Lycopodium nutans active compound as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of plant extracts, and particularly discloses a lycopodium nutans active compound as well as a preparation method and application thereof, the structure of the compound is as follows: # imgabs0 #, iI is huperzene glycol; iII is 2, 4-di-tert-butyl phenol; iV is 5-hydroxymethylfurfural; v is 5-ethyoxyl methyl furfural; vI is ethyl palmitate; vII is ethyl linoleate; and VIII is 1-eicosene. According to the lycopodium nudum active compound and the preparation method and application thereof, a new compound is found, the tyrosinase activity of the compound is equivalent to that of a positive control, and the technical problems that an existing whitening substance has the risks of allergy, carcinogenesis and the like and is prone to discoloration due to environmental influence, and the whitening effect is affected are solved; the lycopodium nutans active compound with a good whitening effect is extracted by a simple extraction method.

Description

Technical Field

[0001] The invention belongs to the technical field of plant extract technology, and specifically relates to an active compound of Lycopodiella nutans and a preparation method and application thereof. Background Art

[0002] Tyrosinase plays many different roles in humans, insects, plants and microorganisms. In humans, tyrosinase is involved in the formation of melanin. The melanin produced by tyrosinase determines the color of the skin, hair and iris in the human body, and can also protect the skin from harmful effects such as ultraviolet (UV) radiation, oxidative stress and DNA damage. However, excessive accumulation of melanin caused by abnormal expression of tyrosinase can cause pigmentation diseases such as melanoma, chloasma and freckles. As the research on tyrosinase continues to deepen, more and more tyrosinase inhibitors are being developed and studied, and natural compounds are an important source of tyrosinase inhibitors. In the current cosmetics market, most whitening products are based on tyrosinase activity inhibitors as the main ingredients, and most whitening substances, such as hydroquinone and vitamin C, achieve whitening effects by inhibiting tyrosinase or antioxidant activity. Although hydroquinone has a significant whitening effect, it is banned due to the risk of carcinogenesis. Vitamin C is easily affected by factors such as light, heat, humidity and pH value during the preparation process and changes color. There is currently no report on the use of Lycopodiella nutans in whitening products on the market.

[0003] Lycopodiella danglingensis is a plant of the Lycopodiella genus of the Lycopodiaceae family. It is a medium to large native plant and is produced in the forest, forest edge, shade under the bushes or on rocks at an altitude of 100-1800 meters in Zhejiang, Jiangxi, Fujian, Taiwan, Hunan, Guangdong, Hong Kong, Guangxi, Hainan, Sichuan, Chongqing, Guizhou, Yunnan, etc. According to the "Chinese Materia Medica", Lycopodiella danglingensis is used as medicine with the whole plant. It tastes bitter and is flat in nature. It enters the liver, spleen, and kidney meridians and has the effects of dispelling wind and dampness, relaxing muscles and activating blood circulation, and relieving cough and diuresis. Modern research shows that Lycopodiella danglingensis mainly contains chemical components such as alkaloids, triterpenes, and flavonoids, which have anti-inflammatory, cytotoxic, acetylcholinesterase, α-glucosidase, and xanthine oxidase effects. Therefore, Lycopodiella danglingensis has broad application prospects as a complementary alternative medicine. However, up to now, no research reports on the tyrosinase activity of Lycopodiella danglingensis have been found.

[0004] Therefore, the art needs to develop an active compound of Lycopodiella cuneata and its preparation method and application, which can effectively solve the technical problems that existing whitening substances have risks such as allergies and carcinogenicity, and are easily discolored by environmental influences, thus affecting the whitening effect. Summary of the invention

[0005] The purpose of the present invention is to provide an active compound of Lycopodiella cuneata and a preparation method and application thereof. A new compound is discovered whose tyrosinase activity is equivalent to that of a positive control, which solves the technical problems that existing whitening substances have risks such as allergies and carcinogenicity and are easily discolored by environmental influences, thereby affecting the whitening effect; the active compound of Lycopodiella cuneata with good whitening effect is extracted by a simple extraction method.

[0006] To achieve the above object, the present invention provides a Lycopodiella foetida active compound, the structure of which comprises:

[0007]

[0008] Among them, I is furinol; II is melaleuca glycol; III is 2,4-di-tert-butylphenol; IV is 5-hydroxymethylfurfural; V is 5-ethoxymethylfurfural; VI is ethyl palmitate; VII is ethyl linoleate; and VIII is 1-eicosene.

[0009] A method for preparing an active compound of Lycopodiella foetida comprises the following steps:

[0010] Step S1, naturally air-drying and crushing the Lycopodiella ciliata sample to obtain powder;

[0011] Step S2, soaking the powder obtained in step S1 with anhydrous ethanol, and concentrating under reduced pressure to obtain an ethanol extract;

[0012] Step S3, dispersing the ethanol extract obtained in step S2, and extracting with petroleum ether and chloroform in sequence to obtain petroleum ether extract and chloroform extract;

[0013] Step S4, separating the petroleum ether extract or chloroform extract obtained in step S3 on a 300-400 mesh silica gel chromatography column, eluting with petroleum ether-ethyl acetate mobile phase to obtain multiple components; further separating and purifying, and obtaining the active compounds of Lycopodiella foetida through an elution procedure.

[0014] Preferably, in step S2, the powder obtained in step S1 is soaked in anhydrous ethanol for 3 times, each time for 7 days; the volume ratio of anhydrous ethanol to powder is 1:1.

[0015] Preferably, in step S2, the temperature of reduced pressure concentration is 37°C, the vacuum degree is 0.1 MPa, the rotation speed is 70 rpm / min, and the condensed water temperature is 5°C.

[0016] Preferably, in step S4, the silica gel chromatography column is one or both of a 300-400 mesh silica gel chromatography column or a Sephadex LH-20 gel column chromatography column.

[0017] Preferably, in step S4, the mobile phase in the elution procedure is one or more of petroleum ether-ethyl acetate, dichloromethane-methanol, and petroleum ether-dichloromethane-methanol.

[0018] Application of an active compound of Lycopodiella nutans in the preparation of a tyrosinase inhibitor.

[0019] Application of an active compound of Lycopodiella nutans in the preparation of skin care products.

[0020] The present invention adopts the above-mentioned Lycopodiella cuneata active compound and its preparation method and application, and the beneficial effects are as follows:

[0021] (1) The active compounds in the Lycopodiella truncatula of the present invention are used to prepare whitening skin care products, which have a better inhibitory effect on tyrosinase activity;

[0022] (2) The present invention has discovered a new compound whose tyrosinase activity is equivalent to that of the positive control, thus solving the technical problems of the existing whitening substances such as the risks of allergy and carcinogenesis and the easy discoloration due to environmental influences, thereby affecting the whitening effect;

[0023] (3) The present invention provides a simple extraction method, which comprises drying the whole herb of Lycopodiella ciliata, crushing it, soaking it in anhydrous ethanol, and concentrating it under reduced pressure to obtain an extract; after dispersing the ethanol extract, extracting it with petroleum ether and chloroform in sequence to obtain petroleum ether extract and chloroform extract; eluting the extract and then extracting the active compounds of Lycopodiella ciliata with good whitening effect.

[0024] The technical solution of the present invention is further described in detail below through the accompanying drawings and embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 The present invention provides a preparation flow chart of an active compound of Lycopodiella foetida and its preparation method and application example. DETAILED DESCRIPTION

[0026] The technical solution of the present invention is further described below through the accompanying drawings and embodiments.

[0027] Unless otherwise defined, technical or scientific terms used in the present invention shall have the common meanings understood by one having ordinary skills in the field to which the present invention belongs.

[0028] Embodiment 1

[0029] like Figure 1 As shown, a method for preparing an active compound of Lycopodiella foetida comprises the following steps:

[0030] Step S1, naturally air-dry and crush 450 g of Lycopodiella cuneata sample to obtain powder.

[0031] Step S2, soaking the powder obtained in step S1 with anhydrous ethanol, the volume ratio of anhydrous ethanol to powder is 1:1. Soaking for 3 times, each time for 7 days, the temperature is 37°C, the vacuum degree is 0.1MPa, the rotation speed is 70rpm / min, the condensed water temperature is 5°C, and the reduced pressure concentration is obtained to obtain 415g of ethanol extract.

[0032] Step S3, after dispersing the ethanol extract obtained in step S2, extracting with petroleum ether and chloroform in sequence to obtain petroleum ether extract and chloroform extract.

[0033] Step S4, separating the chloroform extract obtained in step S3 on a 300-400 mesh silica gel chromatography column, and eluting with a petroleum ether-ethyl acetate mobile phase at a gradient of 20:1, 10:1, 8:2, 7:3, 1:1, and 0:1 to obtain 6 components (Fr.C1-Fr.C6).

[0034] Component Fr.C5 was separated on a 300-400 mesh silica gel column chromatography column and eluted with a petroleum ether-ethyl acetate mobile phase in a gradient of 8:2, 7:3, 6:4, and 0:1 to obtain 20 subcomponents (Fr.C5A-Fr.C5T).

[0035] The subcomponent Fr.C5S was separated on a 300-400 mesh silica gel column, eluted with petroleum ether-ethyl acetate mobile phase at a ratio of 1:1, and then eluted with Sephadex LH-20 gel column chromatography, using petroleum ether-dichloromethane-methanol mobile phase at a ratio of 2:1:1. Multiple rounds of repeated elutions gave the active compound I of Lycopodiella foetida, whose structural formula is:

[0036]

[0037] Embodiment 2

[0038] A method for preparing an active compound of Lycopodiella foetida comprises the following steps:

[0039] Step S1, naturally air-dry and crush 450 g of Lycopodiella cuneata sample to obtain powder.

[0040] Step S2, soaking the powder obtained in step S1 with anhydrous ethanol, the volume ratio of anhydrous ethanol to powder is 1:1. Soaking for 3 times, each time for 7 days, the temperature is 37°C, the vacuum degree is 0.1MPa, the rotation speed is 70rpm / min, the condensed water temperature is 5°C, and the reduced pressure concentration is obtained to obtain 415g of ethanol extract.

[0041] Step S3, after dispersing the ethanol extract obtained in step S2, extracting with petroleum ether and chloroform in sequence to obtain petroleum ether extract and chloroform extract.

[0042] Step S4, separating the chloroform extract obtained in step S3 on a 300-400 mesh silica gel chromatography column, and eluting with a petroleum ether-ethyl acetate mobile phase at a gradient of 20:1, 10:1, 8:2, 7:3, 1:1, and 0:1 to obtain 6 components (Fr.C1-Fr.C6).

[0043] Component Fr.C5 was separated on a 300-400 mesh silica gel column chromatography column and eluted with a petroleum ether-ethyl acetate mobile phase in a gradient of 8:2, 7:3, 6:4, and 0:1 to obtain 20 subcomponents (Fr.C5A-Fr.C5T).

[0044] The subcomponent Fr.C5L was separated on a 300-400 mesh silica gel column and eluted with a dichloromethane-methanol mobile phase at a ratio of 100:1 to obtain the active compound II of Lycopodiella foetida, whose structural formula is:

[0045]

[0046] Embodiment 3

[0047] A method for preparing an active compound of Lycopodiella foetida comprises the following steps:

[0048] Step S1, naturally air-dry and crush 450 g of Lycopodiella cuneata sample to obtain powder.

[0049] Step S2, soaking the powder obtained in step S1 with anhydrous ethanol, the volume ratio of anhydrous ethanol to powder is 1:1. Soaking for 3 times, each time for 7 days, the temperature is 37°C, the vacuum degree is 0.1MPa, the rotation speed is 70rpm / min, the condensed water temperature is 5°C, and the reduced pressure concentration is obtained to obtain 415g of ethanol extract.

[0050] Step S3, after dispersing the ethanol extract obtained in step S2, extracting with petroleum ether and chloroform in sequence to obtain petroleum ether extract and chloroform extract.

[0051] Step S4, separating the chloroform extract obtained in step S3 on a 300-400 mesh silica gel chromatography column, and eluting with a petroleum ether-ethyl acetate mobile phase at a gradient of 20:1, 10:1, 8:2, 7:3, 1:1, and 0:1 to obtain 6 components (Fr.C1-Fr.C6).

[0052] Component Fr.C4 was separated on a 300-400 mesh silica gel column and eluted with a petroleum ether-ethyl acetate mobile phase at a ratio of 6:1 to obtain three subcomponents (Fr.C4A-Fr.C4C).

[0053] Subfraction Fr.C4A was chromatographed on a Sephadex LH-20 gel column using a petroleum ether-dichloromethane-methanol mobile phase at a ratio of 2:1:1, and then chromatographed on a 300-400 mesh silica gel column using a petroleum ether-dichloromethane mobile phase at a ratio of 5:1 to obtain the active compound III of Lycopodiella foetida, whose structural formula is:

[0054]

[0055] Embodiment 4

[0056] A method for preparing an active compound of Lycopodiella foetida comprises the following steps:

[0057] Step S1, naturally air-dry and crush 450 g of Lycopodiella cuneata sample to obtain powder.

[0058] Step S2, soaking the powder obtained in step S1 with anhydrous ethanol, the volume ratio of anhydrous ethanol to powder is 1:1. Soaking for 3 times, each time for 7 days, the temperature is 37°C, the vacuum degree is 0.1MPa, the rotation speed is 70rpm / min, the condensed water temperature is 5°C, and the reduced pressure concentration is obtained to obtain 415g of ethanol extract.

[0059] Step S3, after dispersing the ethanol extract obtained in step S2, extracting with petroleum ether and chloroform in sequence to obtain petroleum ether extract and chloroform extract.

[0060] Step S4, separating the chloroform extract obtained in step S3 on a 300-400 mesh silica gel chromatography column, and eluting with a petroleum ether-ethyl acetate mobile phase at a gradient of 20:1, 10:1, 8:2, 7:3, 1:1, and 0:1 to obtain 6 components (Fr.C1-Fr.C6).

[0061] Component Fr.C6 was separated on a 300-400 mesh silica gel column chromatography column and eluted with a dichloromethane-methanol mobile phase at 200:1, 40:1, 10:1, and 1:1 to obtain 7 subcomponents (Fr.C6A-Fr.C6G).

[0062] Subfraction Fr.C6G was chromatographed on a Sephadex LH-20 gel column using a petroleum ether-dichloromethane-methanol mobile phase at a ratio of 2:1:1, and then chromatographed on a 300-400 mesh silica gel column using a dichloromethane-methanol mobile phase at a ratio of 40:1 to obtain the active compound IV of Lycopodiella foetida, the structural formula of which is:

[0063]

[0064] Embodiment 5

[0065] A method for preparing an active compound of Lycopodiella foetida comprises the following steps:

[0066] Step S1, naturally air-dry and crush 450 g of Lycopodiella cuneata sample to obtain powder.

[0067] Step S2, soaking the powder obtained in step S1 with anhydrous ethanol, the volume ratio of anhydrous ethanol to powder is 1:1. Soaking for 3 times, each time for 7 days, the temperature is 37°C, the vacuum degree is 0.1MPa, the rotation speed is 70rpm / min, the condensed water temperature is 5°C, and the reduced pressure concentration is obtained to obtain 415g of ethanol extract.

[0068] Step S3, after dispersing the ethanol extract obtained in step S2, extracting with petroleum ether and chloroform in sequence to obtain petroleum ether extract and chloroform extract.

[0069] Step S4, separating the petroleum ether extract obtained in step S3 on a 300-400 mesh silica gel chromatography column, using petroleum ether-ethyl acetate mobile phase with a gradient elution ratio of 0:1, 20:1, 10:1, 8:2, and 1:1 to obtain 7 components (Fr.P1-Fr.P7).

[0070] Subcomponent Fr.P4 was separated on a 300-400 mesh silica gel column and eluted with a dichloromethane-methanol mobile phase at a ratio of 100:1 to obtain the active compound V of Lycopodiella foetida, the structural formula of which is:

[0071]

[0072] Embodiment 6

[0073] A method for preparing an active compound of Lycopodiella foetida comprises the following steps:

[0074] Step S1, naturally air-dry and crush 450 g of Lycopodiella cuneata sample to obtain powder.

[0075] Step S2, soaking the powder obtained in step S1 with anhydrous ethanol, the volume ratio of anhydrous ethanol to powder is 1:1. Soaking for 3 times, each time for 7 days, the temperature is 37°C, the vacuum degree is 0.1MPa, the rotation speed is 70rpm / min, the condensed water temperature is 5°C, and the reduced pressure concentration is obtained to obtain 415g of ethanol extract.

[0076] Step S3, after dispersing the ethanol extract obtained in step S2, extracting with petroleum ether and chloroform in sequence to obtain petroleum ether extract and chloroform extract.

[0077] Step S4, subjecting the chloroform extract obtained in step S3 to 300-400 mesh silica gel column chromatography, using petroleum ether-ethyl acetate mobile phase with a gradient elution ratio of 20:1, 10:1, 8:2, 7:3, 1:1, and 0:1 to obtain 6 components (Fr.C1-Fr.C6).

[0078] Component Fr.C2 was separated on a 300-400 mesh silica gel column, and eluted with petroleum ether-ethyl acetate mobile phase at 40:1 and 20:1, and then eluted with Sephadex LH-20 gel column chromatography, and eluted with petroleum ether-dichloromethane-methanol mobile phase at 2:1:1 to obtain the active compound VI of Lycopodiella foetida, whose structural formula is:

[0079]

[0080] Embodiment 7

[0081] A method for preparing an active compound of Lycopodiella foetida comprises the following steps:

[0082] Step S1, naturally air-dry and crush 450 g of Lycopodiella cuneata sample to obtain powder.

[0083] Step S2, soaking the powder obtained in step S1 with anhydrous ethanol, the volume ratio of anhydrous ethanol to powder is 1:1. Soaking for 3 times, each time for 7 days, the temperature is 37°C, the vacuum degree is 0.1MPa, the rotation speed is 70rpm / min, the condensed water temperature is 5°C, and the reduced pressure concentration is obtained to obtain 415g of ethanol extract.

[0084] Step S3, after dispersing the ethanol extract obtained in step S2, extracting with petroleum ether and chloroform in sequence to obtain petroleum ether extract and chloroform extract.

[0085] Step S4, subjecting the chloroform extract obtained in step S3 to 300-400 mesh silica gel column chromatography, using petroleum ether-ethyl acetate mobile phase with a gradient elution ratio of 20:1, 10:1, 8:2, 7:3, 1:1, and 0:1 to obtain 6 components (Fr.C1-Fr.C6).

[0086] Component Fr.C2 was separated on a 300-400 mesh silica gel column, and eluted with petroleum ether-ethyl acetate mobile phase at 40:1 and 20:1, and then eluted with Sephadex LH-20 gel column chromatography, and eluted with petroleum ether-dichloromethane-methanol mobile phase at 2:1:1 to obtain the active compound VII of Lycopodiella foetida, whose structural formula is:

[0087]

[0088] Embodiment 8

[0089] A method for preparing an active compound of Lycopodiella foetida comprises the following steps:

[0090] Step S1, naturally air-dry and crush 450 g of Lycopodiella cuneata sample to obtain powder.

[0091] Step S2, soaking the powder obtained in step S1 with anhydrous ethanol, the volume ratio of anhydrous ethanol to powder is 1:1. Soaking for 3 times, each time for 7 days, the temperature is 37°C, the vacuum degree is 0.1MPa, the rotation speed is 70rpm / min, the condensed water temperature is 5°C, and the reduced pressure concentration is obtained to obtain 415g of ethanol extract.

[0092] Step S3, after dispersing the ethanol extract obtained in step S2, extracting with petroleum ether and chloroform in sequence to obtain petroleum ether extract and chloroform extract.

[0093] Step S4, subjecting the chloroform extract obtained in step S3 to 300-400 mesh silica gel column chromatography, using petroleum ether-ethyl acetate mobile phase with a gradient elution ratio of 20:1, 10:1, 8:2, 7:3, 1:1, and 0:1 to obtain 6 components (Fr.C1-Fr.C6).

[0094] The component Fr.C4 was separated on a 300-400 mesh silica gel column and eluted with a petroleum ether-ethyl acetate mobile phase at a ratio of 6:1 to obtain three subcomponents Fr.C4A-Fr.C4C.

[0095] Subfraction Fr.C4A was chromatographed on a 300-400 mesh silica gel column, and eluted with petroleum ether-ethyl acetate mobile phase at 50:1, 20:1, and 7:1 to obtain the active compound VIII of Lycopodiella foetida, whose structural formula is:

[0096]

[0097] The whitening effects of the Lycopodiella active compounds I-VIII prepared in the examples were verified by experiments:

[0098] (1) Preparation of L-DOPA solution: Take 100.0 mg of dry L-DOPA powder, add pH 6.8 phosphate buffer to dissolve it, transfer to a 100 mL brown volumetric flask, and dilute to the mark to obtain a 1.0 mg / mL L-DOPA solution.

[0099] (2) Preparation of tyrosinase solution: Take 2 mg of tyrosinase powder and dissolve it in phosphate buffer with a pH of 6.8. Transfer it to a 100 mL brown volumetric flask and dilute to the mark to obtain 100 U / mL.

[0100] (3) Preparation of reference solution: Weigh 1.0 mg of vitamin C powder, add an appropriate amount of anhydrous ethanol to dissolve, transfer to a 10 mL brown volumetric flask, dilute to the mark to obtain a 1.0 mg / mL vitamin C stock solution, and store at room temperature for later use.

[0101] (4) Preparation of the sample solution to be tested: Prepare a 1.0 mg / mL solution for later use.

[0102] (5) The test sample solution and tyrosinase activity inhibition experimental conditions are shown in Table 1.

[0103] Table 1 Tyrosinase activity inhibition test reaction system

[0104] Reaction system PBS solution Sample solution Tyrosinase Solution L-Tyrosine Solution <![CDATA[A1]]> 50μL 25μL 25μL 100μL <![CDATA[A2]]> 75μL 25μL - 100μL <![CDATA[A3]]> 50μL 25μL 25μL 100μL <![CDATA[A4]]> 75μL 25μL - 100μL

[0105] Use a 96-well plate, add 25 μL of sample solution and 25 μL of tyrosinase solution, then add 100 μL of L-DOPA solution to initiate the reaction, place in an ELISA reader and incubate at 37°C for 30 minutes, scan and detect at a wavelength of 475 nm, and record the data.

[0106] The absorbance of different extract samples was calculated, with vitamin C as the positive control. Three wells were set up in each group, and the average value was taken. The inhibition rate was calculated as follows:

[0107] Inhibition rate (%) = [1-(A1-A2) / (A3-A0)] × 100%,

[0108] In the formula: A1 is the sample group, A2 is the sample group control group, A3 is the blank group, and A0 is the blank control group.

[0109] (6) Comparison of the chemical components of the active compounds of Lycopodiella foetida prepared in the example with the inhibitory capacity of vitamin C on tyrosinase activity IC 50 The values ​​are shown in Table 2.

[0110] Table 2 Inhibition ability to tyrosinase

[0111] Group <![CDATA[IC 50 (mM)]]> Embodiment 1 0.988 Embodiment 2 1.350 Embodiment 3 3.690 Embodiment 4 7.959 Embodiment 5 6.469 Embodiment 6 3.216 Embodiment 7 3.324 Embodiment 8 2.437 Control group 0.984

[0112] As shown in Table 2, the active compounds I-VIII prepared in the examples all have inhibitory effects on tyrosinase. Among them, the active compound I provided in Example 1 exhibits an inhibitory activity comparable to that of the positive control, IC 50 The value is 0.988mM. The active compounds II-VIII prepared in Example 2-Example 8 have good inhibitory activity, IC 50 The values ​​are 1.350mM, 3.690mM, 7.959mM, 6.469mM, 3.216mM, 3.324mM, and 2.437mM respectively.

[0113] The results show that the active compounds of Lycopodiella truncatula prepared in the examples have high potential in the development of whitening products.

[0114] Therefore, the present invention adopts the above-mentioned active compound of Lycopodiella cuneata and its preparation method and application, and discovers a new compound whose tyrosinase activity is equivalent to that of the positive control, which solves the technical problems that existing whitening substances have risks such as allergies and carcinogenicity and are easily discolored by environmental influences, thereby affecting the whitening effect; the active compound of Lycopodiella cuneata with good whitening effect is extracted by a simple extraction method.

[0115] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that they can still modify or replace the technical solution of the present invention with equivalents, and these modifications or equivalent replacements cannot cause the modified technical solution to deviate from the spirit and scope of the technical solution of the present invention.

Claims

1. A Lycopodiella cuneata active compound, characterized in that: The structure of the compound includes: Among them, I is furinol; II is melaleuca glycol; III is 2,4-di-tert-butylphenol; IV is 5-hydroxymethylfurfural; V is 5-ethoxymethylfurfural; VI is ethyl palmitate; VII is ethyl linoleate; and VIII is 1-eicosene.

2. A method for preparing the active compound of Lycopodiella foetida according to claim 1, characterized in that: The following steps are involved: Step S1, naturally air-drying and crushing the Lycopodiella ciliata sample to obtain powder; Step S2, soaking the powder obtained in step S1 with anhydrous ethanol, and concentrating under reduced pressure to obtain an ethanol extract; Step S3, dispersing the ethanol extract obtained in step S2, and extracting with petroleum ether and chloroform in sequence to obtain petroleum ether extract and chloroform extract; Step S4, separating the petroleum ether extract or chloroform extract obtained in step S3 on a 300-400 mesh silica gel chromatography column, eluting with petroleum ether-ethyl acetate mobile phase to obtain multiple components; further separating and purifying, and obtaining the active compounds of Lycopodiella foetida through an elution procedure.

3. The method for preparing a Lycopodiella cuneata active compound according to claim 2, characterized in that: In step S2, the powder obtained in step S1 is soaked in anhydrous ethanol for 3 times, each time for 7 days; the volume ratio of anhydrous ethanol to powder is 1:

1.

4. The method for preparing a Lycopodiella cuneata active compound according to claim 3, characterized in that: In step S2, the temperature of reduced pressure concentration is 37°C, the vacuum degree is 0.1 MPa, the rotation speed is 70 rpm / min, and the condensed water temperature is 5°C.

5. The method for preparing a Lycopodiella cuneata active compound according to claim 1, characterized in that: In step S4, the silica gel chromatography column is one or both of a 300-400 mesh silica gel chromatography column or a Sephadex LH-20 gel column chromatography column.

6. The method for preparing a Lycopodiella cuneata active compound according to claim 5, characterized in that: In step S4, the mobile phase in the elution procedure is one or more of petroleum ether-ethyl acetate, dichloromethane-methanol, and petroleum ether-dichloromethane-methanol.

7. Use of the active compound of Lycopodiella tataricola as claimed in claim 1 in the preparation of a tyrosinase inhibitor.

8. Use of the active compound of Lycopodiella tataricola as claimed in claim 1 in the preparation of skin care products.

Images