A moisturizing and soothing enzymatic noni seed oil, and a preparation method and application thereof
By isolating and immobilizing lipases from germinated noni seeds to hydrolyze noni seed oil, the problems of high production cost and limited efficacy of noni seed oil have been solved, resulting in a highly active and multifunctional hydrolyzed noni seed oil that can be used in cosmetics.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- COSMAX CHINA INC
- Filing Date
- 2024-10-23
- Publication Date
- 2026-06-12
AI Technical Summary
The production cost of existing noni seed oil is high and its efficacy is limited, making it difficult to widely use in cosmetics.
By isolating and immobilizing lipase from germinated noni seeds, enzymatically hydrolyzing cold-pressed noni seed oil, and recycling the highly active enzymatically hydrolyzed oil, enzymatically hydrolyzed noni seed oil with moisturizing and soothing effects can be prepared.
The prepared enzymatically hydrolyzed noni seed oil has higher active ingredients and efficacy, and exhibits significant moisturizing, soothing and emulsifying abilities when applied to cosmetics.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of cosmetics, and more particularly to the A61K8 field. Specifically, it relates to a moisturizing and soothing enzymatically hydrolyzed noni seed oil, its preparation method, and its application. Background Technology
[0002] Noni fruit (Morinda citrifolia) is the fruit of the seashore tree Morinda. Used as food and medicine in Polynesia for thousands of years, the medicinal uses of the fruit, juice, flowers, leaves, bark, and roots are well-known locally; however, little is known about the potential health benefits of oil extracted from the seeds.
[0003] Noni seeds were long considered a byproduct of the noni juice industry until a process for extracting noni seed oil was developed. A single noni fruit contains 200-250 seeds, but it takes thousands of seeds to produce just 0.024 liters of noni seed oil, making it a valuable resource. While small-scale production of noni seed oil is prohibitively expensive, it becomes economically viable when linked to large-scale noni juice production.
[0004] The existing technologies CN103550095A, CN106377453A and CN106214579A disclose the application of noni fruit in cosmetics, all of which use noni fruit extract, and the skin care effects are all focused on whitening. The effects are relatively simple, and the production cost is high, which is not conducive to market promotion and practical application. Summary of the Invention
[0005] To address the aforementioned technical problems, the first aspect of this invention provides a method for preparing enzymatically hydrolyzed noni seed oil (Morinda citrifolia) with moisturizing and soothing effects, comprising the following steps:
[0006] S1. Prepare fresh noni seeds, wash them, and then germinate them in water.
[0007] S2. Germinated noni seeds from 1 to 7 days were taken out, ground, centrifuged, and fractionated to separate the oil bodies from the 1 to 7-day-old noni seeds, obtain the immobilized lipase in the oil bodies, and detect the activity of the immobilized lipase.
[0008] S3. Select the immobilized lipase with the highest activity for enzymatic hydrolysis of cold-pressed noni seed oil;
[0009] S4. After enzymatic hydrolysis, the hydrolyzed noni seed oil containing lipase is separated and stored for recycling in the next oil preparation.
[0010] Furthermore, in step S1, the noni seeds are from noni fruits originating in Hainan.
[0011] Furthermore, in step S2, grinding and centrifugation are both carried out at 3-7°C. 10-25g of germinated noni seeds are ground into a homogenate in a mortar, and the pH is adjusted to 5-7. The homogenate is filtered through four layers of cheesecloth, and the filtrate is centrifuged at 1000g for 10min to obtain crude cell extract (supernatant). It is then centrifuged at 3500g for 20min and fractionated. The crude supernatant is taken and mixed with an equal volume of ether four times to extract neutral oil. It is then mixed with ether again, washed with Na2SO4 aqueous solution to remove peptides, and concentrated until an oil body is obtained. The remaining ether is evaporated under nitrogen, and the composition of the obtained oil is analyzed by thin-layer chromatography. Then, it is used as a substrate to qualitatively test the enzyme activity of immobilized lipase.
[0012] Furthermore, in step S3, the pH of the oil body system is first adjusted to 7-10, the oil body is placed in a reaction vessel, the temperature is controlled at 35-43℃, and 33 mmol / L emulsified gum arabic triolein, 12.8 mmol / L deoxycholate, and 10.7 mmol / L NaCl are added; then the lipase activity is measured using p-nitrophenyl palmitate; based on the measurement results, the immobilized lipase with the highest activity is selected and added to cold-pressed fresh noni seed oil for enzymatic hydrolysis for 1-7 days; wherein the emulsified gum arabic triolein, deoxycholate, and NaCl are in equal volumes.
[0013] Furthermore, in step S4, after enzymatic hydrolysis is completed, the hydrolyzed noni seed oil containing the most active immobilized lipase is separated and preserved by centrifugation, which can be used for the next enzymatic hydrolysis, thus achieving recycling.
[0014] More preferably, the hydroponic germination time of the noni seeds in step S1 is 4-6 days.
[0015] More preferably, the S2 grinding and centrifugation are both carried out at 4-6℃. 15-20g of germinated noni seeds are ground into a homogenate in a mortar, and the pH is adjusted to 6. The homogenate is filtered through four layers of cheesecloth, and the filtrate is centrifuged at 1000g for 10min to obtain crude cell extract (supernatant). It is then centrifuged at 3500g for 20min and fractionated. The crude supernatant is taken and mixed with an equal volume of ether four times to extract neutral oil. The ether phase is mixed and washed with Na2SO4 aqueous solution to remove peptides, and concentrated until an oil body is obtained. The remaining ether is evaporated under nitrogen, and the composition of the obtained oil is analyzed by thin-layer chromatography. Then, it is used as a substrate to qualitatively test the enzyme activity of immobilized lipase.
[0016] More preferably, in step S3, the pH of the oil body system is first adjusted to 8-9, the oil body is placed in a reaction vessel, the temperature is controlled at 36-40℃, and 33 mmol / L emulsified gum arabic triolein, 12.8 mmol / L deoxycholate and 10.7 mmol / L NaCl are added; the purity of the emulsified gum arabic triolein used in the substrate emulsion is determined by thin-layer chromatography; then the lipase activity is determined by p-nitrophenyl palmitate; based on the measurement results, the immobilized lipase with the highest activity is selected and added to cold-pressed fresh noni seed oil for enzymatic hydrolysis for 3-5 days.
[0017] The second invention provides an enzymatically hydrolyzed noni seed oil prepared by the above preparation method.
[0018] The third invention provides the application of enzymatically hydrolyzed noni seed oil in cosmetics.
[0019] Preferably, the cosmetic product includes one or more of the following: serum, emulsion, cream, lotion, and mask.
[0020] Preferably, the enzymatically hydrolyzed noni seed oil is added to cosmetics at a mass percentage of 0.5-10%.
[0021] Beneficial effects
[0022] (i) This invention separates the oil body from germinated noni seeds, obtains the oil body component of lipase embedded in the natural noni seed oil body, and uses it as an immobilized lipase for enzymatic hydrolysis of cold-pressed noni seed oil.
[0023] (ii) The present invention detects the activity of immobilized lipase in the oil bodies isolated from noni seeds that have been germinated for 1-7 days, and selects the best oil bodies from the germinated seeds for enzymatic hydrolysis and cold pressing of noni seed oil.
[0024] (III) After the enzymatic hydrolysis is completed, the highly active immobilized lipase oleosomes and the hydrolyzed noni seed oil are separated and reused as the source of hydrolysate for the next reaction.
[0025] (iv) The enzymatically hydrolyzed Noni seed oil obtained by the process described in this invention has moisturizing and soothing effects, and its emulsifying ability is stronger when applied in formulations.
[0026] (v) This invention utilizes fixed lipase oleosomes extracted from germinated noni seeds to enzymatically hydrolyze cold-pressed fresh noni seed oil. Compared with unhydrolyzed noni seed oil, the hydrolyzed noni seed oil has higher active ingredients and better efficacy. Attached Figure Description
[0027] Figure 1 The graph shows the results of the emulsification performance test.
[0028] Figure 2 This is a diagram showing the test results for oil droplet morphology. Detailed Implementation
[0029] Example 1
[0030] The first aspect of this example provides a method for preparing enzymatically hydrolyzed noni seed oil with moisturizing and soothing effects, which consists of the following steps:
[0031] S1. Prepare fresh noni seeds, wash them, and then germinate them in water.
[0032] S2. Germinated noni seeds from 1 to 7 days were taken out, ground, centrifuged, and fractionated to separate the oil bodies from the 1 to 7-day-old noni seeds, obtain the immobilized lipase in the oil bodies, and detect the activity of the immobilized lipase.
[0033] S3. Select the immobilized lipase with the highest activity for enzymatic hydrolysis of cold-pressed noni seed oil;
[0034] S4. After enzymatic hydrolysis, the hydrolyzed noni seed oil containing lipase is separated and stored for recycling in the next oil preparation.
[0035] In step S1, the noni seeds are from noni fruits originating in Hainan, and the hydroponic germination time is 5 days.
[0036] The S2 grinding and centrifugation were both carried out at 5°C. 20g of germinated noni seeds were ground into a homogenate in a mortar, and the pH was adjusted to 6. The homogenate was filtered through four layers of cheesecloth, and the filtrate was centrifuged at 1000g for 10min to obtain crude cell extract (supernatant). It was then centrifuged at 3500g for 20min. Fractional distillation was performed to obtain crude oil supernatant. The crude oil supernatant was taken and mixed with an equal volume of diethyl ether four times to extract neutral oil. The ether phase was mixed and washed with Na2SO4 aqueous solution to remove peptides. The mixture was concentrated until oil body residue was obtained. The remaining diethyl ether was evaporated under nitrogen, and the composition of the obtained oil was analyzed by thin-layer chromatography. Then, it was used as a substrate to qualitatively test the enzyme activity of lipase.
[0037] In step S3, the pH of the oily residue system is first adjusted to 9. The oily residue is placed in a reaction vessel, and the temperature is controlled at 37°C. 33 mmol / L emulsified gum arabic triolein, 12.8 mmol / L deoxycholate, and 10.7 mmol / L NaCl are added. Then, the lipase activity is measured using p-nitrophenyl palmitate. Based on the measurement results, the immobilized lipase with the highest activity is selected and added to cold-pressed fresh noni seed oil for enzymatic hydrolysis for 5 days. The emulsified gum arabic triolein, deoxycholate, and NaCl are in equal volumes.
[0038] In step S4, after the enzymatic hydrolysis of noni seed oil is completed, the enzymatic hydrolyzed noni seed oil containing the highest activity of lipase is separated and preserved by centrifugation, so that it can be used for the next enzymatic hydrolysis, thus achieving recycling.
[0039] The second aspect of this example provides an enzymatically hydrolyzed noni seed oil with moisturizing and soothing effects.
[0040] Example 2
[0041] This example provides an application of the enzymatically hydrolyzed noni seed oil prepared in Example 1, which is used in a basic face cream.
[0042] The face cream comprises phases A, B, C, and D. Phase A, by mass parts, comprises 2 parts enzymatically hydrolyzed noni seed oil, 1.5 parts glyceryl stearate, and 0.5 parts cetearyl alcohol; Phase B comprises 0.2 parts carbomer 940 and 84.7 parts water; Phase C comprises 0.1 parts a 10% sodium hydroxide aqueous solution; and Phase D comprises 0.5 parts p-hydroxyacetophenone and 0.5 parts 1,2-hexanediol.
[0043] Unless otherwise specified, all of the above-mentioned raw materials are ordinary commercially available products.
[0044] The preparation method of the basic face cream includes:
[0045] S1. Heat phase A to 85℃ and stir until well mixed;
[0046] S2. Heat phase B to 85°C and stir until well mixed;
[0047] S3. Cool phase B to about 50°C, add phase C, and stir evenly (neutralize and thicken, pH around 5.0). Add the pre-dissolved phase D to the thickened phase B, then add it to phase A. Homogenize at 4000 rpm for 3 minutes to obtain the final product.
[0048] Example 3
[0049] This example provides a basic face cream; the face cream includes phase A, phase B, phase C and phase D; phase A, by mass parts, includes 2 parts of unhydrolyzed noni seed oil, 1.5 parts of glyceryl stearate, and 0.5 parts of cetearyl alcohol; phase B includes 0.2 parts of carbomer 940 and 84.7 parts of water; phase C includes 0.1 parts of a 10% sodium hydroxide aqueous solution; phase D includes 0.5 parts of p-hydroxyacetophenone and 0.5 parts of 1,2-hexanediol.
[0050] The preparation method of the basic face cream includes:
[0051] S1. Heat phase A to 85℃ and stir until well mixed;
[0052] S2. Heat phase B to 85°C and stir until well mixed;
[0053] S3. Cool phase B to about 50°C, add phase C, and stir evenly (neutralize and thicken, pH around 5.0). Add the pre-dissolved phase D to the thickened phase B, then add it to phase A. Homogenize at 4000 rpm for 3 minutes to obtain the final product.
[0054] The unhydrolyzed noni seed oil was purchased from Jiangxi Hengcheng Natural Fragrance Oil Co., Ltd.
[0055] Performance testing
[0056] 1. Moisturizing efficacy test
[0057] 1.1. Set the test environment temperature to 20℃-22℃ and relative humidity to 40%-60%.
[0058] 1.2. Testing period: two weeks; Testing area: inner forearm, 2cm×2cm in size.
[0059] 1.3. Test principle: The moisture content of the stratum corneum of the skin is measured based on the capacitance method to verify the moisturizing effect of the sample.
[0060] 1.4. Testing instrument: Multiprobe Moisture Meter D tissue water content measuring instrument, Delfin.
[0061] 1.5. Test results: See Table 1 and Table 2 for details.
[0062] Table 1
[0063]
[0064] Table 2
[0065]
[0066] 1.6. Conclusion: Compared with the control group, the epidermal moisture content of Example 2 (containing 2% enzymatically hydrolyzed noni seed oil) increased significantly, with a 31.14% increase after two weeks; while the moisture content of Example 3 (containing 2% unenzymatically hydrolyzed noni seed oil) only increased by 18.56%.
[0067] In this test, the blank group refers to the blank week, which is the skin moisture content value measured before the sample was used.
[0068] 2. Repair efficacy test
[0069] 2.1. Set the test environment temperature to 20℃-22℃ and relative humidity to 40%-60%.
[0070] 2.2. Testing period: two weeks; Testing area: inner forearm, 2cm x 2cm in size;
[0071] 2.3. Test principle: The erythema value of the sample was measured after physical damage caused by tape tearing and after transdermal water loss, in order to verify the repair efficacy of the sample.
[0072] 2.4. Testing instruments: VapoMeter transdermal moisture loss meter, Delfin; Skin Color Catch skin color meter, Delfin.
[0073] 2.5. Test results: See Tables 3 and 4 for details.
[0074] Table 3
[0075]
[0076] Table 4
[0077]
[0078] 2.6. Conclusion: Compared with the control group, the transdermal water loss in the modeling group was significantly increased, indicating that the model was effective. Compared with the modeling group, both Example 2 (containing 2% enzymatically hydrolyzed noni seed oil) and Example 3 (containing 2% unenzymatically hydrolyzed noni seed oil) significantly reduced transdermal water loss, but Example 1 (containing 2% enzymatically hydrolyzed noni seed oil) was more effective.
[0079] In this test, the blank group and the model group performed a tape tearing experiment. The blank group was the control group before the tape was torn, and the model group was the control group after the tape was torn and before the sample was applied.
[0080] 3. Soothing efficacy test
[0081] 3.1. Set the test environment temperature to 20℃-22℃ and relative humidity to 40%-60%.
[0082] 3.2. Test period: 2 hours; Test area: inner forearm, 2cm x 2cm in size;
[0083] 3.3. Test principle: The erythema value is measured on the physical damage caused by the tape tearing, and then the sample is applied to verify the short-term soothing effect of the sample.
[0084] 3.4. Testing instruments: Delfin; SkinColorCatch skin color measuring instrument, Delfin.
[0085] 3.5. Test results: See Table 5 for details.
[0086] Table 5
[0087]
[0088] 3.6. Conclusion: Compared with the modeling group, both Example 2 (containing 2% enzymatically hydrolyzed noni seed oil) and Example 3 (containing 2% unenzymatically hydrolyzed noni seed oil) significantly alleviated erythema. Example 1 (containing 2% enzymatically hydrolyzed noni seed oil) was better at alleviating erythema than Example 2 (containing 2% unenzymatically hydrolyzed noni seed oil).
[0089] In this test, the modeling group conducted a tape tearing experiment, which specifically refers to the control group before the sample was applied after the tape was torn.
[0090] 4. Emulsification test
[0091] 4.1. Test principle: After enzymatic hydrolysis, long-chain fatty acids are broken down into short-chain fatty acids, which increases emulsifying properties. The difference in the degree of emulsification before and after enzymatic hydrolysis can be compared through emulsification tests and microscopic particle size observation to verify the increased emulsifying ability of noni seed oil after enzymatic hydrolysis.
[0092] 4.2. Test procedure: Mix the noni seed oil before and after enzymatic hydrolysis with water in a certain ratio, let it stand for 1 hour, observe the degree of emulsification with the naked eye, and observe the size of the oil droplets with a microscope.
[0093] 4.3. Testing instruments: CX43 optical microscope, OLYMPUS.
[0094] 4.4. Emulsification performance results: See details. Figure 1 and Figure 2 .
[0095] from Figure 1 The results showed that Example 2 (containing 2% enzymatically hydrolyzed noni seed oil) had better emulsifying properties.
[0096] from Figure 2 The results showed that enzymatically hydrolyzed noni seed oil had a wider emulsification range and smaller, denser particle size.
Claims
1. A method for preparing enzymatically hydrolyzed noni seed oil with moisturizing and soothing effects, characterized in that, Includes the following steps: S1. Prepare fresh noni seeds, wash them, and then germinate them in water. S2. Take out the germinated noni seeds from hydroponics, grind, centrifuge, and fractionate them to separate the oil bodies from the noni seeds that have been hydroponically germinated for 1-7 days, obtain the immobilized lipase in the oil bodies, and test the activity of the immobilized lipase. S3. Select the immobilized lipase with the highest activity for enzymatic hydrolysis of cold-pressed noni seed oil; S4. After enzymatic hydrolysis, separate and preserve the hydrolyzed noni seed oil containing lipase. In step S2, grinding and centrifugation are both carried out at 3-7°C. 10-25g of germinated noni seeds are ground into a homogenate in a mortar, and the pH is adjusted to 5-7. The homogenate is filtered, and the filtrate is centrifuged to obtain a crude cell extract supernatant. The supernatant is then centrifuged and fractionated to obtain a crude oil supernatant. The crude oil supernatant is then mixed with an equal volume of ether four times to extract neutral oil. The ether phase is mixed and washed with Na2SO4 aqueous solution to remove peptides, and concentrated until an oil body is obtained. The remaining ether is evaporated under nitrogen, and the composition of the obtained oil is analyzed by thin-layer chromatography. Then, it is used as a substrate to qualitatively test the enzyme activity of lipase. In step S3, the pH of the oil body system is first adjusted to 7-10. The oil body is placed in a reaction vessel, and the temperature is controlled at 35-43°C. Emulsified gum arabic triolein, deoxycholate, and NaCl are added. The purity of the emulsified gum arabic triolein used in the substrate emulsion is determined by thin-layer chromatography. Then, the lipase activity is determined by p-nitrophenyl palmitate. Based on the measurement results, the immobilized lipase with the highest activity is selected and added to cold-pressed fresh noni seed oil for enzymatic hydrolysis for 1-7 days.
2. An enzymatically hydrolyzed noni seed oil prepared by a method for preparing enzymatically hydrolyzed noni seed oil with moisturizing and soothing effects according to claim 1.
3. An application of enzymatically hydrolyzed noni seed oil with moisturizing and soothing effects according to claim 2, characterized in that, It is used in cosmetics, including face creams.
4. The application of the enzymatically hydrolyzed noni seed oil with moisturizing and soothing effects according to claim 3, characterized in that, The enzymatically hydrolyzed noni seed oil is added to cosmetics at a mass percentage of 0.5-10%.
5. The application of the enzymatically hydrolyzed noni seed oil with moisturizing and soothing effects according to claim 3, characterized in that, The face cream comprises phase A, phase B, phase C, and phase D; phase A comprises enzymatically hydrolyzed noni seed oil, glyceryl stearate, and cetearyl alcohol; phase B comprises carbomer and water; phase C comprises an aqueous solution of sodium hydroxide; and phase D comprises p-hydroxyacetophenone and 1,2-hexanediol.
6. The application of the enzymatically hydrolyzed noni seed oil with moisturizing and soothing effects according to claim 5, characterized in that, The face cream, by weight, comprises: Phase A, 1-3 parts enzymatically hydrolyzed noni seed oil, 1-5 parts glyceryl stearate, and 0.1-2 parts cetearyl alcohol; Phase B, 0.1-1 parts carbomer 940 and 70-90 parts water; Phase C, 0-1 parts a 10% sodium hydroxide aqueous solution; and Phase D, 0-1 parts p-hydroxyacetophenone and 0.1 parts 1,2-hexanediol.
7. The application of the enzymatically hydrolyzed noni seed oil with moisturizing and soothing effects according to claim 6, characterized in that, The method for preparing the face cream includes: S1. Heat phase A and stir until well mixed; S2. Heat phase B and stir until well mixed; S3. Cool down phase B, add phase C, stir evenly, add the pre-dissolved phase D to the thickened phase B, then add it to phase A, homogenize, and you have the final product.