1. Ginsenoside Rg₁The preparation method of the molecularly imprinted polymeric material is characterized by comprising the following steps:

1) mixing template molecules and functional monomers, dissolving the mixture in a pore-forming agent, uniformly mixing the mixture by ultrasonic waves, adding an effective amount of a cross-linking agent, quickly stirring the mixture to form a suspension, adding an initiator, introducing nitrogen for at least 30 min to remove dissolved oxygen, and sealing the mixture;

2) under the protection of nitrogen, the molecularly imprinted polymer is initiated by heat and is placed in a magnetic stirrer with a heating plate for polymerization reaction;

3) after the polymerization reaction is finished, eluting the obtained molecularly imprinted polymer with a mixed solution of methanol, acetic acid and water, carrying out high performance liquid detection on an eluent until no template substance is detected, washing the eluent with methanol to be neutral, and then carrying out vacuum drying at 40-60 ℃ for 10-24 hours to prepare ginsenoside Rg₁A molecularly imprinted polymer.

2. The method of claim 1, wherein the functional monomer is 4-vinylphenylboronic acid, 3-acrylamidophenylboronic acid, triaminophenylboronic acid.

3. The method of claim 1, wherein the ginsenoside Rg is administered₁The molar ratio of the functional monomer to the functional monomer is 1: 4-15, preferably 1: 10-15.

4. The method of claim 1, wherein the porogen is absolute ethanol, absolute methanol, isopropanol and acetonitrile, preferably absolute ethanol and absolute methanol.

5. The method of claim 1, wherein the volume dosage of the porogen is 5-15 mL.

6. The method of claim 1, wherein the cross-linking agent is ethylene glycol dimethacrylate, trimethylolpropane trimethacrylate, N, N-methylenebisacrylamide, and the amount of the cross-linking agent is 0.3 to 0.9 mmol.

7. The method of claim 1, wherein the initiator is azobisisobutyronitrile and azobisisoheptonitrile, preferably azobisisobutyronitrile, and the amount of the initiator is 0.4-1.2 mL (15 mg/mL).

8. The method according to claim 1, wherein the magnetic stirrer rotates at a speed of 400r/min to 1500r/min, preferably 1000 r/min to 1500 r/min.

9. The process according to claim 1, wherein the polymerization temperature is 40 to 60 ℃.

10. The process according to claim 1, wherein the polymerization time is 3 to 24 hours, preferably 12 to 24 hours.

11. The process according to claim 1, wherein the elution is carried out by sonication, magnetic stirring and shaking with a constant temperature shaker, preferably with a constant temperature shaker.

12. The method according to claim 1, wherein the eluent is acidic methanol or ethanol water solution, the acidity is adjusted by formic acid or acetic acid, and the volume ratio of the eluent is methanol or ethanol (43-92): formic acid or acetic acid (8-12): water (0-45), preferably, the volume ratio of methanol-acetic acid-water is 60-65: 8-10: 25-32.

13. Ginsenoside Rg₁The molecularly imprinted polymer is characterized in that the molecularly imprinted polymeric material prepared by the preparation method of claims 1 to 12 is used for adsorbing and separating ginsenoside Rg₁The use of (1).

14. The use of the molecularly imprinted polymer according to claim 13, wherein the ginsenoside solution is prepared by mixing ginsenoside Rg1 with a mass percentage of 90.0% calculated by a standard curve of ginsenoside Rg1 and an aqueous ethanol solution with a volume percentage of 30-50%; the constant temperature oscillation temperature is 20-40 ℃, the rotation speed is 150-200 rpm, and the oscillation time is 2-24 h.

Background

The Ginseng radix is Panax ginseng C.A. Meyer of AraliaceaePanax ginsengThe dried roots and the rhizomes of C.A. Mey are traditional and rare traditional Chinese medicines in China, are originally recorded in the Shen nong's herbal Jing which is the first herbal monograph in China, and have the effects of tonifying qi, generating blood, strengthening body resistance, eliminating pathogenic factors and the like. Meanwhile, ginseng is also the main medicinal flavor of ginseng, poria, bighead atractylodes rhizome powder, kaiki powder and other classical famous prescriptions, and plays an important role in preventing and treating diseases. The modern pharmacological action research shows that ginseng plays a role in various systems of the human body, such as the central nervous system, the cardiovascular system, the respiratory system, the blood and hematopoietic system, the endocrine system, the reproductive system, the immune system and the like. Ginsenoside is the main active component in ginseng, and has multiple functions of resisting aging, resisting oxidation, improving immunity, enhancing memory, etc.

The ginsenoside components are in a large quantity, about 50 ginsenoside monomers are known clearly, and the content of the ginsenoside monomers in ginseng is about 4 percent. Ginsenoside Rg₁Is one of the main active components of ginsenoside, belongs to protopanaxatriol saponin, has multiple activities of cardiovascular protection, nervous system protection, anti-aging, anti-inflammation, anti-tumor and the like, and is one of the main index components for measuring the content of ginseng. At present, the content determination of ginsenoside mainly adopts high performance liquid chromatography, and the structures of various components of ginsenoside are similar, and the difference of physicochemical properties is small, so that the chromatographic behaviors of monomers are close, therefore, the separation difficulty in the content determination process is large, and the accurate detection is difficult. Moreover, the content of ginsenoside is low, and the separation and detection of ginsenoside by chemical components of other medicinal ingredients of the traditional Chinese medicine compound have great interference, so that the difficulty of accurately determining the content of ginsenoside is increased.

Molecularly Imprinted Polymers (MIPs) refer to the method of using a target molecule as a template, combining a functional monomer with a complementary structure with the template molecule in a covalent or non-covalent manner, adding a cross-linking agent for polymerization reaction, and after the reaction is finished, polymerizing the template moleculeThe high cross-linked polymer with fixed cavity size and shape and definite arrangement functional group is formed by elution. The prepared material has extremely high selectivity and excellent molecular recognition performance, and is quickly and widely applied to aspects of solid phase extraction, artificial enzymology, chiral resolution, biosensors, asymmetric catalysis and the like. Therefore, the establishment of a method for preparing ginsenoside Rg₁The preparation method of the molecular imprinting polymer utilizes the prepared molecular imprinting polymer to be applied to the ginsenoside Rg in food, medicine and biological samples₁In the pretreatment method for content determination, the ginsenoside Rg in the sample is improved₁The extraction rate and the detection accuracy rate of the method eliminate the interference and influence of other components on the method.

Disclosure of Invention

The invention provides ginsenoside Rg₁A molecularly imprinted polymer is prepared from ginsenoside Rg₁Adding functional monomer and cross-linking agent as template, respectively, thermally initiating, and precipitating to polymerize to obtain ginsenoside Rg₁Ginsenoside Rg with specific selectivity₁Molecularly imprinted polymer for improving ginsenoside Rg₁The extraction rate and the adsorption rate of the traditional Chinese medicine composition can eliminate the interference and influence of other components in single and compound traditional Chinese medicines.

One aspect of the invention provides ginsenoside Rg₁The preparation method of the molecularly imprinted polymeric material comprises the following steps:

(1) mixing template molecules and functional monomers, dissolving the mixture in a pore-forming agent, uniformly mixing the mixture by ultrasonic waves, adding an effective amount of a cross-linking agent, quickly stirring the mixture to form a suspension, adding an initiator, introducing nitrogen for at least 30 min to remove dissolved oxygen, and sealing the mixture;

(2) under the protection of nitrogen, the molecularly imprinted polymer is initiated by heat and is placed in a magnetic stirrer with a heating plate for polymerization reaction;

(3) after the polymerization reaction is finished, eluting the obtained molecularly imprinted polymer with a mixed solution of methanol, acetic acid and water, carrying out high performance liquid detection on an eluent until no template substance is detected, washing the eluted liquid to be neutral with methanol, and then carrying out vacuum drying at 50 ℃ for 12-24 hours to obtain the ginsengSaponin Rg₁A molecularly imprinted polymer.

Further, the functional monomer in the step (1) is one of 4-vinylphenylboronic acid, 3-acrylamidophenylboronic acid and triaminophenylboronic acid. Ginsenoside Rg₁The molar ratio of the functional monomer to the functional monomer is 1 (4-15), and preferably 1 (10-15).

Further, the pore-foaming agent in the step (1) is one of absolute ethyl alcohol, absolute methyl alcohol, acetonitrile and isopropanol. The volume consumption of the pore-foaming agent is 5-15 mL.

Further, the crosslinking agent in the step (1) is one of ethylene glycol dimethacrylate, trimethylolpropane trimethacrylate and N, N-methylene bisacrylamide. The dosage of the cross-linking agent is 0.3-0.9 mmol.

Further, the initiator in the step (1) is one of azobisisobutyronitrile and azobisisoheptonitrile, and the dosage of the initiator is 0.4-1.2 mL (15 mg/mL).

Further, the rotating speed of the magnetic stirrer in the step (1) is 400 r/min-1500 r/min, preferably 1000 r/min-1500 r/min.

Further, the polymerization temperature in the step (1) is 40-60 ℃.

Further, in the step (1), the polymerization reaction time is 3-24 hours, preferably 12-24 hours.

Further, the elution mode in the step (1) is ultrasonic, magnetic stirring and constant temperature shaking table oscillation, and preferably constant temperature shaking table oscillation.

Further, the eluent in the step (1) is acidic methanol or ethanol water solution, formic acid or acetic acid is adopted for acidity adjustment, and the volume ratio of the eluent is methanol or ethanol (43-92): formic acid or acetic acid (8-12): water (0-45), preferably, the volume ratio of methanol-acetic acid-water is 60-65: 8-10: 25-32.

Ginsenoside Rg₁The molecularly imprinted polymer is characterized in that the molecularly imprinted polymeric material prepared by the preparation method of claims 1 to 12 is used for adsorbing and separating ginsenoside Rg₁The use of (1). Characterized in that the ginsenoside solution is prepared by ginsenoside Rg₁Ginsenoside Rg with the mass percentage of 90.0 percent calculated by a standard curve₁Mixing the mixture with 30 to 50 volume percent of ethanol water solution to prepare the composite material; the constant temperature oscillation temperature is 20-40 ℃, the rotation speed is 150-200 rpm, and the oscillation time is 2-24 h.

The invention also provides the application of the molecular imprinting polymer material in separating and enriching the ginsenoside Rg in the traditional Chinese medicine compound Kaixin powder and the single traditional Chinese medicine ginseng₁The application of (2), comprising the following steps: taking the above molecularly imprinted polymer as solid phase extraction adsorbent, loading the extract of the Chinese medicinal composition KAIXIN powder into column, eluting with 30% methanol solution, and eluting with 70% methanol solution to obtain eluate containing ginsenoside Rg₁。

The invention has the advantages and effects that: the invention relates to ginsenoside Rg₁Contains a plurality of glycosyl characteristics, preferably selects functional monomers, constructs the ginsenoside Rg based on boric acid group₁A reversible covalent molecularly imprinted polymer. The polymer overcomes the influence that the traditional non-covalent imprinted polymer is limited by a solvent, and the preparation and the identification of the polymer can be carried out in a strong polar solvent such as alcohol or water. The invention adopts the constant temperature shaking table to elute the template molecules by oscillation, and compared with the elution modes of Soxhlet extraction, ultrasonic elution, magnetic stirring and the like reported in research, the constant temperature shaking table has better elution effect by oscillation, and only oscillates and elutes for 2 hours, and the elution rate of the template molecules reaches 70.58 percent.

Drawings

FIG. 1, a graph for abstract, ginsenoside Rg₁Schematic diagram of preparation of imprinted polymeric materials.

FIG. 2, ginsenoside Rg prepared in example 1₁Fourier infrared spectra of imprinted polymeric materials.

FIG. 3, ginsenoside Rg prepared in example 1₁Scanning electron microscopy of imprinted polymeric materials.

Fig. 4, graph of adsorption kinetics of molecularly imprinted polymeric Materials (MIPs) and non-imprinted polymeric materials (NIPs) to ginsenoside Rg1 in example 1.

FIG. 5, example1 molecular imprinting polymer Materials (MIPs) and non-imprinting polymer materials (NIPs) of molecular imprinting polymer Materials (MIPs) and ginsenoside Rg₁Adsorption isotherm diagram of (1).

FIG. 6 shows that molecular imprinted polymeric Materials (MIPs) and non-imprinted polymeric materials (NIPs) in example 1 have effect on ginsenoside Rg₁、Re、Rb₁And a comparison graph of the selective adsorption effect of rutin and quercetin.

Detailed Description

The chemical reagents used: 4-vinylphenylboronic acid (4-VPBA), 3-acrylamidophenylboronic acid, triaminophenylboronic acid (APBA); ethylene Glycol Dimethacrylate (EGDMA), Diethylbenzene (DVB), trimethylolpropane Trimethacrylate (TRIM), N-Methylenebisacrylamide (MBAA); azobisisobutyronitrile (AIBN), Azobisisoheptonitrile (ABVN). : absolute ethanol, absolute methanol and isopropanol.

Example 1

(1) Mixing 45.75 mg (0.05 mmol) of template molecule ginsenoside Rg₁75.53 mg (0.5 mmol) of functional monomer 4-VPBA is added into 10 mL of absolute ethyl alcohol and dissolved by ultrasonic; adding 132 μ L (0.7 mmol) crosslinking agent EGDMA, and stirring at high speed to form suspension; after adding 0.8 mL of AIBN initiator (15 mg/mL), nitrogen purge for at least 30 min to completely remove dissolved oxygen, seal;

(2) under the protection of nitrogen, the molecularly imprinted polymer is initiated by heat and is placed in a magnetic stirrer with a heating plate for polymerization: the polymerization reaction is carried out for 12 hours at the temperature of 60 ℃ and at the speed of 1500 r/min.

(3) After the polymerization reaction is finished, suction filtration is carried out, and the obtained polymer is dried in a vacuum drying oven for 12 hours at the temperature of 60 ℃. And then, putting the polymer in a constant-temperature culture oscillator by taking methanol-acetic acid-water (63: 10: 27, V/V) as an eluting solvent, shaking to elute the template molecules until the template molecules cannot be detected by the high-performance liquid phase of the eluent, washing the eluent to be neutral by using methanol, and drying to obtain the MIPs. The yield thereof was found to be 85%.

(4) Except that the ginsenoside Rg is not added in the synthesis process₁Except that, the rest steps are the same as the preparation steps of the imprinted polymer, and non-imprinted polymeric materials (NIPs) are obtained.

(5) And (3) performing character characterization on the prepared molecularly imprinted polymer. FIG. 2 shows polymerized ginsenoside Rg without a release plate₁Fourier infrared spectrogram of the molecularly imprinted polymer, infrared spectrogram analysis: 3483cm-1 as template molecule Rg₁The association-OH stretching vibration absorption peak is 1604cm-1, 1556cm-1 and 1455cm-1, and is the stretching vibration absorption peak of the benzene ring skeleton of the functional monomer 4-VPBA; 1728cm-1 is the stretching vibration of the crosslinking agent EGDMA ester group C = O. Infrared spectrogram analysis shows that the template molecule ginsenoside Rg1, the functional monomer 4-VPBA and the cross-linking agent EGDMA are successfully cross-linked and polymerized on the polymer in the polymerization process. FIG. 3 shows ginsenoside Rg prepared in this example₁The scanning electron microscope image of the molecularly imprinted polymeric material shows that the polymer microspheres have uniform particle size and good monodispersity.

Example 2

The ginsenoside Rg of the invention₁Adsorption separation of ginsenoside Rg from imprinted polymeric material₁The application of (1):

(1) ginsenoside Rg under different oscillation adsorption time conditions₁Comprises the following steps: accurately weighing 5mg of MIPs and NIPs in example 1 respectively in a 15mL centrifuge tube, and adding 1 mL of 0.5 mg/mL ginsenoside Rg₁Placing the ethanol solution in a constant temperature culture oscillator, oscillating and adsorbing at 20 deg.C, centrifuging for 2, 4, 6, 8, 12, 18 and 24h respectively to obtain supernatant, and analyzing Rg in the adsorption solution by high performance liquid chromatography₁The concentration change of (2) is calculated according to the mass concentration difference before and after adsorption₁The change of the adsorption amount of (D) Q (. mu.g/mg) with time.

(2) The results are shown in FIG. 4, which shows that the molecular imprinted polymeric Materials (MIPs) and the non-imprinted polymeric materials (NIPs) in this example are applied to ginsenoside Rg₁Graph of adsorption kinetics of (a). As can be seen from the graph, the adsorption amounts of MIPs and NIPs are gradually increased along with the time, and the adsorption equilibrium is achieved in 12 h. The adsorption quantity of the MIPs in adsorption equilibrium is 31.90 mu g/mg, the adsorption quantity of the NIPs is 14.58 mu g/mg, and the equilibrium adsorption quantity of the MIPs is 2.2 times of the NIPs, which shows that the MIPs has obvious imprinting effect.

Example 3 substrate concentration vs. ginsenosideRg₁Effect of the adsorption Capacity of imprinted polymeric materials

Precisely weighing several parts of 5mg each of MIPs and NIPs in example 1 in a 15mL centrifuge tube, and sequentially adding 1 mL of ginsenoside Rg with the concentration of 0.5 mg/mL, 1 mg/mL, 1.5 mg/mL, 2.0 mg/mL and 2.5 mg/mL₁Placing the ethanol solution in a constant temperature culture oscillator, oscillating and adsorbing at 20 deg.C for 12h, centrifuging to obtain supernatant, and analyzing Rg in the adsorption solution according to high performance liquid chromatography detection under Ginseng radix item of China pharmacopoeia 2020 edition₁The concentration change of (2) is calculated according to the mass concentration difference before and after adsorption₁The adsorption amount of (1) Q (. mu.g/mg). The results are shown in FIG. 5, which shows that the molecular imprinted polymeric Materials (MIPs) and the non-imprinted polymeric materials (NIPs) in this example are applied to ginsenoside Rg₁Adsorption isotherm diagram of (1). As can be seen from the figure, in a certain concentration range, the ginsenoside Rg in the absorption liquid is accompanied₁Increase in concentration, MIPs and NIPs to Rg₁The amount of adsorption of (b) also gradually increases. When Rg is in the adsorption liquid₁When the initial concentration is increased to 2.0 mg/mL, the MIPs and the NIPs reach the adsorption saturation, the saturated adsorption capacity of the MIPs and the NIPs is 62.22 mu g/mg and 29.70 mu g/mg respectively, the maximum saturated adsorption capacity of the MIPs is 2.1 times of that of the NIPs, and the result shows that the MIPs can realize the ginsenoside Rg to the ginsenoside Rg₁The adsorption of (a) is a selective adsorption with specific recognition sites, unlike the pure physical adsorption of NIPs. The specific molecular recognition effect depends on the geometrical selectivity of MIPs imprinted pores and the reversible covalent bond between the functional monomer and the template molecule.

Example 4 ginsenoside Rg₁Molecular engram polymeric material for ginsenoside Rg₁Adsorption selectivity of

(1) Several portions of 5mg each of MIPs and NIPs in example 1 were precisely weighed and divided into 2 groups. Adding 1 mL 2.0 mg/mL ginsenoside Rg into the first group of MIPs and NIPs respectively₁、Re、Rb₁An ethanol mixed solution of rutin and quercetin; adding substrate solution (ginsenoside Rg) with different concentrations into the second group of MIPs and NIPs respectively₁ 0.5 mg/mL、Re 1.5 mg/mL、Rb₁2.0 mg/mL, rutin 2.5 mg/mL, quercetin 2.5 mg/mL). Placing in a constant temperature culture oscillator, oscillating at 20 deg.C for 12 hr for adsorption, centrifuging to obtain supernatant, and detecting and analyzing each adsorption with high performance liquid chromatographyThe concentration of the solution changes. Specific adsorption capacity of MIPs was evaluated as adsorption quantity Q (μ g/mg). The results are shown in FIG. 6, which shows that the molecular imprinted polymeric Materials (MIPs) and the non-imprinted polymeric materials (NIPs) in this example are applied to ginsenoside Rg₁、Re、Rb₁And a comparison graph of the selective adsorption effect of rutin and quercetin. As can be seen from A diagram, at the same concentration level, MIPs can treat the template molecule ginsenoside Rg₁With preferential selectivity for its structural analogs Re and Rb₁Is significantly less selective than the template molecule. This is due to the fact that MIPs have the same Rg₁Matched specific binding sites capable of selectively adsorbing template molecules Rg₁(ii) a And ginsenoside Re and Rb₁The molecular structure of the compound contains ginsenoside Rg₁The same parent nucleus, except that the number and the connection position of glycosyl groups contained in the structure are different, leads the glycosyl groups not to enter the imprinting holes perfectly in the molecular recognition and re-adsorption process of the MIPs, so that the MIPs have lower adsorption to the glycosyl groups than the template molecules at the same concentration level. In addition, MIPs also have a certain adsorption to rutin, which is a flavonoid compound, because rutin contains 2 glycosyl groups in the molecular structure, but the parent nucleus structure of the rutin and ginsenoside Rg₁The difference is too large, so that the adsorption quantity of the MIPs to the MIPs is lower than that of the ginsenoside; MIPs are not selective for the flavonoid quercetin. Because quercetin does not contain glycosyl, cis-diol structure does not exist in the structure, so that MIPs cannot combine with quercetin, and the adsorption quantity of the quercetin is less than that of NIPs. The NIPs are only physically adsorbed, so that ginsenoside Rg is added₁、Re、Rb₁And the adsorption amounts of rutin and quercetin are not greatly different. The results show that the template molecule ginsenoside Rg₁The sugar units in the structure covalently interact with the functional monomer 4-VPBA and ultimately create recognition sites in the cavities of the polymer. As shown in FIG. B, MIPs have adsorption selectivity for both template molecules and their structural analogs, and are concentration dependent. Within a certain range, the higher the substrate concentration, the higher the adsorption capacity. There was still no adsorption selectivity for elevated concentrations of quercetin that did not contain sugar groups.

Example 5 ginsenoside Rg of the invention₁Separation and enrichment of imprinted polymeric material in traditional Chinese medicine compoundGinsenoside Rg in Kaixin powder₁And structural analogs Re and Rb thereof₁The application of (1):

(1) preparing an extracting solution of the Kaixuan powder: precisely weighing 2.5 g of pistachio powder in a conical flask, adding 25 mL of 70% methanol, weighing, ultrasonically extracting for 0.5 h, cooling, weighing again, complementing the lost weight with 70% methanol, shaking up, filtering, taking the subsequent filtrate, evaporating to dryness in an evaporating dish, adding water to dissolve the residue, fixing the volume to a 5mL measuring flask, and shaking up to obtain the finished product.

(2) Preparation of a molecularly imprinted solid phase extraction column (MIP-SPE): weighing about 25 mg each of MIPs and NIPs in example 1, respectively filling into empty SPE small columns, and fixing the upper end and the lower end by using sieve plates. The MIP-SPE cartridge was activated sequentially with 1 mL of methanol, 1 mL of 50% methanol, and 2mL of ultrapure water for use.

(3) Separating and purifying saponin components in the extract of the kaixiong powder by MIP-SPE combined method: mu.L of Kaixuan extract was slowly passed through the MIP-SPE cartridge at a flow rate to load the sample, then the cartridge was rinsed with 30% methanol in portions (1 mL. times.3) to eliminate non-selective adsorption, and the eluate was collected. Finally, the target was eluted in portions (1 mL. times.8) with 8 mL of 70% methanol-glacial acetic acid (9: 1, V/V) solution, the eluates were collected, evaporated to dryness in an evaporating dish, redissolved in methanol and taken up in a 5mL volumetric flask. And (3) taking the extract of the kaixuan powder, the sample effluent, the eluent and the eluent to respectively carry out high performance liquid analysis. The results of the measurements are shown in Table 1 below.

]Table 1: MIP-SPE method for extracting and separating ginsenoside Rg from Kaixing powder₁Re and Rb₁(n=3)

As can be seen from Table 1, 25 mg of MIPs adsorbed ginsenoside Rg respectively₁Re and Rb₁59.25, 457.12 and 907.33 μ g, respectively account for the ginsenoside Rg in the sample solution₁Re and Rb₁58%, 61% and 62% of the total amount. Finally, eluting by using 70% methanol-glacial acetic acid (9: 1, V/V) solution as an eluent in batches, and eluting 84% -90% of target components. After NIPs sample loading and adsorption, the ginseng soap in effluent liquidGlycoside Rg₁Re and Rb₁The peak area of the elution reagent is almost unchanged, a large amount of target components appear in 30% methanol leacheate, and the calculated recovery rate of the target components is only 7% -12%. Indicating that no specific binding sites are present in the NIPs, but pure physisorption. Shows that the synthesized MIPs have the template molecule ginsenoside Rg₁Ginsenoside Re and Rb as structural analogs thereof₁Has higher affinity and selectivity, and MIP-SPE is an effective method for extracting ginsenoside in a complex matrix sample.

Example 6 ginsenoside Rg of the invention₁The preparation method of the molecularly imprinted polymeric material comprises the following steps:

(1) mixing 45.75 mg (0.05 mmol) of template molecule ginsenoside Rg₁113.29 mg (0.75 mmol) of functional monomer 4-VPBA is added into 15mL of anhydrous methanol and dissolved by ultrasonic; adding 170 μ L (0.9 mmol) crosslinking agent EGDMA, and stirring at high speed to form suspension; after adding 1.2mL of AIBN initiator (15 mg/mL), nitrogen was purged for at least 30 min to completely remove dissolved oxygen and sealed;

(2) under the protection of nitrogen, the molecularly imprinted polymer is initiated by heat and is placed in a magnetic stirrer with a heating plate for polymerization: polymerization reaction is carried out for 24 hours at 50 ℃ and 1000 r/min.

(3) After the polymerization reaction is finished, suction filtration is carried out, and the obtained polymer is dried in a vacuum drying oven for 24 hours at the temperature of 40 ℃. And then, putting the polymer in a constant-temperature culture oscillator by using ethanol-formic acid-water (65: 10:25, V/V) as an eluting solvent, shaking to elute the template molecules until the template molecules cannot be detected by the high-performance liquid phase of the eluent, washing the eluent to be neutral by using methanol, and drying to obtain the MIPs. The yield thereof was found to be 75%.

Example 7 ginsenoside Rg of the invention₁The preparation method of the molecularly imprinted polymeric material comprises the following steps:

(1) mixing 45.75 mg (0.05 mmol) of template molecule ginsenoside Rg₁90.64 mg (0.6 mmol) of functional monomer 4-VPBA is added into 10 mL of absolute ethyl alcohol and dissolved by ultrasonic; adding 132 μ L (0.7 mmol) crosslinking agent EGDMA, and stirring at high speed to form suspension; after addition of 0.8 mL of AIBN initiator (15 mg/mL), it was purged with nitrogen for at least 30 min to remove completelyDissolving oxygen, and sealing;

(2) under the protection of nitrogen, the molecularly imprinted polymer is initiated by heat and is placed in a magnetic stirrer with a heating plate for polymerization: the polymerization reaction is carried out for 12 hours at the temperature of 60 ℃ and at the speed of 1500 r/min.

(3) After the polymerization reaction is finished, suction filtration is carried out, and the obtained polymer is dried in a vacuum drying oven for 12 hours at the temperature of 60 ℃. And then, putting the polymer in a constant-temperature culture oscillator by taking methanol-acetic acid-water (63: 10: 27, V/V) as an eluting solvent, shaking to elute the template molecules until the template molecules cannot be detected by the high-performance liquid phase of the eluent, washing the eluent to be neutral by using methanol, and drying to obtain the MIPs. The yield thereof was found to be 78%.

Example 8 ginsenoside Rg of the invention₁The preparation method of the molecularly imprinted polymeric material comprises the following steps:

(1) mixing 45.75 mg (0.05 mmol) of template molecule ginsenoside Rg₁38.198 mg (0.2 mmol) of functional monomer 3-acrylamidophenylboronic acid is added into 5mL of acetonitrile, and the mixture is dissolved by ultrasonic wave; adding 56 μ L (0.3 mmol) crosslinking agent trimethylolpropane trimethacrylate, and stirring at high speed to form suspension; after adding 0.4mL of azobisisoheptonitrile (15 mg/mL) as an initiator, nitrogen was purged for at least 30 min to completely remove dissolved oxygen and sealed;

(2) under the protection of nitrogen, the molecularly imprinted polymer is initiated by heat and is placed in a magnetic stirrer with a heating plate for polymerization: polymerization is carried out for 24 hours at 40 ℃ and 400 r/min.

(3) After the polymerization reaction is finished, suction filtration is carried out, and the obtained polymer is dried in a vacuum drying oven for 10 hours at the temperature of 60 ℃. And then, putting the polymer in a constant-temperature culture oscillator by taking methanol-formic acid-water (43: 12: 45, V/V) as an eluting solvent, shaking to elute the template molecules until the template molecules cannot be detected by the high-performance liquid phase of the eluent, washing the eluent to be neutral by using methanol, and drying to obtain the MIPs. The yield thereof was found to be 45%.

]Example 9 ginsenoside Rg of the invention₁The preparation method of the molecularly imprinted polymeric material comprises the following steps:

(1) mixing 45.75 mg (0.05 mmol) of template molecule ginsenoside Rg₁102.71 mg (0.75 mmol) of the functionAdding a monomer triaminophenylboronic acid into 15mL of isopropanol, and dissolving by ultrasonic treatment; adding 114 μ L (0.6 mmol) of cross-linking agent N, N-methylene bisacrylamide, and stirring at high speed to form a suspension; after 1.0mL of azobisisoheptonitrile (15 mg/mL) as an initiator was added, nitrogen was purged for at least 30 min to completely remove dissolved oxygen and sealed;

(2) under the protection of nitrogen, the molecularly imprinted polymer is initiated by heat and is placed in a magnetic stirrer with a heating plate for polymerization: polymerization reaction is carried out for 16h at 50 ℃ and 1000 r/min.

(3) After the polymerization reaction is finished, suction filtration is carried out, and the obtained polymer is dried in a vacuum drying oven for 24 hours at the temperature of 50 ℃. And then, putting the polymer in a constant-temperature culture oscillator by taking methanol-acetic acid-water (60: 8: 32, V/V) as an eluting solvent, shaking to elute the template molecules until the template molecules cannot be detected by the high-performance liquid phase of the eluent, washing the eluent to be neutral by using methanol, and drying to obtain the MIPs. The yield thereof was found to be 58.5%.

]Example 10 ginsenoside Rg of the invention₁The preparation method of the molecularly imprinted polymeric material comprises the following steps:

(1) 45.75 mg (0.05 mmol) of template molecule ginsenoside Rg1 and 113.30 mg (0.75 mmol) of functional monomer 4-VPBA are added into 10 mL of absolute ethyl alcohol and dissolved by ultrasonic; adding 132 μ L (0.7 mmol) crosslinking agent EGDMA, and stirring at high speed to form suspension; after adding 0.8 mL of AIBN initiator (15 mg/mL), nitrogen purge for at least 30 min to completely remove dissolved oxygen, seal;

(2) under the protection of nitrogen, the molecularly imprinted polymer is initiated by heat and is placed in a magnetic stirrer with a heating plate for polymerization: the polymerization reaction is carried out for 12 hours at the temperature of 60 ℃ and at the speed of 1500 r/min.

(3) After the polymerization reaction is finished, suction filtration is carried out, and the obtained polymer is dried in a vacuum drying oven for 12 hours at the temperature of 60 ℃. And then, putting the polymer in a constant-temperature culture oscillator by taking methanol-acetic acid-water (63: 10: 27, V/V) as an eluting solvent, shaking to elute the template molecules until the template molecules cannot be detected by the high-performance liquid phase of the eluent, washing the eluent to be neutral by using methanol, and drying to obtain the MIPs. The yield thereof was found to be 69.3%.

Example 11 an inventive methodGinsenoside Rg₁Preparation method of molecularly imprinted polymeric material

(1) Mixing 45.75 mg (0.05 mmol) of template molecule ginsenoside Rg₁75.53 mg (0.5 mmol) of functional monomer 4-VPBA is added into 10 mL of absolute ethyl alcohol and dissolved by ultrasonic; adding 132 mu L (0.7 mmol) of crosslinking agent trimethylolpropane trimethacrylate, and stirring at high speed to form a suspension; after adding 0.8 mL of azobisisoheptonitrile (15 mg/mL) as an initiator, nitrogen was purged for at least 30 min to completely remove dissolved oxygen and sealed;

(2) under the protection of nitrogen, the molecularly imprinted polymer is initiated by heat and is placed in a magnetic stirrer with a heating plate for polymerization: and (3) carrying out polymerization reaction for 24 hours at the temperature of 60 ℃ and at the speed of 1500 r/min.

(3) After the polymerization reaction is finished, suction filtration is carried out, and the obtained polymer is dried in a vacuum drying oven for 12 hours at the temperature of 60 ℃. And then, putting the polymer in a constant-temperature culture oscillator by taking methanol-acetic acid-water (63: 10: 27, V/V) as an eluting solvent, shaking to elute the template molecules until the template molecules cannot be detected by the high-performance liquid phase of the eluent, washing the eluent to be neutral by using methanol, and drying to obtain the MIPs. The yield is 73 percent

The above description is only for the preferred embodiment of the present invention, and is not intended to limit the present invention in any way. Those skilled in the art can make many possible variations or modifications to the technical solution of the invention using the methods and techniques described above, or modify equivalent embodiments without departing from the spirit and technical solution of the invention. Therefore, any simple modification, equivalent replacement, and equivalent changes made to the above embodiments without departing from the technical solution of the present invention and the technical essence of the present invention still belong to the protection scope of the present invention.