Cacumen biotae extract as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of plant extraction, and discloses a cacumen biotae extract as well as a preparation method and application thereof. The preparation method of the cacumen biotae extract comprises the following steps: mixing cacumen biotae and an extraction solvent, and performing high-pressure homogenization to obtain a cacumen biotae homogeneous solution; heating, extracting and filtering the Chinese arborvitae twig and leaf homogenized liquid to obtain a Chinese arborvitae twig and leaf extract; the extraction solvent comprises a eutectic solvent and water; the eutectic solvent comprises a hydrogen bond acceptor and a hydrogen bond donor, the hydrogen bond acceptor comprises urea and / or betaine, and the hydrogen bond donor comprises sorbitol and / or trehalose. According to the preparation method, the specific deep-eutectic solvent is selected, and a high-pressure homogenization green extraction process is matched, so that active ingredients in the cacumen biotae can be fully released and dissolved, the utilization efficiency of the raw materials is improved, and the prepared cacumen biotae extract contains high-content total flavonoids, myricetin, isoquercitrin, quercitrin, amentoflavone and quercetin; therefore, the traditional Chinese medicine composition has remarkable anti-oxidation and anti-inflammatory effects, and is mild and non-irritant.

Description

Technical Field

[0001] The invention belongs to the technical field of plant extraction, and particularly relates to a Platycladus orientalis leaf extract and a preparation method and application thereof. Background Art

[0002] Platycladus orientalis belongs to the Selaginellaceae family, and its extracts have been included in the 2021 edition of the "Catalogue of Used Cosmetic Raw Materials". According to reports, Platycladus orientalis leaves have a variety of flavonoids, mainly containing myricitrin, isoquercetin, quercetin and spicatelin. Pharmacological studies have shown that Platycladus orientalis leaf extracts have anti-inflammatory, antioxidant and antibacterial effects and are widely used in washing and care products. For the flavonoid components in Platycladus orientalis leaves, traditional extraction methods mainly rely on organic solvents such as ethanol, methanol and ethyl acetate; although these methods are effective to a certain extent, there are some problems, such as the low content of active ingredients in the extracts, solvent volatilization, and environmental pollution.

[0003] Therefore, how to extract flavonoids from Platycladus orientalis leaves in a green and efficient manner is a technical problem that needs to be solved urgently in this field. Summary of the invention

[0004] The present invention aims to solve at least one of the technical problems existing in the above-mentioned prior art. To this end, the present invention provides a Platycladus orientalis leaf extract and a preparation method and application thereof. The present invention realizes the green and efficient extraction of flavonoid components in Platycladus orientalis leaves by combining a low eutectic solvent with high-pressure homogenization. The obtained Platycladus orientalis leaf extract has a high content of total flavonoids and myricitrin, isoquercitrin, quercitrin, spicate biflavonoids, and quercetin, thereby having significant antioxidant and anti-inflammatory effects, and is mild and non-irritating.

[0005] In a first aspect, the present invention provides a method for preparing a Platycladus orientalis leaf extract, comprising the following steps:

[0006] The Platycladus orientalis leaves and the extraction solvent are mixed and homogenized under high pressure to obtain a Platycladus orientalis leaf homogenate;

[0007] The Platycladus orientalis leaf homogenate is subjected to heating extraction and filtering to obtain the Platycladus orientalis leaf extract;

[0008] The extraction solvent includes a deep eutectic solvent and water;

[0009] The deep eutectic solvent comprises a hydrogen bond acceptor and a hydrogen bond donor, wherein the hydrogen bond acceptor comprises urea and / or betaine, and the hydrogen bond donor comprises sorbitol and / or trehalose.

[0010] Specifically, deep eutectic solvents (DESs) are a mixture of hydrogen bond acceptors and hydrogen bond donors, which form a uniform transparent liquid under heating and stirring, have a low melting point, and excellent biocompatibility. The active ingredients in the plants will further form hydrogen bonds and van der Waals forces with the components in the deep eutectic solvents, introducing more binding forces into the system. This interaction not only does not destroy the internal structure of the deep eutectic solvent, but makes the structure of the system more stable, and helps the deep eutectic solvent to extract the active ingredients efficiently and more effectively and in a targeted manner. Deep eutectic solvents have the advantages of diverse structures, low prices, easy availability of raw materials, simple preparation, and no use of volatile organic solvents.

[0011] Compared with traditional organic solvent extraction, low eutectic solvents have the following advantages in the extraction of flavonoid components: 1) Efficient extraction: low eutectic solvents can form hydrogen bonds and other interactions with flavonoid components, thereby improving the extraction efficiency. 2) Low cost: The raw materials of low eutectic solvents are easy to obtain, simple to prepare, and relatively low in cost, which helps to reduce the cost of the extraction process and improve economic benefits. 3) Easy to recycle and reuse: low eutectic solvents are not easy to volatilize during the extraction process, and are easy to recycle and reuse, which helps to reduce resource waste and environmental pollution. 4) Green and environmentally friendly: low eutectic solvents do not use volatile organic solvents, which reduces the risk of environmental pollution and solvent volatilization, and is in line with the current concept of green chemistry and sustainable development.

[0012] Based on the 2021 edition of the "Catalogue of Used Cosmetic Raw Materials", the present invention has developed a low eutectic solvent suitable for use in the cosmetics field, and combined with a high-pressure homogenization process, it can efficiently extract flavonoid components in Platycladus orientalis leaves while being mild. This is an important technological breakthrough in the application of Platycladus orientalis leaf extract in the cosmetics field, and it meets the needs of green and high-quality development of cosmetics.

[0013] In some embodiments of the present invention, the usage ratio of the Platycladus orientalis leaves to the extraction solvent is 1g:(0.5-50g).

[0014] In some embodiments of the present invention, the usage ratio of the low eutectic solvent to the water is 1g:(0.05-20g).

[0015] In some embodiments of the present invention, the usage ratio of the hydrogen bond acceptor to the hydrogen bond donor is 1 g:(0.2-5 g).

[0016] In some embodiments of the present invention, the pressure of the high-pressure homogenization is 20-100 MPa; and / or, the temperature of the high-pressure homogenization is 25-50° C.; and / or, the time of the high-pressure homogenization is 5-40 min.

[0017] In some embodiments of the present invention, the temperature of the heating extraction is 90-110° C.; and / or the time of the heating extraction is 10-40 min.

[0018] The second aspect of the present invention provides a Platycladus orientalis leaf extract, which is prepared by the preparation method described in the first aspect of the present invention, and the content of total flavonoids in the Platycladus orientalis leaf extract is greater than 20 mg / g.

[0019] In some embodiments of the present invention, the content of myricitrin in the Platycladus orientalis leaf extract is >100 μg / g.

[0020] In some embodiments of the present invention, the content of isoquercetin in the Platycladus orientalis leaf extract is greater than 10 μg / g.

[0021] In some embodiments of the present invention, the content of quercetin in the Platycladus orientalis leaf extract is greater than 2230 μg / g.

[0022] In some embodiments of the present invention, the content of flavonoids in the Platycladus orientalis leaf extract is greater than 40 μg / g.

[0023] In some embodiments of the present invention, the content of quercetin in the Platycladus orientalis leaf extract is greater than 7 μg / g.

[0024] The third aspect of the present invention provides a cosmetic comprising the Platycladus orientalis leaf extract according to the first aspect of the present invention.

[0025] In some embodiments of the present invention, the content of the Platycladus orientalis leaf extract in the cosmetic is 0.01 wt %-50 wt %.

[0026] In some embodiments of the present invention, the content of the Platycladus orientalis leaf extract in the cosmetic is 0.1 wt %-30 wt %.

[0027] In some embodiments of the present invention, the content of the Platycladus orientalis leaf extract in the cosmetic is 0.1 wt %-10 wt %.

[0028] In some embodiments of the present invention, the cosmetic is selected from any one of shampoo, conditioner, hair oil, shower gel, facial cleanser, facial mask, lotion, facial cream or essence.

[0029] In some embodiments of the present invention, the cosmetic is a shampoo, and the shampoo further comprises at least one of a surfactant, a film former, a chelating agent, a pH adjuster, and a preservative.

[0030] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0031] The present invention selects a specific low eutectic solvent and cooperates with a high-pressure homogenization green extraction process, so that the active ingredients in the Platycladus orientalis leaves can be fully released and dissolved, and the utilization efficiency of the raw materials can be improved. The obtained Platycladus orientalis leaf extract has a high content of total flavonoids and myricitrin, isoquercetin, quercetin, paniculate biflavonoids, and quercetin, thereby having significant antioxidant and anti-inflammatory effects and being mild and non-irritating. DETAILED DESCRIPTION

[0032] In order to make the technical scheme of the present invention more clearly understood by those skilled in the art, the following embodiments are listed for illustration. It should be pointed out that the following embodiments do not limit the protection scope of the present invention.

[0033] Unless otherwise specified, the raw materials, reagents or devices used in the following examples can be obtained from conventional commercial sources or by existing known methods. Unless otherwise specified, the experiments or test methods are conventional methods in the art.

[0034] Example 1

[0035] A method for preparing a Platycladus orientalis leaf extract comprises the following steps:

[0036] 1) 75 g of urea and 225 g of sorbitol were mixed, and the mixture was heated at 90° C. until the powder was completely dissolved to obtain a low eutectic solvent; 60 g of the low eutectic solvent and 140 g of water were then mixed to obtain an extraction solvent;

[0037] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0038] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0039] Example 2

[0040] A method for preparing a Platycladus orientalis leaf extract comprises the following steps:

[0041] 1) 120 g of urea and 180 g of trehalose were mixed, and the mixture was heated at 90° C. until the powder was completely dissolved to obtain a low eutectic solvent; 60 g of the low eutectic solvent and 140 g of water were then mixed to obtain an extraction solvent;

[0042] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0043] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0044] Example 3

[0045] A method for preparing a Platycladus orientalis leaf extract comprises the following steps:

[0046] 1) After mixing 100 g of betaine and 200 g of sorbitol, heat at 90° C. until the powder is completely dissolved to obtain a low eutectic solvent; then mix 60 g of the low eutectic solvent and 140 g of water to obtain an extraction solvent;

[0047] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0048] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0049] Example 4

[0050] A method for preparing a Platycladus orientalis leaf extract comprises the following steps:

[0051] 1) 180 g of betaine and 120 g of trehalose were mixed, and the mixture was heated at 90° C. until the powder was completely dissolved to obtain a low eutectic solvent; 60 g of the low eutectic solvent and 140 g of water were then mixed to obtain an extraction solvent;

[0052] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0053] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0054] Example 5

[0055] A method for preparing a Platycladus orientalis leaf extract comprises the following steps:

[0056] 1) 75 g of urea and 225 g of sorbitol were mixed, and the mixture was heated at 90° C. until the powder was completely dissolved to obtain a low eutectic solvent; 140 g of the low eutectic solvent and 60 g of water were then mixed to obtain an extraction solvent;

[0057] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0058] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0059] Example 6

[0060] A method for preparing a Platycladus orientalis leaf extract comprises the following steps:

[0061] 1) 120 g of urea and 180 g of trehalose were mixed, and the mixture was heated at 90° C. until the powder was completely dissolved to obtain a low eutectic solvent; 140 g of the low eutectic solvent and 60 g of water were then mixed to obtain an extraction solvent;

[0062] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0063] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0064] Example 7

[0065] A method for preparing a Platycladus orientalis leaf extract comprises the following steps:

[0066] 1) After mixing 100 g of betaine and 200 g of sorbitol, heat at 90° C. until the powder is completely dissolved to obtain a low eutectic solvent; then mix 140 g of the low eutectic solvent and 60 g of water to obtain an extraction solvent;

[0067] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0068] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0069] Example 8

[0070] A method for preparing a Platycladus orientalis leaf extract comprises the following steps:

[0071] 1) 180 g of betaine and 120 g of trehalose were mixed, and the mixture was heated at 90° C. until the powder was completely dissolved to obtain a low eutectic solvent; 140 g of the low eutectic solvent and 60 g of water were then mixed to obtain an extraction solvent;

[0072] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0073] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0074] Comparative Example 1

[0075] A method for preparing a Platycladus orientalis leaf extract comprises the following steps:

[0076] 1) After mixing 10 g of Platycladus orientalis leaves and 100 g of water, the mixture was added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0077] 2) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf water extract.

[0078] Comparative Example 2

[0079] 1) Mix 60 g of urea and 140 g of water to obtain an extraction solvent;

[0080] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0081] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0082] Comparative Example 3

[0083] 1) Mix 60 g of betaine and 140 g of water to obtain an extraction solvent;

[0084] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0085] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0086] Comparative Example 4

[0087] 1) Mix 60 g of sorbitol and 140 g of water to obtain an extraction solvent;

[0088] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0089] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0090] Comparative Example 5

[0091] 1) Mix 60 g of trehalose and 140 g of water to obtain an extraction solvent;

[0092] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0093] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0094] Comparative Example 6

[0095] 1) 10 g of Platycladus orientalis leaves, 7.5 g of urea, 22.5 g of sorbitol, and 70 g of water were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0096] 2) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0097] Comparative Example 7

[0098] 1) After mixing 10 g of Platycladus orientalis leaves, 18 g of betaine, 12 g of trehalose and 70 g of water, the mixture was added into a high-pressure homogenizer and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0099] 2) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0100] Comparative Example 8

[0101] 1) 10 g of Platycladus orientalis leaves and 100 g of 20% ethanol (V / V) were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0102] 2) The Platycladus orientalis leaf homogenate was heated and extracted at 80° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0103] Comparative Example 9

[0104] 1) 10 g of Platycladus orientalis leaves and 100 g of 50% ethanol (V / V) were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0105] 2) The Platycladus orientalis leaf homogenate was heated and extracted at 80° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0106] Comparative Example 10

[0107] 1) 75 g of urea and 225 g of glucose were mixed, and the mixture was heated at 90° C. until the powder was completely dissolved to obtain a low eutectic solvent; 60 g of the low eutectic solvent and 140 g of water were then mixed to obtain an extraction solvent;

[0108] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0109] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0110] Comparative Example 11

[0111] 1) 100 g of citric acid and 200 g of sorbitol were mixed, and the mixture was heated at 90° C. until the powder was completely dissolved to obtain a low eutectic solvent; 140 g of the low eutectic solvent and 60 g of water were then mixed to obtain an extraction solvent;

[0112] 2) 10 g of Platycladus orientalis leaves and 100 g of extraction solvent were mixed, and then added into a high-pressure homogenizer, and homogenized for 15 min at a high-pressure homogenization temperature of 40° C. and a high-pressure homogenization pressure of 80 MPa to obtain a Platycladus orientalis leaf homogenate;

[0113] 3) The Platycladus orientalis leaf homogenate was heated and extracted at 100° C. for 20 min, and filtered to obtain the Platycladus orientalis leaf extract.

[0114] Test Example 1: Determination of total flavonoid content

[0115] This test example tests the total flavonoid content in the Platycladus orientalis leaf extracts of the above Examples 1-8 and the Platycladus orientalis leaf extracts of Comparative Examples 1-11 (hereinafter referred to as "samples"). The specific test method is as follows:

[0116] Adjust the total flavonoids concentration of the sample solution to the measurement range, accurately pipette 2.0 mL of sample into a dry, clean test tube, add 0.2 mL of 5% sodium nitrite solution, shake well and let stand for 5 minutes, then add 0.2 mL of 10% aluminum nitrate solution and shake well, let stand for 6 minutes, add 1.0 mL of 5% sodium hydroxide solution, add 50% ethanol (V / V) to the scale, and shake well to obtain the sample solution.

[0117] The sample solution was rapidly measured for absorbance of each tube at a wavelength of 510 nm. The concentration of total flavonoids was calculated from the absorbance of the sample solution according to the standard curve of rutin solution, and the total flavonoid content in the sample was expressed as rutin.

[0118] Test Example 2: Determination of the content of myricitrin, isoquercetin, quercetin, spicateflavonoids and quercetin

[0119] This test example tests the contents of myricitrin, isoquercitrin, quercitrin, agnathin and quercetin in the Platycladus orientalis leaf extracts of Examples 1-8 and Comparative Examples 1-11 (hereinafter referred to as "samples"). The specific test method is as follows:

[0120] Preparation of test samples: Take 2.0 g of sample and place it in a 10 mL volumetric flask, add chromatographic grade methanol to the mark, shake well, and filter with a 0.45 μm filter membrane to obtain the test sample.

[0121] Chromatographic conditions: Use an Agilent ZORBAX SB-C18 column (4.6 mm × 250 mm, 5 μm); use 0.05% phosphoric acid (A)-acetonitrile (B) as the mobile phase for gradient elution (0 min, 17% B; 20 min, 17% B; 20 min, 17% B; 60 min, 50% B), flow rate 1.0 mL / min; detection wavelength 360 nm; column temperature 30 ° C.

[0122] The test results of Experimental Example 1 and Experimental Example 2 are shown in Table 1.

[0123] Table 1

[0124]

[0125]

[0126] As shown in Table 1, compared with Comparative Examples 1-8, the contents of total flavonoids, myricitrin, isoquercitrin, quercitrin, quercetin and quercetin in the samples of Examples 1-8 are greatly improved. The hydrogen bond acceptor component in the deep eutectic solvent used in the extraction process of the present invention contains nitrogen atoms, and the flavonoid components in the Platycladus orientalis leaves will further form hydrogen bonds and van der Waals forces with the components in the deep eutectic solvent, and more binding forces are introduced into the system. This interaction not only does not destroy the internal structure of the original deep eutectic solvent, but makes the structure of the system more stable, and more effectively and targetedly helps the extraction solvent to efficiently extract the active ingredients. At the same time, the extraction solvent can efficiently penetrate into the Platycladus orientalis leaf tissue, quickly form a concentration gradient, and make the active ingredients more easily dissolved.

[0127] Compared with Comparative Examples 10 and 11, different deep eutectic solvents have different extraction efficiencies for flavonoid components in Platycladus orientalis leaves, indicating that the deep eutectic solvents and Platycladus orientalis leaf components are compatible and specific during the extraction process. In addition, compared with Comparative Example 9, the total flavonoid content and the content of each compound are also improved, indicating that the present invention can avoid the use of organic solvents and the risk of organic solvent residues when efficiently obtaining Platycladus orientalis leaf flavonoid components, and achieve green and efficient extraction of active substances.

[0128] Test Example 3: DPPH free radical scavenging ability

[0129] This test example tests the DPPH free radical scavenging ability of the Platycladus orientalis leaf extracts of Examples 1-8 and Comparative Examples 1-9 (hereinafter referred to as "samples"). The specific test method is as follows:

[0130] Add 2 mL of DPPH test solution with a mass concentration of 45.8 mg / L and 0.5 mL of the sample to be tested to a test tube, add 50% ethanol (V / V) to make the total volume 3 mL, shake well, react in the dark for 30 minutes, and measure the absorbance at a wavelength of 517 nm using a 1 cm cuvette, which is recorded as A1; add 2 mL of anhydrous ethanol and the corresponding volume of the sample to be tested to a test tube, add 50% ethanol (V / V) to make the total volume 3 mL, and the measured absorbance is recorded as A2; add 2 mL of DPPH test solution and 1 mL of 50% ethanol (V / V) to a test tube, and the measured absorbance is recorded as A3.

[0131] The calculation formula is as follows:

[0132] DPPH free radical scavenging rate = (1-(A3-A2) / A1) × 100%.

[0133] IC calculation using R language 50 value.

[0134] Test Example 4: OH radical scavenging ability

[0135] This test example tests the OH radical scavenging ability of the Platycladus orientalis leaf extracts of Examples 1-8 and Comparative Examples 1-9 (hereinafter referred to as "samples"). The specific test method is as follows:

[0136] First add 1.0mL FeSO4 solution (9.0mmol / L), 1mL salicylic acid-ethanol solution (9.0mmol / mL), and 1.0mL sample into the reaction system, then add 1.0mL H2O2 (8.8mmol / mL), and react in a 37℃ water bath for 30min. Use anhydrous ethanol as a blank control, and measure the absorbance at 510nm.

[0137] The calculation formula is as follows:

[0138] OH radical scavenging rate = [1-(A1-A2) / A3] × 100%, where A1 is the absorbance of the extract reaction system; A2 is the absorbance of the sample without H2O2 (with anhydrous ethanol instead of H2O2); A3 is the absorbance of the blank control anhydrous ethanol.

[0139] IC calculation using R language 50 value.

[0140] Test Example 5: Determination of total antioxidant capacity by FRAP method

[0141] This test example tests the total antioxidant capacity of the Platycladus orientalis leaf extracts of Examples 1-8 and Comparative Examples 1-9 (hereinafter referred to as "samples"). The specific test method is as follows:

[0142] Sample determination: Add 100 μL of sample solution to the reaction tube, then add 3 mL of FRAP working solution, mix well, react at 37°C for 15 min, and measure the absorbance at 593 nm.

[0143] Determination of FeSO4 standard curve: According to the determination steps of the sample, the standard curve is drawn with 0.1-1.6mmol / L FeSO4 standard solution instead of the sample. The final total antioxidant activity of the sample is expressed as the equivalent concentration of FeSO4, in mmol / L. With FeSO4 as the standard solution, the corresponding FeSO4 concentration (mmol / L) is obtained on the standard curve according to the absorbance value after the reaction, which is defined as the FRAP value. The larger the FRAP value, the stronger the antioxidant activity.

[0144] The test results of Test Example 3, Test Example 4 and Test Example 5 are shown in Table 2.

[0145] Table 2

[0146]

[0147] As shown in Table 2, compared with Comparative Examples 1-9, the DPPH free radical scavenging rates IC 50 IC value and OH radical scavenging rate 50 The values ​​are all lower, indicating that the free radical scavenging ability is stronger. In addition, in terms of total antioxidant capacity, the Platycladus orientalis leaf extracts of Examples 1-8 have a stronger effect.

[0148] Test Example 6: Evaluation of anti-inflammatory activity

[0149] In this test example, the anti-inflammatory activity of Platycladus orientalis leaf extract was evaluated by the production of nitric oxide (NO), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in mouse macrophage RAW264.7 cells induced by lipopolysaccharide (LPS).

[0150] Mouse macrophage RAW264.7 cells were cultured in DMEM complete medium (DMEM medium + 10% fetal bovine serum + 1% double antibody) in an incubator at 37°C and 5% CO2, and passaged every 2 to 3 days. RAW264.7 cells in the logarithmic growth phase were cultured at 1×10 5 / well was inoculated in a 96-well plate, with 200 μL of DMEM complete medium in each well. After culturing for 24 hours, 1 μg / mL of LPS and 4% of each sample were added, 4% of DMEM complete medium was added to the blank group, and 1 μg / mL of LPS and 4% of DMEM complete medium were added to the model group. The culture was continued for 24 hours, and the supernatant was collected. The production of NO, COX-2, TNF-α and IL-6 was determined according to the instructions of the kit. The test results are shown in Table 3.

[0151] Table 3

[0152]

[0153]

[0154] It can be seen from Table 3 that, compared with Comparative Examples 1-9, the Platycladus orientalis leaf extracts of Examples 1-8 can more effectively reduce the production of NO, COX-2, TNF-α and IL-6, showing a stronger anti-inflammatory effect.

[0155] Test Example 7: Eye Irritation / Corrosion Test on Chicken Embryo Chorioallantoic Membrane

[0156] Select 9-day-old chicken embryos with a mass of 50-60g, mark the position of the air chamber on the eggshell surface, peel off the eggshell part with tweezers to expose the white egg membrane, add 0.9% saline with a pipette to moisten the egg membrane, remove the inner membrane with tweezers, take 0.3mL sample and directly act on the chorioallantoic membrane (CAM), ensuring that the coverage area reaches at least 50%. Immediately observe the CAM reaction until 5min stop, record the observation results and make a judgment, repeat each sample 3 times, and use saline as a negative control. Among them, IS<1 is non-irritating, 1≤IS<5 is mildly irritating, and 5≤IS<10 is moderately irritating. The test results are shown in Table 4.

[0157] Table 4

[0158]

[0159]

[0160] It can be seen from Table 4 that the Platycladus orientalis leaf extracts of Examples 1-8, Comparisons 1-7 and Comparative Example 10 are non-irritating, and the Platycladus orientalis leaf extracts of Comparative Example 8, Comparative Example 9 and Comparative Example 11 are slightly irritating, indicating that the Platycladus orientalis leaf extracts prepared in the embodiments of the present invention are relatively mild raw materials.

[0161] Application Example 1

[0162] A shampoo, the formula of which is shown in Table 5.

[0163] Table 5

[0164] Element Mass percentage (%) Deionized water margin Sodium Laureth Sulfate 14.0 Cocamidopropyl Betaine 8.50 Cocamide Methyl MEA 2.80 Example 8 Platycladus orientalis leaf extract 2.0 PPG-3 Octyl Ether 0.60 Polyquaternium-10 0.40 Sodium Benzoate 0.30 Phenoxyethanol 0.50 Sodium chloride 0.20 Disodium EDTA 0.10 Citric Acid 0.10 Sodium Citrate 0.10

[0165] The shampoo is prepared by a conventional preparation method.

[0166] Application Comparative Example 1

[0167] The only difference from Application Example 1 is that the Platycladus orientalis leaf extract in Example 8 is replaced by the Platycladus orientalis leaf water extract in Comparative Example 1 in equal amounts, and the remaining ingredients, amounts used and preparation methods are the same as those in Application Example 1.

[0168] Test Example 8: Evaluation of the soothing effect of shampoo

[0169] Test samples: shampoos provided in Application Example 1 and Application Comparative Example 1.

[0170] 60 volunteers were randomly divided into 2 groups, aged between 20 and 50 years old, half male and half female; 5g of shampoo of application example 1 and comparative example 1 were used on the 1st, 3rd, 5th and 7th day, respectively, and the subjects were interviewed on the 8th day to evaluate the effects, and the subjects scored according to their feelings. The scoring standard adopts a 10-point system, and the higher the score, the better the feeling of use.

[0171] Improvement of itching (0 means no improvement, 10 means significant improvement); overall feeling of use (0 means very bad, 10 means very good).

[0172] The test results are shown in Table 6 (average scores).

[0173] Table 6

[0174] sample Anti-itching effect Overall use experience Application Example 1 8.6 8.3 Application Comparative Example 1 7.2 7.5

[0175] As shown in Table 6, compared with the comparative example 1, the volunteers had a significant anti-itching effect after using the shampoo containing the Platycladus orientalis leaf extract of the present invention, and the overall feeling of use was greatly improved. It further shows that the Platycladus orientalis leaf extract of the present invention can not only inhibit the production of NO, COX-2, TNF-α and IL-6, but also scavenge free radicals. The anti-itching effect of the shampoo is related to the anti-inflammatory and antioxidant effects of the raw materials.

[0176] Test Example 9: Evaluation of the Oil Control Effect of Shampoo

[0177] Test samples: shampoos provided in Application Example 1 and Application Comparative Example 1.

[0178] Recruit 20 volunteers with strong scalp oil secretion, aged 20-30. If the volunteers are female, avoid the menstrual cycle, have a regular life and work schedule, and try to avoid strenuous exercise, sun exposure, staying up late, etc.

[0179] Do not wash your hair the night before the test. Before the test, the volunteers sit quietly for 30 minutes in a room with a temperature of 20-22℃ and a humidity of 40%-65%, and cannot drink water or beverages during this period. The test site is exposed and the volunteers remain relaxed.

[0180] The test time points are before using the sample, immediately after using the sample, 6 hours, and 8 hours. The test site is the 5cm area from the forehead to the top of the head. Use a skin pen to draw a 1cm×1cm square in the test area. When using the sample, wash half of the head. Divide the head along the hairline in the middle, half of which is the blank control and washed with water. The other half is the sample area, and the test sample is washed with the hair.

[0181] Bend the tape into a ring and make it contact with the scalp. The lipid compounds (oils) in the scalp will change the transparency of the test tape. Then flatten the test tape and put it in the slider of the instrument, and push the slider into the instrument. The transmittance distribution curve on the tape can be tested by the photoelectric tube in the instrument. The peak value of the transmittance curve distribution represents the content of lipid compounds, i.e. oil, in the scalp. During the test, the top of the ring of the tape is exactly the part that contacts the scalp.

[0182] Calculation of the mean difference in scalp oil content: the mean difference in scalp oil content between the blank control group and the sample group at different time periods = the scalp oil content of the blank control group - the scalp oil content of the sample group.

[0183] The test results are shown in Table 7.

[0184] Table 7

[0185]

[0186] It can be seen from Table 7 that the shampoo containing the Platycladus orientalis leaf extract of the present invention can slow down the secretion of scalp oil and achieve a good oil control effect.

[0187] The preferred embodiments of the present invention are specifically described above, but the invention is not limited to the embodiments. Those skilled in the art may make various equivalent modifications or substitutions without violating the spirit of the invention. These equivalent modifications or substitutions are all included in the scope defined by the claims of this application.

Claims

1. A method for preparing a Platycladus orientalis leaf extract, characterized in that: The following steps are involved: The Platycladus orientalis leaves and the extraction solvent are mixed and homogenized under high pressure to obtain a Platycladus orientalis leaf homogenate; The Platycladus orientalis leaf homogenate is subjected to heating extraction and filtering to obtain the Platycladus orientalis leaf extract; The extraction solvent includes a deep eutectic solvent and water; The deep eutectic solvent comprises a hydrogen bond acceptor and a hydrogen bond donor, wherein the hydrogen bond acceptor comprises urea and / or betaine, and the hydrogen bond donor comprises sorbitol and / or trehalose.

2. The preparation method according to claim 1, characterized in that: The usage ratio of the Platycladus orientalis leaves to the extraction solvent is 1g:(0.5-50g).

3. The preparation method according to claim 1, characterized in that: The usage ratio of the low eutectic solvent to the water is 1g:(0.05-20g).

4. The preparation method according to claim 1, characterized in that: The usage ratio of the hydrogen bond acceptor to the hydrogen bond donor is 1g:(0.2-5g).

5. The preparation method according to claim 1, characterized in that: The pressure of the high-pressure homogenization is 20-100 MPa; and / or, the temperature of the high-pressure homogenization is 25-50° C.; and / or, the time of the high-pressure homogenization is 5-40 min.

6. The preparation method according to claim 1, characterized in that: The temperature of the heating extraction is 90-110° C.; and / or the time of the heating extraction is 10-40 min.

7. A Platycladus orientalis leaf extract, characterized in that The extract is prepared by the preparation method according to any one of claims 1 to 6, and the content of total flavonoids in the Platycladus orientalis leaf extract is greater than 20 mg / g.

8. A cosmetic, characterized in that: The invention comprises the Platycladus orientalis leaf extract as claimed in claim 7.

9. The cosmetic according to claim 8, characterized in that: The content of the Platycladus orientalis leaf extract is 0.01wt%-50wt%.

10. The cosmetic according to claim 8, characterized in that: The cosmetic is selected from any one of shampoo, conditioner, hair oil, shower gel, facial cleanser, facial mask, lotion, facial cream or essence.

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