Gentiana macrophylla extract as well as extraction process, application and skin care product thereof

Abstract

The invention discloses a gentiana macrophylla extract, an extraction process and application thereof and a skin care product. The invention belongs to the field of new daily chemical materials, and discloses an extraction process of a gentiana macrophylla extract, which comprises the following steps: step 1, crushing gentiana macrophylla, dispersing the crushed gentiana macrophylla in water, and carrying out enzymolysis by using cellulase to obtain an enzymolysis product; 2, filtering the enzymolysis product, collecting filtrate and filter residues, and drying the filter residues to obtain micro powder; performing vacuum concentration and alcohol precipitation on the filtrate, and collecting to obtain a first extract; step 3, carrying out alcohol extraction on the micro powder once or multiple times, combining extracting solutions, and concentrating the extracting solutions to obtain thick paste; 4, drying the thick paste to prepare powder, so as to obtain a second extract; and step 5, combining the first extract and the second extract to obtain the gentiana macrophylla extract. Through detection, the ratio of loganic acid to gentiopicroside is 1: 3.94, and the compound contains a small amount of swertiamarin, and after related inflammatory factors and other properties are detected, the performance of the compound is better. Meanwhile, the invention also provides related application of the extract.

Description

Technical Field

[0001] The present invention relates to the field of new daily chemical materials, and in particular to a gentiana macrophylla extract, an extraction process thereof, uses thereof, and a skin care product. Background Art

[0002] Gentiana macrophylla extract has been widely used in cosmetics to soothe skin. Its main functional ingredients are gentiopicroside, loganinic acid, swetimarin, and swetirin. Gentipicroside's main effects include reducing inflammation, providing anti-inflammatory and soothing effects, providing antioxidant and repair benefits, while loganinic acid's main effects include triple antibacterial (anti-inflammatory, anti-oxidant), moisturizing and soothing effects.

[0003] The patent application with publication number CN112274461A, which is titled "Gentiana macrophylla extract, preparation method and use thereof", discloses a Gentiana macrophylla extract prepared by an alcohol extraction method;

[0004] In this patent application, the concentrations of loganinic acid and gentiopicroside obtained by water extraction, 60 vol% alcohol extraction, and 75 vol% alcohol extraction were compared. Among them, the extraction efficiency of loganinic acid and gentiopicroside was improved by alcohol extraction compared with water extraction; the ratio of loganinic acid to gentiopicroside was approximately 2:7.

[0005] The main purpose and research direction of this case is to further study Gentiana macrophylla and further expand its application scope. Summary of the Invention

[0006] One of the purposes of the present invention is to provide an extraction process for a gentiana macrophylla extract, in which gentiana macrophylla powder is pretreated by enzymatic hydrolysis and then subjected to alcohol extraction to obtain the gentiana macrophylla extract. After testing, the ratio of loganin acid to gentiopicroside is 1:3.94, and the extract contains a small amount of scutellariamarin. When the extract is tested for related inflammatory factors and other properties, it is found that the extract has better performance.

[0007] At the same time, the invention also provides related applications of the extract.

[0008] To achieve the above object, the present invention provides an extraction process for a Gentiana macrophylla extract, comprising the following steps:

[0009] Step 1: crushing the Gentiana macrophylla and dispersing it in water, and performing enzymolysis with cellulase to obtain an enzymolysis product;

[0010] Step 2: Filter the enzymatic hydrolysis product to collect the filtrate and filter residue, and dry the filter residue to obtain a micropowder; after the filtrate is concentrated under reduced pressure and precipitated with alcohol, the first extract is collected;

[0011] Step 3: extract the micropowder with alcohol once or multiple times, combine the extracts, and concentrate the extracts to obtain a thick paste;

[0012] Step 4: Drying and powdering the thick paste to obtain a second extract;

[0013] Step 5: Combine the first extract and the second extract to obtain the Gentiana macrophylla extract.

[0014] In the above extraction process of Gentiana macrophylla extract, in step 3, 95 vol% ethanol is used for the alcohol extraction, and the weight-to-volume ratio of the micropowder to ethanol is 1:10 g / ml.

[0015] In the above extraction process of Gentiana macrophylla extract, the number of alcohol extraction operations is 2 to 3 times, the single alcohol extraction time is 1 to 2 hours, and the alcohol extraction temperature is 50 to 65°C.

[0016] In the above-mentioned extraction process of Gentiana macrophylla extract, in the step 1, the Gentiana macrophylla is crushed and passed through a 300-mesh sieve, and then mixed with water in a mass-to-volume ratio of 1:5-10, and 0.1-0.2 wt% of cellulase equivalent to the weight of Gentiana macrophylla is added for enzymatic hydrolysis;

[0017] The enzymatic hydrolysis time is 2 to 3 hours, and the enzymatic hydrolysis temperature is 45 to 65°C.

[0018] At the same time, the present invention also provides a Gentiana macrophylla extract, which is prepared by any of the above methods.

[0019] In addition, the present invention also discloses the use of the gentiana macrophylla extract in preparing cosmetics; the cosmetics are cosmetics that can soothe and / or alleviate allergies after acting on the skin.

[0020] Finally, the present invention also discloses a cosmetic containing the above-mentioned Gentiana macrophylla extract, with a recommended dosage of 1 to 2 wt%.

[0021] Beneficial effects

[0022] The present invention pre-treats gentiana micropowder by enzymatic hydrolysis and then performs alcohol extraction to obtain the gentiana extract; after testing, the ratio of loganin acid to gentiopicroside is 1:3.94, and when relevant inflammatory factors and other properties are tested, it is found that the performance is better. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1a This is the detection spectrum of Gentiana macrophylla extract 1;

[0024] Figure 1b This is the detection spectrum of Gentiana macrophylla extract 2;

[0025] Figure 2 This is the cytotoxicity test result of Gentiana macrophylla extract 1;

[0026] Figure 3This is the capsaicin receptor (TRPV1) test result of Gentiana macrophylla extract 1;

[0027] Figure 4 These are the results of volunteer tests. DETAILED DESCRIPTION

[0028] The present invention will be further described below in conjunction with the embodiments, but this does not constitute any limitation to the present invention. Any limited modifications made within the scope of the claims of the present invention are still within the scope of the claims of the present invention.

[0029] Example 1

[0030] An extraction process for a Gentiana macrophylla extract comprises the following steps:

[0031] Step 1: Grind the gentiana macrophylla and pass it through a 300-mesh sieve, then mix it with water at a mass-to-volume ratio of 1:10, add 0.15 wt% of cellulase (β-1,4-glucan-4-glucanohydrolase, 100,000 u / g) equivalent to the weight of the gentiana macrophylla for enzymatic hydrolysis, the enzymatic hydrolysis time is 3 h, the enzymatic hydrolysis temperature is 60°C, and obtain an enzymatic hydrolysis product;

[0032] Step 2: Filter the enzymatic hydrolysis product to collect the filtrate and filter residue, and dry the filter residue to obtain a micropowder; after the filtrate is concentrated under reduced pressure and precipitated with alcohol, the first extract is collected;

[0033] More specifically, the filtrate is concentrated under reduced pressure to obtain a thick paste, which is added to 10 times the volume of a 90 vol% ethanol solution for alcohol precipitation and filtration, and the filtrate is collected and concentrated under reduced pressure, and then powdered and dried to obtain the first extract;

[0034] Step 3: Extract the micropowder with alcohol three times, combine the extracts, and concentrate the extracts to obtain a thick paste;

[0035] The single alcohol extraction time was 100 min, the weight-to-volume ratio of micropowder to ethanol was 1:10 g / ml, and the alcohol extraction temperature was 65°C.

[0036] Step 4: Drying and powdering the thick paste obtained in step 3 to obtain a second extract;

[0037] Step 5: Combine the first extract and the second extract to obtain Gentiana macrophylla extract 1.

[0038] Comparative Example 1

[0039] An extraction process for a Gentiana macrophylla extract comprises the following steps:

[0040] Step 1: Grind the Gentiana macrophylla and pass it through a 300-mesh sieve to obtain fine powder;

[0041] Step 2: Extract the micropowder with alcohol three times, combine the extracts, and concentrate the extracts to obtain a thick paste;

[0042] The single alcohol extraction time was 100 min, the weight-to-volume ratio of micropowder to ethanol was 1:10 g / ml, and the alcohol extraction temperature was 65°C.

[0043] Step 3: Dry and powder the thick paste obtained in step 2 to obtain Gentiana macrophylla extract 2.

[0044] Ingredient testing

[0045] Gentiana macrophylla extract 1 and Gentiana macrophylla extract 2 were sent to Guangxi Zhongle Health Industry Co., Ltd. for liquid chromatography detection.

[0046] Detection spectrum see Figure 1a and Figure 1b ;

[0047] The test results are shown in Table 1 below;

[0048] Table 1 Composition of Gentiana macrophylla extract Content: wt%

[0049] Loganinic acid Gentiopicroside Swertiamarin Gentiana macrophylla extract 1 13.79 54.38 6.44 Gentiana macrophylla extract 2 40.2 25.03 0.17

[0050] Figure 1a The peak position of loganinic acid is 13.3min, the peak position of swertiamarin is 22.07min, and the peak position of gentiopicroside is 32.5min. Figure 1b Among them, the peak position of loganinic acid is 12.49 min, the peak position of swedemarin is 22.23 min, and the peak position of gentiopicroside is 30.8 min.

[0051] Security Testing

[0052] The solution containing Gentiana macrophylla extract 1 (G-repair Xinyuening, containing Gentiana macrophylla extract 1: 1 wt%) was sent to Shaanxi Boxi General Testing Technology Co., Ltd. for testing. The test report number is: G3021-2503110;

[0053] Detection method: Cytotoxicity assay based on U266 cells;

[0054] 1) Cell seeding: After thawing cells, when the plating rate reaches about 60%, seed the cells into a 96-well plate and incubate in a CO2 incubator (37°C, 5% CO2) overnight.

[0055] 2) Experimental Grouping: The experiment was divided into a zero adjustment group, a solvent control group, a positive control group, and a sample group. In the sample group, each sample was set up with 8 concentration gradients, and 3 replicate wells were set up under each concentration gradient.

[0056] 3) Solution preparation: Prepare sample working solutions of different concentrations according to the test concentration setting table.

[0057] According to the results of CCK8 assay, G-repair Xinyuening showed no significant cytotoxicity in U266 cells within the concentration range of 0.3125% (v / v); Figure 2 .

[0058] In vitro soothing efficacy test

[0059] The solution containing Gentiana macrophylla extract 1 (G-repair Xinyuening, containing Gentiana macrophylla extract 1: 1 wt%) was sent to Shaanxi Boxi General Testing Technology Co., Ltd. for testing. The test report number is: G3021-2503189;

[0060] Detection method:

[0061] Based on 3D epidermal skin model Detection of capsaicin receptor (TRPV1) and inflammatory factors (TNF-a, IL-1a); Detection of immunoglobulin E (IgE) content based on U266 cells;

[0062] Based on 3D epidermal skin model Test

[0063] The test groups can be seen in Table 2;

[0064] Table 2

[0065]

[0066] Stimulation and drug administration

[0067] 1) According to the test group, transfer the model to a 6-well plate (add 0.9 mL of EpiGrowth culture medium in advance) and mark the test group number on the 6-well plate.

[0068] 2) 25 μL of 0.1% SLS solution was added to the surface of the NC group, PC1 group, PC2 group, and sample group and incubated for 30 min.

[0069] 3) After incubation, add the working solution of corresponding concentration to the PC1 and PC2 groups, and take the working solution of the sample to be tested to the sample group

[0070] ELISA test

[0071] After incubation, collect the 3D epidermal skin model culture medium into a centrifuge tube. Store the sample for ELISA testing at -80°C and perform analysis according to the ELISA kit's instructions. Apply the sample evenly to the model surface and incubate in a CO2 incubator (37°C, 5% CO2) for 24 hours.

[0072] After the incubation, the residual test substance was washed with sterile PBS solution and the residual liquid was wiped away with a sterile cotton swab.

[0073] Immunofluorescence testing

[0074] The model to be tested was fixed with 4% paraformaldehyde for 24 hours, and then immunofluorescence detection was performed. The images were observed under a microscope, and the pictures were collected and analyzed.

[0075] Inhibition rate calculation: Inhibition rate = (negative control group - sample group) * 100% / negative control group

[0076] Capsaicin receptor (TRPV1) test results are available Figure 3 ;

[0077] The above pictures were taken with a fluorescence microscope (Olympus, BX43) at a magnification of 20x. Blue fluorescence represents the cell nucleus, and green fluorescence represents TRPV1. The weaker the green fluorescence intensity, the lower the TRPV1 content.

[0078] The test results can be seen in Table 3;

[0079] Table 3 Test results

[0080]

[0081] Compared with the BC group, the TRPV1 content in the NC group increased significantly, indicating that the stimulation conditions of this test were effective.

[0082] Compared with the NC group, the TRPV1 content in the PC1 group decreased significantly, indicating that the positive control in this test was effective.

[0083] Compared with the NC group, the TRPV1 content of the sample G-repair Xinyuening at a concentration of 1% was significantly decreased, with an inhibition rate of 44.98%.

[0084] The test results of inflammatory factor (TNF-a) refer to Table 4;

[0085] Table 4

[0086]

[0087] Compared with the BC group, the TNF-a content in the NC group increased significantly, indicating that the stimulation conditions of this test were effective.

[0088] Compared with the NC group, the TNF-a content in the PC2 group was significantly decreased, indicating that the positive control in this test was effective.

[0089] Compared with the NC group, the TNF-a content of the sample G-repair Xinyuening at a concentration of 1% was significantly decreased, with an inhibition rate of 44.25%.

[0090] The test results of inflammatory factor (IL-1a) refer to Table 5;

[0091] Table 5

[0092]

[0093] Compared with the BC group, the IL-1a content in the NC group increased significantly, indicating that the stimulation conditions of this test were effective.

[0094] Compared with the NC group, the I-1a content in the PC2 group decreased significantly, indicating that the positive control in this test was effective.

[0095] Compared with the NC group, the Ⅱ-1a content of the sample G-repair Xinyuening at a concentration of 1% was significantly decreased, with an inhibition rate of 29.52%.

[0096] U266 cell-based assay

[0097] Test Method

[0098] Cell seeding

[0099] After thawing the cells, when the plating rate reached about 60%, the cells were seeded into 6-well plates and incubated overnight in a CO2 incubator (37°C, 5% CO2).

[0100] Liquid preparation

[0101] Prepare the test substance working solution according to the test group;

[0102] Please refer to Table 6 for details;

[0103] Table 6

[0104]

[0105] Drug administration

[0106] According to the test grouping, when the cell plating rate in the 6-well plate reaches 30% to 40%, the group administration is carried out, and 3 replicates are set up for each group. 2 ml of culture medium is added to each well of the blank control group, 2 ml of culture medium containing LPS is added to each well of the negative control group, and 2 ml of culture medium containing LPS and dexamethasone is added to each well of the positive control group. 2 ml of culture medium containing LPS and the corresponding concentration of the test sample is added to each well of the sample group. After the administration is completed, the 6-well plate is placed in a CO2 incubator (37°C, 5% CO2) and cultured for 48 hours.

[0107] ELISA test

[0108] After the incubation, the cell culture supernatant was collected into a centrifuge tube and stored in a -80°C refrigerator. The assay was performed according to the instructions of the ELISA assay kit.

[0109] Inhibition rate calculation: Inhibition rate = (negative control group - sample group) * 100% / negative control group

[0110] The test results can be seen in Table 7

[0111] Table 7

[0112]

[0113] Compared with the BC group, the IgE content in the NC group increased significantly, indicating that the stimulation conditions of this test were effective.

[0114] Compared with the NC group, the IgE content in the PC group decreased significantly, indicating that the positive control in this test was effective.

[0115] Compared with the NC group, the IgE content of the sample G-repair Xinyuening at a concentration of 0.3125% was significantly decreased, with an inhibition rate of 52.78%.

[0116] in conclusion

[0117] Based on the 3D epidermal skin model (EpiKutis), compared with the control group, the sample G-repair Xinyuening had significantly decreased capsaicin receptor (TRPV1) and inflammatory factor (TNF-0, IL-1a) contents at a concentration of 1% (vv), with inhibition rates of 44.98%, 44.25% and 29.52%, respectively. This indicates that the sample can reduce the capsaicin receptor (TRPV1) content at this concentration, inhibit the secretion of inflammatory factors (TNF-a, IL-1a), and has a soothing effect.

[0118] Based on U266 cells, compared with the control group, the immunoglobulin E (IgE) content of the sample G-repair Xinyuening at a concentration of 0.3125% (v / v) was significantly decreased, with an inhibition rate of 52.78%, indicating that the sample can reduce the immunoglobulin E (IgE) content at this concentration and has a soothing effect.

[0119] In vitro efficacy testing

[0120] The solution containing Gentiana macrophylla extract 1 (G-repair Xinyuening, containing Gentiana macrophylla extract 1: 1 wt%) was sent to Shaanxi Boxi General Testing Technology Co., Ltd. for testing. The test report number is: G3021-2503180;

[0121] Testing method: Based on 3D epidermal skin model Detection of inflammatory factor (IL-6) content;

[0122] The test groups can be seen in Table 8;

[0123] Table 8

[0124]

[0125] Stimulation and drug administration

[0126] 1) According to the test group, transfer the model to a 6-well plate (add 0.9 mL of EpiGrowth culture medium in advance) and mark the test group number on the 6-well plate.

[0127] 2) 25 μl of 0.1% SLS solution was added to the surface of the NC group, PC group, and sample group and incubated for 30 min.

[0128] 3) After the incubation, add the corresponding concentration of working solution to the PC group solution. For the sample group, add the working solution of the sample to be tested to the model surface, spread it evenly, and then incubate it in a CO2 incubator (37°C, 5% CO2) for 24 hours.

[0129] 4) After the incubation, wash the remaining test substance with sterile PBS solution and wipe away the residual liquid with a sterile cotton swab.

[0130] ELISA test

[0131] After the incubation, the culture medium of the 3D epidermal skin model was collected in a centrifuge tube. After collection, the samples for ELISA detection were placed in a -80°C refrigerator for freezing and stored, and the detection and analysis were performed according to the operating instructions of the ELISA kit.

[0132] Inhibition rate calculation: Inhibition rate = (negative control group - sample group) * 100% / negative control group

[0133] The test results can be seen in Table 9

[0134] Table 9

[0135]

[0136] Compared with the BC group, the I-6 content in the NC group increased significantly, indicating that the stimulation conditions of this test were effective.

[0137] Compared with the NC group, the I-6 content in the PC group decreased significantly, indicating that the positive control in this test was effective.

[0138] Compared with the NC group, the II-6 content of the sample G-repair Xinyuening at a concentration of 1% was significantly decreased, with an inhibition rate of 20.29%.

[0139] in conclusion:

[0140] Based on the 3D epidermal skin model (EpiKutis), compared with the control group, the content of inflammatory factors (IL-6) in the sample G-repair Xinyuening at a concentration of 1% (v / v) was significantly decreased, with an inhibition rate of 20.29%, indicating that the sample can inhibit the secretion of inflammatory factors (IL-6) at this concentration and has a soothing effect.

[0141] Transepidermal water loss (TEWL) value, skin hemoglobin index, and body surface temperature test

[0142] The solution containing Gentiana macrophylla extract 1 (G-repair Xinyuening, containing Gentiana macrophylla extract 1: 1 wt%) was sent to Shaanxi Boxi General Testing Technology Co., Ltd. for testing. The test report number is: G3209-2504015;

[0143] Purpose of the trial

[0144] Adopting the before-and-after control principle and the parallel control principle, 30 people used the samples continuously, and the transepidermal water loss (TEWL) values before using the samples and 30 minutes after using the samples were calculated.

[0145] Adopting the before-and-after control principle and the parallel control principle, 30 people used the samples continuously, and the skin hemoglobin index of the initial value before using the samples and 30 minutes after using the samples was calculated.

[0146] Adopting the before-and-after control principle and the parallel control principle, 30 people used the samples continuously, and the initial value before using the samples and the body surface temperature 30 minutes after using the samples were calculated.

[0147] Test principle

[0148] Under the test conditions, the smaller the transepidermal water loss (TEWL) value, the less water is lost per unit time, that is, the better the skin barrier function.

[0149] Under the test conditions, the smaller the skin hemoglobin index, the less sensitive and red the skin is.

[0150] Under the test conditions, the lower the skin temperature value, the lower the skin temperature.

[0151] For sample information, refer to Table 10;

[0152] Table 10

[0153]

[0154] Subjects: A total of 30 subjects, 0 males and 30 females, aged 19 to 56 years, with an average age of 39±10 years, met the voluntary inclusion criteria.

[0155] Sample group: The sample group (control group + G-repair Xinyuening 2%) was used.

[0156] Control group: Use control group samples.

[0157] Test environment temperature: 20.0°C ~ 22.0°C; humidity: 40% ~ 60%, and dynamic monitoring.

[0158] Test method:

[0159] During the first visit, the subjects were given an explanation of the experiment and signed an informed consent form.

[0160] The subjects participating in the trial were screened according to the test requirements, and 30 people entered the experimental phase. The subjects cleaned their faces on the day of the test and equilibrated in the test environment for 20 minutes. The observed parts of the subjects were measured by transepidermal water loss meter, Antera 3D imager, and skin pH temperature meter.

[0161] After the measurement was completed, instructions on sample use were given and the samples were used. The sample group samples were used for the right side of the face, and the control group samples were used for the left side of the face.

[0162] A transepidermal water loss tester, Antera 3D imager, and skin pH and temperature measuring instrument were used to take photos and measure the skin at designated follow-up time points.

[0163] Result Analysis

[0164] The principle of before-after comparison

[0165] Descriptive statistics, including number, mean, and standard deviation, were performed on the measured values for each test area. The initial value and the measured values at other time points were calculated for each test area, and the differences between the measured values at different times and the initial value were statistically analyzed. If the test data showed a normal distribution, statistical analysis was performed using a rank sum test; if the test data showed a non-normal distribution, statistical analysis was performed using a rank sum test.

[0166] The statistical methods used were two-tailed tests with a test level of α-0.05.

[0167] Parallel control principle

[0168] Descriptive statistics were performed on the measurement values of each test area, including quantity, mean, and standard deviation.

[0169] The difference between the initial value of each test area and the measured value at other time points was calculated, and then this difference was used to statistically analyze the differences between the sample group and the control group at different time points.

[0170] If the test data is normally distributed, the test method is used for statistical analysis; if the test data is non-normally distributed, the rank sum test method is used for statistical analysis.

[0171] The statistical methods used were two-tailed tests, with a test level of a = 0.05.

[0172] Test results

[0173] Sample completion status

[0174] A total of 31 people were enrolled, 30 of whom completed the test and 30 were included in the statistical analysis.

[0175] Product usage and tolerance feedback

[0176] Each subject used the test sample as required within the specified time and no adverse reactions were observed.

[0177] Instrument test results and statistical analysis results

[0178] The transepidermal water loss (TEWL) value test results refer to Table 11;

[0179] Table 11

[0180]

[0181] Compared with before use, the subjects' transepidermal water loss (TEWL) values decreased significantly 30 minutes after using the sample, with a downward adjustment rate of 19.65%.

[0182] Compared with before use, after using the control group sample for 30 minutes, the transepidermal water loss (TEWL) value of the subjects' wrinkles decreased by 5.52%.

[0183] The results of skin hemoglobin index test are shown in Table 12;

[0184] Table 12

[0185]

[0186] Compared with before use, 30 minutes after using the sample, the subjects' skin hemoglobin index decreased significantly, with a decrease rate of 10.17%.

[0187] Compared with before use, the skin hemoglobin index of the subjects decreased by 2.94% 30 minutes after using the control group sample.

[0188] The results of the surface temperature test are shown in Table 13;

[0189] Table 13

[0190]

[0191] Compared with before use, the subjects' body surface temperature dropped significantly 30 minutes after using the sample, with a decrease rate of 4.29%.

[0192] Compared with before use, there was no improvement in the subjects' body surface temperature 30 minutes after using the control group samples.

[0193] Compared with the control group, the subjects' body surface temperature dropped significantly 30 minutes after using the sample.

[0194] in conclusion

[0195] After using the sample (sample group (control group + G-repair Xinyuening 2%)) for 30 minutes, the subjects' transepidermal water loss (TEWL) values were significantly decreased compared with before use, with a decrease rate of 19.65%. Compared with the control group, the subjects' transepidermal water loss (TEWL) values were significantly decreased.

[0196] 30 minutes after using the sample (sample group (control group + G-repair Xinyuening 2%)), the subjects' skin hemoglobin index decreased significantly compared with before use, with a decrease rate of 10.17%. Compared with the control group, the subjects' skin hemoglobin index decreased significantly.

[0197] 30 minutes after using the sample (sample group (control group + G-repair Xinyuening 2%)), the subject's body surface temperature decreased significantly compared with before use, with a decrease rate of 4.29%. Compared with the control group, the subject's body surface temperature decreased significantly.

[0198] Histamine model experiment

[0199] Subjects and number of participants: 3 adult volunteers aged 20 to 60 years, without skin diseases;

[0200] Test site: the inner side of the front end of the arm. Each volunteer tested the samples of Example 1 and Comparative Example 1, with one sample tested on each of the left and right arms.

[0201] Environmental conditions: Temperature: 21±1°C, Humidity: 50±10%;

[0202] Test method (puncture method): Use a puncture needle to puncture the test area on the inside of the arm, and the standard is no bleeding; use 1% histamine phosphate solution, drip 0.02ml on the inside of the arm, absorb the histamine solution after 5 minutes, and wait about 10 minutes. Apply 0.02g of sample to the sample area and the control area 2*2cm area, record the redness and itching of the arm at each time point, and score and record. The technician evaluates the test area at the corresponding time points (0min, 1min, 5min, 30min, 2h). The test results are shown in Table 14.

[0203] The improvement rate of histamine redness was calculated by collecting the scoring data of three volunteers and averaging them to one decimal place;

[0204] Histamine redness improvement rate (%) = (redness score at 0 min after histamine prick modeling - redness score at 2 h after histamine prick modeling) × 100% / redness score at 0 min after histamine prick modeling.

[0205] Table 14 Test results

[0206] blank 0min 1min 5min 30min 2h Improvement rate Comparative Example 1 4.5 3.2 2.7 2.3 2.1 53.33％ Example 1 4.8 3.4 3.2 2.0 1.7 64.58％

[0207] A test picture of one of the volunteers can be seen Figure 4 ;

[0208] The above tests show that the redness-removing effect of Example 1 of the present invention is significantly better than that of Comparative Example 1.

Claims

1. An extraction process for a Gentiana macrophylla extract, characterized in that: The steps include: Step 1: crushing the Gentiana macrophylla and dispersing it in water, and performing enzymolysis with cellulase to obtain an enzymolysis product; Step 2: Filter the enzymatic hydrolysis product to collect the filtrate and filter residue, and dry the filter residue to obtain a micropowder; after the filtrate is concentrated under reduced pressure and precipitated with alcohol, the first extract is collected; Step 3: extract the micropowder with alcohol once or multiple times, combine the extracts, and concentrate the extracts to obtain a thick paste; Step 4: Drying and powdering the thick paste to obtain a second extract; Step 5: Combine the first extract and the second extract to obtain the Gentiana macrophylla extract.

2. The extraction process of the Gentiana macrophylla extract according to claim 1, wherein In step 3, 95 vol% ethanol is used for the alcohol extraction, and the weight-to-volume ratio of the micropowder to the ethanol is 1:10 g / ml.

3. The extraction process of the Gentiana macrophylla extract according to claim 2, wherein: The number of alcohol extraction operations is 2 to 3 times, the single alcohol extraction time is 1 to 2 hours, and the alcohol extraction temperature is 50 to 65°C.

4. The extraction process of the Gentiana macrophylla extract according to claim 1, wherein In the step 1, the gentiana macrophylla is crushed and passed through a 300-mesh sieve, and then mixed with water in a mass-to-volume ratio of 1:5-10, and 0.1-0.2 wt% of cellulase is added to perform enzymolysis based on the weight of the gentiana macrophylla; The enzymatic hydrolysis time is 2 to 3 hours, and the enzymatic hydrolysis temperature is 45 to 65°C.

5. A Gentiana macrophylla extract, characterized in that: The method is prepared by any one of claims 1 to 4.

6. Use of the Gentiana macrophylla extract according to claim 5 in preparing cosmetics; the cosmetics are cosmetics that can soothe and / or alleviate allergies after acting on the skin.

7. A cosmetic, characterized in that: Contains the Gentiana macrophylla extract as claimed in claim 5.

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