CN117653580B - Herba Lophatheri extract, and its preparation method and application in cosmetics - Google Patents

Abstract

The present invention discloses a bamboo orchid extract, a preparation method thereof and an application thereof in cosmetics, and belongs to the field of traditional Chinese medicine. The bamboo orchid extract obtained by the present invention contains arbutin, beishengmanin, isoliquiritigenin and other substances involved in tyrosinase inhibition, as well as kaempferol-3-O-a-L-rhamnoside, luteolin, kaempferol, chlorogenic acid and other substances involved in anti-oxidation. The bamboo orchid extract can inhibit tyrosinase, and has antioxidant and moisturizing abilities, and can be used as a raw material for whitening, moisturizing and antioxidant cosmetics, and can effectively achieve whitening, freckle removal, prevention of melanin precipitation, moisturizing, anti-oxidation and other effects. No records of bamboo orchid extract being used in cosmetics have been detected in the prior art, and the application of bamboo orchid extract in cosmetics is helpful to increase the application scope and economic value of bamboo orchid extract.

Description

A kind of bamboo leaf orchid extract and its preparation method and application in cosmetics

Technical Field

The invention belongs to the field of traditional Chinese medicine, and particularly relates to a bamboo orchid extract and a preparation method thereof and application in cosmetics, specifically to a bamboo orchid extract and a preparation method thereof and whitening, anti-oxidation and moisturizing cosmetics added with the bamboo orchid extract.

Background Art

Arundina graminifolia is a plant of the genus Arundina of the Orchidaceae family. It is also known as grass ginger, mountain water chestnut, and Baiyangjie. It is a terrestrial herb. According to the Flora of China, the plant height of Arundina graminifolia is 40 to 80 cm. The flowering and fruiting period is mainly from September to November, but there are also periods from January to April. Arundina graminifolia is widely distributed in tropical and subtropical areas, such as Guangdong, Yunnan, Guangxi, Hainan, Tibet, etc. In the Dai area of Yunnan, Arundina graminifolia is a commonly used Dai medicine. Usually the whole plant or rhizome is used as medicine to treat food poisoning, snake bites, sores, boils, and swellings. It has the effects of regulating qi and blood, clearing heat and detoxifying. Studies in recent years have shown that Arundina graminifolia contains a large amount of stilbenes, flavonoids, esters, phenylpropanoids, etc. In addition, there are sterols, alkanes, phenolic acids and triterpenes. Some studies have shown that Arundina graminifolia extracts have antioxidant, anti-lipid peroxidation, anti-tumor, anti-pulmonary fibrosis, anti-viral and antibacterial activities.

At present, patents on the use of orchid plant extracts in skin care products or cosmetics (such as U.S. Patent US20110002968A1, Chinese invention patents CN107961195A and CN107028846A) are mostly used in the use of aerial orchids such as Phalaenopsis and Vanda, while the skin care use of terrestrial orchids with medicinal value such as Bambusa in bamboo leaves needs further development and utilization.

Summary of the invention

In order to overcome the shortcomings and deficiencies of the prior art, the object of the present invention is to provide a method for preparing a Bamboo Leaf Orchid extract.

Another object of the present invention is to provide a Bamboo Leaf Orchid extract prepared by the above preparation method. The extract has antioxidant ability, tyrosinase inhibition ability, and moisturizing ability, thereby achieving the purpose of improving skin care effects.

Another object of the present invention is to provide an application of the above-mentioned Bamboo Leaf Orchid extract. The Bamboo Leaf Orchid extract obtained by the present invention is suitable for adding into various cosmetics.

When analyzing the bamboo orchid extract, the present invention found that it contained a large number of important metabolites with whitening and antioxidant effects, but there was no record of the use of bamboo orchid extract in cosmetics in the prior art. The use of bamboo orchid extract in cosmetics will help to increase the application scope and economic value of bamboo orchid extract.

The purpose of the present invention is achieved through the following technical solutions:

A method for preparing a bamboo leaf orchid extract comprises the following steps:

(1) Pretreatment: Wash the bamboo orchid, cut it into pieces, and dry it; crush the dried bamboo orchid, and sieve it to obtain bamboo orchid powder;

(2) Treatment: soaking the powder of Bamboo Leaf Orchid with 10-50 mL of ethanol aqueous solution as the extraction ratio; wherein the volume fraction of the ethanol aqueous solution solvent is 0-80%; after the soaking, ultrasonic assisted extraction is performed, and the ultrasonic wave is repeated several times, each time for a certain period of time;

(3) separation: the extract obtained in step (2) is centrifuged or filtered to separate the feed and liquid, and the extracts are combined;

(4) Concentration and freeze-drying: The extract obtained in step (3) is concentrated by rotary evaporation to obtain an extract concentrate; the concentrate is freeze-dried to obtain a bamboo leaf orchid extract.

Preferably, in step (1), the Bamboo Cymbidium is the whole plant of Bamboo Cymbidium.

Preferably, in step (1), the cutting is cutting into pieces of 2 to 3 cm;

Preferably, in step (1), the sieving is through a 60-100 mesh sieve; further, through a 60 mesh sieve;

Preferably, in step (2), the extraction ratio is 1g of Bamboo Cymbidium powder: 30-50mL of ethanol-water solution solvent; further, the extraction ratio is 1g of Bamboo Cymbidium powder: 30-40mL of ethanol-water solution solvent; further, the extraction ratio is 1g of Bamboo Cymbidium powder: 40mL of ethanol-water solution solvent.

Preferably, in step (2), the volume fraction of the ethanol aqueous solution solvent is 20 to 80%; further 40 to 80%; further 60 to 80%; further 80%;

Preferably, in step (2), the soaking time is 2 to 6 hours; further, 2 hours.

Preferably, in step (2), the power of the ultrasonic-assisted extraction is 160 to 400 W; further 400 W;

Preferably, in step (2), the ultrasonication is performed 3 times;

Preferably, in step (2), each time the certain period of time is 10 to 50 minutes; further 30 to 50 minutes; further 40 minutes.

Preferably, in step (4), the freeze-drying time is 24 to 72 hours; further 48 hours.

A bamboo orchid extract is prepared by the above preparation method.

The bamboo orchid extract obtained by the invention contains arbutin, arbutin, isoliquiritigenin and other substances involved in tyrosinase inhibition, as well as kaempferol-3-O-a-L-rhamnoside, luteolin, kaempferol, chlorogenic acid and other substances involved in anti-oxidation.

The bamboo orchid extract can inhibit tyrosinase, effectively achieve the effects of whitening, removing spots, preventing melanin deposition, etc., and also has moisturizing and anti-oxidation capabilities;

The invention discloses an application of a bamboo orchid extract in preparing cosmetics.

The cosmetic contains the above-mentioned bamboo orchid extract, and the added amount in the cosmetic is 10-100 mg/L.

The cosmetics include: water-based lotions, emulsions, creams, sprays, essences, essential oils, soaps, cleansing products, makeup, etc.

Furthermore, the cosmetic is toner, lipstick or soap.

A bamboo orchid cosmetic contains the bamboo orchid extract.

Preferably, the bamboo orchid cosmetic contains 10 to 100 mg/L of the bamboo orchid extract.

The bamboo orchid cosmetics include: water-based lotions, emulsions, creams, sprays, essences, essential oils, soaps, cleansing and care products, and makeup products.

The bamboo orchid extract is mixed with cosmetic raw materials to obtain a bamboo orchid cosmetic. The cosmetic raw materials are a powder raw material, a paste raw material, and a liquid raw material. The bamboo orchid extract added is an extract powder, an extract aqueous solution, and an extract ethanol solution.

Specifically, the bamboo orchid cosmetic is a bamboo orchid toner, which contains the above-mentioned 10-100 mg/L bamboo orchid extract.

Specifically, the bamboo orchid cosmetic is a bamboo orchid lipstick, which contains 10 to 100 mg/L of the bamboo orchid extract.

Specifically, the bamboo orchid cosmetic is bamboo orchid soap, which contains the above-mentioned 10-100 mg/L bamboo orchid extract.

Compared with the prior art, the present invention has the following advantages and effects:

The bamboo orchid extract prepared by the present invention has an inhibitory effect on tyrosinase, and has antioxidant and moisturizing abilities, and can be used as a raw material for cosmetics with whitening, moisturizing and antioxidant effects. Arbutin, arbutin, isoliquiritigenin and other substances involved in tyrosinase inhibition, as well as kaempferol-3-O-a-L-rhamnoside, luteolin, kaempferol, chlorogenic acid and other substances involved in antioxidant effects are detected in the bamboo orchid extract.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a flow chart of the extraction method of the Bamboo Leaf Orchid extract.

FIG. 2 is a diagram showing the steps of preparing bamboo orchid cosmetics using bamboo orchid extract as an additive.

FIG3 shows the effects of different ethanol concentrations in the ethanol aqueous solution on the efficacy of the Bamboo Herb extract in step 2 of Example 1. Different lowercase letters represent significant differences between different treatments.

FIG. 4 shows the effects of different solid-liquid ratios on the efficacy of the Bamboo Herb extract in step 2 of Example 1. Different lowercase letters represent significant differences between different treatments.

FIG. 5 shows the effects of different ultrasonic extraction time treatments on the efficacy of the Bamboo Herb extract in step 2 of Example 1. Different lowercase letters represent significant differences between different treatments.

Figure 6 is a graph showing the difference in efficacy between the Bamboo Herb extract and the control extract (decoction in water for 1 hour or 2 hours) in Example 2; wherein different capital letters indicate significant differences in the same index between extracts obtained by different extraction methods, and different lowercase letters indicate significant differences in different indexes of the same extract.

Figure 7 is an appearance picture of the toner, lipstick and soap to which the bamboo orchid extract in Example 2 is added; wherein A: bamboo orchid toner, B: base toner, C: bamboo orchid soap, D: base soap, E: bamboo orchid lipstick, and F: base lipstick.

Figure 8 is an analysis chart of the efficacy of toners with the addition of the Bambusa orchid extract in Example 2; wherein different capital letters indicate significant differences in the same efficacy index between different toner groups, and different lowercase letters indicate significant differences in different efficacy indexes in the same toner group.

FIG. 9 is an analysis chart of the moisturizing ability of the toner and lipstick to which the Bamboo Herb extract of Example 2 is added on the skin of the test subjects after use.

FIG. 10 is an analysis of the relative change rate of skin pigmentation of subjects after using the toner containing the Bamboo Herb extract of Example 2.

DETAILED DESCRIPTION

The present invention is further described in detail below in conjunction with embodiments and drawings, but the embodiments of the present invention are not limited thereto.

The method for extracting bamboo orchid extract of the preferred embodiment of the present invention is applicable to the extraction of the whole bamboo orchid plant and its various parts (such as roots, leaves, flowers, etc.), and the method for adding bamboo orchid extract to skin care products is applicable to toner, face cream, lipstick, soap, etc.

Generally speaking, bamboo orchid contains a large amount of flavonoids and polyphenols, which have good antioxidant capacity. Reducing the generation of free radicals can delay aging. At the same time, the rhizome of bamboo orchid contains a large amount of phenanthrene substances, among which dihydrophenanthrene has a strong anti-tumor ability. The polysaccharides and protein peptides in plants have a particularly significant moisturizing effect on the skin. Some phenolic acids and flavonoids can reduce the production of melanin by inhibiting tyrosinase, thereby achieving a whitening effect. Therefore, bamboo orchid extract has a high potential medicinal and economic value. In addition, bamboo orchid extract has no special smell and is suitable as an additive.

FIG. 1 is a flow chart of the extraction method of the Bamboo Leaf Orchid extract.

1. Pretreatment: Wash and dry the whole plant of Bamboo Leaf Orchid, cut it into pieces of 2-3 cm, spread it flat and dry it. Crush the dried Bamboo Leaf Orchid and sieve it through 60 mesh to obtain Bamboo Leaf Orchid powder.

2. Extraction: Following the previous step, take the bamboo orchid powder and add ethanol aqueous solution solvent, soak for 2 hours. The volume fraction of ethanol aqueous solution solvent is 0-80%, and 10-50 mL of solvent is added for each gram of bamboo orchid.

After soaking, the extract is placed in an ultrasonic instrument for ultrasonic-assisted extraction. The ultrasonic process is repeated 3 times, each time for 10 to 50 minutes. The ultrasonic extraction power is 160 to 400W.

3. Separation: The separation step in Figure 1 refers to separating the extract obtained in the previous step by passing it through a centrifuge or filter paper to separate the feed liquid and combine the extracts.

4. Concentration: As shown in FIG1 , the extract obtained in the previous step is subjected to a rotary evaporator to remove the solvent and concentrate to obtain an extract concentrate.

5. Freeze drying: Freeze drying the concentrate means freezing the extract concentrate into ice cubes and placing it in a freeze dryer for 24 to 72 hours to obtain the Bamboo Leaf Orchid extract.

The preferred bamboo orchid extract obtained according to the above steps can be used as an additive to prepare bamboo orchid cosmetics. As shown in Figure 2, the cosmetic raw materials can be in powder, paste, or liquid form, and the added bamboo orchid extract is extract powder and/or extract polar solution (such as ethanol) and/or extract aqueous solution.

Embodiment 1

1. Randomly set the extraction method of bamboo leaf orchid extract: add 3g bamboo leaf orchid powder to 60mL 60% ethanol aqueous solution solvent (water: ethanol = 4: 6) (i.e., material-liquid ratio (g: mL) = 1: 20), and let it stand and soak for 2h. Put it into an ultrasonic instrument, extract it with 400W power for 30min, and centrifuge to get the supernatant. Repeat the ultrasonic extraction three times and combine the extracts. Concentrate the extract by rotary evaporation to obtain the extract concentrate, and freeze-dry the concentrate for 48h to obtain the bamboo leaf orchid extract.

2. Using step 1 as the extraction method, reveal the effects of single factor treatments: different ethanol concentrations in ethanol aqueous solution (0%, 20%, 40%, 60%, 80%), different solid-liquid ratios (1:10, 1:20, 1:30, 1:40, 1:50), and different ultrasonic extraction times (10min, 20min, 30min, 40min, 50min) on the efficacy of bamboo orchid extract (tyrosinase inhibition rate, DPPH free radical scavenging rate, ABTS ⁺ free radical scavenging rate).

3. Tyrosinase inhibition rate: Prepare a sample solution with a concentration of 1 mg/mL, add phosphate buffer (pH 6.8, weigh 8.95 g NaH ₂ PO ₄ ·2H ₂ O and 3.90 g Na ₂ HPO ₄ ·12H ₂ O with a 1/1000 scale, add them to 500 mL of deionized water, stir evenly with a glass rod to obtain PBS buffer), sample solution to be tested, tyrosinase solution (150 U/mL), mix well and place at 37°C for activation reaction for 10 min, then add L-tyrosine solution (0.3 mg/mL) and react for 20 min in the dark. Use an enzyme marker to measure the absorbance of each well at a wavelength of 475 nm. The sample is repeated three times, and the specific addition amount is shown in Table 1. Among them, the concentration of arbutin solution is 4 mg/mL.

Table 1 250μL experimental system

Note: When measuring absorbance, the “sample group”, “arbutin group” and “blank group” were adjusted to zero using the “sample control group”, “arbutin control group” and “blank control group” respectively.

Inhibition rate (%) = (A-B)/A×100%

Among them, A is the absorbance value of the blank group, and B is the absorbance value of the sample group to be tested.

4. DPPH free radical scavenging rate: prepare several 2mL centrifuge tubes, add 1mL of 0.5mg/mL sample solution and 1mL of DPPH solution (0.2mmol/L) and mix well, and react at 37℃ in the dark for 30min. Mix 1mL of anhydrous ethanol with 1mL of sample solution for sample zero adjustment, and mix 60% ethanol with DPPH solution as blank group. Repeat three times for each sample, and measure the absorbance value at a wavelength of 519nm.

The scavenging rate of the sample on DPPH free radicals (%) = [1-A/A0] × 100%

Where A is the absorbance value of the sample group to be tested; A0 is the absorbance value of the blank group.

5. ABTS ⁺ free radical scavenging rate: Take the sample solution with a concentration of 0.125 mg/mL, add the sample solution and ABTS ⁺ solution (2.6 mmol/L) into the test tube in a volume ratio of 1:2, mix well, place in the dark for 6 minutes, and measure the absorbance at a wavelength of 734 nm. Use anhydrous ethanol instead of the sample solution as a blank control, and repeat three times for each concentration. The ABTS ⁺ free radical scavenging rate is calculated using the following formula:

ABTS ⁺ free radical scavenging rate (%) = (1-A1/A) × 100%

A: absorbance of blank control; A1: absorbance of the sample group to be tested.

6. The experimental results are shown in Figures 3 to 5.

As shown in Figure 3, when the ethanol concentration is between 0 and 80%, the tyrosinase inhibition rate of the extract gradually increases with the increase of ethanol concentration, and the tyrosinase inhibition rate is the highest at 80% ethanol concentration, reaching 56.6%. As the ethanol concentration of the extract increases, the DPPH free radical scavenging rate of the extract sample gradually increases, and the scavenging rate reaches 82.9% when the ethanol concentration reaches 80%. At the same time, as the ethanol concentration increases, the ABTS ⁺ free radical scavenging rate of the sample also gradually increases, and the scavenging rate reaches 78.64% at 80%. In summary, the three indicators of the bamboo leaf orchid extract show an upward trend with the increase of ethanol concentration, and 80% is the optimal choice for ethanol concentration.

As shown in Figure 4, in the single-factor experiment of the material-liquid ratio, it was found that the tyrosinase inhibition rate reached a peak at 1:40g/mL and began to decline at 1:50g/mL. The tyrosinase inhibition rate reached 68.3% at a material-liquid ratio of 1:40g/mL. The DPPH free radical scavenging rate under different material-liquid ratios gradually decreased before 1:30g/mL, and showed an upward trend after exceeding 1:30, but there was little difference between the groups; the ABTS ⁺ free radical scavenging rate reached a maximum of 81.35% at 1:50. The effect of the material-liquid ratio on the free radical scavenging rate is not much different, but the effect on the tyrosinase inhibition rate is greater. When the material-liquid ratio is low, the extract cannot be completely precipitated, and too high a material-liquid ratio will cause waste of the extract, so 1:40g/mL is selected as the optimal material-liquid ratio.

As shown in Figure 5, in the single-factor test of ultrasound time, before 40 minutes, as the ultrasound time increased, the tyrosinase inhibition rate of the extract gradually increased, reaching a peak at 40 minutes, and then began to decline. The changes in the DPPH free radical scavenging rate and the ABTS ⁺ free radical scavenging rate were not much different, so 40 minutes was selected as the optimal ultrasound time.

7. Through single factor experiments, the optimal process combination for bamboo orchid extract was determined to be: ethanol concentration 80%, solid-liquid ratio 1:40g/mL, ultrasonic time 40min, and ultrasonic power 400W.

Embodiment 2

The components of Bamboo Spathiphyllum extract were identified and relatively quantitatively analyzed by UPLC-MS/MS method.

1. Add 3g of Bamboo Leaf Orchid powder to 120mL of ultrapure water, heat to boiling and start timing, keep boiling for 1h. Concentrate by rotary evaporation to obtain extract concentrate, freeze-dry the concentrate for 48h to obtain Bamboo Leaf Orchid extract. Recorded as 1h decoction group (abbreviation: ZY1).

2. Add 3g of Bamboo Leaf Orchid powder to 120mL of ultrapure water, heat to boiling and start timing, keep boiling for 2h. Concentrate by rotary evaporation to obtain extract concentrate, freeze-dry the concentrate for 48h to obtain Bamboo Leaf Orchid extract. Recorded as 2h decoction group (abbreviation: ZY2).

3. Using the best process for the extract of Bamboo Leaf Orchid in Example 1, add 3g Bamboo Leaf Orchid powder to 120mL 80% ethanol aqueous solution solvent (water: ethanol = 2: 8) (i.e., solid-liquid ratio (g: mL) = 1: 40), and let it stand and soak for 2h. Put it into an ultrasonic instrument, extract it with 400W power for 40min, and centrifuge to take the supernatant. Repeat the ultrasonic extraction three times and combine the extracts. Concentrate the extract by rotary evaporation to obtain the extract concentrate, freeze-dry the concentrate for 48h to obtain the Bamboo Leaf Orchid extract. Recorded as ultrasonic alcohol extraction group (abbreviation: ZY3).

4. The extracts obtained by different extraction methods of Bamboo Leaf Orchid were tested for DPPH free radical scavenging rate, ABTS ⁺ free radical scavenging rate, and tyrosinase inhibition rate. The results are shown in Figure 6. It can be seen that the DPPH free radical scavenging rate and ABTS ⁺ free radical scavenging rate of the water decoction method increased significantly with the increase of extraction time, but the tyrosinase inhibition rates were 64.3% and 65.7%, respectively, and there was no significant difference between the two. The free radical scavenging rate and tyrosinase inhibition rate of the ultrasonic-assisted alcohol extraction method were greatly improved compared with the water decoction method, with a DPPH free radical scavenging rate of 89.8%, an ABTS ⁺ free radical scavenging rate of 84.9%, and a tyrosinase inhibition rate of 83.4%. The results showed that during the water decoction extraction process, the antioxidant activity of Bamboo Leaf Orchid increased with time, while the whitening ability remained basically unchanged. The extract obtained by ultrasonic-assisted alcohol extraction of Bamboo Leaf Orchid has higher antioxidant activity and whitening activity than the extract obtained by water decoction. The extracts of the bamboo leaf orchid extracted by water extraction for 1 hour and 2 hours were used as controls, and the UPLC-MS/MS method was used to identify and relatively quantitatively analyze the components of the bamboo leaf orchid extract obtained by the optimal process of the bamboo leaf orchid extract in Example 1.

5. UPLC-MS/MS method

(1) Sample processing: The vacuum freeze-dried sample powder was crushed into powder using a grinder at 30 Hz for 1.5 min; 100 mg of the powder was weighed and dissolved in 1.0 mL of 70% methanol extract; the sample was vortexed once every 30 min for 30 s each time, for a total of 6 times, and the sample was placed in a 4°C refrigerator overnight; the sample was centrifuged at 12,000 rpm for 10 min, the supernatant was aspirated, the sample was filtered with a microporous filter membrane (0.22 μm pore size), and stored in an injection bottle for UPLC-MS/MS analysis.

(2) Chromatographic mass spectrometry acquisition conditions

The data acquisition instrument system mainly includes ultra-high performance liquid chromatography (UPLC) (Shim-pack UFLC SHIMADZU CBM30A, https://www.shimadzu.com/) and tandem mass spectrometry (MS/MS) ( 4500+, https://sciex.com/).

The liquid phase conditions mainly include: chromatographic column: Waters ACQUITY UPLC HSS T3 C18 1.8μm, 2.1mm*100mm; mobile phase: phase A is ultrapure water (0.1% formic acid), phase B is acetonitrile (0.1% formic acid); elution gradient: 0min water/acetonitrile (95:5V/V), 10.0min is 5:95V/V, 11.0min is 5:95V/V, 11.1min is 95:5V/V, 15.0min is 95:5V/V; flow rate is 0.4ml/min; column temperature is 40℃; injection volume is 2μl.

The mass spectrometry conditions mainly include: electrospray ionization (ESI) temperature 550°C, mass spectrometry voltage 5500V (positive), -4500V (negative), ion source gas I (GS I) 55psi, gas II (GS II) 60psi, curtain gas (CUR) 25psi, collision-activated dissociation (CAD) parameters set to high. In the triple quadrupole (Qtrap), each ion pair is scanned and detected according to the optimized declustering potential (DP) and collision energy (CE).

6. By comparing with the local metabolic database and sample metabolites, a total of 669 metabolites were identified in this experiment, 400 metabolites were detected in the positive ion mode, and 269 metabolites were detected in the negative ion mode. Through correlation analysis, some differential substances highly correlated with efficacy indicators were obtained, as shown in Table 2. These substances have been shown to have antibacterial, tyrosinase inhibition and antioxidant effects in previous studies. Arbutin, salicylic acid, and ferulic acid have high skin care effects and have been added to skin care products as whitening agents in recent years. Isoliquiritigenin, naringenin, and butein are substances that appear in alcohol extraction extracts and have a very high positive correlation with efficacy indicators. Many scholars have found through research that substances such as isoliquiritigenin and naringenin in licorice have good tyrosinase inhibition ability, which can inhibit the binding of tyrosinase to substrates and thus reduce the formation of melanin. Astragaloside, kaempferol, chlorogenic acid, and quercetin are common flavonoid compounds with antioxidant capacity in plant extracts. These substances have also been detected in the extracts of Bamboo Orchid. The content of these substances is higher in the alcohol extraction extract, resulting in higher free radical scavenging ability.

Table 2 Some differential substances in Bamboo Leaf Orchid extracts that are highly correlated with efficacy indicators

Note: CAS is the CAS number of the substance, ZY1 represents the relative content of the component extracted by water extraction for 1 hour, ZY2 represents the relative content of the component extracted by water extraction for 2 hours, ZY3 represents the relative content of the component extracted by the best extraction process, and cpd_ID represents the ID number of the substance in the KEGG database.

Embodiment 3

1. To further explore the potential of bamboo orchid extract as a raw material for functional skin care products, toner, lipstick, and soap with bamboo orchid extract were prepared, and the quality of the samples was evaluated. The antioxidant, whitening, moisturizing, and antibacterial abilities of the samples were measured by comparing them with the matrix group without bamboo orchid extract to determine whether the skin care products with bamboo orchid extract have good safety and efficacy. The bamboo orchid extract obtained by the optimal process in Example 1 was used for subsequent experiments.

Among them, the preparation of bamboo orchid lipstick: the matrix group formula is selected to weigh olive oil and yellow beeswax in a ratio of 3:1 and put them into a clean beaker, put the beaker into a water bath and heat it at a constant temperature of 70°C until the yellow beeswax melts, and stir it evenly until there are no bubbles. The resulting oil is allowed to stand and cool to about 60°C, and 2% vitamin E and bamboo orchid extract solution (concentration of 1 mg/mL) 1% v/v are added and mixed; the first filling is to 90% of the mold, and the remaining oil is poured in after one minute. Finally, it is allowed to stand at room temperature for about 1 hour to form, and the bamboo orchid lipstick is obtained, as shown in Figure 7; wherein, the final concentration of bamboo orchid extract in the lipstick is 1% w/w.

Preparation of bamboo orchid soap: According to the formula of pure natural handmade soap, natural transparent soap base and 2% amino acid foaming agent are selected as the base. First, pour the soap base into a beaker, heat it at 80°C in water, and slowly stir to increase the melting speed until it is completely melted into liquid and taken out. Add 2% amino acid foaming agent and 1% v/v bamboo orchid extract solution (concentration of 1 mg/mL), stir slowly with a spatula, and finally use a spatula to block the bubbles floating on the upper layer in a warm state and slowly pour the soap solution into the mold. Wait for 1-2 hours and then demold to obtain the finished soap, and prepare bamboo orchid soap, as shown in Figure 7; wherein, the final concentration of bamboo orchid extract in the soap is 10 mg/L.

Preparation of bamboo orchid toner: prepare deionized water, glycerin, hyaluronic acid, and vitamin B3 as toner base materials, and adjust the toner formula to include 72% deionized water, 10% glycerin, 6% hyaluronic acid, 2% vitamin _B3 , and 10% bamboo orchid extract solution (concentration of 1 mg/mL). First, mix glycerin and deionized water in proportion, then add hyaluronic acid, vitamin _B3 , and bamboo orchid extract solution in sequence, and shake thoroughly until homogeneous to obtain bamboo orchid toner, as shown in Figure 7; wherein, the final concentration of bamboo orchid extract in the toner is 100 mg/L.

2. Quality evaluation of skin care products containing Bamboo Orchid extract:

(1) Sensory evaluation: The sensory indicators of bamboo orchid lipstick, toner, and soap were evaluated according to the national standard GB/T 26513-2011. The color, smell, and shape of the skin care products containing bamboo orchid extract were observed; whether the surface of the lipstick and soap was smooth, whether there were pores and impurities, etc.; and whether the texture of the toner was uniform.

(2) Cold resistance test: Take 2 skin care products (two lipsticks, 2 bars of soap or 2 toners), keep one as a control at room temperature, take out the experimental skin care product and place it in a -10℃ refrigerator for 24 hours. After taking it out and returning it to room temperature, observe whether its shape is bent or layered, whether there are cracks on the surface of the lipstick or soap, and whether the application experience changes.

(3) Heat resistance test: Take two lipsticks, two bars of soap or two portions of toner, twist out the lipstick, and place the soap or toner in a constant temperature box at room temperature and 45°C respectively. After 24 hours, take out the skin care products in the constant temperature box. After they return to room temperature, compare the shapes of the two skin care products to see if they are bent, softened or layered, and smell whether the lipstick has a spoiled odor.

(4) Stability test: Place the bamboo orchid toner in a centrifuge tube and centrifuge it at 3750 rpm for 30 min (equivalent to one year's gravity). Observe whether its texture is uniform and whether there is any stratification to evaluate the stability of the toner.

3. The results of the evaluation of the quality of bamboo orchid extract skin care products showed that the bamboo orchid extract solution (concentration of 1 mg/mL) was yellow-green, and the color of the skin care products gradually deepened with the increase of the amount added. When the amount added was 20% v/v, each skin care product was dark yellow-green, which did not meet the sensory requirements and had a certain impact on stability. Comprehensive consideration was given to ensure that the sensory organs were within the appropriate range and to ensure a certain efficacy, so the amount of bamboo orchid extract solution added was 1-10% as the final formula. The prepared bamboo orchid lipstick, soap, and toner were shown in Figure 7. The quality of the prepared bamboo orchid skin care products was evaluated, and the results are shown in Table 3. Compared with the control group without bamboo orchid extract, the addition of bamboo orchid extract did not have a significant effect on the quality of the skin care products. After adding bamboo orchid extract to the toner, the color changed from transparent and colorless to yellow-green, with a slight bamboo orchid fragrance, which had no effect on the skin feel of the toner, and the pH value was within the range acceptable to human skin and would not cause skin irritation. Since the lipstick uses yellow beeswax as its base, the color of the paste does not change significantly after adding bamboo orchid. The soap is transparent light yellow and has no special smell. The three bamboo orchid skin care products did not deteriorate or change in shape after the cold and heat resistance tests, indicating that the addition of bamboo orchid extract has no effect on the properties of the skin care products. Toner, as a skin care product that is a mixture of oil and water, did not show obvious stratification after gravity centrifugation equivalent to one year, indicating that it has good stability. In addition, all skin care products did not show oil-water separation or deterioration after being placed at room temperature for three months.

Table 3 Quality evaluation of bamboo leaf orchid skin care products

4. The tyrosinase inhibition rate, DPPH free radical scavenging rate, and ABTS ⁺ free radical scavenging rate test methods in Example 1 were used to test the tyrosinase inhibition rate, DPPH free radical scavenging rate, and ABTS ⁺ free radical scavenging rate of the bamboo orchid toner.

The results are shown in Figure 8. The antioxidant capacity and tyrosinase inhibition rate of the toner with bamboo orchid extract were measured. The toner with the same formula but without bamboo orchid extract was used as a control. The toner with bamboo orchid extract was significantly higher than the matrix group. The matrix group also has a certain antioxidant capacity due to the addition of matrix with antioxidant capacity such as vitamin _B3 , but there is still a significant difference compared with the bamboo orchid extract toner. There are also significant differences in the performance of the two toners in the tyrosinase inhibition rate, but the tyrosinase inhibition rate of the toner with bamboo orchid extract is only about 33%. This may be because the amount added is small and the content of tyrosinase inhibition cannot be reached. In summary, the bamboo orchid toner retains the antioxidant capacity and tyrosinase inhibition ability of bamboo orchid extract.

5. The ability to moisturize the skin is the most basic indicator of skin care products. Toners and lipsticks with excellent functions should have good moisturizing and hydrating abilities. In order to further explore the moisturizing and whitening abilities of bamboo orchid skin care products on human skin, the moisture content and colorimetry of human skin were measured after using bamboo orchid skin care products.

(1) Determination of skin moisture content: 10 volunteers of random gender and aged between 18 and 35 years old were randomly selected for the test. No cosmetics were allowed to be applied to the test area on the morning of the test. First, the volunteers washed the inner area of their arms with clean water and marked a 4cm×4cm experimental area. Then they sat quietly in the experimental room with a temperature of about 25°C and an air humidity of about 60% and waited for 30 minutes. Skin care products with bamboo orchid extract were applied to the experimental area. The same matrix skin care products without bamboo orchid extract were used as positive controls, and no skin care products were applied as negative controls. During the test, volunteers were not allowed to leave the test room, and they were not allowed to eat or engage in strenuous activities. The test cycle was 3 hours. The skin moisture content was measured using a skin tester every 30 minutes. The test was repeated three times for each area and the average value was taken.

(2) Skin pigmentation test: Skin pigmentation is tested using a skin analyzer based on the principle of spectral absorption. During the test, the probe is pressed vertically on the skin surface. After the test is completed, the subject's skin color value (0-99) can be viewed. The higher the value, the lower the skin melanin value.

(3) Randomly select 5 volunteers aged between 18 and 24 years old. Mark a 4 cm × 4 cm test area on the inner arm area of the subject and apply the product once in the morning and evening every day. Wash the test area before application. Do not do strenuous exercise within 30 minutes after application to ensure the absorption of the sample. Test the skin pigmentation of the volunteers on days 0, 7, 14, and 21, and repeat three times to take the average value.

The experimental results are shown in Figure 9. Taking the relative change rate before and after use at 30, 60, 90, 120, 150, and 180 minutes as an indicator, all samples did not cause irritation to the subjects' skin during the test period, and no redness, swelling, allergies, etc. were observed. Bamboo orchid toner and matrix toner both showed good hydration ability within 0-2 hours of use, and the skin moisture content was significantly improved compared to before use. At the 180th minute, the relative change rate of the bamboo orchid toner group was 22.2%, and that of the matrix group was 7.4%. Compared with the blank group that did not use toner, the toner showed good moisturizing ability. During the test, the relative change rate of the bamboo orchid group in the first 160 minutes changed more slowly than that of the matrix group, and at 180 minutes, the relative change rate was still significantly higher than that of the matrix group; the skin moisture change rate of the bamboo orchid lipstick was 3.9% after 120 minutes of use, which was higher than that of the matrix group; this shows that the skin care products added with bamboo orchid showed good hydration and moisturizing ability.

The subjects used the bamboo orchid toner for 0, 7, 14, 21, and 28 days, and the relative change rate of the skin pigmentation was calculated as shown in Figure 10. The base toner without bamboo orchid extract had a lower change rate compared with the initial value during the 7-28 days of use, and there was no significant difference in the skin pigmentation at each stage, indicating that the toner without bamboo orchid extract did not have a whitening effect. The skin pigmentation of the toner with bamboo orchid extract began to change significantly from 14 to 21 days of use, and the change rate of skin pigmentation reached 2.4% on the 28th day relative to the previous use. The skin pigmentation value showed a significant change relative to the initial value on the 28th day of use, indicating that the bamboo orchid toner has a certain whitening effect.

The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims (7)

1. The application of the herba Lophatheri extract in preparing cosmetics is characterized in that: the herba Lophatheri extract has inhibiting effect on tyrosinase, and can effectively achieve effects of whitening, removing freckle and preventing melanin precipitation, and meanwhile has moisturizing capability;

the preparation method of the herba phyllanthi extract comprises the following steps:

(1) Pretreatment: cleaning herba Lophatheri, cutting, and oven drying; pulverizing dried herba Lophatheri, and sieving to obtain herba Lophatheri powder;

(2) And (3) treatment: soaking the dried herba phyllanthi powder in 10-50 mL ethanol water solution solvent of 1 g; wherein the volume fraction of the ethanol aqueous solution solvent is 0-80%; carrying out ultrasonic auxiliary extraction after soaking, and carrying out ultrasonic treatment for a plurality of times, wherein each time lasts for a certain time;

(3) Separating: separating the liquid extract obtained in the step (2) through centrifugation or filtration, and combining the liquid extracts;

(4) Concentrating and freeze drying: concentrating the extract obtained in the step (3) through rotary evaporation to obtain an extract concentrated solution; freeze drying the concentrated solution to obtain the herba Lophatheri extract.

2. The use according to claim 1, characterized in that:

in the step (2), the power of the ultrasonic auxiliary extraction is 160-400W;

in the step (2), the duration of each time is 10-50 min.

3. The use according to claim 2, characterized in that:

in the step (2), 1g of bamboo She Lanfen powder 30-50 mL of ethanol water solution solvent is taken as an extraction proportion;

In the step (2), the volume fraction of the ethanol aqueous solution solvent is 20-80%;

in the step (2), the duration of each time is 30-50 min.

4. A use according to any one of claims 1 to 3, characterized in that:

In the step (1), the bamboo She Lanwei bamboo She Lanquan strain;

In the step (1), the shearing is carried out until the shearing is carried out to 2-3 cm;

In the step (1), the sieving is that a 60-100 mesh sieve is adopted;

In the step (2), the soaking time is 2-6 hours;

in the step (2), the ultrasonic frequency is 3 times;

in the step (4), the time of freeze drying is 24-72 h.

5. A use according to any one of claims 1 to 3, characterized in that:

The cosmetic contains the herba Lophatheri extract, and the addition amount of the herba Lophatheri extract in the cosmetic is 10-100 mg/L.

6. A use according to any one of claims 1 to 3, characterized in that:

the cosmetic comprises the following components: water, emulsion, cream, spray, essence, essential oil, soap, washing and caring or make-up.

7. The use according to claim 6, characterized in that:

the cosmetic is toner, lipstick or soap.