Penthorum chinense pursh compound with effects of dispelling effects of alcohol and protecting liver and preparation method of penthorum chinense pursh compound

Abstract

The invention relates to the technical field of traditional Chinese medicine extracts, and discloses a penthorum chinense pursh compound with hangover alleviating and liver protecting effects and a preparation method thereof.The preparation method comprises the following steps that 1, penthorum chinense pursh leaves and black tartary buckwheat bran are mixed, precooled and subjected to low-temperature superfine grinding; (2) adding compound enzymes (hemicellulase, beta-glucosidase and beta-galactosidase) into the raw materials subjected to superfine grinding, and carrying out enzymolysis treatment; (3) carrying out two times of down-flow three-stage extraction on the raw materials in the step (2); step (4): adding a gastrodia elata extract and a phlorizin extract; and (4) filtering by adopting a ceramic membrane. According to the preparation method provided by the invention, the extraction efficiency of key components of penthorum chinense pursh is improved, and the penthorum chinense pursh leaves, the black tartary buckwheat bran, the gastrodia elata and the phlorizin extract are compounded, so that the stability of the obtained compound is improved, the stability is improved, a good synergistic interaction effect is generated, and the compound shows a remarkable effect in the aspects of dispelling the effects of alcohol and protecting the liver.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine extracts, and in particular to a Herba Glechomae serratae complex with alcohol-relieving and liver-protecting effects and a preparation method thereof. Background Art

[0002] The statements in this section merely provide background information related to the present disclosure and may not constitute prior art.

[0003] Penthorum chinense Pursh, a traditional Miao medicine from the Gulin region, is rich in a variety of active ingredients. Flavonoids such as quercetin, pinosin, pinosin, apigenin, myricetin, and kaempferol, as well as organic acids such as gallic acid, brevicornic acid, and chebulic acid, are believed to be key components of Penthorum chinense's hepatoprotective activity. These active ingredients contribute to Penthorum chinense's outstanding hepatoprotective properties and are commonly used clinically to treat liver diseases such as alcoholic fatty liver disease and hepatitis. Therefore, it is crucial to extract and enrich these key components for further research and utilization.

[0004] The flavonoids and tannins in the herb are primarily present as various glycosides. Compared to flavonoid / tannin aglycones, these glycosides are relatively less water-soluble. Consequently, traditional extraction methods are unable to fully extract these glycosides, resulting in poor directional selectivity. Furthermore, the type, quantity, and location of the glycosides all affect their bioavailability. Traditional extraction methods produce herb leaf extracts with a distinct medicinal and green flavor. They also tend to precipitate during long-term storage, resulting in poor stability and hindering their application in food development. These issues have limited the research and utilization of herb and its key components. Therefore, there is an urgent need for methods that can fully extract the active ingredients in herb to improve its bioavailability and enhance the long-term storage stability of herb extracts. Summary of the Invention

[0005] The purpose of the present invention is to address the problem of low extraction efficiency of glycosides of flavonoids and tannins in the herb, and to provide a herb complex with alcohol-relieving and liver-protecting effects and a preparation method thereof. The herb is compounded with multiple substances to achieve a synergistic effect; its bioavailability is improved, and a large number of key components are enriched, and the storage stability is greatly enhanced.

[0006] The technical solutions of the present invention are as follows:

[0007] On the one hand, the present invention provides a Herba Gastrodiae with alcohol-relieving and liver-protecting effects, which comprises, by weight, 50-70 parts of Herba Gastrodiae leaves, 10-20 parts of black buckwheat bran, 5-25 parts of Gastrodia elata extract and 5-20 parts of phlorizin extract.

[0008] The compounded mixture achieved a synergistic effect, effectively reducing the blood alcohol concentration in mice in a drunken state. The synergistic effect was particularly strong when 64 parts of the herbaceous sedge leaf material, 18 parts of black buckwheat bran, 10 parts of Gastrodia elata extract, and 8 parts of phlorizin extract were added.

[0009] Preferably, the enzymatic hydrolysis extract of 58-64 parts of Herba Glechomae var. chinensis leaves and 15-18 parts of black buckwheat bran, the Gastrodia elata extract and 8-12 parts of phlorizin extract are prepared.

[0010] Another aspect of the present invention provides a method for preparing a Herba Glechomae Root complex having alcohol sobering and liver protecting effects, comprising the following steps:

[0011] Step (1): fully mix the raw material of yellow grass leaves and black buckwheat bran, pre-cool them, and grind them into ultrafine powder at low temperature.

[0012] Step (2): adding a complex enzyme to the crushed raw material for enzymatic hydrolysis and enzyme inactivation.

[0013] The complex enzyme is composed of hemicellulase, beta-glucosidase and beta-galactosidase in a ratio of 1:1:0 to 3:3:1.

[0014] Preferably, the complex enzyme is composed of hemicellulase, β-glucosidase and β-galactosidase in a ratio of 1:1:1 to 2:1:1.

[0015] Step (3): Divide the enzyme-inactivated Herba Lycopodii leaves into three equal parts, perform two downstream three-stage extractions, and collect the filtrates.

[0016] Step (4): add Gastrodia elata extract and phlorizin extract to the filtrate of step (3) and stir thoroughly.

[0017] Step (5): further filtering using a ceramic membrane.

[0018] According to a preferred embodiment, the precooling in step (1) is specifically to precool the crude raw material of the yellow grass leaves to a brittle state. More specifically, the precooling temperature is -40 to 0°C and the precooling time is 1 to 5 hours.

[0019] According to a preferred embodiment, the temperature of the low-temperature ultrafine grinding chamber in step (1) is 8 to 20° C., and the grinding time is 10 to 60 minutes.

[0020] According to a preferred embodiment, the enzymatic hydrolysis material-liquid ratio in step (2) is 1:2-5 (g / ml), the enzyme addition amount is 0.5%-2%, the enzymatic hydrolysis pH is 3-6, the enzymatic hydrolysis time is 1-3h, the enzymatic hydrolysis temperature is 30-60°C; the enzyme inactivation temperature is 90°C, and the time is 10min.

[0021] According to a preferred embodiment, in step (3), the material-liquid ratio is 1:10-50 (g / ml), the extraction temperature is 30-60° C., and the extraction time is 0.5-2.5 h.

[0022] According to a preferred embodiment, the ratio of the complex enzyme is preferably: hemicellulase, β-glucosidase, and β-galactosidase are combined in a ratio of 2:1:1.

[0023] Compared with the existing technology, the beneficial effects of the present invention are:

[0024] 1. A Herba Gastrodiae complex with alcohol-relieving and liver-protecting effects and a preparation method thereof. The complex provided by the present invention is based on Herba Gastrodiae leaves, supplemented with black buckwheat bran, Gastrodia elata and phlorizin. Black buckwheat bran is rich in bioflavonoids, chlorophyll, dietary fiber, and various trace elements; its characteristic flavonoid is rutin, which has antioxidant, liver-protecting, blood sugar-lowering, and blood pressure-lowering effects. Gastrodia elata belongs to the liver meridian and has the effects of calming liver yang, stopping wind and relieving spasms, and dispelling wind and unblocking meridians. Gastrodin is its most important active ingredient, which gives Gastrodia elata multiple effects such as protecting chemical liver damage, improving memory, protecting the nervous system, lowering blood lipids, and lowering blood pressure. Phlorizin belongs to the dihydrochalcone component of flavonoids and has multiple biological activities such as antioxidant, treating diabetes, and improving memory ability. The inventors of this application found that the combination of these four under certain conditions can not only improve the flavor and storage stability of the complex, but also give full play to the synergistic effect, so that the alcohol-relieving and liver-protecting effects can be enhanced;

[0025] 2. A Herba Adenophorae complex with alcohol sobering and liver protection effects and a preparation method thereof. The present invention utilizes a composite enzyme (hemicellulase, β-glucosidase, β-galactosidase) to enzymatically hydrolyze Herba Adenophorae leaves and black buckwheat bran, thereby destroying the cell wall structure of Herba Adenophorae leaves and black buckwheat bran, promoting the dissolution of a large amount of cell contents, and improving the water solubility and bioavailability of glycosides; especially when the ratio of hemicellulase, β-glucosidase, and β-galactosidase is 2:1:1;

[0026] 3. A herba scutellariae complex with alcohol sobering and liver-protecting properties and its preparation method. The present invention utilizes low-temperature ultrafine grinding to pre-treat herba scutellariae leaves and black buckwheat bran, ensuring thorough mixing and achieving uniform particle size and composition distribution after grinding. Low-temperature grinding reduces heat generated by mechanical action during the grinding process, thereby minimizing the loss of active and volatile components. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Schematic diagram of the turbidity of a Herba Camphoratae with alcohol sobering and liver protecting effects (from left to right are Examples 1 to 7);

[0028] Figure 2Schematic diagram of the turbidity of a Herba Glehniae complex with alcohol sobering and liver protecting effects (from left to right are comparative examples 1 to 6);

[0029] Figure 3 Schematic diagram of the changes in turbidity and centrifugal sedimentation rate of the Herba Camphoratae var. serratae complex with alcohol sobering and liver protecting effects prepared in Examples 1 to 7 and Comparative Examples 1 to 6;

[0030] Figure 4 HPLC chromatograms of gallic acid, quercetin, apigenin, kaempferol, and chondrin standards;

[0031] Figure 5 The GC-MS total ion chromatograms of the Herba Anemarrhenae complexes with alcohol detoxification and liver protection effects prepared in Examples 1 to 7 and Comparative Examples 1 to 6 correspond to the order in the upper right corner from top to bottom;

[0032] Figure 6 Schematic diagram of the open field test results of mice in the animal experiments of Examples 1 to 7 and Comparative Examples 1 to 6;

[0033] Figure 7 Schematic diagram of the test results of mouse liver function indicators in the animal experiments of Examples 1 to 7 and Comparative Examples 1 to 6;

[0034] Figure 8 Schematic diagram of the results of detecting mouse liver enzyme content in the animal experiments of Examples 1 to 7 and Comparative Examples 1 to 6;

[0035] Figure 9 Schematic diagram of the test results of blood ethanol concentration in mice in the animal experiments of Examples 1 to 7 and Comparative Examples 1 to 6. DETAILED DESCRIPTION

[0036] The specific embodiments listed in the present invention are only examples of the present invention, and the present invention is not limited to the specific embodiments described below. For those skilled in the art, any equivalent modifications and substitutions to the embodiments described below are also within the scope of the present invention. Therefore, equivalent transformations and modifications made without departing from the spirit and scope of the present invention should be included within the scope of the present invention. If specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. All reagents or instruments without the manufacturer specified are conventional products that can be purchased commercially. In order to better illustrate the present invention, numerous specific details are given in the specific embodiments below. Those skilled in the art should understand that the present invention can also be implemented without certain specific details. In other embodiments, methods, means, equipment and steps well known to those skilled in the art are not described in detail in order to highlight the main purpose of the present invention.

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by those skilled in the art. Unless otherwise specified, the units used in this specification are International Standard Units, and the numerical values ​​and numerical ranges appearing in this invention should be understood to include the inevitable systematic errors in industrial production.

[0038] The features and performance of the present invention are further described in detail below with reference to the embodiments.

[0039] Example 1 Preparation method of a Herba Glechomae serratae complex with alcohol sobering and liver protecting effects

[0040] The steps include:

[0041] Step (1): 58 parts of scutellaria baicalensis leaves and 15 parts of black buckwheat bran were mixed, pre-cooled, and then ultrafinely ground at low temperature. The pre-cooling temperature was -20°C for 2 hours. The ultrafine grinding chamber temperature was 12°C, and the grinding time was 15 minutes.

[0042] Step (2): Adding a composite enzyme to the crushed raw material for enzymatic hydrolysis and enzyme inactivation. The composite enzyme is composed of hemicellulase, β-glucosidase, and β-galactosidase in a ratio of 1:1:1; the enzymatic hydrolysis material-liquid ratio is 1:2 (g / ml), the enzyme addition amount is 0.8%, the enzymatic hydrolysis pH is 4.8, the enzymatic hydrolysis time is 2 hours, the enzymatic hydrolysis temperature is 40°C; the enzyme inactivation temperature is 90°C, and the time is 10 minutes.

[0043] Step (3): Divide the enzyme-inactivated Herba Chrysanthemi leaf material into three equal portions and perform two downstream three-stage extractions, collecting the filtrate. The material-liquid ratio is 1:20 (g / ml), the extraction temperature is 40°C, and the extraction time is 1.5 h.

[0044] Step (4): add 15 parts of Gastrodia elata extract and 12 parts of phlorizin extract to the extract in (3), and stir to mix.

[0045] Step (5): further filtering is performed using a ceramic membrane to obtain a Herba Camphoratae var. raphani complex having alcohol-relieving and liver-protecting effects.

[0046] Effect measurement methods

[0047] 1. Stability test

[0048] (1) The compound of the Herba Aspergiae glutinosae, which has the effect of sobering up and protecting the liver, was placed in a 37°C incubator for an accelerated storage test. Samples were taken at 0 days, 2 days, 4 days, 8 days, 16 days, 32 days, 64 days, and 128 days. After sampling, the OD values ​​were measured at a wavelength of 620 nm.

[0049] (2) The Ganhuangcao complex, which has the effects of detoxifying alcohol and protecting the liver, was placed in a 37°C incubator for an accelerated storage test. Samples were taken at 0 days, 2 days, 4 days, 8 days, 16 days, 32 days, 64 days, and 128 days. The samples were centrifuged at 4000 r / min for 10 minutes, and the supernatant was discarded. The precipitate was dried at 80°C to a constant weight, and the precipitate weight was recorded.

[0050] 2. Determination of total flavonoid content

[0051] Determination of total flavonoids content: NaNO2-Al(NO3)3-NaOH colorimetric method was used. 1 mL of the Herba Rutinae complex with the effect of sobering up and protecting the liver was transferred to a 25 mL colorimetric tube. 1 mL of 5% sodium nitrite solution was added first, shaken and allowed to stand for 5 minutes, 1 mL of 10% aluminum nitrate solution was added, shaken and allowed to stand for 5 minutes, 10 mL of 4% sodium hydroxide solution was added, and finally 70% ethanol was added, the volume was adjusted to 25 mL, mixed and allowed to stand for 15 minutes, and a color reaction was performed. The absorbance value was measured at a wavelength of 510 nm. A linear regression was performed on the absorbance (Y) of the rutin working solution and its concentration (X), and the regression equation was: Y = 11.909X + 0.0011, R 2 =0.998. The measured absorbance value was substituted into the regression equation to calculate the total flavonoid content.

[0052] 3. Determination of key ingredient content

[0053] High-performance liquid chromatography (HPLC) was used to determine the contents of quercetin, chondrostenone, kaempferol, apigenin, and gallic acid. 1 mL of each of the alcohol-detoxifying and liver-protecting ganhuangcao complexes was filtered through a 0.22 μm filter membrane and placed into a brown injection vial. The chromatographic column used was an InertSustain C18 (5 μm × 4.6 × 250 mm) mobile phase: 0.1% phosphoric acid (A) and acetonitrile (B); flow rate: 1 mL / min; elution gradient: 95% A to 75% A from 0 to 30 min; 75% A to 55% A from 30 to 55 min; 55% A to 95% A from 55 to 58 min; column temperature: 35°C; injection volume: 20 μL. Measurements were taken at wavelengths of 220 nm and 254 nm, and peak areas were recorded. Linear regression was performed using the absorbance (Y) of the working solutions of the five key components against their concentrations (X), yielding the regression equations. Substitute the measured peak areas into the regression equation and calculate their respective contents.

[0054] 4. Determination of volatile components

[0055] The composition and relative content differences of the volatile components in the examples and comparative examples were determined by headspace solid phase microextraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC-MS).

[0056] HS-SPME conditions: Accurately measure 5 mL of the test solution into a 20 mL headspace bottle, add 10 μL of 2-octanol solution with a mass concentration of 6.3 μg / mL, seal and place in a constant temperature water bath at 60°C, equilibrate for 5 minutes, extract and adsorb for 30 minutes (65-UM), and decompose at 250°C for 5 minutes. Repeat the experiment three times.

[0057] GC conditions: DB-5MS quartz capillary column (30 m × 0.25 mm × 0.25 μm); inlet temperature: 250°C; carrier gas: high-purity helium (≥99.999%); column flow rate: 1 mL / min (constant flow mode), splitless injection. Temperature program: initial temperature 50°C, hold for 5 min; increase at 4°C / min to 150°C, hold for 5 min; then increase at 10°C / min to 250°C, hold for 3 min; total run time: 48 min.

[0058] MS conditions: electron impact ion source (EI); transfer line temperature: 280°C, ion source: 230°C; ionization: 70 eV; quadrupole: 150°C; full scan; scan range: 40-600 m / z; interface: 250°C, column temperature: 40°C, injection port: 250°C.

[0059] 5. Animal Experiments

[0060] 1) Experimental Grouping: 154 SPF male balb / c mice were randomly divided into a model group, an embodiment group (seven groups), and a comparative example group (six groups), with 11 mice in each group.

[0061] 2) Administration: The Example and Comparative Example groups were given the corresponding test substances at 3 mL / kg BW for 7 consecutive days, while the control group was gavaged with an equal volume of normal saline. 30 minutes after the last gavage, each group was gavaged with 15 mL / kg of 56% ethanol.

[0062] 3) Detection indicators:

[0063] Observe the activity of mice: 8 mice were selected from each group. Unsteady crawling, dragging of the hind abdomen on the ground, and laziness with eyes closed were used as indicators of drunkenness. Free movement, flexible limbs, and mental recovery were used as indicators of sobriety. The drunkenness and soberness time of each group of mice were recorded, and the duration of drunkenness (min, the time from drunkenness to sobriety) was calculated.

[0064] Open field test: 30 minutes after the mice woke up from the alcohol, 8 mice from each group were taken to perform the open field test to assess the extent of changes in their mobility caused by alcohol consumption.

[0065] Serum liver function indicators: After the open field test, serum was collected from 8 mice in each group, and the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and total bilirubin (T-BIL) were determined by biochemical method.

[0066] Liver enzyme content: The livers of 8 mice in each group were collected and prepared into 10% liver homogenate. The levels of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) were determined by Elisa method.

[0067] Blood ethanol: 3 mice were selected from each group, and the ethanol content in the mice's blood was detected by biochemical method.

[0068] Example 2 Preparation method of a Herba Glechomae serratae complex with alcohol sobering and liver protecting effects

[0069] The steps include:

[0070] Step (1): 58 parts of scutellaria baicalensis leaves and 15 parts of black buckwheat bran were mixed, pre-cooled, and then subjected to low-temperature ultrafine grinding. The pre-cooling temperature was -20°C for 2 hours. The low-temperature ultrafine grinding chamber temperature was 12°C, and the grinding time was 25 minutes.

[0071] Step (2): Adding a composite enzyme to the crushed raw material for enzymatic hydrolysis and enzyme inactivation. The composite enzyme is composed of hemicellulase, β-glucosidase, and β-galactosidase in a ratio of 1:1:1; the enzymatic hydrolysis material-liquid ratio is 1:2 (g / ml), the enzyme addition amount is 0.8%, the enzymatic hydrolysis pH is 4.8, the enzymatic hydrolysis time is 2 hours, the enzymatic hydrolysis temperature is 40°C; the enzyme inactivation temperature is 90°C, and the time is 10 minutes.

[0072] Step (3): Divide the enzyme-inactivated Herba Chrysanthemi leaf material into three equal portions and perform two downstream three-stage extractions, collecting the filtrate. The material-liquid ratio is 1:20 (g / ml), the extraction temperature is 40°C, and the extraction time is 1.5 hours.

[0073] Step (4): add 15 parts of Gastrodia elata extract and 12 parts of phlorizin extract to the extract in (3), and stir to mix.

[0074] Step (5): further filtering is performed using a ceramic membrane to obtain a Herba Camphoratae var. raphani complex having alcohol-relieving and liver-protecting effects.

[0075] Effect measurement methods

[0076] Same as Example 1.

[0077] Example 3 Preparation method of a Herba Glechomae serratae complex with alcohol sobering and liver protecting effects

[0078] The steps include:

[0079] Step (1): 58 parts of scutellaria baicalensis leaves and 15 parts of black buckwheat bran were mixed, pre-cooled, and then ultrafinely ground at low temperature. The pre-cooling temperature was -20°C for 2 hours. The ultrafine grinding chamber temperature was 12°C, and the grinding time was 15 minutes.

[0080] Step (2): Adding a composite enzyme to the crushed raw material for enzymatic hydrolysis and enzyme inactivation. The composite enzyme is composed of hemicellulase, β-glucosidase, and β-galactosidase in a ratio of 2:1:0; the enzymatic hydrolysis material-liquid ratio is 1:2 (g / ml), the enzyme addition amount is 0.8%, the enzymatic hydrolysis pH is 4.8, the enzymatic hydrolysis time is 2 hours, the enzymatic hydrolysis temperature is 40°C; the enzyme inactivation temperature is 90°C, and the time is 10 minutes.

[0081] Step (3): Divide the enzyme-inactivated Herba Chrysanthemi leaf material into three equal portions and perform two downstream three-stage extractions, collecting the filtrate. The material-liquid ratio is 1:20 (g / ml), the extraction temperature is 40°C, and the extraction time is 1.5 h.

[0082] Step (4): add 15 parts of Gastrodia elata extract and 12 parts of phlorizin extract to the extract in (3), and stir to mix.

[0083] Step (5): further filtering is performed using a ceramic membrane to obtain a Herba Camphoratae var. raphani complex having alcohol-relieving and liver-protecting effects.

[0084] Effect measurement methods

[0085] Same as Example 1.

[0086] Example 4 Preparation method of a Herba Glechomae serratae complex with alcohol sobering and liver protecting effects

[0087] The steps include:

[0088] Step (1): 50 parts of scutellaria baicalensis leaves and 20 parts of black buckwheat bran were mixed, pre-cooled, and then subjected to low-temperature ultrafine grinding. The pre-cooling temperature was -20°C for 2 hours. The low-temperature ultrafine grinding chamber temperature was 12°C, and the grinding time was 15 minutes.

[0089] Step (2): Adding a composite enzyme to the crushed raw material for enzymatic hydrolysis and enzyme inactivation. The composite enzyme is composed of hemicellulase, β-glucosidase, and β-galactosidase in a ratio of 2:1:1; the enzymatic hydrolysis material-liquid ratio is 1:2 (g / ml), the enzyme addition amount is 0.8%, the enzymatic hydrolysis pH is 4.8, the enzymatic hydrolysis time is 2 hours, the enzymatic hydrolysis temperature is 40°C, and the enzyme inactivation temperature is 90°C and the time is 10 minutes.

[0090] Step (3): Divide the enzyme-inactivated Herba Chrysanthemi leaf material into three equal portions and perform two downstream three-stage extractions, collecting the filtrate. The material-liquid ratio is 1:20 (g / ml), the extraction temperature is 40°C, and the extraction time is 1.5 h.

[0091] Step (4): add 25 parts of Gastrodia elata extract and 5 parts of phlorizin extract to the extract in (3), and stir to mix.

[0092] Step (5): further filtering is performed using a ceramic membrane to obtain a Herba Camphoratae var. raphani complex having alcohol-relieving and liver-protecting effects.

[0093] Effect measurement methods

[0094] Same as Example 1.

[0095] Example 5 Preparation method of a Herba Glechomae serratae complex with alcohol sobering and liver protecting effects

[0096] The steps include:

[0097] Step (1): 58 parts of scutellaria baicalensis leaves and 15 parts of black buckwheat bran were mixed, pre-cooled, and then ultrafinely ground at low temperature. The pre-cooling temperature was -20°C for 2 hours. The ultrafine grinding chamber temperature was 12°C, and the grinding time was 15 minutes.

[0098] Step (2): Adding a composite enzyme to the crushed raw material for enzymatic hydrolysis and enzyme inactivation. The composite enzyme is composed of hemicellulase, β-glucosidase, and β-galactosidase in a ratio of 2:1:1; the enzymatic hydrolysis material-liquid ratio is 1:2 (g / ml), the enzyme addition amount is 0.8%, the enzymatic hydrolysis pH is 4.8, the enzymatic hydrolysis time is 2 hours, the enzymatic hydrolysis temperature is 40°C, and the enzyme inactivation temperature is 90°C and the time is 10 minutes.

[0099] Step (3): Divide the enzyme-inactivated Herba Chrysanthemi leaf material into three equal portions and perform two downstream three-stage extractions, collecting the filtrate. The material-liquid ratio is 1:20 (g / ml), the extraction temperature is 40°C, and the extraction time is 1.5 h.

[0100] Step (4): add 15 parts of Gastrodia elata extract and 12 parts of phlorizin extract to the extract in (3), and stir to mix.

[0101] Step (5): further filtering is performed using a ceramic membrane to obtain a Herba Camphoratae var. raphani complex having alcohol-relieving and liver-protecting effects.

[0102] Effect measurement methods

[0103] Same as Example 1.

[0104] Example 6 Preparation method of a Herba Glechomae serratae complex with alcohol sobering and liver protecting effects

[0105] The steps include:

[0106] Step (1): 64 parts of scutellaria baicalensis leaves and 18 parts of black buckwheat bran were mixed, pre-cooled, and then subjected to low-temperature ultrafine grinding. The pre-cooling temperature was -20°C for 2 hours. The low-temperature ultrafine grinding chamber temperature was 12°C, and the grinding time was 15 minutes.

[0107] Step (2): Adding a composite enzyme to the crushed raw material for enzymatic hydrolysis and enzyme inactivation. The composite enzyme is composed of hemicellulase, β-glucosidase, and β-galactosidase in a ratio of 2:1:1; the enzymatic hydrolysis material-liquid ratio is 1:2 (g / ml), the enzyme addition amount is 0.8%, the enzymatic hydrolysis pH is 4.8, the enzymatic hydrolysis time is 2 hours, the enzymatic hydrolysis temperature is 40°C, and the enzyme inactivation temperature is 90°C and the time is 10 minutes.

[0108] Step (3): Divide the enzyme-inactivated Herba Chrysanthemi leaf material into three equal portions and perform two downstream three-stage extractions, collecting the filtrate. The material-liquid ratio is 1:20 (g / ml), the extraction temperature is 40°C, and the extraction time is 1.5 h.

[0109] Step (4): add 10 parts of Gastrodia elata extract and 8 parts of phlorizin extract to the extract in (3), and stir to mix.

[0110] Step (5): further filtering is performed using a ceramic membrane to obtain a Herba Camphoratae var. raphani complex having alcohol-relieving and liver-protecting effects.

[0111] Effect measurement methods

[0112] Same as Example 1.

[0113] Example 7 Preparation Method of a Herba Glechomae Root Complex with Alcohol Relief and Liver Protection Efficacy

[0114] The steps include:

[0115] Step (1): 70 parts of scutellaria baicalensis leaves and 10 parts of black buckwheat bran were mixed, pre-cooled, and then subjected to low-temperature ultrafine grinding. The pre-cooling temperature was -20°C for 2 hours. The low-temperature ultrafine grinding chamber temperature was 12°C, and the grinding time was 15 minutes.

[0116] Step (2): Adding a composite enzyme to the crushed raw material for enzymatic hydrolysis and enzyme inactivation. The composite enzyme is composed of hemicellulase, β-glucosidase, and β-galactosidase in a ratio of 2:1:1; the enzymatic hydrolysis material-liquid ratio is 1:2 (g / ml), the enzyme addition amount is 0.8%, the enzymatic hydrolysis pH is 4.8, the enzymatic hydrolysis time is 2 hours, the enzymatic hydrolysis temperature is 40°C, and the enzyme inactivation temperature is 90°C and the time is 10 minutes.

[0117] Step (3): Divide the enzyme-inactivated Herba Chrysanthemi leaf material into three equal portions and perform two downstream three-stage extractions, collecting the filtrate. The material-liquid ratio is 1:20 (g / ml), the extraction temperature is 40°C, and the extraction time is 1.5 h.

[0118] Step (4): add 5 parts of Gastrodia elata extract and 15 parts of phlorizin extract to the extract in (3), and stir to mix.

[0119] Step (5): further filtering is performed using a ceramic membrane to obtain a Herba Camphoratae var. raphani complex having alcohol-relieving and liver-protecting effects.

[0120] Effect measurement methods

[0121] Same as Example 1.

[0122] Comparative Example 1: Preparation method of a Herba Glechomae serratae complex with alcohol-relieving and liver-protecting effects

[0123] This comparative example is a further improvement of Example 5. The difference from Example 5 is that the raw materials in this comparative example are not compounded with black buckwheat bran, Gastrodia elata extract and phlorizin extract.

[0124] The steps include:

[0125] Step (1): 58 parts of the raw material of the herb were pre-cooled and subjected to low-temperature ultrafine grinding. The pre-cooling temperature was -20°C for 2 hours. The low-temperature ultrafine grinding chamber temperature was 12°C and the grinding time was 15 minutes.

[0126] Step (2): Adding a composite enzyme to the crushed raw material for enzymatic hydrolysis and enzyme inactivation. The composite enzyme is composed of hemicellulase, β-glucosidase, and β-galactosidase in a ratio of 2:1:1; the enzymatic hydrolysis material-liquid ratio is 1:2 (g / ml), the enzyme addition amount is 0.8%, the enzymatic hydrolysis pH is 4.8, the enzymatic hydrolysis time is 2 hours, the enzymatic hydrolysis temperature is 40°C, and the enzyme inactivation temperature is 90°C and the time is 10 minutes.

[0127] Step (3): Divide the enzyme-inactivated Herba Chrysanthemi leaf material into three equal portions and perform two downstream three-stage extractions, collecting the filtrate. The material-liquid ratio is 1:20 (g / ml), the extraction temperature is 40°C, and the extraction time is 1.5 hours.

[0128] Step (4): further filtering using a ceramic membrane.

[0129] The effect measurement method is the same as in Example 1.

[0130] Comparative Example 2: Preparation method of a Herba Glechomae serratae complex with alcohol-relieving and liver-protecting effects

[0131] This comparative example is a further improvement of Example 5. The difference from Example 5 is that the raw materials in this comparative example do not contain black buckwheat bran.

[0132] The steps include:

[0133] Step (1): 58 parts of the raw material of the herb were pre-cooled and subjected to low-temperature ultrafine grinding. The pre-cooling temperature was -20°C for 2 hours. The low-temperature ultrafine grinding chamber temperature was 12°C and the grinding time was 15 minutes.

[0134] Step (2): Adding a composite enzyme to the crushed raw material for enzymatic hydrolysis and enzyme inactivation. The composite enzyme is composed of hemicellulase, β-glucosidase, and β-galactosidase in a ratio of 2:1:1; the enzymatic hydrolysis material-liquid ratio is 1:2 (g / ml), the enzyme addition amount is 0.8%, the enzymatic hydrolysis pH is 4.8, the enzymatic hydrolysis time is 2 hours, the enzymatic hydrolysis temperature is 40°C, and the enzyme inactivation temperature is 90°C and the time is 10 minutes.

[0135] Step (3): Divide the enzyme-inactivated Herba Chrysanthemi leaf material into three equal portions and perform two downstream three-stage extractions, collecting the filtrate. The material-liquid ratio is 1:20 (g / ml), the extraction temperature is 40°C, and the extraction time is 1.5 hours.

[0136] Step (4): add 15 parts of Gastrodia elata extract and 12 parts of phlorizin extract to the extract in (3), and stir to mix.

[0137] Step (5): further filtering is performed using a ceramic membrane to obtain a Herba Camphoratae var. raphani complex having alcohol-relieving and liver-protecting effects.

[0138] Effect measurement methods

[0139] Same as Example 1.

[0140] Comparative Example 3: Preparation method of a Herba Glechomae serratae complex with alcohol-relieving and liver-protecting effects

[0141] This comparative example is a further improvement of Example 5. The difference from Example 5 is that the raw materials in this comparative example are not compounded with Gastrodia elata extract and phlorizin extract.

[0142] The steps include:

[0143] Step (1): 58 parts of scutellaria baicalensis leaves and 15 parts of black buckwheat bran were mixed, pre-cooled, and then subjected to low-temperature ultrafine grinding. The pre-cooling temperature was -20°C for 2 hours. The low-temperature ultrafine grinding chamber temperature was 12°C, and the grinding time was 15 minutes.

[0144] Step (2): Adding a composite enzyme to the crushed raw material for enzymatic hydrolysis and enzyme inactivation. The composite enzyme is composed of hemicellulase, β-glucosidase, and β-galactosidase in a ratio of 2:1:1; the enzymatic hydrolysis material-liquid ratio is 1:2 (g / ml), the enzyme addition amount is 0.8%, the enzymatic hydrolysis pH is 4.8, the enzymatic hydrolysis time is 2 hours, the enzymatic hydrolysis temperature is 40°C, and the enzyme inactivation temperature is 90°C and the time is 10 minutes.

[0145] Step (3): Divide the enzyme-inactivated Herba Chrysanthemi leaf material into three equal portions and perform two downstream three-stage extractions, collecting the filtrate. The material-liquid ratio is 1:20 (g / ml), the extraction temperature is 40°C, and the extraction time is 1.5 hours.

[0146] Step (4): further filtering with a ceramic membrane to obtain a Herba Camphoratae var. raphanifoliae complex having the effects of sobering up and protecting the liver.

[0147] Effect measurement methods

[0148] Same as Example 1.

[0149] Comparative Example 4: Preparation method of a Herba Glechomae serratae complex with alcohol-relieving and liver-protecting effects

[0150] This comparative example is a further improvement of Examples 1-3 and Example 5. Unlike the aforementioned examples, the raw materials in this comparative example are not pre-cooled and ultrafinely crushed at low temperature, nor are they subjected to complex enzyme hydrolysis.

[0151] The steps include:

[0152] Step (1): 58 parts of scutellaria baicalensis leaves and 15 parts of black buckwheat bran were mixed. The mixed raw materials were divided into three equal parts and subjected to two downstream three-stage extractions, and the filtrate was collected. The extraction material-liquid ratio was 1:20 (g / ml), the extraction temperature was 40°C, and the extraction time was 1.5 hours.

[0153] Step (2): add 15 parts of Gastrodia elata extract and 12 parts of phlorizin extract to the extract in step (1), and stir to mix.

[0154] Step (3): further filtering using a ceramic membrane.

[0155] Effect measurement methods

[0156] Same as Example 1.

[0157] Comparative Example 5: Preparation method of a Herba Glechomae serratae complex with alcohol-relieving and liver-protecting effects

[0158] This comparative example is a further improvement of comparative example 3. Unlike the above examples, the raw materials in this comparative example are not pre-cooled and ultra-finely crushed at low temperature.

[0159] The steps include:

[0160] Step (1): 58 parts of yellow sedge leaves and 15 parts of black buckwheat bran are mixed.

[0161] Step (2): Adding a composite enzyme to the raw material for enzymatic hydrolysis and enzyme inactivation. The composite enzyme is composed of hemicellulase, β-glucosidase, and β-galactosidase in a ratio of 2:1:1; the enzymatic hydrolysis material-liquid ratio is 1:2 (g / ml), the enzyme addition amount is 0.8%, the enzymatic hydrolysis pH is 4.8, the enzymatic hydrolysis time is 2 hours, the enzymatic hydrolysis temperature is 40°C; the enzyme inactivation temperature is 90°C, and the time is 10 minutes.

[0162] Step (3): Divide the enzyme-inactivated Herba Chrysanthemi leaf material into three equal portions and perform two downstream three-stage extractions, collecting the filtrate. The material-liquid ratio is 1:20 (g / ml), the extraction temperature is 40°C, and the extraction time is 1.5 hours.

[0163] Step (4): further filtering with a ceramic membrane to obtain a Herba Camphoratae var. raphanifoliae complex having the effects of sobering up and protecting the liver.

[0164] Effect measurement methods

[0165] Same as Example 1.

[0166] Comparative Example 6: Preparation method of a Herba Glechomae serratae complex with alcohol-relieving and liver-protecting effects

[0167] This comparative example is a further improvement of comparative example 3. Unlike the above examples, the raw materials in this comparative example are not subjected to complex enzyme hydrolysis.

[0168] The steps include:

[0169] Step (1): 58 parts of scutellaria baicalensis leaves and 15 parts of black buckwheat bran were mixed, pre-cooled, and then subjected to low-temperature ultrafine grinding. The pre-cooling temperature was -20°C for 2 hours. The low-temperature ultrafine grinding chamber temperature was 12°C, and the grinding time was 15 minutes.

[0170] Step (2): The crushed leaves of the Herba Camphoratae were divided into three equal parts and subjected to two downstream three-stage extractions, and the filtrates were collected. The material-liquid ratio was 1:20 (g / ml), the extraction temperature was 40°C, and the extraction time was 1.5 hours.

[0171] Step (3): further filtering is performed using a ceramic membrane to obtain a Herba Camphoratae var. raphani complex having alcohol-relieving and liver-protecting effects.

[0172] Effect measurement methods

[0173] Same as Example 1.

[0174] Statistics

[0175] Depend on Figures 1 to 3 As can be seen, the turbidity of Examples 1-7 and Comparative Examples 1-6 increased to varying degrees during the 128-day accelerated storage period. The overall trends were essentially the same: a rapid increase followed by a slowing of the rate of increase before gradually reaching stability. The overall trend of the centrifugal sedimentation rate during the accelerated storage process for Examples 1-7 was a rapid increase followed by a gradual stabilization. Furthermore, the centrifugal sedimentation rate over the 128-day period was significantly lower than that of Comparative Examples 1-6.

[0176] The HPLC chromatograms of gallic acid, quercetin, apigenin, kaempferol and chondrin standards are as follows: Figure 4 As shown, the GC-MS total ion current is as follows Figure 5The contents of total flavonoids and key components in the Herba Lycopodii leaf composites prepared in Examples 1 to 7 are shown in Tables 1 to 2:

[0177] Table 1 Total flavonoids content

[0178]

[0179]

[0180] Table 2 Key ingredient content

[0181]

[0182] As can be seen from Table 1-2, the contents of total flavonoids and key components in the Herba Lysimachiae leaf composites prepared in Examples 1 to 7 were significantly increased compared with those in Comparative Examples 1 to 6; among them, the total flavonoids content in Example 6 was the highest, which was 10.34 mg / mL, and the contents of gallic acid, kaempferol and chondrol were also the highest, which were 176.93, 127.57 and 13.07 μg / mL, respectively; while the contents of quercetin and apigenin were the highest in Example 7, which were 78.36 and 106.94 μg / mL, respectively.

[0183] The relative contents of volatile components in Examples 1 to 7 and Comparative Examples 1 to 6 are shown in Table 3:

[0184] Table 3 Relative content of volatile components

[0185]

[0186]

[0187] Note: “ / ” means not detected.

[0188] As shown in Table 3, (E)-2-nonenal and lauryl aldehyde were not detected in Examples 1-7, and 2-undecanone and geranylacetone were not detected in Examples 4-7. While the remaining volatile components were detected in each example, their relative contents were significantly reduced. This indicates that the medicinal and green flavors of the present invention can be significantly reduced.

[0189] The results of the animal experiments on Examples 1 to 7 and Comparative Examples 1 to 6 - the duration of drunkenness test are shown in Table 4:

[0190] Table 4 Animal experiment - drunkenness duration test results

[0191]

[0192] As shown in Table 4, compared with the model group and comparative examples 1 to 6, Examples 1 to 5 can delay the time to intoxication after oral administration of ethanol in mice to a certain extent and reduce the duration of intoxication behavior. In contrast, Examples 6 and 7 have similar effects, with no intoxication behavior in the mice.

[0193] Depend on Figure 6 As can be seen, the mice treated with Examples 1-7 showed significantly higher total distance traveled, number of squares crossed, and number of central squares crossed than the model group and Comparative Examples 1-6, while the time spent in the central square was significantly lower than the model group and Comparative Examples 1-6. Example 7 effectively increased the number of squares crossed by mice and reduced the time spent in the central square, while Example 6 was even more effective in increasing the total distance traveled by mice and the number of central squares crossed. This demonstrates that the Ganhuangcao complex of this application has a good hangover-relief effect.

[0194] Depend on Figure 7 It can be seen that Examples 1 to 7 can all reduce the levels of AST, ALT, and T-BIL in the serum of mice after drunkenness. The levels of AST and T-BIL in Example 7 were the lowest, at 19.247±2.119 U / L and 5.653±1.048 μmol / L, respectively. The level of ALT in Example 6 was the lowest, at 6.371±2.257 U / L. AST and ALT are important indicators of liver function, and their increase usually means that liver cells are damaged. In addition, when liver cells are damaged, the liver's ability to process bilirubin decreases, resulting in the accumulation of bilirubin in the blood, and the T-BIL (total bilirubin) content will also increase.

[0195] Depend on Figure 8 As shown, the ADH and ALDH levels in the livers of mice treated with Examples 1 to 7 were significantly higher than those in the model group and Comparative Examples 1 to 6. The livers of mice treated with Example 6 had the highest ADH and ALDH levels, at 1.972±0.027 pg / mg and 1.899±0.076 ng / mg, respectively. Increased ADH and ALDH levels in the liver contribute to alcohol metabolism and detoxification.

[0196] Depend on Figure 9 It can be seen that Examples 1 to 7 can all effectively reduce the blood ethanol concentration of mice in a drunken state. The blood ethanol concentration of mice treated with Example 6 was the lowest, at 20.531±2.209 mmol / L.

[0197] In summary, to enhance the alcohol-detoxifying and liver-protecting effects of the Herba Adenophorae flavescentis complex, the conditions described in Example 6 are optimal. The complex comprises 64 parts Herba Adenophorae flavescentis raw material, 18 parts Black Buckwheat bran, 10 parts Gastrodia elata extract, and 8 parts Phlorizin extract. The low-temperature ultrafine grinding chamber temperature is 12°C, and the grinding time is 15 minutes. The hemicellulase, β-glucosidase, and β-galactosidase are combined in a ratio of 2:1:1.

[0198] The above-described embodiments merely represent specific implementation methods of the present application. While the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of protection of the present application. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the technical concept of the present application, and all such variations and improvements fall within the scope of protection of the present application.

Claims

1. A Herba Lysimachiae complex having the effects of sobering up and protecting the liver, characterized in that: The invention comprises, by weight, 50 to 70 parts of radix strychnifoliae leaves, 10 to 20 parts of black buckwheat bran, 5 to 25 parts of gastrodia elata extract and 5 to 20 parts of phloridzin extract.

2. The Herba Lysimachiae complex having the effects of sobering up and protecting the liver according to claim 1, characterized in that: In parts by mass, the ingredients include 58-70 parts of Herba Lysimachiae leaves, 10-15 parts of black buckwheat bran, 5-15 parts of Gastrodia elata extract and 8-15 parts of phloridzin extract.

3. A method for preparing a compound of Herba Chrysanthemi Indici with alcohol sobering and liver protecting effects, characterized in that: The steps include: Step (1): fully mixing the raw material of yellow grass leaves and black buckwheat bran, precooling, and ultrafine grinding at low temperature; Step (2): adding a complex enzyme to the crushed raw material for enzymolysis and enzyme inactivation; The complex enzyme is composed of hemicellulase, β-glucosidase and β-galactosidase in a ratio of 1:1:0 to 3:3:1; Step (3): Divide the enzyme-inactivated yellow grass leaf material into three equal portions, perform two downstream three-stage extractions, and collect the filtrate; Step (4): Add the Gastrodia elata extract and the phlorizin extract to the filtrate of step (3) and stir thoroughly. Step (5): further filtering using a ceramic membrane.

4. The method for preparing the Herba Chrysanthemi Indici with alcohol sobering and liver protecting effects according to claim 3, characterized in that: The precooling temperature in step (1) is -40 to 0°C, and the precooling time is 1 to 5 hours.

5. The method for preparing the Herba Chrysanthemi Indici with alcohol sobering and liver protecting effects according to claim 3, characterized in that: The temperature of the low-temperature ultrafine grinding chamber in the step (1) is 8 to 20° C., and the grinding time is 10 to 60 minutes.

6. The method for preparing the Herba Chrysanthemi Indici with alcohol sobering and liver protecting effects according to claim 3, characterized in that: The enzymatic hydrolysis material-liquid ratio in the step (2) is 1:2-5 (g / ml), the enzyme addition amount is 0.5%-2%, the enzymatic hydrolysis pH is 3-6, the enzymatic hydrolysis time is 1-3h, the enzymatic hydrolysis temperature is 30-60°C; the enzyme inactivation temperature is 90°C, and the time is 10min.

7. The method for preparing the Herba Chrysanthemi Indici with alcohol sobering and liver protecting effects according to claim 3, characterized in that: In the step (3), the material-liquid ratio is 1:10-50 (g / ml), the extraction temperature is 30-60° C., and the extraction time is 0.5-2.5 h.

8. The method for preparing the Herba Chrysanthemi Indici with alcohol sobering and liver protecting effects according to claim 1, characterized in that: In step (2), the ratio of the complex enzyme is: hemicellulase, β-glucosidase, and β-galactosidase are compounded in a ratio of 2:1:1.

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