A method for simultaneous extraction of multi-components from Polygonatum sibiricum

Abstract

The present invention belongs to the field of chemical extraction, and in particular, relates to a method for simultaneous extraction of multiple components of polygonatum. The method comprises: 1) taking a polygonatum extract, performing alcohol precipitation separation to obtain polysaccharides, and obtaining a pre-extracted solution after extracting the polysaccharides; 2) the pre-extracted solution after separating the polysaccharides is adsorbed by a macroporous adsorption column, and eluted to obtain a flavonoid solution and a saponin solution, respectively, that is, to achieve the extraction of flavonoids and saponins. The present invention provides a method for efficiently separating polysaccharides, flavonoids and saponins in a polygonatum extract, which can significantly and efficiently achieve effective separation and purification of these three components to obtain a high-purity product, and the overall method is simple and efficient, and easy to implement industrially.

Description

Technical Field

[0001] The invention belongs to the field of chemical extraction, and in particular relates to a multi-component synchronous extraction method of polygonatum. Background Art

[0002] Polygonatum sibiricum is a perennial herbaceous plant of the Liliaceae family and the Polygonatum genus. It is an important Chinese medicinal material with both medicinal and edible properties. It contains a variety of active ingredients such as polysaccharides, saponins, flavonoids, and polyphenols. Studies have shown that Polygonatum sibiricum polysaccharides have the effects of lowering blood sugar, anti-aging, lowering blood lipids, and enhancing memory; Polygonatum sibiricum saponins have the effects of lowering blood sugar, anti-tumor, and anti-oxidation; the biological activities of Polygonatum sibiricum flavonoids mainly include anti-fatigue, vascular protection, lowering blood sugar, lowering blood lipids, anti-aging, and antibacterial effects. Therefore, the extraction, separation, and purification of active ingredients such as polysaccharides, steroidal saponins, and flavonoids in Polygonatum sibiricum have important economic significance and production value.

[0003] However, in actual production operations, the extraction process of active ingredients from polygonatum is inefficient or costly. Usually, the extraction method is used, which requires multiple positive and negative extractions to separate and extract multiple components such as polygonatum polysaccharides, flavonoids and saponins, while other processes are difficult to achieve effective separation of various components. Therefore, it is of great industrial value to develop a more concise and efficient method that is conducive to the simultaneous extraction and separation of multiple components of polygonatum. Summary of the invention

[0004] In order to solve the problems that the existing extraction process is relatively complicated or it is difficult to extract and separate different components in polygonatum, the present invention provides a method for simultaneous extraction of multiple components of polygonatum.

[0005] The objects of the present invention are:

[0006] 1. Improve the effective extraction of the three major components of Polygonatum sibiricum;

[0007] 2. Improve extraction efficiency and ensure extraction effect;

[0008] 3. Ensure the high yield of each component.

[0009] To achieve the above objectives, the present invention adopts the following technical solutions.

[0010] A method for synchronously extracting multiple components of polygonatum,

[0011] The method comprises:

[0012] 1) taking a polygonatum extract, performing alcohol precipitation separation to obtain polysaccharides, and obtaining a pre-extracted solution after the polysaccharides are extracted;

[0013] 2) The pre-extracted liquid after the polysaccharide is separated is adsorbed by a macroporous adsorption column, and after elution, a flavonoid liquid and a saponin liquid are obtained respectively, that is, the flavonoids and saponins are extracted separately.

[0014] Preferably, in step 1), the polygonatum extract is prepared by mixing defatted polygonatum powder with water in a ratio of 1:15-25, and then extracting and concentrating the mixture in a water bath and collecting the upper aqueous phase, i.e., the polygonatum extract.

[0015] Preferably, in step 2), the pre-extracted liquid is adsorbed on a macroporous adsorption column until the macroporous adsorption column is saturated with adsorption, and then flavonoids and saponins are eluted in sequence.

[0016] As a preference,

[0017] The flavonoids are eluted using a low alcohol eluent;

[0018] The low alcohol eluent is an alcohol aqueous solution and / or water;

[0019] The alcohol content in the alcohol aqueous solution is ≤30% VOL.

[0020] As a preference,

[0021] The flavonoid elution end point is that the flavonoid content in the eluate is ≤ 2 wt %.

[0022] As a preference,

[0023] The saponin is eluted using a high concentration alcohol solution;

[0024] The high concentration alcohol solution is an aqueous solution of alcohol, wherein the alcohol content is ≥50wt%.

[0025] As a preference,

[0026] The elution end point is that the saponin content in the eluate is ≤ 2 wt %.

[0027] Preferably, the pre-extracted liquid is first mixed with water to obtain a diluent, and then the diluent is quantitatively passed through a macroporous adsorption column for adsorption, and the effluent of the diluent is recovered to obtain a flavonoid solution, which is then eluted with an organic alcohol aqueous solution with an alcohol content of ≥50wt%, and the eluent is recovered to obtain a saponin solution.

[0028] As a preference,

[0029] The pre-extracted liquid is mixed with water having a volume of at least 19 times that of the pre-extracted liquid to obtain a diluent;

[0030] When the diluent is adsorbed by the macroporous adsorption column, the ratio of the volume of the diluent used in a single time to the filling volume of the adsorption filler in the macroporous adsorption column is controlled to be (2-5):1.

[0031] As a preference,

[0032] The filler in the macroporous adsorption column is AB-8 macroporous adsorption resin.

[0033] The technical solution of the present invention mainly provides two multi-component synchronous extraction and separation methods.

[0034] One of them is the alcohol precipitation-double-layer chromatography two-step method. In this method, the pre-treated polygonatum powder is first used as the raw material to prepare the polygonatum extract, in which the main components are polysaccharides, flavonoids and saponins. This method has relatively low requirements for polygonatum powder. It can simply use the commercially available, un-impurified coarse polygonatum residue powder as the raw material to prepare the extract. The effective ingredients in the polygonatum powder are dissolved in the solution system. Since polygonatum polysaccharides are insoluble in alcohol, high-purity polysaccharides can be separated very efficiently and thoroughly. Then, through the cooperation of a macroporous adsorption column, the target component is adsorbed on the macroporous adsorption column, and then different eluents are used to elute the target components separately. During the experiment, a saturated single-column single elution test was also carried out, and it was found that Figure 1 As shown, the elution transfer of flavonoids at low alcohol concentration is targeted, and the elution of flavonoids can be achieved; and after the elution of flavonoids, the saponins can be further eluted in a targeted manner, achieving effective separation of the two and removal of impurities. Compared with multiple forward and reverse extractions, the efficiency is significantly improved.

[0035] The second method is an alcohol precipitation-single chromatography high-efficiency two-step method. In this method, the residue of Polygonatum odoratum can also be used as a raw material for preparation, but it needs to be deproteinized and defatted to remove impurities; then the effective ingredients in the Polygonatum odoratum powder are dissolved in the solution system. Since Polygonatum odoratum polysaccharide is insoluble in alcohol, it can be very efficient and thoroughly separated from high-purity polysaccharides. Then, through the cooperation of a macroporous adsorption column, flavonoids and saponins can be directly separated when passing through a macroporous adsorption resin. Because the invention researchers of this application found in the previous experimental process that in the actual adsorption process, by controlling the substance concentration of the adsorbed liquid, completely different adsorption effects can be produced, thereby achieving a single adsorption chromatography separation of the two. In this process, the eluent is quickly passed through a macroporous adsorption column (chromatographic column) to keep it in an unsaturated adsorption state, while the saponins in the eluent are adsorbed on the chromatographic column, and the flavonoids flow downstream and are almost not adsorbed, achieving direct separation, which can very efficiently improve the efficiency of industrial implementation. Compared with the first method, it is more thorough and effective in separating flavonoids and saponins.

[0036] As for the above two methods, the type of macroporous adsorption resin in the chromatography column selected by the technical solution of the present invention will have a greater impact on the results. For example, common macroporous adsorption resin brands include AB-8, D101, D201 and D301, which can generally be replaced equally. However, for the technical solution of the present invention, a slight change in the adsorption performance of the macroporous adsorption resin may be significantly amplified, resulting in a significant difference.

[0037] The beneficial effects of the present invention are:

[0038] The present invention provides a method for efficiently separating polysaccharides, flavonoids and saponins in a polygonatum extract, which can significantly and efficiently achieve effective separation and purification of the three components to obtain a high-purity product, and the overall method is simple, efficient and easy to implement industrially. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 This is a graph showing the test results of the saturated single column-single elution transfer rate of flavonoids and saponins based on the alcohol-water concentration ratio. DETAILED DESCRIPTION

[0040] The present invention is further described in detail below in conjunction with specific embodiments and the accompanying drawings. A person of ordinary skill in the art will be able to implement the present invention based on these descriptions. In addition, the embodiments of the present invention involved in the following description are generally only embodiments of a part of the present invention, rather than all embodiments. Therefore, based on the embodiments of the present invention, all other embodiments obtained by a person of ordinary skill in the art without making creative work should fall within the scope of protection of the present invention.

[0041] Unless otherwise specified, the raw materials used in the examples of the present invention are all commercially available or available to those skilled in the art; unless otherwise specified, the methods used in the examples of the present invention are all methods known to those skilled in the art.

[0042] Unless otherwise specified, the coarse powder of polygonatum used in the embodiment of the present invention is prepared by the following process: the waste residue of polygonatum is dried at 45°C to constant weight, then crushed by a grinder, passed through a 60-mesh sieve, the crushed sample is weighed, anhydrous ethanol is added according to a solid-liquid ratio of 1g:8mL, soaked for 48h, filtered, and dried to obtain a defatted sample, i.e., coarse powder of polygonatum, i.e., defatted polygonatum powder.

[0043] Unless otherwise specified, the polygonatum powder used in the embodiments of the present invention is prepared by the following process: take the coarse polygonatum powder and add deionized water at a solid-liquid ratio of 1g:20mL, water bath at 75°C for 2.5h, centrifuge to obtain the supernatant, concentrate to 35% of the original volume, remove protein by Sevage method, and then freeze-dry to obtain the polygonatum powder.

[0044] Example 1

[0045] A method for synchronously extracting multi-components from polygonatum, the method specifically comprising:

[0046] 1) Mix the coarse powder of Polygonatum sibiricum with deionized water at a solid-liquid ratio of 1g:20mL, and perform constant temperature extraction in a water bath at 75℃

[0047] 2.5 h, then concentrated to 30% of the original volume, and the aqueous phase solution was collected as the polygonatum sibiricum extract;

[0048] 2) Mix the polygonatum extract with anhydrous ethanol at a volume ratio of 1:4, precipitate overnight at 4°C, recover the precipitated polysaccharide sample by centrifugation, and take the supernatant to obtain a pre-extracted solution;

[0049] 3) The pre-extracted liquid is subjected to adsorption treatment by a chromatography column, wherein the chromatography column used is an AB-8 macroporous adsorption resin column, and after the chromatography column is adsorbed to saturation, or when flavonoids or saponins cannot be detected in the adsorption liquid, elution is performed respectively:

[0050] 3-1) Firstly, the chromatography column is eluted with 10% vol. ethanol aqueous solution, and the eluate is rapidly characterized. When the flavonoid content in the adsorbed liquid is ≤2 wt%, the elution end point is reached, and the eluate is combined, concentrated, and freeze-dried to obtain a flavonoid sample;

[0051] 3-2) The chromatography column was then eluted with 50% vol. ethanol aqueous solution, and the eluate was rapidly characterized until the saponin content in the eluate was ≤ 2 wt % to reach the elution end point. The eluate was combined, concentrated, and freeze-dried to obtain a saponin sample.

[0052] The polysaccharide recovery rate, flavonoid recovery rate, saponin recovery rate, sample (ele) purity, flavonoid transfer rate and saponin transfer rate of this example were characterized and calculated respectively, wherein the sample (ele) purity was characterized by XRD.

[0053] in:

[0054] The recovery rate is calculated as follows:

[0055]

[0056] Where: Rec ele Represents the recovery rate of the corresponding sample (ele), dimensionless, M ele Indicates the total recovery (mass) of the corresponding sample (ele), in g, c ele Indicates the purity of the corresponding sample (ele), dimensionless, Q ele It represents the content (mass) of the corresponding sample (ele) in the coarse powder of Polygonatum sibiricum, in g. The subscript ele represents the sample, which is a polysaccharide, flavonoid or saponin.

[0057] The transfer rate is calculated as follows:

[0058]

[0059] T ele Represents the transfer rate of the corresponding sample (ele), dimensionless, M ele ′ represents the single column recovery (mass) of the corresponding sample (ele), which is the total amount of multiple recovery in a single column, in g, c ele Indicates the purity of the corresponding sample (ele), dimensionless, Q ele ' represents the content (mass) of the corresponding sample (ele) in the saturated chromatography column before elution, in g, and the subscript ele represents the sample, which is a polysaccharide, flavonoid or saponin.

[0060] After calculation, the recovery rate, purity and transfer rate of each sample in this example are shown in the following table.

[0061] Polysaccharides Flavonoids Saponin Recovery rate 98.6％ 91.2％ 93.2％ purity 99.3％ 98.1％ 99.6％ Transfer rate / 98.6％ 99.3％

[0062] It can be seen that the present invention can efficiently separate and extract polysaccharides, flavonoids and saponins in polygonatum, the recovery rate can reach more than 90%, and the purity can basically reach more than 98%, and the main impurity component in flavonoids is saponin.

[0063] Example 2

[0064] A method for synchronously extracting multi-components from polygonatum, the method specifically comprising:

[0065] 1) Mixing the coarse powder of polygonatum sibiricum with deionized water at a solid-liquid ratio of 1 g: 20 mL, performing constant temperature extraction in a water bath at 75° C. for 2.5 h, and then concentrating to 30% of the original volume, collecting the aqueous phase solution as the polygonatum sibiricum extract;

[0066] 2) Mix the polygonatum extract with anhydrous ethanol at a volume ratio of 1:4, precipitate overnight at 4°C, recover the precipitated polysaccharide sample by centrifugation, and take the supernatant to obtain a pre-extracted solution;

[0067] 3) The pre-extracted liquid is subjected to adsorption treatment by a chromatography column, wherein the chromatography column used is an AB-8 macroporous adsorption resin column, and after the chromatography column is adsorbed to saturation, or when flavonoids or saponins cannot be detected in the adsorption liquid, elution is performed respectively:

[0068] 3-1) Firstly, the chromatography column is eluted with 30% vol. ethanol aqueous solution, and the eluate is rapidly characterized. When the flavonoid content in the adsorbed liquid is ≤2 wt%, the elution end point is reached, and the eluate is combined, concentrated, and freeze-dried to obtain a flavonoid sample;

[0069] 3-2) The chromatography column was then eluted with 80% vol. ethanol aqueous solution, and the eluate was rapidly characterized until the saponin content in the eluate was ≤ 2 wt % to reach the elution end point. The eluate was combined, concentrated, and freeze-dried to obtain a saponin sample.

[0070] The polysaccharide recovery rate, flavonoid recovery rate, saponin recovery rate, sample (ele) purity, flavonoid transfer rate and saponin transfer rate of this example were characterized and calculated respectively, wherein the sample (ele) purity was characterized by XRD.

[0071] in:

[0072] The recovery rate is calculated as follows:

[0073]

[0074] Where: Rec ele Represents the recovery rate of the corresponding sample (ele), dimensionless, M ele Indicates the total recovery (mass) of the corresponding sample (ele), in g, c ele Indicates the purity of the corresponding sample (ele), dimensionless, Q ele It represents the content (mass) of the corresponding sample (ele) in the coarse powder of Polygonatum sibiricum, in g. The subscript ele represents the sample, which is a polysaccharide, flavonoid or saponin.

[0075] The transfer rate is calculated as follows:

[0076]

[0077] T ele Represents the transfer rate of the corresponding sample (ele), dimensionless, M ele ′ represents the single column recovery (mass) of the corresponding sample (ele), which is the total amount of multiple recovery in a single column, in g, c ele Indicates the purity of the corresponding sample (ele), dimensionless, Q ele ' represents the content (mass) of the corresponding sample (ele) in the saturated chromatography column before elution, in g, and the subscript ele represents the sample, which is a polysaccharide, flavonoid or saponin.

[0078] After calculation, the recovery rate, purity and transfer rate of each sample in this example are shown in the following table.

[0079] Polysaccharides Flavonoids Saponin Recovery rate 98.8％ 92.5％ 92.9％ purity 99.5％ 97.3％ 99.8％ Transfer rate / 99.7％ 98.1％

[0080] It can be seen that the present invention can efficiently separate and extract polysaccharides, flavonoids and saponins in Polygonatum sibiricum, and the recovery rate can reach more than 90%. Compared with Example 1, increasing the alcohol concentration used for the two elutions can improve the actual elution transfer rate of flavonoids and saponins to a certain extent. However, in the calculation process of the above table, since the saponin part of the flavonoids cannot be included in the calculation, the calculated transfer rate decreases, but the overall transfer effect is still relatively outstanding.

[0081] Example 3

[0082] A method for synchronously extracting multi-components from polygonatum, the method specifically comprising:

[0083] 1) Mixing the coarse powder of polygonatum sibiricum with deionized water at a solid-liquid ratio of 1 g: 20 mL, performing constant temperature extraction in a water bath at 75° C. for 2.5 h, and then concentrating to 30% of the original volume, collecting the aqueous phase solution as the polygonatum sibiricum extract;

[0084] 2) Mix the polygonatum extract with anhydrous ethanol at a volume ratio of 1:4, precipitate overnight at 4°C, recover the precipitated polysaccharide sample by centrifugation, and take the supernatant to obtain a pre-extracted solution;

[0085] 3) The pre-extracted liquid is subjected to adsorption treatment by a chromatography column, wherein the chromatography column used is an AB-8 macroporous adsorption resin column, and after the chromatography column is adsorbed to saturation, or when flavonoids or saponins cannot be detected in the adsorption liquid, elution is performed respectively:

[0086] 3-1) Firstly, the chromatography column is eluted with 30% vol. ethanol aqueous solution, and the eluate is rapidly characterized. When the flavonoid content in the adsorbed liquid is ≤2 wt%, the elution end point is reached, and the eluate is combined, concentrated, and freeze-dried to obtain a flavonoid sample;

[0087] 3-2) The chromatography column was then eluted with 95% vol. ethanol aqueous solution, and the eluate was rapidly characterized until the saponin content in the eluate was ≤ 2 wt % to reach the elution end point. The eluate was combined, concentrated, and freeze-dried to obtain a saponin sample.

[0088] The polysaccharide recovery rate, flavonoid recovery rate, saponin recovery rate, sample (ele) purity, flavonoid transfer rate and saponin transfer rate of this example were characterized and calculated respectively, wherein the sample (ele) purity was characterized by XRD.

[0089] in:

[0090] The recovery rate is calculated as follows:

[0091]

[0092] Where: Rec ele Represents the recovery rate of the corresponding sample (ele), dimensionless, M eleIndicates the total recovery (mass) of the corresponding sample (ele), in g, c ele Indicates the purity of the corresponding sample (ele), dimensionless, Q ele It represents the content (mass) of the corresponding sample (ele) in the coarse powder of Polygonatum sibiricum, in g. The subscript ele represents the sample, which is a polysaccharide, flavonoid or saponin.

[0093] The transfer rate is calculated as follows:

[0094]

[0095] T ele Represents the transfer rate of the corresponding sample (ele), dimensionless, M ele ′ represents the single column recovery (mass) of the corresponding sample (ele), which is the total amount of multiple recovery in a single column, in g, c ele Indicates the purity of the corresponding sample (ele), dimensionless, Q ele ' represents the content (mass) of the corresponding sample (ele) in the saturated chromatography column before elution, in g, and the subscript ele represents the sample, which is a polysaccharide, flavonoid or saponin.

[0096] After calculation, the recovery rate, purity and transfer rate of each sample in this example are shown in the following table.

[0097] Polysaccharides Flavonoids Saponin Recovery rate 98.3％ 92.6％ 93.1％ purity 99.7％ 97.1％ 99.1％ Transfer rate / 99.5％ 99.0％

[0098] It can be seen that the present invention can efficiently separate and extract polysaccharides, flavonoids and saponins in Polygonatum sibiricum, and the recovery rate can reach more than 90%. In the process of eluting saponins, a higher concentration of ethanol aqueous solution is used. It can be seen that the recovery and transfer effects are very excellent, but the purity of the product decreases to a certain extent, indicating that some residual flavonoids will be eluted with the high concentration of ethanol aqueous solution, resulting in a certain adverse effect on the purity of the product.

[0099] Comparative Example 1

[0100] A method for synchronously extracting multi-components from polygonatum, the method specifically comprising:

[0101] 1) Mixing the coarse powder of polygonatum sibiricum with deionized water at a solid-liquid ratio of 1 g: 20 mL, performing constant temperature extraction in a water bath at 75° C. for 2.5 h, and then concentrating to 30% of the original volume, collecting the aqueous phase solution as the polygonatum sibiricum extract;

[0102] 2) Mix the polygonatum extract with anhydrous ethanol at a volume ratio of 1:4, precipitate overnight at 4°C, recover the precipitated polysaccharide sample by centrifugation, and take the supernatant to obtain a pre-extracted solution;

[0103] 3) The pre-extracted liquid is subjected to adsorption treatment by a chromatography column, the chromatography column used is a D101 macroporous adsorption resin column, and after the chromatography column is adsorbed to saturation, or when flavonoids or saponins cannot be detected in the adsorption liquid, elution is performed respectively:

[0104] 3-1) Firstly, the chromatography column is eluted with 10% vol. ethanol aqueous solution, and the eluate is rapidly characterized. When the flavonoid content in the adsorbed liquid is ≤2 wt%, the elution end point is reached, and the eluate is combined, concentrated, and freeze-dried to obtain a flavonoid sample;

[0105] 3-2) The chromatography column was then eluted with 50% vol. ethanol aqueous solution, and the eluate was rapidly characterized until the saponin content in the eluate was ≤ 2 wt % to reach the elution end point. The eluate was combined, concentrated, and freeze-dried to obtain a saponin sample.

[0106] The polysaccharide recovery rate, flavonoid recovery rate, saponin recovery rate, sample (ele) purity, flavonoid transfer rate and saponin transfer rate of this example were characterized and calculated respectively, wherein the sample (ele) purity was characterized by XRD.

[0107] in:

[0108] The recovery rate is calculated as follows:

[0109]

[0110] Where: Rec ele Represents the recovery rate of the corresponding sample (ele), dimensionless, M ele Indicates the total recovery (mass) of the corresponding sample (ele), in g, c ele Indicates the purity of the corresponding sample (ele), dimensionless, Q ele It represents the content (mass) of the corresponding sample (ele) in the coarse powder of Polygonatum sibiricum, in g. The subscript ele represents the sample, which is a polysaccharide, flavonoid or saponin.

[0111] The transfer rate is calculated as follows:

[0112]

[0113] T ele Represents the transfer rate of the corresponding sample (ele), dimensionless, M ele ′ represents the single column recovery (mass) of the corresponding sample (ele), which is the total amount of multiple recovery in a single column, in g, c ele Indicates the purity of the corresponding sample (ele), dimensionless, Q ele ' represents the content (mass) of the corresponding sample (ele) in the saturated chromatography column before elution, in g, and the subscript ele represents the sample, which is a polysaccharide, flavonoid or saponin.

[0114] After calculation, the recovery rate, purity and transfer rate of each sample in this example are shown in the following table.

[0115] Polysaccharides Flavonoids Saponin Recovery rate 98.6％ 86.3％ 95.2％ purity 99.3％ 96.3％ 99.9％ Transfer rate / 95.2％ 96.9％

[0116] It can be seen that the present invention can efficiently separate and extract polysaccharides, flavonoids and saponins in polygonatum, the recovery rate can reach more than 90%, and the purity can basically reach more than 98%, and the main impurity component in flavonoids is saponin.

[0117] Compared with Example 1, this example only adjusted the type of adsorption column used, replacing the AB-8 macroporous adsorption column with the D101 adsorption column, but the results showed that the recovery rate and purity of flavonoids decreased significantly, while the recovery rate and purity of saponins increased. This was mainly due to the different binding strengths of the two components of different adsorption columns or the influence of factors such as the solution system, which made it easy for saponins to be taken down when flavonoids were eluted. It can be seen that for the technical solution of the present invention, the type of macroporous adsorption resin should not be adjusted at will. At least in the same test, the chromatography column of AB-8 macroporous adsorption resin has significantly different advantages compared with D101, D201 and D301, and is not as small as the influence of the chromatography column in the conventional chromatography method.

[0118] Example 4

[0119] A method for synchronously extracting multi-components from polygonatum, the method specifically comprising:

[0120] 1) Mixing the coarse powder of polygonatum sibiricum with deionized water at a solid-liquid ratio of 1 g: 20 mL, performing constant temperature extraction in a water bath at 75° C. for 2.5 h, and then concentrating to 30% of the original volume, removing protein impurities by the Sevage method, and collecting the aqueous phase solution as the polygonatum sibiricum extract;

[0121] 2) Mix the polygonatum extract with anhydrous ethanol at a volume ratio of 1:4, precipitate overnight at 4°C, recover the precipitated polysaccharide sample by centrifugation, and take the supernatant to obtain a pre-extracted solution;

[0122] 3) The pre-extracted liquid is mixed and diluted with deionized water in a volume ratio of 1:19 to obtain a diluent, and the diluent is adsorbed by a chromatography column, and the ratio of the volume of the diluent used for a single adsorption to the filling volume of the adsorption filler in the chromatography column is controlled to be 5:1. The chromatography column used is an AB-8 macroporous adsorption resin column, and the diluent effluent is directly recovered after the diluent passes through the chromatography column, and the diluent effluent is combined, concentrated and freeze-dried to obtain a flavonoid sample, and then the chromatography column is eluted with a 50% VOL ethanol aqueous solution, and the ratio of the eluent used in a single elution process to the filling volume of the adsorption filler in the chromatography column is 5:1, and the eluents are combined, concentrated and freeze-dried to obtain a saponin sample.

[0123] The polysaccharide recovery rate, flavonoid recovery rate, saponin recovery rate, sample (ele) purity, flavonoid transfer rate and saponin transfer rate of this example were characterized and calculated respectively, wherein the sample (ele) purity was characterized by XRD.

[0124] in:

[0125] The recovery rate is calculated as follows:

[0126]

[0127] Where: Rec ele Represents the recovery rate of the corresponding sample (ele), dimensionless, M ele Indicates the total recovery (mass) of the corresponding sample (ele), in g, c ele Indicates the purity of the corresponding sample (ele), dimensionless, Q ele It represents the content (mass) of the corresponding sample (ele) in the coarse powder of Polygonatum sibiricum, in g. The subscript ele represents the sample, which is a polysaccharide, flavonoid or saponin.

[0128] The transfer rate is calculated as follows:

[0129]

[0130] T ele Represents the transfer rate of the corresponding sample (ele), dimensionless, M ele ′ represents the single column recovery (mass) of the corresponding sample (ele), which is the total amount of multiple recovery in a single column, in g, c ele Indicates the purity of the corresponding sample (ele), dimensionless, Q ele ' represents the content (mass) of the corresponding sample (ele) in the saturated chromatography column before elution, in g, and the subscript ele represents the sample, which is a polysaccharide, flavonoid or saponin.

[0131] After calculation, the recovery rate, purity and transfer rate of each sample in this example are shown in the following table.

[0132] Polysaccharides Flavonoids Saponin Recovery rate 98.5％ 98.7％ 93.6％ purity 99.7％ 99.6％ 99.7％ Transfer rate / / 99.5％

[0133] Compared with Example 1, it can be clearly seen that the present invention changes the chromatography method and adopts direct current to directly recover flavonoids, which significantly improves the recovery rate and purity of flavonoids. This is because the loss of part of the residual liquid during the chromatography column chromatography process is reduced, and the purity of the three types of products can directly reach more than 99.5%, achieving the acquisition of high-purity products, and the overall process is more efficient. In addition, the recovery rate and purity of saponins are very effectively retained.

[0134] Example 5

[0135] A method for synchronously extracting multi-components from polygonatum, the method specifically comprising:

[0136] 1) Mixing the coarse powder of polygonatum sibiricum with deionized water at a solid-liquid ratio of 1 g: 20 mL, performing constant temperature extraction in a water bath at 75° C. for 2.5 h, and then concentrating to 30% of the original volume, removing protein impurities by the Sevage method, and collecting the aqueous phase solution as the polygonatum sibiricum extract;

[0137] 2) Mix the polygonatum extract with anhydrous ethanol at a volume ratio of 1:4, precipitate overnight at 4°C, recover the precipitated polysaccharide sample by centrifugation, and take the supernatant to obtain a pre-extracted solution;

[0138] 3) The pre-extracted liquid is mixed and diluted with deionized water in a volume ratio of 1:19 to obtain a diluent, and the diluent is adsorbed by a chromatography column, and the ratio of the volume of the diluent used for a single adsorption to the filling volume of the adsorption filler in the chromatography column is controlled to be 5:1. The chromatography column used is an AB-8 macroporous adsorption resin column, and the diluent effluent is directly recovered after the diluent passes through the chromatography column, and the diluent effluent is combined, concentrated and freeze-dried to obtain a flavonoid sample, and then the chromatography column is eluted with a 90% VOL ethanol aqueous solution, and the ratio of the eluent used in a single elution process to the filling volume of the adsorption filler in the chromatography column is 5:1, and the eluents are combined, concentrated and freeze-dried to obtain a saponin sample.

[0139] The polysaccharide recovery rate, flavonoid recovery rate, saponin recovery rate, sample (ele) purity, flavonoid transfer rate and saponin transfer rate of this example were characterized and calculated respectively, wherein the sample (ele) purity was characterized by XRD.

[0140] in:

[0141] The recovery rate is calculated as follows:

[0142]

[0143] Where: Rec ele Represents the recovery rate of the corresponding sample (ele), dimensionless, M ele Indicates the total recovery (mass) of the corresponding sample (ele), in g, c ele Indicates the purity of the corresponding sample (ele), dimensionless, Q ele It represents the content (mass) of the corresponding sample (ele) in the coarse powder of Polygonatum sibiricum, in g. The subscript ele represents the sample, which is a polysaccharide, flavonoid or saponin.

[0144] The transfer rate is calculated as follows:

[0145]

[0146] TR ele Represents the transfer rate of the corresponding sample (ele), dimensionless, Mele ′ represents the single column recovery (mass) of the corresponding sample (ele), which is the total amount of multiple recovery in a single column, in g, c ele Indicates the purity of the corresponding sample (ele), dimensionless, Q ele ' represents the content (mass) of the corresponding sample (ele) in the saturated chromatography column before elution, in g, and the subscript ele represents the sample, which is a polysaccharide, flavonoid or saponin.

[0147] After calculation, the recovery rate, purity and transfer rate of each sample in this example are shown in the following table.

[0148] Polysaccharides Flavonoids Saponin Recovery rate 98.2％ 99.0％ 95.0％ purity 99.8％ 99.5％ 99.8％ Transfer rate / / 99.8％

[0149] Compared with Example 4 and Example 3 and Example 1, it can be clearly seen that after changing the chromatography method and weakening the effect of flavonoid adsorption and retention on the high-concentration elution of saponins, using a higher concentration of ethanol to elute the saponins will no longer cause a decrease in purity, but will greatly improve the transfer rate and recovery rate, and can achieve better and more effective elution and recovery effects.

[0150] In this regard, a horizontal test study was conducted on Example 4 and Example 5, in which the main focus was on the dilution multiple of the pre-extraction solution and the ratio of the volume of the diluent used in a single time to the volume of the adsorption filler in the macroporous adsorption resin column.

[0151] In terms of the dilution multiple of the pre-extract, low dilution multiples cannot prevent the adsorption of flavonoids on the chromatography column. In Examples 4 and 5 of the present invention, the main core point is to make the chromatography column produce "selective" adsorption, and this selectivity is produced by dilution, and the chromatography column itself does not have selectivity. However, in the preliminary test, the following results were obtained: Figure 1 After the single elution test results shown, the inventors found that low-concentration ethanol aqueous solution and even pure water can elute flavonoids to a certain extent, while saponins are almost impossible to elute, thus actually creating a prospect for the development of "selective elution".

[0152] During the elution process, the following experiment was designed, and the saturated mounted chromatography column was eluted with several groups of flavonol aqueous solutions of different concentrations (the flavonoid content and the alcohol content were adjusted by two factors). The test results showed that when the flavonoid content and the ethanol content were both low, the adsorption and binding of the flavonoids to the chromatography column could be suppressed to a great extent, and the flavonoids could be effectively eluted by the chromatography column, thereby achieving a huge change in the chromatography method. The adsorbed liquid was directly used as the eluent, and the flavonoids were eluted while the saponin adsorption and mounting were performed, which could greatly improve the separation and extraction efficiency, and improve the product yield and purity. It has great use prospects and economic value for industrial production, and only needs to be pre-treated more fully to reduce the interference of impurities such as proteins.

Claims

1. A method for synchronous extraction of polygonatum multi-components, characterized in that: The extraction method comprises: 1) taking the polygonatum sibiricum extract after removing protein impurities, performing alcohol precipitation separation to obtain polysaccharides, and obtaining a pre-extracted solution after extracting the polysaccharides; 2) The pre-extracted liquid after the polysaccharide is separated is adsorbed by a macroporous adsorption column filled with AB-8, and after elution, flavonoid liquid and saponin liquid are obtained respectively, that is, the flavonoids and saponins are extracted separately; The elution method is: The pre-extracted liquid is first mixed with water to obtain a diluent, and then the diluent is quantitatively passed through a macroporous adsorption column for adsorption, and the diluent outflow is recovered to obtain a flavonoid solution, and then eluted with an organic alcohol aqueous solution with an alcohol content of ≥50 wt%, and the eluent is recovered to obtain a saponin solution; The pre-extracted liquid is mixed with water having a volume of at least 19 times that of the pre-extracted liquid to obtain a diluent; When the diluent is adsorbed by the macroporous adsorption column, the ratio of the volume of the diluent used in a single time to the filling volume of the adsorption filler in the macroporous adsorption column is controlled to be (2-5):

1.

2. The method for synchronously extracting multi-components from polygonatum according to claim 1, characterized in that: Step 1) The preparation method of the polygonatum extract is as follows: take defatted polygonatum powder and mix it with water in a ratio of 1g:15ml-25ml, perform water bath extraction and concentration in sequence, and then collect the upper aqueous phase liquid, i.e., the polygonatum extract.

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