Radix pseudostellariae cyclic peptide extract and preparation method thereof

Abstract

The invention relates to a radix pseudostellariae cyclic peptide extract and a preparation method thereof, and belongs to the technical field of natural active ingredient extraction. The preparation method of the heterophylly falsestarwort root cyclopeptide extract comprises the following steps: crushing heterophylly falsestarwort root to obtain heterophylly falsestarwort root powder, soaking the heterophylly falsestarwort root powder in water to obtain a heterophylly falsestarwort root soaking solution, inoculating a complex microbial inoculant into the heterophylly falsestarwort root soaking solution, carrying out fermentation culture to obtain a fermentation product, and then extracting to obtain a heterophylly falsestarwort root cyclopeptide extracting solution, performing freeze drying to obtain a radix pseudostellariae cyclic peptide extract; the complex microbial inoculant is a complex microbial inoculant of bacillus subtilis and clostridium butyricum, and the mass ratio of the bacillus subtilis to the clostridium butyricum in the complex microbial inoculant is 1: (1-3). The heterophylly falsestarwort root cyclic peptide extract is extracted through fermentation culture of the complex microbial inoculant of the bacillus subtilis and the clostridium butyricum, the method is safe and environmentally friendly, use of organic solvents is avoided, and the concept of green and environmental protection is met.

Description

Technical Field

[0001] The invention belongs to the technical field of natural active ingredient extraction, and particularly relates to a Pseudostellaria heterophylla cyclic peptide extract and a preparation method thereof. Background Art

[0002] Radix Pseudostellariae belongs to the plant Panax notoginseng of the Caryophyllaceae family. It is a traditional Chinese medicine in my country. It tastes sweet, slightly bitter, and flat. It enters the spleen and lung meridians. It has the effects of invigorating qi and strengthening the spleen, promoting the production of body fluids and moistening the lungs. It is used for spleen deficiency, fatigue, loss of appetite, weakness after illness, insufficient qi and yin, spontaneous sweating and thirst, and dry cough due to lung dryness. Existing studies have shown that Radix Pseudostellariae contains polysaccharides, cyclic peptides, saponins and other ingredients, which have anti-fatigue, anti-hypoxia, anti-wrinkle, anti-aging effects, among which cyclic peptides are its characteristic ingredients. In order to promote the development of the Radix Pseudostellariae industry and develop related functional products and skin care products of Radix Pseudostellariae cyclic peptide ingredients, how to effectively obtain extracts containing Radix Pseudostellariae cyclic peptide ingredients has become one of the problems to be solved urgently. The extraction methods of Radix Pseudostellariae cyclic peptide extracts in the existing technology are generally alcohol extraction, silica gel column chromatography, etc., which have high production costs and use a large amount of organic solvents, which are not in line with the concept of green environmental protection. Summary of the Invention

[0003] The first object of the present invention is to provide a method for preparing Pseudostellaria baicalensis cyclic peptide extracts to solve the technical problem of high production cost when extracting Pseudostellaria baicalensis cyclic peptide extracts by alcohol extraction and silica gel column chromatography.

[0004] The second object of the present invention is to provide a Pseudostellaria baicalensis cyclic peptide extract.

[0005] In order to achieve the above objectives, the technical solution adopted by the present invention is: A method for preparing a Pseudostellaria chinensis cyclic peptide extract comprises the following steps: crushing Pseudostellaria chinensis to obtain Pseudostellaria chinensis powder, soaking the Pseudostellaria chinensis powder in water to obtain a Pseudostellaria chinensis soaking liquid, inoculating a composite bacterial agent into the Pseudostellaria chinensis soaking liquid for fermentation and culturing to obtain a fermentation product, subsequently extracting to obtain a Pseudostellaria chinensis cyclic peptide extract, and freeze-drying to obtain the Pseudostellaria chinensis cyclic peptide extract; the composite bacterial agent is a composite bacterial agent of Bacillus subtilis and Clostridium butyricum, and the mass ratio of Bacillus subtilis to Clostridium butyricum in the composite bacterial agent is 1:1-3.

[0006] Furthermore, the inoculation amount of the composite bacterial agent is 10-15%.

[0007] Furthermore, the fermentation temperature is 33-37° C., the fermentation time is 30-50 h, and the pH of the fermentation is 6-8.

[0008] Furthermore, the extraction method is: extracting at 30-40°C on a constant temperature shaker at a speed of 200-300 rpm for 0.5-1 h, followed by centrifugation at 5000-6000 r / min for 10-30 min.

[0009] Furthermore, before extraction, the concentration of the fermentation product is diluted to 30-50 g / L.

[0010] Furthermore, the mass volume ratio of the Pseudostellariae Radix powder to water is 1 g: 10-15 mL, and the soaking time is 30-90 min.

[0011] Furthermore, the mesh size of the Radix Pseudostellariae powder is 20 to 30 meshes.

[0012] Furthermore, the freeze-drying method is: pre-freezing at normal pressure and -30 to -50°C for 30 to 60 minutes, and then drying at -70 to -80°C in vacuum for 30 to 60 minutes.

[0013] A Pseudostellaria cylindrica cyclic peptide extract is prepared by adopting the above-mentioned preparation method of the Pseudostellaria cylindrica cyclic peptide extract.

[0014] Beneficial effects of the present invention: The invention extracts the Pseudostellaria heterophylla cyclic peptide extract by fermentation culture of a composite bacterial agent of Bacillus subtilis and Clostridium butyricum, is safe and environmentally friendly, avoids the use of organic solvents, and complies with the concept of green environmental protection.

[0015] The present invention combines Bacillus subtilis and Clostridium butyricum, and the extraction rate of the Pseudostellariae cyclic peptide extract is high. The contents of Pseudostellariae cyclic peptide A and Pseudostellariae cyclic peptide B in the extracted Pseudostellariae cyclic peptide extract are significantly increased. The Pseudostellariae cyclic peptide extract prepared by the present invention can be used in anti-wrinkle skin care products.

[0016] It can be seen from Comparative Examples 1 and 2 of the present invention that the extraction rate of the Pseudostellariae cyclic peptide extract obtained by fermentation with a single bacterium is low, and the contents of Pseudostellariae cyclic peptide A and Pseudostellariae cyclic peptide B in the extracted Pseudostellariae cyclic peptide extract are low. It can be seen from Comparative Example 4 of the present invention that the extraction rate and the contents of Pseudostellariae cyclic peptide A and Pseudostellariae cyclic peptide B obtained by fermentation with a composite bacterial agent of Bacillus subtilis and Lactobacillus are also low. The effect of the present invention can not be achieved by using a composite bacterial agent of any two bacteria. In the present invention, Bacillus subtilis and Clostridium butyricum have a compounding effect. DETAILED DESCRIPTION

[0017] The present invention will be further described below in conjunction with the embodiments of the present invention.

[0018] The deposit number of Clostridium butyricum is CCTCC NO: M2024807.

[0019] The deposit number of Bacillus subtilis is CGMCC No.29125.

[0020] The preservation number of Lactobacillus is CGMCC NO.23158. Example 1

[0021] The preparation method of the Pseudostellariae cyclic peptide extract of Example 1 is as follows: 1 g of Radix Pseudostellariae was crushed to obtain Radix Pseudostellariae powder with a mesh size of 20 mesh. 1 g of Radix Pseudostellariae powder was soaked in 10 mL of water for 30 min to obtain Radix Pseudostellariae soaking solution, which was sterilized at 121°C. Subsequently, the composite bacterial agent was inoculated into the Radix Pseudostellariae soaking solution at an inoculum size of 12% (v / v). The mixture was fermented in a fermentation chamber at 35°C for 50 h. The pH of the fermentation culture was 7. After the fermentation culture was completed, the mixture was inactivated to obtain the fermentation product. The concentration of the fermentation product after fermentation culture was adjusted to 30 g / L with ultrapure water. The mixture was extracted in a constant temperature shaker at 30°C and 200 rpm for 0.5 h, and then centrifuged at 5000 r / min for 15 min. The supernatant was obtained as the Radix Pseudostellariae cyclic peptide extract. The Radix Pseudostellariae cyclic peptide extract was pre-frozen at -30°C under normal pressure for 30 min, and then dried at -70°C under vacuum for 60 min to obtain the Radix Pseudostellariae cyclic peptide extract. The mass ratio of Bacillus subtilis to Clostridium butyricum in the composite bacterial agent is 1: 2. The Pseudostellaria heterophylla of Example 1 is produced in Guizhou. Example 2

[0022] The preparation method of the Pseudostellariae cyclic peptide extract of Example 2 is as follows: 3 g of Radix Pseudostellariae was crushed to obtain Radix Pseudostellariae powder with a mesh size of 20 mesh. 3 g of Radix Pseudostellariae powder was soaked in 45 mL of water for 90 min to obtain Radix Pseudostellariae soaking solution. The Radix Pseudostellariae soaking solution was sterilized at 121°C, and then the composite bacterial agent was inoculated into the Radix Pseudostellariae soaking solution at an inoculum size of 15% (v / v). The mixture was fermented in a fermentation chamber at 37°C for 45 h. The pH of the fermentation culture was 8. After the fermentation culture was completed, the mixture was inactivated to obtain the fermentation product. The concentration of the fermentation product after fermentation culture was adjusted to 50 g / L with ultrapure water, and the mixture was extracted in a constant temperature shaker at 40°C and 200 rpm for 0.5 h. The mixture was then centrifuged at 6000 r / min for 15 min, and the supernatant was obtained, which was the Radix Pseudostellariae cyclic peptide extract. The Radix Pseudostellariae cyclic peptide extract was pre-frozen at -50°C at normal pressure for 30 min, and then dried at -80°C in a vacuum for 30 min to obtain the Radix Pseudostellariae cyclic peptide extract. The mass ratio of Bacillus subtilis to Clostridium butyricum in the composite bacterial agent is 1: 1. The Pseudostellaria heterophylla of Example 2 is produced in Guizhou. Example 3

[0023] The preparation method of the Pseudostellariae cyclic peptide extract of Example 3 is as follows: 100 g of Radix Pseudostellariae was crushed to obtain Radix Pseudostellariae powder, the mesh number of Radix Pseudostellariae powder was 30 mesh, 100 g of Radix Pseudostellariae powder was soaked in 1000 mL of water for 60 min to obtain Radix Pseudostellariae soaking solution, the Radix Pseudostellariae soaking solution was sterilized at 121 °C, and then the composite bacterial agent was inoculated into the Radix Pseudostellariae soaking solution at an inoculum amount of 10% (v / v), and fermented in a fermentation box at 33 °C for 50 h, the pH of the fermentation culture was 7, and the fermentation product was inactivated after the fermentation culture was completed. The concentration of the fermentation product after fermentation culture was adjusted to 40 g / L with ultrapure water, and extracted on a constant temperature shaker at 30 °C and 300 rpm for 0.5 h, and then centrifuged at 6000 r / min for 30 min, and the supernatant was taken, namely the Radix Pseudostellariae cyclic peptide extract, and the Radix Pseudostellariae cyclic peptide extract was pre-frozen at normal pressure and -40 °C for 60 min, and then dried at -80 °C in vacuum for 60 min, namely, the Pseudostellaria heterophylla cyclic peptide extract. The mass ratio of Bacillus subtilis and Clostridium butyricum in the composite bacterial agent is 1:3. The Pseudostellaria heterophylla of Example 3 is produced in Guizhou.

[0024] Examples 4 to 6 The preparation methods of the Pseudostellariae cyclic peptide extracts of Examples 4 to 6 are substantially the same as those of Examples 1 to 3. The difference between the preparation method of the Pseudostellariae cyclic peptide extract of Example 4 and Example 1 is that the Pseudostellariae cyclic peptide extract of Example 4 is produced in Nanjing.

[0025] The difference between the preparation method of the Pseudostellariae Radix cyclopeptide extract of Example 5 and that of Example 2 is that the Pseudostellariae Radix in Example 5 is produced in Nanjing.

[0026] The difference between the preparation method of the Pseudostellariae Radix cyclic peptide extract of Example 6 and that of Example 3 is that the Pseudostellariae Radix in Example 6 is produced in Nanjing.

[0027] Comparative Example 1 The preparation method of the Pseudostellariae cyclic peptide extract of Comparative Example 1 is different from that of Example 1 in that the composite bacterial agent in Comparative Example 1 is replaced by Bacillus subtilis, and the inoculation rate of Bacillus subtilis is 15% (v / v).

[0028] Comparative Example 2 The preparation method of the Pseudostellariae cyclic peptide extract of Comparative Example 2 is different from that of Example 1 in that the composite bacterial agent in Comparative Example 2 is replaced by Clostridium butyricum, and the inoculation rate of Clostridium butyricum is 15% (v / v).

[0029] Comparative Example 3 The difference between the preparation method of the Pseudostellariae cyclic peptide extract of Comparative Example 3 and that of Example 1 is that the mass ratio of Bacillus subtilis to Clostridium butyricum in Comparative Example 3 is 1:4.

[0030] Comparative Example 4 The difference between the preparation method of the Pseudostellariae cyclic peptide extract of Comparative Example 4 and that of Example 1 is that the composite bacterial agent in Comparative Example 4 is Bacillus subtilis and Lactobacillus.

[0031] Comparative Examples 5 to 8 The preparation methods of the Pseudostellariae cyclic peptide extracts of Comparative Examples 5 to 8 are roughly the same as the preparation methods of the Pseudostellariae cyclic peptide extracts of Comparative Examples 1 to 4. The difference between the preparation method of the Pseudostellariae cyclic peptide extract of Comparative Example 5 and Comparative Example 1 is that the Pseudostellariae cyclic peptide extract of Comparative Example 5 is produced in Nanjing.

[0032] The difference between the preparation method of the Pseudostellariae Radix cyclopeptide extract of Comparative Example 6 and that of Comparative Example 2 is that the Pseudostellariae Radix in Comparative Example 6 is produced in Nanjing.

[0033] The preparation method of the Pseudostellariae cyclic peptide extract of Comparative Example 7 is different from that of Comparative Example 3 in that the Pseudostellariae cyclic peptide extract of Comparative Example 7 is produced in Nanjing.

[0034] The difference between the preparation method of the Pseudostellariae Radix cyclopeptide extract of Comparative Example 8 and that of Comparative Example 4 is that the Pseudostellariae Radix in Comparative Example 8 is produced in Nanjing.

[0035] The extraction rates of the Pseudostellariae cyclic peptide extracts of Examples 1 to 6 and Comparative Examples 1 to 8 are shown in Table 1.

[0036] Table 1 Extraction rate of Pseudostellariae cyclic peptide extracts of Examples 1 to 6 and Comparative Examples 1 to 8

[0037] sample Extraction rate of Pseudostellaria heterophylla cyclic peptide extract (%) Example 1 95.2 Example 2 96.7 Example 3 94.9 Example 4 98.2 Example 5 97.6 Example 6 95.8 Comparative Example 1 90.3 Comparative Example 2 91.1 Comparative Example 3 88.7 Comparative Example 4 76.5 Comparative Example 5 87.7 Comparative Example 6 80.3 Comparative Example 7 86.9 Comparative Example 8 80.2 As can be seen from Table 1, the extraction rate of the Pseudostellaria radix cyclic peptide extract using the composite bacterial agent of Bacillus subtilis and Clostridium butyricum is much higher than that of the Pseudostellaria radix cyclic peptide extract using a single bacteria, and when the mass ratio of Bacillus subtilis and Clostridium butyricum is 1:4, the extraction rate will also decrease. At the same time, after replacing the composite bacterial agent with a composite bacterial agent of Bacillus subtilis and Lactobacillus, the extraction rate of the Pseudostellaria radix cyclic peptide extract is even lower. When Clostridium butyricum is replaced by Lactobacillus, there is no complex effect between Bacillus subtilis and Lactobacillus. Compared with single bacterial fermentation, the extraction rate of the Pseudostellaria radix cyclic peptide extract extracted using Bacillus subtilis and Lactobacillus fermentation is even lower, indicating that there is a complex effect between Bacillus subtilis and Clostridium butyricum.

[0038] The contents of Pseudostellariae cyclic peptide A and Pseudostellariae cyclic peptide B in Pseudostellariae cyclic peptides were detected by Hewlett Packard HP Series 1100 high performance liquid chromatograph. The contents of Pseudostellariae cyclic peptide A and Pseudostellariae cyclic peptide B in Examples 1 to 3 and Comparative Examples 1 to 4 are shown in Table 2, and the contents of Pseudostellariae cyclic peptide A and Pseudostellariae cyclic peptide B in Examples 4 to 6 and Comparative Examples 5 to 8 are shown in Table 3.

[0039] Table 2 Determination results of the contents of Pseudostellariae Cyclic Peptide A and Pseudostellariae Cyclic Peptide B in Examples 1 to 3 and Comparative Examples 1 to 4 sample Pseudostellariae cyclic peptide A (μg / g) Pseudostellariae cyclic peptide B (μg / g) Example 1 206.1 292.7 Example 2 239.2 287.5 Example 3 235.9 296.1 Comparative Example 1 104.3 180.3 Comparative Example 2 187.6 187.9 Comparative Example 3 155.8 163.5 Comparative Example 4 173.7 209.1 As can be seen from Table 2, in the Pseudostellaria zephyranthes cyclic peptide extract extracted by fermentation using the composite bacteria of Bacillus subtilis and Clostridium butyricum of the present invention, the contents of Pseudostellaria zephyranthes cyclic peptide A and Pseudostellaria zephyranthes cyclic peptide B are significantly increased compared with the Pseudostellaria zephyranthes cyclic peptide extract fermented using a single bacteria fermentation. At the same time, compared with the Pseudostellaria zephyranthes cyclic peptide extract extracted by fermentation using the composite bacteria of Bacillus subtilis and Lactobacillus in Comparative Example 4, the contents of Pseudostellaria zephyranthes cyclic peptide A and Pseudostellaria zephyranthes cyclic peptide B in the cyclic peptide extract extracted by fermentation using the composite bacteria of Bacillus subtilis and Clostridium butyricum are also significantly increased, indicating that Bacillus subtilis and Clostridium butyricum in the present invention have a compounding effect.

[0040] Table 3 Determination results of the contents of Pseudostellariae Cyclic Peptide A and Pseudostellariae Cyclic Peptide B in Examples 4 to 6 and Comparative Examples 5 to 8 sample Pseudostellariae cyclic peptide A (μg / g) Pseudostellariae cyclic peptide B (μg / g) Example 4 135.1 98.1 Example 5 129.6 105.8 Example 6 129.4 112.5 Comparative Example 5 67.4 76.3 Comparative Example 6 89.3 82.0 Comparative Example 7 65.2 70.3 Comparative Example 8 79.3 77.4 It can be seen from Tables 2 and 3 that the preparation method of the Pseudostellariae cyclic peptide extract of the present invention is applicable to Pseudostellariae radix from different sources, and can increase the content of Pseudostellariae cyclic peptide A and Pseudostellariae cyclic peptide B in Pseudostellariae radix. The preparation method of the Pseudostellariae cyclic peptide extract of the present invention is applicable to Pseudostellariae radix from different sources.

[0041] Application Example 1 An anti-wrinkle skin care product comprises the following ingredients: 2.5 kg of the Pseudostellariae cyclic peptide extract extracted in Example 1, 1 kg of glycerin, 0.5 kg of allantoin, 3 kg of sodium hyaluronate, 1.5 kg of squalane, 2 kg of isononyl isononanoate, 0.8 kg of cetearyl alcohol, 0.5 kg of polyglyceryl-10 stearate, 1.5 kg of xanthan gum, and 0.01 kg of parahydroxyacetophenone were added and made up to 100 kg with deionized water.

[0042] The preparation method of the anti-wrinkle skin care product of Application Example 1 is as follows: The formulated amounts of glycerin, allantoin, sodium hyaluronate, xanthan gum, and water were stirred uniformly at 50°C and 300 rpm to obtain phase A; the formulated amounts of squalane, isononyl isononanoate, cetearyl alcohol, and polyglyceryl-10 stearate were stirred uniformly at 50°C and 300 rpm to obtain phase B, which was added to phase A, emulsified and homogenized, cooled to 25°C, and the Pseudostellaria heterophylla cyclic peptide extract and p-hydroxyacetophenone of Example 1 were added and stirred uniformly to obtain the product.

[0043] Application Examples 2 to 9 The preparation methods of the skin care products of Application Examples 2 to 9 are roughly the same as those of the skin care products of Application Example 1. The difference between the skin care products of Application Examples 2 to 9 and Application Example 1 is that the Pseudostellaria baicalensis cyclic peptide extracts in Application Examples 2 to 9 are replaced by the Pseudostellaria baicalensis cyclic peptide extracts of Comparative Examples 1 to 8.

[0044] Test Example 1 1. Test substance: the anti-wrinkle skin care products of Application Examples 1 to 9; Blank: a blank group without any substance added.

[0045] 2. Based on: 2015 edition of the "Technical Specifications for Safety of Cosmetics", GB 17149.1-1997 "Diagnostic Criteria and Treatment Principles for Cosmetic Dermatitis", and GB 17149.2-1997 "Diagnostic Criteria and Treatment Principles for Cosmetic Contact Dermatitis".

[0046] 3. Methods: 270 volunteers were divided into 9 groups, 30 in each. The skin on the flexural side of the volunteers' forearms was used as the test site. Test substances and blanks were patch tested for 24 hours according to the method specified in the 2015 edition of the "Safety Technical Specifications for Cosmetics." Skin reactions were observed 30 minutes, 24 hours, and 48 hours after removal of the test substance patch, and the results were recorded.

[0047] 4. Test conclusion: The results of the human skin occlusive patch test showed that the anti-wrinkle skin care products prepared in Examples 1 to 9 had no positive reactions among 270 people, and the test substances did not cause adverse reactions to the skin of this batch of volunteers.

[0048] Test Example 2 After cleansing their faces in the morning and evening, volunteers applied 2 g of the anti-wrinkle skincare product and massaged for 2-3 minutes. All volunteers underwent wrinkle reduction testing on days 0, 30, and 60 after cleansing. The wrinkle reduction test used the VISIA-CR facial image analyzer to capture skin images and measure the wrinkle percentage. The wrinkle percentage difference refers to the difference between the wrinkle percentage after using the anti-wrinkle skincare product of the present invention and the wrinkle percentage before and after use.

[0049] Table 4 Wrinkle removal test results

[0050] As can be seen from Table 4, the wrinkle reduction rate of volunteers using the anti-wrinkle skin care product of Application Example 1 of the present invention is relatively high, indicating that the contents of Pseudostellariae cyclopeptide A and Pseudostellariae cyclopeptide B in the Pseudostellariae cyclopeptide extract after fermentation with the composite bacterial agent of the present invention are increased, thereby achieving a good anti-wrinkle effect.

Claims

1. A method for preparing a Pseudostellariae cyclic peptide extract, characterized in that: The following steps are involved: The Radix Pseudostellariae is crushed to obtain Radix Pseudostellariae powder, the Radix Pseudostellariae powder is soaked in water to obtain Radix Pseudostellariae soaking liquid, a composite bacterial agent is inoculated into the Radix Pseudostellariae soaking liquid for fermentation and culture to obtain a fermentation product, and then the Radix Pseudostellariae cyclic peptide extract is extracted to obtain the Radix Pseudostellariae cyclic peptide extract after freeze-drying; the composite bacterial agent is a composite bacterial agent of Bacillus subtilis and Clostridium butyricum, and the mass ratio of Bacillus subtilis to Clostridium butyricum in the composite bacterial agent is 1:1 to 3.

2. The method for preparing the Pseudostellariae cyclic peptide extract according to claim 1, wherein The inoculation amount of the composite bacterial agent is 10-15%.

3. The method for preparing the Pseudostellariae cyclic peptide extract according to claim 1, wherein: The fermentation temperature is 33-37° C., the fermentation time is 30-50 h, and the pH of the fermentation is 6-8.

4. The method for preparing the Pseudostellariae cyclic peptide extract according to claim 1, wherein: The extraction method comprises the following steps: extracting on a constant temperature shaker at 30-40° C. and a rotation speed of 200-300 rpm for 0.5-1 h, followed by centrifugation at 5000-6000 r / min for 10-30 min.

5. The method for preparing the Pseudostellariae cyclic peptide extract according to claim 1 or 4, characterized in that: Before extraction, the concentration of the fermentation product is diluted to 30-50 g / L.

6. The method for preparing the Pseudostellariae cyclic peptide extract according to claim 1, wherein: The mass volume ratio of the Pseudostellariae Radix powder to water is 1 g: 10-15 mL, and the soaking time is 30-90 min.

7. The method for preparing the Pseudostellariae cyclic peptide extract according to claim 1, wherein: The mesh size of the Pseudostellaria heterophylla powder is 20 to 30 meshes.

8. The method for preparing the Pseudostellariae cyclic peptide extract according to claim 1, wherein: The freeze-drying method comprises: pre-freezing at normal pressure and -30 to -50°C for 30 to 60 minutes, and then drying at -70 to -80°C in vacuum for 30 to 60 minutes.

9. A Pseudostellariae cyclic peptide extract, characterized in that: The extract is prepared by the preparation method of the Pseudostellaria baicalensis cyclic peptide extract according to claim 1.

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