CN107880008B - Method for extracting luteolin - Google Patents
Method for extracting luteolin Download PDFInfo
- Publication number
- CN107880008B CN107880008B CN201711115071.6A CN201711115071A CN107880008B CN 107880008 B CN107880008 B CN 107880008B CN 201711115071 A CN201711115071 A CN 201711115071A CN 107880008 B CN107880008 B CN 107880008B
- Authority
- CN
- China
- Prior art keywords
- magnetic nanoparticles
- magnetic
- eluent
- sonication
- mnp
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/28—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only
- C07D311/30—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3 with aromatic rings attached in position 2 only not hydrogenated in the hetero ring, e.g. flavones
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/22—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4
- C07D311/26—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 4 with aromatic rings attached in position 2 or 3
- C07D311/40—Separation, e.g. from natural material; Purification
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The embodiment of the invention provides a method for extracting luteolinThe method comprises the following steps: (1) obtaining the extract containing luteolin and Fe3O4Magnetic nanoparticles of said Fe3O4Modifying the magnetic nano particles by amino; (2) mixing the solution to be extracted and the Fe3O4Mixing magnetic nano particles, and carrying out first ultrasonic treatment; (3) after the first ultrasonic treatment, Fe is treated under the action of an external magnetic field3O4Separating out the magnetic nano particles; (4) and (3) separating Fe separated in the step (3)3O4Mixing the magnetic nanoparticles with the eluent, and carrying out secondary ultrasonic treatment; the eluent comprises glacial acetic acid and methanol aqueous solution; (5) after the second ultrasonic treatment, Fe is treated under the action of an external magnetic field3O4Separating the magnetic nanoparticles to obtain an eluate containing luteolin.
Description
Technical Field
The invention relates to the technical field of compound extraction, in particular to a method for extracting luteolin.
Background
With the rapid development of nano science, magnetic nano materials get more and more attention in different fields. Difference between magnetic nano material and common magnetic materialThe main expression is in two aspects: the difference in size on the one hand and the surface area on the other hand results in different physical and other properties of the two magnetic materials. There are many kinds of magnetic nanomaterials, among which Fe3O4Magnetic nanoparticles are of great interest because of the advantages of relatively simple and convenient preparation process, low price, low toxicity, predictability and controllability of structure and function, and the like. Fe3O4The nanoparticles have a nano-size effect when Fe3O4When the particle size is less than 20nm, superparamagnetism is shown at normal temperature; the functional group which is easy to modify can generate specific affinity extraction with a target object; under the directional control of an external magnetic field, the target can be quickly separated from a multi-component environment through cleaning and desorption operations. Fe3O4Magnetic nanoparticles also have some disadvantages that cannot be overcome by themselves, bare Fe3O4Magnetic nanoparticles are very easy to oxidize in air and are easy to corrode and agglomerate in an acidic environment, so that the extraction effect and the extraction selectivity of the magnetic nanoparticles are poor. For increasing Fe3O4The magnetic nanoparticles have extraction performance, and can improve target capture ability, and multiple functional groups such as sulfhydryl (-SH), amino (-NH)2) Carboxyl (-COOH), sulfonic acid (-SO)3H) Hydroxyl (-OH), etc. are used for Fe3O4Modification of magnetic nanoparticles. The active groups have stronger chelation on heavy metal ions and are modified by Fe3O4The magnetic nano-particles can effectively and selectively extract a target object, so that the target object is more easily combined with a guest molecule, and the application of the magnetic nano-particles in the aspect of selective extraction of compounds can be increased. Although the unique characteristics of magnetic nanoparticles have attracted extensive attention in the field of biomedical science in the development of nanotechnology over the past decades, their application in the extraction of compounds in foods and Chinese medicines is still rare.
Disclosure of Invention
The inventors are on Fe3O4The intensive research on magnetic nanoparticles has led to the unexpected discovery that amino-modified Fe3O4The magnetic nanoparticles can be used for the extraction of luteolin and, based thereon,the present invention has been completed.
The invention provides a method for extracting luteolin, which comprises the following steps:
(1) obtaining the extract containing luteolin and Fe3O4Magnetic nanoparticles of said Fe3O4Modifying the magnetic nano particles by amino;
(2) mixing the solution to be extracted and the Fe3O4Mixing magnetic nano particles, and carrying out first ultrasonic treatment;
(3) after the first ultrasonic treatment, Fe is treated under the action of an external magnetic field3O4Separating out the magnetic nano particles;
(4) and (3) separating Fe separated in the step (3)3O4Mixing the magnetic nanoparticles with the eluent, and carrying out secondary ultrasonic treatment; the eluent comprises glacial acetic acid and methanol aqueous solution;
(5) after the second ultrasonic treatment, Fe is treated under the action of an external magnetic field3O4Separating the magnetic nanoparticles to obtain an eluate containing luteolin.
In the present invention, amino-modified Fe is used3O4The magnetic nanoparticles can be obtained by existing synthesis methods, here the invention of amino-modified Fe3O4The method of preparing the magnetic nanoparticles is not limited.
In a specific embodiment of the invention, the ultrasonic temperature of the first ultrasonic treatment is 0-45 ℃, and the ultrasonic time is 10-60 minutes.
In a specific embodiment of the invention, the ultrasonic temperature of the first ultrasonic treatment is 25-40 ℃, and the ultrasonic time is 25-45 minutes.
In one embodiment of the present invention, the first sonication is performed at a sonication temperature of 30 ℃ for a sonication time of 40 minutes.
In one embodiment of the present invention, Fe separated in step (3)3O4Fe before mixing the magnetic nanoparticles with the eluent3O4Magnetic nanoparticlesAnd (5) drying the granules. The drying treatment may be carried out by methods conventional in the art, for example by vacuum drying at 40-80 ℃ to constant weight.
In one embodiment of the present invention, in step (2), Fe3O4The ratio of the mass of the magnetic nanoparticles to the volume of the liquid to be extracted is greater than or equal to (5: 1) mg/mL.
In one embodiment of the present invention, in step (2), Fe3O4The ratio of the mass of the magnetic nanoparticles to the volume of the liquid to be extracted was (20: 1) mg/mL.
In one embodiment of the present invention, in step (4), Fe3O4The volume ratio of the mass of the magnetic nanoparticles to the eluent is 1: (1-4) mg/mL, preferably 1: 2.5 mg/mL.
In one embodiment of the present invention, in step (4), the time of the second ultrasonic treatment is 15 to 45 minutes, preferably 40 minutes.
In one embodiment of the present invention, in the step (4), the volume percentage concentration of glacial acetic acid in the eluent is 10% -30% based on the total volume of the eluent; preferably 20%.
In one embodiment of the present invention, in the step (4), the volume percentage concentration of methanol in the methanol aqueous solution is 40% to 80%; preferably 60%.
In a specific embodiment of the present invention, the liquid to be extracted is a plant liquid.
At present, the development and utilization of plant peanuts are mainly to develop peanut kernels and peanut red skins in the plant peanuts; most peanut hulls are discarded or used as fuel. Only a small amount is recycled, and part is processed into feed or used as chemical raw materials. The recycling of the production waste materials such as peanut shells and the like can improve the industrial production profit on the basis of greatly reducing the influence of industrial production on the environment and increase the comprehensive utilization value of economic crops. The peanut shell contains a large amount of luteolin, and the luteolin has good pharmacological activities of resisting oxidation, resisting cancer, resisting inflammation, inhibiting bacteria, reducing cholesterol and the like. The luteolin in the peanut shells is extracted and processed to prepare and refine the luteolin compounds or chemical derivative products thereof, the luteolin compounds are used for food and health products, and cheap industrial waste peanut shells can be transformed into renewable resources; becomes an ideal natural raw material of natural antioxidant and preservative. In one embodiment of the present invention, the extract to be extracted containing luteolin can be peanut shell extract.
The invention provides a method for extracting luteolin, which is characterized in that Fe modified by amino groups3O4The magnetic nanoparticles can be used as extractant for extracting luteolin from extractive solution containing luteolin into eluate, and the amino group modified Fe3O4The magnetic nanoparticles have high selectivity and good extraction efficiency on luteolin.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 shows MNP-NH2Scanning electron microscope images of;
FIG. 2 shows MNP-NH2Transmission electron microscopy images of (a);
FIG. 3 shows MNP-NH2And Fe3O4An infrared spectrum of (1);
FIG. 4 shows MNP-NH2And Fe3O4The hysteresis loop of (1).
FIG. 5 shows MNP-NH2A state diagram in the absence/presence of an external magnetic field after dispersion in water;
FIG. 6 shows MNP-NH of different qualities2The liquid chromatogram of the corresponding peanut shell extract is shown as 1-5 in the figure respectively: 1 is a blank group; 2 is 50mg MNP-NH2(ii) a3 is 100mg MNP-NH2(ii) a 4 is 150mg MNP-NH2(ii) a 5 is 200mg MNP-NH 2;
FIG. 7 shows different masses of MNP-NH2A liquid chromatogram of the corresponding eluent; 1-5 in the figure represent: 1 is a blank group; 2 is 200mg MNP-NH2(ii) a3 is 150mg MNP-NH2(ii) a 4 is 100mg MNP-NH2(ii) a 5 is 50mg MNP-NH2。
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
First, materials, instruments, and the like used in the following examples will be described.
1. Material
1.1 Experimental materials and reagents
Luteolin was purchased from Nanjing Zeron pharmaceutical company (purity > 98% by HPLC). The medicinal materials are as follows: flowers are produced in Hebei.
FeCl3·6H2O, 1, 6-hexanediamine, sodium acetate anhydrous (NaAc) available from Aladdin reagent Inc.; ethylene glycol, anhydrous ethanol, glacial acetic acid were purchased from Tianjin Kogyo reagent, Inc.; phosphoric acid was purchased from Fochen chemical reagent works, Tianjin; sodium phosphate, sodium hydroxide purchased from chemical reagent wholesale company, Tianjin; chromatography of formic acid, acetonitrile from Sigma, usa; the water is distilled water.
1.2 instruments
The main equipment used in the experiment is shown in table 1.
TABLE 1 Experimental instrumentation
Example 1 amino-modified Fe3O4Preparation and characterization of magnetic nanoparticles
1.1 amino-modified Fe3O4Magnetic nanoPreparation of rice granules
Preparation of MNP-NH by classical solvothermal synthesis2The method comprises the following specific steps: firstly FeCl3·6H2Adding O (2g) and 1, 6-hexanediamine (6g) into a 250mL round-bottom flask, adding 50mL of ethylene glycol, fully stirring and ultrasonically treating to dissolve blocky solids, adding NaAc (6g), and continuously and violently stirring for 30min to form uniform viscous liquid, wherein the color of the solution is gradually changed from light yellow to dark brown; transferring the mixed solution into a 50mL reaction kettle, sealing, putting into an oven, and reacting for 10h at 200 ℃; cooling to room temperature, separating out black solid with Nd-Fe-B magnet, sequentially and respectively ultrasonic washing with deionized water and anhydrous ethanol for 2 times, vacuum drying at 80 deg.C for 1h to obtain amino modified Fe3O4Magnetic nanoparticles (hereinafter abbreviated as MNP-NH)2)。
Non-aminated Fe for control in characterization3O4Preparation of
Firstly FeCl3·6H2Adding O (2g) into a 250mL round-bottom flask, adding 50mL ethylene glycol, fully stirring and ultrasonically treating to dissolve blocky solids, adding NaAc (6g), and continuously and violently stirring for 30min to form uniform viscous liquid; transferring the mixed solution into a 50mL reaction kettle, sealing, putting into an oven, and reacting for 10h at 200 ℃; cooling to room temperature, separating out black solid with Nd-Fe-B magnet, sequentially and respectively ultrasonic washing with deionized water and anhydrous ethanol for 2 times, vacuum drying at 80 deg.C for 1h to obtain magnetic Fe3O4。
1.2MNP-NH2Is characterized by
1.2.1 Electron microscopy analysis
The MNP-NH prepared under the section of '1.1' is determined by adopting a JSM 6700F type scanning electron microscope of JEOL company of Japan2The analysis of the topographic features of (a) is shown in fig. 1.
Adopting Hitachi H-800 high-resolution transmission electron microscope of Hitachi corporation of Japan, with vacuum degree greater than 10- 6Torr, photograph MNP-NH prepared under the term "1.12The results are shown in FIG. 2.
As can be seen from FIGS. 1 and 2, the MNP-NH was produced2The particle size of the MNP-NH is about 20nm, the distribution is uniform, the MNP-NH is basically a single layer and mainly has a quasi-spherical structure, and particles are locally aggregated and overlapped2。
1.2.2 Infrared Spectroscopy
Adopts a Bruker 550 type Fourier transform infrared spectrometer of Germany Bruker company, KBr tablets and the wave number range is 400-4000 cm-1Measuring the MNP-NH prepared as described above2Structural characterization of the chemical functional groups in the structure of (1), and Fe which has not been aminated3O4As a control; the results are shown in FIG. 3.
MNP-NH2See FIG. 3 (a), Fe3O4See fig. 3 (b); as can be seen from (a) and (b), 586cm-1Characteristic absorption peak of Fe-O-Fe bond; at 1650cm-1The absorption peak at (A) is due to the bending vibration of O-H, indicating that Fe3O4And MNP-NH2some-OH groups are present on the surface. (a) 3379cm in the figure-1And 1261cm-1The characteristic absorption of the magnetic particle is N-H strong stretching vibration absorption and C-N stretching vibration absorption of corresponding amino, which shows that amino functional groups are successfully modified on the surface of the magnetic particle after reaction.
1.2.3 magnetic assay analysis
Vibrating Sample Magnetometers (VSMs) are suitable for measurement of various magnetic materials. The experiment adopted superconducting quantum interference device (SQUID) of American Quantum design company to measure the prepared MNP-NH2Hysteresis loop of and with Fe3O4As a control; the results are shown in FIG. 4; wherein a in FIG. 4 is Fe3O4The hysteresis loop of (1); b is MNP-NH2The hysteresis loop of (1);
from FIG. 4, it is clear that MNP-NH can be found2Relative to Fe3O4The saturation magnetization is much reduced because of MNP-NH2The surface layer is modified with a certain amount of-NH2Has certain influence on the saturation magnetization degree, thereby reducing MNP-NH2Magnetic moment of (a). In addition, as can be seen from the above figures, when the intensity of the applied magnetic field is reduced to zero, the coercive force and remanence of the magnetic sample are both almost zero, and the hysteresis phenomenon is hardly observed, so that the magnetic nanoparticles are very suitable for being used as a carrier.
Alternatively, as shown in FIG. 5, MNP-NH prepared as described above2MNP-NH prepared after mixing with water in the absence of an externally applied magnetic field2Can be uniformly dispersed in the solution (see a picture in figure 5), and when an external magnetic field is applied (namely, when the magnet is close to the MNP), the MNP-NH can be uniformly dispersed in the solution2The polymer can be rapidly gathered in the direction of the magnetic field, and the uniform dispersion liquid rapidly generates a layering phenomenon (see a b picture in figure 5).
Example 2 optimization of extraction conditions
Using MNP-NH2When the luteolin is used as an extractant for extraction, the extraction process is divided into two parts, namely adsorption and elution. The adsorption part includes:
(2) mixing the solution to be extracted and the Fe3O4Mixing magnetic nano particles, and carrying out first ultrasonic treatment;
(3) after the first ultrasonic treatment, Fe is treated under the action of an external magnetic field3O4The magnetic nanoparticles are separated out.
The elution portion includes:
(4) and (3) separating Fe separated in the step (3)3O4Mixing the magnetic nanoparticles with the eluent, and carrying out secondary ultrasonic treatment; the eluent comprises glacial acetic acid and methanol aqueous solution;
(5) after the second ultrasonic treatment, Fe is treated under the action of an external magnetic field3O4Separating the magnetic nanoparticles to obtain an eluate containing luteolin.
In this embodiment, the peanut shell extract is used as the extract to be extracted, and MNP-NH is used2Extracting luteolin contained in the extract, and respectively optimizing the conditions related to the adsorption part and the elution part.
2.1 optimization of adsorption conditions
2.1.1 preparation of peanut Shell extract
Weighing 5g of peanut shell coarse powder, adding 100mL of 75% ethanol, performing ultrasonic treatment for 45 minutes, performing suction filtration, collecting filtrate, and storing at low temperature (5 ℃) for later use.
2.1.2 high Performance liquid chromatography conditions
A chromatographic column: waters Symmetry C18 (250X 4.6mm,5 μm); the mobile phase is acetonitrile (A) and 0.2 percent phosphoric acid water (B), gradient elution is carried out for 0-30 min, and the concentration of A is 15-40 percent; 30-31 min, 40% -15% A; 31-35 min, 15% -15% A; the detection wavelength is 254 nm. Flow rate: 1 mL. min-1(ii) a The sample volume is 10 mu L; column temperature: at 30 ℃.
2.1.3 examination of ultrasound temperature and ultrasound time for the first sonication
Taking 5mL of peanut shell extract prepared under the item of '2.1.1', adding 50mg of MNP-NH2Ultrasonic treatment at constant temperature of 25 ℃, and then MNP-NH is realized under the action of an external magnetic field (applied by a neodymium iron boron magnet)2Separating with peanut shell extractive solution (solid-liquid phase separation), centrifuging the separated peanut shell extractive solution at 20000r/min for 10min, collecting supernatant, passing through membrane, and performing high performance liquid detection under the chromatographic condition of "2.1.2"; taking out the MNP-NH2Performing high performance liquid detection on the peanut shell extracting solution after the same treatment; the non-MNP-NH addition was calculated by a standard curve method commonly used in the art2The concentration C of luteolin in the peanut shell extract0And after different ultrasonic time, with MNP-NH2The concentration C of luteolin remaining in the separated peanut shell extract1According to the formula Q ═ C [ (-)0-C1)×V1]Calculating the extraction amount of luteolin by using the/M, wherein Q represents the extraction amount; v1The volume of the peanut shell extract (5mL) is shown, and M represents MNP-NH2Mass of (2) (50 mg); and (3) observing the optimal ultrasonic time within the range of 0-50 min by calculating the extraction amount of luteolin.
The examination result shows that the extraction amount of the luteolin reaches the maximum value (saturated extraction amount) of 64mg/g after the ultrasonic time of the first ultrasonic treatment is 30 min. To ensure the stability of the experiment, the optimal first sonication time was finally selected to be 40min for further study.
Except that the first ultrasonic treatment time is fixed to be 40 minutes, the influence of the ultrasonic temperature on the extraction is researched by controlling the temperature to be 0-45 ℃ according to the investigation condition of the ultrasonic time of the first ultrasonic treatment.
The investigation result shows that the temperature has little influence on the extraction of luteolin, and finally 30 ℃ which is easier to control the constant temperature is selected as the optimal ultrasonic temperature.
2.2 optimization of elution conditions
The elution conditions were investigated using orthogonal experiments. Firstly, preliminarily determining that the eluent consists of glacial acetic acid and methanol aqueous solution; influencing MNP-NH2The main factors of the elution effect are: the volume percent concentration of glacial acetic acid in the eluent (A), the volume percent concentration of methanol in aqueous methanol (B), the elution time (C) and the elution volume (D). Using orthographic list L9 (3)4) A four-factor three-level orthogonal test was performed as shown in table 2. The experimental procedure was as follows:
adding 2mg MNP-NH into nine parts of 5mL peanut shell extractive solution prepared under 2.1.12Performing ultrasonic treatment at constant temperature of 30 deg.C for 40min, performing solid-liquid separation, centrifuging the separated peanut shell extractive solution at 20000r/min for 10min, collecting supernatant, passing through membrane, performing high performance liquid chromatography under the condition of chromatography of "2.1.2", and collecting the supernatant without MNP-NH2Performing high performance liquid detection on the peanut shell extracting solution after the same treatment; calculating the non-MNP-NH by adopting a standard curve method2The concentration C of luteolin in the peanut shell extract0And the concentration C of luteolin remaining in the separated peanut shell extract1。
Separating the MNP-NH2Drying at 80 deg.C, mixing with eluent, ultrasonic treating, and separating MNP-NH2Then, centrifuging the eluent at 20000r/min for 10min, taking the supernatant, passing through a membrane, and carrying out high performance liquid detection according to the chromatographic conditions under the item of 2.1.2; calculating luteolin concentration C in eluate by standard curve method2The luteolin elution rate is calculated according to the following formula:
luteolin elution rate ═ C2/(C0-C1)。
TABLE 2 MNP-NH2Elution effect orthogonal test design table
L9(34) The orthogonal test table and the luteolin elution rate are shown in Table 3. The analysis of the intuitive method shows that the factors influencing the elution rate of the luteolin are A from large to small>B>D (the factor C is taken as an error column), and the importance of each level in the factor A is A2 in sequence>A1>A3; the importance of the level among the B factors is B2 in turn>B3>B1; the importance of the level among the D factors is D2 in turn>D3>D1; therefore, the optimal condition is A2B2D 2; analysis of variance (table 4) showed that A, B and D were significantly different in the extraction yield factor for luteolin. The final elution conditions were 5mL of 20% glacial acetic acid (methanol: water: 60: 40), sonication 40min, combined with orthogonal experiments and analysis of variance. Namely: eluent composition: comprises 20% glacial acetic acid and 80% methanol water solution; the methanol volume percentage concentration of the methanol water solution is 60 percent, and the rest 40 percent is water; MNP-NH2The volume ratio of the mass to the eluent is 1: 2.5 mg/mL. The sonication time (i.e., elution time) for the second sonication was 40 minutes.
TABLE 3L 9 (3)4) Orthogonal test table and results
TABLE 4 luteolin elution Rate variance analysis Table
F1-0.05(2，2)＝19.000
2.3 peanut Shell extract and MNP-NH2Examination of the proportions
With MNP-NH2Taking 4 parts of 10mL peanut shell extract as extractant, and adding 50, 100, 150 and 200mg MNP-NH respectively2And carrying out ultrasonic treatment at 60 ℃ for 60 min. Under the action of an external magnetic fieldMNP-NH2Separating with peanut shell extractive solution (solid-liquid phase separation), centrifuging the separated peanut shell extractive solution at 20000r/min for 10min, collecting supernatant, passing through membrane, and performing high performance liquid detection under the chromatographic condition of "2.1.2"; and with no addition of MNP-NH2The peanut shell extract is used as a blank group control, and each high performance liquid chromatogram is shown in figure 6. Separated MNP-NH2Oven drying (80 deg.C), adding 10mL eluent, performing ultrasonic treatment at 30 deg.C for 60min, and separating MNP-NH2Then, centrifuging the eluent at 20000r/min for 10min, taking the supernatant, passing through a membrane, and carrying out high performance liquid detection according to the chromatographic conditions under the item of 2.1.2; the detection spectrum is shown in FIG. 7. Eluent composition: according to the volume percentage concentration: 20 percent of glacial acetic acid and 80 percent of methanol aqueous solution, wherein the volume percentage concentration of methanol in the methanol aqueous solution is 60 percent.
As can be seen from FIGS. 6 and 7, MNP-NH2The dosage and the proportion of the peanut shell extract are positively correlated with the extraction effect of luteolin, with MNP-NH2The ratio of the mass of (A) to the volume of the peanut shell extract increases, the adsorption amount increases, and the elution amount also increases. As can be seen from the figure, when MNP-NH2When the ratio of the mass of the extract to the volume of the peanut shell extract reaches (20: 1) mg/mL, the luteolin is extracted most; and MNP-NH2The extractant has selective extraction effect on luteolin in peanut shell, and has high extraction efficiency.
As can be seen from the above examples, MNP-NH prepared by the present invention2As can be seen by SEM, TEM, FT-IR and VSM characterization, the obtained MNP-NH2The magnetic separation method has good extraction capacity and strong superparamagnetism, is very easy to separate by adopting a magnetic separation method, and can be uniformly dispersed in a solution again under the condition of oscillation or ultrasound after an external magnetic field disappears, so that good cyclic utilization can be realized. The extract is suitable for extracting and separating luteolin from the extract containing luteolin. Compared with the traditional purification methods (such as precipitant method, extraction method, dialysis method, etc.), the method has the most remarkable advantages of strong selectivity and high efficiency in directly extracting luteolin from peanut shell extract. The whole separation process is time-saving and labor-saving, and the operation is carried outThe method is simple and easy to control. The method for separating and purifying the chemical components has the advantages of low energy consumption, environmental protection, good efficiency and low pollution, and provides a new idea and method for processing foods and Chinese medicinal preparations and recycling industrial waste.
It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.
All the embodiments in the present specification are described in a related manner, and the same and similar parts among the embodiments may be referred to each other, and each embodiment focuses on the differences from the other embodiments. In particular, for the system embodiment, since it is substantially similar to the method embodiment, the description is simple, and for the relevant points, reference may be made to the partial description of the method embodiment.
The above description is only for the preferred embodiment of the present invention, and is not intended to limit the scope of the present invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention shall fall within the protection scope of the present invention.
Claims (16)
1. A method for extracting luteolin is characterized by comprising the following steps:
(1) obtaining the extract containing luteolin and Fe3O4Magnetic nanoparticles of said Fe3O4Modifying the magnetic nano particles by amino; fe3O4The magnetic nanoparticles are made of FeCl3·6H2O, 1, 6-hexamethylene diamine and NaAc, and reacting for 10 hours at 200 ℃ to obtain the product;
(2) mixing the solution to be extracted and the Fe3O4Mixing magnetic nano particles, and carrying out first ultrasonic treatment;
(3) after the first ultrasonic treatment, Fe is treated under the action of an external magnetic field3O4Separating out the magnetic nano particles;
(4) and (3) separating Fe separated in the step (3)3O4Mixing the magnetic nanoparticles with the eluent, and carrying out secondary ultrasonic treatment; the eluent comprises glacial acetic acid and methanol aqueous solution;
(5) after the second ultrasonic treatment, Fe is treated under the action of an external magnetic field3O4Separating the magnetic nanoparticles to obtain an eluate containing luteolin.
2. The method of claim 1, wherein the first sonication is performed at a sonication temperature of 0 to 45 ℃ for a sonication time of 10 to 60 minutes.
3. The method of claim 1, wherein the first sonication is performed at a sonication temperature of 25 to 40 ℃ for a sonication time of 25 to 45 minutes.
4. The method of claim 1, wherein the first sonication is carried out at a sonication temperature of 30 ℃ and for a sonication time of 40 minutes.
5. The method of claim 1, wherein the Fe separated in step (3) is3O4Fe before mixing the magnetic nanoparticles with the eluent3O4And (5) drying the magnetic nanoparticles.
6. The method of claim 1, which isCharacterized in that, in step (2), Fe3O4The ratio of the mass of the magnetic nanoparticles to the volume of the liquid to be extracted is greater than or equal to (5: 1) mg/mL.
7. The method of claim 6, wherein in step (2), Fe3O4The ratio of the mass of the magnetic nanoparticles to the volume of the liquid to be extracted was (20: 1) mg/mL.
8. The method of claim 1, wherein in step (4), Fe3O4The volume ratio of the mass of the magnetic nanoparticles to the eluent is 1: (1-4) mg/mL.
9. The method of claim 1, wherein in step (4), Fe3O4The volume ratio of the mass of the magnetic nanoparticles to the eluent is 1: 2.5 mg/mL.
10. The method of claim 1, wherein in step (4), the second sonication time is between 15 and 45 minutes.
11. The method of claim 1, wherein in step (4), the second sonication time is 40 minutes.
12. The method of claim 1, wherein in step (4), the concentration of glacial acetic acid in the eluent is 10% to 30% by volume.
13. The process of claim 1, wherein in step (4), the concentration of glacial acetic acid in the eluent is 20% by volume.
14. The method of claim 1, wherein in step (4), the methanol concentration in the aqueous methanol solution is 40 to 80% by volume.
15. The method of claim 1, wherein in step (4), the methanol in the aqueous methanol solution has a concentration of 60% by volume.
16. The method according to any one of claims 1 to 15, wherein the extract to be extracted is a plant extract, preferably a peanut shell extract.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711115071.6A CN107880008B (en) | 2017-11-13 | 2017-11-13 | Method for extracting luteolin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711115071.6A CN107880008B (en) | 2017-11-13 | 2017-11-13 | Method for extracting luteolin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107880008A CN107880008A (en) | 2018-04-06 |
| CN107880008B true CN107880008B (en) | 2020-04-10 |
Family
ID=61780381
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711115071.6A Active CN107880008B (en) | 2017-11-13 | 2017-11-13 | Method for extracting luteolin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107880008B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114605368B (en) * | 2022-03-27 | 2023-05-26 | 中国药科大学 | Method for preparing luteolin ultrafine particles by using supercritical fluid enhanced solution dispersion technology |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1687054A (en) * | 2005-03-23 | 2005-10-26 | 浙江大学 | Method for preparing compound of luteolin |
-
2017
- 2017-11-13 CN CN201711115071.6A patent/CN107880008B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1687054A (en) * | 2005-03-23 | 2005-10-26 | 浙江大学 | Method for preparing compound of luteolin |
Non-Patent Citations (1)
| Title |
|---|
| Fe3O4磁性纳米粒子在中药成分分离分析中的应用研究;黎人恺;《CNKI硕士论文库》;20151001;全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107880008A (en) | 2018-04-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Kong et al. | Synthesis and characterization of the core–shell magnetic molecularly imprinted polymers (Fe3O4@ MIPs) adsorbents for effective extraction and determination of sulfonamides in the poultry feed | |
| Zhu et al. | A novel magnetically separable γ-Fe2O3/crosslinked chitosan adsorbent: preparation, characterization and adsorption application for removal of hazardous azo dye | |
| Zhang et al. | Preparation of carbon coated Fe3O4 nanoparticles and their application for solid-phase extraction of polycyclic aromatic hydrocarbons from environmental water samples | |
| Arica et al. | Polyaniline coated magnetic carboxymethylcellulose beads for selective removal of uranium ions from aqueous solution | |
| Cao et al. | Facile synthesis of a Ni (ii)-immobilized core–shell magnetic nanocomposite as an efficient affinity adsorbent for the depletion of abundant proteins from bovine blood | |
| Zhao et al. | Preparation of molecularly imprinted polymers based on magnetic carbon nanotubes for determination of sulfamethoxazole in food samples | |
| Wang et al. | Core–shell superparamagnetic Fe3O4@ β-CD composites for host–guest adsorption of polychlorinated biphenyls (PCBs) | |
| CN103285817B (en) | Amino acid modified silicon-structure-containing ferriferrous oxide nanoparticle and its application in dye adsorption treatment | |
| Duo et al. | Synthesis of magnetic metal–organic framework composites, Fe 3 O 4-NH 2@ MOF-235, for the magnetic solid-phase extraction of benzoylurea insecticides from honey, fruit juice and tap water samples | |
| CN109666110B (en) | Preparation method and application of tetracycline magnetic molecularly imprinted nanoparticles | |
| Weng et al. | Electrospun graphene oxide–doped nanofiber-based solid phase extraction followed by high-performance liquid chromatography for the determination of tetracycline antibiotic residues in food samples | |
| Zhou et al. | Magnetic nanoparticles speed up mechanochemical solid phase extraction with enhanced enrichment capability for organochlorines in plants | |
| CN103314021B (en) | Modified poly (styrene-co-maleic anhydride) and uses thereof | |
| Galhoum et al. | Chemical modifications of chitosan nano-based magnetic particles for enhanced uranyl sorption | |
| Hao et al. | Selective extraction of gallic acid in pomegranate rind using surface imprinting polymers over magnetic carbon nanotubes | |
| CN105597686B (en) | Fe3O4@MIL-100（Fe）Preparation method and applications | |
| Nicola et al. | Mesoporous magnetic nanocomposites: a promising adsorbent for the removal of dyes from aqueous solutions | |
| Wei et al. | Adsorption of bilirubin to magnetic multi-walled carbon nanotubes as a potential application in bound solute dialysis | |
| Zhang et al. | Synthesis of novel magnetic NiFe 2 O 4 nanocomposite grafted chitosan and the adsorption mechanism of Cr (VI) | |
| CN111871400B (en) | Preparation method and application of guanidine salt ionic liquid modified magnetic solid phase extraction adsorbent | |
| Yin et al. | Preparation of amino-functionalized covalent organic framework modified Fe3O4 nanoparticles for the selective enrichment of flavonoid glycosides | |
| Xiong et al. | Self‐assembly of alginate/polyethyleneimine multilayer onto magnetic microspheres as an effective adsorbent for removal of anionic dyes | |
| Yang et al. | Dual‐template magnetic molecularly imprinted particles with multi‐hollow structure for the detection of dicofol and chlorpyrifos‐methyl | |
| Wen et al. | Magnetic porous cellulose surface-imprinted polymers synthetized with assistance of deep eutectic solvent for specific recognition and purification of bisphenols | |
| Zhou et al. | Magnetic amino-functionalized metal-organic frameworks as a novel solid support in ionic liquids-based effervescent tablets for efficient extraction of polycyclic aromatic hydrocarbons in milks |
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 |