Moisturizing and soothing fermented morinda citrifolia seed oil as well as preparation method and application thereof

Abstract

The invention relates to the field of cosmetics, in particular to the field of A61K8, and particularly relates to moisturizing and soothing fermented morinda citrifolia seed oil as well as a preparation method and application thereof. The preparation method of the fermented morinda citrifolia seed oil comprises the following steps: S1, preparing fresh morinda citrifolia seeds, washing the morinda citrifolia seeds, and performing hydroponic germination on the morinda citrifolia seeds; s2, respectively taking out the germinated morinda citrifolia seeds after 1-7 days, grinding, centrifuging, fractionating, separating oleosomes from the morinda citrifolia seeds after 1-7 days, obtaining immobilized lipase in the oleosomes, and detecting the activity of the immobilized lipase; s3, selecting immobilized lipase with the highest activity for fermenting the cold-pressed morinda citrifolia seed oil; and S4, after fermentation is completed, separating and storing the germinated morinda citrifolia seed oil containing lipase for next reaction oil preparation, and recycling. The fermented morinda citrifolia seed oil obtained through the technology has the moisturizing and soothing effects, and the emulsifying capacity is higher when the fermented morinda citrifolia seed oil is applied to a cosmetic formula.

Description

Technical Field

[0001] The invention relates to the field of cosmetics, in particular to the field of A61K8, and specifically to a moisturizing and soothing fermented noni seed oil, and a preparation method and application thereof. Background Art

[0002] Noni (Morinda citrifolia) is the fruit of the Morinda citrifolia plant. Used as food and medicine for thousands of years in Polynesia, the medicinal uses of the fruit, juice, flowers, leaves, bark, and roots are well known in Polynesia, however, less is known about the potential health benefits of the oil extracted from the seed.

[0003] Noni seeds have long been considered a byproduct of the noni juice industry until the advent of a process that allows the extraction of noni seed oil. The noni fruit contains 200-250 seeds, but it takes tens of thousands of noni seeds to produce 0.024 liters of noni seed oil, making noni seed oil a valuable resource. Although small-scale production of noni seed oil is cost-prohibitive, it is more economically viable when combined with large-scale noni juice production.

[0004] The existing technologies CN103550095A, CN106377453A and CN106214579A disclose the application of noni fruit in cosmetics by using noni fruit extracts, and the skin care effects are all in the direction of whitening, the effects are relatively single, and the production cost is high, which is not conducive to market promotion and practical application. Summary of the invention

[0005] In order to solve the above technical problems, the first aspect of the present invention provides a method for preparing fermented Morinda citrifolia seed oil with moisturizing and soothing effects, comprising the following steps:

[0006] S1. Prepare fresh noni seeds, wash them and then perform hydroponic germination on the noni seeds;

[0007] S2, taking out the 1-7 day old germinated noni seeds respectively, grinding, centrifuging and fractionating to separate the oleosomes in the 1-7 day old noni seeds, obtaining the immobilized lipase in the oleosomes, and detecting the activity of the immobilized lipase therein;

[0008] S3, selecting the most active immobilized lipase for fermenting cold-pressed noni seed oil;

[0009] S4. After the fermentation is completed, the fermented noni seed oil containing lipase is separated and preserved, and the oil can be recycled for the next reaction preparation.

[0010] Furthermore, in step S1, the noni fruit seeds come from noni fruits produced in Hainan.

[0011] Furthermore, the 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 then the pH is adjusted to 5-7; the homogenate is filtered through four layers of cheese cloth, and the filtrate is centrifuged at a centrifugal force of 1000g for 10 minutes to obtain a crude cell extract (supernatant); then centrifuged at a centrifugal force of 3500g for 20 minutes; then fractionated; the crude oil supernatant is taken and mixed with an equal volume of ether four times to extract neutral oils; then mixed with ether, washed with a Na2SO4 aqueous solution to remove polypeptides, and concentrated until oil bodies are obtained; the remaining ether is evaporated under nitrogen, the composition of the obtained oil is analyzed by thin layer chromatography, and then used as a substrate to qualitatively estimate the enzyme activity of the immobilized lipase.

[0012] Furthermore, in the step S3, the pH of the oleosome system is first adjusted to 7-10, the oleosome is placed in a reaction container, the temperature is controlled to be 35-43°C, 33mmol / L of emulsified gum arabic trioleate, 12.8mmol / L of deoxycholate and 10.7mmol / L of NaCl are added; then the lipase activity is determined using p-nitrophenyl palmitate; the immobilized lipase with the highest activity is selected based on the determination results, and is added to cold-pressed fresh noni seed oil for fermentation for 1-7 days; wherein the emulsified gum arabic trioleate, deoxycholate and NaCl are of equal volumes.

[0013] Furthermore, after the fermentation is completed in step S4, the fermented noni seed oil containing immobilized lipase with the highest activity is separated by centrifugation and preserved, which can be used for the next fermentation to achieve recycling.

[0014] More preferably, the hydroponic germination time of the noni fruit seeds in step S1 is 4-6 days.

[0015] Further preferably, the S2 grinding and centrifugation are both carried out at 4-6°C, 15-20g of germinated noni seeds are ground into a homogenate in a mortar, and then the pH is adjusted to 6; the homogenate is filtered through four layers of cheese cloth, and the filtrate is centrifuged at a centrifugal force of 1000g for 10 minutes to obtain a crude cell extract (supernatant); the extract is then centrifuged at a centrifugal force of 3500g for 20 minutes; fractionation is then performed; the crude oil supernatant is taken and mixed with an equal volume of ether four times to extract neutral oils; the ether is mixed, washed with a Na2SO4 aqueous solution to remove polypeptides, and concentrated until oleosomes are obtained; the remaining ether is evaporated under nitrogen, and the composition of the obtained oil is analyzed by thin layer chromatography, and then used as a substrate to qualitatively estimate the enzyme activity of the immobilized lipase.

[0016] More preferably, in the step S3, the pH of the oleosome system is first adjusted to 8-9, the oleosome is placed in a reaction vessel, the temperature is controlled to 36-40°C, 33 mmol / L of emulsified gum arabic trioleate, 12.8 mmol / L of deoxycholate and 10.7 mmol / L of NaCl are added; the purity of the emulsified gum arabic trioleate used in the substrate emulsion is determined by thin layer chromatography; then the lipase activity is determined by p-nitrophenyl palmitate; the immobilized lipase with the highest activity is selected according to the determination results, and added to the cold-pressed fresh noni seed oil for fermentation for 3-5 days.

[0017] The second invention of the present invention provides a fermented noni seed oil prepared by the above preparation method.

[0018] The third invention of the present invention provides the application of fermented noni seed oil in cosmetics.

[0019] Preferably, the cosmetics include one or more of essence, emulsion, cream, ointment and mask.

[0020] Preferably, the fermented noni seed oil is added to the cosmetic in an amount of 0.5-10% by mass.

[0021] Beneficial Effects

[0022] (I) The present invention separates oleosomes from germinated noni seeds, obtains the oleosome components of lipase embedded in the natural noni seed oleosomes, and uses them as immobilized lipase in fermenting cold-pressed noni seed oil.

[0023] (ii) The present invention detects the activity of immobilized lipase in oleosomes isolated from noni seeds germinated for 1-7 days, and selects the best oleosomes from germinated seeds for fermentation and cold-pressing noni seed oil.

[0024] (III) After the fermentation is finished, the present invention separates the highly active immobilized lipase oleosomes and the fermented noni seed oil after the reaction, and continues to serve as the source of enzymolysis products for the next reaction and is recycled.

[0025] (iv) The fermented noni seed oil obtained by the process of the present invention has moisturizing and soothing effects, and has stronger emulsification ability when used in formulas.

[0026] (V) The present invention utilizes fixed lipase oleosomes extracted from germinated noni seeds to ferment cold-pressed fresh noni seed oil. Compared with unfermented noni seed oil, the fermented noni seed oil has higher active ingredients and better efficacy. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is the emulsification performance test result diagram.

[0028] Figure 2 This is the oil droplet morphology test result diagram. DETAILED DESCRIPTION

[0029] Example 1

[0030] The first aspect of this example provides a method for preparing fermented noni seed oil with moisturizing and soothing effects, which comprises the following steps:

[0031] S1. Prepare fresh noni seeds, wash them and then perform hydroponic germination on the noni seeds;

[0032] S2, taking out the 1-7 day old germinated noni seeds respectively, grinding, centrifuging and fractionating to separate the oleosomes in the 1-7 day old noni seeds, obtaining the immobilized lipase in the oleosomes, and detecting the activity of the immobilized lipase therein;

[0033] S3, selecting the most active immobilized lipase for fermenting cold-pressed noni seed oil;

[0034] S4. After the fermentation is completed, the fermented noni seed oil containing lipase is separated and preserved, and the oil can be recycled for the next reaction preparation.

[0035] In step S1, the noni fruit seeds come from noni fruit produced in Hainan, and the hydroponic germination time is 5 days.

[0036] The S2 grinding and centrifugation are both carried out at 5°C. 20g of germinated noni seeds are ground into a homogenate in a mortar, and then the pH is adjusted to 6; the homogenate is filtered through four layers of cheese cloth, and the obtained filtrate is centrifuged at a centrifugal force of 1000g for 10 minutes to obtain a crude cell extract (supernatant); the filtrate is then centrifuged at a centrifugal force of 3500g for 20 minutes; fractionation is performed to obtain a crude supernatant; the crude supernatant is taken and mixed with an equal volume of ether four times to extract neutral oils; the ether is mixed, washed with a Na2SO4 aqueous solution to remove polypeptides, and concentrated until an oleosome residue is obtained; the remaining ether is evaporated under nitrogen, and the composition of the obtained oil is analyzed by thin layer chromatography, and then used as a substrate for qualitative estimation of the enzyme activity of lipase.

[0037] In the step S3, the pH of the oleosome residue system is first adjusted to 9, the oleosome residue is placed in a reaction container, the temperature is controlled to 37°C, 33 mmol / L of emulsified gum arabic trioleate, 12.8 mmol / L of deoxycholate and 10.7 mmol / L of NaCl are added; then the lipase activity is determined using p-nitrophenyl palmitate; the immobilized lipase with the highest activity is selected based on the determination results, added to cold-pressed fresh noni seed oil, and fermented for 5 days; wherein the emulsified gum arabic trioleate, deoxycholate and NaCl are equal in volume.

[0038] In the step S4, after the fermentation is completed to obtain the fermented noni seed oil, the fermented noni seed oil containing lipase with the highest activity is separated by centrifugation and preserved, which can be used for the next fermentation to achieve recycling.

[0039] The second aspect of this example provides a fermented noni seed oil with moisturizing and soothing effects.

[0040] Example 2

[0041] This example provides an application of the fermented noni seed oil prepared in Example 1, which is applied in a basic facial cream.

[0042] The facial cream comprises phase A, phase B, phase C and phase D; phase A comprises 2 parts of fermented noni seed oil, 1.5 parts of glyceryl stearate, and 0.5 parts of cetearyl alcohol in terms of weight; phase B comprises 0.2 parts of carbomer 940 and 84.7 parts of water; phase C comprises 0.1 parts of a 10% sodium hydroxide aqueous solution; and phase D comprises 0.5 parts of p-hydroxyacetophenone and 0.5 parts of 1,2-hexanediol.

[0043] Unless otherwise specified, the above raw materials are common commercially available products.

[0044] The preparation method of the basic facial cream comprises:

[0045] S1. Heat phase A to 85°C and stir to mix evenly;

[0046] S2. Heat phase B to 85°C and stir to mix evenly;

[0047] S3. Cool phase B to about 50°C, add phase C, and stir evenly (neutralization and thickening, pH is about 5.0). Add the pre-dissolved phase D to the thickened phase B, and then add it to phase A. Homogenize at 4000rpm for 3min.

[0048] Example 3

[0049] This example provides a basic facial cream; the facial cream includes phase A, phase B, phase C and phase D; phase A includes 2 parts of unfermented noni seed oil, 1.5 parts of glyceryl stearate, and 0.5 parts of cetearyl alcohol in terms of weight; 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 facial cream comprises:

[0051] S1. Heat phase A to 85°C and stir to mix evenly;

[0052] S2. Heat phase B to 85°C and stir to mix evenly;

[0053] S3. Cool phase B to about 50°C, add phase C, and stir evenly (neutralization and thickening, pH is about 5.0). Add the pre-dissolved phase D to the thickened phase B, and then add it to phase A. Homogenize at 4000rpm for 3min.

[0054] The unfermented Noni seed oil was purchased from Jiangxi Hengcheng Natural Spice Oil Co., Ltd.

[0055] Performance Testing

[0056] 1. Moisturizing effect test

[0057] 1.1. Set the test environment temperature: 20℃-22℃, relative humidity: 40%-60%.

[0058] 1.2. Test period: two weeks; Test area: inner side of 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. Test instrument: Multiprobe Moisture Meter D tissue moisture content measuring instrument, Delfin.

[0061] 1.5. Test results: See Table 1 and Table 2 for details.

[0062] Table 1

[0063] Skin moisture content Blank Week Week 1 Week 2 Example 2 28.9 38.7 37.9 Example 3 29.1 35.2 34.5

[0064] Table 2

[0065] Moisture content improvement rate (%) Week 1 Week 2 Example 2 33.9％ 31.14％ Example 3 20.96％ 18.56％

[0066] 1.6. Conclusion: Compared with the blank group, the epidermal moisture content of Example 2 (containing 2% fermented Noni seed oil group) increased significantly, and the moisture content increased by 31.14% after two weeks; while that of Example 3 (containing 2% unfermented Noni seed oil group) only increased by 18.56%.

[0067] The blank group in this test refers to the blank week, that is, the skin moisture content value measured before using the sample.

[0068] 2. Repair efficacy test

[0069] 2.1. Set the test environment temperature: 20℃-22℃, relative humidity: 40%-60%;

[0070] 2.2. Test period: two weeks; Test area: inner side of forearm, 2cm×2cm;

[0071] 2.3. Test principle: Measure the erythema value of the physical damage caused by tearing the tape and measure it again after applying the sample after transepidermal water loss to verify the repair effect of the sample.

[0072] 2.4. Test instruments: VapoMeter transepidermal water loss meter, Delfin; Skin Color Catch skin color meter, Delfin.

[0073] 2.5. Test results: See Table 3 and Table 4 for details.

[0074] Table 3

[0075] Transepidermal water loss Modeling Group Week 1 Week 2 Example 2 54 18.43 10.37 Example 3 59 26.30 17.36

[0076] Table 4

[0077] Transepidermal water loss improvement rate (%) Week 1 Week 2 Example 2 65.87 80.80 Example 3 55.42 70.57

[0078] 2.6. Conclusion: Compared with the blank group, the degree of transepidermal water loss in the modeling group increased significantly, indicating that the model establishment was effective; compared with the modeling group, Example 2 (containing 2% fermented noni seed oil) and Example 3 (containing 2% unfermented noni seed oil) could significantly reduce transepidermal water loss, but Example 1 (containing 2% fermented noni seed oil) had a better effect.

[0079] In this test, the blank group and the modeling group performed the tape tearing experiment. The blank group was the control group before the tape was torn off, and the model group was the control group after the tape was torn off but before the sample was painted.

[0080] 3. Soothing effect test

[0081] 3.1. Set the test environment temperature: 20℃-22℃, relative humidity: 40%-60%;

[0082] 3.2. Test period: 2h; Test area: inner side of forearm, 2cm×2cm;

[0083] 3.3. Test principle: The erythema value is measured for the physical damage caused by tearing the tape, and then the sample is applied to verify the soothing effect of the sample in the short term.

[0084] 3.4. Test instrument: Delfin; SkinColorCatch skin color measuring instrument, Delfin.

[0085] 3.5. Test results: See Table 5 for details.

[0086] Table 5

[0087] Δ erythema value 10min 30min 90min 150min Example 2 1.00 6.67 9.00 9.67 Example 3 0.33 3.33 6.33 7.00

[0088] 3.6. Conclusion: Compared with the modeling group, Example 2 (containing 2% fermented noni seed oil) and Example 3 (containing 2% unfermented noni seed oil) can significantly alleviate erythema, and Example 1 (containing 2% fermented noni seed oil) has better erythema relief ability than Example 2 (containing 2% unfermented noni seed oil).

[0089] In this test, the modeling group performed a tape tearing experiment, which specifically refers to the control group after the tape was torn off but before the sample was painted.

[0090] 4. Emulsification test

[0091] 4.1. Test principle: After fermentation, long-chain fatty acids are broken into short-chain fatty acids, and the emulsification performance is increased. The difference in emulsification degree before and after fermentation can be compared through emulsification test and particle size observation to verify the increase in the emulsification capacity of noni seed oil after fermentation.

[0092] 4.2. Test steps: Mix the noni seed oil before and after fermentation with water in a certain proportion, let it stand for 1 hour, observe the emulsification degree with the naked eye and observe the oil droplet size with a microscope.

[0093] 4.3. Test instrument: CX43 optical microscope, OLYMPUS.

[0094] 4.4. Emulsification performance results: see Figure 1 and Figure 2 .

[0095] from Figure 1 The results show that Example 2 (containing 2% fermented noni seed oil) has better emulsification performance. Figure 2 The results show that fermented noni seed oil has a wider emulsification range and smaller and denser particle size.

Claims

1. A method for preparing fermented noni seed oil with moisturizing and soothing effects, characterized in that: The following steps are involved: S1. Prepare fresh noni seeds, wash them and then perform hydroponic germination on the noni seeds; S2, taking out the hydroponic germinated noni seeds, grinding, centrifuging, and fractionating to separate the oleosomes in the noni seeds of 1-7 days old, obtaining the immobilized lipase in the oleosomes, and detecting the activity of the immobilized lipase therein; S3, selecting the most active immobilized lipase for fermenting cold-pressed noni seed oil; S4. After the fermentation is completed, the fermented noni seed oil containing lipase is separated and preserved.

2. The method for preparing the fermented noni seed oil with moisturizing and soothing effects according to claim 1, characterized in that: The hydroponic germination time of the noni seeds in step S1 is 1-7 days.

3. The method for preparing the fermented noni seed oil with moisturizing and soothing effects according to claim 2, characterized in that: The 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 then the pH is adjusted to 5-7; the homogenate is filtered, and the obtained filtrate is centrifuged to obtain a crude cell extract supernatant; the crude supernatant is then centrifuged and fractionated to obtain a crude oil supernatant; the crude oil supernatant is taken and mixed with an equal volume of ether four times to extract neutral oil; the ether is mixed, washed with a Na2SO4 aqueous solution to remove polypeptides, and concentrated until an oleosome is obtained; the remaining ether is evaporated under nitrogen, and the composition of the obtained oil is analyzed by thin layer chromatography, and then used as a substrate to qualitatively estimate the enzyme activity of lipase.

4. The method for preparing the fermented noni seed oil with moisturizing and soothing effects according to claim 3, characterized in that: In the step S3, the pH of the oleosome system is first adjusted to 7-10, the oleosome is placed in a reaction container, the temperature is controlled to be 35-43° C., emulsified gum arabic trioleate, deoxycholate and NaCl are added; the purity of the emulsified gum arabic trioleate used in the substrate emulsion is determined by thin layer chromatography; then the lipase activity is determined by p-nitrophenyl palmitate; the immobilized lipase with the highest activity is selected according to the determination result, and is added to the cold-pressed fresh noni seed oil for fermentation for 1-7 days.

5. A fermented noni seed oil prepared according to the method for preparing the fermented noni seed oil with moisturizing and soothing effects according to any one of claims 1 to 4.

6. An application of the fermented noni seed oil with moisturizing and soothing effects according to claim 5, characterized in that: Used in cosmetics, including facial creams.

7. The use of fermented noni seed oil with moisturizing and soothing effects according to claim 6, characterized in that: The fermented noni seed oil is added to cosmetics in an amount of 0.5-10% by mass.

8. The use of fermented noni seed oil with moisturizing and soothing effects according to claim 6, characterized in that: The facial cream comprises phase A, phase B, phase C and phase D; phase A comprises fermented noni seed oil, glyceryl stearate and cetearyl alcohol; phase B comprises carbomer and water; phase C comprises sodium hydroxide aqueous solution; and phase D comprises p-hydroxyacetophenone and 1,2-hexanediol.

9. The use of fermented noni seed oil with moisturizing and soothing effects according to claim 8, characterized in that: The facial cream comprises, by weight, phase A comprising 1-3 parts of fermented noni seed oil, 1-5 parts of glyceryl stearate, and 0.1-2 parts of cetearyl alcohol; phase B comprising 0.1-1 parts of carbomer 940 and 70-90 parts of water; phase C comprising 0-1 parts of a 10% sodium hydroxide aqueous solution; and phase D comprising 0-1 parts of p-hydroxyacetophenone and 0.1 parts of 1,2-hexanediol.

10. The use of fermented noni seed oil with moisturizing and soothing effects according to claim 8, characterized in that: The preparation method of the facial cream comprises: S1. Heat phase A and stir to mix evenly; S2, heat phase B and stir to mix evenly; S3. Cool down phase B, add phase C, stir evenly, add the pre-dissolved phase D to the thickened phase B, and then add it to phase A, homogenize, and you have it.

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