pour it into 1200g water, stir at 400rpm for 10min, completely dissolve it and cool it to 20℃ to form a solid lipid nanoemulsion, that is, the active substance hydroxytyrosol inclusion body proposed in the present invention is obtained.

[0063] Embodiment 10:

[0064] A hydroxytyrosol inclusion with a stabilizing effect, the preparation method of which comprises the following steps:

[0065] Take a 100ml beaker, add 5g hydroxytyrosol and 20g behenic acid, heat at 80℃ for 20min to obtain a liquid oil phase, then add 4g behenyl alcohol polyether-20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; take 80g water as phase b, homogenize phase b at 18000rpm for 3min under 80℃ water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the above emulsion and pour it into 400g water, stir at 400rpm for 10min, completely dissolve it and cool it to 20℃ to form a solid lipid nanoemulsion, that is, the active substance hydroxytyrosol inclusion body proposed in the present invention is obtained.

[0066] Embodiment 11:

[0067] A hydroxytyrosol inclusion with a stabilizing effect, the preparation method of which comprises the following steps:

[0068] Take a 100ml beaker, add 6.7g hydroxytyrosol and 20g behenic acid, heat at 80℃ for 20min to obtain a liquid oil phase, then add 4g behenyl alcohol polyether-20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; take 80g water as phase b, homogenize phase b at 18000rpm for 3min under 80℃ water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the above emulsion and pour it into 400g water, stir at 400rpm for 10min, completely dissolve it and cool it to 20℃ to form a solid lipid nanoemulsion, that is, the active substance hydroxytyrosol inclusion body proposed in the present invention is obtained.

[0069] Embodiment 12:

[0070] A hydroxytyrosol inclusion with a stabilizing effect, the preparation method of which comprises the following steps:

[0071] Take a 100ml beaker, add 10g hydroxytyrosol and 20g behenic acid, heat at 80℃ for 20min to obtain a liquid oil phase, then add 4g behenyl alcohol polyether-20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; take 80g water as phase b, homogenize phase b at 18000rpm for 3min under 80℃ water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the above emulsion and pour it into 400g water, stir at 400rpm for 10min, completely dissolve it and cool it to 20℃ to form a solid lipid nanoemulsion, that is, the active hydroxytyrosol inclusion body proposed in the present invention is obtained.

[0072] Comparative Example 1:

[0073] The solid lipid in Example 1 was replaced with behenyl alcohol, and the rest remained unchanged, as Comparative Example 1. Specifically,

[0074] Take a 100 ml beaker, add 4 g of hydroxytyrosol and 20 g of behenyl alcohol, heat at 80°C for 20 min, then add 4 g of behenyl alcohol polyether-20 as an emulsifier, stir at 200 rpm for 5 min, and it is found that hydroxytyrosol cannot be dissolved in the oil phase.

[0075] Comparative Example 2:

[0076] The solid lipid in Example 1 was replaced with lauric acid, and the rest remained unchanged, as Comparative Example 2. Specifically, take a 100ml beaker, add 4g hydroxytyrosol and 20g lauric acid, heat at 80℃ for 20min, then add 4g behenyl alcohol polyether-20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; 80g water was used as phase b, and phase b was homogenized at 18000rpm for 3min under 80℃ water bath conditions, and phase a was gradually added during the homogenization process to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the above emulsion and pour it into 400g water, stir at 400rpm for 10min, cool to 20℃, the resulting emulsion has solid precipitation, and the solid agglomerates and separates when standing.

[0077] Comparative Example 3:

[0078] Replace the emulsifier in Example 1 with Tween 20, and keep the rest unchanged, as Comparative Example 3. Specifically, take a 100ml beaker, add 4g hydroxytyrosol and 20g behenic acid, heat at 80℃ for 20min, then add 4g Tween 20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; take 80g water as phase b, homogenize phase b at 18000rpm for 3min in an 80℃ water bath, gradually add phase a during the homogenization process to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the above emulsion and pour it into 400g water, stir at 400rpm for 10min, cool to 20℃, the resulting emulsion has solid precipitation, and the solid agglomerates and separates when left to stand.

[0079] Comparative Example 4:

[0080] The amount of the emulsifier behenyl alcohol polyether-20 in Example 1 was replaced with 1g, and the rest remained unchanged, as comparative example 4. Specifically, take a 100ml beaker, add 4g hydroxytyrosol and 20g behenic acid, heat at 80℃ for 20min, then add 1g behenyl alcohol polyether-20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; 80g water is used as phase b, and phase b is homogenized at 18000rpm for 3min under 80℃ water bath conditions, and phase a is gradually added during the homogenization process to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the above emulsion and pour it into 400g water, stir at 400rpm for 10min, cool to 20℃, the resulting emulsion has solid precipitation, and the solid agglomerates and separates when left to stand.

[0081] Comparative Example 5:

[0082] The amount of water in Example 1 was replaced with 50g, and the rest remained unchanged, as Comparative Example 5. Specifically, take a 100ml beaker, add 4g hydroxytyrosol and 20g behenic acid, heat at 80℃ for 20min, obtain a liquid oil phase, then add 4g behenyl alcohol polyether-20 as an emulsifier, stir at 200rpm for 5min, and completely dissolve it to a clear and transparent state to obtain phase a; take 50g water as phase b, homogenize phase b at 18000rpm for 3min under 80℃ water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 1L beaker and add 400g water, homogenize the above emulsion and pour it into 400g water, stir at 400rpm for 10min, cool to 20℃, and the obtained emulsion is stored at a constant temperature of 50℃ for 7 days, and the color change is obvious.

[0083] Comparative Example 6:

[0084] The "after homogenizing the emulsion, pour it into 400g of water" in Example 1 was replaced with "after homogenizing the emulsion, pour it into 200g of water", and the rest remained unchanged, as Comparative Example 6. Specifically, a 100ml beaker was taken, 4g of hydroxytyrosol and 20g of behenic acid were added, and heated at 80°C for 20min to obtain a liquid oil phase, and then 4g of behenyl alcohol polyether-20 was added as an emulsifier, and stirred at 200rpm for 5min to completely dissolve it into a clear and transparent state to obtain phase a; 80g of water was used as phase b, and phase b was homogenized at 18000rpm for 3min under 80°C water bath conditions, and phase a was gradually added during the homogenization process to obtain an emulsion. Prepare another 1L beaker and add 200g of water. Pour the above emulsion into 200g of water after homogenization, stir at 400rpm for 10min, and cool to 20°C. The lipid in the obtained emulsion is not cooled in time, and hydroxytyrosol dissolves in the aqueous phase, resulting in low encapsulation efficiency, obvious discoloration, and poor stabilization effect.

[0085] Comparative Example 7:

[0086] Replace "stirring at 400 rpm for 10 min" in Example 1 with "stirring at 100 rpm for 5 min", and keep the rest unchanged as Comparative Example 7. Specifically, take a 100 ml beaker, add 4 g hydroxytyrosol and 20 g behenic acid, heat at 80 ° C for 20 min, obtain a liquid oil phase, then add 4 g behenyl alcohol polyether-20 as an emulsifier, stir at 200 rpm for 5 min, and completely dissolve it to a clear and transparent state to obtain phase a; take 80 g water as phase b, homogenize phase b at 18000 rpm for 3 min under 80 ° C water bath conditions, gradually add phase a during homogenization to obtain an emulsion. Prepare another 1L beaker and add 400 g water, homogenize the above emulsion and pour it into 400 g water, stir at 400 rpm for 10 min, and cool to 20 ° C. The lipids in the obtained emulsion are not cooled in time, and hydroxytyrosol dissolves in the water phase, resulting in low encapsulation efficiency, obvious discoloration, and poor stabilization effect.

[0087] Test example:

[0088] (1) Thermal stability of hydroxytyrosol:

[0089] The hydroxytyrosol inclusions prepared in Example 1 were taken to detect the change of hydroxytyrosol retention rate over time. The specific detection method was as follows: ① 0.3, 0.6, 1.0, 1.3, and 1.5 mg / ml hydroxytyrosol methanol solutions were prepared respectively, and the absorbance values ​​were measured with an ultraviolet spectrophotometer to obtain a hydroxytyrosol standard curve, such as Figure 2As shown; ② The prepared hydroxytyrosol inclusions were stored at 50°C, a small amount of hydroxytyrosol inclusions were taken at regular intervals every day, diluted with methanol by a certain multiple (so that the absorbance falls within the range of 0.2-0.6), and the absorbance value was measured with an ultraviolet spectrophotometer. The hydroxytyrosol content in the hydroxytyrosol inclusions was obtained by comparing the hydroxytyrosol standard curve and multiplying by the dilution multiple; ③ The hydroxytyrosol retention rate was calculated: the hydroxytyrosol retention rate on the nth day (%) = the hydroxytyrosol content on the nth day / the hydroxytyrosol content on the 0th day × 100% (the hydroxytyrosol content on the 0th day is the initial state of the hydroxytyrosol inclusions obtained in Example 1). The results are shown in FIG. Figure 3 , 4 As shown. From the hydroxytyrosol retention rate curve over time, it can be seen that the hydroxytyrosol retention rate of the solid nano lipid emulsion prepared in Example 1 decreases slowly over time under the storage condition of 50°C. At 25 days, the hydroxytyrosol retention rate is still 90%. At the same time, from the comparison between the emulsion kept at 50°C for 14 days and the initial state, it can be seen that there is no precipitation of the active substance hydroxytyrosol in the emulsion, and no discoloration, stratification, etc.

[0090] (2) Relationship between hydroxytyrosol solubility and oil phase type:

[0091] Take the emulsion prepared in Comparative Example 1 and observe its dissolution. The results are as follows: Figure 7 As shown in the figure, it can be seen that ordinary oil phases such as fatty alcohols, triglycerides, mineral waxes, higher fatty acid higher esters and other solid lipids cannot dissolve hydroxytyrosol. The type of oil phase is crucial to the formation of the emulsion proposed by the present invention. The reason is that the carbonyl group of fatty acids can act as a hydrogen bond acceptor, and the hydroxyl group of hydroxytyrosol can act as a hydrogen bond donor, and the two form a supramolecular system, such as Figure 6 After the formation of supramolecules, the hydrophilic groups are shielded, the number is reduced, and the oil solubility is increased.

[0092] (3) Relationship between hydroxytyrosol solubility and oil phase / emulsifier compatibility:

[0093] Take the emulsion prepared in Comparative Example 2 and observe its dissolution. The results are as follows: Figure 8 As shown in the figure, it can be seen that the emulsion prepared in Comparative Example 2 has solid precipitation, and the solid agglomerates and delaminates when left to stand, indicating that the compatibility of the oil phase and the emulsifier is crucial to the formation of the hydroxytyrosol inclusion body proposed in the present invention.

[0094] (4) Relationship between hydroxytyrosol solubility and emulsifier type:

[0095] Take the emulsion prepared in Comparative Example 3 and observe its dissolution. The results are as follows: Figure 8As shown in the figure, it can be seen that the emulsion prepared in Comparative Example 3 has solid precipitation, and the solid agglomerates and separates when left to stand, indicating that the type of emulsifier is crucial to the formation of the hydroxytyrosol inclusion body proposed in the present invention.

[0096] (5) Relationship between hydroxytyrosol solubility and emulsifier dosage:

[0097] Take the emulsion prepared in Comparative Example 4 and observe its dissolution. The results are as follows: Figure 8 As shown in the figure, it can be seen that the emulsion prepared in Comparative Example 4 has solid precipitation, and the solid agglomerates and separates when left to stand, indicating that the selection of the amount of emulsifier is crucial to the formation of the hydroxytyrosol inclusion body proposed in the present invention.

[0098] (6) Relationship between hydroxytyrosol stability and oil-water ratio:

[0099] Take the emulsion prepared in Comparative Example 5 and observe its dissolution. The results are as follows: Fig. 9 As shown in the figure, the emulsion prepared in Comparative Example 5 changes color significantly after being stored at a constant temperature of 50°C for 7 days, indicating that the selection of the oil-water ratio is crucial to the stability of the hydroxytyrosol inclusion body proposed in the present invention, and the emulsion prepared by the oil-water ratio of Comparative Example 5 has a poor encapsulation effect on hydroxytyrosol.

[0100] The above is only a preferred embodiment of the present invention, and the present invention is not limited to the above embodiments. It is understood that other improvements and changes directly derived or associated by those skilled in the art without departing from the spirit and concept of the present invention should be considered to be included in the protection scope of the present invention.

Claims

1. A hydroxytyrosol inclusion with a stabilizing effect, characterized in that: The raw materials of the hydroxytyrosol inclusion body include hydroxytyrosol, solid lipid, emulsifier and water; The solid lipid is a fatty acid, and the fatty acid includes one or more of palmitic acid, eicosanoic acid, and behenic acid; The emulsifier is a fatty alcohol polyether product corresponding to the solid lipid; the number of polyether units in the fatty alcohol polyether product is 5-40; the emulsifier includes one or more of cetyl alcohol polyether, eicosanol polyether, and behenyl alcohol polyether; The mass ratio of the solid lipid to hydroxytyrosol is >2:1; The mass ratio of the emulsifier to the solid lipid is 1:5-10; The preparation method of the hydroxytyrosol inclusion body comprises the following steps: (1) mixing hydroxytyrosol and solid lipids and heating them to obtain a liquid oil phase, and then adding an emulsifier and stirring to dissolve them to obtain a phase a; (2) using water as phase b, homogenizing phase b in a water bath, and gradually adding phase a prepared in step (1) during the homogenization process to obtain an emulsion; (3) pouring the emulsion of step (2) into water, dissolving by stirring and cooling to form a solid lipid nanoemulsion, i.e., hydroxytyrosol inclusion bodies; In step (2), the mass ratio of water to solid lipid is 4-9:1; In step (3), the mass ratio of the emulsion to water is 1:3-10; the cooling is specifically pouring the homogenized emulsion into water and rapidly cooling it to 20-30°C by rapid stirring; the stirring is mechanical stirring at a speed of 300-400 rpm for 10-15 min.

2. The hydroxytyrosol inclusion body according to claim 1, characterized in that The mass ratio of the solid lipid to hydroxytyrosol is 3-5:

1.

3. The hydroxytyrosol inclusion body according to claim 1, characterized in that The mass ratio of the emulsifier to the solid lipid is 1:5-8.

4. The hydroxytyrosol inclusion body according to claim 1, characterized in that: In step (1), the heating temperature is 60-90° C. and the heating time is 15-25 min; the stirring is mechanical stirring at a rotation speed of 200-300 rpm and the stirring time is 3-5 min.

5. The hydroxytyrosol inclusion body according to claim 1, characterized in that: In step (2), the mass ratio of water to solid lipid is 4-8:

1.

6. The hydroxytyrosol inclusion body according to claim 1, characterized in that: In step (2), the temperature of the water bath is 80-90° C.; the homogenization time is 3-5 min, and the rotation speed is 10000-18000 rpm.

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