Purple jade extract as well as preparation method and application thereof

Abstract

The invention relates to a purple jade extract as well as a preparation method and application thereof, and belongs to the field of plant extracts. According to the purple jade extract as well as the preparation method and the application thereof provided by the invention, the extraction of flavonoid substances rich in a dendrobium devonianum variety'purple jade 'is completed for the first time, the extract with the total flavonoid content of more than or equal to 17% is obtained, and the condition that the dendrobium devonianum variety'purple jade' contains two substances, namely rutin and vicenin-2, is determined through qualitative and quantitative analysis. Through verification, the purple jade extract disclosed by the invention has a good antioxidant function, and the total antioxidant capacity of the purple jade extract is close to that of a water-soluble vitamin E analogue Trolox (6-hydroxy-2, 5, 7, 8-tetramethylchromane-2-carboxylic acid) with the equivalent weight of 4mM. According to the purple jade extract as well as the preparation method and the application thereof, a scientific basis is provided for application of purple jade in related directions, and the purple jade extract can be used as an intermediate for development of purple jade series products (such as in the fields of medicine, food, daily necessity development and the like).

Description

Technical Field

[0001] The invention belongs to the field of plant extracts, and particularly relates to a purple jade extract and a preparation method and application thereof. Background Art

[0002] Free radicals, also known as free radicals, are products of chemical reactions related to natural metabolism. Excessive free radicals can disrupt the body's defenses, causing oxidative stress and significant harm to the human body. This is because free radicals are atoms, molecules, or ions with unpaired electrons. They easily react with other molecules, stealing electrons and altering their structure. This can lead to a range of diseases and even cancer. Antioxidants and antioxidant enzymes can scavenge excess free radicals and prevent oxidative stress. The search for substances with high antioxidant activity has long been a persistent goal of scientific research.

[0003] Dendrobium devonianum Paxton (also known as purple-skinned dendrobium, scientific name: Dendrobium devonianum Paxton) is a perennial herbaceous plant of the genus Dendrobium in the Orchidaceae family. It is a geographical indication product of Baoshan City, Yunnan Province, and is commonly used in folk medicine. As a similar product to Dendrobium officinale, it has the benefits of nourishing yin and promoting fluid production; moistening the lungs and kidneys; and strengthening the stomach and intestines. Modern pharmacological research has shown that Dendrobium devonianum Paxton has hypoglycemic, immunomodulatory, and anti-inflammatory activities. Studies have found that Dendrobium devonianum mainly contains chemical components such as polysaccharides, flavonoids, phenanthrenes, and lignin. Among them, the significant antioxidant capacity of flavonoids cannot be ignored. They can inhibit the formation of free radicals by binding to oxidases, complexing with metal ions, and upregulating the expression of antioxidant enzymes. They can also directly scavenge peroxides and free radicals, maintaining the body's balance.

[0004] Ziyu, a new horticultural variety of Dendrobium dentata approved by the Yunnan Provincial Forestry and Grassland Bureau in 2023, originates from Longling County, Baoshan City, Yunnan Province. This plant boasts a distinctive deep purple hue throughout, distinguishing it from other Dendrobium dentata varieties (which have purple outer skin and green mid-stems). However, Ziyu has yet to be used to extract products with high antioxidant activity. Summary of the Invention

[0005] The present invention aims to provide a Ziyu extract, its preparation method, and its application. This invention achieves the first extraction of flavonoid-rich substances from the Dendrobium officinale variety "Ziyu," resulting in a Ziyu extract with a total flavonoid content of ≥17%. Qualitative and quantitative analysis confirms that the extract contains rutin and vetamine-2, two substances with excellent antioxidant properties, with a total antioxidant capacity approaching that of the water-soluble vitamin E analog Trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid), equivalent to 4 mM.

[0006] The present invention provides a method for preparing a purple jade (Yunlin Garden Plant New Registration No. 20230032) extract, comprising the following steps:

[0007] The purple jade is extracted with ethanol aqueous solution as solvent to obtain the purple jade extract.

[0008] Preferably, the ethanol aqueous solution contains 80% to 95% ethanol by volume.

[0009] Preferably, after the extraction, the process further comprises filtering the filtrate, concentrating and drying.

[0010] Preferably, before the extraction, the purple jade is crushed, and the particle size of the crushed jade is 20 to 60 meshes.

[0011] The present invention also provides a purple jade extract prepared by the preparation method described in the above technical solution.

[0012] Preferably, the purple jade extract contains flavonoids; the flavonoids include rutin and vinblastine-2.

[0013] The present invention also provides a purple jade extract prepared by the preparation method described in the above technical solution, or the present invention also provides the use of the purple jade extract described in the above technical solution in the preparation of antioxidant products.

[0014] The present invention also provides a purple jade extract prepared by the preparation method described in the above technical solution, or the present invention also provides the use of the purple jade extract described in the above technical solution in the preparation of antioxidant foods.

[0015] The present invention also provides a purple jade extract prepared by the preparation method described in the above technical solution, or the present invention also provides the use of the purple jade extract described in the above technical solution in the preparation of antioxidant drugs.

[0016] The present invention also provides a purple jade extract prepared by the preparation method described in the above technical solution, or the present invention also provides the use of the purple jade extract described in the above technical solution in the preparation of daily chemical products.

[0017] The present invention provides an extract and application of Dendrobium officinale (Ziyu). This study, for the first time, has extracted flavonoid-rich substances from the Dendrobium officinale variety "Ziyu," yielding an extract with a total flavonoid content of ≥17%. Qualitative and quantitative analysis also confirmed the presence of rutin and vecinin-2 in this Dendrobium officinale variety.

[0018] The Dendrobium candidum-Purple Jade extract of the present invention has been verified to have excellent antioxidant function, with a total antioxidant capacity close to that of 4mM equivalent of the water-soluble vitamin E analog Trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid). It not only has excellent hydroxyl radical scavenging ability, superoxide anion inhibition ability, and superoxide dismutase (SOD) inhibition ability, but also has a certain DPPH free radical scavenging ability. The purple jade extract of the present invention, its preparation method, and application provide a scientific basis for the application of purple jade in related areas, and can be used as an intermediate in the development of purple jade products (such as in the fields of medicine, food, and daily necessities development). BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0020] Figure 1 The HPLC spectrum of the purple jade extract provided by the present invention;

[0021] Figure 2 The mass spectrum of vecinin-2 and rutin contained in the purple jade provided by the present invention (A is vecinin-2, B is rutin);

[0022] Figure 3 This is a comparison chart of the total antioxidant capacity of the purple jade extract, rutin and vecinin-2 provided by the present invention (the extract concentration is 0.25 mg / mL and the flavonoid concentration is 25 μM);

[0023] Figure 4 A comparison chart of the hydroxyl radical scavenging abilities of the purple jade extract, rutin, and vecinin-2 provided by the present invention (the extract concentration is 0.25 mg / mL and the flavonoid concentration is 25 μM);

[0024] Figure 5 A comparison chart of the superoxide anion inhibition abilities of the purple jade extract, rutin, and vecinin-2 provided by the present invention (the extract concentration is 0.25 mg / mL and the flavonoid concentration is 25 μM);

[0025] Figure 6 A comparison chart of the DPPH free radical scavenging abilities of the purple jade extract, rutin, and vecinin-2 provided by the present invention (the extract concentration is 0.25 mg / mL and the flavonoid concentration is 25 μM);

[0026] Figure 7This is a comparison chart of the SOD scavenging abilities of the purple jade extract, rutin, and vecinin-2 provided by the present invention (the extract concentrations are 0.125 mg / mL, 0.25 mg / mL, 0.5 mg / mL, 1 mg / mL, and 2 mg / mL, and the flavonoid concentrations are 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, and 50 μM). DETAILED DESCRIPTION

[0027] The present invention provides a method for preparing a purple jade (Yunlin Garden Plant New Registration No. 20230032) extract, comprising the following steps:

[0028] The purple jade is extracted with ethanol aqueous solution as solvent to obtain the purple jade extract.

[0029] In a specific embodiment, the extraction further comprises crushing the purple jade, wherein the crushed particle size is 20 to 60 mesh. In a specific embodiment, the purple jade can be the stem of the purple jade plant.

[0030] In a specific embodiment, the ethanol aqueous solution contains 80% to 95% ethanol by volume. In a specific embodiment, the extraction time can be 24 hours.

[0031] In a specific embodiment, after the extraction, the process further includes filtering the filtrate, concentrating and drying. In a specific embodiment, the filtration operation can be performed using filter paper. In a specific embodiment, the filtration using filter paper can be repeated 3 times. In a specific embodiment, the filtration operation is repeated 3 times, and the filtrates are combined 3 times. In a specific embodiment, the concentration and drying can be performed using a rotary evaporator to concentrate and dry under reduced pressure. In a specific embodiment, the purple jade extract can be concentrated and dried until the sample becomes solid.

[0032] The present invention also provides a Ziyu extract prepared by the preparation method described in the above technical solution. The present invention completes the extraction of flavonoid-rich substances from the Dendrobium densiflorum variety "Ziyu" for the first time and obtains an extract with a total flavonoid content of ≥17%.

[0033] In a specific embodiment, the purple jade extract contains flavonoids; the flavonoids include rutin and venetian-2.

[0034] The present invention also provides a purple jade extract prepared by the preparation method described in the above technical solution, or the present invention also provides the application of the purple jade extract described in the above technical solution in the preparation of antioxidant products. In a specific embodiment, it has been verified that the Dendrobium candidum-purple jade extract of the present invention has good antioxidant function, and the total antioxidant capacity is close to 4mM equivalent of the water-soluble vitamin E analog Trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid). Among them, not only the hydroxyl radical scavenging ability, superoxide anion inhibition ability, and superoxide dismutase (SOD) inhibition ability are very excellent, but also it has a certain DPPH free radical scavenging ability. The purple jade extract described in the present invention, its preparation method and application provide a scientific basis for the application of purple jade in related directions.

[0035] The present invention also provides a purple jade extract prepared by the preparation method described in the above technical solution, or the present invention also provides the use of the purple jade extract described in the above technical solution in the preparation of antioxidant foods.

[0036] The present invention also provides a purple jade extract prepared by the preparation method described in the above technical solution, or the present invention also provides the use of the purple jade extract described in the above technical solution in the preparation of antioxidant drugs.

[0037] The present invention also provides a purple jade extract prepared by the preparation method described in the above technical solution, or the present invention also provides the use of the purple jade extract described in the above technical solution in the preparation of daily chemical products.

[0038] In order to further illustrate the present invention, a purple jade extract provided by the present invention, its preparation method and application are described in detail below with reference to the accompanying drawings and examples, but they should not be understood as limiting the scope of protection of the present invention.

[0039] The sources of some raw materials in this application are as follows; other experimental reagents not mentioned can be purchased through conventional commercial channels.

[0040] Source of purple jade raw materials: Longling Dendrobium Research Institute, Longling County, Baoshan City, Yunnan Province.

[0041] Example 1

[0042] Preparation of Ziyu Extract and Determination of Flavonoids Content.

[0043] 1. Reagents.

[0044] Rutin, alcohol extract of purple jade, anhydrous ethanol, methanol, sodium nitrite, aluminum nitrate, sodium hydroxide, purified water.

[0045] 2. Experimental method.

[0046] Take 250g of the raw material of purple jade, completely crush it, and soak it in 95% ethanol for 24 hours. Filter it through filter paper. Repeat this process three times and combine the filtrates. Spin dry the liquid and collect the remaining solid in the flask to obtain the purple jade alcohol extract.

[0047] Accurately weigh an appropriate amount of rutin standard into a 1.5mL EP tube, add the corresponding volume of methanol to dissolve, and prepare a standard solution with a concentration of 1.0mg / mL. Dilute the standard solution into a series of gradient concentration solutions (0.1mg / mL, 0.2mg / mL, 0.4mg / mL, 0.8mg / mL, 1.0mg / mL). The sample alcohol extract is also prepared into a 1.0mg / mL standard solution. Take 0.5mL of each and place it in a 10mL volumetric flask, add 0.4mL of 5% sodium nitrite; after standing for 6 minutes, add 0.4mL of 10% aluminum nitrate; after standing for 6 minutes, add 4mL of 4% sodium hydroxide, add anhydrous ethanol to the scale line, shake well; after standing for 15 minutes, measure the absorbance at a wavelength of 510nm. Draw a standard curve based on the absorbance of the gradient solution, and use the standard curve (y=0.9836x-0.0328, R 2 =0.9985) to calculate the content of flavonoids in the sample.

[0048] 3. Experimental results.

[0049] The HPLC spectrum of the purple jade extract was obtained as follows Figure 1 According to the determination results, the yield of the alcohol extract of purple jade is 3.39%, of which the flavonoid content is 17.51%.

[0050] Example 2

[0051] Comparison of flavonoid content of Ziyu extract with other Dendrobium denticulate extracts.

[0052] 1. Reagents.

[0053] Rutin, anhydrous ethanol, methanol, sodium nitrite, aluminum nitrate, sodium hydroxide, purified water.

[0054] 2. Experimental method.

[0055] 250g each of eight different varieties of Dendrobium officinale (Dendrobium officinale No. 1, Dendrobium officinale No. 2, Longzi No. 1, Longzi No. 2, Longzi No. 4, Longzi No. 5, Ziyu, and Wujincao) were collected and completely ground. The mixture was then soaked in 95% ethanol for 24 hours and filtered through filter paper. This was repeated three times, and the filtrates were combined. The samples were concentrated and dried under reduced pressure on a rotary evaporator until they became solid, thus obtaining the alcohol-extracted fraction of Dendrobium officinale.

[0056] Accurately weigh an appropriate amount of rutin standard into a 1.5mL EP tube, add the corresponding volume of methanol to dissolve, and prepare a standard solution with a concentration of 1.0mg / mL. Dilute the standard solution into a series of gradient concentration solutions (0.1mg / mL, 0.2mg / mL, 0.4mg / mL, 0.8mg / mL, 1.0mg / mL). The sample alcohol extract is also configured as a 1.0mg / mL standard solution. Take 0.5mL of each and place it in a 10mL volumetric flask, add 0.4mL of 5% sodium nitrite; after standing for 6 minutes, add 0.4mL of 10% aluminum nitrate; after standing for 6 minutes, add 4mL of 4% sodium hydroxide, add anhydrous ethanol to the scale line, and shake well; after standing for 15 minutes, measure the absorbance at a wavelength of 510nm. Draw a standard curve based on the absorbance of the gradient solution, and calculate the content of flavonoids in the sample using the standard curve.

[0057] 3. Experimental results and conclusions.

[0058] Table 1 Ethanol extract yields and flavonoid contents of eight kinds of Dendrobium officinale dry products

[0059]

[0060] Table 1 shows the yields of ethanol extracts and the flavonoid contents of the eight Dendrobium officinale varieties. While the yield of the ethanol extract of the Ziyu raw material was lower than that of Dendrobium officinale No. 2, Longzi No. 1, and Longzi No. 5, it was comparable to that of Dendrobium officinale No. 1 and significantly higher than that of Longzi No. 2, Longzi No. 4, and Wujincao. Furthermore, the Ziyu raw material had the highest flavonoid content, reaching 17.51%, significantly higher than the other seven Dendrobium officinale varieties.

[0061] Example 3

[0062] Qualitative analysis of flavonoids contained in purple jade.

[0063] 1. Reagents.

[0064] Apigenin, vetrin-2, rutin, quercetin-3-O-rutinoside-7-O-glucoside, methanol, purified water.

[0065] 2. Experimental Methods

[0066] Accurately weigh an appropriate amount of the ethanol extract of Ziyu described in Example 1 into a 1.5 mL EP tube and dissolve it in the corresponding volume of methanol to prepare a 2.0 mg / mL solution. Filter the filtrate through a 0.22 μm microporous membrane to obtain the test solution. Accurately weigh appropriate amounts of four standards (apigenin, vetamine-2, rutin, and quercetin-3-O-rutinoside) into a 1.5 mL EP tube and dissolve them in the corresponding volume of methanol to prepare a 2.0 mg / mL reference stock solution. Mix the solutions in a 1:1:1:1 ratio to obtain a mixed standard reference solution. LC-MS conditions: Kromasil 100-5C18 column (250 mm × 4.6 mm, 5 μm); column temperature 25°C, flow rate 1 mL / min; mobile phase water (A)-acetonitrile (B); elution gradient: 0.0–30.0 min, 97%–50% A, 5%–95% B; 30.0–31.0 min, 50%–0% A, 50%–100% B; 31.0–35.0 min, 0% A, 100% B; injection volume 20 μL. The mass detector was set to ESI negative ion mode.

[0067] According to the same method, the seven species of Dendrobium officinale except Ziyuzhi in Example 2 were also qualitatively analyzed for the flavonoid compounds contained therein.

[0068] 3. Experimental results and conclusions.

[0069] Table 2 Analysis of specific flavonoid components in eight species of Dendrobium dentata

[0070]

[0071] The present invention uses LC-MS to qualitatively analyze the chemical components of flavonoids in purple jade, and obtains the mass spectra of vecinin-2 and rutin contained in purple jade. Figure 2 By comparing the retention times with those of four flavonoid standards, it can be determined that Ziyu contains vecinin-2 and rutin.

[0072] The results of flavonoid component analysis of eight kinds of Dendrobium officinale ethanol extracts by LC-MS are shown in Table 2. According to the determination results shown in Table 2, Dendrobium officinale No. 1 and Dendrobium officinale No. 2 contain apigenin, and except Dendrobium officinale No. 1, all the Dendrobium officinale ethanol extracts contain rutin. Only Ziyu contains vecinin-2, and quercetin-3-O-rutinoside-7-O-glucoside was not detected in any of the eight Dendrobium officinale extracts.

[0073] Example 3

[0074] Total antioxidant capacity determination.

[0075] 1. Reagents.

[0076] Total antioxidant capacity test kit (ABTS method), rutin, vetamine-2, L-ascorbic acid, and purified water.

[0077] 2. Experimental method.

[0078] The water-soluble vitamin E standard (Trolox, 6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid) was diluted to a series of gradient concentrations (0.1, 0.2, 0.4, 0.8, and 1.0 mM). The concentration of the ethanol extract of Ziyu was 0.25 mg / mL. The concentrations of rutin, vetamine-2, and the positive control were uniformly set at 25 μM. According to the kit instructions, samples, standards, application solutions, and working solutions were sequentially added to a 96-well plate. The absorbance of each well was measured, and the standard curve was used for calculations.

[0079] 3. Experimental results and conclusions.

[0080] Table 3 Total antioxidant capacity of jasmine extract, rutin and vecinin-2

[0081] Serial number Sample name Total antioxidant capacity (mM) 1 Positive control (L-ascorbic acid) 0.14 2 Purple Jade Extract 3.72 3 Rutin 1.27 4 Weicaining-2 1.03

[0082] The total antioxidant capacity of rutin, rutin and vetamine-2 was obtained as Figure 3 As shown in Table 3, this experiment evaluated the total antioxidant capacity of the ethanol extract of Ziyu, rutin, and Vecinin-2. The results showed that, under the experimental concentration conditions, the ethanol extract of Ziyu, rutin, and Vecinin-2 all exhibited significantly higher activity than the positive control, L-ascorbic acid. The ethanol extract of Ziyu exhibited excellent total antioxidant capacity, approaching 4 mM Trolox equivalent.

[0083] Example 4

[0084] Hydroxyl radical scavenging ability assay.

[0085] 1. Reagents.

[0086] Hydroxyl radical scavenging ability kit, rutin, vetamine-2, L-ascorbic acid, purified water.

[0087] 2. Experimental method.

[0088] The concentration of the ethanol extract of Ziyu was 0.25 mg / mL, and the concentrations of rutin, vetamine-2, and the positive control were uniformly set at 25 μM. According to the kit instructions, samples and reagents were added to a 96-well plate in sequence, and the absorbance of each well was measured. The absorbance was then calculated using the formula.

[0089] 3. Experimental results and conclusions.

[0090] Table 4 Hydroxyl radical scavenging ability of Ziyu extract, rutin and Weicaining-2

[0091] Serial number Sample name Hydroxyl radical scavenging ability (%) 1 Positive control (L-ascorbic acid) 7.91 2 Purple Jade Extract 24.61 3 Rutin 11.17 4 Weicaining-2 17.03

[0092] The hydroxyl radical scavenging ability of jasmine extract, rutin and vecinin-2 was obtained as Figure 4 As shown in Table 4, this experiment evaluated the hydroxyl radical scavenging activity of the ethanol extract of Ziyu, rutin, and Vecinin-2. The results showed that, under the experimental concentration conditions, the ethanol extract of Ziyu, rutin, and Vecinin-2 were significantly more active than the positive control, L-ascorbic acid. The ethanol extract of Ziyu exhibited an excellent hydroxyl radical scavenging activity, exceeding 20%, more than double that of the positive control.

[0093] Example 5

[0094] Superoxide anion inhibition ability assay.

[0095] 1. Reagents.

[0096] Tris-HCl, pyrogallol, rutin, vinblastine-2, L-ascorbic acid, purified water.

[0097] 2. Experimental method.

[0098] The concentration of the ethanol extract of Ziyu was 0.25 mg / mL, and the concentrations of rutin, vetamine-2, and the positive control were uniformly set at 25 μM. 30 μL of sample, 855 μL of Tris-HCl buffer (pH 8.2, 0.05 M), and 15 μL of pyrogallol solution (6 mM) were added to a 1.5 mL EP tube and thoroughly mixed. The absorbance was measured at 320 nm and recorded as A ...

[0099] 3. Experimental results and conclusions.

[0100] Table 5 Superoxide anion inhibition ability of Ziyu extract, rutin and Weicaining-2

[0101] Serial number Sample name Superoxide anion inhibition ability (%) 1 Positive control (L-ascorbic acid) 49.03 2 Purple Jade Extract 49.99 3 Rutin 52.32 4 Weicaining-2 52.31

[0102] The superoxide anion inhibition ability of ziyu extract, rutin and vetamine-2 was obtained as Figure 5 As shown in Table 5, this study evaluated the superoxide anion scavenging abilities of the ethanol extract of Ziyu, rutin, and Vecinin-2. The results showed that, under the experimental concentrations, the activities of the ethanol extract of Ziyu, rutin, and Vecinin-2 were slightly higher than those of the positive control, L-ascorbic acid. The activities of rutin and Vecinin-2 were significantly higher.

[0103] Example 6

[0104] DPPH free radical scavenging ability determination.

[0105] 1. Reagents.

[0106] DPPH free radical scavenging ability kit, rutin, vetamine-2, L-ascorbic acid, purified water.

[0107] 2. Experimental method.

[0108] The concentration of the ethanol extract of Ziyu was 0.25 mg / mL, and the concentrations of rutin, vetamine-2, and the positive control were uniformly set at 25 μM. According to the kit instructions, sample, methanol, and working solution were added to a 96-well plate in sequence. The absorbance of each well was measured and calculated according to the formula.

[0109] 3. Experimental results and conclusions.

[0110] Table 6 DPPH free radical scavenging ability of Ziyu extract, rutin and Weicaining-2

[0111]

[0112]

[0113] The DPPH free radical scavenging ability of ziyu extract, rutin and vecinin-2 was obtained. Figure 6 As shown in Table 6, the DPPH scavenging ability of the ethanol extract of Ziyu, rutin, and Weicainin-2 was evaluated in this experiment. The results showed that at a concentration of 25 μM, rutin had a higher activity than the positive control L-ascorbic acid.

[0114] Example 7

[0115] SOD inhibitory ability determination.

[0116] 1. Reagents.

[0117] SOD kit, rutin, vetamine-2, L-ascorbic acid, PBS, and purified water.

[0118] 2. Experimental method.

[0119] For HaCaT cells, 4 × 10 4 Cells were plated in 6-well plates at a density of 100 μM / mL. When the cells reached a density of approximately 80%, they were washed twice with PBS and incubated with samples and L-ascorbic acid for 12 hours. The concentrations of rutin, vetamine-2, and L-ascorbic acid were adjusted to 0.125, 0.25, 0.5, 1, and 2 mg / mL, and the concentrations of rutin, vetamine-2, and L-ascorbic acid were adjusted to 3.125, 6.25, 12.5, 25, and 50 μM. Proteins were extracted using Western blotting and IP buffers, and the collected proteins were concentrated to a uniform concentration.

[0120] According to the kit instructions, working solution, diluent and application solution were added to the 96-well plate in sequence. After incubation at 37°C for 20 minutes, the absorbance of each well was measured and finally calculated according to the formula.

[0121] 3. Experimental results and conclusions.

[0122] Table 7 SOD scavenging ability of Ziyu extract, rutin and Weicaining-2

[0123]

[0124]

[0125] The SOD scavenging ability of Ziyu extract, rutin and Weicaining-2 was obtained as Figure 7 As shown in Table 7, this study evaluated the intracellular SOD inhibitory abilities of the ethanol extract of Ziyu, rutin, and Vecinin-2. The results showed that the ethanol extract of Ziyu exhibited the best free radical inhibitory activity. Compared to the blank control group without sample addition, it exhibited significant activity across the concentration range of 0.125 to 2 mg / mL, and was superior to the positive control L-ascorbic acid at the same concentration. Similarly, rutin demonstrated excellent inhibitory activity. However, at concentrations above 12.5 μM, the inhibition rate decreased slightly with increasing rutin concentration, likely due to the cytotoxicity of high rutin concentrations. Vecinin-2 also exhibited significant activity within the concentration range of 25 to 50 μM, with the activity exhibiting a dose-dependent pattern.

[0126] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creative work, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. A method for preparing a purple jade (Yunlin Garden Plant New Registration No. 20230032) extract, characterized in that: The following steps are involved: The purple jade is extracted with ethanol aqueous solution as solvent to obtain the purple jade extract.

2. The preparation method according to claim 1, characterized in that: The ethanol aqueous solution contains 80% to 95% ethanol by volume.

3. The preparation method according to claim 1, characterized in that: After the extraction, the filtrate is filtered, concentrated and dried.

4. The preparation method according to claim 1, characterized in that: Before the extraction, the purple jade is crushed, and the particle size of the crushed jade is 20 to 60 meshes.

5. The purple jade extract prepared by the preparation method according to any one of claims 1 to 4.

6. The purple jade extract according to claim 5, characterized in that The purple jade extract contains flavonoids; the flavonoids include rutin and vecinin-2.

7. Use of the purple jade extract prepared by the preparation method according to any one of claims 1 to 4 or the purple jade extract according to claim 5 or 6 in the preparation of antioxidant products.

8. Use of the purple jade extract prepared by the preparation method according to any one of claims 1 to 4 or the purple jade extract according to claim 5 or 6 in the preparation of antioxidant foods.

9. Use of the purple jade extract prepared by the preparation method according to any one of claims 1 to 4 or the purple jade extract according to claim 5 or 6 in the preparation of antioxidant drugs.

10. Use of the purple jade extract prepared by the preparation method according to any one of claims 1 to 4 or the purple jade extract according to claim 5 or 6 in the preparation of daily chemical products.

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