A freeze-dried essence and preparation method thereof

Abstract

The present invention discloses a freeze-dried essence and a preparation method thereof, belonging to the field of cosmetics. The freeze-dried essence of the present invention comprises a solution component A and a freeze-dried powder component B; the solution component A comprises a lily seed extract and a panax notoginseng extract; the freeze-dried powder component B comprises fibronectin; the mass ratio of the lily seed extract, the panax notoginseng extract and the fibronectin is (0.01-0.03): (0.005-0.015): 1. By combining the synergistic components of fibronectin and the plant composition, the symptoms of skin oxidation relaxation and wrinkling, skin dehydration, melanin deposition, acne inflammation and the like caused by "hormone face" and related inducements can be effectively improved at the same time.

Description

Technical Field

[0001] The invention relates to the field of cosmetics, and in particular to a freeze-dried essence and a preparation method thereof. Background Art

[0002] As people are under increasing work pressure and pay less and less attention to skin care, more and more skin problems are appearing, especially for people with sensitive skin. After being stimulated by certain external factors, the skin is prone to dehydration, acne, inflammation, allergies, stratum corneum damage, enlarged pores and other complications. Although there are many cosmetics on the market that can repair and maintain the skin, their functions are relatively single.

[0003] For sensitive skin, the most common symptom is "hormone face", which is mainly manifested by the appearance of spots, red blood streaks and other problems when the skin is slightly stimulated. In response to this situation, people have found that the use of fibronectin as an active ingredient has a better therapeutic effect. This substance can shrink and close the pores of the skin through the supplement of exogenous protein, inhibiting the impact of external stimuli on the skin. At the same time, it has a certain repair effect on the stratum corneum cells of the skin, and has a good improvement on the common "hormone face" symptoms.

[0004] However, there are many causes of "hormone face". In addition to the symptoms of "hormone face", these causes generally also cause other different types of skin problems, such as a surge in skin wrinkles, skin dehydration, melanin deposition, and acne inflammation. Fibronectin alone cannot relieve these symptoms at the same time. Summary of the invention

[0005] Based on the defects of the prior art, the purpose of the present invention is to provide a freeze-dried essence, which, through the synergistic components of fibronectin and a plant composition, can effectively improve the symptoms of "hormone face" and related causes such as skin oxidative relaxation and wrinkling, skin dehydration, melanin deposition, acne inflammation, etc.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is:

[0007] A freeze-dried essence, comprising a solution component A and a freeze-dried powder component B;

[0008] The solution component A includes Litsea cubeba extract and Panax notoginseng extract; the freeze-dried powder component B includes fibronectin;

[0009] The mass ratio of the Litsea cubeba seed extract, Panax notoginseng extract and fibronectin is (0.01-0.03):(0.005-0.015):1.

[0010] Since fibronectin cannot be dissolved in conventional solvents and has high biological activity, it is easily oxidized and deteriorated during processing and use. Therefore, the inventor of this scheme introduces fibronectin into the product in the form of freeze-dried powder. The main effect of fibronectin on symptoms such as "hormone face" is to replenish exogenous proteins and attach to the surface of skin cells to regulate cell polarity, powderization and growth, effectively enhance the enthusiasm of cell differentiation, movement and self-healing, and effectively shrink skin pores to protect them from secondary stimulation. However, prior art research has found that the improvement effect of fibronectin is very limited, mainly because fibronectin mostly acts on the surface of the skin and is consumed quickly, and cannot achieve long-term and deep skin problem improvement. At the same time, based on its mode of action, it cannot work well for some inflammatory symptoms caused by dehydration, melanin deposition and secretion disorders in the deep skin. Based on the above reasons, the inventors introduced the highly active Litsea cubeba extract and Panax notoginseng extract in the form of a solution together with fibronectin. However, during the use of this product, it is only necessary to mix the solution component and the freeze-dried powder component to form a highly active fibronectin carrier and use it directly on the skin to achieve improvement of multiple skin symptoms.

[0011] Among them, Litsea cubeba seed, also known as Litsea cubeba, contains more active ingredients (such as citral). In the prior art, it is mainly used as a raw material for preparing flavors. In the field of efficacy, it is mainly used as a traditional Chinese medicine, with the effects of invigorating the spleen and appetite, clearing away heat and activating blood circulation, etc., and is rarely mentioned as an active ingredient in cosmetics; Panax notoginseng is also a traditional Chinese medicine, which is rich in ginsenosides and has great value in disease treatment (especially tumors), and Panax notoginseng extracts are also used in some cosmetics, which can show certain effects such as whitening and anti-aging. When the inventors matched the two plant extract components with fibronectin in a specific ratio, they found that the two plant extracts can effectively assist fibronectin to enter the deep layer of the skin to further improve the symptoms of "hormone face", and at the same time have a good synergistic effect: release active ingredients to enhance the activity of antioxidant enzymes contained in the skin, assist in the removal of free radicals, and cooperate with the cell mobilization of fibronectin, so that the cells can fully absorb external moisture, and finally play a long-term deep wrinkle removal and moisturizing effect; synergistically inhibit the expression of inflammatory factors existing in a variety of skins, and at the same time promote the excretion of inflammatory products by cells, and finally play a short-term and efficient acne removal and anti-inflammatory mechanism; synergistically inhibit the activity of melanin-related tyrosinase in the deep skin, effectively improve the melanin precipitation in the deep skin, and also play a certain physical sunscreen effect, playing a whitening effect on the skin from both internal and external dual effects. At the same time, in the matching process, based on the complex mechanism of multiple effects, the type and ratio selection of the two plant extracts also have a high degree of specific matching. If the type or ratio is replaced, a more ideal effect cannot be achieved.

[0012] Preferably, the mass ratio of the Litsea cubeba extract, Panax notoginseng extract and fibronectin is 0.02:0.01:1.

[0013] The inventors have discovered through experiments that the effective range for achieving synergistic effects when the three are combined is very narrow. Only within a specific range can these three components take into account the improvement of "hormone face" and multiple symptoms such as increased skin wrinkles, skin dehydration, melanin deposition, and acne inflammation, and effectively improve the degree of improvement. If the ratio is not good, it is likely to lead to a low degree of improvement, or the inability to achieve multiple effects. The three components in the above optimal ratio have the best synergistic effect.

[0014] Preferably, the freeze-dried powder component B also contains D-mannitol, crystalline trehalose, soybean polypeptide and acetyl hexapeptide-1.

[0015] In order to ensure the stability of fibronectin in the freeze-dried powder component B, and also to ensure that the freeze-dried powder component B as a whole has good binding compatibility when mixed with the solution component A during use, a variety of functional excipients are introduced into the freeze-dried powder component B, among which D-mannitol, crystalline trehalose, etc. have good waterproof and hygroscopic effects, and soybean polypeptides and acetyl hexapeptide-1 also have good antioxidant properties. At the same time, each component has good compatibility when in contact with the solution components, and can also assist fibronectin in achieving moisturizing and antioxidant functions on the skin, thereby enhancing the degree of improvement of skin defects.

[0016] More preferably, the freeze-dried powder component B contains the following components in parts by weight: 4-6 parts of D-mannitol, 0.4-0.6 parts of crystalline trehalose, 0.4-0.6 parts of soybean polypeptide, 0.05-0.15 parts of acetyl hexapeptide-1 and 0.8-1.5 parts of fibronectin.

[0017] Preferably, the solution component A also contains a moisturizer, an antioxidant and a preservative.

[0018] More preferably, the moisturizer is at least one of hyaluronic acid and Tremella fuciformis extract; the antioxidant is at least one of superoxide dismutase and carnosine; and the preservative is at least one of cinnamon and hexylene glycol.

[0019] When human skin is symptomatic, dehydration will lead to further deterioration of the condition. Therefore, introducing a certain amount of moisturizer into solution component A can effectively inhibit the deterioration of skin symptoms and provide a better working environment for Litsea cubeba extract, Panax notoginseng extract and fibronectin. At the same time, superoxide dismutase (SOD) is a key antioxidant metal enzyme in the body, and carnosine is also a non-toxic antioxidant with significant anti-aging effects. It can inhibit the glycation of collagen and the production of AGEs in the enzyme metabolic pathway. Combining the two with Litsea cubeba extract and Panax notoginseng extract can effectively enhance the activity of skin cells, reduce cell loss and maintain moisture in skin cells.

[0020] More preferably, the solution component A includes the following components in parts by weight: 0.8-1.2 parts of hyaluronic acid or tremella extract, 2.5-3.5 parts of superoxide dismutase, 0.1-0.15 parts of carnosine, 0.5-1 parts of cinnamomeone, 0.5-1 parts of hexylene glycol, 0.01-0.03 parts of Litsea cubeba seed extract and 0.005-0.015 parts of Panax notoginseng extract.

[0021] Another object of the present invention is to provide a method for preparing the freeze-dried essence, comprising the following steps:

[0022] (1) mixing the components of freeze-dried powder component B and adding water to mix evenly, followed by freeze drying and sieving to obtain freeze-dried powder component B;

[0023] (2) Mix the components of solution component A and add water to mix evenly to obtain solution component A.

[0024] The preparation method of the freeze-dried essence of the present invention has simple operation steps, low requirements on equipment, and the prepared product has strong stability and is not easy to deteriorate, and can realize industrial large-scale production.

[0025] The beneficial effect of the present invention is that the present invention provides a freeze-dried essence, which, through the synergistic components of fibronectin and a plant composition, can effectively improve the symptoms of "hormone face" and skin oxidation relaxation and wrinkling, skin dehydration, melanin deposition, acne inflammation and the like caused by related factors. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is a graph showing the results of the anti-inflammatory and soothing efficacy test in Effect Example 1 of the present invention. DETAILED DESCRIPTION

[0027] In order to better illustrate the purpose, technical scheme and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments and comparative examples, the purpose of which is to understand the content of the present invention in detail, rather than to limit the present invention. All other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. The experimental reagents and instruments involved in the implementation of the present invention, unless otherwise specified, are all commonly used ordinary reagents, raw materials and instruments. Among them, fibronectin can be directly used as a commercially available product, or it can be an artificial homemade product, as long as it can achieve a functional effect similar to that of existing fibronectin. The preparation process of the product includes fermentation, centrifugal collection of bacteria, homogenization and crushing of bacteria, centrifugal separation of crushed solution, chromatography, purification and filtration and other steps.

[0028] Examples 1 to 9

[0029] The freeze-dried essence and the preparation method thereof according to the present invention are described in an embodiment. The raw material components of the freeze-dried essence are shown in Table 1.

[0030] The preparation method of the freeze-dried essence comprises the following steps:

[0031] (1) The raw materials of freeze-dried powder component B in Table 1 are mixed uniformly, then freeze-dried and sieved to obtain freeze-dried powder component B;

[0032] (2) The components of solution component A are mixed uniformly to obtain solution component A. The weight units of the components of solution component A are the same as those of freeze-dried powder component B. When the product is used, the two components are mixed in equal amounts.

[0033] Comparative Examples 1 to 6

[0034] The difference between the comparative examples and the embodiments is only in the types and proportions of components, as shown in Table 2.

[0035] Unless otherwise specified, the components and raw materials used in the embodiments and comparative examples of the present invention are all commercially available raw materials, and the components and raw materials used in each parallel experiment are all of the same kind.

[0036] Table 1

[0037]

[0038] Table 2

[0039]

[0040] Effect Example 1

[0041] In order to verify the performance of the freeze-dried essence of the present invention, the products of each embodiment and / or comparative example were subjected to the following performance tests, and the specific steps are as follows:

[0042] (1) Anti-inflammatory and soothing efficacy test: Anti-inflammatory detection based on LPS-induced RAW264.7 cell inflammation model, Model (model group): 200ul PBS and LPS (final concentration 1ug / ml) were added to a 6-well plate; Control (blank control group): 200ul PBS was added to a 6-well plate; Test sample (Example 1 product): 200ul filtered test sample and LPS (final concentration 1ug / ml) were added to a 6-well plate; 3 replicate wells were set for each treatment group, and cell supernatants were collected after incubation for 24 hours. The operation was performed according to the instructions of the ELISA kit (the dosage concentration of the test sample was 0.25 times that of the model group), and the test results of the inflammatory factor concentration test of the test sample and the model were compared. If the inflammatory factor concentration of the test sample was lower than that of the model and the statistical difference P value was <0.05, it can be considered that the test sample has anti-inflammatory and soothing efficacy; the test results are as follows Figure 1 and as shown in Table 1 , where *** indicates P < 0.001 compared with Model;

[0043] (2) Skin cell density test before and after use: 3D full-thickness skin model provided by Guangdong Boxi Biotechnology Co., Ltd. According to the experimental grouping and experimental treatment conditions (see Table 2), the SSUV irradiation treatment (UVA: 30J / cm 2 ; UVB: 50mJ / cm 2 ), then a certain amount of samples (5wt% of the product in Example 1, positive control sample) were applied to the corresponding model surface, and applied every other day. The total duration was 4 days. Finally, the model samples were fixed, embedded and sliced, and H&E staining (tissue morphology analysis) and IHC (COL-Ⅳ) staining analysis were performed. Image-Pro Plus software was used to statistically analyze the IOD value (Integrated optical density) of the yellow-brown area at the junction of the true epidermis in each group of photos after IHC staining. The IOD value can indirectly reflect the expression of COL-Ⅳ; the IOD value of the purple cell nucleus area in each group of photos after H&E staining was statistically analyzed, and the IOD value can indirectly reflect the cell density; the test results are shown in Table 3; when the t-test method is used for statistics, the significance of the negative control compared with the positive control is represented by #, and if p < 0.001, it is represented by ###; the significance of the positive control, the sample to be tested and the negative is represented by *, and if p < 0.01, it is represented by **, and if p < 0.001, it is represented by ***;

[0044] (3) DPPH free radical scavenging test: The positive control was dissolved and diluted with 95% ethanol to a series of concentration gradients of 0.08 mg / mL, 0.04 mg / mL, 0.02 mg / mL, and 0.01 mg / mL to verify the test system. Test substance treatment: The test substance was diluted with water to form a multi-level concentration sample. Referring to Table 4, 10 mL test tubes were used to set up the sample tube (T), sample background (T0), DPPH tube (C), and solvent background (C0). Three parallel tubes were required for each sample tube (T) of each tested concentration, and three parallel tubes were also required for the DPPH tube (C). 1 mL of the same amount of sample solution was added to each sample tube (T) and sample background (T0). Solvent water was added to all test tubes (T, T0, C, C0) to a volume of 3 mL and mixed. Add 1 mL of DPPH ethanol solution to the sample tube (T) and DPPH tube (C), replace the sample background (T0) and solvent background (C0) with 95% ethanol, shake gently, and let stand at room temperature for 5 minutes. Transfer each reaction solution into a 1 cm cuvette, measure the absorbance at 517 nm, and the reagent dosage is shown in Table 4. Measure the absorbance at a wavelength of 517 nm and calculate the DPPH removal rate CR (%) of the sample to be tested, where the calculation formula of CR (%) is:

[0045]

[0046] Where T is the absorbance value of the sample tube, that is, the absorbance value of the solution after the sample reacts with DPPH;

[0047] T0 is the absorbance value of the sample background;

[0048] C is the average absorbance of three tests of the DPPH tube, that is, the average absorbance of the DPPH solution when no sample is added;

[0049] C0 is the absorbance value of the solvent background;

[0050] The test results are shown in Table 5;

[0051] (4) Test for inhibition of protein non-enzymatic glycosylation: 1.7 g of bovine serum albumin and 1.0 g of glyoxal were added to 100 mL of phosphate buffer, and 0.2 g of sodium azide was added, and the mixture was shaken to form a reaction solution. The mixture was incubated at 37°C for 72 h. The optimal fluorescence detection conditions for the product were an excitation wavelength of 440 nm, a slit of 10 nm, an emission wavelength of 480 nm, a slit of 10 nm, and a detection range of 460 to 600 nm. The production of AGEs was detected by fluorescence. The reagent addition conditions are shown in Table 6 (where "+" indicates addition, and the added volume is 2 mL; "-" indicates no addition, the negative control group is phosphate buffer PBS, the positive control group is a mixture of vitamin C and ethyl ether, and the positive control and sample groups are directly taken in the same volume). The ability of the sample to inhibit the AGEs generation process was tested and the inhibition rate IR (%) was calculated. The calculation formula of IR (%) is:

[0052]

[0053] The test results are shown in Table 7, where SD is the standard deviation;

[0054] (5) Acne test before and after skin use: 16 groups of volunteers aged 30±8 years old were selected for the test, with 25 people in each group, half of whom were male and half were female. All volunteers had facial acne caused by non-special circumstances. Except for one group of people who used a placebo (colored gel solution), the other groups used the products of Examples 1 to 9 and Comparative Examples 1 to 6 respectively, and applied them on the acne lesions for 7 days. After 7 days, the effective rate (the affected area was significantly improved) and the cure rate (the affected area was completely cured) were calculated, and the results were statistically analyzed as shown in Table 8.

[0055] Table 1

[0056]

[0057] It can be seen from Table 1 that the freeze-dried essence prepared in Example 1 of the present invention can effectively inhibit the expression activity of inflammatory factors caused by external factors, and thus has good anti-inflammatory and soothing effects.

[0058] Table 2

[0059]

[0060] Table 3

[0061]

[0062] It can be seen from Table 3 that the freeze-dried essence prepared in Example 1 of the present invention can alleviate the problem of decreased skin cell density caused by external factors in human skin, increase skin cell density, and ensure skin firmness.

[0063] Table 4

[0064] Add reagent (mL) T-sample tube <![CDATA[T0 - Sample background]]> C-DPPH tube <![CDATA[C0 - Solvent Background]]> Sample solution (mL) 1 1 - - Water solvent (mL) 2 2 3 3 DPPH ethanol solution (mL) 1 - - - 95% ethanol (mL) - 1 - 1

[0065] Table 5

[0066]

[0067]

[0068] As can be seen from Table 5, the products prepared in each embodiment of the present invention have a good DPPH scavenging rate, which can reach more than 55%. From the comparison of Examples 1 to 9, it can be seen that the addition ratio of the two key extracts in the product components is different, and the DPPH scavenging effect produced is different, especially the extract of Litsea cubeba, the change of its content has a great influence on the performance of the product. If the ratio of the two plant extracts is improper or only one of the plant extracts is selected, it may not be able to achieve effective scavenging performance as shown in Comparative Examples 1 to 3; on the other hand, if these two are replaced with other plant extracts, the scavenging performance may be poor as shown in Comparative Examples 4 or 5. As can be seen from the comparison of Examples 6 to 9, the differences in other components in the freeze-dried essence will also have a certain impact on the performance of the product.

[0069] Table 6

[0070] Add reagents Test samples Positive PBS Reactants Sample set + - - + Negative control group - - + + Positive control group - + - + Blank control group + - + -

[0071] Table 7

[0072]

[0073] As can be seen from Table 7, although the inhibition rate of the freeze-dried essence prepared in Example 1 of the present invention on protein non-enzymatic glycosylation is not as good as that of the positive control group in the early stage, as time goes by, its inhibition rate gradually increases and exceeds that of the positive control group, indicating that the freeze-dried essence of the present invention has a good AGEs generation inhibitory effect, and thus has a beneficial effect on the anti-aging and whitening of the skin.

[0074] Table 8

[0075]

[0076]

[0077] As can be seen from Table 8, the products prepared in the embodiments of the present invention have acne removal effects, especially the product of Example 1, which has an effective rate of more than 90% after use and a cure rate of more than 60%. In contrast, Comparative Example 1 does not contain Panax notoginseng extract, and although the acne removal efficiency is high, it cannot achieve a good cure effect relatively speaking; while Comparative Example 2 does not contain Litsea cubeba extract, and its effective rate and cure rate are both low, indicating that in the freeze-dried essence of the present invention, the two plant extracts need to be in a synergistic relationship to achieve the expected sustained therapeutic effect; the two in the product of Comparative Example 3 are improperly matched, and its acne removal effect is significantly reduced compared to the product of Example 1, and the products of Comparative Examples 5 and 6 replace the two key components with ginseng extract and aloe extract respectively, and such a combination does not play the expected synergistic effect, and the acne removal efficiency and cure rate are low.

[0078] Effect Example 2

[0079] In order to verify the preservation performance and skin irritation of the freeze-dried essence of the present invention, the product of Example 1 was subjected to stability test and skin irritation (safety) test: the test was carried out by using a classic patch test, specifically, the product of Example 1 was used as the test sample, the blank control was used as the negative control, and subjects aged 30±8 years (13 males and 17 females) were selected, with the flexion side of the forearm as the test site, with an area of ​​3×3 cm 2 The test site should be kept dry, and the sample to be tested should be evenly applied to the flexor side of the arm at about 0.050±0.005g (mL) / time, twice a day, for 7 consecutive days, and the skin reaction should be observed at the same time. If a skin reaction of 3 points or more occurs during this process, it should be decided whether to continue the test according to the specific situation. The skin reaction is observed according to the skin reaction evaluation criteria of the repeated open application test in the "Technical Specifications for Safety of Cosmetics", and the results are recorded, as shown in Table 9.

[0080] Table 9

[0081]

[0082]

[0083] It can be seen that the product of the present invention did not show any adverse reaction in the test, which shows that the product of the present invention has no irritation to human skin and is highly safe.

[0084] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. A freeze-dried essence, characterized in that: It comprises a solution component A and a freeze-dried powder component B; The solution component A includes the following components in parts by weight: 1 part of hyaluronic acid, 3 parts of superoxide dismutase, 0.11 parts of carnosine, 0.6 parts of cinnamaldehyde, 0.6 parts of hexylene glycol, 0.02 parts of Litsea cubeba seed extract and 0.01 parts of Panax notoginseng extract; the freeze-dried powder component B contains the following components in parts by weight: 5 parts of D-mannitol, 0.5 parts of crystalline trehalose, 0.5 parts of soybean polypeptide, 0.1 parts of acetyl hexapeptide-1 and 1 part of fibronectin.

2. The method for preparing the freeze-dried essence according to claim 1, characterized in that: The following steps are involved: (1) Mix the components of freeze-dried powder component B and add water to mix evenly, then freeze-dry and sieve to obtain freeze-dried powder component B; (2) Mix the components of solution component A and add water to mix evenly to obtain solution component A.

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