CN112604672B - Functionalized carboxymethyl chitosan composite adsorbent and preparation method and application thereof - Google Patents
Functionalized carboxymethyl chitosan composite adsorbent and preparation method and application thereof Download PDFInfo
- Publication number
- CN112604672B CN112604672B CN202011383247.8A CN202011383247A CN112604672B CN 112604672 B CN112604672 B CN 112604672B CN 202011383247 A CN202011383247 A CN 202011383247A CN 112604672 B CN112604672 B CN 112604672B
- Authority
- CN
- China
- Prior art keywords
- carboxymethyl chitosan
- adsorption
- composite adsorbent
- functionalized
- chitosan composite
- 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
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/26—Synthetic macromolecular compounds
- B01J20/265—Synthetic macromolecular compounds modified or post-treated polymers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/30—Processes for preparing, regenerating, or reactivating
- B01J20/3085—Chemical treatments not covered by groups B01J20/3007 - B01J20/3078
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H15/00—Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
- C07H15/18—Acyclic radicals, substituted by carbocyclic rings
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
The invention provides a preparation method of a functionalized carboxymethyl chitosan composite adsorbent, which comprises the steps of taking carboxymethyl chitosan and polyethyleneimine as raw materials, adding a dispersing agent and a cross-linking agent, carrying out Schiff base reaction to obtain pink flocculent precipitate, adding a reducing agent into the pink flocculent precipitate, and reducing imino groups into amino groups to obtain the functionalized carboxymethyl chitosan composite adsorbent. The invention also provides application of the adsorbent in efficiently separating phenylethanoid glycosides compounds in cistanche tubulosa. The invention applies an adsorption method to the separation of the active ingredients of the natural product, designs the functionalized composite adsorbent for efficiently separating the phenylethanoid glycosides compounds in the cistanche tubulosa, and provides a new method for efficiently separating the phenylethanoid glycosides in the active ingredients of the natural product.
Description
Technical Field
The invention relates to a functionalized carboxymethyl chitosan composite adsorbent and a preparation method and application thereof.
Background
Cistanche tubulosa is a perennial parasitic plant that grows by absorbing nutrients from host plants, mainly including sand-grown guayule, chinese-grown guayule, and white-flower guayule. Meanwhile, cistanche tubulosa is a precious nourishing Chinese herbal medicine, is known as 'desert ginseng', and has the functions of nourishing kidney, resisting aging, enhancing blood essence, moistening large intestine and smoothing stool. Studies show that phenylethanoid glycosides are the main active ingredients of cistanche tubulosa, have many hydroxyl groups and phenolic hydroxyl groups on molecules, exist in various plants, and have the effects of neuroprotection, immunoregulation, anti-inflammation, liver protection, anti-tumor and antioxidation. The content of echinacoside and verbascoside is the highest in the phenylethanoid glycosides of cistanche tubulosa, and the echinacoside and the verbascoside are index components for quality identification of cistanche tubulosa. The natural product system is a complex system, which comprises thousands of molecules, and the contents of the molecules are very different, and a plurality of components have similar structures, so that the separation difficulty is high. The purity requirement of active substances of natural products makes the selective separation of single substances more and more important, and the existing separation and purification method of active ingredients of natural products mainly comprises the following steps: adsorption, molecular imprinting, solvent extraction, membrane separation, and the like. The adsorption method is one of the ideal methods for separating the effective components of natural products, and comprises the steps of adsorbing one or more components in a water sample on the surface by using a porous solid adsorbent, desorbing the predicted components by using a suitable solvent, heating or blowing and the like to achieve the purpose of separating and enriching target components.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides the functionalized carboxymethyl chitosan composite adsorbent which is environment-friendly, low in energy consumption, simple in preparation method and high in separation efficiency, and the functionalized carboxymethyl chitosan composite adsorbent is used for separating the phenylethanoid glycosides compounds in cistanche tubulosa.
The invention provides a preparation method of a functionalized carboxymethyl chitosan composite adsorbent, which comprises the steps of taking carboxymethyl chitosan and polyethyleneimine as raw materials, adding a dispersant and a cross-linking agent, carrying out Schiff base reaction to obtain pink flocculent precipitate, preferably, adding a reducing agent into the pink flocculent precipitate after the pink flocculent precipitate is obtained, and reducing imino groups into amino groups to obtain the functionalized carboxymethyl chitosan composite adsorbent.
Preferably, the weight ratio of the carboxymethyl chitosan to the polyethyleneimine is 1: 1.5-6; preferably 1: 4.5-6; most preferably 1: 4.5.
And changing the addition of the polyethyleneimine, keeping the addition of other monomers unchanged, and obtaining the functionalized carboxymethyl chitosan composite adsorbent with different polyethyleneimine grafting amounts under the same reaction conditions.
Preferably, the dispersant is polyvinyl alcohol; and/or
The cross-linking agent is glutaraldehyde; and/or
The reducing agent is sodium cyanoborohydride; and/or
The schiff base reaction is completed at room temperature.
Preferably, the degree of substitution of the carboxymethyl chitosan is not less than 80%.
Preferably, the molecular weight of the polyethyleneimine is 1800-70000.
Preferably, the hydrolysis degree of the polyvinyl alcohol is 99%, and the average polymerization degree is 1700.
The invention also provides a functionalized carboxymethyl chitosan composite adsorbent prepared by any one of the methods.
The invention also provides the application of the functionalized carboxymethyl chitosan composite adsorbent in the efficient separation of the phenylethanoid glycosides compounds in cistanche tubulosa; the phenylethanoid glycosides compounds are echinacoside and verbascoside.
Preferably, the application specifically comprises: adding a functionalized carboxymethyl chitosan composite adsorbent into a phenylethanoid glycoside solution, and fully oscillating for adsorption; preferably, the adsorption conditions are: 150 ℃ and 200rpm, and the adsorption is carried out for 12-24h under 30 ℃ oscillation.
Preferably, the functionalized carboxymethyl chitosan composite adsorbent is eluted after adsorption, and then the phenylethanol glycoside compounds are adsorbed again; the elution conditions were: the eluent is: methanol: acetic acid: the volume ratio of water is 1:1: 8; elution was carried out at 45 ℃ for 6-8h with shaking at 150-.
The adsorption method is applied to the separation of the active ingredients of the natural product, the functionalized composite adsorbent is designed to be used for efficiently separating the phenylethanoid glycosides compounds in the cistanche tubulosa, and the phenylethanoid glycosides compounds in the cistanche tubulosa can be separated again after being desorbed, so that a new method is provided for efficiently separating the phenylethanoid glycosides in the active ingredients of the natural product.
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples are commercially available unless otherwise specified.
The preparation method of the functionalized carboxymethyl chitosan composite adsorbent comprises the following steps:
1g of carboxymethyl chitosan, 1.5-6.0g of 50 wt% polyethyleneimine aqueous solution and 0.4-0.6g of polyvinyl alcohol are dissolved in 130mL of 110-one-water ultrapure water, 15-20mL of 5 wt% glutaraldehyde aqueous solution is dropwise added into the solution (dropwise addition is completed within 30min), and the solution is vigorously stirred for 5-7h at room temperature (Schiff base reaction is completed) to obtain pink flocculent precipitate. After the reaction is vigorously stirred for 5-7h, adding sodium cyanoborohydride into the reaction, and adding 40-60mg every 2-3h for 3 times in total. After the reaction is finished, washing with deionized water for several times, and freezing in vacuum for 20-24h to obtain the functionalized carboxymethyl chitosan composite adsorbent.
Wherein, the substitution degree of the carboxymethyl chitosan is not less than 80% (which is beneficial to forming a uniform network structure).
The molecular weight of the polyethyleneimine is 1800-.
The hydrolysis degree of the polyvinyl alcohol is 99 percent, and the average polymerization degree is 1700.
Sodium cyanoborohydride acts as a reducing agent to reduce imino groups to amino groups; the addition after completion of the schiff base reaction is because an imino group is generated only after the completion of the reaction, and no imino group is generated before the reaction, so that the reduction of an imino group is performed by adding sodium cyanoborohydride after the completion of the schiff base reaction.
Glutaraldehyde is a cross-linking agent in the reaction, and the carboxymethyl chitosan and the polyethyleneimine are cross-linked together through glutaraldehyde at normal temperature.
The polyvinyl alcohol has the functions of: a dispersant.
And if the usage amount of the polyethyleneimine is different, the functionalized carboxymethyl chitosan composite adsorbent with different polyethyleneimine grafting amounts can be finally obtained.
The functionalized carboxymethyl chitosan composite adsorbent of the invention has the following static adsorption and desorption processes to 0.5mg/mL phenylethanoid glycoside solution:
step 1, preparing a phenylethanoid glycoside solution:
immersing cistanche tubulosa sample (18-22mg) in 500-520mL 50% (v/v) ethanol solution, and carrying out ultrasonic treatment for 1.5-2 h. The residue in the solution was separated by centrifugation at 5000-. The centrifuged supernatant is passed through a 0.22-0.45 μm filter in sequence. The filtrate was then concentrated and dried in a vacuum oven. Finally, the dried product was dissolved in water to give a phenylethanoid glycoside solution with a concentration of 0.5 mg/mL.
Step 2, static adsorption and desorption process:
weighing 10-15mg of functionalized carboxymethyl chitosan composite adsorbent into a 50mL conical flask, adding 15-20mL of phenethyl alcohol glycoside solution with the concentration of 0.5mg/mL, sealing and placing on a 150-rpm shaker, oscillating at 30 ℃ for 12-24h, taking supernatant, and detecting the content of echinacoside and verbascoside by HPLC (high performance liquid chromatography), thereby calculating the adsorption amount and the adsorption rate of the functionalized carboxymethyl chitosan composite adsorbent on the echinacoside and the verbascoside. The adsorption amount of echinacoside is 28.761-347.975mg/g, the adsorption amount of verbascoside is 90.425-641.883mg/g, and the selectivity is 1.83-3.14. The adsorption rates were respectively: 3.8-45.6% of echinacoside and 11.9-84.4% of verbascoside. And then placing the fully adsorbed functionalized carboxymethyl chitosan composite adsorbent into a 50mL conical flask, adding an eluent (methanol: acetic acid: water: 1:8, v: v: v) to elute for 6-8h under the oscillation of 150-200rpm at 45 ℃, and then placing the obtained eluted functionalized carboxymethyl chitosan composite adsorbent into a vacuum drying oven to dry for 12-24h at 40 ℃. HPLC is adopted to detect the adsorption quantity of echinacoside and verbascoside in the extract. It was found that echinacoside and verbascoside in the adsorbent had been eluted.
After elution, the functionalized carboxymethyl chitosan composite adsorbent can adsorb echinacoside and verbascoside again, and can be repeatedly used.
And adsorbing the echinacoside and verbascoside again by the eluted functional carboxymethyl chitosan composite adsorbent, detecting the content of the echinacoside and the verbascoside by adopting HPLC (high performance liquid chromatography), and calculating to obtain the adsorption quantity and the adsorption rate of the functional carboxymethyl chitosan composite adsorbent for adsorbing the echinacoside and the verbascoside again. The re-adsorption amount of echinacoside reaches 21.483-312.339mg/g, the re-adsorption amount of verbascoside reaches 80.335-601.576mg/g, and the selectivity is 1.93-3.74. The re-adsorption rates were respectively: echinacoside 2.6-38.1%, verbascoside 10.3-77.2%.
Example 1
The preparation method of the functionalized carboxymethyl chitosan composite adsorbent comprises the following steps:
1.0g of carboxymethyl chitosan, 1.5g of a 50 wt% polyethyleneimine aqueous solution and 0.5g of polyvinyl alcohol were dissolved in 120mL of ultrapure water, 20mL of a 5 wt% glutaraldehyde aqueous solution was added dropwise to the solution (dropwise addition was completed for 30min), and the mixture was vigorously stirred at 25 ℃ for 6 hours (completion of Schiff base reaction) to obtain a pink flocculent precipitate. After 6h, sodium cyanoborohydride was added to the reaction, and 60mg was added every 2h for a total of 3 times. After the reaction is finished, washing the reaction product by deionized water for a plurality of times, and freezing the reaction product in vacuum for 24 hours to obtain the functionalized carboxymethyl chitosan composite adsorbent.
Wherein, the substitution degree of the carboxymethyl chitosan is 80 percent.
The molecular weight of the polyethyleneimine is 70000.
The hydrolysis degree of the polyvinyl alcohol is 99 percent, and the average polymerization degree is 1700.
The functionalized carboxymethyl chitosan composite adsorbent of the invention has the following static adsorption and desorption processes to 0.5mg/mL phenylethanoid glycoside solution:
step 1, preparing a phenylethanoid glycoside solution:
20mg of cistanche tubulosa sample is immersed in 500mL of 50% (v/v) ethanol solution and is subjected to ultrasonic treatment for 2 h. The residue in the solution was separated by centrifugation at 5000rpm for 10 min. The supernatant after centrifugation was passed through a 0.22 μm filter in that order. The filtrate was then concentrated and dried in a vacuum oven. Finally, the dried product was dissolved in water to give a phenylethanoid glycoside solution with a concentration of 0.5 mg/mL.
Step 2, static adsorption and desorption process:
weighing 10mg of functionalized carboxymethyl chitosan composite adsorbent into a 50mL conical flask, adding 15mL of phenylethanoid glycoside solution with the concentration of 0.5mg/mL, sealing and placing on a shaker at 150rpm, oscillating at 30 ℃ for 24h, taking supernatant, and detecting the contents of echinacoside and verbascoside by HPLC (high performance liquid chromatography), thereby calculating the adsorption amount and the adsorption rate of the functionalized carboxymethyl chitosan composite adsorbent on the echinacoside and the verbascoside. The adsorption amount of echinacoside reaches 35.891mg/g, the adsorption amount of verbascoside reaches 108.892mg/g, and the selectivity is 3.03. The adsorption rates were respectively: echinacoside 4.7%, verbascoside 14.3%. Then, the fully adsorbed functionalized carboxymethyl chitosan composite adsorbent is placed in a 50mL conical flask, eluent (methanol: acetic acid: water: 1:8, v: v: v) is added to elute for 6h at 45 ℃ under the shaking of 150rpm, and then the obtained eluted functionalized carboxymethyl chitosan composite adsorbent is placed in a vacuum drying oven and dried for 12h at 40 ℃.
And adsorbing the echinacoside and verbascoside again by the eluted functional carboxymethyl chitosan composite adsorbent, detecting the content of the echinacoside and the verbascoside by adopting HPLC (high performance liquid chromatography), and calculating to obtain the adsorption quantity and the adsorption rate of the functional carboxymethyl chitosan composite adsorbent for adsorbing the echinacoside and the verbascoside again. The re-adsorption quantity of echinacoside reaches 30.337mg/g, the re-adsorption quantity of verbascoside reaches 98.215mg/g, and the selectivity is 3.24. The re-adsorption rates were respectively: echinacoside 3.7%, verbascoside 12.6%.
Example 2
The preparation method of the functionalized carboxymethyl chitosan composite adsorbent of the embodiment is different from that of the embodiment 1 in that: the amount of polyethyleneimine added was 3 g. The rest steps are the same.
The static adsorption and desorption process of the functionalized carboxymethyl chitosan composite adsorbent for 0.5mg/mL phenylethanoid glycoside solution in the embodiment is the same as that in the embodiment 1.
The adsorption results were: the adsorption amount of echinacoside reaches 215.515mg/g, the adsorption amount of verbascoside reaches 475.931mg/g, and the selectivity is 2.21. The adsorption rates were respectively: echinacoside 28.3%, verbascoside 62.6%.
And adsorbing the echinacoside and verbascoside again by the eluted functional carboxymethyl chitosan composite adsorbent, detecting the content of the echinacoside and the verbascoside by adopting HPLC (high performance liquid chromatography), and calculating to obtain the adsorption quantity and the adsorption rate of the functional carboxymethyl chitosan composite adsorbent for adsorbing the echinacoside and the verbascoside again. The re-adsorption amount of echinacoside reaches 197.488mg/g, the re-adsorption amount of verbascoside reaches 453.308mg/g, and the selectivity is 2.30. The re-adsorption rates were respectively: echinacoside 24.1%, verbascoside 58.3%.
Example 3
The preparation method of the functionalized carboxymethyl chitosan composite adsorbent of the embodiment is different from that of the embodiment 1 in that: the amount of polyethyleneimine added was 4.5 g. The rest steps are the same.
The static adsorption and desorption process of the functionalized carboxymethyl chitosan composite adsorbent for 0.5mg/mL phenylethanoid glycoside solution in the embodiment is the same as that in the embodiment 1.
The adsorption results were: the adsorption amount of echinacoside reaches 347.975mg/g, the adsorption amount of verbascoside reaches 641.883mg/g, and the selectivity is 1.84. The adsorption rates were respectively: echinacoside 45.6%, verbascoside 84.4%.
And adsorbing the echinacoside and verbascoside again by the eluted functional carboxymethyl chitosan composite adsorbent, detecting the content of the echinacoside and the verbascoside by adopting HPLC (high performance liquid chromatography), and calculating to obtain the adsorption quantity and the adsorption rate of the functional carboxymethyl chitosan composite adsorbent for adsorbing the echinacoside and the verbascoside again. The re-adsorption quantity of echinacoside reaches 312.339mg/g, the re-adsorption quantity of verbascoside reaches 601.576mg/g, and the selectivity is 1.93. The re-adsorption rates were respectively: echinacoside 38.1%, verbascoside 77.2%.
Example 4
The preparation method of the functionalized carboxymethyl chitosan composite adsorbent of the embodiment is different from that of the embodiment 1 in that: the amount of polyethyleneimine added was 6 g. The rest steps are the same.
The static adsorption and desorption process of the functionalized carboxymethyl chitosan composite adsorbent for 0.5mg/mL phenylethanoid glycoside solution in the embodiment is the same as that in the embodiment 1.
The adsorption results were: the adsorption amount of echinacoside reaches 338.007mg/g, the adsorption amount of verbascoside reaches 620.031mg/g, and the selectivity is 1.83. The adsorption rates were respectively: echinacoside 44.3%, verbascoside 81.5%.
And adsorbing the echinacoside and verbascoside again by the eluted functional carboxymethyl chitosan composite adsorbent, detecting the content of the echinacoside and the verbascoside by adopting HPLC (high performance liquid chromatography), and calculating to obtain the adsorption quantity and the adsorption rate of the functional carboxymethyl chitosan composite adsorbent for adsorbing the echinacoside and the verbascoside again. The re-adsorption quantity of echinacoside reaches 300.221mg/g, the re-adsorption quantity of verbascoside reaches 584.108mg/g, and the selectivity is 1.95. The re-adsorption rates were respectively: echinacoside 36.6%, verbascoside 74.9%.
Comparative example 1
Preparing the functional carboxymethyl chitosan composite adsorbent without reducing imino. 1.0g of carboxymethyl chitosan, 4.5g of polyethyleneimine, 0.5g of polyvinyl alcohol were dissolved in 120mL of ultrapure water, and 20mL of an aqueous glutaraldehyde solution was added dropwise to the solution (30min), followed by vigorous stirring at 25 ℃ for 6h (completion of the Schiff base reaction), to obtain a pink flocculent precipitate. After the reaction is finished, washing the reaction product for a plurality of times by using deionized water, and freezing the reaction product for 24 hours in vacuum to obtain the functional carboxymethyl chitosan composite adsorbent without reducing imino groups. Calculated after adsorption according to the method of example 1, the echinacoside adsorption was 257.945mg/g, verbascoside adsorption was 574.194mg/g, and selectivity was 2.23. The adsorption rates were respectively: echinacoside 33.8%, verbascoside 75.5%.
And adsorbing the echinacoside and verbascoside again by the eluted functional carboxymethyl chitosan composite adsorbent, detecting the content of the echinacoside and the verbascoside by adopting HPLC (high performance liquid chromatography), and calculating to obtain the adsorption quantity and the adsorption rate of the functional carboxymethyl chitosan composite adsorbent for adsorbing the echinacoside and the verbascoside again. The re-adsorption quantity of echinacoside reaches 213.804mg/g, the re-adsorption quantity of verbascoside reaches 533.661mg/g, and the selectivity is 2.50. The re-adsorption rates were respectively: echinacoside 26.1%, verbascoside 68.5%.
As can be seen from comparative example 1, if the amount of polyethyleneimine is too small, the adsorption effect of the composite adsorbent prepared by the reduced imino method is not as good as that of the composite adsorbent prepared by the non-reduced imino method, although the amount of polyethyleneimine is too large. It can be seen that the amount of polyethyleneimine is very important.
Example 5
The preparation method of the functionalized carboxymethyl chitosan composite adsorbent comprises the following steps:
1g of carboxymethyl chitosan, 1.5g of a 50 wt% polyethyleneimine aqueous solution and 0.4g of polyvinyl alcohol were dissolved in 130mL of ultrapure water, 15mL of a 5 wt% glutaraldehyde aqueous solution was added dropwise to the solution (dropwise addition was completed within 30min), and the mixture was vigorously stirred at room temperature for 7 hours (completion of Schiff base reaction) to obtain a pink flocculent precipitate. After 7h, sodium cyanoborohydride was added to the reaction, 40mg every 3h, 3 times in total. After the reaction is finished, washing with deionized water for several times, and freezing in vacuum for 20h to obtain the functionalized carboxymethyl chitosan composite adsorbent.
Wherein, the substitution degree of the carboxymethyl chitosan is 83 percent.
The molecular weight of the polyethyleneimine is 1800.
The hydrolysis degree of the polyvinyl alcohol is 99 percent, and the average polymerization degree is 1700.
The static adsorption and desorption process of the functionalized carboxymethyl chitosan composite adsorbent on 0.5mg/mL phenylethanoid glycoside solution is as follows:
step 1, preparing a phenylethanoid glycoside solution:
20mg of cistanche tubulosa sample is immersed in 500mL of 50% (v/v) ethanol solution and is subjected to ultrasonic treatment for 2 h. The residue in the solution was separated by centrifugation at 5000rpm for 10 min. The supernatant after centrifugation was passed through a 0.22 μm filter in that order. The filtrate was then concentrated and dried in a vacuum oven. Finally, the dried product was dissolved in water to give a phenylethanoid glycoside solution with a concentration of 0.5 mg/mL.
Step 2, static adsorption and desorption process:
weighing 10mg of functionalized carboxymethyl chitosan composite adsorbent into a 50mL conical flask, adding 15mL of phenethyl alcohol glycoside solution with the concentration of 0.5mg/mL, sealing and placing on a 150rpm shaking table, oscillating at 30 ℃ for 24h, taking supernate, and detecting the content of echinacoside and verbascoside in the supernate by HPLC (high performance liquid chromatography), thereby calculating the adsorption quantity and the adsorption rate of the functionalized carboxymethyl chitosan composite adsorbent on the echinacoside and the verbascoside. The adsorption amount of echinacoside reaches 28.761mg/g, the adsorption amount of verbascoside reaches 90.425mg/g, and the selectivity is 3.14. The adsorption rates were respectively: echinacoside 3.8%, verbascoside 11.9%. Then, the fully adsorbed functionalized carboxymethyl chitosan composite adsorbent is placed in a 50mL conical flask, eluent (methanol: acetic acid: water: 1:8, v: v: v) is added to elute for 8h at 45 ℃ under the shaking of 150rpm, and then the obtained eluted functionalized carboxymethyl chitosan composite adsorbent is placed in a vacuum drying oven and dried for 12h at 40 ℃.
And adsorbing the echinacoside and verbascoside again by the eluted functional carboxymethyl chitosan composite adsorbent, detecting the content of the echinacoside and the verbascoside by adopting HPLC (high performance liquid chromatography), and calculating to obtain the adsorption quantity and the adsorption rate of the functional carboxymethyl chitosan composite adsorbent for adsorbing the echinacoside and the verbascoside again. The re-adsorption amount of echinacoside reaches 21.483mg/g, the re-adsorption amount of verbascoside reaches 80.335mg/g, and the selectivity is 3.74. The re-adsorption rates were respectively: echinacoside 2.6%, verbascoside 10.3%.
Example 6
The preparation method of the functionalized carboxymethyl chitosan composite adsorbent comprises the following steps:
1g of carboxymethyl chitosan, 6.0g of a 50 wt% polyethyleneimine aqueous solution and 0.6g of polyvinyl alcohol were dissolved in 110mL of ultrapure water, 20mL of a 5 wt% glutaraldehyde aqueous solution was added dropwise to the solution (dropwise addition was completed within 30min), and the mixture was vigorously stirred at room temperature for 5 hours (completion of Schiff base reaction), thereby obtaining a pink flocculent precipitate. After 5h, sodium cyanoborohydride was added to the reaction, and 60mg was added every 2h for a total of 3 times. After the reaction is finished, washing the reaction product with deionized water for a plurality of times, and freezing the reaction product in vacuum for 24 hours to obtain the functionalized carboxymethyl chitosan composite adsorbent.
Wherein, the substitution degree of the carboxymethyl chitosan is 81 percent.
The molecular weight of the polyethyleneimine is 20000.
The hydrolysis degree of the polyvinyl alcohol is 99 percent, and the average polymerization degree is 1700.
The functionalized carboxymethyl chitosan composite adsorbent of the invention has the following static adsorption and desorption processes to 0.5mg/mL phenylethanoid glycoside solution:
step 1, preparing a phenylethanoid glycoside solution:
18mg of cistanche tubulosa sample is immersed in 500mL of 50% (v/v) ethanol solution and is subjected to ultrasonic treatment for 2 h. The residue in the solution was separated by centrifugation at 6000rpm for 10 min. The supernatant after centrifugation was passed through a 0.22 μm filter in that order. The filtrate was then concentrated and dried in a vacuum oven. Finally, the dried product was dissolved in water to give a phenylethanoid glycoside solution with a concentration of 0.5 mg/mL.
Step 2, static adsorption and desorption process:
weighing 15mg of functionalized carboxymethyl chitosan composite adsorbent into a 50mL conical flask, adding 20mL of phenethyl alcohol glycoside solution with the concentration of 0.5mg/mL, sealing and placing on a shaker with 200rpm, oscillating at 30 ℃ for 12h, taking supernatant, detecting the content of echinacoside and verbascoside in the supernatant by HPLC (high performance liquid chromatography), and calculating the adsorption amount and the adsorption rate of the functionalized carboxymethyl chitosan composite adsorbent on the echinacoside and the verbascoside. The adsorption amount of echinacoside reaches 306.125mg/g, the adsorption amount of verbascoside reaches 593.437mg/g, and the selectivity is 1.94. The adsorption rates were respectively: echinacoside 40.15%, verbascoside 78.05%. Then, the fully adsorbed functionalized carboxymethyl chitosan composite adsorbent is placed in a 50mL conical flask, eluent (methanol: acetic acid: water: 1:8, v: v: v) is added to elute for 6h at 45 ℃ and 200rpm under shaking, and then the obtained eluted functionalized carboxymethyl chitosan composite adsorbent is placed in a vacuum drying oven and dried for 24h at 40 ℃.
And adsorbing the echinacoside and verbascoside again by the eluted functional carboxymethyl chitosan composite adsorbent, detecting the content of the echinacoside and the verbascoside by adopting HPLC (high performance liquid chromatography), and calculating to obtain the adsorption quantity and the adsorption rate of the functional carboxymethyl chitosan composite adsorbent for adsorbing the echinacoside and the verbascoside again. The re-adsorption quantity of echinacoside reaches 278.117mg/g, the re-adsorption quantity of verbascoside reaches 554.185mg/g, and the selectivity is 1.99. The re-adsorption rates were respectively: echinacoside 34.7%, verbascoside 71.3%.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (11)
1. The application of a functionalized carboxymethyl chitosan composite adsorbent in efficiently separating phenylethanoid glycosides compounds in cistanche tubulosa; the phenylethanoid glycosides compounds are echinacoside and verbascoside; the preparation method of the adsorbent comprises the following steps:
carboxymethyl chitosan and polyethyleneimine are used as raw materials, a dispersant and a cross-linking agent are added to perform Schiff base reaction to obtain pink flocculent precipitate, a reducing agent is added to the pink flocculent precipitate to reduce imino into amino, and the functionalized carboxymethyl chitosan composite adsorbent is obtained.
2. Use according to claim 1, characterized in that: the application specifically comprises the following steps: adding a functionalized carboxymethyl chitosan composite adsorbent into the phenylethanoid glycoside solution, and fully oscillating for adsorption.
3. Use according to claim 2, characterized in that: the adsorption conditions are as follows: 150 ℃ and 200rpm, and the adsorption is carried out for 12-24h under 30 ℃ oscillation.
4. Use according to claim 1, characterized in that: the functionalized carboxymethyl chitosan composite adsorbent is eluted after adsorption, and then the phenylethanol glycoside compounds are adsorbed again; the elution conditions were: the eluent is: methanol: acetic acid: the volume ratio of water is 1:1: 8; elution was carried out at 45 ℃ for 6-8h with shaking at 150 and 200 rpm.
5. Use according to any one of claims 1 to 4, characterized in that: the weight ratio of the carboxymethyl chitosan to the polyethyleneimine is 1: 1.5-6.
6. Use according to claim 5, characterized in that: the weight ratio of the carboxymethyl chitosan to the polyethyleneimine is 1: 4.5-6.
7. Use according to claim 6, characterized in that: the weight ratio of the carboxymethyl chitosan to the polyethyleneimine is 1: 4.5.
8. Use according to any one of claims 1 to 4, characterized in that: the dispersing agent is polyvinyl alcohol; and/or
The cross-linking agent is glutaraldehyde; and/or
The reducing agent is sodium cyanoborohydride; and/or
The schiff base reaction is completed at room temperature.
9. Use according to any one of claims 1 to 4, characterized in that: the degree of substitution of the carboxymethyl chitosan is not less than 80%.
10. Use according to any one of claims 1 to 4, characterized in that: the molecular weight of the polyethyleneimine is 1800-.
11. Use according to claim 8, characterized in that: the hydrolysis degree of the polyvinyl alcohol is 99%, and the average polymerization degree is 1700.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011383247.8A CN112604672B (en) | 2020-12-01 | 2020-12-01 | Functionalized carboxymethyl chitosan composite adsorbent and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011383247.8A CN112604672B (en) | 2020-12-01 | 2020-12-01 | Functionalized carboxymethyl chitosan composite adsorbent and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112604672A CN112604672A (en) | 2021-04-06 |
| CN112604672B true CN112604672B (en) | 2022-08-16 |
Family
ID=75228361
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011383247.8A Active CN112604672B (en) | 2020-12-01 | 2020-12-01 | Functionalized carboxymethyl chitosan composite adsorbent and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112604672B (en) |
Citations (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102744044A (en) * | 2012-07-31 | 2012-10-24 | 武汉大学 | Amino-protected crosslinking magnetic chitosan composite adsorbent and preparation method thereof |
| CN103469362A (en) * | 2013-09-10 | 2013-12-25 | 中山大学 | Porous polyethyleneimine and chitosan blend fiber and preparation method and applications thereof |
| CN104258830A (en) * | 2014-10-14 | 2015-01-07 | 河北工业大学 | Polyethyleneimine modified chitosan microsphere medium as well as preparation and application methods thereof |
| CN104549174A (en) * | 2013-10-10 | 2015-04-29 | 中国科学院大连化学物理研究所 | Phenylboronic acid-modified polyethyleneimine hybrid magnetic nanoparticle and its preparation method and use |
| CN105413661A (en) * | 2015-12-29 | 2016-03-23 | 华东理工大学 | Polyethyleneimine-chitosan adsorbent one-step synthesis method |
| CN105854839A (en) * | 2016-04-15 | 2016-08-17 | 武汉轻工大学 | Enzymatic crosslinking chitosan/polyethyleneimine grafted magnetic gelatin material and preparing method and application thereof |
| CN108003204A (en) * | 2017-11-09 | 2018-05-08 | 石河子大学 | A kind of method and its application that benzyl carbinol glycoside material is extracted from Cistanche tubulosa |
| CN107998183A (en) * | 2017-12-26 | 2018-05-08 | 浙江大学 | Sweet osmanthus benzyl carbinol glycosides liposome of Chitosan-coated and preparation method thereof |
| CN108298629A (en) * | 2018-01-09 | 2018-07-20 | 中国科学院新疆理化技术研究所 | A kind of preparation method of the composite material of efficient absorption mercury ion |
| CN108421534A (en) * | 2018-02-08 | 2018-08-21 | 中山大学 | A kind of chitosan gel rubber material and preparation method thereof, wastewater treatment method and application |
| CN108912185A (en) * | 2018-09-21 | 2018-11-30 | 罗永恒 | A kind of method and application preparing benzyl carbinol glycosides using Aricennia marina fruits |
| CN109674805A (en) * | 2019-02-12 | 2019-04-26 | 大连大学 | Application of the benzyl carbinol glycoside compound in the drug for promoting periodontosis Bone Defect Repari |
| CN110016063A (en) * | 2019-03-27 | 2019-07-16 | 石河子大学 | A kind of method and application based on benzyl carbinol glycosides in mesoporous carbon sublimated Cistanche tubulosa |
| WO2020014610A1 (en) * | 2018-07-12 | 2020-01-16 | International Flavors & Fragrances Inc. | Verbascoside and related compounds for sweetness enhancement |
| CN110862568A (en) * | 2019-12-04 | 2020-03-06 | 石河子大学 | Preparation method and application of molecularly imprinted membrane for separating phenylethanoid glycosides |
-
2020
- 2020-12-01 CN CN202011383247.8A patent/CN112604672B/en active Active
Patent Citations (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102744044A (en) * | 2012-07-31 | 2012-10-24 | 武汉大学 | Amino-protected crosslinking magnetic chitosan composite adsorbent and preparation method thereof |
| CN103469362A (en) * | 2013-09-10 | 2013-12-25 | 中山大学 | Porous polyethyleneimine and chitosan blend fiber and preparation method and applications thereof |
| CN104549174A (en) * | 2013-10-10 | 2015-04-29 | 中国科学院大连化学物理研究所 | Phenylboronic acid-modified polyethyleneimine hybrid magnetic nanoparticle and its preparation method and use |
| CN104258830A (en) * | 2014-10-14 | 2015-01-07 | 河北工业大学 | Polyethyleneimine modified chitosan microsphere medium as well as preparation and application methods thereof |
| CN105413661A (en) * | 2015-12-29 | 2016-03-23 | 华东理工大学 | Polyethyleneimine-chitosan adsorbent one-step synthesis method |
| CN105854839A (en) * | 2016-04-15 | 2016-08-17 | 武汉轻工大学 | Enzymatic crosslinking chitosan/polyethyleneimine grafted magnetic gelatin material and preparing method and application thereof |
| CN108003204A (en) * | 2017-11-09 | 2018-05-08 | 石河子大学 | A kind of method and its application that benzyl carbinol glycoside material is extracted from Cistanche tubulosa |
| CN107998183A (en) * | 2017-12-26 | 2018-05-08 | 浙江大学 | Sweet osmanthus benzyl carbinol glycosides liposome of Chitosan-coated and preparation method thereof |
| CN108298629A (en) * | 2018-01-09 | 2018-07-20 | 中国科学院新疆理化技术研究所 | A kind of preparation method of the composite material of efficient absorption mercury ion |
| CN108421534A (en) * | 2018-02-08 | 2018-08-21 | 中山大学 | A kind of chitosan gel rubber material and preparation method thereof, wastewater treatment method and application |
| WO2020014610A1 (en) * | 2018-07-12 | 2020-01-16 | International Flavors & Fragrances Inc. | Verbascoside and related compounds for sweetness enhancement |
| CN108912185A (en) * | 2018-09-21 | 2018-11-30 | 罗永恒 | A kind of method and application preparing benzyl carbinol glycosides using Aricennia marina fruits |
| CN109674805A (en) * | 2019-02-12 | 2019-04-26 | 大连大学 | Application of the benzyl carbinol glycoside compound in the drug for promoting periodontosis Bone Defect Repari |
| CN110016063A (en) * | 2019-03-27 | 2019-07-16 | 石河子大学 | A kind of method and application based on benzyl carbinol glycosides in mesoporous carbon sublimated Cistanche tubulosa |
| CN110862568A (en) * | 2019-12-04 | 2020-03-06 | 石河子大学 | Preparation method and application of molecularly imprinted membrane for separating phenylethanoid glycosides |
Non-Patent Citations (2)
| Title |
|---|
| "Highly efficient and selective removal of mercury ions using hyperbranched polyethylenimine functionalized carboxymethyl chitosan composite adsorbent";Zeng, HH et al.;《Chemical Engineering Journal》;20181002;第358卷;第254页右栏第2.2节，附加信息第2页2.1节 * |
| "壳聚糖化学改性新进展";赵希荣;《精细与专用化学品》;20040806(第18期);第19-20页第4节 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112604672A (en) | 2021-04-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113648834A (en) | Ceramic membrane and preparation method and application thereof | |
| CN103058871B (en) | Separation and purification method of tobacco chlorogenic acid | |
| CN112920034A (en) | Method for extracting 6-gingerol with content of more than or equal to 98% | |
| CN112159747A (en) | Method for removing fusel oil in yellow wine | |
| JP6703562B2 (en) | Method for producing polyphenol composition from bagasse | |
| CN112604672B (en) | Functionalized carboxymethyl chitosan composite adsorbent and preparation method and application thereof | |
| CN112321656B (en) | Method for separating and preparing acylated anthocyanin | |
| CN112175028A (en) | Method for separating and preparing delphinidin-3-O- (6-O-p-coumaroyl) glucoside | |
| CN116891559A (en) | Preparation method of organic framework material and method for extracting total chlorogenic acid | |
| CN106542999A (en) | A kind of purification process of rosmarinic acid | |
| US8766019B2 (en) | Method for separating butanol | |
| CN113214531B (en) | Hydrophobic/hydrophilic interpenetrating network resin and preparation and application thereof | |
| CN112645994B (en) | Extraction process of salidroside | |
| CN108014176B (en) | Method for purifying guava leaf triterpenes | |
| CN108745315B (en) | Cellulose-modified chloromethylated polystyrene hypercrosslinked resin and adsorption separation method | |
| CN114057753B (en) | Method for separating and purifying antifungal active substance HSAF in zymogen fermentation liquor | |
| CN107760057A (en) | A kind of Suaeda salsa Betacyanins extracting method | |
| CN113801134A (en) | Production process for simultaneously producing ginkgolide, ginkgetin, ginkgopolysaccharide and shikimic acid | |
| CN103408614A (en) | Novel preparation technique of steviosin and Rebaudioside-A | |
| CN109400743B (en) | Method for extracting sulfated galactan from sea grapes | |
| CN111978366A (en) | Method for extracting dioscin from fenugreek | |
| CN111374993A (en) | Method for extracting acanthopanax senticosus total saponin from acanthopanax senticosus leaves | |
| CN101974014B (en) | Manufacturing technology for extracting ginkalide A and C from root and bark of maidenhair tree | |
| CN111714570A (en) | Separation and purification method of pineapple peel flavone and flavone | |
| CN110669172A (en) | Preparation method and application of molecularly imprinted polymer |
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 |