A refined ginseng extract and its preparation method

Abstract

The present invention provides a method for preparing a refined ginseng extract, belonging to the technical field of removal of residual pesticides in plants. In the present invention, water, a flocculant and a crude ginseng extract are mixed for flocculation treatment, and after solid-liquid separation, the liquid material is collected; the liquid material is loaded onto a macroporous adsorption resin column, and then eluted successively with an alkali solution, water, an acid solution, water and a first ethanol aqueous solution, and the first ethanol aqueous solution eluate is collected; the first ethanol aqueous solution eluate is diluted with water, the obtained dilution is loaded onto a pesticide adsorption resin column, the column effluent is collected, and then eluted with a second ethanol aqueous solution, and the second ethanol aqueous solution eluate is collected; the column effluent and the second ethanol aqueous solution eluate are mixed, and concentrated and dried successively to obtain a refined ginseng extract. The method provided by the present invention can effectively improve the removal rate of residual pesticides in the ginseng extract, has simple operation and is suitable for large-scale industrial production.

Description

Technical Field

[0001] The present invention belongs to the technical field of removing residual pesticides from plants, and particularly relates to a refined ginseng extract and a preparation method thereof. Background Art

[0002] Ginseng is a plant of the Araliaceae family. Its roots, leaves, flowers, and fruits can all be eaten and used medicinally, and they have excellent efficacy and a large usage amount. Ginseng has changed from wild to cultivated, and its output has been increasing year by year. In 2021, the output value of ginseng exceeded 60 billion, and the market prospect is huge. Due to the high requirements of ginseng for the production environment, continuous cropping directly affects the product quality and output. Various pests and diseases have led to a decline in the output value of ginseng. Therefore, pesticides such as growth promoters, insecticides, and fungicides need to be used for a long time. However, due to the long growth cycle of ginseng, there are a large amount of pesticide residues in both the ginseng body and the soil.

[0003] The "Pharmacopoeia of the People's Republic of China" (2020 Edition) includes total ginsenosides of ginseng stems and leaves and total ginsenosides. The pesticide residues of the ginseng extract (including the crude extract of ginseng stems and leaves) prepared by its preparation method only limit the amounts of hexachlorocyclohexane (total BHC), dichlorodiphenyltrichloroethane (total DDT), and pentachloronitrobenzene (PCNB), but do not limit other pesticide residues. In the prior art, when preparing the crude extract of ginseng stems and leaves and the extract of ginseng roots, water extraction or alcohol extraction is generally used, and then the ginsenosides are enriched by the macroporous resin adsorption method. The total ginsenoside transfer rate of the ginseng extract produced by this method is high, but the amount of enriched pesticide residues is large and difficult to remove. Some stubborn pesticides (such as dimethomorph, propamocarb, etc.) are difficult to meet the removal standards. Summary of the Invention

[0004] The purpose of the present invention is to provide a preparation method of a refined ginseng extract, and the method provided by the present invention can remove various residual pesticides and reduce the pesticide residue content in the ginseng extract.

[0005] In order to achieve the above invention purpose, the present invention provides the following technical solutions:

[0006] The present invention provides a preparation method of a refined ginseng extract, comprising the following steps:

[0007] Mix water, a flocculant with the crude ginseng extract, carry out flocculation treatment, and collect the liquid material after solid-liquid separation;

[0008] Load the liquid material onto a macroporous adsorption resin column, and then elute successively with an alkali solution, water, an acid solution, water, and a first ethanol aqueous solution, and collect the first ethanol aqueous solution eluate;

[0009] Dilute the first ethanol aqueous solution eluate with water, load the resulting diluted solution onto an agricultural residue adsorption resin column, collect the effluent from the column, and then elute with a second ethanol aqueous solution to collect the second ethanol aqueous solution eluate; the resin in the agricultural residue adsorption resin column includes LSA-900D, LX-2000-ss, LX-1600, LSD001, and LX-2;

[0010] Mix the effluent from the column and the second ethanol aqueous solution eluate, and successively perform concentration and drying to obtain a refined ginseng extract.

[0011] Preferably, the flocculant includes ferrous sulfate and calcium hydroxide; the mass ratio of the flocculant to the crude ginseng extract is 1:1 to 10.

[0012] Preferably, the temperature of the flocculation treatment is 60 to 100 °C, and the heat preservation time is 2.5 to 3.5 h.

[0013] Preferably, after the flocculation treatment, a cooling treatment is further included. The temperature of the cooling treatment is 0 to 30 °C, and the time is 12 to 48 h.

[0014] Preferably, the macroporous resin in the macroporous adsorption resin column includes D101 macroporous resin or D101C macroporous resin.

[0015] Preferably, the alkali solution is an aqueous solution of alkali metal hydroxide, and the mass fraction of the alkali solution is 0.1 to 4%.

[0016] Preferably, the acid solution is an aqueous solution of organic acid, and the mass fraction of the acid solution is 0.1 to 4%.

[0017] Preferably, the volume fraction of the first ethanol aqueous solution is 40% to 90%.

[0018] Preferably, the volume fraction of ethanol in the diluted solution is 5% to 40%, and the volume fraction of the second ethanol aqueous solution is 10% to 40%.

[0019] The present invention provides a refined ginseng extract prepared by the preparation method described in the above solution.

[0020] The present invention provides a method for removing agricultural residues from refined ginseng extract, comprising the following steps: mixing water, a flocculant and crude ginseng extract, performing flocculation treatment, separating solid from liquid, and collecting the liquid material; loading the liquid material onto a macroporous adsorption resin column, and then eluting successively with an alkali solution, water, an acid solution, water and a first ethanol aqueous solution, and collecting the first ethanol aqueous solution eluate; diluting the first ethanol aqueous solution eluate with water, loading the obtained diluted solution onto an agricultural residue adsorption resin column, collecting the effluent from the column, and then eluting with a second ethanol aqueous solution, and collecting the second ethanol aqueous solution eluate; the resin in the agricultural residue adsorption resin column comprises LSA-900D, LX-2000-ss, LX-1600, LSD001 and LX-2; mixing the effluent from the column and the second ethanol aqueous solution eluate, and performing concentration and drying successively to obtain refined ginseng extract. In the present invention, the flocculant reacts with the agricultural residue components to generate precipitation, and at the same time, it can ensure that ginsenosides are still in a solution state; furthermore, in the present invention, macroporous resin is used to adsorb ginsenosides, and the flocculant remaining in the solution can also be removed; at the same time, the agricultural residue adsorption resin column composed of a variety of resins in the present invention can fully adsorb various agricultural residue components remaining in the solution. The method provided by the present invention can effectively improve the removal rate of residual pesticides in ginseng extract, has simple operation, and is suitable for large-scale industrial production. Detailed implementation mode

[0021] The present invention provides a method for preparing refined ginseng extract, comprising the following steps:

[0022] Mix water, a flocculant and crude ginseng extract, perform flocculation treatment, separate solid from liquid, and collect the liquid material;

[0023] Load the liquid material onto a macroporous adsorption resin column, and then elute successively with an alkali solution, water, an acid solution, water and a first ethanol aqueous solution, and collect the first ethanol aqueous solution eluate;

[0024] Dilute the first ethanol aqueous solution eluate with water, load the obtained diluted solution onto an agricultural residue adsorption resin column, collect the effluent from the column, and then elute with a second ethanol aqueous solution, and collect the second ethanol aqueous solution eluate; the resin in the agricultural residue adsorption resin column comprises LSA-900D, LX-2000-ss, LX-1600, LSD001 and LX-2;

[0025] Mix the effluent from the column and the second ethanol aqueous solution eluate, and perform concentration and drying successively to obtain refined ginseng extract.

[0026] In the present invention, unless otherwise specified, the raw materials used are all commercially available products well-known to those skilled in the art.

[0027] The present invention mixes water, a flocculant and a crude ginseng extract, performs flocculation treatment, and collects the liquid material after solid-liquid separation. The crude ginseng extract in the present invention is preferably a crude extract of ginseng stems and leaves. In the present invention, the flocculant preferably comprises ferrous sulfate and calcium hydroxide, and the mass ratio of ferrous sulfate to calcium hydroxide is preferably 1:1 to 5, more preferably 1:1; the mass ratio of the flocculant to the crude ginseng extract is preferably 1:1 to 10, further preferably 1:2 to 5, and more preferably 3:10 or 1:2.5. The temperature of the flocculation treatment in the present invention is preferably 60 to 100 °C, further preferably 60 to 65 °C; the heat preservation time is preferably 2.5 to 3.5 h, and specifically can be 2.5 h, 3 h or 3.5 h. The flocculation treatment in the present invention is preferably carried out under stirring conditions. The present invention preferably mixes the crude ginseng extract with water, and then adds the flocculant for flocculation treatment; when adding the flocculant, specifically, ferrous sulfate is added first and stirred for 1 to 30 min, and then calcium hydroxide is added for the flocculation treatment. In the present invention, the mixing of the crude ginseng extract and water is preferably carried out under heating conditions, and the heating temperature is preferably 30 °C to 80 °C, more preferably 60 °C. The present invention preferably mixes the crude ginseng extract with water first, which can convert the crude ginseng extract from a solid state to a liquid state and fully expose the pesticide residues; then adding the flocculant can react the flocculant with some strongly polar anionic pesticides to produce precipitation, and at the same time ensure that ginsenosides are still in a solution state, facilitating the subsequent steps. In the present invention, the flocculation treatment preferably includes a cooling treatment; the temperature of the cooling treatment is preferably 0 to 30 °C, further preferably 2 to 10 °C, and more preferably 4 °C; the time is preferably 12 to 48 h, and specifically can be 12 h, 24 h, 36 h or 48 h. In the present invention, the function of the cooling treatment is to precipitate more non-ginsenoside impurities, facilitate the adsorption in the subsequent process, and prevent the resin column from being blocked. The method of solid-liquid separation in the present invention is preferably filtration.

[0028] After obtaining the liquid material, the present invention samples the liquid material onto a macroporous adsorption resin column, and then elutes it successively with an alkali solution, water, an acid solution, water, a first ethanol aqueous solution, and water, and collects the first ethanol aqueous solution eluate. In the present invention, the macroporous resin in the macroporous adsorption resin column preferably includes D101 macroporous resin or D101C macroporous resin, and more preferably is D101 macroporous resin. The alkali solution in the present invention is preferably an aqueous solution of an alkali metal hydroxide, and the mass fraction of the alkali solution is preferably 0.1-4%; the alkali metal hydroxide in the aqueous solution of the alkali metal hydroxide preferably includes sodium hydroxide or potassium hydroxide, and more preferably is sodium hydroxide; the flow rate of the alkali solution is preferably 0.5-4 BV / h, and more preferably is 2 BV / h. The acid solution in the present invention is preferably an aqueous solution of an organic acid, and the mass fraction of the acid solution is preferably 0.1-4%; the organic acid in the aqueous solution of the organic acid preferably includes one or more of citric acid, oxalic acid, acetic acid, malic acid, and succinic acid, and more preferably is citric acid or oxalic acid; the flow rate of the acid solution is preferably 0.5-4 BV / h, and more preferably is 2 BV / h. In the present invention, when eluting with water, it is preferably eluted until the pH of the obtained water eluate is 6-7. The volume fraction of the first ethanol aqueous solution in the present invention is preferably 40-90%, and more preferably is 60%. The present invention uses a macroporous adsorption resin column to adsorb ginsenosides, and at the same time can also remove the flocculant remaining in the system; the alkali solution can elute the pesticide residues that are weakly bound to the macroporous adsorption resin; the acid solution can not only neutralize the alkali solution, but also elute some non-saponin impurities; eluting with the first ethanol aqueous solution can separate the ginsenosides from the macroporous adsorption resin and achieve enrichment.

[0029] After obtaining the first ethanol aqueous solution eluate, the present invention dilutes the first ethanol aqueous solution eluate with water, samples the resulting dilution on an agricultural residue adsorption resin column, collects the effluent from the column, and then elutes it with a second ethanol aqueous solution to collect the second ethanol aqueous solution eluate. In the present invention, the resin in the agricultural residue adsorption resin column includes LSA-900D, LX-2000-ss, LX-1600, LSD001, and LX-2; the volume ratio of LSA-900D, LX-2000-ss, LX-1600, LSD001, and LX-2 in the agricultural residue adsorption resin column is preferably 1:1:1:1:1. The volume fraction of ethanol in the dilution of the present invention is preferably 5-40%, more preferably 10%. In the present invention, the sample loading rate of the dilution is preferably 0.5-4 BV / h, more preferably 0.5 BV / h. The volume fraction of the second ethanol aqueous solution in the present invention is preferably 10-40%, more preferably 10-30%. In the present invention, the elution rate of the second ethanol aqueous solution is preferably 0.5-4 BV / h, more preferably 0.5 BV / h. The present invention enhances the adsorption strength of the agricultural residue adsorption resin column by adjusting the volume fraction of ethanol in the dilution, and at the same time, the loss of ginsenosides can be reduced. The agricultural residue adsorption resin column of the present invention adopts a combination of multiple resins. Among them, LSA-900D, LX-2000(SS), and LX-1600 are adsorbent resins, LSD001 is a cation resin, and LX-2 is an anion resin; the present invention adopts a combination of multiple resins to fully adsorb various agricultural residue components remaining in the solution.

[0030] After obtaining the effluent from the column and the second ethanol aqueous solution eluate, the present invention mixes the effluent from the column and the second ethanol aqueous solution eluate, and successively performs concentration and drying to obtain a refined ginseng extract. In the present invention, a ginseng extract thick paste is obtained after mixing and concentrating the effluent from the column and the second ethanol aqueous solution eluate, and the solid content of the ginseng extract thick paste is preferably 10-45 wt%, specifically, it can be 10-15 wt% or 30-45 wt%. The present invention has no special limitations on the drying method and conditions, and any drying method well-known to those skilled in the art can be used.

[0031] The present invention preferably performs resin column regeneration treatment on the macroporous adsorption resin column and the agricultural residue adsorption resin column, which can reduce production costs and facilitate industrial large-scale production.

[0032] The present invention also provides a refined ginseng extract prepared by the preparation method described in the above solution.

[0033] In order to further illustrate the present invention, the preparation method of the refined ginseng extract provided by the present invention will be described in detail below with reference to examples, but they should not be construed as limiting the protection scope of the present invention.

[0034] The crude extract of ginseng stem and leaf used in the examples was purchased from Ningbo Xinquan Biotechnology Co., Ltd., with the model number HK230214. The content of ginsenosides in the crude extract of ginseng stem and leaf used in the examples detected by ultraviolet method (UV) was 80%, and the total content of ginsenosides Rg1, Re, and Rd detected by high performance liquid chromatography (HPLC) was 45%.

[0035] The macroporous adsorption resin column used in the examples was purchased from Xi'an BlueSail Biotech Co., Ltd., with the model number D101 macroporous adsorption resin column, the size of Φ500×5000mm, and the height of the resin column was 4000mm; the pesticide residue adsorption resin column used in the examples was purchased from Xi'an BlueSail Biotech Co., Ltd., and the resin and volume ratio therein was LSA-900D: LX-2000-ss: LX-1600: LSD001: LX-2 = 1:1:1:1:1; the size was Φ500×5000mm, and the height of the resin column was 4000mm.

[0036] The reagents used in the examples were all commercially available.

[0037] Example 1

[0038] Add 4000 mL of pure water to a 5000 mL three-necked flask, start stirring, and put in 100 g of the powder of the crude extract of ginseng stem and leaf. Heat to 60 °C to completely disperse the powder of the crude extract of ginseng stem and leaf in water; then add 15 g of ferrous sulfate and stir for 30 min; then add 15 g of calcium hydroxide, continue heating to keep the temperature at 60 °C, and stir and react for 2 h; stop the reaction, cool the three-necked flask to 4 °C, and let it stand at 4 °C for 12 h; filter to remove the precipitate, and then obtain the liquid material through ceramic membrane nanofiltration.

[0039] Load the liquid material onto the D101 macroporous adsorption resin column. After complete adsorption, elute with 4 BV of 2 wt% sodium hydroxide aqueous solution, and the flow rate of the sodium hydroxide aqueous solution is 2 BV / h. Then wash with water until the pH = 6 - 7; then elute with 4 BV of 2 wt% citric acid aqueous solution, and the flow rate of the citric acid aqueous solution is 2 BV / h. Then wash with water until the pH = 6 - 7; then elute with 3 BV of 60 vol% ethanol aqueous solution, and collect the first ethanol aqueous solution eluate.

[0040] Add pure water to the first ethanol aqueous solution eluate to adjust the ethanol concentration to 10 vol% (measured by alcohol meter), and load the obtained dilution onto the pesticide residue adsorption resin column, and the flow rate of the dilution is 0.5 BV / h; then elute the pesticide residue adsorption resin column with 4000 mL of 10 vol% ethanol aqueous solution, and the flow rate of the ethanol aqueous solution is 0.5 BV / h, and collect the second ethanol aqueous solution eluate.

[0041] Mix the effluent from the upper column and the second ethanol aqueous solution eluate for concentration to obtain a thick paste of ginseng extract with a solid content of 10 - 15 wt%. Then, spray-dry the thick paste of ginseng extract to obtain 84 g of off-white ginseng extract, which is the refined ginseng extract.

[0042] Example 2

[0043] Add 2400 kg of pure water to a 3 m 3 alcohol precipitation tank, start stirring, and put in about 60 kg of crude ginseng and ginseng leaf extract powder. Heat to 65 °C to completely disperse the crude ginseng and ginseng leaf extract powder in water. Then add 12 kg of ferrous sulfate and stir for 30 min. Then add 12 kg of calcium hydroxide, continue heating to maintain the temperature at 65 °C, and stir and react for 3 h. Stop the reaction, stir and cool to room temperature, discharge it into a stainless steel barrel, then transfer it to a 4 °C cold storage, and let it stand at 4 °C for 48 h. First, remove the precipitate through a 2 m² stainless steel plate and frame filter, and then subject the filtrate to ceramic membrane nanofiltration to obtain a liquid material.

[0044] Load the liquid material onto a D101 macroporous adsorption resin column. After complete adsorption, elute with 3000 kg of 2 wt% sodium hydroxide aqueous solution at a flow rate of 2 BV / h for the sodium hydroxide aqueous solution, and then wash with water until the pH = 6 - 7. Add 3000 kg of 2 wt% citric acid aqueous solution for elution at a flow rate of 2 BV / h for the citric acid aqueous solution, and then wash with water until the pH = 6 - 7. Then elute with 2400 L of 60 vol% ethanol aqueous solution and collect the first ethanol aqueous solution eluate.

[0045] Add pure water to the first ethanol aqueous solution eluate to adjust the ethanol concentration to 10 vol% (measured with an alcohol meter). Load the resulting dilution onto a pesticide residue adsorption resin column at a flow rate of 0.5 BV / h for the dilution, and then elute the pesticide residue adsorption resin column with 2000 L of 10 vol% ethanol aqueous solution at a flow rate of 0.5 BV / h for the ethanol aqueous solution, and collect the second ethanol aqueous solution eluate.

[0046] Mix the effluent from the upper column and the second ethanol aqueous solution eluate for concentration to obtain a thick paste of ginseng extract with a solid content of 30 - 45 wt%. Then, spray-dry the thick paste of ginseng extract to obtain 55.8 kg of off-white ginseng extract, which is the refined ginseng extract.

[0047] Resin column regeneration treatment: Wash the resin successively with 4 BV of 4 wt% sodium hydroxide aqueous solution, pure water (RO membrane secondary reverse osmosis water) until the pH = 7 - 8, 4 BV of 4 wt% hydrochloric acid solution, pure water until the pH = 6 - 7, 4 BV of 90 vol% ethanol aqueous solution for elution, and then wash with pure water until there is no alcohol smell.

[0048] The pesticide residues in the crude extract of ginseng stems and leaves according to the present invention are shown in Table 1.

[0049] Table 1 Pesticide residues in the crude extract of ginseng stems and leaves

[0050]

[0051]

[0052] The pesticide residues in the refined ginseng extract prepared in Example 2 of the present invention are shown in Table 2.

[0053] Table 2 Pesticide residues in the refined ginseng extract prepared in Example 2

[0054]

[0055]

[0056] As can be seen from Table 1 and Table 2, by treating the crude extract of ginseng stems and leaves according to the method provided in this application to obtain a refined ginseng extract, the removal rates of more than twenty pesticide residues such as fluxapyroxad, azoxystrobin, boscalid, difenoconazole, dimethomorph, etc. in the crude extract of ginseng stems and leaves reach 100%; the removal rates of carbendazim, carbendazim and benomyl reach 99.7%, and the removal rate of procymidone reaches 92.9%. Thus, it can be seen that the method provided in this application can effectively improve the removal rate of residual pesticides in the crude extract of ginseng stems and leaves.

[0057] Although the above embodiments have described the present invention in detail, they are only a part of the embodiments of the present invention, rather than all embodiments. People can also obtain other embodiments based on these embodiments without creative efforts, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A method for preparing a refined ginseng extract, comprising the following steps: Mix water, a flocculant and a crude ginseng extract, carry out flocculation treatment, and collect the liquid material after solid-liquid separation; the flocculant is ferrous sulfate and calcium hydroxide, and the mass ratio of ferrous sulfate to calcium hydroxide is 1:1-5; the addition of the flocculant is to first add ferrous sulfate and then add calcium hydroxide; the temperature of the flocculation treatment is 60-100 °C; the crude ginseng extract is a crude extract of ginseng stems and leaves; after the flocculation treatment, a cooling treatment is also included; the temperature of the cooling treatment is 0-30 °C, and the time is 12-48 h; Load the liquid material onto a macroporous adsorption resin column, and then elute successively with an alkali solution, water, an acid solution, water and a first ethanol aqueous solution, and collect the first ethanol aqueous solution eluate; the alkali solution is an aqueous solution of an alkali metal hydroxide, and the mass fraction of the alkali solution is 0.1-4%; the acid solution is an aqueous solution of an organic acid, and the mass fraction of the acid solution is 0.1-4%; the volume fraction of the first ethanol aqueous solution is 40%-90%; Dilute the first ethanol aqueous solution eluate with water, load the obtained diluted solution onto a pesticide adsorption resin column, collect the effluent from the column, and then elute with a second ethanol aqueous solution, and collect the second ethanol aqueous solution eluate; the resin in the pesticide adsorption resin column is LSA-900D, LX-2000-ss, LX-1600, LSD001 and LX-2; the volume ratio of LSA-900D, LX-2000-ss, LX-1600, LSD001 and LX-2 in the pesticide adsorption resin column is 1:1:1:1:1; the volume fraction of ethanol in the diluted solution is 5-40%, and the volume fraction of the second ethanol aqueous solution is 10-40%; Mix the effluent from the column and the second ethanol aqueous solution eluate, and carry out concentration and drying in sequence to obtain a refined ginseng extract.

2. The preparation method according to claim 1, wherein, The mass ratio of the flocculant to the crude ginseng extract is 1:1-10.

3. The preparation method according to claim 1 or 2, characterized in that, The heat preservation time of the flocculation treatment is 2.5-3.5 h.

4. The preparation method according to claim 1, characterized in that, The macroporous resin in the macroporous adsorption resin column includes D101 macroporous resin or D101C macroporous resin.

5. A refined ginseng extract prepared by the preparation method according to any one of claims 1 to 4.

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