CN115501649B - Single-column two-phase mixed mode monolithic column and preparation method and application thereof - Google Patents
Single-column two-phase mixed mode monolithic column and preparation method and application thereof Download PDFInfo
- Publication number
- CN115501649B CN115501649B CN202211303261.1A CN202211303261A CN115501649B CN 115501649 B CN115501649 B CN 115501649B CN 202211303261 A CN202211303261 A CN 202211303261A CN 115501649 B CN115501649 B CN 115501649B
- Authority
- CN
- China
- Prior art keywords
- column
- mode
- solution
- dimethylaniline
- phase mixed
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F220/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical or a salt, anhydride ester, amide, imide or nitrile thereof
- C08F220/02—Monocarboxylic acids having less than ten carbon atoms; Derivatives thereof
- C08F220/52—Amides or imides
- C08F220/54—Amides, e.g. N,N-dimethylacrylamide or N-isopropylacrylamide
- C08F220/56—Acrylamide; Methacrylamide
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F222/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by a carboxyl radical and containing at least one other carboxyl radical in the molecule; Salts, anhydrides, esters, amides, imides, or nitriles thereof
- C08F222/10—Esters
- C08F222/1006—Esters of polyhydric alcohols or polyhydric phenols
- C08F222/102—Esters of polyhydric alcohols or polyhydric phenols of dialcohols, e.g. ethylene glycol di(meth)acrylate or 1,4-butanediol dimethacrylate
Landscapes
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Analytical Chemistry (AREA)
- Addition Polymer Or Copolymer, Post-Treatments, Or Chemical Modifications (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention provides a single-column two-phase mixed mode monolithic column, a preparation method and application thereof, wherein the preparation method of the monolithic column comprises the following steps: preparing a hydrophilic mode pre-polymerization liquid and a hydrophobic mode pre-polymerization liquid; and adding N, N-dimethylaniline into the hydrophilic mode prepolymerization liquid and the hydrophobic mode prepolymerization liquid, and then adding the two into the same column tube to obtain the mixed mode monolithic column. The mixed mode monolithic column prepared by the method can simultaneously enrich and purify the licochalcone A and the isoliquiritigenin which are two compounds with different properties in the liquorice extracting solution, so that the enrichment and purification operations of the licochalcone A and the isoliquiritigenin are simpler and more convenient, and the method has the advantages of saving samples, being economical and environment-friendly and the like.
Description
Technical Field
The invention relates to a monolithic column suitable for enrichment and purification of licochalcone A and isoliquiritigenin, in particular to a monolithic column in a single-column two-phase mixed mode, and a preparation method and application thereof.
Background
Monolithic columns, also known as rod columns, continuous beds, and no-plug, are continuous bed stationary phases that polymerize in situ within a chromatographic column using organic or inorganic polymerization methods.
Licochalcone A (Licorice chalcone A, LCA) and isoliquiritigenin are flavonoids extracted from root and stem of common Chinese medicinal Glycyrrhrizae radix, and have multiple pharmacological activities such as anticancer, antiinflammatory, antioxidant and immunity regulating. Among them, their antitumor activity is of greatest concern. However, since licochalcone a and isoliquiritigenin are contained in a medicinal plant in a low amount, extraction, enrichment and purification are difficult. The current separation and purification method of licochalcone A and isoliquiritigenin comprises extraction and column chromatography of macroporous resin, polyamide, silica gel and the like. However, these methods are complicated in operation, low in recovery rate of licochalcone A and isoliquiritigenin, poor in separation effect, and high in organic residues. Therefore, a simple and efficient method for enriching and purifying licochalcone A and isoliquiritigenin is needed to be found.
Disclosure of Invention
The invention aims to provide a preparation method of a single-column two-phase mixed mode monolithic column.
The second objective of the present invention is to provide a monolithic column with a single-column two-phase mixed mode.
The invention aims at providing a method for enriching and purifying licochalcone A and isoliquiritigenin in liquorice.
One of the objects of the present invention is achieved by: the preparation method of the single-column two-phase mixed mode monolithic column comprises the following steps:
a. mixing benzoyl peroxide, acrylamide, ethylene glycol dimethacrylate and n-propanol to obtain a hydrophilic mode prepolymer; to be functionalized MIL-125-NH 2 Mixing benzoyl peroxide, isooctyl methacrylate, ethylene glycol dimethacrylate, n-hexanol and n-butanol to obtain a hydrophobic mode prepolymer;
b. carrying out ultrasonic treatment on the hydrophilic mode prepolymer liquid, and adding N, N-dimethylaniline after ultrasonic treatment to obtain a solution A; carrying out ultrasonic treatment on the pre-polymerization solution in a hydrophobic mode, and adding N, N-dimethylaniline after ultrasonic treatment to obtain a solution B; firstly, filling the solution A into a chromatographic column tube, then pouring the solution B into the same chromatographic column tube, sealing the chromatographic column tube, carrying out water bath at 25-45 ℃ for 3-4 hours, taking out and flushing to obtain the single-column two-phase mixed mode monolithic column.
The dosage ratio of benzoyl peroxide, acrylamide, glycol dimethacrylate, N-propanol and N, N-dimethylaniline in the hydrophilic mode prepolymer liquid in the step a is 0.001-0.1g:0.01-0.02g:0.4-0.5mL:0.8-0.9mL:10-50 mu L;
functionalized MIL-125-NH in the hydrophobic mode pre-polymerization solution 2 Benzoyl peroxide, isooctyl methacrylate, ethylene glycol dimethacrylate, N-hexanol, N-butanol and N, N-dimethylaniline in an amount ratio of 0.001-0.01g:0.001-0.1g:0.1-0.3mL:0.4-0.5mL:0.8-0.9mL:0.4-0.5mL:10-50 mu L.
The dosage ratio of benzoyl peroxide, acrylamide, ethylene glycol dimethacrylate, N-propanol and N, N-dimethylaniline in the hydrophilic mode pre-polymerization solution in the step a is 0.003g:0.018g:0.45mL:0.85mL: 30. Mu.L;
functionalized MIL-125-NH in the hydrophobic mode pre-polymerization solution 2 Benzoyl peroxide, isooctyl methacrylate, ethylene glycol dimethacrylate, N-hexanol, N-butanol and N, N-dimethylaniline in an amount ratio of 0.003g:0.003g:0.2mL:0.45mL:0.8mL:0.4mL: 30. Mu.L;
in the step B, the volume ratio of the solution A to the solution B is 1/2-2/1.
The second object of the invention is realized in such a way that: a single-column two-phase mixed mode monolithic column prepared by any one of the methods.
The third object of the present invention is achieved by: an enrichment and purification method of licochalcone A and isoliquiritigenin in Glycyrrhrizae radix comprises combining single column two-phase mixed mode monolithic column with high performance liquid chromatography, and purifying and enriching licochalcone A and isoliquiritigenin on line.
A mixed solution of acetonitrile and water is adopted as a mobile phase, and the volume ratio of the acetonitrile to the water is 30:70.
The single-column two-phase mixed mode monolithic column prepared by the method has more excellent enrichment and purification capacity than the single-mode monolithic column, and has good permeability and lower mass transfer resistance, and the preparation method is simple, economical and practical. The single-column two-phase mixed mode monolithic column can realize the enrichment and purification of licochalcone A and isoliquiritigenin in liquorice at the same time, and an interface is not needed in the enrichment and purification process, so that the monolithic column has the advantages of simplicity and convenience in operation, sample saving, economy, environmental friendliness and the like.
Drawings
FIG. 1 is an electron microscopic view of a hydrophilic part (A) and a hydrophobic part (B) of a single-column two-phase mixed mode monolithic column prepared in example 1.
FIG. 2 is a chromatogram of a single column two-phase mixed mode monolith prepared with different ratios between hydrophilic and hydrophobic portions of the monolith and enriched and purified licorice extract from other chromatographic columns. Wherein a, b and c are separation results after enrichment and purification of the single-column two-phase liquid chromatography monolithic columns, and the volume ratio of the hydrophilic material to the hydrophobic material of the three single-column two-phase liquid chromatography monolithic columns is 2:1, 1:2 and 1:1 respectively; d, enriching and purifying the separation result by a hydrophilic mode monolithic column; e, enriching and purifying the separation result by a hydrophobic mode monolithic column.
FIG. 3 is a separation comparison chromatogram of the licorice extract of example 4 under the same conditions, through a single column two-phase mixed mode monolith (a) and an untreated licorice extract (b).
FIG. 4 is a hydrophobic mode monolithic column electron microscopy image of example 10.
Detailed Description
The invention is further illustrated by the following examples, which are given by way of illustration only and are not intended to limit the scope of the invention in any way.
The procedures and methods not described in detail in the examples below are conventional methods well known in the art, and the reagents used in the examples are all analytically or chemically pure and are either commercially available or prepared by methods well known to those of ordinary skill in the art.
Example 1 preparation method of Single column two-phase Mixed mode monolithic column (method 1)
S1, precisely weighing 0.003g of benzoyl peroxide, 0.018g of acrylamide, 0.45mL of ethylene glycol dimethacrylate and 0.85mL of n-propanol, sequentially adding into a clean No. 1 centrifuge tube, and carrying out ultrasonic vibration to obtain a prepolymer liquid in a hydrophilic mode;
precisely weighing 0.003g of functionalized MIL-125-NH 2 0.003g of benzoyl peroxide, 0.2mL of isooctyl methacrylate, 0.45mL of ethylene glycol dimethacrylate, 0.8mL of n-hexanol and 0.4mL of n-butanol are sequentially added into a clean centrifuge tube No. 2, shaken well and subjected to ultrasonic oscillation.
S2, after the solid of the hydrophilic mode prepolymer in the centrifuge tube No. 1 is dissolved and uniformly dispersed, adding 30 mu L of N, N-dimethylaniline, rapidly and uniformly mixing, and carrying out ultrasonic degassing to obtain a solution A; after the solid of the pre-polymerization liquid in the hydrophobic mode in the centrifuge tube No. 2 is dissolved and uniformly dispersed, 30 mu L of N, N-dimethylaniline is added, the mixture is rapidly and uniformly mixed, and the ultrasonic degassing is carried out, so that a solution B is obtained. Then the solution A and the solutionPouring the liquid B into a hollow column tube with one end sealed, sealing the other end, and 30 o And C, carrying out water bath reaction for 3.5h to obtain the monolithic column with the single-column two-phase mixed mode. In this example, the volume ratio of solution A to solution B was 2:1.
Example 2 preparation of Single column two-phase Mixed mode monolithic column (method 2)
This example is identical to the preparation of example 1, except that the volume ratio of solution A to solution B is 1:2.
Example 3 preparation of Single column two-phase Mixed mode monolithic column (method 3)
This example is identical to the preparation of example 1, except that the volume ratio of solution A to solution B is 1:1.
The single-column two-phase mixed mode monolithic column prepared by the three methods is respectively connected with a high-efficiency liquid phase system, acetonitrile and water are used as mobile phases, under the condition of acetonitrile/water (30/70, V/V), licorice extracting solution is injected, acetonitrile/water (90/10, V/V) is used for eluting, eluent is collected, 40uL of methanol is added after drying, and solutions a, b and c are prepared. In addition, a hydrophilic mode monolith was prepared using the hydrophilic mode pre-polymerization solution of example 1, a hydrophobic mode monolith was prepared using the hydrophobic mode pre-polymerization solution of example 1, the hydrophilic mode monolith and the hydrophobic mode monolith were used as a control, and were connected to a high performance liquid phase system under the same conditions, acetonitrile and water were used as mobile phases, licorice extract was injected, acetonitrile/water (90/10, V/V) was used for elution under acetonitrile/water (30/70, V/V) conditions, and 40uL of methanol was added after drying to prepare solutions d and e. The solution a, b, C, d, e was analyzed with a C18 analytical column.
The analysis results are shown in FIG. 4. From the figure, it can be seen that the mixed mode monolithic column with the hydrophilic part and the hydrophobic part being 1:1 has the highest enrichment capacity for isoliquiritigenin and good impurity removal effect. The microscopic morphology of the hydrophilic part of the single-column two-phase mixed mode monolithic column prepared by the scanning electron microscope observation method 1 is shown in fig. 1 (a), and the microscopic morphology of the hydrophobic part of the single-column two-phase mixed mode monolithic column prepared by the scanning electron microscope observation method 1 is shown in fig. 1 (B).
Example 4 Single column two phase Mixed mode monolithic column for enrichment and purification of licochalcone A and isoliquiritigenin
The single-column two-phase mixed mode monolithic column prepared by the method 1 is connected to a high performance liquid chromatograph, acetonitrile and water (30:70, v/v) are used as mobile phases, licochalcone A and isoliquiritigenin in a liquorice extract sample are enriched, acetonitrile and water (90:10, v/v) are used as elution flows to relatively enrich licochalcone A and isoliquiritigenin on the monolithic column, eluent containing target substances is collected, the eluent is concentrated to obtain licochalcone A and isoliquiritigenin solution, liquorice extract which is not treated by the monolithic column is used as a reference, and C18 analytical columns are used for analysis under the same conditions, and the obtained comparison results are shown in figure 3.
Example 5 preparation of Single column two-phase Mixed mode monolithic column (method 4)
S1, precisely weighing 0.003g of benzoyl peroxide, 0.018g of acrylamide, 0.45mL of ethylene glycol dimethacrylate and 0.8mL of n-propanol, sequentially adding into a clean No. 1 centrifuge tube, and carrying out ultrasonic vibration to obtain a prepolymer liquid in a hydrophilic mode;
precisely weighing 0.003g of functionalized MIL-125-NH 2 0.003g of benzoyl peroxide, 0.2mL of isooctyl methacrylate, 0.45mL of ethylene glycol dimethacrylate, 0.8mL of n-hexanol and 0.4mL of n-butanol are sequentially added into a clean centrifuge tube No. 2, shaken well and subjected to ultrasonic oscillation.
S2, after the solid of the hydrophilic mode prepolymer in the centrifuge tube No. 1 is dissolved and uniformly dispersed, adding 30 mu L of N, N-dimethylaniline, rapidly and uniformly mixing, and carrying out ultrasonic degassing to obtain a solution A; after the solid of the pre-polymerization liquid in the hydrophobic mode in the centrifuge tube No. 2 is dissolved and uniformly dispersed, 30 mu L of N, N-dimethylaniline is added, the mixture is rapidly and uniformly mixed, and the ultrasonic degassing is carried out, so that a solution B is obtained. Then pouring the solution A and the solution B into a hollow column tube with one end sealed, sealing the other end, 30 o The C water bath reaction is carried out for 3.5 hours to obtain single-column two-phase mixtureA modular monolithic column. In this example, the volume ratio of solution A to solution B was 1:1.
Example 6 preparation of Single column two-phase Mixed mode monolithic column (method 5)
S1, precisely weighing 0.003g of benzoyl peroxide, 0.013g of acrylamide, 0.45mL of ethylene glycol dimethacrylate and 0.85mL of n-propanol, sequentially adding into a clean No. 1 centrifuge tube, and carrying out ultrasonic vibration to obtain a prepolymer liquid in a hydrophilic mode;
precisely weighing 0.003g of functionalized MIL-125-NH 2 0.003g of benzoyl peroxide, 0.2mL of isooctyl methacrylate, 0.45mL of ethylene glycol dimethacrylate, 0.8mL of n-hexanol and 0.4mL of n-butanol are sequentially added into a clean centrifuge tube No. 2, shaken well and subjected to ultrasonic oscillation.
S2, after the solid of the hydrophilic mode prepolymer in the centrifuge tube No. 1 is dissolved and uniformly dispersed, adding 30 mu L of N, N-dimethylaniline, rapidly and uniformly mixing, and carrying out ultrasonic degassing to obtain a solution A; after the solid of the pre-polymerization liquid in the hydrophobic mode in the centrifuge tube No. 2 is dissolved and uniformly dispersed, 30 mu L of N, N-dimethylaniline is added, the mixture is rapidly and uniformly mixed, and the ultrasonic degassing is carried out, so that a solution B is obtained. Then pouring the solution A and the solution B into a hollow column tube with one end sealed, sealing the other end, 30 o And C, carrying out water bath reaction for 3.5h to obtain the monolithic column with the single-column two-phase mixed mode. In this example, the volume ratio of solution A to solution B was 1:1.
Example 7 preparation of Single column two-phase Mixed mode monolithic column (method 6)
S1, precisely weighing 0.003g of benzoyl peroxide, 0.018g of acrylamide, 0.5mL of ethylene glycol dimethacrylate and 0.85mL of n-propanol, sequentially adding into a clean No. 1 centrifuge tube, and carrying out ultrasonic vibration to obtain a prepolymer liquid in a hydrophilic mode;
precisely weighing 0.003g of functionalized MIL-125-NH 2 0.003g of benzoyl peroxide, 0.2mL of isooctyl methacrylate, 0.45mL of ethylene glycol dimethacrylate, 0.8mL of n-hexanol and 0.5mL of n-butanol are sequentially added into a clean centrifuge tube No. 2, shaken well and subjected to ultrasonic oscillation.
S2, adding 30 mu L of solid solution after the solid solution of the hydrophilic mode prepolymer in the centrifuge tube No. 1 is dissolved and uniformly dispersedN, N-dimethylaniline is quickly and evenly mixed, and is subjected to ultrasonic degassing to obtain a solution A; after the solid of the pre-polymerization liquid in the hydrophobic mode in the centrifuge tube No. 2 is dissolved and uniformly dispersed, 30 mu L of N, N-dimethylaniline is added, the mixture is rapidly and uniformly mixed, and the ultrasonic degassing is carried out, so that a solution B is obtained. Then pouring the solution A and the solution B into a hollow column tube with one end sealed, sealing the other end, 30 o And C, carrying out water bath reaction for 3.5h to obtain the monolithic column with the single-column two-phase mixed mode. In this example, the volume ratio of solution A to solution B was 1:1.
The three above-mentioned monolithic columns were used for enrichment and purification of licochalcone a and isoliquiritigenin, and were respectively analyzed under the same conditions using a C18 analytical column as a control with the licorice extract not subjected to monolithic column treatment, and the comparative results were the same as those obtained in example 4.
Example 8 preparation of hydrophilic mode monolith (a)
0.003g of benzoyl peroxide, 0.018g of acrylamide, 0.45mL of ethylene glycol dimethacrylate and 0.85mL of n-propanol are sequentially weighed into a clean centrifuge tube, and the initiator is completely dissolved by ultrasonic vibration. Then 30. Mu.L of N, N-dimethylaniline was added, mixed by shaking and degassed by ultrasound. Then pouring the mixed solution into a hollow column tube with one end sealed, sealing the other end, 30 o And C, carrying out water bath reaction for 3.5h to obtain the hydrophilic mode monolithic column a.
EXAMPLE 9 preparation of hydrophilic mode monolith (b)
Under the same conditions as in example 8, 0.003g of benzoyl peroxide, 0.013g of acrylamide, 0.45mL of ethylene glycol dimethacrylate and 0.85mL of n-propanol are sequentially weighed into a clean centrifuge tube, and the initiator is completely dissolved by ultrasonic vibration. Then 30. Mu.L of N, N-dimethylaniline was added, mixed by shaking and degassed by ultrasound. Then pouring the mixed solution into a hollow column tube with one end sealed, sealing the other end, 30 o And C, carrying out water bath reaction for 3.5h to obtain the hydrophilic mode monolithic column b.
EXAMPLE 10 preparation of hydrophilic mode monolith (c)
Under the same conditions as in example 8, 0.003g of benzoyl peroxide, 0.018g of acrylamide and 0.45mL of ethyl acetate were precisely weighedGlycol dimethacrylate, 1.5mL of n-propanol was added to a clean centrifuge tube in sequence, and the initiator was completely dissolved by ultrasonic vibration. Then 30. Mu.L of N, N-dimethylaniline was added, mixed by shaking and degassed by ultrasound. Then pouring the mixed solution into a hollow column tube with one end sealed, sealing the other end, 30 o And (3) carrying out water bath reaction on the C for 3.5 hours to obtain the hydrophilic mode monolithic column C.
EXAMPLE 11 preparation of hydrophilic mode monolith (d)
Under the same conditions as in example 8, 0.003g of benzoyl peroxide, 0.018g of acrylamide, 0.5mL of ethylene glycol dimethacrylate and 0.85mL of n-propanol were weighed precisely, sequentially added into a clean centrifuge tube, and subjected to ultrasonic vibration to completely dissolve the initiator. Then 30. Mu.L of N, N-dimethylaniline was added, mixed by shaking and degassed by ultrasound. Then pouring the mixed solution into a hollow column tube with one end sealed, sealing the other end, 30 o And (3) carrying out water bath reaction on the C for 3.5 hours to obtain the hydrophilic mode monolithic column d.
The four hydrophilic mode monolithic columns are connected with a high performance liquid phase system, 100% methanol is used as a mobile phase to be washed for 1.5 hours, and unreacted soluble components such as pore-forming agents are removed. The back pressure of the whole column a was measured to be 2.7MPa, and the theoretical plate number was measured to be 15200 plates/m. The specific surface area of the monolithic column a was 8.5543m by a specific surface area meter 2 And/g. The column back pressure of the whole column b was 5.3MPa, the theoretical plate number was 2880 plates/m, and the specific surface area was 5.2544 m 2 And/g. The column back pressure of the whole column b increases and the theoretical plate number decreases due to the decrease in the amount of monomer. Higher column back pressures may collapse the monolith. The whole column c is flushed out in the flushing process, and subsequent experiments cannot be performed. Indicating an increase in porogen and a decrease in the mechanical strength of the monolith. The column back pressure of the whole column d was measured to be 5.7MPa, and the theoretical plate number was measured to be 1340 plates/m. The increase in the crosslinking agent increases the column back pressure of the monolithic column d, which increases the system pressure and may adversely affect the liquid phase system. Therefore, the hydrophilic mode monolith a has the best hydrophilic effect and is the optimal hydrophilic mode monolith.
EXAMPLE 12 preparation of hydrophobic mode monolith (e)
Precisely weigh 0.003g functionChemical MIL-125-NH 2 0.003g of benzoyl peroxide, 0.2mL of isooctyl methacrylate, 0.45mL of ethylene glycol dimethacrylate, 0.8mL of n-hexanol and 0.4mL of n-butanol are sequentially added into a clean centrifuge tube, shaken well, and ultrasonically oscillated until the solids in the tube are dissolved and dispersed well. Then 30. Mu.L of N, N-dimethylaniline was added, mixed by shaking and degassed by ultrasound. Then pouring the mixed solution into a hollow column tube with one end sealed, sealing the other end, 30 o And C, carrying out water bath reaction for 3.5h to obtain the hydrophobic mode monolithic column e.
EXAMPLE 13 preparation of hydrophobic mode monolith (f)
Under the same conditions as in example 12, 0.003g of benzoyl peroxide, 0.2mL of isooctyl methacrylate, 0.45mL of ethylene glycol dimethacrylate, 0.8mL of n-hexanol and 0.4mL of n-butanol were weighed precisely, sequentially added into a clean centrifuge tube, shaken well, and vibrated ultrasonically until the solids in the tube were dissolved and dispersed well. Then 30. Mu.L of N, N-dimethylaniline was added, mixed by shaking and degassed by ultrasound. Then pouring the mixed solution into a hollow column tube with one end sealed, sealing the other end, 30 o And C, carrying out water bath reaction for 3.5h to obtain the hydrophobic mode monolithic column f.
EXAMPLE 14 preparation of hydrophobic mode monolith (g)
Under the same conditions as in example 12, 0.003g of functionalized MIL-125-NH was precisely weighed 2 0.003g of benzoyl peroxide, 0.2mL of isooctyl methacrylate, 0.45mL of ethylene glycol dimethacrylate, 0.8mL of n-hexanol and 0.8mL of n-butanol are sequentially added into a clean centrifuge tube, shaken well, and ultrasonically oscillated until the solids in the tube are dissolved and dispersed well. Then 30. Mu.L of N, N-dimethylaniline was added, mixed by shaking and degassed by ultrasound. Then pouring the mixed solution into a hollow column tube with one end sealed, sealing the other end, 30 o And C, carrying out water bath reaction for 3.5h to obtain the hydrophobic mode monolithic column g.
EXAMPLE 15 preparation of hydrophobic mode monolith (h)
Under the same conditions as in example 12, 0.003g of functionalized MIL-125-NH 2 0.003g benzoyl peroxide, 0.2mL isooctyl methacrylate, 0.5mL ethylene glycol dimethacrylate, 0.8mL n-butyl acrylateHexanol and 0.4mL of n-butanol were added sequentially to a clean centrifuge tube, shaken well, and sonicated until the solids in the tube were dissolved and dispersed well. Then 30. Mu.L of N, N-dimethylaniline was added, mixed by shaking and degassed by ultrasound. Then pouring the mixed solution into a hollow column tube with one end sealed, sealing the other end, 30 o And C, carrying out water bath reaction for 3.5h to obtain the hydrophobic mode monolithic column h.
The four hydrophobic mode monolithic columns are connected with a high performance liquid phase system, and are washed for 1.5 hours by taking 100% methanol as a mobile phase, so that unreacted soluble components such as a pore-forming agent and the like are removed. The back pressure of the whole column e was measured to be 1.9MPa, and the theoretical plate number was measured to be 17800 plates/m. The specific surface area of the monolithic column e was 29.9273m by a specific surface area meter 2 And/g. The column back pressure of the whole column f was 3.6MPa, the theoretical plate number was 5440 plates/m, and the specific surface area was 10.1181 m 2 And/g. The microscopic morphology was observed with a scanning electron microscope as shown in fig. 4. As can be seen by comparing FIG. 1 (B), MIL-125-NH was added 2 The pore size distribution of the hydrophobic mode monolithic column is more uniform. Since MIL-125-NH is not added 2 The specific surface area of the monolithic column f is reduced, which is not beneficial to enriching and purifying licochalcone A and isoliquiritigenin. The whole column g is flushed out in the flushing process, and subsequent experiments cannot be performed. Indicating an increase in porogen and a decrease in the mechanical strength of the monolith. The column back pressure of the whole column h was measured to be 4.1MPa, and the theoretical plate number was measured to be 3960 plates/m. The increase of the cross-linking agent causes the increase of the column back pressure of the whole column, and the column back pressure is increased according to Darcy's law, so that the permeability is correspondingly reduced, and the enrichment and purification of licochalcone A and isoliquiritigenin in liquorice are not facilitated. Therefore, the hydrophobic mode monolith e has the best hydrophobic effect and is the optimal hydrophobic mode monolith.
Claims (6)
1. The preparation method of the single-column two-phase mixed mode monolithic column is characterized by comprising the following steps of:
a. mixing benzoyl peroxide, acrylamide, ethylene glycol dimethacrylate and n-propanol to obtain a hydrophilic mode prepolymer; to be functionalized MIL-125-NH 2 Benzoyl peroxide, isooctyl methacrylateMixing ester, ethylene glycol dimethacrylate, n-hexanol and n-butanol to obtain a hydrophobic mode prepolymer;
b. carrying out ultrasonic treatment on the hydrophilic mode prepolymer liquid, and adding N, N-dimethylaniline after ultrasonic treatment to obtain a solution A; carrying out ultrasonic treatment on the pre-polymerization solution in a hydrophobic mode, and adding N, N-dimethylaniline after ultrasonic treatment to obtain a solution B; firstly, filling a solution A into a chromatographic column tube, then pouring a solution B into the same chromatographic column tube, sealing the chromatographic column tube, carrying out water bath at 25-45 ℃ for 3-4 hours, and taking out and flushing to obtain a single-column two-phase mixed mode monolithic column;
the dosage ratio of benzoyl peroxide, acrylamide, glycol dimethacrylate, N-propanol and N, N-dimethylaniline in the hydrophilic mode prepolymer liquid in the step a is 0.001-0.1g:0.018g:0.45mL:0.8-0.9mL:10-50 mu L;
functionalized MIL-125-NH in the hydrophobic mode pre-polymerization solution 2 Benzoyl peroxide, isooctyl methacrylate, ethylene glycol dimethacrylate, N-hexanol, N-butanol and N, N-dimethylaniline in an amount ratio of 0.001-0.01g:0.001-0.1g:0.1-0.3mL:0.45mL:0.8-0.9mL:0.4-0.5mL:10-50 mu L;
the prepared single-column two-phase mixed mode monolithic column is used for purifying and enriching licochalcone A and isoliquiritigenin in liquorice on line.
2. The method for preparing a single-column two-phase mixed mode monolithic column according to claim 1, wherein the dosage ratio of benzoyl peroxide, acrylamide, ethylene glycol dimethacrylate, N-propanol and N, N-dimethylaniline in the hydrophilic mode pre-polymerization solution in the step a is 0.003g:0.018g:0.45mL:0.85mL: 30. Mu.L;
functionalized MIL-125-NH in the hydrophobic mode pre-polymerization solution 2 Benzoyl peroxide, isooctyl methacrylate, ethylene glycol dimethacrylate, N-hexanol, N-butanol and N, N-dimethylaniline in an amount ratio of 0.003g:0.003g:0.2mL:0.45mL:0.8mL:0.4mL: 30. Mu.L.
3. The method for preparing a monolithic column in a single-column two-phase mixed mode according to claim 1, wherein the volume ratio of the solution A to the solution B in the step B is 1/2 to 2/1.
4. A single-column two-phase mixed mode monolithic column, characterized in that it is prepared by the method of any one of claims 1-3.
5. An enrichment and purification method of licochalcone A and isoliquiritigenin in liquorice is characterized in that single-column two-phase mixed mode monolithic column in claim 4 is combined with high performance liquid chromatography to carry out online purification and enrichment of licochalcone A and isoliquiritigenin.
6. The method for enriching and purifying licochalcone A and isoliquiritigenin from licorice according to claim 5, wherein a mixed solution of acetonitrile and water is used as a mobile phase, and the volume ratio of acetonitrile to water is 30:70.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211303261.1A CN115501649B (en) | 2022-10-24 | 2022-10-24 | Single-column two-phase mixed mode monolithic column and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211303261.1A CN115501649B (en) | 2022-10-24 | 2022-10-24 | Single-column two-phase mixed mode monolithic column and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115501649A CN115501649A (en) | 2022-12-23 |
| CN115501649B true CN115501649B (en) | 2023-06-20 |
Family
ID=84512028
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211303261.1A Active CN115501649B (en) | 2022-10-24 | 2022-10-24 | Single-column two-phase mixed mode monolithic column and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115501649B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110724228A (en) * | 2019-11-19 | 2020-01-24 | 常州大学 | Preparation method of shape memory imprinting gel |
| CN111871394A (en) * | 2020-07-30 | 2020-11-03 | 河南省农产品质量安全检测中心(河南省绿色食品发展中心) | Novel bifunctional group mixed mode organic polymer monolithic column, preparation method and application thereof |
| CN113234192A (en) * | 2021-05-19 | 2021-08-10 | 福州大学 | Double-template surface molecularly imprinted nano composite material, preparation method and application thereof |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2419886B (en) * | 2003-05-28 | 2008-12-17 | Waters Investments Ltd | Novel nanocomposites and their application as monolith columns |
| US10119944B2 (en) * | 2013-12-24 | 2018-11-06 | Waters Technologies Corporation | Materials for hydrophilic interaction chromatography and processes for preparation and use thereof for analysis of glycoproteins and glycopeptides |
-
2022
- 2022-10-24 CN CN202211303261.1A patent/CN115501649B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110724228A (en) * | 2019-11-19 | 2020-01-24 | 常州大学 | Preparation method of shape memory imprinting gel |
| CN111871394A (en) * | 2020-07-30 | 2020-11-03 | 河南省农产品质量安全检测中心(河南省绿色食品发展中心) | Novel bifunctional group mixed mode organic polymer monolithic column, preparation method and application thereof |
| CN113234192A (en) * | 2021-05-19 | 2021-08-10 | 福州大学 | Double-template surface molecularly imprinted nano composite material, preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115501649A (en) | 2022-12-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102068968B (en) | Lead ion imprinting integral column and preparation method thereof | |
| CN111470953A (en) | Method for extracting and separating high-purity cannabidiol from low-content industrial cannabis sativa leaves | |
| EP0609373A1 (en) | Column with microporous polymer media. | |
| CN112608404B (en) | Amino-functionalized heterostructure porous microsphere and preparation method and application thereof | |
| CN1680392A (en) | Preparation of high-purity ginkgolide | |
| Schulte et al. | Preparative monolithic silica sorbents (PrepRODs™) and their use in preparative liquid chromatography | |
| CN115501649B (en) | Single-column two-phase mixed mode monolithic column and preparation method and application thereof | |
| CN111871394A (en) | Novel bifunctional group mixed mode organic polymer monolithic column, preparation method and application thereof | |
| CN101230080A (en) | simulated moving bed chromatography separation of 20(S) and 20(R)-ginsenoside Rg3 enantiomer | |
| Takeuchi et al. | Miniaturized molecularly imprinted continuous polymer rods | |
| Ciccioli et al. | Graphitized carbon black columns for high-performance liquid chromatography | |
| CN110922439A (en) | Method for separating and preparing gram-grade high-purity natural product | |
| CN115487543B (en) | Single-column two-phase liquid chromatography monolithic column and preparation method and application thereof | |
| CN109651301A (en) | A kind of purification process of orlistat | |
| Svec et al. | Molded separation media: An inexpensive, efficient, and versatile alternative to packed columns for the fast HPLC separation of peptides, proteins, and synthetic oligomers and polymers | |
| CN104557542B (en) | A kind of Supercritical fluid chromatography prepares the method for high purity EPA ester and DHA ester monomer | |
| CN115260001B (en) | Preparation method of (1S, 2S, 4R) -dipentene-1, 2-diol | |
| CN107376420A (en) | Disperse mobile phase countercurrent chromatography separation method | |
| CN102336792A (en) | Three-zone simulated moving bed chromatography method for separating and purifying paeoniflorin | |
| CN111790360A (en) | Pore-foaming agent, zinc ion imprinted polymer and preparation method thereof, zinc ion imprinted monolithic column and preparation method thereof | |
| Shi et al. | Fabrication and characterization of a novel biporous spherical adsorbent for protein chromatography | |
| CN101625347A (en) | Method for synthesizing integral bed for plasmid analysis | |
| CN114405063B (en) | Method for enriching and purifying diosmetin from dandelion on line | |
| Chen et al. | Novel liquid-liquid-solid microextraction using molecularly imprinted polymer monolithic fibres and its application to the extraction of s-triazine herbicides from water samples | |
| CN111686136B (en) | Method for purifying triterpenoids |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |