Phalaenopsis fermentation liquor for skin anti-aging and firming, preparation method and application thereof

Abstract

The present application relates to the field of cosmetic technology, especially to IPC A61K8, more particularly to a kind of epiphyllum oxypetalum fermentation liquor for skin anti-aging and firming and its preparation method and application.The preparation method of the epiphyllum oxypetalum fermentation liquor of the present application comprises picking epiphyllum oxypetalum to obtain fermentation substrate;Strain is inoculated on the surface of solid activated culture medium to obtain activated strain;The obtained strain is inoculated into fermentation substrate, and the initial fermentation liquor is obtained by expanding culture;The initial fermentation liquor is treated to obtain a kind of epiphyllum oxypetalum fermentation liquor for skin anti-aging and firming.The epiphyllum oxypetalum fermentation liquor of the present application has good water solubility, is suitable for a variety of cosmetic formulations, and the preparation method in the present application is simple, safe and green, and is suitable for industrial production.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of cosmetics, in particular to IPC A61K8, and more particularly to a kind of epiphyllum oxycnodon fermentation liquor for skin anti-aging and firming and its preparation method and application. BACKGROUND

[0002] With age and environmental factors, the appearance, physical properties and physiological functions of human skin change. In the aging skin, the epidermal cell renewal slows down, the barrier function weakens, and the repair ability of the epidermis after injury weakens, leading to dry skin, desquamation, wrinkles and the like; In the dermis layer of the aging skin, the number of fibroblasts gradually decreases, the ability to synthesize collagen and elastin decreases, and the deposition of damaged proteins increases, the most direct manifestation is wrinkles, relaxation and pigmentation.

[0003] Epiphyllum oxycnodon, also known as jionghua and yuexia meiren, belongs to the plant of cactus family Epiphyllum, its flower and stem are used as medicine, which has the effects of clearing lung and relieving cough, cooling blood and stopping bleeding, and nourishing heart and tranquilizing mind. Modern research shows that the active ingredients of epiphyllum oxycnodon are flavonoids, among which the aglycone is mainly quercetin, isorhamnetin and kaempferol.

[0004] There are many methods for extracting bioactive ingredients from plants. In recent years, the development trend of microbial fermentation technology in the cosmetics industry has risen sharply. The fermentation product filtrate obtained by fermentation technology contains rich amino acids, polypeptides, minerals and various vitamins, which are important components of human body and play an important role in improving skin conditions.

[0005] The existing patent CN113425622A discloses an extraction with multiple plant compositions, in which 70% ethanol reflux extraction is used in multiple steps, the extraction steps are complex, the extraction rate is low, the loss of active ingredients is large, and the organic solvent residue in the obtained extract is easy to cause harm to the skin.

[0006] Therefore, it is necessary to develop an epiphyllum oxycnodon fermentation liquor with high active ingredient content, high extraction rate, simple steps and less organic solvent residue. SUMMARY

[0007] In order to solve the problems in the prior art, the first aspect of the present application provides a preparation method of epiphyllum oxycnodon fermentation liquor for skin anti-aging and firming, comprising the following steps:

[0008] S1: picking epiphyllum oxycnodon to obtain fermentation substrate;

[0009] S2: inoculating the strain on the surface of solid activated culture medium to obtain activated strain;

[0010] S3: inoculating the strain obtained in S2 into the fermentation substrate in S1 to enlarge the culture to obtain initial fermentation liquor;

[0011] S4: processing the initial fermentation liquor to obtain a Passiflora edulis fermentation liquor for skin anti-aging and tightening.

[0012] Preferably, the preparation method of the Passiflora edulis fermentation liquor for skin anti-aging and tightening comprises the following specific steps:

[0013] S1, taking fresh Passiflora edulis, low-temperature drying, crushing, sieving, to obtain Passiflora edulis powder, adding pure water to mix uniformly, sterilizing at 121°C for 20-50 min to obtain a fermentation substrate;

[0014] S2, inoculating the strain on the surface of a solid activated culture medium, activating the culture at 25-38°C for 12-36 h to obtain an activated strain;

[0015] S3, inoculating the strain obtained in S2 into the fermentation substrate in S1 at an inoculation amount of 1-5%, and performing shake culture at 35-45°C for 36-72 h to obtain an initial fermentation liquor;

[0016] S4, sequentially performing sterilization, cell wall breaking, centrifugation, and filtration to collect the filtrate, thereby obtaining a Passiflora edulis fermentation liquor for skin anti-aging and tightening.

[0017] Preferably, the temperature of the low-temperature drying is 30-40°C.

[0018] Preferably, the particle size of the Passiflora edulis powder in S1 is 60-200 mesh.

[0019] Preferably, the mass ratio of the Passiflora edulis powder to pure water in S1 is 1:10-30 g / g; further preferably, 1:20.

[0020] Preferably, the strain in S2 comprises one or more of Bifidobacterium and lactic acid bacteria; further preferably, Bifidobacterium.

[0021] Preferably, the inoculation amount of the strain in S2 is 0.5-3%; further preferably, 3%.

[0022] Preferably, the Bifidobacterium is purchased from the China General Microbiological Culture Collection Center, CGMCC 1.2190.

[0023] Preferably, the solid activated culture medium is BBL liquid medium.

[0024] Preferably, the solid activated culture medium is purchased from Shanghai Yage Biological Technology Co., Ltd., with the product number HB8777.

[0025] Preferably, the sterilization in S4 is performed at 121°C for 20-60 min, further preferably, 30 min.

[0026] Preferably, the instrument for the cell wall breaking treatment is a high pressure homogenizer.

[0027] Preferably, the temperature for the cell wall breaking treatment is 70-90℃, the pressure is 400-600bar, the time is 4-6min, the interval is 4-6min, and the number of repetitions is 3-5.

[0028] Preferably, the centrifugation is performed at a speed of 1500-10000rpm for 10-30min, and more preferably, at a speed of 5000rpm for 20min.

[0029] Preferably, the filtration is performed using a plate-and-frame filter membrane with a pore size of 0.2-0.8μm.

[0030] In the present application, the epiphyllum treatment is used as a fermentation substrate, and is fermented with specific strains of bifidobacterium or lactic acid bacteria. The method is simple, safe, and green, and is suitable for industrial production. The present inventors first used epiphyllum as a fermentation substrate, inoculated bifidobacterium or lactic acid bacteria, and unexpectedly found that the epiphyllum fermentation broth after cultivation and treatment had anti-wrinkle and firming effects. The present inventors speculate that bifidobacterium uses epiphyllum as a substrate, releases various proteases and cellulases during fermentation, destroys the cell walls of epiphyllum cells, decomposes the proteins and sugars in epiphyllum, and releases active substances in epiphyllum. On the other hand, bifidobacterium grows and reproduces during the absorption of nutrients, and produces secondary metabolites. It is possible that the secondary metabolites of bifidobacterium and the active substances released from epiphyllum act together to improve the anti-wrinkle and firming effects of the epiphyllum fermentation broth. If the active ingredients in epiphyllum are destroyed in advance, the fermentation performance of bifidobacterium will be weakened, the functional ingredients in the fermentation broth will be reduced, and the anti-wrinkle and firming effects will be weakened.

[0031] In the present application, epiphyllum is used as a fermentation substrate, and the fermentation time is precisely controlled to ensure the types and contents of bioactive ingredients and guarantee the anti-aging and firming effects. The present inventors speculate that precise control of the fermentation time can maximize the destruction of the cell walls of epiphyllum under the action of the enzymes released by bifidobacterium, and release a large amount of active ingredients. At the same time, bifidobacterium is in a rapid reproduction period, and produces a large amount of secondary metabolites. If the fermentation time is too short, the amount of active substances released from epiphyllum is not enough. If the fermentation time is too long, bifidobacterium may die due to lack of nutrients. These will affect the efficacy of the epiphyllum fermentation broth.

[0032] The second aspect of the present application provides an epiphyllum fermentation broth prepared by the preparation method for the skin anti-aging and firming epiphyllum fermentation broth.

[0033] The third aspect of the present application provides an application of the phalaenopsis fermenting liquid prepared by the preparation method of the skin anti-aging and firming phalaenopsis fermenting liquid, which is applied to cosmetics; the dosage form of the cosmetic includes water agent, emulsion, cream, mask, and foundation.

[0034] The mass percentage of the phalaenopsis fermenting liquid in the cosmetic is 0.1-10%.

[0035] Beneficial effects

[0036] 1. In the present application, the phalaenopsis processing product is used as a fermentation substrate, and specific strains, bifidobacterium or lactic acid bacteria are fermented to obtain the phalaenopsis fermenting liquid with anti-wrinkle and firming effects.

[0037] 2. In the present application, phalaenopsis is used as a fermentation substrate, and the fermentation time is accurately controlled to ensure the types and contents of bioactive ingredients and guarantee the anti-aging and firming effects.

[0038] 3. In the present application, the high-pressure homogenizer is used for wall breaking treatment of the strains, which further improves the dissolution of the effective components and is simple, safe and efficient.

[0039] 4. The phalaenopsis fermenting liquid prepared in the present application has good water solubility and is suitable for application in various cosmetic formulations.

[0040] 5. In the present application, unlike the simple heating water extraction of general plant extract, high-temperature sterilization is directly performed, which highly retains the active ingredients in the phalaenopsis fermentation substrate, so that the obtained phalaenopsis fermenting liquid has good effects of promoting the growth of collagen and elastin, thereby improving the anti-wrinkle and firming effects of the fermenting liquid. BRIEF DESCRIPTION OF DRAWINGS

[0041] Figure 1 It is a Collagen I protein content result summary chart in the anti-wrinkle and firming effect test of the present application example 1 and comparative examples 1-4;

[0042] Figure 2 It is an elastin content result summary chart in the anti-wrinkle and firming effect test of the present application example 1 and comparative examples 1-4;

[0043] Figure 3 It is a test result chart of the skin wrinkle improvement rate of the present application example 1;

[0044] Figure 4 It is a test result chart of the skin elasticity improvement rate of the present application example 1;

[0045] Figure 5 It is a test result chart of the eye corner wrinkle improvement rate of the present application example 1. DETAILED DESCRIPTION

[0046] Example 1

[0047] The first aspect of the embodiment provides a preparation method of a cactus fermentation liquid for skin anti-aging and firming, comprising the following steps:

[0048] S1, take 200g of freshly picked cactus (Epiphyllum oxypetalum), dry in a 30℃ oven, crush, pass through a 100 mesh sieve, add pure water and disperse uniformly, control the mass ratio to be 1:20g / g, sterilize at 121℃ for 30min to obtain a fermentation substrate;

[0049] S2, inoculate Bifidobacterium on the surface of a solid activated culture medium, activate at 37℃ for 18h to obtain an activated strain;

[0050] S3, inoculate the strain obtained in S2 into the fermentation substrate in S1 at an inoculation amount of 3%, expand the culture, and ferment at 37℃ on a shaking table for 48h to obtain an initial fermentation liquid;

[0051] S4, sterilize the initial fermentation liquid at 121℃ for 30min, perform cell wall breaking treatment using a high-pressure homogenizer, temperature 75℃, pressure 500bar, cell wall breaking for 5min, stop for 5min, repeat for 3 times, centrifuge at 5000rpm for 20min, collect the filtrate by filtering with a plate and frame filter membrane, and the pore size of the filter membrane is 0.2μm, thereby obtaining a cactus fermentation liquid for skin anti-aging and firming.

[0052] The cactus is purchased from Xinfeng E-commerce Company in Nancheng County.

[0053] The Bifidobacterium is purchased from China General Microbiological Culture Collection Center, CGMCC 1.2190.

[0054] The solid activated culture medium is purchased from Shanghai Yaji Biological Technology Co., Ltd., item number: HB8777.

[0055] The embodiment provides an application of the cactus fermentation liquid prepared by the preparation method of the cactus fermentation liquid for skin anti-aging and firming, and the cactus fermentation liquid is applied to cosmetics; the dosage form of the cosmetics is emulsion.

[0056] The mass percentage of the cactus fermentation liquid in the cosmetics is 2%.

[0057] Comparative Example 1

[0058] The comparative example provides a preparation method of a cactus fermentation liquid for skin anti-aging and firming, comprising the following steps:

[0059] Take 200g of fresh picked orchid, oven drying at 30℃, crushing, passing through 100 mesh sieve, adding pure water evenly dispersed, control mass ratio of 1:20g / g, heated reflux at 75℃ for 1h, heated reflux extraction 2 times, filtration, combined filtrate, to obtain a kind of orchid fermentation broth.

[0060] The orchid is purchased from Xinfeng E-commerce Company in Nancheng County.

[0061] Comparative Example 2

[0062] The first aspect of the present comparative example provides a preparation method of an orchid fermentation broth for skin anti-aging and tightening, comprising the following steps:

[0063] S1, take 200g of fresh picked orchid, oven drying at 30℃, crushing, passing through 100 mesh sieve, adding pure water evenly dispersed, control mass ratio of 1:20g / g, heated reflux at 75℃ for 1h, heated reflux extraction 2 times, filtration, combined filtrate, standby;

[0064] S2, inoculate Bifidobacterium on the surface of solid activated culture medium, activate culture at 37℃ for 18h, obtain activated strain;

[0065] S3, inoculate the strain obtained in S2 into Bifidobacterium culture medium (BBL liquid culture medium) with an inoculation amount of 3%, expand culture at 37℃ on a shaking table for 48h, sterilize at 121℃ for 30min, to obtain initial fermentation broth;

[0066] S4, add the filtrate obtained in S1 to the fermentation broth in S3, use high-pressure homogenizer for cell wall breaking treatment, temperature 75℃, pressure 500bar, cell wall breaking for 5min, stop interval for 5min, repeat for 3 times, centrifuge at 5000rpm for 20min, collect the filtrate by filtering with plate and frame filter membrane, the pore size of the filter membrane is 0.2μm, to obtain an orchid fermentation broth.

[0067] The orchid is purchased from Xinfeng E-commerce Company in Nancheng County.

[0068] The Bifidobacterium is purchased from China General Microbiological Culture Collection Center, CGMCC 1.2190.

[0069] The solid activated culture medium is purchased from Shanghai Yaji Biological Technology Co., Ltd., item number: HB8777.

[0070] Comparative Example 3

[0071] S1, take 200g of fresh picked orchid, oven drying at 30℃, crushing, passing through 100 mesh sieve, adding pure water evenly dispersed, control mass ratio of 1:20g / g, sterilize at 121℃ for 30min, to obtain fermentation substrate;

[0072] S2, inoculate the Bifidobacterium on the surface of solid activation medium, activate culture at 37℃ for 18h, and obtain activated strains;

[0073] S3, inoculate the strains obtained in S2 into the fermentation medium in S1 at an inoculation amount of 3%, expand culture, and perform fermentation culture at 37℃ for 24h on a shaking table to obtain an initial fermentation liquor;

[0074] S4, sterilize the initial fermentation liquor at 121℃ for 30min, perform cell wall breaking treatment by using a high-pressure homogenizer, the temperature is 75℃, the pressure is 500bar, the cell wall breaking time is 5min, the interval is 5min, repeat for 3 times, centrifuge at a speed of 5000rpm for 20min, filter the filtrate by using a plate and frame filter membrane, the pore size of the filter membrane is 0.2μm, and thus a Passiflora edulis fermentation liquor for skin anti-aging and tightening is obtained.

[0075] The present comparative example provides an application of a Passiflora edulis fermentation liquor prepared by a preparation method of a Passiflora edulis fermentation liquor for skin anti-aging and tightening, which is applied to a cosmetic; the dosage form of the cosmetic is emulsion.

[0076] The mass percentage of the Passiflora edulis fermentation liquor in the cosmetic is 2%.

[0077] The Passiflora edulis is purchased from Xinfeng E-commerce Company in Nancheng County.

[0078] The Bifidobacterium is purchased from China General Microbiological Culture Collection Center, CGMCC 1.2190.

[0079] The solid activation medium is purchased from Shanghai Yaji Biological Technology Co., Ltd., and the article number is HB8777.

[0080] Comparative Example 4

[0081] S1, take 200g of freshly picked Passiflora edulis, dry in a 30℃ oven, crush, pass through a 100-mesh sieve, uniformly disperse by adding pure water, control the mass ratio to be 1:20g / g, sterilize at 121℃ for 30min, and obtain a fermentation medium;

[0082] S2, inoculate the Bifidobacterium on the surface of solid activation medium, activate culture at 37℃ for 18h, and obtain activated strains;

[0083] S3, inoculate the strains obtained in S2 into the fermentation medium in S1 at an inoculation amount of 3%, expand culture, and perform fermentation culture at 37℃ for 48h on a shaking table to obtain an initial fermentation liquor;

[0084] S4, sterilize the initial fermentation liquor at 121℃ for 30min, centrifuge at a speed of 5000rpm for 20min, filter the filtrate by using a plate and frame filter membrane, the pore size of the filter membrane is 0.2μm, and thus a Passiflora edulis fermentation liquor for skin anti-aging and tightening is obtained.

[0085] The present comparative example provides an application of a phalaenopsis flower fermentation liquid prepared by a preparation method of a skin anti-aging and firming phalaenopsis flower fermentation liquid, which is applied to a cosmetic; the dosage form of the cosmetic is an emulsion.

[0086] The mass percentage of the phalaenopsis flower fermentation liquid in the cosmetic is 2%.

[0087] The phalaenopsis flower is purchased from Xinfeng E-commerce Company in Nancheng County.

[0088] The Bifidobacterium is purchased from China General Microbiological Culture Collection Center, CGMCC 1.2190.

[0089] The solid activated culture medium is purchased from Shanghai Yaji Biological Technology Co., Ltd., and the item number is HB8777.

[0090] Performance test

[0091] 1. Fibroblast experiment

[0092] In test example 1, the changes in the contents of Collagen I and Elastin in the phalaenopsis flower fermentation liquids prepared in comparative examples 1, 2, 3 and 4 were tested to analyze and evaluate the anti-wrinkle and firming effects.

[0093] Collagen I is Collagen I, which accounts for about 80% of the dermis of the skin, makes the skin full and plump, and an increase in the content can achieve the effect of resisting wrinkles. Elastin is the main protein that constitutes elastic fibers, and an increase in the content can improve the elasticity of the skin. In this performance test experiment, fibroblasts were used as the research model, and an in vitro photoaging model was established by UVA irradiation. By detecting the changes in the contents of Collagen I and Elastin, the anti-wrinkle and firming effects of the test substance were analyzed.

[0094] Test method: see Table 1 for test scheme.

[0095] Table 1

[0096]

[0097]

[0098] Cells were inoculated into a 6-well plate at an inoculation density of 2x10 5 According to the test scheme in Table 1, when the plating rate of the cells in the 6-well plate reached 40%-60%, the cells were dosed according to the experimental grouping, 2 mL was added to each well, 3 replicate wells were set for each group, and the cells were incubated in an incubator (37°C, 5% CO2) for 24 h. According to the grouping, the cells were irradiated with 30 J / cm 2UVA irradiation, and placed in an incubator (37℃, 5% CO2) for continuous culture for 24 h. The cell culture supernatant was collected and detected according to the ELISA kit instructions.

[0099] The test results are shown in Figure 1 、 Figure 2 . Figure 1 、 Figure 2 The significance is represented by # compared with the blank control group, and ###, ##, # represent 1%, 5%, and 10% significance levels, respectively. The significance is represented by ***, **, * compared with the negative control group, and 1%, 5%, and 10% significance levels, respectively.

[0100] It can be seen from Figure 1 and Figure 2 that:

[0101] Compared with the blank control group, the Collagen I protein and elastin content of the negative control group were significantly decreased, indicating that the test stimulation condition was effective.

[0102] Compared with the negative control group, the Collagen I protein and elastin content of the positive control group were significantly increased, indicating that the positive control of this test was effective.

[0103] Compared with the negative control group, the Collagen I protein content of Comparative Examples 2, 3, 4, and Example 1 was significantly increased, and the improvement effect was in the order of Example 1 > Comparative Example 3 > Comparative Example 2 > Comparative Example 4 > Comparative Example 1.

[0104] Compared with the negative control group, the elastin content of Comparative Examples 2, 3, 4, and Example 1 was significantly increased, and the improvement effect was in the order of Example 1 > Comparative Example 3 > Comparative Example 2 > Comparative Example 4 > Comparative Example 1.

[0105] 2. Human test

[0106] Thirty-three healthy volunteers were selected, regardless of gender, and aged 30-45 years. An emulsion containing 2% of the Passiflora incarnata ferment prepared in Example 1 was used, and the whole face was used, once in the morning and once in the evening, continuously for 4 weeks. The skin wrinkles were measured using Antera3D, and the skin elasticity and tightness were measured using the Cutometer MPA580 skin elasticity tester. After 2 weeks and 4 weeks, the overall size of the skin wrinkles of the 33 volunteers was measured, and the average value was calculated. After 2 weeks and 4 weeks, the skin rebound ratio of the 33 volunteers was measured, and the average value was calculated. After 2 weeks and 4 weeks, the skin tightness of the 33 volunteers was measured, and the average value was calculated. The results are shown in Figure 3 、 Figure 4 、 Figure 5 . The emulsion formula includes:

[0107] Ingredient Amount (%) Hydrogenated poly(C6-14-alkene) 4 Glycerin 3.071 1,2-Pentanediol 1.5 Glyceryl stearate 0.6 Stearyl alcohol 0.505 1,2-Hexanediol 0.501 Cetyl alcohol 0.492 Cyclopentasiloxane 0.425 Octoxyglycerin 0.3 PEG-100 stearate 0.3 Tromethamine 0.18 Carbomer 0.12 Butyrospermum parkii (shea) butter 0.1 (Fragrance) 0.08 Dimethicone 0.075 Acrylates / C10-30 alkyl acrylate crosspolymer 0.06 Ethylhexylglycerin 0.049975 Sodium phytate 0.02 Epiphyllum cactus fermentation broth prepared in Example 1 2 Water Balance

[0108] From Figure 3 , Figure 4 and Figure 5 it follows that:

[0109] Compared with before use, the overall size of skin wrinkles after 2w, 4w of use was significantly reduced by 10.05%, 24.02% respectively.

[0110] Compared with before use, the skin rebound ratio (the closer the R2 value is to 1, the better the skin elasticity) after 2w, 4w of use was significantly increased by 23.16%, 37.37% respectively.

[0111] Compared with before use, the eye corner wrinkle tightness (the smaller the F4, the better the skin tightness) after 2w, 4w of use was significantly decreased by 19.99%, 30.44% respectively.

Claims

1. A method for preparing a Passiflora edulis fermentate for skin anti-aging firming, characterized by, The preparation method comprises the following specific steps: S1, taking fresh picked orchid, low temperature drying, crushing, sieving, getting orchid powder, adding pure water, mixing evenly, sterilizing at 121℃ for 20-50min, getting fermentation substrate; S2, inoculating the strain on the surface of solid activated culture medium, activating at 25-38℃ for 12-36h, getting activated strain; S3, expanding culture of the strain obtained in S2, adding to the fermentation substrate of S1 with 1-5% inoculation amount, shaking culture at 35-45℃ for 36-72h, getting initial fermentation liquor; S4, the initial fermentation liquor is subjected to sterilization, wall breaking treatment, centrifugation, filtration to collect filtrate, getting an orchid fermentation liquor for skin anti-aging and tightening; The bifidobacterium is purchased from China General Microbiological Culture Collection Center, CGMCC 1.2190.

2. The method of claim 1, wherein the preparation of the Epiphyllum cactus fermentation broth for skin anti-aging tightening is characterized by, The particle size of the orchid powder in S1 is 60-200 meshes.

3. The method of claim 1, wherein the preparation of the Epiphyllum cactus fermentation broth for skin anti-aging tightening is characterized by, The inoculation amount of the strain in S2 is 0.5-3%.

4. The method of claim 1, wherein the preparation of the Epiphyllum cactus fermentation broth for skin anti-aging tightening is characterized by, The sterilization treatment in S4 is sterilization at 121℃ for 20-60min.

5. The method of claim 1, wherein the preparation of the Epiphyllum cactus fermentation broth for skin anti-aging tightening is characterized by, The wall breaking treatment instrument is high pressure homogenizer.

6. The method of claim 1, wherein the preparation of the Epiphyllum cactus fermentation broth for skin anti-aging tightening is characterized by, The wall breaking treatment temperature is 70-90℃, the pressure is 400-600bar, the time is 4-6min, the interval is 4-6min, and the repetition number is 3-5 times.

7. An orchid fermentation liquor prepared by the preparation method of the orchid fermentation liquor for skin anti-aging and tightening according to any one of claims 1-6.

8. The Epiphyllum Fermented Liquid for skin anti-aging and firming according to the preparation method of claim 7, characterized in that, It is applied to cosmetics; the dosage form of the cosmetic comprises water agent, emulsion, cream, mask, foundation.

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