Composition capable of inhibiting alpha-amylase and resisting oxidation and preparation method thereof

Abstract

The invention discloses a composition capable of inhibiting alpha-amylase and resisting oxidation and a preparation method thereof, and belongs to the field of traditional Chinese medicine extraction. The composition is prepared from the following components in percentage by mass: 20 to 30 percent of caulis polygoni multiflori, 20 to 40 percent of flos rosae rugosae, 10 to 30 percent of fructus amomi and 20 to 30 percent of rhizoma anemarrhenae. The preparation method comprises the following steps: crushing the medicinal materials, sieving with a 40-mesh sieve, adding 30-70% ethanol which is 10-20 times of the medicinal materials, carrying out 400-500W ultrasonic extraction for 2-3 times, and carrying out reduced pressure vacuum concentration and drying on the filtrate at 40-50 DEG C, thereby obtaining the traditional Chinese medicine composition. The alpha-amylase inhibition rate of the composition is 89.86%-96.07%, the ORAC value is 103238.56-184168.52 [mu] mol TE / L, the safety is high, the side effect is small, the process is convenient and fast, the composition can be used for assisting regulation and control of blood sugar, and the problems that existing chemical drugs are large in side effect, and traditional Chinese medicine combination effects are insufficient are solved.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine extraction technology, and specifically proposes a composition for inhibiting α-amylase and antioxidant and its preparation method. Background Technology

[0002] Diabetes mellitus is a prevalent endocrine and metabolic disorder worldwide. Its core pathological feature is an imbalance in glucose metabolism, which not only causes chronic damage to multiple organs but is also a significant risk factor for complications such as cardiovascular disease, neuropathy, and kidney disease, posing a serious threat to patients' health and quality of life.

[0003] As a major source of carbohydrates in the human diet, starch's digestion and metabolism are crucial in influencing postprandial blood glucose levels. Specifically, after entering the digestive tract, starch undergoes initial hydrolysis by salivary α-amylase, further degradation by pancreatic α-amylase, and finally conversion into glucose and absorption by brush border α-glucosidase in the small intestine. α-Amylase, as the core endogenous enzyme catalyzing the hydrolysis of α-1,4 glycosidic bonds in starch, breaks down starch into maltose, maltotriose, and a small amount of glucan, making it a key therapeutic target for regulating postprandial blood glucose elevation.

[0004] Currently, the main drugs used clinically to inhibit α-amylase include acarbose, miglitol, and voglibose. Although these drugs can inhibit enzyme activity by binding to the active site of α-amylase (hydrogen bonds or hydrophobic interactions), thereby slowing down starch digestion and glucose absorption and lowering postprandial blood glucose, long-term use can easily cause gastrointestinal adverse reactions such as bloating, diarrhea, and abdominal pain. Patients have poor tolerance to these drugs, which limits their clinical application.

[0005] Meanwhile, oxidative stress has been proven to be a significant contributing factor and aggravating factor in the occurrence and development of diabetes. When the body's oxidative stress level is abnormally elevated, excessive reactive oxygen species can damage the structure and function of pancreatic β-cells, leading to a decline in insulin secretion capacity; it can also induce insulin resistance, further exacerbating glucose metabolism disorders and forming a vicious cycle of "oxidative stress-glycemic imbalance." Currently used chemical antioxidants (such as butylated hydroxyanisole, butylated hydroxytoluene, tert-butylhydroquinone, propyl gallate, etc.) can scavenge free radicals and alleviate oxidative damage to some extent, but studies have shown that they have potential toxicity and adverse reactions, making them unsuitable for long-term oxidative stress intervention in diabetic patients.

[0006] Therefore, developing a traditional Chinese medicine composition that is highly safe, has few side effects, and possesses both α-amylase inhibitory and antioxidant activities has become a key need to address the pain points of existing technologies. Summary of the Invention

[0007] In view of this, and considering the shortcomings of existing α-amylase inhibitors such as large side effects, high potential risks of antioxidants, and the single efficacy and lack of synergistic combination of traditional Chinese medicine related products, the purpose of this invention is to provide a composition and preparation method with α-amylase inhibition and antioxidant properties. This composition uses natural traditional Chinese medicine as raw materials and achieves synergistic effects of components through specific ratios. While ensuring high safety, it simultaneously enhances α-amylase inhibitory activity and antioxidant activity, providing a new solution for the adjunctive intervention of diabetes.

[0008] The technical solution of this invention is achieved as follows: This invention provides a composition with α-amylase inhibition and antioxidant properties, wherein the composition is composed of four medicinal materials—Polygonum multiflorum vine, rose, Amomum villosum, and Anemarrhena asphodeloides—in a specific mass percentage of 100%, and the mass percentage range of each medicinal material is as follows: Polygonum multiflorum vine: 20%-30%; Roses: 20%-40%; Amomum villosum: 10%-30%; Anemarrhena asphodeloides: 20%-30%.

[0009] In some embodiments, to further improve efficacy stability, the raw materials of the composition may be any of the following preferred mass percentages: Preferred option 1: 20% Polygonum multiflorum vine, 20% rose, 30% Amomum villosum, and 30% Anemarrhena asphodeloides; Preferred Option 2: Polygonum multiflorum vine 25%, rose 25%, Amomum villosum 25%, Anemarrhena asphodeloides 25%; Preferred Option 3: Polygonum multiflorum vine 28%, rose 30%, Amomum villosum 20%, Anemarrhena asphodeloides 22%; Preferred option 4: 30% Polygonum multiflorum vine, 40% rose, 10% Amomum villosum, and 20% Anemarrhena asphodeloides.

[0010] The origins of each medicinal material are clearly identified to ensure the stability of the active ingredients. Polygonum multiflorum vine: The dried vine stem of Polygonum multiflorum Thunb., a plant in the Polygonaceae family. It is sweet and slightly bitter in nature, and has the effects of nourishing the heart and calming the mind, nourishing blood and promoting blood circulation, and relieving itching. Its chemical components mainly include flavonoids, stilbene glycosides, anthraquinones, and polysaccharides. In addition, Polygonum multiflorum vine also has antioxidant, hypoglycemic, immunomodulatory, and lipid-regulating effects. Rose: The dried flower buds of the rose, a plant in the Rosaceae family. Rich in various active ingredients, such as volatile oils, polysaccharides, polyphenols, and flavonoids, it also contains essential nutrients for the human body, including linoleic acid, vitamins, amino acids, dietary fiber, and trace elements. Roses possess multiple functional activities, including antibacterial, antioxidant, blood sugar-lowering, blood lipid-lowering, anti-tumor, sedative, and immune-regulating effects. As a special flower with both edible and medicinal value, it has broad prospects for development and utilization. Amomum villosum Lour., A. villosum Lour. var. xanthioides T.L. Wuet Senjen, or A. longiligulare T.L. Wu are dried, mature fruits of plants belonging to the genus Amomum villosum of the ginger family (Zingiberaceae). It is a plant that is both food and medicine. It is warm in nature, pungent in taste, and enters the spleen and stomach meridians. It has potential effects such as gastrointestinal protection, anti-inflammation, analgesia, antidiarrheal, antibacterial, regulation of flora, hypoglycemia, and anti-oxidation. Anemarrhena asphodeloides: The dried rhizome of Anemarrhena asphodeloides Bge., a plant of the Liliaceae family. It has a bitter and sweet taste and is cold in nature. Anemarrhena asphodeloides contains a variety of active ingredients, including steroidal saponins, flavonoids, polysaccharides, volatile oils, and alkaloids. It has significant pharmacological activities in anti-inflammatory, anti-tumor, hypoglycemic, lipid-lowering, and antioxidant properties.

[0011] In a second aspect, the present invention also provides a method for preparing the above-mentioned composition, the method comprising two core steps: extraction of medicinal materials and concentration and drying, the specific process of which is as follows: Extraction of medicinal materials: Weigh each medicinal material according to the above-mentioned mass percentages, and first pulverize them. After pulverization, pass them through a 40-mesh sieve to ensure the uniformity of the raw materials. Add 10-20 times the volume of ethanol solution with a concentration of 30%-70% to the pulverized mixed medicinal materials, and perform ultrasonic extraction at a power of 400-500W. The extraction is performed 2-3 times, and the extraction time is 40-60 minutes each time. After the extraction is completed, cool the extract to room temperature, and separate the residue and filtrate using conventional filtration methods (such as filter paper filtration or filter cloth filtration). Combine the filtrates from multiple extractions. Concentration and drying: The combined filtrate is placed in a vacuum concentration device and concentrated and dried under vacuum at a temperature of 40-50°C to remove ethanol solvent and water, finally obtaining a composition with α-amylase inhibition and antioxidant properties.

[0012] The core technical principle of this invention lies in the synergistic effect of multiple active ingredients from medicinal herbs: The stilbene glycosides and flavonoids in Polygonum multiflorum vine, together with the steroidal saponins and flavonoids in Anemarrhena asphodeloides, can synergistically enhance the inhibitory ability against α-amylase, and more efficiently block starch hydrolysis by competitively binding to the enzyme's active site. The polyphenols and flavonoids in roses, along with the volatile oils and polysaccharides in cardamom, can synergistically eliminate excess reactive oxygen species in the body and alleviate the damage of oxidative stress to pancreatic β cells. The active ingredients of the four medicinal herbs (flavonoids, saponins, polysaccharides, volatile oils, etc.) work synergistically to target the two core pathological processes of diabetes: "starch digestion disorder" and "oxidative stress," thus avoiding the problem of insufficient efficacy of a single ingredient or partial combination.

[0013] In the field of natural products, traditional Chinese medicine (TCM) exhibits unique advantages in blood sugar regulation and oxidative stress improvement due to its characteristics of "multiple components, multiple targets, and low toxicity and side effects." Existing research has found that single herbs or combinations of herbs such as Polygonum multiflorum vine, rose, Amomum villosum, and Anemarrhena asphodeloides possess certain hypoglycemic and antioxidant activities. For example, the stilbene glycosides and flavonoids in Polygonum multiflorum vine can regulate blood sugar metabolism; the polyphenols and flavonoids in rose can scavenge free radicals; the volatile oil components of Amomum villosum can help improve metabolic disorders; and the steroidal saponins in Anemarrhena asphodeloides have dual potential for hypoglycemic and antioxidant effects. However, to date, no research has combined these four herbs in a specific ratio, nor has any technical solution been found that, after synergistic effects, can simultaneously achieve "highly effective inhibition of α-amylase" and "strong antioxidant effects." Furthermore, there is a lack of reports on natural combinations targeting the dual pathological mechanisms of "starch digestion disorder + oxidative stress" in diabetes.

[0014] The present invention has the following advantages over the prior art: High safety and few side effects The raw materials of the composition of the present invention are all natural Chinese medicines that are both food and medicine or have a clear history of medicinal use. They do not contain chemically synthesized components, thus avoiding the gastrointestinal side effects of chemical α-amylase inhibitors and the potential toxicity risks of chemical antioxidants. They are suitable for long-term use and have higher patient tolerance.

[0015] Significant efficacy and strong synergistic effect Excellent α-amylase inhibitory activity: Through the synergistic effect of the four medicinal materials, the composition of the present invention can inhibit α-amylase by 89.86%-96.07%, which is significantly higher than the combination without any one medicinal material. It can more effectively slow down the starch digestion rate, reduce the amount of glucose absorbed after meals, and thus stabilize the postprandial blood glucose level. Outstanding antioxidant capacity: The antioxidant capacity (measured by ORAC value) of the composition of this invention can reach 103238.56-184168.52 μmolTE / L, which is much higher than the "flavorless combination". It can effectively remove reactive oxygen species in the body, protect the function of pancreatic β cells, improve insulin resistance, and assist in regulating blood glucose metabolism from the source. Dual efficacy achieved simultaneously: Unlike existing products that "single-inhibit α-amylase" or "single-antioxidant", the composition of this invention can act on the two key aspects of diabetes, namely "starch digestion disorder" and "oxidative stress", to achieve dual intervention of "lowering blood sugar and resisting damage", making it applicable to a wider range of scenarios.

[0016] The process is convenient and easy to industrialize. The preparation method of this invention only requires three core steps: "pulverization and sieving, ultrasonic extraction, and vacuum concentration and drying". It does not require complex active ingredient separation and purification processes, nor does it require the addition of additional auxiliary substances. Moreover, ultrasonic extraction has high efficiency, and the concentration and drying temperature is mild. The overall process is simple to operate, low in cost, and low in energy consumption, making it suitable for large-scale industrial production and conducive to industrialization and market promotion. Detailed Implementation

[0017] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0018] Example 1 Step 1: Medicinal herb extraction. Weigh out 20% Polygonum multiflorum vine, 20% rose, 30% Amomum villosum, and 30% Anemarrhena asphodeloides by weight percentage. Mix the above herbs and pulverize them. After pulverizing, pass the mixture through a 40-mesh sieve. Add 20 times the volume of 70% ethanol solution to the sieved mixture and place it in an ultrasonic extraction device. Extract the mixture three times at 500W power, with each extraction lasting 60 minutes. After extraction, allow the extract to cool naturally to room temperature. Filter the mixture with filter paper to separate the residue from the filtrate. Collect the filtrates from each extraction and combine them to obtain the total filtrate.

[0019] Step 2: Concentration and Drying. The combined total filtrate is transferred to a vacuum concentration apparatus and concentrated and dried under reduced pressure at 45°C until all ethanol solvent and water are removed, yielding a composition with α-amylase inhibition and antioxidant properties.

[0020] Example 2 Step 1: Extraction of medicinal materials. Weigh out 25% of Polygonum multiflorum vine, 25% of rose, 25% of Amomum villosum, and 25% of Anemarrhena asphodeloides by weight percentage, mix them, pulverize them and pass them through a 40-mesh sieve; add 20 times the volume of 35% ethanol solution to the mixed medicinal materials, and extract them by ultrasonic extraction 3 times at 500W power, 60 minutes each time; after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0021] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the target composition.

[0022] Example 3 Step 1: Extraction of medicinal materials. Weigh out 28% Polygonum multiflorum vine, 30% rose, 20% Amomum villosum, and 22% Anemarrhena asphodeloides by weight percentage, mix and grind them, and pass them through a 40-mesh sieve; add 10 times the volume of 50% ethanol solution to the mixed medicinal materials, and extract by ultrasonication 3 times at 500W power, 60 minutes each time; after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0023] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the target composition.

[0024] Example 4 Step 1: Extraction of medicinal materials. Weigh out 30% Polygonum multiflorum vine, 40% rose, 10% Amomum villosum, and 20% Anemarrhena asphodeloides by weight percentage, mix and grind them, and pass them through a 40-mesh sieve; add 10 times the volume of 35% ethanol solution to the mixed medicinal materials, and extract by ultrasonic extraction 3 times at 500W power, 60 minutes each time; after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0025] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the target composition.

[0026] Example 5 Step 1: Extraction of medicinal materials. Weigh out 22% Polygonum multiflorum vine, 35% rose, 18% Amomum villosum, and 25% Anemarrhena asphodeloides by weight percentage, mix and grind them, and pass them through a 40-mesh sieve; add 15 times the volume of 45% ethanol solution to the mixed medicinal materials, and extract twice by ultrasonication at 450W power, 50 minutes each time; after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0027] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 42°C to obtain the target composition.

[0028] Example 6 Step 1: Extraction of medicinal materials. Weigh out 29% Polygonum multiflorum vine, 22% rose, 29% Amomum villosum, and 20% Anemarrhena asphodeloides by weight percentage, mix and grind them, and then pass them through a 40-mesh sieve. Add 18 times the volume of 60% ethanol solution to the mixed medicinal materials, and extract by ultrasonication 3 times at 480W power, 45 minutes each time. After extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0029] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 48°C to obtain the target composition.

[0030] Comparative Example 1 Step 1: Extraction of medicinal materials. Weigh 40% rose petals, 30% cardamom, and 30% anemarrhena asphodeloides (excluding Polygonum multiflorum vine) by weight percentage, mix and grind them, and pass them through a 40-mesh sieve; add 10 times the volume of 50% ethanol solution to the mixed medicinal materials, and extract three times with ultrasonic power at 500W, 60 minutes each time; after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0031] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the composition.

[0032] Comparative Example 2 Step 1: Extraction of medicinal materials. Weigh out 30% Polygonum multiflorum vine, 50% Amomum villosum, and 20% Anemarrhena asphodeloides (excluding rose petals) by weight percentage, mix and grind them, and pass them through a 40-mesh sieve; add 10 times the volume of 70% ethanol solution to the mixed medicinal materials, and extract three times with ultrasonic power at 500W, 60 minutes each time; after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0033] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the composition.

[0034] Comparative Example 3 Step 1: Extraction of medicinal materials. Weigh out 25% Polygonum multiflorum vine, 35% rose petals, and 40% Anemarrhena asphodeloides (excluding Amomum villosum) by weight percentage, mix and grind them, and pass them through a 40-mesh sieve; add 10 times the volume of 40% ethanol solution to the mixed medicinal materials, and extract by ultrasonic extraction 3 times at 500W power, 60 minutes each time; after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0035] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the composition.

[0036] Comparative Example 4 Step 1: Extraction of medicinal materials. Weigh out 40% Polygonum multiflorum vine, 30% rose petals, and 30% Amomum villosum (excluding Anemarrhena asphodeloides) by weight percentage, mix and grind them, and pass them through a 40-mesh sieve; add 10 times the volume of 70% ethanol solution to the mixed medicinal materials, and extract three times with ultrasonic power at 500W, 60 minutes each time; after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0037] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the composition.

[0038] Comparative Example 5 Step 1: Extraction of medicinal materials. Weigh out 50% Polygonum multiflorum vine and 50% rose (excluding Amomum villosum and Anemarrhena asphodeloides) by weight percentage, mix and grind them, and pass them through a 40-mesh sieve; add 10 times the volume of 50% ethanol solution to the mixed medicinal materials, and extract by ultrasonication 3 times at 500W power, 60min each time; after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0039] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the composition.

[0040] Comparative Example 6 Step 1: Extraction of medicinal materials. Weigh 20% Amomum villosum and 80% Anemarrhena asphodeloides (excluding Polygonum multiflorum vine and rose petals) by weight percentage, mix and grind them, and pass them through a 40-mesh sieve; add 10 times the volume of 60% ethanol solution to the mixed medicinal materials, and extract three times with ultrasonic power at 500W, 60 minutes each time; after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0041] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the composition.

[0042] Comparative Example 7 Step 1: Extraction of medicinal materials. Weigh out 35% rose petals, 35% cardamom, and 30% anemarrhena asphodeloides by weight percentage (if Polygonum multiflorum vine is missing, adjust the ratio of rose petals and cardamom), mix and grind them, and then pass them through a 40-mesh sieve; add 20 times the volume of 35% ethanol solution to the mixed medicinal materials, and extract three times with ultrasonic power at 500W, 60 minutes each time (process is the same as in Example 2); after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0043] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the composition.

[0044] Comparative Example 8 Step 1: Extraction of medicinal materials. Weigh out 35% Polygonum multiflorum vine, 30% Amomum villosum, and 35% Anemarrhena asphodeloides by weight percentage (adjust the ratio of Polygonum multiflorum vine and Anemarrhena asphodeloides if rose is missing), mix and grind them, and then pass them through a 40-mesh sieve; add 20 times the volume of 35% ethanol solution to the mixed medicinal materials, and extract three times with ultrasonic power at 500W, 60 minutes each time (process is the same as in Example 2); after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0045] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the composition.

[0046] Comparative Example 9 Step 1: Extraction of medicinal materials. Weigh out 30% Polygonum multiflorum vine, 35% rose petals, and 35% Anemarrhena asphodeloides (adjust the ratio of Polygonum multiflorum vine and Anemarrhena asphodeloides if Amomum villosum is missing), mix and pulverize them, and then pass them through a 40-mesh sieve; add 20 times the volume of 35% ethanol solution to the mixed medicinal materials, and extract them by ultrasonic extraction 3 times at 500W power, 60 minutes each time (process is the same as in Example 2); after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0047] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the composition.

[0048] Comparative Example 10 Step 1: Extraction of medicinal materials. Weigh out 35% Polygonum multiflorum vine, 35% rose petals, and 30% Amomum villosum (adjust the ratio of Polygonum multiflorum vine and rose petals if Anemarrhena asphodeloides is missing), mix and pulverize them, and then pass them through a 40-mesh sieve; add 20 times the volume of 35% ethanol solution to the mixed medicinal materials, and extract them by ultrasonic extraction 3 times at 500W power, 60 minutes each time (process is the same as in Example 2); after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0049] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the composition.

[0050] Comparative Example 11 Step 1: Extraction of medicinal materials. Weigh 100% Polygonum multiflorum vine (high dose of single medicinal material), crush it and pass it through a 40-mesh sieve; add 20 times the volume of 35% ethanol solution to Polygonum multiflorum vine, and extract it by ultrasonic extraction 3 times at 500W power, 60min each time (process is the same as in Example 2); after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0051] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the composition.

[0052] Comparative Example 12 Step 1: Extraction of medicinal materials. Weigh 100% Anemarrhena asphodeloides (high dose of single herb), pulverize and pass through a 40-mesh sieve; add 20 times the volume of 35% ethanol solution to Anemarrhena asphodeloides, and extract by ultrasonication 3 times at 500W power, 60min each time (process is the same as in Example 2); after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0053] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the composition.

[0054] Comparative Example 13 Step 1: Extraction of medicinal materials. Weigh out 25% of Polygonum multiflorum vine, 25% of rose, 25% of Amomum villosum, and 25% of Anemarrhena asphodeloides (same composition ratio as in Example 2), mix and pulverize them, and then pass them through a 40-mesh sieve; add 20 times the volume of 35% ethanol solution to the mixed medicinal materials, place them in a reflux extraction device, and reflux extract three times at 80°C, 60 minutes each time (replacing ultrasonic extraction with reflux extraction); after extraction, cool to room temperature, filter with filter paper, and combine the filtrates.

[0055] Step 2: Concentration and drying. The filtrate was concentrated and dried under reduced pressure at 45°C to obtain the composition.

[0056] Performance verification: 1. Assay for α-amylase inhibitory activity 1.1 Preparation of reagents and instruments: Using corn starch as a substrate, α-amylase was dissolved in pH 6.8 phosphate buffer and stored at -20℃; 1g of corn starch was weighed and dispersed in 50mL of pH 6.8 phosphate buffer, mixed well and then placed in a 73℃ water bath for 10min for gelatinization. During the gelatinization process, the mixture was continuously stirred with a magnetic stir bar to obtain a starch gelatinized solution; a fluorescent microplate reader and a 96-well plate were prepared.

[0057] 1.2 Measurement Procedure: Take 20 μL of the composition samples from Examples 1-6 and Comparative Examples 1-13, and 20 μL of pH 6.8 phosphate buffer (blank group), and add them sequentially to the corresponding wells of a 96-well plate; add 20 μL of α-amylase solution to each well, place the 96-well plate in a fluorescence microplate reader, and preheat at 37°C for 15 min; after preheating, add 60 μL of starch gelatinization solution to each well, return the plate to the microplate reader, and automatically shake to mix. Set the measurement wavelength to 660 nm, and record the absorbance using the microplate reader's software. The initial absorbance is recorded as f1, and the absorbance is measured every 2 min thereafter, for a total of 61 points (absorbance is recorded as f1, f2...f61).

[0058] 1.3 Calculation method: The area under the decay curve (AUC) is calculated according to the formula AUC=2×(f2+f3+...+f60)+f1+f61; then the α-amylase inhibition rate is calculated according to the formula Inhibition%=(AUC sample-AUC blank) / AUC sample×100%.

[0059] 2. ORAC antioxidant capacity determination 2.1 Preparation of reagents and instruments: Azo compound AAPH was used as the source of peroxy free radicals, sodium fluorescein was used as a fluorescent indicator, and the vitamin E water-soluble analog Trolox was used as a quantitative standard; a fluorescent microplate reader and a 96-well plate were prepared, and the fluorescence intensity detection mode was set (excitation wavelength 485nm, emission wavelength 528nm, sensitivity 80, light source is halogen lamp).

[0060] 2.2 Measurement Procedure: Take 20 μL of the composition samples from Examples 1-6 and Comparative Examples 1-13, 20 μL of blank solution (without antioxidant), and 20 μL of Trolox standard solutions of different concentrations, and add them sequentially to the corresponding wells of a 96-well plate; add 160 μL of fluorescein solution to each well, place the 96-well plate in a microplate reader, and incubate at 37°C for 30 min; after incubation, quickly add 20 μL of LAPH solution to each well, shake well, and immediately start measuring the fluorescence intensity. The initial fluorescence intensity is recorded as f0, and then measured every 2 min for 120 min (a total of 61 measurement points, with fluorescence intensities recorded as f0, f1...f61 sequentially).

[0061] 2.3 Calculation method: The area under the fluorescence decay curve (AUC) was calculated according to the formula AUC=2×(f1 / f0+...+f60 / f0+f61 / f0)-f61 / f0+1; the net area (NetAUC) was obtained by subtracting the AUC of the blank group; a standard curve was plotted with the content of Trolox standard solution as the ordinate and the corresponding NetAUC as the abscissa, and the regression equation was obtained. The NetAUC of the sample was substituted into the equation to calculate the ORAC value of the sample. The result is expressed as μmolTrolox equivalent / L (μmolTE / L).

[0062] The performance results are shown in the table below:

[0063] As can be seen from the data in the table above, the inhibition rates of α-amylase in Examples 1-4 were all at a high level of 89.86%-96.07%, and the ORAC values ​​of antioxidant capacity were all ≥103238.56μmolTE / L. Among them, Example 2 (four herbs in equal proportions) had the highest inhibition rate (96.07%), and Example 4 (40% rose and 10% cardamom) had the highest ORAC value (184168.52μmolTE / L). This proves that within the ratio range of "20%-30% Polygonum multiflorum, 20%-40% rose, 10%-30% cardamom, and 20%-30% anemarrhena asphodeloides", the four herbs can achieve excellent dual effects simultaneously through the synergistic effect of active ingredients. Example 5 (22% Polygonum multiflorum vine, 35% rose, 18% Amomum villosum, 25% Anemarrhena asphodeloides, 15 times 45% ethanol, 450W sonication twice) showed an α-amylase inhibition rate of 92.15% and an ORAC value of 145689.33 μmol TE / L; Example 6 (29% Polygonum multiflorum vine, 22% rose, 29% Amomum villosum, 20% Anemarrhena asphodeloides, 18 times 60% ethanol, 480W sonication three times) showed an inhibition rate of 93.82% and an ORAC value of 162457.89 μmol TE / L. Both indicators were within the efficacy range of Examples 1-4 (inhibition rate 89.86%-96.07%, ORAC value 103238.56-184168.52 μmol TE / L), and showed no significant fluctuations. This result proves that regardless of whether the composition ratio is the "mid-range value" (such as 22% / 35% / 18% / 25% in Example 5) or the "near-boundary value" (such as 29% / 22% / 29% / 20% in Example 6), and regardless of whether the process parameters are "15 times ethanol, 450W power, 2 extractions" (Example 5) or "18 times ethanol, 480W power, 3 extractions" (Example 6), as long as the core conditions of "complete combination of four medicinal materials + 10-20 times 30%-70% ethanol + 400-500W ultrasonic extraction" defined by this invention are met, high dual efficacy can be stably achieved.

[0064] Comparative Examples 1-6, lacking 1-2 medicinal materials (Comparative Example 1 lacked Polygonum multiflorum vine, Comparative Example 2 lacked rose, Comparative Example 3 lacked Amomum villosum, Comparative Example 4 lacked Anemarrhena asphodeloides, Comparative Example 5 lacked both Amomum villosum and Anemarrhena asphodeloides, and Comparative Example 6 lacked Polygonum multiflorum vine and rose), showed a significant decrease in dual efficacy: the α-amylase inhibition rate was only 15.81%-56.24% (the lowest being Comparative Example 3 lacking Amomum villosum, only 15.81%), and the ORAC value was only 21152.41-40238.63 μmolTE / L (the highest being Comparative Example 1 lacking Polygonum multiflorum vine, only 40238.63 μmolTE / L). Both indicators were less than 50% of those in Examples 1-4. This indicates that the absence of any one of the four medicinal herbs will disrupt the synergistic mechanism of the active ingredients. For example, the absence of Amomum villosum directly leads to a sharp drop in the α-amylase inhibition rate, and the absence of Polygonum multiflorum vine and rose significantly weakens the antioxidant capacity. This proves that none of the four medicinal herbs can be missing, and there is no possibility that "the high efficacy can still be maintained after the absence of any of the herbs". Comparative Examples 7-10, based on the absence of any one of the medicinal materials, increased the proportions of the remaining three medicinal materials (e.g., Comparative Example 7, lacking Polygonum multiflorum vine, increased rose and Amomum villosum to 35%; Comparative Example 9, lacking Amomum villosum, increased Polygonum multiflorum vine and Anemarrhena asphodeloides to 30% / 35%), but the dual efficacy was still far lower than that of Example 2 (four herbs in equal proportions, inhibition rate 96.07%, ORAC value 153704.22 μmolTE / L): α-amylase inhibition rate was only 18.95%-58.72% (the highest being 58.72% in Comparative Example 7, less than 61% in Example 2), and ORAC value was only 29654.78-48652.17 μmolTE / L (the highest being 48652.17 μmolTE / L in Comparative Example 7, less than 32% in Example 2).

[0065] The results indicate that even if the proportion of the remaining medicinal materials is adjusted in an attempt to "make up for the loss of flavor", the synergistic effect of the four medicinal materials cannot be reconstructed. For example, in the control group 9 which lacked Amomum villosum, even if the proportions of Polygonum multiflorum and Anemarrhena asphodeloides were increased, the inhibition rate was still only 18.95%, which is close to that of the original control group 3 which lacked Amomum villosum (15.81%). This proves that the core role of Amomum villosum in α-amylase inhibition is irreplaceable.

[0066] Comparative Examples 11 (100% Polygonum multiflorum vine) and 12 (100% Anemarrhena asphodeloides) were extracted from "single medicinal materials at high doses". Their α-amylase inhibition rates were only 28.65%-33.27% and their ORAC values ​​were only 24315.89-28976.45 μmol TE / L, which were far lower than those of Example 2 (96.07% and 153704.22 μmol TE / L).

[0067] This data proves that even when using high doses of a single herb (Polygonum multiflorum vine or Anemarrhena asphodeloides, both of which have been reported to lower blood sugar / antioxidant effects on their own), its efficacy is far less than that of a combination of four herbs. The active ingredients of a single herb are limited and cannot simultaneously meet the dual mechanism requirements of "inhibiting α-amylase" and "antioxidation". In contrast, the flavonoids, saponins, volatile oils and other components of the four herbs can complement each other and work synergistically to achieve a synergistic effect.

[0068] Comparative Example 13 replaced the "500W ultrasonic extraction" of the present invention with "80℃ reflux extraction" (other conditions are the same as in Example 2), and its α-amylase inhibition rate decreased to 68.93% (a decrease of 28.25% compared to Example 2), and its ORAC value decreased to 79543.21 μmolTE / L (a decrease of 48.2% compared to Example 2).

[0069] The results indicate that ultrasonic extraction is not a "simple choice of conventional processes," but a necessary condition for achieving high efficacy. Ultrasonic extraction can release active ingredients (such as flavonoids and saponins) in medicinal materials more fully through cavitation effects, and low-temperature extraction (ultrasound does not require high temperatures) can reduce the destruction of active ingredients. In contrast, the high temperature (80°C) of reflux extraction may lead to the degradation of some antioxidant components, and the extraction efficiency is also lower than that of ultrasound.

[0070] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A composition having α-amylase inhibition and antioxidant properties, characterized in that, The composition comprises the following medicinal materials in the following weight percentages: Polygonum multiflorum vine 20%-30%, rose 20%-40%, Amomum villosum 10%-30%, and Anemarrhena asphodeloides 20%-30%.

2. The composition according to claim 1, characterized in that, The medicinal materials are expressed in the following percentages by weight: Polygonum multiflorum vine 20%, rose petals 20%, Amomum villosum 30%, and Anemarrhena asphodeloides 30%.

3. The composition according to claim 1, characterized in that, The medicinal materials are expressed in the following percentages by weight: Polygonum multiflorum vine 25%, rose 25%, Amomum villosum 25%, and Anemarrhena asphodeloides 25%.

4. The composition according to claim 1, characterized in that, The medicinal materials are expressed in the following percentages by weight: Polygonum multiflorum vine 30%, rose petals 40%, Amomum villosum 10%, and Anemarrhena asphodeloides 20%.

5. The composition according to claim 1, characterized in that, The Polygonum multiflorum vine is the dried stem of Polygonum multiflorum of the Polygonaceae family; the Amomum villosum is the dried mature fruit of Amomum villosum, Amomum tsao-ko, or Amomum hainanense of the Zingiberaceae family; and the Anemarrhena asphodeloides is the dried rhizome of Anemarrhena asphodeloides of the Liliaceae family.

6. A method for preparing the composition having α-amylase inhibition and antioxidant properties according to any one of claims 1-5, characterized in that, Includes the following steps: (1) Extraction of medicinal materials: After crushing the medicinal materials according to the mass percentage of claim 1 through a 40-mesh sieve, add 10-20 times the volume of ethanol solution with a concentration of 30%-70%, and extract by ultrasonic extraction 2-3 times at a power of 400-500W, each time for 40-60min. After extraction, cool to room temperature, filter, and combine the filtrates. (2) Concentration and drying: The filtrate obtained in step (1) is concentrated and dried under reduced pressure at a temperature of 40-50°C to obtain the composition.

7. The preparation method according to claim 6, characterized in that, The volume of the ethanol solution in step (1) is 20 times the mass of the medicinal material, the concentration is 70%, the ultrasonic power is 500W, and the extraction is performed 3 times, each time for 60 minutes; the concentration and drying temperature in step (2) is 45℃.

8. The preparation method according to claim 6, characterized in that, The volume of the ethanol solution in step (1) is 10 times the mass of the medicinal material, the concentration is 35%, the ultrasonic power is 500W, and the extraction is performed 3 times, each time for 60 minutes; the concentration and drying temperature in step (2) is 45℃.

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