Soothing agent containing piper nigrum extract as well as preparation method and application of soothing agent

Abstract

The invention belongs to the field of cosmetics and the field of plant extracts, and provides a soothing agent containing a piper nigrum extract and a preparation method and application thereof, the soothing agent contains the piper nigrum extract, and the preparation method of the piper nigrum extract comprises the following steps: (1) raw material treatment; (2) performing enzymolysis treatment; and (3) ethanol extraction. The Kava pepper extract is prepared by using an enzymolysis method, and the obtained soothing agent is high in content of effective components, has good safety, soothing activity and antioxidant activity, and can be used for various cosmetics.

Description

Technical Field

[0001] This application pertains to the fields of cosmetics and plant extracts. Specifically, this application provides a soothing agent containing kava pepper extract, its preparation method, and its application. Background Technology

[0002] Kava pepper ( Piper methysticum Also known as Kava, its main components are kava lactone compounds (which also include alkaloids, amides and other active ingredients), which have sedative, anti-anxiety, hypnotic, muscle relaxant, antimicrobial, anti-inflammatory and skin-soothing effects.

[0003] Kava extract has become one of the world's best-selling herbal medicines, widely used in pharmaceuticals, health products, and cosmetics. However, for many years, the preparation method of kava extract has mainly used the common alcohol extraction method, without significant progress in extraction efficiency and product efficacy. Summary of the Invention

[0004] On the one hand, this application provides a soothing agent containing kava pepper extract, wherein the soothing agent contains kava pepper extract and the preparation method of kava pepper extract includes: (1) raw material treatment; (2) enzymatic hydrolysis treatment; and (3) ethanol extraction.

[0005] Further, the raw material processing in step (1) is as follows: washing, drying and pulverizing the roots, stems and / or leaves of kava pepper into powder.

[0006] Further, the enzymatic hydrolysis in step (2) is as follows: add 4-6 times the weight of water, 600-800 U / mL cellulase, 600-800 U / mL hemicellulase, 600-800 U / mL pectinase, 400-600 U / mL inulinase, and 0.4-0.6 w / v% pentylrhamnoside to the powder obtained in step (1); adjust the pH to 7-8, and enzymatically hydrolyze at 35-45 degrees Celsius for 1-2 hours; filter and collect the precipitate.

[0007] Further, the enzymatic hydrolysis in step (2) is as follows: add 5 times the weight of water, 700 U / mL cellulase, 700 U / mL hemicellulase, 700 U / mL pectinase, 500 U / mL inulinase, and 0.5 w / v% pentylrhamnoside to the powder obtained in step (1); adjust the pH to 7-8, and enzymatically hydrolyze at 40°C for 1-1.5 hours; filter and collect the precipitate.

[0008] Further, the ethanol extraction in step (3) is as follows: 15-30 times the mass of 80-85 v / v% ethanol of the powder obtained in step (1) is added to the precipitate obtained in step (2); the extract is extracted at 60-70°C for 3-4 hours to obtain an extract; the extract is filtered and concentrated under reduced pressure to obtain an extract paste; the extract paste is thoroughly dried to obtain the kava pepper extract.

[0009] Further, the ethanol extraction in step (3) is as follows: 20 times the mass of 80 v / v% ethanol of the powder obtained in step (1) is added to the precipitate obtained in step (2); the extract is extracted at 70°C for 3 hours to obtain an extract; the extract is filtered and concentrated under reduced pressure at 40°C and a vacuum degree of less than 0.08 MPa to obtain an extract paste; the extract paste is thoroughly dried to obtain the kava pepper extract.

[0010] Furthermore, the soothing agent also includes caprylic / capric triglycerides.

[0011] Furthermore, the ratio of kava pepper extract and caprylic / capric triglycerides in the soothing agent is calculated as 1:6-10 in g:mL.

[0012] Depending on the type of cosmetic, the expected amount added, and the required effect, those skilled in the art can routinely adjust the proportions of the various ingredients in the soothing agent, preferably with a capsaicin content greater than 1500 ppm (0.15 w / w%).

[0013] The soothing agent of this application can be compounded with various compatible cosmetic ingredients, preferably for use in cosmetics with anti-allergic, soothing, and antioxidant functions, and preferably mixed with other ingredients at a temperature below 45°C.

[0014] On the other hand, this application provides the application of the above-mentioned soothing agent in cosmetics, wherein the cosmetics contain 0.05-5 w / w% of the above-mentioned soothing agent.

[0015] Furthermore, the cosmetic contains 0.1-2 w / w% of the aforementioned soothing agent.

[0016] On the other hand, this application provides a method for preparing the above-mentioned soothing agent, the method including the preparation method of kava pepper extract in the above technical solution.

[0017] Furthermore, the method also includes the step of mixing kava pepper extract with caprylic / capric triglyceride.

[0018] By designing a reasonable enzymatic hydrolysis system, especially by adding surfactants, this application effectively improved the extraction efficiency and active ingredient content of kava leaves / roots / stems, and prepared a soothing product with good safety, soothing activity and antioxidant activity. Attached Figure Description

[0019] Figure 1 The extraction rates at different temperatures in Example 2 are shown.

[0020] Figure 2 The content of various calvalactone compounds in the extracts at different temperatures in Example 2 is shown.

[0021] Figure 3 The extraction rates are for different enzymatic hydrolysis schemes in Example 3.

[0022] Figure 4 The content of various calvaline compounds in the extracts of different enzymatic hydrolysis schemes in Example 3 is shown.

[0023] Figure 5 The value represents the PGE-2 content (0.2%) in RAW264.7 cells induced by LPS.

[0024] Figure 6 NO content (0.2%) in RAW264.7 cells induced by LPS.

[0025] Figure 7 The value represents the IL-6 content (0.2%) in RAW264.7 cells induced by LPS.

[0026] Figure 8 The value represents the TNF-α content (0.2%) in RAW264.7 cells induced by LPS.

[0027] Figure 9 The results are from the DPPH clearance rate experiment.

[0028] Figure 10 The results of the chicken embryo chorioallantoic membrane experiment (6%). Detailed Implementation Example 1: Basic Extraction and Detection Methods

[0029] The kava pepper roots (from Shaanxi Ruichang Biotechnology Co., Ltd.) were washed, dried, and pulverized into kava pepper root powder. Add 80 v / v% ethanol to the Kava pepper root powder at a material-to-liquid ratio of 1:20 (g: mL) and extract at a predetermined temperature for 3 hours to obtain the extract. The extract was filtered and concentrated under reduced pressure at 40°C and a vacuum degree of less than 0.08 MPa to obtain an extract paste. The extract was thoroughly dried and pulverized.

[0030] The contents of capsanthin, dihydrocapsanthin, capsanthin, dihydrocapsanthin, demethoxycapsanthin, and methoxycapsanthin in the extract were determined using an Agilent 1100 HPLC system (equipped with an HC-C18 column, column temperature: 25℃, flow rate: 0.8mL / min, wavelength: 256nm) and standards provided by Shanghai Yuanye. Example 2: Effect of different extraction temperatures on extraction efficiency

[0031] like Figure 1 As shown, the extraction rate is calculated based on the ratio of extract mass to raw material mass. The extraction rate increases slightly at higher temperatures, especially between 60-80 degrees Celsius.

[0032] The results are as follows Figure 2 As shown, the percentage content of capsaicin and dihydrocapsaicin did not differ much at various temperatures, while the percentage content of anesthetic capsaicin decreased significantly (from 7-8% to 3-4%). ​​The proportion of calvalactone compounds, which are of concern, in the total extract also decreased from nearly 70% to 55%.

[0033] A similar trend is observed when extracting from stems and leaves with lower calvaline content. That is, during prolonged extraction, there is a certain contradiction between the instability of some calvaline compounds at high temperatures and the need to increase the total yield. Example 3: Effect of adding an enzyme treatment step

[0034] The applicant attempted to add an enzymatic hydrolysis step before the ethanol extraction step. Five times the mass of water and enzyme were added to the kava pepper root powder, the pH was adjusted to 7.5 for which most enzymes have good activity, and enzymatic hydrolysis was carried out at 40 degrees Celsius for 1.5 hours (to avoid the high temperature affecting the kava lactone compounds and the water carrying away too many components, a higher amount of enzyme and a lower amount of water, temperature and time were selected; the subsequent high concentration of ethanol can automatically inactivate the enzyme), the precipitate was filtered and extracted according to the method of Example 1 (70 degrees Celsius).

[0035] The enzymatic hydrolysis system used includes: Enzymatic hydrolysis system 1: cellulase (700 U / mL), hemicellulase (700 U / mL), pectinase (700 U / mL).

[0036] Enzymatic hydrolysis system 2: cellulase (700U / mL), hemicellulase (700U / mL), pectinase (700U / mL), fig protease (500U / mL), bromelain (500U / mL).

[0037] Enzymatic hydrolysis system 3: cellulase (700 U / mL), hemicellulase (700 U / mL), pectinase (700 U / mL), xylanase (700 U / mL).

[0038] Enzymatic hydrolysis system 4: cellulase (700 U / mL), hemicellulase (700 U / mL), pectinase (700 U / mL), inulinase (500 U / mL).

[0039] Enzymatic hydrolysis system 5: cellulase (500 U / mL), hemicellulase (500 U / mL), pectinase (500 U / mL), α-galactosidase (500 U / mL).

[0040] Extraction rate, such as Figure 3 As shown, although the enzymatic hydrolysis system removes some water-soluble components, the extraction rate after enzymatic hydrolysis of systems 3 and 4 still exceeds 11% (which is basically the same as or even slightly higher than that at 70°C in Example 2), meaning that enzymatic hydrolysis effectively helps release the contents of the raw materials.

[0041] The content of various calvalactones in the extract, such as Figure 4 As shown, with Figure 2 Compared with the 60℃ group, the proportion of calvalactone compounds in the total extract increased from about 56% to about 60%, which is mainly attributed to the increase of naringin compounds (from 3-4% to about 6%). Example 4: Adding a surfactant to the enzymatic hydrolysis system

[0042] Considering that the main focus is on non-polar or low-polar components in kava pepper, the applicant further added 0.5 w / v% of polysorbate-20, pentyl rhamnoside, and cocoyl betaine to the enzymatic hydrolysis systems 3 and 4 of Example 3.

[0043] The results are shown in Tables 1 and 2 below: Table 1. Extraction rate and calvanolide content of the enzymatic hydrolysis system after adding surfactant.

[0044] Table 2. Content of various calvanolides in the 4+pentylrhamnoside enzymatic hydrolysis system.

[0045] The results showed that the enzymatic hydrolysis system using 4+pentylrhamnoside effectively improved the extraction rate (to nearly 13%) and calvanolide content (over 70%), with increases in the content of various calvanolide compounds, particularly the previously significantly reduced naringin compounds, achieving a doubling effect compared to direct extraction. This may be due to the addition of surfactants altering the way calvanolide-containing structures exist in the raw materials, thereby increasing the release or protection of calvanolide.

[0046] When this method is applied to the roots and stems of kava pepper, the product extraction rate can reach over 12%, and kava lactone accounts for more than 63% of the extract.

[0047] Taking all factors into account, the extraction efficiency of kavalactone is about 1.7 times that of the original alcohol extraction method, and it can achieve high purity without the need for costly and slow chromatography methods. Example 5: Practical effects of a soothing agent containing kava pepper extract

[0048] The extract can be formulated into a soothing product with ingredients such as caprylic / capric triglycerides. For example, the ratio of kava extract to caprylic / capric triglycerides in the soothing product is 1:6-10, preferably 1:8, calculated in g:mL.

[0049] The applicant commissioned a testing agency (Testing Center of the School of Biomedical Sciences, Guangdong University of Technology) to test the product's soothing effect.

[0050] The effects of LPS-induced PGE-2, NO, IL-6, and TNF-α levels on RAW264.7 cells, as well as the DPPH free radical scavenging activity and stimulatory effect, were investigated. The results are as follows: Figure 5-10 As shown.

[0051] The results show: The concentration of the sample kava pepper extract was determined to be 0.2% by cytotoxicity assay, with a corresponding cell survival rate of 99.94% ± 1.09%.

[0052] Compared with the NC group, the PGE-2 content of the kava pepper extract was significantly decreased (p<0.0001), with a PGE-2 inhibition rate of 35.38%±6.06%, indicating that the kava pepper extract inhibited the secretion of PGE-2 inflammatory factors. Compared with the NC group, the NO content of the kava pepper extract was significantly decreased (p<0.0001), with a NO inhibition rate of 74.84%±0.11%, indicating that the kava pepper extract inhibited the secretion of NO inflammatory factors. Compared with the NC group, the IL-6 content of the kava pepper extract was significantly decreased (p<0.05), with an IL-6 inhibition rate of 32.84%±0.91%, indicating that the kava pepper extract inhibited the secretion of IL-6 inflammatory factors. Compared with the NC group, the TNF-α content of the kava pepper extract was significantly decreased (p<0.01), with a TNF-α inhibition rate of 25.56%±10.45%, indicating that the kava pepper extract inhibited the secretion of TNF-α inflammatory factors. At concentrations of 10% and 20% v / v, the DPPH free radical scavenging rate reached approximately 24% and 30%, respectively. Chicken embryo chorioallantoic membrane experiments showed that even concentrations as high as 6% did not produce a significant irritant effect.

[0053] The results showed that the product is safe and has good anti-inflammatory, soothing and antioxidant effects.

Claims

1. A soothing agent containing kava pepper extract, characterized in that, The soothing agent contains kava pepper extract, and the preparation method of the kava pepper extract includes: (1) raw material treatment; (2) enzymatic hydrolysis; and (3) ethanol extraction.

2. The soothing agent according to claim 1, wherein the raw material processing in step (1) is as follows: washing, drying and pulverizing the roots, stems and / or leaves of kava pepper into powder.

3. The soothing agent according to claim 1 or 2, wherein the enzymatic hydrolysis in step (2) is as follows: add 4-6 times the weight of the powder of water, 600-800 U / mL of cellulase, 600-800 U / mL of hemicellulase, 600-800 U / mL of pectinase, 400-600 U / mL of inulinase, and 0.4-0.6 w / v% of pentylrhamnoside to the powder obtained in step (1); adjust the pH to 7-8, and enzymatically hydrolyze at 35-45 degrees Celsius for 1-2 hours; filter and collect the precipitate.

4. The soothing agent according to claim 3, wherein the enzymatic hydrolysis in step (2) is as follows: add 5 times the weight of water, 700 U / mL cellulase, 700 U / mL hemicellulase, 700 U / mL pectinase, 500 U / mL inulinase, and 0.5 w / v% pentylrhamnoside to the powder obtained in step (1); adjust the pH to 7-8, and enzymatically hydrolyze at 40°C for 1-1.5 hours; filter and collect the precipitate.

5. The soothing agent according to any one of claims 1-4, wherein the ethanol extraction (3) is performed by adding 15-30 times the mass of 80-85 v / v% ethanol of the powder obtained in step (1) to the precipitate obtained in step (2); extracting at 60-70°C for 3-4 hours to obtain an extract; filtering the extract and concentrating under reduced pressure to obtain an extract paste; and drying the extract paste thoroughly to obtain the kava pepper extract.

6. The soothing agent according to claim 5, wherein the ethanol extraction (3) is performed by adding 20 times the mass of 80 v / v% ethanol of the powder obtained in step (1) to the precipitate obtained in step (2); extracting at 70°C for 3 hours to obtain an extract; filtering the extract and concentrating it under reduced pressure at 40°C and a vacuum degree of less than 0.08 MPa to obtain an extract paste; and drying the extract paste thoroughly to obtain the kava pepper extract.

7. The soothing agent according to claim 1, wherein the soothing agent further comprises caprylic / capric triglyceride.

8. The soothing agent according to claim 7, wherein the ratio of kava pepper extract and caprylic / capric triglyceride in the soothing agent is calculated as 1:6-10 in g:mL.

9. The application of the soothing agent according to any one of claims 1-8 in a cosmetic, wherein the cosmetic contains 0.05-5 w / w% of the soothing agent according to any one of claims 1-8.

10. The application according to claim 9, wherein the cosmetic contains 0.1-2 w / w% of the soothing agent according to any one of claims 1-8.

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