Artemisia argyi fat-soluble component extraction and determination technology

Abstract

The invention provides a wormwood fat-soluble component extraction and determination process, and belongs to the technical field of fat-soluble extraction.The wormwood fat-soluble component extraction and determination process comprises the steps that moxa is obtained and placed in an extractor for fat-soluble component extraction; obtaining a plurality of parts of fat-soluble components and a plurality of parts of Vc; the total reducing capacity, the hydroxyl free radical scavenging capacity, the DPPH free radical scavenging capacity and the ABTS free radical scavenging capacity of the fat-soluble components are determined, determination of the extracted fat-soluble components of the wormwood is achieved, efficient extraction of the fat-soluble components and determination of the capacity of the fat-soluble components are achieved, and efficient use of the fat-soluble components of the wormwood is achieved.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of fat-soluble extraction, and particularly relates to a fat-soluble component extraction and determination process of wormwood. BACKGROUND

[0002] All life activities in organisms are involved in oxidation-reduction reactions, which maintain physiological activities and homeostasis of the internal environment of organisms. Imbalance of oxidation-reduction reactions can lead to the generation of free radicals and active oxygen atoms, thereby causing irreversible damage to organisms. In recent years, as people's requirements for physical health and food safety are increasingly high, the demand for the development of natural and non-toxic antioxidant products is increasingly high.

[0003] Wormwood (Artemisia argyi Levl.et Vant.) is also known as Xiangai, Aihao, Jiaai, Yicao, Bingtai, Tianai, Jicao, and Hai'ai, and is a one-year or multi-year herb or semi-shrub of the genus Artemisia in the family Asteraceae. Wormwood includes volatile oils, flavonoids, triterpenes and other components, and the whole herb can be used as medicine. The nature and taste of wormwood were first recorded in detail in Tao Hongjing's Shang Bie Lu, which has been used until now. Wormwood has the functions of repelling mosquitoes and insects, dispelling dampness and cold, warming and unblocking meridians, preventing miscarriage and bleeding, and inhibiting bacteria. A large number of studies have shown that a variety of components in wormwood have antioxidant activity, and the antioxidant effect is significant. Different extraction methods have a great influence on the components or effects of wormwood.

[0004] Therefore, a fat-soluble component extraction and determination process of wormwood is needed. SUMMARY

[0005] In view of the above problems, the application provides a fat-soluble component extraction and determination process of wormwood, which comprises the following steps: Obtaining wormwood and placing it in an extractor for fat-soluble component extraction; Obtaining a plurality of portions of fat-soluble components and a plurality of portions of Vc; Determining the total reducing capacity, hydroxyl radical scavenging capacity, DPPH radical scavenging capacity and ABTS radical scavenging capacity of the fat-soluble components.

[0006] Further, the obtaining wormwood and placing it in an extractor for fat-soluble component extraction comprises the following steps: Taking 10 g of wormwood, wrapping it with filter paper in the extractor; The ratio of solid to liquid is 1:10-1:50 g / mL; The extraction temperature is 80-100 DEG C, and the extractor is connected; Extracting for 3-7 hours, and measuring the total flavonoid extraction rate.

[0007] Further, the measurement of the total flavonoid extraction rate comprises the following steps: Plot the standard curve and obtain its equation; Take 1 mL of the extract and place it in a 10 mL stoppered colorimetric tube. Add 0.6 mL of 5% NaNO2 solution, shake well, and let stand for 6 min. Add 10% Mix 0.6 mL of solution, shake well, and let stand for 6 minutes. Add 4 mL of 4% NaOH solution, dilute to volume with 60% ethanol solution, shake well, and let stand for 15 min; The absorbance was measured at a wavelength of 510 nm. The flavonoid content was calculated using a standard curve equation, and a model for calculating the flavonoid extraction rate was established.

[0008] Furthermore, the flavonoid extraction rate calculation model includes: .

[0009] Furthermore, the determination of the total reducing power of the fat-soluble components of Artemisia argyi includes the following steps: The fat-soluble components of Artemisia argyi were diluted with anhydrous ethanol to different concentration gradients, such as 10-100 μL / mL. Take 0.3 mL of the ethanol solution of the fat-soluble components of Artemisia argyi, add 1.5 mL of 0.2 mol / L phosphate buffer solution (pH=6.6), add 1.5 mL of 1% potassium ferricyanide solution and shake well; Place the mixture in a 50°C water bath for 20 minutes, cool it rapidly, and then add 1.5 mL of 10% trichloroacetic acid solution, 0.6 mL of 0.1% ferric chloride solution and 3 mL of distilled water to the mixture in sequence and shake well. After reacting for 10 min, centrifuge at 4000 r / min for 5 min, use anhydrous ethanol as a blank to zero the sample, and measure the absorbance at 700 nm. Use Vc as a positive control (concentration gradient of 0.1-1.2 mg / mL) and compare the absorbance values.

[0010] Furthermore, the determination of the hydroxyl radical scavenging ability of the fat-soluble components of Artemisia argyi includes the following steps: Obtain 1 mL of ethanol solutions of lipophilic components of Artemisia argyi at different concentrations (10-100 μL / mL), add 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of 9 mmol / L salicylic acid solution in sequence, shake well, and let stand for 20 min; measure the absorbance at 510 nm and record it as A1. Take 1 mL of distilled water, add 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of 9 mmol / L salicylic acid solution in turn and shake, stand for 20 min; measure the absorbance at 510 nm and record as A0; Take 1 mL of different concentrations of artemisia argyi fat-soluble component ethanol solution (10-100 μL / mL), add 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of distilled water in turn and shake, stand for 20 min; measure the absorbance at 510 nm and record as A2; Take 1 mL of different concentrations of artemisia argyi fat-soluble component ethanol solution (10-100 μL / mL), add 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of distilled water in turn and shake, stand for 20 min; measure the absorbance at 510 nm and record as A2;

[0011] Further, the hydroxyl radical scavenging rate model comprises: ; Wherein, A1 is the absorbance at 510 nm of 1 mL of different concentrations of artemisia argyi fat-soluble component ethanol solution (10-100 μL / mL), 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of 9 mmol / L salicylic acid solution; A2 is the absorbance at 510 nm of 1 mL of different concentrations of artemisia argyi fat-soluble component ethanol solution (10-100 μL / mL), 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of distilled water; A0 is the absorbance at 510 nm of 1 mL of distilled water, 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of 9 mmol / L salicylic acid solution.

[0012] Further, the determination of the DPPH radical scavenging capacity of the artemisia argyi fat-soluble component comprises the following steps: Take 1 mL of different concentrations of artemisia argyi fat-soluble component ethanol solution (10-100 μL / mL), add 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of distilled water in turn and shake, stand for 20 min; measure the absorbance at 510 nm and record as A2; Take 2 mL of different concentrations of artemisia argyi fat-soluble component ethanol solution, add 2 mL of anhydrous ethanol, shake and measure the absorbance at 517 nm, record as A2; Take 2 mL of DPPH ethanol solution, add 2 mL of anhydrous ethanol, mix and measure the absorbance at 517 nm, record as A0; Take 2 mL of DPPH ethanol solution, add 2 mL of anhydrous ethanol, mix and measure the absorbance at 517 nm, record as A0; Take Vc as a positive control (concentration gradient is 0.001-0.012 mg / mL), and establish a DPPH free radical scavenging rate model.

[0013] Further, the DPPH free radical scavenging rate model comprises: ; Wherein, 2mL of ethanol solution of fat-soluble components of wormwood in different concentration gradients is taken, 2mL of anhydrous ethanol is added, and the absorbance at 517nm is measured as A1; 1mL of ethanol solution of fat-soluble components of wormwood in different concentrations (10-100μL / mL), 1mL of 9mmol / L ferrous sulfate solution, 1mL of 2mmol / L hydrogen peroxide and 1mL of distilled water are taken, and the absorbance at 510nm is recorded as A2; 2mL of DPPH ethanol solution is taken, 2mL of anhydrous ethanol is added, and the absorbance at 517nm is measured as A 0。

[0014] Further, the determination of the DPPH free radical scavenging capacity of the fat-soluble components of wormwood comprises the following steps: Take equal volume of 7mmol / L ABTS solution and 2.45mmol / L potassium persulfate solution, mix at room temperature, and react for 15h in the dark, dilute the mixture with ethanol to make the absorbance at 734nm A 0； The prepared ABTS stock solution can be stored at 4℃ for 3-4 days, 1mL of ethanol solution of wormwood volatile oil in different concentrations (10-100μL / mL) is taken, 3mL of ABTS determination solution is added, mixed, and placed for 20min, the absorbance at 734nm is measured and recorded as A1; Take VC as a positive control (concentration gradient is 0.1-1.2mg / mL); wherein the calculation formula is as follows: .

[0015] The present application extracts fat-soluble components from wormwood, and the total reducing capacity, hydroxyl radical scavenging capacity, DPPH free radical scavenging capacity and ABTS free radical scavenging capacity of the fat-soluble components are determined, so that the determination of the fat-soluble components of wormwood is realized, the fat-soluble components are extracted with high efficiency, the capacity of the fat-soluble components is determined, and the fat-soluble components of wormwood are used efficiently.

[0016] Additional features and advantages of the present application will be set forth in the description that follows, and in part will be apparent from the description, or can be learned by practice of the application. The objectives and other advantages of the present application will be realized and attained by the structure particularly pointed out in the description and appended claims. BRIEF DESCRIPTION OF DRAWINGS

[0017] In order to more clearly illustrate the technical solutions of the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or the prior art description. Obviously, the drawings described below are some embodiments of the present application, and for those skilled in the art, other drawings can also be obtained from these drawings without creative labor.

[0018] Figure 1 The flowchart of the extraction and determination process of the artemisia argyi fat-soluble components in the embodiment of the present application is shown.

[0019] Figure 2 The schematic diagram of the rutin standard curve in the embodiment of the present application is shown.

[0020] Figure 3 The schematic diagram of the influence of temperature on the extraction rate of artemisia argyi fat-soluble components in the embodiment of the present application is shown.

[0021] Figure 4 The schematic diagram of the influence of time on the extraction rate of artemisia argyi fat-soluble components in the embodiment of the present application is shown.

[0022] Figure 5 The schematic diagram of the influence of the ratio of material to liquid on the extraction rate of artemisia argyi fat-soluble components in the embodiment of the present application is shown.

[0023] Figure 6 The schematic diagram of the total reducing capacity of Vc in the embodiment of the present application is shown.

[0024] Figure 7 The schematic diagram of the total reducing capacity of artemisia argyi fat-soluble components in the embodiment of the present application is shown.

[0025] Figure 8 The schematic diagram of the scavenging rate of hydroxyl radicals by Vc in the embodiment of the present application is shown.

[0026] Figure 9 The schematic diagram of the scavenging rate of hydroxyl radicals by artemisia argyi fat-soluble components in the embodiment of the present application is shown.

[0027] Figure 10 The schematic diagram of the scavenging rate of DPPH radicals by Vc in the embodiment of the present application is shown.

[0028] Figure 11 The schematic diagram of the scavenging rate of DPPH radicals by artemisia argyi fat-soluble components in the embodiment of the present application is shown.

[0029] Figure 12 A schematic diagram of the ABTS free radical scavenging rate of Vc in the embodiment of the present application is shown.

[0030] Figure 13 A schematic diagram of the ABTS free radical scavenging rate of wormwood fat-soluble in the embodiment of the present application is shown. DETAILED DESCRIPTION

[0031] In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are some but not all of the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the protection scope of the present application.

[0032] Reference Figure 1 A wormwood fat-soluble component extraction and determination process, comprising the following steps: Obtaining mink oil and placing it in an extractor for fat-soluble component extraction; Obtaining several portions of wormwood fat-soluble components and several portions of Vc (Vitamin C); Determining the total reducing capacity, hydroxyl radical scavenging capacity, DPPH free radical scavenging capacity and ABTS free radical scavenging capacity of the fat-soluble components.

[0033] Step 1, establishing a standard curve Accurately weighing 10.00 mg of rutin standard, adding an appropriate amount of 60% ethanol solution, heating and dissolving in a 50°C water bath, naturally cooling, and diluting to 50 mL with 60% ethanol solution to obtain a rutin standard solution with a concentration of 0.20 mg / mL. Accurately pipetting 0.0 mL, 1.0 mL, 2.0 mL, 3.0 mL, 4.0 mL and 5.0 mL of the rutin standard solution into 10 mL stoppered cuvettes. Adding 0.6 mL of 5% NaNO2 solution, shaking well, and standing for 6 min, then adding 0.6 mL of 10% Al(NO3)3 solution, shaking well, and standing for 6 min. Adding 4 mL of 4% NaOH solution, shaking well, and standing for 15 min. Measuring the absorbance at a wavelength of 510 nm. Plotting the standard curve (as shown in FIG. 1) with the flavonoid content as the abscissa and the absorbance as the ordinate. Figure 2 Figure 2 Calculating the regression equation: Y=8.7509X, and the correlation coefficient R2=0.9999. In the formula, Y is the absorbance value, and X is the rutin concentration (mg / mL).

[0034] ​​Step 2, Determination of total flavonoid content in Artemisia argyi Accurately transfer 1 mL of the extract into a 10 mL stoppered colorimetric tube. Add 0.6 mL of 5% NaNO₂ solution, shake well, let stand for 6 min, then add 10% NaNO₂ solution. Add 0.6 mL of solution, shake well, let stand for 6 min, add 4 mL of 4% NaOH solution, dilute to volume with 60% ethanol solution, shake well, and let stand for 15 min. Measure the absorbance at 510 nm. Calculate the flavonoid content using the standard curve equation, and then calculate the flavonoid extraction rate. Repeat each experiment three times.

[0035] The extraction rate is calculated using the following formula: .

[0036] The following experiments investigated the effects of different conditions on the extraction rate of fat-soluble components from Artemisia argyi. (1) Effect of extraction temperature on the extraction rate of fat-soluble components of Artemisia argyi Accurately weigh 10g of Artemisia flakes, wrap them in filter paper and place them in a Soxhlet extractor. The material-to-liquid ratio is 1:30g / mL (i.e., 300mL of anhydrous ethanol). The extraction temperatures are 80, 85, 90, 95, and 100℃. Connect the Soxhlet extractor and extract for 3 hours. Measure the total flavonoid extraction rate.

[0037] Depend on Figure 3 As shown in Table 1, the extraction rate increases with increasing temperature at lower extraction temperatures, reaching its highest point at 95℃ (13.706 mg / g), exhibiting highly significant differences compared to 80, 85, and 90℃, and a significant difference compared to 100℃. Further increases in temperature lead to a decrease in extraction rate, possibly due to partial decomposition of flavonoids at excessively high temperatures. Therefore, controlling the extraction temperature between 90 and 100℃ is recommended. An extraction temperature of 95℃ was selected.

[0038] Table 1. Effect of extraction temperature on the extraction rate of fat-soluble components from Artemisia argyi.

[0039] (2) Effect of extraction time on the extraction rate of fat-soluble components of Artemisia argyi Accurately weigh 10g of Artemisia flakes, wrap them in filter paper and place them in a Soxhlet extractor. The material-to-liquid ratio is 1:30g / mL. The extraction temperature is 95℃, and the Soxhlet extraction times are 3, 4, 5, 6 and 7 hours, respectively. Measure the total flavonoid content.

[0040] Depend on Figure 4As shown in Table 2, with the increase of extraction time, the extraction rate first increases and then decreases. The extraction rate is the largest when the extraction time is 5h, which is 22.161mg / g, and there is a significant difference compared with other extraction times. When the extraction time is greater than 5h, the flavonoids are partially decomposed due to the high temperature extraction for a long time, so the extraction rate decreases. Therefore, the extraction time is preferably controlled between 4-6h. The extraction time is selected as 5h.

[0041] Table 2 Effect of extraction time on the extraction rate of fat-soluble components of wormwood

[0042] (3) Effect of solid-liquid ratio on the extraction rate of fat-soluble components of wormwood Accurately weigh 10g of wormwood, wrap the filter paper in the Soxhlet extractor, the solid-liquid ratio is 1:10, 1:20, 1:30, 1:40, 1:50g / mL, the extraction temperature is 95℃, the Soxhlet extraction is 3h, and the total flavonoid content is measured.

[0043] Specifically, by Figure 5 As shown in Table 3, with the increase of solid-liquid ratio from 1:10 to 1:30, the extraction rate increases greatly, and there is a significant difference. When the solid-liquid ratio is from 1:30 to 1:50, the extraction rate increases slightly, the highest extraction rate is 14.693mg / g, the solvent consumption is small, the extraction is not complete, but the solvent consumption is large, and the effect is not obvious. Therefore, from the experimental and economic points of view, the solid-liquid ratio is preferably controlled between 1:25-1:35. The solid-liquid ratio is selected as 1:30.

[0044] Table 3 Effect of solid-liquid ratio on the extraction rate of fat-soluble components of wormwood

[0045] The following experiments are conducted to determine the fat-soluble component capacity of wormwood Determination of total reducing capacity of fat-soluble components of wormwood The artemisia argyi liposoluble components were diluted with anhydrous ethanol to different concentration gradients of 10, 20, 30, 40, 60, 80 and 100 μL / mL. 0.3 mL of artemisia argyi liposoluble component ethanol solution was added to 1.5 mL of 0.2 mol / L phosphate buffer solution (PH = 6.6), 1.5 mL of 1% potassium ferricyanide solution was added, and the mixture was shaken and water bathed in a 50°C water bath for 20 min, then quickly cooled, and then 1.5 mL of 10% trichloroacetic acid solution, 0.6 mL of 0.1% ferric trichloride solution and 3 mL of distilled water were added in turn, shaken, reacted for 10 min and then centrifuged at 4000 r / min for 5 min. The anhydrous ethanol was used as a blank to zero, the absorbance at 700 nm was measured, and Vc was used as a positive control (concentration gradient of 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2 mg / mL). The higher the absorbance value, the stronger the total reducing capacity.

[0046] Specifically, Figure 6 Vc and Figure 7 The total reducing capacity of artemisia argyi liposoluble components is shown in the figure. As shown in the figure, Vc and artemisia argyi liposoluble components both have increasing reducing capacity with increasing concentration. There are some differences in the total reducing capacity of the two, and the reducing capacity of Vc is significantly higher than that of artemisia argyi liposoluble components, but when the concentration of artemisia argyi liposoluble components reaches 100 μL / mL, it also has a certain reducing capacity.

[0047] Determination of the hydroxyl radical scavenging capacity of artemisia argyi liposoluble components 1 mL of artemisia argyi liposoluble component ethanol solution (10, 20, 30, 40, 60, 80 and 100 μL / mL) was taken, 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of 9 mmol / L salicylic acid solution were added in turn, shaken, allowed to stand for 20 min, and the absorbance at 510 nm was measured and recorded as A1; 1 mL of distilled water was used instead of artemisia argyi liposoluble component ethanol solution, and the rest of the conditions were unchanged, and the absorbance at 510 nm was measured and recorded as A0; 1 mL of distilled water was used instead of the above salicylic acid solution, and the rest of the conditions were unchanged, and the absorbance at 510 nm was measured and recorded as A 2， VC was used as a positive control (concentration gradient of 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2 mg / mL), each group of tests was repeated 3 times, and the hydroxyl radical scavenging rate was calculated according to the following formula.

[0048] .

[0049] Specifically, Figure 8 Vc and Figure 9The scavenging ability of the fat-soluble components of Artemisia argyi against hydroxyl radicals was investigated. As shown in the figure, within the oil concentration range of 10–100 μL / mL, the scavenging rate of the fat-soluble components gradually increased with increasing concentration. When the concentration of the fat-soluble components reached 100 μL / mL, the scavenging rate of hydroxyl radicals was 83.76%, which was 1.47 times that of 0.2 mg / mL. These results indicate that the fat-soluble components of Artemisia argyi have a significant ability to scavenge hydroxyl radicals.

[0050] Determination of the scavenging ability of fat-soluble components of Artemisia argyi against DPPH free radicals Prepare a 0.1 mmol / L DPPH ethanol solution and store it in a brown bottle for later use. Take 2 mL of ethanol solutions containing different concentration gradients of the fat-soluble components of Artemisia argyi (10, 20, 30, 40, 60, 80, and 100 μL / mL), add 2 mL of DPPH ethanol solution, shake well, and react in the dark for 20 min. Measure the absorbance at 517 nm and record it as A1. Take another 2 mL of ethanol solutions containing different concentration gradients of the fat-soluble components of Artemisia argyi, add 2 mL of anhydrous ethanol, shake well, and measure the absorbance at 517 nm, record it as A2. Take 2 mL of DPPH ethanol solution, add 2 mL of anhydrous ethanol, mix well, and measure the absorbance at 517 nm, record it as A3. 0。 Vitamin C was used as a positive control (concentration gradient of 0.001, 0.002, 0.004, 0.006, 0.008, 0.01, and 0.012 mg / mL). Each experiment was repeated three times, and the DPPH free radical scavenging rate was calculated using the following formula.

[0051] .

[0052] Figure 10 For Vc and Figure 11 The figure above shows the scavenging ability of the fat-soluble components of Artemisia argyi against DPPH free radicals. Both vitamin C and the fat-soluble components exhibit good scavenging ability against DPPH free radicals. At a concentration of 40 μL / mL, the scavenging rate of DPPH free radicals is 92.19%, which is 1.03 times that of 0.010 mg / mL Vc. When the concentration is greater than 40 μL / mL, the scavenging ability is relatively stable, with scavenging rates approaching 100%. These results indicate that the fat-soluble components of Artemisia argyi have a significant DPPH free radical scavenging ability.

[0053] Determination of the ability of fat-soluble components of Artemisia argyi to scavenge ABTS free radicals Take equal volume of 7mmol / L ABTS solution and 2.45mmol / L potassium persulfate solution mixture, at room temperature, avoid light reaction 15h, diluted with ethanol mixture to make its absorbance at wavelength of 734nm A0. Configuration of ABTS stock solution can be stored at 4℃ for 3~4 days. Take 1mL different concentrations of wormwood volatile oil ethanol solution (10, 20, 30, 40, 60, 80 and 100μL / mL), add 3mL ABTS assay solution, mix, stand for 20min, determine its absorbance at 734nm, and record as A1, VC as positive control (concentration gradient is 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2mg / mL), each group of test repeated 3 times, according to the following formula to calculate ABTS free radical scavenging rate.

[0054] .

[0055] ABTS free radical scavenging capacity of wormwood liposoluble components Figure 12 Vc and Figure 13 ABTS free radical scavenging capacity of wormwood liposoluble components, from the above figure, Vc and wormwood liposoluble components have strong ABTS free radical scavenging capacity, and show a dose-effect relationship, Vc greater than 0.4mg / mL, the free radical scavenging capacity is stable, the clearance rate is close to 100%. When the concentration of wormwood liposoluble components is 100μL / mL, the clearance rate of ABTS free radical is as high as 97.97%, the free radical scavenging capacity is 1.35 times of 0.2mg / mL Vc, which is equivalent to the scavenging capacity of 0.4mg / mL Vc. The results show that wormwood liposoluble components have significant ABTS free radical scavenging capacity.

[0056] From the above experiment, we can know (1) Soxhlet extraction method was used to extract wild wormwood liposoluble components, through single factor experiment, the best extraction process was determined as follows: extraction temperature was 95℃, extraction time was 5h, and solid-liquid ratio was 1∶30.

[0057] (2) The total reducing capacity of wild wormwood liposoluble components and the clearance rate of DPPH free radical, hydroxyl radical and ABTS free radical were positively correlated with the concentration, and had significant scavenging capacity for the three kinds of free radicals, which confirmed that wild wormwood liposoluble components had good antioxidant activity. Wild wormwood can be used to develop natural antioxidant products.

[0058] Although the present application has been described in detail with reference to the foregoing embodiments, it should be understood that modifications can be made to the foregoing embodiments, or additional implementations of the present application can be implemented, without departing from the spirit or scope of the application. Accordingly, the present application is not limited except as by the appended claims.

Claims

1. A process for extracting and determining the fat-soluble components of Artemisia argyi, characterized in that, Includes the following steps: The mugwort floss was obtained and placed in an extractor for extraction of fat-soluble components; Obtain a certain amount of fat-soluble components and a certain amount of vitamin C; The total reducing power, hydroxyl radical scavenging power, DPPH radical scavenging power, and ABTS radical scavenging power of the lipid-soluble components were determined.

2. The process for extracting and determining the fat-soluble components of Artemisia argyi according to claim 1, characterized in that, The process of obtaining mugwort floss and placing it in an extractor for fat-soluble component extraction includes the following steps: Take 10g of mugwort floss, wrap it in filter paper, and place it in the extractor; The material-to-liquid ratio is 1:10-1:50 g / mL; The extraction temperatures are 80-100℃; connect the extractor. Extract for 3-7 hours and measure the total flavonoid extraction rate.

3. The process for extracting and determining the fat-soluble components of Artemisia argyi according to claim 1, characterized in that, The determination of its total flavonoid extraction rate includes the following steps: Plot the standard curve and obtain its equation; Take 1 mL of the extract into a 10 mL stoppered colorimetric tube; add 0.6 mL of 5% NaNO2 solution, shake well, and let stand for 6 min; Add 10% Mix 0.6 mL of solution, shake well, and let stand for 6 minutes; Add 4 mL of 4% NaOH solution, dilute to volume with 60% ethanol solution, shake well, and let stand for 15 min; The absorbance was measured at a wavelength of 510 nm. The flavonoid content was calculated using a standard curve equation, and a model for calculating the flavonoid extraction rate was established.

4. The process for extracting and determining the fat-soluble components of Artemisia argyi according to claim 3, characterized in that, The flavonoid extraction rate calculation model includes: 。 5. The process for extracting and determining the fat-soluble components of Artemisia argyi according to claim 1, characterized in that, The determination of the total reducing power of fat-soluble components in Artemisia argyi includes the following steps: The fat-soluble components of Artemisia argyi were diluted with anhydrous ethanol to different concentration gradients, such as 10-100 μL / mL. Take 0.3 mL of the ethanol solution of the fat-soluble components of Artemisia argyi, add 1.5 mL of 0.2 mol / L phosphate buffer solution (pH=6.6), add 1.5 mL of 1% potassium ferricyanide solution and shake well; Place the mixture in a 50°C water bath for 20 minutes, cool it rapidly, and then add 1.5 mL of 10% trichloroacetic acid solution, 0.6 mL of 0.1% ferric chloride solution and 3 mL of distilled water to the mixture in sequence and shake well. After reacting for 10 min, centrifuge at 4000 r / min for 5 min, use anhydrous ethanol as a blank to zero the sample, and measure the absorbance at 700 nm. Use Vc as a positive control (concentration gradient of 0.1-1.2 mg / mL) and compare the absorbance values.

6. The process for extracting and determining the fat-soluble components of Artemisia argyi according to claim 1, characterized in that, The determination of the hydroxyl radical scavenging ability of lipid-soluble components includes the following steps: Obtain 1 mL of ethanol solutions of lipophilic components of Artemisia argyi at different concentrations (10-100 μL / mL), add 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of 9 mmol / L salicylic acid solution in sequence, shake well and let stand for 20 min to react. Its absorbance at 510 nm was measured and recorded as A1; Take 1 mL of distilled water, add 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of 9 mmol / L salicylic acid solution in sequence and shake well, let stand for 20 min to react; Its absorbance at 510 nm was measured and recorded as A0; Obtain 1 mL of ethanol solutions of lipophilic components of Artemisia argyi at different concentrations (10-100 μL / mL), add 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of distilled water in sequence, shake well and let stand for 20 min to react; Its absorbance at 510 nm was measured and recorded as A2; A hydroxyl radical scavenging model was established using vitamin C as a positive control (concentration gradient of 0.1-1.2 mg / mL).

7. The process for extracting and determining the fat-soluble components of Artemisia argyi according to claim 1, characterized in that, Hydroxyl radical scavenging models include: ； Where, A1 is the absorbance at 510 nm of 1 mL of ethanol solution (10-100 μL / mL) of 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide solution and 1 mL of 9 mmol / L salicylic acid solution. A2 represents the absorbance at 510 nm of 1 mL of ethanol solutions (10-100 μL / mL) of the fat-soluble components of Artemisia argyi at different concentrations, 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of distilled water. A0 is the absorbance at 510 nm of 1 mL distilled water, 1 mL 9 mmol / L ferrous sulfate solution, 1 mL 2 mmol / L hydrogen peroxide, and 1 mL 9 mmol / L salicylic acid solution.

8. The process for extracting and determining the fat-soluble components of Artemisia argyi according to claim 1, characterized in that, The determination of the DPPH free radical scavenging ability of the fat-soluble components of Artemisia argyi includes the following steps: Obtain a 0.1 mmol / L DPPH ethanol solution and store it in a brown bottle for later use; Take 2 mL of ethanol solutions of fat-soluble components of Artemisia argyi at different concentration gradients (10-100 μL / mL), add 2 mL of DPPH ethanol solution, shake well, react in the dark for 20 min, and measure the absorbance at 517 nm, which is recorded as A1. Take another 2 mL of ethanol solution of fat-soluble components of Artemisia argyi at different concentration gradients, add 2 mL of anhydrous ethanol, shake well, and measure its absorbance at 517 nm, which is recorded as A2. Take 2 mL of DPPH ethanol solution, add 2 mL of anhydrous ethanol, mix well, and measure its absorbance at 517 nm, which is recorded as A0. A DPPH free radical scavenging model was established using vitamin C as a positive control (concentration gradient of 0.001-0.012 mg / mL).

9. The process for extracting and determining the fat-soluble components of Artemisia argyi according to claim 1, characterized in that, The DPPH radical scavenging rate model includes: ； Among them, 2 mL of ethanol solutions of fat-soluble components of Artemisia argyi at different concentration gradients were taken, 2 mL of anhydrous ethanol was added, the mixture was shaken well, and the absorbance at 517 nm was measured as A1. The absorbance at 510 nm of 1 mL of ethanol solution (10-100 μL / mL) of fat-soluble components of Artemisia argyi at different concentrations, 1 mL of 9 mmol / L ferrous sulfate solution, 1 mL of 2 mmol / L hydrogen peroxide and 1 mL of distilled water is recorded as A2. Add 2 mL of anhydrous ethanol to 2 mL of DPPH ethanol solution, mix well, and measure the absorbance at 517 nm, denoted as A. 0。 10. The process for extracting and determining the fat-soluble components of Artemisia argyi according to claim 1, characterized in that, The determination of the DPPH free radical scavenging ability of the fat-soluble components of Artemisia argyi includes the following steps: Mix equal volumes of 7 mmol / L ABTS solution and 2.45 mmol / L potassium persulfate solution, and react at room temperature in the dark for 15 h. Dilute the mixture with ethanol to achieve an absorbance of A at a wavelength of 734 nm. 0； The prepared ABTS stock solution can be stored at 4℃ for 3-4 days. Take 1 mL of ethanol solution of Artemisia argyi volatile oil of different concentrations (10-100 μL / mL), add 3 mL of ABTS test solution, mix well, let stand for 20 min, measure its absorbance at 734 nm, and record it as A1. Vitamin C was used as a positive control (concentration gradient of 0.1-1.2 mg / mL); the calculation formula is as follows:

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