Cinnamomum camphora antioxidant active component as well as preparation method and application thereof

Abstract

The invention provides a preparation method of antioxidant active components in Cinnamomum camphora, which comprises the following steps: taking crushed Cinnamomum camphora, adding an ethanol aqueous solution, carrying out heating extraction at 35-70 DEG C, carrying out solid-liquid separation after extraction is ended, concentrating an extracting solution to obtain a Cinnamomum camphora extract, uniformly suspending the Cinnamomum camphora extract in water, sequentially extracting with petroleum ether, ethyl acetate and n-butyl alcohol, and concentrating to obtain the antioxidant active components in the Cinnamomum camphora. Respectively obtaining a petroleum ether layer extract, an ethyl acetate layer extract, an n-butyl alcohol layer extract and an n-butyl alcohol extracted water layer extract; the Cinnamomum camphora antioxidant active component is at least one of an ethyl acetate layer extract, an n-butyl alcohol layer extract and a water layer extract extracted by n-butyl alcohol. According to the invention, the antioxidant active component is prepared from the Cinnamomum camphora for the first time, the cost is low, the operation is simple, the yield is high, the consumed time is short, and further verification by measuring the in-vitro antioxidant capacity shows that the Cinnamomum camphora extract has good antioxidant capacity. The method can provide a theoretical basis for screening antioxidant active components in Cinnamomum camphora.

Description

Technical Field

[0001] The invention belongs to the technical field of antioxidant active components, and particularly relates to a tea fruit camphor antioxidant active component and a preparation method and application thereof. Background Art

[0002] Tea fruit camphor ( Cinnamomum chago BSSun et HLZhao) was published in 1991 (Reference: Sun Bixing; Zhao Huilan. A new species of the genus Cinnamomum from Yunnan [J]. Journal of Yunnan University (Natural Science Edition), 1991, (01): 93-94). It is currently a Class II national key protected wild plant. Due to its rarity and narrow distribution range, it is included in the "Yunnan Province Extremely Small Population Wild Plant Protection List (2021 Edition)" and the IUCN threat level is "Endangered" (EN) (Reference: Zhang X, Zhou XL, Liu YH, et al. Investigating the status of Cinnamomum chago (Lauraceae), a plant species with an extremely small population endemic toYunnan, China. Oryx, 2020, 54(4):470-473. Zhang X, Yang L, Liu YH, et al.Genetic diversity, genetic structure, and demographic history of Cinnamomum chago , a plant species with extremely small populations in China. GlobalEcology and Conservation, 2021, 31: e01808.). In the Flora of China, the genus Cinnamomum in China is divided into two groups: Sect. Camphora and Sect. Cinnamomum.

[0003] The fruit of Cinnamomum camphora contains crude fat, protein, trace elements and health-care ingredients, among which crude fat has the highest content, accounting for 62.4%. There are rich types of fatty acids, with the highest content of lauric acid and capric acid, accounting for 68.92-69.91% and 22.54-23.86% of all fatty acids, respectively. Both are medium-chain saturated fatty acids (reference: Hou Min; Li Junnan; Li Shufang; Yang Jianhua; Xiong Xinwu; Wang Gaosheng. Analysis of nutritional components and nutritional evaluation of Cinnamomum camphora fruit. Western Forestry Science, 2019, 48(06): 80-85.). After entering the human body, medium-chain fatty acids can be directly metabolized in the liver, quickly providing energy for the human body without causing fat accumulation. At the same time, they can inhibit fat production to achieve lipolysis and regulate the structure of intestinal flora (reference: Wang Yuanli; Luan Xia; Yang Jianzhou; Wei Zheng. Current status of development and application of medium-chain fatty acids and their glycerides. China Oils and Fats, 1-13[2023-11-25]). Coconut oil, which is used in the ketogenic diet that has become popular among weight loss people recently, also contains a high proportion of lauric acid and capric acid. Therefore, as people pursue health management, camphor has a good potential for weight loss food development. Camphor kernels contain 7 common minerals, especially rich in potassium ions, which are 1.6 times that of walnuts. At the same time, the sodium ion content is low, and potassium ions are 939 times that of sodium ions. It is a high-potassium and low-sodium food. Potassium ions can fight cancer and prevent cancer, and "high potassium and low sodium" helps maintain the water and salt balance in the human body. Therefore, camphor kernels can be developed as cancer prevention foods and foods for hypertensive patients (References: Hou Min; Li Junnan; Li Shufang; Yang Jianhua; Xiong Xinwu; Wang Gaosheng. Analysis of nutritional components and nutritional evaluation of camphor fruit. Western Forestry Science, 2019, 48 (06): 80-85.). The fruit of Cinnamomum camphora is rich in polyphenols, flavonoids, tannins, total saponins, polysaccharides and crude fiber, but its antioxidant capacity has not been systematically reported. Therefore, the preparation of the antioxidant active ingredients of Cinnamomum camphora and the verification method of the antioxidant activity are studied, which can provide a theoretical basis for the further screening of the main antioxidant active ingredients in Cinnamomum camphora and the development of food or medicine based on Cinnamomum camphora. Based on this, this application was developed. Summary of the invention

[0004] The purpose of the present invention is to overcome the defects of the prior art and provide an antioxidant active component of Cinnamomum camphora. The present invention selects Cinnamomum camphora as the research object, explores the antioxidant effects of different extraction layers of fresh Cinnamomum camphora seeds, and measures the DPPH free radical scavenging ability, ABTS free radical scavenging ability and FRAP reducing ability. The experimental results show that different extraction layers of Cinnamomum camphora seeds have certain antioxidant effects, among which the antioxidant effect of the n-butanol layer is the most significant.

[0005] The invention also provides a preparation method and application of the antioxidant active component of the tea fruit camphor tree.

[0006] To achieve the above object, the present invention adopts the following technical solution: A method for preparing an antioxidant active component of camphor tree, comprising the following steps: 1) Pre-treatment of tea fruit camphor: After peeling off the seed coat and seed shell of fresh Cinnamomum camphora, dry it in the air (usually you can choose to dry it in the sun or dry it naturally for 4-24 hours), crush it and set aside; 2) Preparation of Cinnamomum camphora crude extract: The crushed camphor tree is added with ethanol aqueous solution and heated at 35-70°C for extraction. After the extraction, solid-liquid separation is completed, and the extract is concentrated to obtain camphor tree extract. The camphor tree extract is uniformly suspended in water, and extracted with petroleum ether, ethyl acetate, and n-butanol in sequence to obtain a petroleum ether layer extract, an ethyl acetate layer extract, an n-butanol layer extract, and a water layer extract after n-butanol extraction, respectively; the petroleum ether is mainly used for degreasing; The antioxidant active component of Cinnamomum camphora is at least one of an ethyl acetate layer extract, an n-butanol layer extract and an aqueous layer extract after n-butanol extraction (hereinafter uniformly referred to as "aqueous layer extract").

[0007] Specifically, in step 1), the product can be crushed by a crusher, passed through a 40-100 mesh sieve, and put into a ziplock bag for later use.

[0008] Specifically, in step 2), a 60-85% volume concentration of ethanol aqueous solution is added at a material-liquid ratio of 1 g: 10-30 mL for water bath heating-assisted extraction. The heating extraction time is 2-4 hours, and the extraction is performed 1-4 times.

[0009] Furthermore, in step 2), a vacuum rotary evaporator is used for concentration, and the concentration temperature is 40-60°C.

[0010] Furthermore, in step 2), the Cinnamomum camphora extract is evenly suspended in water at a volume ratio of 1:1-3.

[0011] The invention provides an antioxidant active component of Cinnamomum camphora prepared by the preparation method.

[0012] The present invention provides application of the antioxidant active components of Cinnamomum camphora in preparing antioxidant drugs.

[0013] The present invention provides application of the antioxidant active components of Cinnamomum camphora in preparing a drug for treating or preventing oxidative stress.

[0014] The present invention selects camphor tree as the research object, explores the antioxidant effects of different extraction layers of fresh camphor tree seeds, and determines the DPPH free radical scavenging ability, ABTS free radical scavenging ability and FRAP reducing ability. The experimental results show that different extraction layers of camphor tree seeds have certain antioxidant effects, among which the antioxidant effect of the n-butanol layer extract is the most significant, so the n-butanol layer extract is subjected to SOD detection, CAT detection, GSH detection, MDA detection and apoptosis detection, and it is found that the n-butanol layer has a good antioxidant effect. The purpose of the present invention is to prepare antioxidant active ingredients from camphor tree, detect the content of two antioxidant active ingredients, and explore the antioxidant effects of different extraction layers of camphor tree. In view of the fact that camphor tree is an edible species of extremely small population in the ethnic minority areas of Dali Prefecture, the present invention can provide a theoretical basis for the screening of antioxidant active ingredients in camphor tree, and provide a reference for the development of camphor tree products in related food and cosmetics fields in combination with the verification method of antioxidant activity in camphor tree.

[0015] Compared with the prior art, the present invention has the following advantages and beneficial effects: This paper studies the extraction and antioxidant verification methods of antioxidant active substances in Cinnamomum camphora for the first time, and aims to determine the in vitro antioxidant capacity, combined with water bath heating extraction, and further effectively verify the antioxidant effect. SPSS analysis was used to obtain the IC 50 That is, the sample concentration when scavenging free radicals by 50% was 0.285 ± 0.006 mg / mL and 0.029 ± 0.008 mg / mL for the DPPH free radical scavenging ability of the ethyl acetate layer extract and 0.208 ± 0.002 mg / mL and 0.008 ± 0.001 mg / mL for the n-butanol layer extract and 0.842 ± 0.007 mg / mL and 0.102 ± 0.004 mg / mL for the DPPH free radical scavenging ability of the water layer extract. The results of the FRAP method indicated that it was not based on IC 50The FRAP value of the ethyl acetate layer extract is 262.69 ± 28.13 Trolox equivalents / mg, the FRAP value of the n-butanol layer extract is 491.79 ± 48.10 Trolox equivalents / mg, and the FRAP value of the water layer extract is 100.18 ± 11.16 Trolox equivalents / mg. The present invention can not only provide a theoretical basis for further screening of antioxidant active ingredients in tea fruit camphor and a comparative evaluation method of antioxidant activity, but also provide a certain reference basis for the development of tea fruit camphor products related to cosmetics, food and health care products. The present invention has low cost, simple operation, less time consumption, high efficiency, and good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 The effect of the tea fruit linalool layer extract on SOD (A), CAT (B), GSH (C), and MDA (D); in the figure: Con represents the blank control group, model represents the H2O2 model group, 400, 200, and 100 represent the sample amounts of 400, 200, and 100 μg / mL, respectively. #### represents that compared with the H2O2 group, the blank group p ﹤0.0001, extremely significant; ### represents that compared with the H2O2 group, the blank group p ﹤0.001, significant; **** represents that compared with the H2O2 group, the sample group p ﹤0.0001, *** represents the sample group compared with the H2O2 group p ﹤0.001, ** represents the sample group compared with the H2O2 group p ﹤0.01, * represents the sample group compared with the H2O2 group p ﹤0.05; Figure 2 The effects of the blank control group (A), model group (B) and the high (C), medium (D) and low (E) dose groups of the linalool layer extract of tea fruit on cell apoptosis, as well as the bar graph of cell apoptosis (F). DETAILED DESCRIPTION

[0017] The technical solution of the present invention is further described in detail below in conjunction with the embodiments, but the protection scope of the present invention is not limited thereto.

[0018] Tea fruit camphor ( Cinnamomum chago BS Sun&H. L. Zhao) fruits were collected from Yangbi County, Yunnan.

[0019] In the following examples, unless otherwise specified, ethanol concentrations are expressed in volume percent.

[0020] Example 1 A method for preparing an antioxidant active component of camphor tree, comprising the following steps: 1) Pre-treatment of tea fruit camphor: After peeling off the seed coat and seed shell of fresh Cinnamomum camphora, dry it in the sun (18 h), then crush it with a grinder, pass it through an 80-mesh sieve, and put it into a ziplock bag for later use; 2) Preparation of Cinnamomum camphora crude extract: Accurately weigh 50.0 g of the tea fruit camphor sample obtained after crushing and sieving in step 1) into a 2 L spinner flask, add 75% ethanol aqueous solution according to the material-liquid ratio of 1g (dry sample): 20 mL for water bath heating assisted extraction, the extraction temperature is 50℃, and the extraction time is 2h. After filtering, take the supernatant. Add 75% ethanol aqueous solution to the filter residue and perform water bath heating assisted extraction again in the same way. After the two supernatants are combined and mixed, they are concentrated by vacuum rotary evaporator, and the concentration temperature of the vacuum rotary evaporator is 50℃ to obtain 20.5 g of tea fruit camphor extract. The tea fruit camphor extract is evenly suspended in distilled water (tea fruit camphor total extract and distilled water = 1:1.5, V / V), and extracted with 70 mL of petroleum ether, 70 mL of ethyl acetate, and 70 mL of n-butanol in turn (extracted three times each), respectively obtaining a petroleum ether layer extract, an ethyl acetate layer extract, an n-butanol layer extract, and an aqueous layer extract after n-butanol extraction (hereinafter referred to as the aqueous layer extract).

[0021] Application Experiment 1 The following is a test and evaluation of the antioxidant activity of the ethyl acetate layer extract, the n-butanol layer extract and the water layer extract prepared in Example 1. The DPPH, ABTS and FRAP methods were used to conduct a comprehensive comparative evaluation of the antioxidant activity of the tea fruit camphor samples, respectively. The OD values ​​were measured at wavelengths of 517 nm, 405 nm and 593 nm, respectively, and the DPPH free radical scavenging rate, ABTS free radical scavenging rate and FRAP value were calculated. The antioxidant activity of different extraction layers of tea fruit camphor was compared and evaluated. The antioxidant activity evaluation of the extracts from different extraction layers is shown in Tables 1 to 3.

[0022] ) Determination of DPPH free radical scavenging ability: Preparation of DPPH reagent: 3.943 mg DPPH + 50 mL methanol: Prepare DPPH (M = 394.32, chemical name: 1,1-diphenyl-2-trinitrophenylhydrazine) into a 200 μmol / L solution (prepare and use immediately, valid within 30 minutes).

[0023] Add 10 μL of each sample solution of Cinnamomum camphora (i.e., ethyl acetate layer extract, n-butanol layer extract, and water layer extract) to a 96-well plate, add 175 μL of DPPH reagent, shake and mix, place in a dark place to react for 20 min, and measure the absorbance at 517 nm. The detailed reaction group operation is as follows: Positive drug group: 10 μL positive drug solution (BHA or BHT) + 175 μL DPPH reagent; Positive drug solvent group: 10 μL positive drug solvent (methanol) + 175 μL DPPH reagent; Sample group: 10 μL sample solution + 175 μL DPPH reagent; Sample blank control group: 10 μL sample solvent + 175 μL DPPH reagent; Clearance = (A 空白 -A 样品 ) / A 空白 ×100%.

[0024] ) Determination of ABTS free radical scavenging ability: ABTS preparation: The absorbance was measured by UV spectrophotometer at 0.7-0.8 (at 734 nm).

[0025] K2S2O8 solution: 3.3 mg + 5 mL distilled water; ABTS (2,2-azino-bis(3-ethyl-benzothiazole-6-sulfonic acid) diammonium salt) solution: 134 mg + 3.5 mL distilled water; Mix the above two liquids in a 1:1 volume ratio, let stand for more than 12 hours, and dilute with methanol when using to an absorbance between 0.78-0.82 (405 nm). This can be used as ABTS working solution.

[0026] Add 10 μL of each sample solution of Cinnamomum camphora (i.e., ethyl acetate layer extract, n-butanol layer extract, and water layer extract) to the measurement well, add 200 μL of ABTS solution, react in the dark for 30 min, and measure the absorbance at 405 nm. The detailed reaction group operation is as follows: Positive drug group: 10 μL positive drug solution (BHA or BHT) + 200 μL ABTS working solution; Positive drug solvent group: 10 μL positive drug solvent + 200 μL ABTS working solution; Sample group: 10 μL sample solution + 200 μL ABTS working solution; Sample blank control group: 10 μL sample solvent + 200 μL ABTS working solution.

[0027] Clearance = (A空白 -A 样品 )A 空白 ×100%.

[0028] )FRAP reduction ability assay: Mix the prepared 0.3 mol / L sodium acetate buffer, 20 mol / L ferric chloride solution, and 10 mmol / L TPTZ (chemical name: 2,4,6-tripyridyltriazine) solution in a volume ratio of 10:1:1 to prepare the FRAP working solution (prepare it before use).

[0029] With Trolox (water-soluble vitamin E) concentration as the horizontal axis and its absorbance value as the vertical axis, a Trolox standard curve was drawn (10 mmol / L Trolox ethanol solution was prepared with anhydrous ethanol and diluted to different concentration gradients: 0, 100, 200, 300, 400, 500, 600, 700 and 800 μmo1 / L).

[0030] Sample group: 10 μL sample solution + 200 μL FRAP working solution; Sample blank control group: 10 μL sample solvent + 200 μL FRAP working solution.

[0031] Positive drug group: 10 μL positive drug solution (BHA or BHT prepared into a methanol solution with a concentration of 1 mg / mL) + 200 μL FRAP working solution; Positive drug solvent group: 10 μL positive drug solvent + 200 μL FRAP working solution.

[0032] Repeat each group 3 times.

[0033] Reaction conditions: shake and mix, react at 37℃ in the dark for 30 min, and measure the absorbance at a wavelength of 593 nm. The results are expressed as Trolox equivalents (i.e., the free radical scavenging capacity per gram of sample is equivalent to the number of micromoles of Trolox's free radical scavenging capacity). If the absorbance value of the measured sample exceeds the linear range, the sample needs to be further diluted (the sample can be diluted in different gradients for measurement).

[0034] Table 1. The scavenging ability of different extracts of Cinnamomum camphora samples on DPPH free radicals Table 2. The scavenging ability of different extracts of Cinnamomum camphora samples on ABTS free radicals Table 3. FRAP values ​​of different extracts of Cinnamomum camphora samples Tables 1 to 3 show that the scavenging abilities of the ethyl acetate layer extract on DPPH free radical and ABTS free radical are 0.285 ± 0.006 mg / mL and 0.029 ± 0.008 mg / mL, respectively, the scavenging abilities of the n-butanol layer extract on DPPH free radical and ABTS free radical are 0.208 ± 0.002 mg / mL and 0.008 ± 0.001 mg / mL, respectively, and the scavenging abilities of the water layer extract on DPPH free radical and ABTS free radical are 0.842 ± 0.007 mg / mL and 0.102 ± 0.004 mg / mL, respectively. The FRAP value of the ethyl acetate layer extract is 262.69 ± 28.13 Trolox equivalents / mg, the FRAP value of the n-butanol layer extract is 491.79 ± 48.10 Trolox equivalents / mg, and the FRAP value of the water layer extract is 100.18 ± 11.16 Trolox equivalents / mg. The results in Tables 1 to 3 show that the ethyl acetate layer extract, the n-butanol layer extract and the water layer extract all have good antioxidant activity, among which the n-butanol layer extract has the best antioxidant activity.

[0035] Application Experiment 2 The extract of the n-butanol layer of Cinnamomum camphora was tested for SOD, CAT, GSH, MDA and apoptosis. 1.1 Cell culture 1 × 10 7 HepG2 cells were cultured and 24 hours later, DMEM medium containing the n-butanol layer extract of Cinnamomum camphora was administered (three doses of high, medium and low, 400, 200 and 100 μg / mL). DMEM medium containing 500 mM hydrogen peroxide was administered 24 hours after administration. 4 hours after modeling, the DMEM medium was aspirated, and 1 mL of PBS solution (manufacturer: Servicebio, model: G4202-500 ML) was added to each well to blow the cells. After all the cells were detached from the 6-well plate, the cells in each well were transferred to a 1.5 mL centrifuge tube. Centrifuge at 3000 r / min for 10 min, discard the supernatant, add lysis buffer (TritonX-100 was diluted to 3%-5% with PBS), lyse for 30 min, centrifuge at 12000 r / min for 10 min, and aspirate the supernatant for later use.

[0036] 1 × 10 7HepG2 cells were modeled in DMEM medium without hydrogen peroxide or containing hydrogen peroxide for 4 h, then the DMEM medium was aspirated, 1 mL of PBS solution was added to each well, and the cells were blown. After all the cells were detached from the 6-well plate, the cells in each well were transferred to a 1.5 mL centrifuge tube. Centrifuge at 3000 r / min for 10 min, discard the supernatant, add lysis buffer, lyse for 30 min, centrifuge at 12000 r / min for 10 min, and aspirate the supernatant for use, to obtain the blank group and model group, respectively.

[0037] 1.2 Experimental methods The activities of SOD, CAT, GSH and MDA were determined according to the instructions of the kit.

[0038] 1.3 Experimental Results 1.3.1 SOD test results The total superoxide dismutase (T-SOD) assay kit (WST-1 method) was used to detect the antioxidant activity of the n-butanol layer extract of Cinnamomum camphora. Figure 1 As shown in A, the SOD activity of the blank control group was extremely significant compared with the model group, indicating that hydrogen peroxide induced oxidative stress in HepG2 cells. At concentrations of 400 μg / mL and 200 μg / mL, the SOD activity of the drug-treated group was significantly different from that of the model group, indicating that the extract of the n-butanol layer of Cinnamomum camphora increased the SOD activity of HepG2 cells. The extract of the n-butanol layer of Cinnamomum camphora has a good antioxidant effect.

[0039] 1.3.2 CAT test results The catalase (CAT) assay kit was used to detect the effect of the n-butanol layer extract of Cinnamomum camphora on the CAT activity of HepG2 cells under oxidative stress. Figure 1 As shown in B, the CAT activity of the blank control group was significantly higher than that of the model group, indicating that hydrogen peroxide induced oxidative stress in HepG2 cells. The groups treated with different concentrations of the n-butanol layer extract of Cinnamomum camphora produced significant changes compared with the model group, indicating that the n-butanol layer extract of Cinnamomum camphora could increase the CAT content of HepG2 cells. The n-butanol layer extract of Cinnamomum camphora has a good antioxidant effect.

[0040] 1.3.3 GSH test results The antioxidant activity of the n-butanol layer extract of Cinnamomum camphora was detected by using a reduced glutathione (GSH) assay kit (microplate method). Figure 1As shown in C, the GSH activity of the blank control group was extremely significant compared with the model group, indicating that hydrogen peroxide induced oxidative stress in HepG2 cells. Compared with the model group, the GSH activity of the 100 μg / mL group was significantly increased in the groups treated with different concentrations of the n-butanol layer extract of Cinnamomum camphora, indicating that the n-butanol layer extract of Cinnamomum camphora can increase the GSH content of HepG2 cells. The n-butanol layer extract of Cinnamomum camphora may have a good antioxidant effect.

[0041] 1.3.4 MDA test results The antioxidant activity of the n-butanol layer extract of Cinnamomum camphora was detected by cell malondialdehyde (MDA) assay kit (microplate method). Figure 1 As shown in D, compared with the blank control group, the MDA content in the model group increased significantly, indicating that hydrogen peroxide induced oxidative stress in HepG2 cells. Compared with the model group, the MDA content in the groups treated with different concentrations of the n-butanol layer extract of Cinnamomum camphora was significantly reduced, indicating that the n-butanol layer extract of Cinnamomum camphora could reduce the MDA content of HepG2 cells. The n-butanol layer extract of Cinnamomum camphora may have a good antioxidant effect.

[0042] 1.3.5 Apoptosis detection results Flow cytometry was used to detect the anti-apoptotic ability of the n-butanol layer extract of Cinnamomum camphora. Figure 2 As shown in the figure, compared with the model group, the proportion of live cells in the blank control group decreased from 66.5% to 16.8%, indicating that hydrogen peroxide induced oxidative stress in HepG2 cells, causing large-scale apoptosis of cells. Compared with the model group, the groups treated with different concentrations of the n-butanol layer extract of Cinnamomum camphora inhibited the apoptosis of cells to a certain extent. Among them, the apoptotic cells in the 400 μg / mL group decreased from 72.14% in the model group to 55.24%. This indicates that the n-butanol layer extract of Cinnamomum camphora can inhibit the apoptosis of HepG2 cells caused by hydrogen peroxide to a certain extent, which may indirectly improve the oxidative stress of HepG2 cells induced by H2O2.

[0043] Note: Q4 represents the viable cell population, Q2 and Q3 represent the apoptotic cell population, con represents the control group, model represents the H2O2 model group, 400, 200, and 100 represent the sample volumes of 400, 200, and 100 μg / mL, respectively. ### represents that compared with the H2O2 group, the blank group p ﹤0.001, significant; *** represents that compared with the H2O2 group, the sample group p ﹤0.001, ** represents the sample group compared with the H2O2 group p ﹤0.01, * represents the sample group compared with the H2O2 group p ﹤0.05.

[0044] In summary, the antioxidant active components of the camphor tree of the present invention (i.e., the ethyl acetate layer extract, the n-butanol layer extract and the water layer extract) have good antioxidant activity, and can provide certain theoretical and reference basis for the development of camphor tree products related to cosmetics, food and health care products.

Claims

1. A method for preparing antioxidant active components of Cinnamomum camphora, characterized in that: The steps include: 1) Pre-treatment of tea fruit camphor: After peeling off the seed coat and seed shell of the tea fruit camphor, drying, crushing and setting aside; 2) Preparation of Cinnamomum camphora crude extract: Take the crushed camphor tree, add ethanol aqueous solution and heat at 35-70°C for extraction, separate the solid and liquid after the extraction, concentrate the extract to obtain camphor tree extract, evenly suspend the camphor tree extract in water, extract with petroleum ether, ethyl acetate and n-butanol in sequence, and obtain a petroleum ether layer extract, an ethyl acetate layer extract, an n-butanol layer extract and a water layer extract after n-butanol extraction, respectively; The antioxidant active component of the camphor tree is at least one of an ethyl acetate layer extract, an n-butanol layer extract and an aqueous layer extract after n-butanol extraction.

2. The method for preparing the antioxidant active component of Cinnamomum camphora as claimed in claim 1, characterized in that: In step 1), crush through a 40-100 mesh sieve and put into a ziplock bag for later use.

3. The method for preparing the antioxidant active component of Cinnamomum camphora as claimed in claim 1, characterized in that: In step 2), an ethanol aqueous solution having a volume concentration of 60-85% is added at a solid-liquid ratio of 1 g: 10-30 mL.

4. The method for preparing the antioxidant active component of Cinnamomum camphora as claimed in claim 1, characterized in that: In step 2), the heating extraction time is 2-4 h, and the extraction is performed 1-4 times.

5. The method for preparing the antioxidant active component of Cinnamomum camphora as claimed in claim 1, characterized in that: In step 2), a vacuum rotary evaporator is used for concentration at a concentration temperature of 40-60°C.

6. The method for preparing the antioxidant active component of Cinnamomum camphora as claimed in claim 1, characterized in that: In step 2), the Cinnamomum camphora extract is uniformly suspended in water at a volume ratio of 1:1-3.

7. The antioxidant active component of Cinnamomum camphora obtained by the preparation method according to any one of claims 1 to 6.

8. Use of the antioxidant active components of Cinnamomum camphora as claimed in claim 7 in the preparation of antioxidant drugs.

9. Use of the antioxidant active components of Cinnamomum camphora as claimed in claim 7 in the preparation of drugs for treating or preventing oxidative stress.

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