Method for increasing extraction of effective components of astragalus by using sugar

Abstract

The present application provides a method for increasing the extraction of effective components from Astragalus membranaceus. The method comprises the following steps: (1) preparing a sugar solution and / or a sugar alcohol solution; (2) soaking Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution; and (3) heating the Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution to extract the Astragalus membranaceus, and then filtering to obtain an Astragalus membranaceus extract. The above method can significantly increase the extraction rate of effective components in Astragalus membranaceus, thereby helping to improve the utilization rate and efficacy of Astragalus membranaceus medicinal materials.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of traditional Chinese medicine extraction, and in particular to a method for increasing the extraction of effective components of Astragalus membranaceus by using sugar. BACKGROUND

[0002] Currently, how to efficiently extract effective components of traditional Chinese medicine and ensure the safety of the extract is an important content in the field of traditional Chinese medicine extraction research. The traditional method of taking traditional Chinese medicine is mostly water decoction, and clinical experience and modern research show that this method can better ensure the effectiveness and safety of the extract than organic solvent extraction. Therefore, in the new drug registration and application of traditional Chinese medicine released in 2020, if the application and traditional use are consistent, the clinical trial can be appropriately reduced or exempted, such as the "Provisions on Simplifying the Registration and Approval Management of Traditional Chinese Medicine Compound Preparations of Ancient Classical Prescriptions" issued in 2018, which clearly states that the development of compound preparations of ancient classical prescriptions can be exempted from clinical trials, including preparation technology and records in ancient medical books. Modern research has found that the ethanol extraction efficiency of most components of traditional Chinese medicine is much higher than that of water decoction, but the toxicity of the extract increases. The main reason for the difference between the two extraction methods is that traditional Chinese medicine contains a variety of effective components with a wide range of polarity, and ethanol extraction increases the extraction rate of weakly polar and moderately polar components, thereby causing the effective components in the alcohol extract to be different from those in the traditional clinical decoction, which may have some impact on the safety of the extract. In addition, the traditional extraction method of traditional Chinese medicine, which is mostly water decoction, has the problems of low extraction rate and incomplete extraction.

[0003] Therefore, how to improve the extraction rate of traditional Chinese medicine extract while adhering to the past is an important research direction in the field of traditional Chinese medicine extraction research. SUMMARY

[0004] Therefore, the present application provides an efficient extraction method for Astragalus membranaceus decoction pieces, which uses a certain concentration of sugar water and / or sugar alcohol water instead of water as the extraction solvent. Compared with the traditional decoction method, the extraction rate is significantly improved, thereby improving the utilization rate and efficacy of Astragalus membranaceus medicinal materials. The sugar used in this study is a common auxiliary material for traditional Chinese medicine or edible sugar, and therefore has the characteristics of safety and low price, and does not affect the safety of the extract.

[0005] Specifically, according to one aspect of the present application, a method for increasing the extraction of effective components of Astragalus membranaceus by using sugar is provided, which comprises the following steps:

[0006] (1) preparing a sugar solution and / or a sugar alcohol solution;

[0007] (2) soaking Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution; and

[0008] (3) heating the Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution to extract the Astragalus membranaceus, and filtering to obtain an Astragalus membranaceus extract.

[0009] According to another aspect of the present application, there is provided a method for increasing the extraction of effective components from Astragalus membranaceus using sugar, characterized in that the method comprises the following steps:

[0010] (1) preparing a sugar solution and / or a sugar alcohol solution;

[0011] (2) soaking the Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution;

[0012] (3) heating the Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution to extract the Astragalus membranaceus, and filtering to obtain a first Astragalus membranaceus extract;

[0013] (4) adding the sugar solution and / or the sugar alcohol solution to the Astragalus membranaceus, and heating to extract the Astragalus membranaceus to obtain a second Astragalus membranaceus extract; and

[0014] (5) combining the first Astragalus membranaceus extract and the second Astragalus membranaceus extract to obtain an Astragalus membranaceus extract (i.e. a total Astragalus membranaceus extract or a third Astragalus membranaceus extract).

[0015] Further, the sugar is selected from one or more of monosaccharides, disaccharides and trisaccharides.

[0016] Further, the sugar alcohol is selected from one or more of sorbitol, mannitol, erythritol, maltitol, lactitol and xylitol.

[0017] Further, the monosaccharide is selected from one or more of glucose, fructose, galactose, mannose, galactose, sorbose, rhamnose, ribose, xylose and deoxyribose.

[0018] Further, the disaccharide is selected from one or more of maltose, sucrose, lactose and trehalose.

[0019] Further, the monosaccharide is glucose and / or fructose.

[0020] Further, the disaccharide is selected from one or more of maltose, sucrose and trehalose.

[0021] Further, the trisaccharide is raffinose.

[0022] Further, the disaccharide is sucrose.

[0023] Further, in the sugar solution or the sugar alcohol solution, the concentration of the sugar or the sugar alcohol ranges from 0.1 to 30 g / 100 mL.

[0024] Further, the concentration of the sugar or the sugar alcohol ranges from 0.25 to 10 g / 100 mL.

[0025] Further, the concentration of the sugar or the sugar alcohol ranges from 1 to 10 g / 100 mL.

[0026] Further, the concentration of the sugar or the sugar alcohol ranges from 1 to 1.25 g / 100 mL or the concentration of the sugar or the sugar alcohol is about 10 g / 100 mL.

[0027] Further, the effective ingredient comprises saponins and / or flavonoids.

[0028] Further, the saponins are triterpene saponins.

[0029] Further, the saponins are astragaloside IV.

[0030] Further, the flavonoids are isoflavonoids.

[0031] Further, the flavonoids are selected from one or more of calycosin-7-glucoside, ononin, and calycosin.

[0032] Further, in step (2), the soaking is room temperature soaking.

[0033] Further, the step of soaking comprises soaking the Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution at room temperature for 10-120 min.

[0034] Further, the step of soaking comprises soaking the Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution at room temperature for 20-60 min.

[0035] Further, the step of soaking comprises soaking the Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution at room temperature for about 30 min.

[0036] Further, the weight-to-volume ratio of the Astragalus membranaceus to the sugar solution and / or the sugar alcohol solution is 1:1-1:50.

[0037] Further, the weight-to-volume ratio of the Astragalus membranaceus to the sugar solution and / or the sugar alcohol solution is 1:1-1:20.

[0038] Further, the weight-to-volume ratio of the Astragalus membranaceus to the sugar solution and / or the sugar alcohol solution is 1:6-1:7.

[0039] Further, in step (3), the heating extraction is heating reflux extraction.

[0040] Further, the heating time is 20 min to 60 min.

[0041] Further, the heating time is about 30 min.

[0042] According to another aspect of the present application, there is provided a use of the above-mentioned method in preparing an Astragalus extract or an Astragalus concentrate.

[0043] Further, the Astragalus concentrate is obtained by concentrating the Astragalus extract (i.e. the Astragalus total extract or the third Astragalus extract) obtained in step (5) under a water bath at 20-100°C.

[0044] Further, the concentrating in step (5) is completed under a water bath at 60-80°C.

[0045] Further, the concentrating is completed under a water bath at about 70°C.

[0046] According to another aspect of the present application, there is provided a use of the above-mentioned method in preparing a pharmaceutical composition containing Astragalus.

[0047] According to another aspect of the present application, there is provided a use of the above-mentioned method in preparing a pharmaceutical preparation, a functional food, a health food and other products containing Astragalus.

[0048] Advantages of the present application:

[0049] Astragalus is a commonly used traditional Chinese medicinal material, which is the root of Astragalus membranaceus Bge. var. mongholicus (Bge.) Hsiao. It is dug in spring and autumn, and then the soil, roots and root heads are removed, and it is dried to 60-70% of dryness, and then it is dried after being straightened and bundled. The main functions and indications are: tonifying Qi and consolidating the exterior, removing toxins and discharging pus, diuresis and promoting tissue regeneration. The main chemical components of Astragalus include polysaccharides, saponins, flavonoids and amino acids, etc. The pharmacological effects of Astragalus include improving immune function, enhancing antioxidant, anti-radiation and anti-cancer effects, protecting heart, brain, kidney and lung, protecting brain cells, improving memory, relaxing vascular smooth muscle, hormone-like effect, antibacterial and antiviral effect, reducing blood lipids, reducing blood sugar, reducing complications of diabetes, etc. We found that, compared with the traditional water decocting method, the method of the present application can sufficiently improve the extraction rate of effective components of Astragalus. DETAILED DESCRIPTION

[0050] The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, rather than all the embodiments of the present application. Based on the embodiments of the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0051] It should be noted that the embodiments in the present application and the features in the embodiments can be combined with each other without conflict. The present application will be described in detail below with reference to the embodiments.

[0052] The application will be described in further detail below with reference to specific embodiments, which are not to be understood as limiting the scope of the application as claimed.

[0053] As described in the background section, the alcohol extract is different from the effective component composition of the traditional clinically used decoction, so the safety of the alcohol extract can be affected to a certain extent, but the extraction efficiency of the traditional water decoction is not high. In order to solve the above problems, the application provides a method for increasing the extraction of effective components of Astragalus membranaceus by using sugar, which comprises the following steps:

[0054] (1) preparing a sugar solution and / or a sugar alcohol solution;

[0055] (2) soaking Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution; and

[0056] (3) heating the Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution to extract the Astragalus membranaceus, and obtaining an Astragalus membranaceus extract through filtration.

[0057] The sugar solution and / or the sugar alcohol solution can improve the extraction rate of the effective components in Astragalus membranaceus mainly in relation to two factors:

[0058] (1) whether the liquid can infiltrate the solid decoction piece, which is related to the surface tension thereof. The smaller the surface tension coefficient (30×10 -3 N / m or so), the liquid can almost infiltrate the solid; the surface tension coefficient of water is relatively large, and it can only infiltrate some solids. The surface tension coefficient of the sugar water solution is smaller than that of pure water, so the solution can better enter the plant cells and release the effective components, thereby improving the infiltration of the Astragalus membranaceus decoction piece and improving the extraction efficiency of the effective components;

[0059] (2) sugar can form hydrogen bonds and other forces with the active components in the Astragalus membranaceus decoction piece, thereby improving the extraction efficiency of the Astragalus membranaceus extract components;

[0060] Therefore, compared with the water decoction method, the method can sufficiently improve the extraction rate of the effective components of Astragalus membranaceus.

[0061] In actual work, the extraction times of the alcohol and / or sugar alcohol solution can be determined according to actual needs, which can be 1 time or several times, for example, 2-10 times.

[0062] According to another aspect of the application, a method for increasing the extraction of effective components of Astragalus membranaceus by using sugar is provided, which is characterized in that the method comprises the following steps:

[0063] (1) preparing a sugar solution and / or a sugar alcohol solution;

[0064] (2) soaking Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution;

[0065] (3) heating the Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution to extract the Astragalus membranaceus, and filtering to obtain a first Astragalus membranaceus extract;

[0066] (4) adding the sugar solution and / or the sugar alcohol solution to the Astragalus membranaceus, and heating to extract the Astragalus membranaceus to obtain a second Astragalus membranaceus extract; and

[0067] (5) combining the first Astragalus membranaceus extract and the second Astragalus membranaceus extract to obtain an Astragalus membranaceus extract (i.e., a total Astragalus membranaceus extract or a third Astragalus membranaceus extract).

[0068] The term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, a concentration of "about" 0.5 g crude drug / mL means a concentration from 0.475 g crude drug / mL to 0.525 g crude drug / mL, but also expressly includes a concentration of exactly 0.5 g crude drug / mL.

[0069] In a preferred embodiment, the sugar is selected from one or more of monosaccharides, disaccharides, and trisaccharides.

[0070] In a preferred embodiment, the sugar alcohol is selected from one or more of sorbitol, mannitol, erythritol, maltitol, lactitol, and xylitol. In actual applications, the above sugar alcohols can be replaced by other sugar alcohols in the prior art according to actual conditions.

[0071] In a preferred embodiment, the monosaccharide is selected from one or more of glucose, fructose, galactose, mannose, galactose, sorbose, rhamnose, ribose, xylose, and deoxyribose. In actual applications, the above monosaccharides can be replaced by other monosaccharides in the prior art according to actual conditions.

[0072] In a preferred embodiment, the disaccharide is selected from one or more of maltose, sucrose, lactose, and trehalose. In actual applications, the above disaccharides can be replaced by other disaccharides in the prior art according to actual conditions.

[0073] In a preferred embodiment, the monosaccharide is glucose and / or fructose.

[0074] In a preferred embodiment, the disaccharide is selected from one or more of maltose, sucrose, and trehalose.

[0075] In a preferred embodiment, the trisaccharide is raffinose.

[0076] In order to further improve the extraction rate of effective components in Astragalus membranaceus, in a preferred embodiment, the disaccharide is sucrose.

[0077] In a preferred embodiment, the concentration of the sugar or the sugar alcohol in the sugar solution or the sugar alcohol solution ranges from 0.1 to 30 g / 100 mL.

[0078] In a preferred embodiment, the concentration of the sugar or the sugar alcohol ranges from 0.25 to 10 g / 100 mL.

[0079] In a preferred embodiment, the concentration of the sugar or the sugar alcohol ranges from 1 to 10 g / 100 mL.

[0080] In order to further improve the extraction rate of the active ingredients in Radix Astragali, in a preferred embodiment, the concentration of the sugar or the sugar alcohol ranges from 1 to 1.25 g / 100 mL or the concentration of the sugar or the sugar alcohol is about 10 g / 100 mL.

[0081] The term "about" or "approximately" in relation to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, a concentration of "about" 10 g / 100 mL means a concentration from 9.5 g / 100 mL to 10.5 g / 100 mL, but also expressly includes a concentration of exactly 10 g / 100 mL.

[0082] In a preferred embodiment, the active ingredients comprise saponins and / or flavonoids.

[0083] In a preferred embodiment, the saponins are triterpene saponins. In practical applications, the saponins include other saponins that have been identified in Radix Astragali by the prior art in addition to triterpene saponins.

[0084] In a preferred embodiment, the saponins are astragaloside A.

[0085] In a preferred embodiment, the flavonoids are isoflavonoids.

[0086] In practical applications, the flavonoids include other flavonoids that have been identified in Radix Astragali by the prior art in addition to isoflavonoids.

[0087] In a preferred embodiment, the flavonoids are selected from one or more of calycosin-7-glucoside, formononetin, and calycosin.

[0088] In a preferred embodiment, in step (2), the soaking is room temperature soaking.

[0089] In a preferred embodiment, the step of soaking comprises soaking the Radix Astragali in the sugar solution and / or the sugar alcohol solution at room temperature for 10 to 120 min.

[0090] In a preferred embodiment, the step of soaking comprises soaking the Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution at room temperature for 20-60 min.

[0091] In a preferred embodiment, the step of soaking comprises soaking the Astragalus membranaceus in the sugar solution and / or the sugar alcohol solution at room temperature for about 30 min.

[0092] The term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, a time of "about" 30 min means a time from 28.5 min to 31.5 min, but also expressly includes a time of exactly 30 min.

[0093] In a preferred embodiment, the weight to volume ratio of the Astragalus membranaceus to the sugar solution and / or the sugar alcohol solution is 1:1-1:50.

[0094] In a preferred embodiment, the weight to volume ratio of the Astragalus membranaceus to the sugar solution and / or the sugar alcohol solution is 1:1-1:20.

[0095] In a preferred embodiment, the weight to volume ratio of the Astragalus membranaceus to the sugar solution and / or the sugar alcohol solution is 1:6-1:7.

[0096] In a preferred embodiment, in step (3), the heating extraction is a heating reflux extraction.

[0097] In a preferred embodiment, the heating is for a time of 20 min to 60 min.

[0098] In a preferred embodiment, the heating is for a time of about 30 min.

[0099] The term "about" or "approximately" in reference to a numerical value means ±5% of the numerical value, but expressly includes the exact numerical value. For example, a time of "about" 30 min means a time from 28.5 min to 31.5 min, but also expressly includes a time of exactly 30 min.

[0100] According to another aspect of the present application, there is provided a use of the Astragalus membranaceus extract or the Astragalus membranaceus concentrate prepared according to the above method.

[0101] In a preferred embodiment, the Astragalus membranaceus concentrate is obtained by concentrating the Astragalus membranaceus extract (i.e. the Astragalus membranaceus total extract or the third Astragalus membranaceus extract) obtained in step (5) under a water bath at 20°C to 100°C.

[0102] In a preferred embodiment, in step (5), the concentrating is done under a water bath at 60°C to 80°C.

[0103] In a preferred embodiment, the concentration is done in a water bath at about 70°C.

[0104] The term "about" or "approximately" in relation to a numerical value means ± 5% of the numerical value, but expressly includes the exact numerical value. For example, a temperature of "about" 70°C means a range of 66.5°C to 73.5°C, but also expressly includes a temperature of exactly 70°C.

[0105] According to another aspect of the present application, there is provided a use of the above-mentioned method for preparing a pharmaceutical composition containing Radix Astragali.

[0106] According to another aspect of the present application, there is provided a use of the above-mentioned method for preparing a pharmaceutical preparation, functional food, health food and other products containing Radix Astragali.

[0107] The functional food is a food that can convincingly prove to have benefits on one or more functions of the body, has sufficient nutritional effects to improve health conditions or can reduce the incidence of diseases.

[0108] The health food refers to a food that claims to have specific health functions or is intended for supplementing vitamins and minerals, i.e. is suitable for specific groups of people to eat, has the function of regulating the body, is not intended for the treatment of diseases, and does not cause any acute, subacute or chronic harm to the human body.

[0109] The other products refer to all products other than pharmaceutical preparations, functional food and health food that are included in the prior art, including liquid and solid forms, etc.

[0110] Example 1: Experiment of extracting raw Radix Astragali decoction pieces with different sugar solutions

[0111] 1.1 Preparation method of different sugar-Radix Astragali extraction solutions (two times of extraction with 1% sugar)

[0112] The same batch of Radix Astragali decoction (from Hunyuan, Shanxi) was taken as 5 samples (HQ, HQ+1% sucrose, HQ+1% glucose, HQ+1% fructose, HQ+1% trehalose), each weighing 50g. 7 times the amount of water (350ml water), 1% glucose aqueous solution (3.5g glucose was added to 350ml), 1% fructose aqueous solution (3.5g fructose was added to 350ml), 1% sucrose aqueous solution (3.5g sucrose was added to 350ml), 1% trehalose aqueous solution (3.5g trehalose was added to 350ml) were added respectively, soaked for 30min, refluxed for 30min, filtered with gauze while hot, and the residue was added with 6 times the amount of water (300ml water), 1% glucose aqueous solution (3.0g glucose was added to 300ml), 1% fructose aqueous solution (3.0g fructose was added to 300ml), 1% sucrose aqueous solution (3.0g sucrose was added to 300ml), 1% trehalose aqueous solution (3.0g trehalose was added to 300ml) respectively, refluxed for 20min, filtered while hot, and the two filtrates were combined to obtain different sugar-Astragalus membranaceus extract.

[0113] 1.2.1 Instruments and reagents

[0114] 1.2.1 Instruments and reagents

[0115] Agilent 1260 type high performance liquid chromatograph equipped with DAD detector (Agilent, USA); Agilent 1200 type high performance liquid chromatograph equipped with ELSD detector (Agilent, USA); DZKW-4 type electronic constant temperature water bath (Beijing Zhongxingweiye Instrument Co., Ltd.); DK-98-IIA electric heating constant temperature water bath (Tianjin Test Instrument Co., Ltd.); ME204 / 02 type electronic balance [Mettler-Toledo Instrument (Shanghai) Co., Ltd., 1 / 10 million balance]; KQ-250DE type numerical control ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); HC-3018 high speed centrifuge (Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.); solid phase extraction column UniEIut C18 EC column (1000mg / 6ml) (Huapukeli Beijing Technology Co., Ltd.).

[0116] Reference substance Astragaloside IV (batch number CHB170727), Calycosin-7-glucoside (batch number CHB161105), Maianthridin (batch number CHB150517), and Calycosin (batch number CHB161104) were purchased from Chengdu Keluoma Biological Technology Co., Ltd., with purity of HPLC≥98%; acetonitrile (chromatographic pure, Thermo Fisher Scientific), methanol (chromatographic pure, Thermo Fisher Scientific), formic acid (analytical pure, purchased from National Pharmaceutical Group Chemical Reagent Co., Ltd.); and water (Vahaha pure water).

[0117] 1.2.2 Preparation of test solution

[0118] Preparation of test solution of saponin: 5 ml of different sugar-Astragalus extract was taken, 20 ml of 60% methanol was added, and ultrasonic mixing was performed. Then, 10 ml of ammonia water was added, mixed, centrifuged at 3600 r / min for 10 min, and the supernatant was passed through a Waters UniELut C18 EC column (1000 mg / 6 ml) and eluted with 10 ml of pure water and then 5 ml of methanol. The methanol eluate was collected, centrifuged at 8000 r / min for 5 min, and the supernatant was passed through a 0.45 μm microporous filter to obtain the test solution.

[0119] Preparation of test solution of flavonoids: 300 μl of different sugar-Astragalus extract was taken, 700 μl of water was added, and the mixture was centrifuged at 12000 r / min for 5 min in a 1.5 ml centrifuge tube. The supernatant was passed through a 0.45 μm microporous filter to obtain the test solution.

[0120] 1.2.3 Preparation of control solution

[0121] Astragaloside, calycosin-7-glucoside, formononetin, and calycosin were accurately weighed, and methanol was added to prepare a 1 mg / ml stock solution. The stock solution was diluted to different concentrations to establish a working curve.

[0122] The linear equation of astragaloside was lnY = 1.3271 lnX + 9.9314, r = 1.0000; the linear equation of calycosin-7-glucoside was Y = 35129X + 14, r = 0.9998; the linear equation of formononetin was Y = 10024X + 33, r = 0.9998; and the linear equation of calycosin was Y = 4656X + 25, r = 0.9995.

[0123] 1.2.4 Content determination method

[0124] The content determination method of saponin was as follows: Agilent 1200 liquid chromatograph (equipped with an ELSD detector) was used for analysis, the chromatographic column was Thermo Acclaim 120 C18 (250 x 4.6 mm, 5 μm), the mobile phase was 0.1% formic acid water (A) and 0.1% formic acid acetonitrile (B), gradient elution was performed as follows: 0-5 min, 5%-10% B; 5-10 min, 10%-32% B; 10-30 min, 32%-45% B; 30-35 min, 45%-95% B; 35-40 min, 95%-20% B, the column temperature was room temperature, the flow rate was 1 ml / min, the drift tube temperature was 100°C, the carrier gas flow rate was 2.5 l / min, and the injection volume was 20 μl.

[0125] Flavonoid content determination method: Agilent 1260 type high performance liquid chromatograph (with DAD detector) was used for analysis, chromatographic column: Thermo Acclaim 120 C18 (250x4.6mm, 5um); mobile phase: 0.1% formic acid aqueous solution (A) and 0.1% formic acid acetonitrile (B), gradient elution: 0-8min, 5%-20%B; 8-15min, 20%-25%B; 15-20min, 25%B; 20-30min, 25%-40%B; 30-40min, 40%-60%B; flow rate 1ml / min; detection wavelength 260nm; column temperature 25℃; injection volume 10ul.

[0126] 1.3 Experimental results

[0127] The results of determination of the content of effective components in different sugar-Astragalus extract are shown in Table 1.

[0128] Table 1 Determination results of effective component content in different sugar-Astragalus extract

[0129]

[0130] *P<0.05 VS water

[0131] As can be seen from Table 1, sucrose, glucose, fructose and trehalose (1g / 100ml concentration) can increase the content of effective components in Astragalus to varying degrees, among which sucrose and trehalose have the most obvious effect, but using sucrose is more economical than using trehalose.

[0132] Example two different sugar solution extraction of raw Astragalus slices

[0133] 2.1 Preparation method of different sugar-Astragalus extract (different 1% sugar extraction once)

[0134] The same batch of raw Astragalus slices (from Shanxi Hunyuan) was taken in four portions (HQ, HQ+1% glucose, HQ+1% sucrose, HQ+1% maltose), each 50g. 7 times the amount of water (350ml water), 1% glucose aqueous solution (3.5g glucose was added to 350ml), 1% sucrose aqueous solution (3.5g sucrose was added to 350ml), 1% maltose aqueous solution (3.5g maltose was added to 350ml) was added respectively, soaked for 30min, refluxed for 30min, and then filtered with gauze while hot to obtain different sugar-Astragalus extract.

[0135] 2.2 Astragaloside, calycosin-7-glucoside, calycosin and formononetin content determination

[0136] 2.2.1 Instruments and reagents

[0137] Agilent 1260 high performance liquid chromatograph with DAD detector (Agilent, USA); Agilent 1200 high performance liquid chromatograph with ELSD detector (Agilent, USA); DZKW-4 electronic constant-temperature water bath (Beijing Zhongxingweiye Instrument Co., Ltd.); DK-98-IIA electric heating constant-temperature water bath (Taisite Instrument Co., Ltd. of Tianjin); ME204 / 02 electronic balance [Mettler-Toledo Instrument (Shanghai) Co., Ltd., one hundred thousandth balance]; KQ-250DE numerical control ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); HC-3018 high-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.); solid phase extraction column UniEIut C18 EC column (1000mg / 6ml) (Huapukexue Beijing Technology Co., Ltd.).

[0138] Reference substances Astragaloside IV (batch number CHB170727), Calycosin-7-glucoside (batch number CHB161105), Vitexin (batch number CHB150517), and Calycosin (batch number CHB161104) were purchased from Chengdu Keluoma Biological Technology Co., Ltd., with a purity of HPLC≥98%; acetonitrile (chromatographic pure, Thermo Fisher Scientific), methanol (chromatographic pure, Thermo Fisher Scientific), and formic acid (analytical pure) were purchased from National Pharmaceutical Group Chemical Reagent Co., Ltd. Water was Wahaha pure water.

[0139] 2.2.2 Preparation of test solution

[0140] Preparation of test solution of saponin components: 5 ml of different sugar-Astragalus extract was taken, 20 ml of 60% methanol was added, and ultrasonic mixing was performed. Then, 10 ml of ammonia water was added, mixed uniformly, centrifuged at 3600 r / min for 10 min, and the supernatant was passed through a Huapu UniEIut C18 EC column (1000mg / 6ml). Then, 10 ml of pure water was used for elution, and 5 ml of methanol was used for elution. The methanol eluate was collected, centrifuged at 8000 r / min for 5 min, and the supernatant was passed through a 0.45μm microporous filter membrane to obtain the test solution.

[0141] Preparation of test solution of flavonoid components: 300μl of different sugar-Astragalus extract was taken, 700μl of water was added, and the mixture was placed in a 1.5ml centrifuge tube. Then, the mixture was centrifuged at 12000 r / min for 5 min, and the supernatant was passed through a 0.45μm microporous filter membrane to obtain the test solution.

[0142] 2.2.3 Preparation of reference solution

[0143] Take astragaloside, calycosin-7-glucoside, ononin and calycosin reference substance, accurately weigh, add methanol to prepare 1 mg / ml stock solution. Dilute the stock solution into different series of concentrations to establish the working curve.

[0144] The linear equation of astragaloside is lnY = 1.3271lnX + 9.9314, r = 1.0000; the linear equation of calycosin-7-glucoside is Y = 35129X + 14, r = 0.9998; the linear equation of ononin is Y = 10024X + 33, r = 0.9998; the linear equation of calycosin is Y = 4656X + 25, r = 0.9995.

[0145] 2.2.4 Content determination method

[0146] The content determination method of saponin components is analyzed by Agilent 1200 type liquid chromatograph (with ELSD detector), the chromatographic column is Thermo Acclaim 120 C18 (250x4.6mm, 5um), the mobile phase is 0.1% formic acid water (A) to 0.1% formic acid acetonitrile (B), gradient elution: 0-5min, 5%-10%B; 5-10min, 10%-32%B; 10-30min, 32%-45%B; 30-35min, 45%-95%B; 35-40min, 95%-20%B), column temperature: room temperature, flow rate 1ml / min, drift tube temperature 100℃, carrier gas flow rate 2.5l / min, sample size 20ul.

[0147] The content determination method of flavonoid components is analyzed by Agilent 1260 type high performance liquid chromatograph (with DAD detector), the chromatographic column is Thermo Acclaim 120 C18 (250x4.6mm, 5um), the mobile phase is 0.1% formic acid aqueous solution (A) and 0.1% formic acid acetonitrile (B), gradient elution: 0-8min, 5%-20%B; 8-15min, 20%-25%B; 15-20min, 25%B; 20-30min, 25%-40%B; 30-40min, 40%-60%B; flow rate 1ml / min; detection wavelength 260nm; column temperature 25℃; sample size 10ul.

[0148] 2.3 Experimental results

[0149] The content determination results of effective components in different sugar- astragalus extract solutions are shown in Table 2.

[0150] Table 2 Content determination results of effective components in different sugar- astragalus extract solutions

[0151]

[0152] *P<0.05 VS water

[0153] As can be seen from Table 2, glucose, sucrose and maltose (1 g / 100 ml concentration) can all increase the content of effective components in Huangqi to different extents, and the effect of sucrose is the most obvious.

[0154] Example Three: Experiment of extracting raw Huangqi decoction pieces by different sugar solutions

[0155] 3.1 Preparation method of different sugar-Huangqi extraction solutions (extracted twice with 1% sugar)

[0156] Take 4 portions (HQ, HQ+1% sucrose, HQ+1% maltose, HQ+1% lactose) of the same batch of raw Huangqi decoction pieces (from Lingchuan, Shanxi) with each portion being 50 g. Add 7 times the amount of water (350 ml water), 1% sucrose aqueous solution (3.5 g sucrose added to water to make up to 350 ml), 1% maltose aqueous solution (3.5 g maltose added to water to make up to 350 ml), and 1% lactose aqueous solution (3.5 g lactose added to water to make up to 350 ml) to each portion respectively, soak for 30 min, reflux for 30 min, filter while hot with gauze, and then add 6 times the amount of water (300 ml water), 1% sucrose aqueous solution (3.0 g sucrose added to water to make up to 300 ml), 1% maltose aqueous solution (3.0 g maltose added to water to make up to 300 ml), and 1% lactose aqueous solution (3.0 g lactose added to water to make up to 300 ml) to the drug residues respectively, reflux for 20 min, filter while hot, and then combine the two filtrates to obtain different sugar-Huangqi extraction solutions.

[0157] 3.2 Determination of the contents of calycosin-7-glucoside, calycosin, formononetin and formononetin-7-glucoside

[0158] 3.2.1 Instruments and reagents

[0159] Agilent 1260 type high performance liquid chromatograph equipped with a DAD detector (Agilent, USA); Agilent 1200 type high performance liquid chromatograph equipped with an ELSD detector (Agilent, USA); DZKW-4 type electronic constant-temperature water bath (Beijing Zhongxingweiye Instrument Co., Ltd.); DK-98-IIA electric heating constant-temperature water bath (Tianjin Test Instrument Co., Ltd.); ME204 / 02 type electronic balance [Mettler-Toledo Instrument (Shanghai) Co., Ltd., 1 / 10,000 balance]; KQ-250DE type numerical control ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); HC-3018 high-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.); solid-phase extraction column UniEIut C18 EC column (1000 mg / 6 ml) (Huapukexue Beijing Technology Co., Ltd.).

[0160] The reference substance Astragaloside IV (batch number CHB170727), Calycosin-7-glucoside (batch number CHB161105), Maackiaamine (batch number CHB150517), and Calycosin (batch number CHB161104) were purchased from Chengdu Keluoma Biological Technology Co., Ltd., with a purity of HPLC≥98%; acetonitrile (chromatographic pure, Thermo Fisher Scientific), methanol (chromatographic pure, Thermo Fisher Scientific), and formic acid (analytical pure) were purchased from National Pharmaceutical Group Chemical Reagent Co., Ltd. Water was Wahaha pure water.

[0161] 3.2.2 Preparation of test solution

[0162] Preparation of saponin test solution: 5 ml of different sugar-Astragalus extract was taken, 20 ml of 60% methanol was added, and ultrasonic mixing was performed. Then, 10 ml of ammonia water was added, mixed well, and centrifuged at 3600 r / min for 10 min. The supernatant was passed through a Waters UniEIut C18 EC column (1000 mg / 6 ml), eluted with 10 ml of pure water, and then eluted with 5 ml of methanol. The methanol eluate was collected, centrifuged at 8000 r / min for 5 min, and the supernatant was passed through a 0.45 μm microporous filter membrane to obtain the test solution.

[0163] Preparation of flavonoid test solution: 300 μl of different sugar-Astragalus extract was taken, 700 μl of water was added, and centrifuged at 12000 r / min for 5 min. The supernatant was passed through a 0.45 μm microporous filter membrane to obtain the test solution.

[0164] 3.2.3 Preparation of reference solution

[0165] Astragaloside IV, Calycosin-7-glucoside, Maackiaamine, and Calycosin reference substances were accurately weighed and dissolved in methanol to prepare a 1 mg / ml stock solution. The stock solution was diluted to different concentrations to establish a working curve.

[0166] The linear equation of Astragaloside IV was lnY = 1.3271 lnX + 9.9314, r = 1.0000; the linear equation of Calycosin-7-glucoside was Y = 35129X + 14, r = 0.9998; the linear equation of Maackiaamine was Y = 10024X + 33, r = 0.9998; and the linear equation of Calycosin was Y = 4656X + 25, r = 0.9995.

[0167] 3.2.4 Content determination method

[0168] Saponins content determination method: Agilent 1200 type liquid chromatograph (with ELSD detector) was used for analysis, chromatographic column: Thermo Acclaim 120 C18 (250x4.6mm, 5μm); mobile phase: 0.1% formic acid water (A) ~ 0.1% formic acid acetonitrile (B), gradient elution: 0-5min, 5%-10%B; 5-10min, 10%-32%B; 10-30min, 32%-45%B; 30-35min, 45%-95%B; 35-40min, 95%-20%B), column temperature: room temperature, flow rate 1ml / min, drift tube temperature 100℃, carrier gas flow rate 2.5l / min, injection volume 20μl.

[0169] Flavonoids content determination method: Agilent 1260 type high performance liquid chromatograph (with DAD detector) was used for analysis, chromatographic column: Thermo Acclaim 120 C18 (250x4.6mm, 5μm); mobile phase: 0.1% formic acid aqueous solution (A) and 0.1% formic acid acetonitrile (B), gradient elution: 0-8min, 5%-20%B; 8-15min, 20%-25%B; 15-20min, 25%B; 20-30min, 25%-40%B; 30-40min, 40%-60%B; flow rate 1ml / min; detection wavelength 260nm; column temperature 25℃; injection volume 10μl.

[0170] 3.3 Experimental results

[0171] The content determination results of different sugar-Huangqi extractives are shown in Table 3.

[0172] Table 3 Content determination results of different sugar-Huangqi extractives

[0173]

[0174]

[0175] *P<0.05 VS water

[0176] As can be seen from Table 3, sucrose, maltose, lactose (1g / 100ml concentration) can all increase the content of active ingredients in Huangqi to different degrees, among which sucrose has the most obvious effect.

[0177] According to the results of Table 1 and Table 3, it can be seen that sucrose can significantly increase the extraction rate of main active ingredients in Huangqi, including flavonoids (calycosin-7-glucoside, ononin, calycosin) and saponins (astragaloside IV).

[0178] Example 4 Extraction of raw radix astragali pieces with different concentrations of sucrose solution

[0179] 4.1 Preparation of different concentrations of sucrose-radix astragali extraction solution (0-10% different concentrations of sucrose extraction)

[0180] The same batch of raw radix astragali pieces (from Lingchuan, Shanxi) was taken in six portions (HQ, HQ+0.25% sucrose, HQ+0.5% sucrose, HQ+1% sucrose, HQ+2% sucrose, HQ+10% sucrose), each of 50 g. Seven times the amount of water (350 ml water), 0.25% sucrose aqueous solution (0.9 g sucrose added to water to make up to 350 ml), 0.5% sucrose aqueous solution (1.8 g sucrose added to water to make up to 350 ml), 1% sucrose aqueous solution (3.5 g sucrose added to water to make up to 350 ml), 1.25% sucrose aqueous solution (4.4 g sucrose added to water to make up to 350 ml), 1.5% sucrose aqueous solution (5.3 g sucrose added to water to make up to 350 ml), 2% sucrose aqueous solution (7 g sucrose added to water to make up to 350 ml), and 10% sucrose aqueous solution (35 g sucrose added to water to make up to 350 ml) were added to each portion, respectively, soaked for 30 min, refluxed for 30 min, and filtered with gauze while hot. The residue was added with six times the amount of water (300 ml water), 0.25% sucrose aqueous solution (0.8 g sucrose added to water to make up to 300 ml), 0.5% sucrose aqueous solution (1.5 g sucrose added to water to make up to 300 ml), 1% sucrose aqueous solution (3.0 g sucrose added to water to make up to 300 ml), 1.25% sucrose aqueous solution (3.8 g sucrose added to water to make up to 300 ml), 1.5% sucrose aqueous solution (4.5 g sucrose added to water to make up to 300 ml), 2% sucrose aqueous solution (6.0 g sucrose added to water to make up to 300 ml), and 10% sucrose aqueous solution (30 g sucrose added to water to make up to 300 ml), respectively, refluxed for 20 min, filtered while hot, and the filtrates from the two times were combined to obtain different concentrations of sucrose-radix astragali extraction solution.

[0181] 4.2 Determination of the contents of astragaloside, calycosin-7-glucoside, maianhanside, and calycosin

[0182] 4.2.1 Instruments and reagents

[0183] Agilent 1260 high performance liquid chromatograph with DAD detector (Agilent, USA); Agilent 1200 high performance liquid chromatograph with ELSD detector (Agilent, USA); DZKW-4 electronic constant-temperature water bath (Beijing Zhongxingweiye Instrument Co., Ltd.); DK-98-IIA electric heating constant-temperature water bath (Tianjin Teset Instrument Co., Ltd.); ME204 / 02 electronic balance [Mettler-Toledo Instrument (Shanghai) Co., Ltd., one hundred-thousandth balance]; KQ-250DE digital ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.); HC-3018 high-speed centrifuge (Anhui Zhongke Zhongjia Scientific Instrument Co., Ltd.); solid phase extraction column UniEIut C18 EC column (1000 mg / 6 ml) (Huapukexue Beijing Technology Co., Ltd.).

[0184] The reference substances Astragaloside IV (batch number CHB170727), Calycosin-7-glucoside (batch number CHB161105), Ononin (batch number CHB150517), and Calycosin (batch number CHB161104) were purchased from Chengdu Keluoma Biological Technology Co., Ltd., with a purity of HPLC≥98%; acetonitrile (chromatographic pure, Thermo Fisher Scientific) and methanol (chromatographic pure, Thermo Fisher Scientific) were purchased from National Pharmaceutical Group Chemical Reagent Co., Ltd., and formic acid was analytical pure.

[0185] 4.2.2 Preparation of test solution

[0186] Preparation of test solution of saponin components: 5 ml of different sugar-Astragalus extract was taken, 20 ml of 60% methanol was added, and ultrasonic mixing was performed, 10 ml of ammonia water was added, and mixed, and placed in a centrifuge at 3600 r / min for 10 min, and the supernatant was passed through a Huapu UniEIut C18 EC column (1000 mg / 6 ml), eluted with 10 ml of pure water, and then eluted with 5 ml of methanol, and the methanol eluate was collected, centrifuged at 8000 r / min for 5 min, and the supernatant was passed through a 0.45 μm microporous filter membrane to obtain the test solution.

[0187] Preparation of test solution of flavonoid components: 300 μl of different sugar-Astragalus extract was taken, 700 μl of water was added, and placed in a 1.5 ml centrifuge tube, centrifuged at 12000 r / min for 5 min, and the supernatant was passed through a 0.45 μm microporous filter membrane to obtain the test solution.

[0188] 4.2.3 Preparation of reference solution

[0189] Astragaloside IV, Calycosin-7-glucoside, Ononin, and Calycosin reference substances were accurately weighed and dissolved in methanol to prepare a 1 mg / ml stock solution. The stock solution was diluted to different concentrations to establish a working curve.

[0190] The linear equation of astragaloside is lnY = 1.3271lnX + 9.9314, r = 1.0000; the linear equation of calycosin-7-glucoside is Y = 35129X + 14, r = 0.9998; the linear equation of ononin is Y = 10024X + 33, r = 0.9998; the linear equation of calycosin is Y = 4656X + 25, r = 0.9995.

[0191] 4.2.4 Content determination method

[0192] The content determination method of saponin components is as follows: Agilent 1200 type liquid chromatograph (with ELSD detector) is used for analysis, the chromatographic column is Thermo Acclaim 120 C18 (250*4.6mm, 5μm), the mobile phase is 0.1% formic acid water (A) to 0.1% formic acid acetonitrile (B), gradient elution: 0-5min, 5%-10%B; 5-10min, 10%-32%B; 10-30min, 32%-45%B; 30-35min, 45%-95%B; 35-40min, 95%-20%B, the column temperature is room temperature, the flow rate is 1ml / min, the drift tube temperature is 100℃, the carrier gas flow rate is 2.5l / min, and the sample injection amount is 20μl.

[0193] The content determination method of flavonoid components is as follows: Agilent 1260 type high performance liquid chromatograph (with DAD detector) is used for analysis, the chromatographic column is Thermo Acclaim 120 C18 (250*4.6mm, 5μm), the mobile phase is 0.1% formic acid water solution (A) and 0.1% formic acid acetonitrile (B), gradient elution: 0-8min, 5%-20%B; 8-15min, 20%-25%B; 15-20min, 25%B; 20-30min, 25%-40%B; 30-40min, 40%-60%B, the flow rate is 1ml / min, the detection wavelength is 260nm, the column temperature is 25℃, and the sample injection amount is 10μl.

[0194] 4.3 Experimental results

[0195] The content determination results of effective components in the astragalus extract obtained by different concentrations of sucrose solution extracting raw astragalus slices are shown in Table 4.

[0196] Table 4 Content determination results of effective components in the astragalus extract obtained by different concentrations of sucrose

[0197]

[0198] *P<0.05 VS water

[0199] As can be seen from Table 4, the sucrose concentration in the range of 0.25 g / 100 ml to 1.25 g / 100 ml can improve the content of effective components in Huangqi in different degrees, and the effect is most obvious when the sucrose concentration is in the range of 1 g / 100 ml to 1.25 g / 100 ml. In addition, the sucrose concentration of 10 g / 100 ml can also improve the content of effective components in Huangqi.

[0200] The above is only the embodiment of the present application, and the common knowledge of specific structure and characteristics in the scheme is not described in detail. It should be pointed out that for those skilled in the art, without departing from the present application, a number of modifications and improvements can also be made, which should also be considered as the protection scope of the present application, which will not affect the effect and practicality of the patent. The protection scope of the present application should be subject to the content of its claims, and the specific implementation mode in the specification can be used to explain the content of the claims.

[0201] The above has carried on the detailed introduction to the embodiment of the present application, and the principle and implementation mode of the present application are described by applying specific examples. The above embodiment is only used to help understand the method and core idea of the present application. At the same time, the changes or deformations made by those skilled in the art according to the idea of the present application, based on the specific implementation mode and application range of the present application, all belong to the protection scope of the present application. In summary, the content of the specification should not be understood as the limitation of the present application.

Claims

1. A method for increasing the extraction of effective components from Astragalus membranaceus by using sugar, characterized in that, The method comprises the following steps: (1) preparing a sugar solution with a concentration of 1 g / 100 mL; (2) soaking the Astragalus membranaceus in the sugar solution; and (3) heating the Astragalus membranaceus in the sugar solution to extract the Astragalus membranaceus once, and obtaining Astragalus membranaceus extract liquid through filtration; wherein the sugar is sucrose; wherein the effective components consist of Astragaloside IV, Calycosin-7-glucoside, Ononin and Calycosin.

2. A method for increasing the extraction of effective components of Astragalus membranaceus by using sugar, characterized in that, The method comprises the following steps: (1) preparing a sugar solution with a concentration of 1 g / 100 mL; (2) soaking the Astragalus membranaceus in the sugar solution; (3) heating the Astragalus membranaceus in the sugar solution to extract the Astragalus membranaceus, and obtaining first Astragalus membranaceus extract liquid through filtration; (4) adding the sugar solution to the Astragalus membranaceus, heating the Astragalus membranaceus to extract the Astragalus membranaceus, and obtaining second Astragalus membranaceus extract liquid; and (5) combining the first Astragalus membranaceus extract liquid and the second Astragalus membranaceus extract liquid to obtain Astragalus membranaceus extract liquid; wherein the sugar is sucrose, fructose or trehalose; wherein the effective components consist of Astragaloside IV, Calycosin-7-glucoside and Ononin.

3. The method according to claim 1 or 2, characterized in that, In step (2), the soaking is room temperature soaking.

4. The method of claim 3, wherein, The step of soaking comprises soaking the Astragalus membranaceus in the sugar solution at room temperature for 10-120 min.

5. The method of claim 4, wherein, The step of soaking comprises soaking the Astragalus membranaceus in the sugar solution at room temperature for 20-60 min.

6. The method of claim 5, wherein, The step of soaking comprises soaking the Astragalus membranaceus in the sugar solution at room temperature for 28.5 min to 31.5 min.

7. The method according to claim 1 or 2, characterized in that, In step (2), the weight-to-volume ratio of the Astragalus membranaceus to the sugar solution is 1:1-1:

50.

8. The method of claim 7, wherein, The weight-to-volume ratio of the Astragalus membranaceus to the sugar solution is 1:1-1:

20.

9. The method of claim 8, wherein, The weight-to-volume ratio of the Astragalus membranaceus to the sugar solution is 1:6-1:

7.

10. The method of claim 1 or 2, wherein, In step (3), the heating extraction is heating reflux extraction.

11. The method of claim 10, wherein, The heating time is 20 min to 60 min.

12. The method of claim 11, wherein, The heating time is 28.5 min to 31.5 min.

13. Use of the method according to any one of claims 1 to 12 in the preparation of Astragalus membranaceus extract liquid or Astragalus membranaceus concentrated liquid.

14. Use of the method according to any one of claims 1 to 12 in the preparation of a pharmaceutical composition containing Astragalus membranaceus.

15. Use of the method according to any one of claims 1 to 12 in the preparation of a pharmaceutical preparation or functional food or health food containing Astragalus membranaceus.

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