Preparation method and application of modified bletilla striata polysaccharide for brightening skin

Abstract

The invention belongs to the technical field of natural product extraction and cosmetics, and particularly relates to a method for efficiently extracting bletilla striata polysaccharide by using a betaine deep eutectic solvent (DES), and a bletilla striata polysaccharide-serine transdermal composition prepared by modifying the bletilla striata polysaccharide by using serine. The invention relates to a bletilla striata polysaccharide extraction method, which comprises: using betaine as a hydrogen bond donor, using lactic acid or urea as a hydrogen bond receptor, forming a deep eutectic solvent (DES) according to a molar ratio of 1: 1.5-2, and carrying out ultrasonic-assisted extraction for 10-60 min at a temperature of 50-70 DEG C according to a material-liquid ratio of 1: 10-20 g / mL; the preparation method of the bletilla striata polysaccharide-serine transdermal composition comprises the following steps: dissolving bletilla striata polysaccharide in water at 60-70 DEG C, adding serine, homogenizing and mixing, cooling, adjusting the pH value, and adding a thickening agent; the composition finally achieves the technical effect of brightening the skin through a synergistic effect.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of natural product extraction and cosmetics, and particularly relates to a method for efficiently extracting white plant polysaccharide by using betaine deep eutectic solvent (DES), and a white plant polysaccharide-serine transdermal composition prepared by modifying the white plant polysaccharide with serine. BACKGROUND

[0002] Traditional white plant polysaccharide extraction is usually performed by hot water extraction, which has the problems of low yield (<10%) and high energy consumption. Existing transdermal penetration enhancement technologies (such as chemical penetration enhancers) can irritate the skin. The application solves the above problems by green extraction of betaine DES and the natural moisturizing and transdermal synergistic effect of serine. SUMMARY

[0003] In view of the deficiencies of the prior art, one of the purposes of the application is to provide a method for extracting white plant polysaccharide; another purpose of the application is to provide a white plant polysaccharide-serine transdermal composition; a third purpose of the application is to provide a preparation method of the composition; and a fourth purpose of the application is to provide an application of the composition in the preparation of skin lightening skin care products.

[0004] One of the purposes of the application is achieved by the following technical solutions:

[0005] A method for extracting white plant polysaccharide, wherein betaine is used as a hydrogen bond donor, lactic acid or urea is used as a hydrogen bond acceptor, a deep eutectic solvent (DES) is formed according to a molar ratio of 1:1.5-2, a solid-liquid ratio is 1:10-20 g / mL, and ultrasonic-assisted extraction is performed at 50-70 DEG C for 10-60 minutes.

[0006] Specifically, the molar ratio of betaine to lactic acid in the DES is 1:2.

[0007] Specifically, the ultrasonic power is 150-250 W, and the frequency is 40 kHz.

[0008] The second purpose of the application is achieved by the following technical solutions:

[0009] A white plant polysaccharide-serine transdermal composition, which comprises 0.5-3% of white plant polysaccharide, 0.3-1.5% of serine, 3-8% of a humectant, and 0.05-0.3% of a thickening agent.

[0010] Specifically, the humectant is glycerol or hyaluronic acid, and the thickening agent is xanthan gum or carbomer.

[0011] Specifically, the composition further comprises 0.1-0.5% of a transdermal enhancer, and the transdermal enhancer is lecithin or azone.

[0012] Specifically, the pH value of the composition is 5.0-6.5.

[0013] The third object of the present application is achieved by the following technical solutions:

[0014] A preparation method of a Bletilla striata polysaccharide-serine transdermal composition, comprising: dissolving Bletilla striata polysaccharide in 60-70 DEG C water, adding serine, homogenizing and mixing, adjusting pH after cooling, and adding a thickening agent.

[0015] Specifically, the homogenization speed is 2000-4000 rpm, and the time is 5-10 minutes.

[0016] The fourth object of the present application is achieved by the following technical solutions:

[0017] Application of Bletilla striata polysaccharide in preparing skin lightening skin care products.

[0018] The present application has the following beneficial effects: the DES formed by betaine (hydrogen bond donor) and urea / lactic acid (hydrogen bond acceptor) can destroy the plant cell wall, efficiently extract polysaccharide, and is green and safe; Bletilla striata polysaccharide has the effects of moisturizing, repairing and anti-inflammatory, and is suitable for skin care; serine is a natural moisturizing factor (NMF) component, which can enhance the transdermal property of polysaccharide and promote the hydration of the stratum corneum; and the composition ultimately has the technical effect of skin lightening through synergistic effect. DETAILED DESCRIPTION

[0019] The following will be further described in combination with specific embodiments:

[0020] Example 1-3: DES extraction of Bletilla striata polysaccharide

[0021] Experimental method:

[0022] DES preparation: mix betaine and hydrogen bond acceptor according to the molar ratio in Table 1, and stir at 80 DEG C until transparent.

[0023] Extraction process: mix 10 g of Bletilla striata powder (passed through a 60-mesh sieve) with DES according to the solid-liquid ratio, ultrasonic treatment (conditions are shown in Table 1), centrifuge, and then precipitate the supernatant with 80% ethanol, and freeze-dry to calculate the yield.

[0024] Table 1: Different DES extraction conditions and yields

[0025] Group DES composition (mole ratio) Liquid to solid ratio (g / mL) Temperature (℃) Ultrasonic time (min) Polysaccharide yield (%) 1 Betaine: urea = 1:1.5 1:15 60 30 18.7±0.5 2 Betaine: lactic acid = 1:2 1:20 70 20 21.3±0.6 3* Betaine: glucose = 1:1 1:10 50 40 15.2±0.4

[0026] Note: * is a comparison group

[0027] Conclusion: As shown in Table 1, the extraction method disclosed in the present application can significantly improve the extraction efficiency of Bletilla striata polysaccharide.

[0028] Example 4-6: Preparation of transdermal composition

[0029] Experimental method:

[0030] Formulation: Weigh each component according to Table 2, dissolve in 60℃ deionized water.

[0031] Homogenization: Homogenize at 3000rpm for 5 minutes, and then adjust pH to 5.5-6.0 after cooling.

[0032] Table 2: Transdermal composition formulation (wt%)

[0033] Group Radix bletillae polysaccharide Serine Glycerol Xanthan gum Lecithin Water 4 1.0 0.5 5.0 0.1 0 Balance 5 2.0 1.0 8.0 0.2 0.3 Balance 6* 2.0 0 8.0 0.2 0.3 Balance

[0034] Note: * No serine control group

[0035] Transdermal test experiment

[0036] Experimental method:

[0037] Model establishment: Franz diffusion cell, isolated pig skin (thickness 0.5±0.1mm) as barrier, and PBS (pH 7.4) as receiving liquid.

[0038] Operation: 0.5mL composition (i.e. groups 4-6 recorded in Table 2, all adding 0.1% fluorescent labeled polysaccharide) was added to the supply pool, stirred at 37℃ (600rpm), and sampled at 1, 2, 4, 8, 12h to detect the penetration amount.

[0039] Table 3: Transdermal test results (n=3)

[0040] Group 12h cumulative permeation (pg / cm 2 )]]> Transdermal rate (μg / cm 2 / h) 4 28.4±1.2 2.37±0.10 5 42.7±1.8 3.56±0.15 6 13.6±0.9 1.13±0.08

[0041] Conclusion: According to Table 2 and Table 3, the composition obtained by the present application has higher cumulative penetration amount and scalp rate compared with traditional Bletilla striata whitening products.

[0042] Example 7: Skin lightening effect experiment

[0043] Experimental purpose: To verify the inhibitory effect of the composition containing Bletilla striata polysaccharide and serine on melanin production and the skin lightening effect.

[0044] Experimental method:

[0045] 1, cell experiment (in vitro)

[0046] Model: B16F10 mouse melanoma cells (purchased from Chinese Academy of Sciences Cell Bank).

[0047] Grouping:

[0048] Experimental group: containing the composition of Example 5 (0.1% polysaccharide final concentration)

[0049] Positive control: 0.1% arbutin (known whitening agent)

[0050] Negative control: equal amount of PBS

[0051] Operation:

[0052] Cells were seeded in 96-well plates (1 x 10 4 per well) and incubated at 37°C for 24 h. Melanogenesis was induced by adding medium containing a-MSH (100 nM) with each treatment group. After 48 h incubation, the following were measured: melanin content (NaOH lysis method, 405 nm absorbance); tyrosinase activity (L-DOPA oxidation method, 475 nm absorbance)

[0053] Human trial (clinical)

[0054] Subjects: 30 healthy women (age 25-45 years, with facial pigmentation or dullness).

[0055] Protocol: Apply the composition of Example 5 (0.5 g per time) after cleansing in the morning and evening, for 28 days. The following were measured using a VISIA skin analyzer (Canfield Corporation): melanin index (MI); skin brightness (L* value, CIE Lab colorimetric system)

[0056] Experimental results

[0057] Table 4: In vitro melanin inhibition effect (n = 6)

[0058] Group Melanin content (pg / 10 6 cells) Tyrosinase activity inhibition rate (%) Negative control 35.2±1.8 - Positive control 18.6±0.9 52.3±2.1 Example 5 composition 21.4±1.2 43.7±1.8

[0059] Table 5: Clinical lightening effect (n = 30, day 28)

[0060] Detection index Before use After use Improvement rate (%) Melanin index (MI) 165.3±12.7 138.5±10.2 16.2* Skin brightness (L* value) 58.4±2.1 61.7±1.8 5.7* *P < 0.05 (paired t-test)

[0061] Experimental conclusion

[0062] The composition of Example 5 can significantly inhibit tyrosinase activity (43.7%) and reduce melanin production (the effect is close to that of arbutin). After 28 days of clinical use, the melanin index of the subjects decreased by 16.2%, and the skin brightness increased by 5.7%, confirming its lightening effect.

[0063] Example 8 product (Bletilla striata repair gel)

[0064] The Bletilla striata repair gel was prepared according to the formulation in Table 6.

[0065] Table 6 Formulation of Bletilla striata repair gel

[0066]

[0067]

[0068] Preparation process:

[0069] 1. Clean and sterilize the equipment.

[0070] 2, A phase ahead of soaking evenly, heated to 80-85℃, open slow homogenization 3-4 minutes, cooling began to stir.

[0071] 3, cooling to 60℃ when adding B phase stirring evenly, then add C phase stirring dissolution, continue to cool.

[0072] 4, cooling to 45℃ when adding D phase stirring evenly, can slightly open slow homogenization 1 minute, down to room temperature.

[0073] The above examples and described in the specification is just to illustrate the principles and best embodiments of the present application, without departing from the spirit and scope of the present application, the present application will have various changes and improvements, these changes and improvements are all within the scope of the claimed invention.

Claims

1. A method for extracting Bletilla striata polysaccharide, characterized in that: The low eutectic solvent (DES) is composed of betaine as hydrogen bond donor and lactic acid or urea as hydrogen bond acceptor with a molar ratio of 1:1.5-2, and the material-liquid ratio is 1:10-20 g / mL, and the ultrasonic-assisted extraction is carried out at 50-70 ℃ for 10-60 minutes. ​ 2. The method of claim 1, wherein: The molar ratio of betaine to lactic acid in the DES is 1:

2.

3. The method of claim 1, wherein: The ultrasonic power is 150-250 W, and the frequency is 40 kHz.

4. A bletilla striata polysaccharide-serine transdermal composition, characterized in that: The composition comprises 0.5-3% of the Bletilla striata polysaccharide extracted by the method of any one of claims 1-3, 0.3-1.5% of serine, 3-8% of a humectant, and 0.05-0.3% of a thickening agent.

5. The composition of claim 4, wherein: The humectant is glycerol or hyaluronic acid, and the thickening agent is xanthan gum or carbomer.

6. The composition of claim 4, wherein: The composition further comprises 0.1-0.5% of a transdermal enhancer, which is lecithin or azone.

7. The composition of claim 4, wherein: The pH value of the composition is 5.0-6.

5.

8. A process for the preparation of a composition according to any one of claims 4 to 7, characterised in that: The Bletilla striata polysaccharide is dissolved in water at 60-70 ℃, and the serine is added and homogenized, and after cooling, the pH value is adjusted and the thickening agent is added.

9. The method of claim 8, wherein: The homogenization speed is 2000-4000 rpm, and the time is 5-10 minutes.

10. The use of the Bletilla striata polysaccharide of claim 1 in the preparation of skin lightening skin care products. The formulation is as follows: 。

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