CN113975308A - Method for extracting total polyphenol from folium cortex eucommiae by using surfactant and microwave - Google Patents

Method for extracting total polyphenol from folium cortex eucommiae by using surfactant and microwave Download PDF

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CN113975308A
CN113975308A CN202111222886.0A CN202111222886A CN113975308A CN 113975308 A CN113975308 A CN 113975308A CN 202111222886 A CN202111222886 A CN 202111222886A CN 113975308 A CN113975308 A CN 113975308A
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彭密军
张光耀
王雪松
杨秋玲
黄韬
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Institute Of Testing And Analysis Guangdong Academy Of Sciences Guangzhou Analysis And Testing Center China
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Abstract

The invention discloses a method for extracting total polyphenol from folium cortex eucommiae by using a surfactant and microwave, which comprises the following steps: placing folium Eucommiae powder in water solution of surfactant, extracting under the assistance of microwave, performing solid-liquid separation to obtain extractive solution containing folium Eucommiae total polyphenols, further concentrating the extractive solution, and drying to obtain total polyphenols extract. The invention uses the water solution of the surfactant as the extracting solution, and combines the microwave extraction to extract the total polyphenol in the eucommia leaves, thereby greatly improving the extraction rate, reducing the extraction cost and avoiding the problem of toxic organic solvent residue.

Description

Method for extracting total polyphenol from folium cortex eucommiae by using surfactant and microwave
Technical Field
The invention relates to the technical field of active substance extraction methods, in particular to a method for extracting total polyphenol from eucommia leaves by using a surfactant and microwave.
Background
In recent 20 years, eucommia ulmoides leaves have become popular functional health food and botanical drug materials in the middle and two countries. The folium Eucommiae extract has effects of reducing cholesterol, lowering blood pressure, and preventing fatty liver. The folium Eucommiae is rich in bioactive components such as polyphenolic acid (such as chlorogenic acid), flavonoid (such as quercetin and rutin), iridoid (such as aucubin and geniposide), and nutrients such as amino acids, vitamins and minerals.
At present, the traditional eucommia ulmoides leaf polyphenol extraction methods comprise a water decoction method, an alcohol extraction method, an enzymolysis method and the like. However, the traditional water decoction method has low extraction efficiency, and the eucommia leaves are easily inactivated in a high-temperature environment for a long time; the alcohol extraction method has high consumption of organic solvent, needs to be recycled, has organic solvent residue in the extract and has serious environmental pollution; the enzymolysis method is easily influenced by various factors such as the type, the dosage, the enzymolysis time, the temperature and the like of enzyme, and the production link is difficult to control. Therefore, the development of the method for extracting the eucommia ulmoides leaf polyphenol, which has the advantages of low energy consumption, short time, clean process, simple and efficient process and easy industrial production, is very important.
Disclosure of Invention
The invention aims to provide a method for extracting total polyphenol from folium cortex eucommiae by using a surfactant and microwave in cooperation, aiming at the defects and shortcomings of the prior art.
In order to achieve the purpose, the technical scheme provided by the invention is as follows:
a method for extracting total polyphenols from folium Eucommiae with surfactant and microwave comprises: placing folium Eucommiae powder in water solution of surfactant, extracting under the assistance of microwave, performing solid-liquid separation to obtain extractive solution containing folium Eucommiae total polyphenols, further concentrating the extractive solution, and drying to obtain total polyphenols extract.
As a further improvement of the above scheme, the method for extracting total polyphenols from folium cortex eucommiae by using surfactant and microwave comprises the following steps:
(1) drying folium Eucommiae, pulverizing, and sieving to obtain folium Eucommiae powder;
(2) placing eucommia leaf powder into an aqueous solution containing a surfactant according to a material-liquid ratio of 1: 1-50 g/mL, uniformly mixing, and standing for 10-100 min to obtain a mixed solution;
(3) performing microwave extraction on the mixed solution obtained in the step (2), wherein the power of the microwave is 200-300W, and the extraction time is 1-3 min;
(4) after the microwave extraction is finished, filtering the mixed solution to obtain filtrate;
(5) and (3) repeating the steps (1) to (4) on filter residues, finally combining the filtrate with the filtrate obtained in the step (4) to obtain an extracting solution containing the total polyphenol of the eucommia ulmoides leaves, and further concentrating and drying the extracting solution to obtain the total polyphenol extract.
As a further improvement of the scheme, in the step (2), the surfactant accounts for 0.5-3% of the aqueous solution by mass percent.
As a further improvement of the scheme, in the step (1), the drying temperature is 50-70 ℃, preferably 50 ℃.
As a further improvement of the above aspect, the surfactant comprises one or more of sodium lauroyl glutamate, cocamidopropyl betaine, polyglycerol laurate.
As a further improvement of the above scheme, in the step (5), the drying manner is selected from any one of reduced pressure drying, spray drying, vacuum drying and microwave drying.
As a further improvement of the scheme, in the step (1), the mesh number of the sieve is 40-100 meshes, and preferably 60 meshes.
The principle of the invention is as follows: the surfactant has a specific amphiphilic molecular structure, can effectively reduce the interfacial tension between the eucommia ulmoides leaf cell membrane and an aqueous solution, enhances the wettability and permeability of a solvent to materials, and has a solubilizing effect on total polyphenols in the eucommia ulmoides leaves. In addition, the microwave radiation process has penetrability, and the temperature in the cell rises rapidly due to the absorption of microwave energy, so that the pressure in the cell exceeds the expansion bearing capacity of the cell wall, the cell is broken, and the active ingredients in the cell flow out.
Compared with the prior art, the invention has the beneficial effects that:
1. the total polyphenol of the eucommia leaves is extracted by adopting a method of combining a surfactant and microwaves, the extraction time is shortened from the traditional 2-5 hours to 40-60 min, the extraction time is greatly shortened, the energy consumption and the cost in the production process are greatly reduced, and the extraction efficiency is improved.
2. Distilled water solution dissolved with a small amount of surfactant is used as extracting solution to replace the traditional organic solvent, and the problem of organic solvent residue is avoided because toxic organic solvent is not contained.
3. The total polyphenol extracted from folium cortex eucommiae by the method avoids long-time heat effect, and reduces the degree of thermal decomposition and oxidation of the polyphenol.
4. The method of the invention has no high requirement on the external environment, and simultaneously, the required equipment and the operation method are simple and the investment cost is less.
Drawings
Figure 1 is a standard graph of total polyphenol concentration.
Detailed Description
To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to specific examples.
The method for determining the yield of the total polyphenol in the eucommia ulmoides leaves (Folin-ciocalteu (FC) method) is as follows:
determination of the standard curve: weighing 2.060mg of gallic acid (GA, relative molecular mass 188.14), adding 70% methanol into a 10mL volumetric flask to dissolve and fix the volume to the scale, preparing mother liquor, and shaking up (in-situ). The concentrations of the mother liquors were 20.6. mu.g/mL, 41.2. mu.g/mL, 61.8. mu.g/mL, 82.4. mu.g/mL and 103. mu.g/mL, respectively.
As can be seen from the total polyphenol concentration standard curve in fig. 1, the regression equation and the correlation coefficient of the absorbance value and the standard solution concentration are respectively 0.0085x +0.09(R ═ 0.0085x + 0.09)20.9993), indicating good linearity over this concentration range.
Determination of polyphenol content in samples: using a pipette to respectively transfer 1.0mL of sample liquid into graduated test tubes, respectively adding 5.0mL of 10% forrin phenol reagent into each test tube, and shaking up. Reacting for 3-8 min, adding 4.0mL of 7.5% sodium carbonate (Na)2CO3) Adding water to the solution, fixing the volume to the scale, and shaking up. Standing at room temperature for 60 min. The absorbance was measured with a spectrophotometer at a wavelength of 765nm in a 10mL cuvette.
Example 1:
a method for extracting total polyphenol from folium Eucommiae by using surfactant and microwave comprises the following steps:
(1) drying folium Eucommiae, pulverizing, and sieving with 60 mesh sieve to obtain folium Eucommiae powder;
(2) accurately weighing 1g of folium Eucommiae powder, adding into 1% sodium lauroyl glutamate-containing water solution at a material-to-liquid ratio of 1:26g/mL, mixing, and standing for 1h to obtain mixed solution;
(3) performing microwave extraction on the mixed solution in the step (2), wherein the microwave power is 200W, and the extraction time is 2.5 min;
(4) after the microwave extraction is finished, filtering the mixed solution to obtain filtrate;
(5) and (3) repeating the steps (1) to (4) on filter residues, finally combining the filtrate with the filtrate obtained in the step (4) to obtain an extracting solution containing the total polyphenol of the eucommia ulmoides leaves, and further concentrating and drying the extracting solution to obtain the total polyphenol extract.
The total polyphenol content of the extract obtained above was measured by the Folin-ciocalteu (FC) method, and the results showed that: the yield of total polyphenols in the extract was 6.02%.
Example 2:
a method for extracting total polyphenol from folium Eucommiae by using surfactant and microwave comprises the following steps:
(1) drying folium Eucommiae, pulverizing, and sieving with 60 mesh sieve to obtain folium Eucommiae powder;
(2) accurately weighing 1g of folium Eucommiae powder, adding into an aqueous solution containing 2% sodium lauroyl glutamate at a material-to-liquid ratio of 1:26g/mL, mixing well, and standing for 1h to obtain a mixed solution;
(3) performing microwave extraction on the mixed solution in the step (2), wherein the microwave power is 200W, and the extraction time is 2.5 min;
(4) after the microwave extraction is finished, filtering the mixed solution to obtain filtrate;
(5) and (3) repeating the steps (1) to (4) on filter residues, finally combining the filtrate with the filtrate obtained in the step (4) to obtain an extracting solution containing the total polyphenol of the eucommia ulmoides leaves, and further concentrating and drying the extracting solution to obtain the total polyphenol extract.
The total polyphenol content of the extract obtained above was measured by the Folin-ciocalteu (FC) method, and the results showed that: the yield of total polyphenols in the extract was 6.12%.
Example 3:
a method for extracting total polyphenol from folium Eucommiae by using surfactant and microwave comprises the following steps:
(1) drying folium Eucommiae, pulverizing, and sieving with 60 mesh sieve to obtain folium Eucommiae powder;
(2) accurately weighing 1g of folium Eucommiae powder, adding into 3% sodium lauroyl glutamate-containing water solution at a material-to-liquid ratio of 1:26g/mL, mixing, and standing for 1h to obtain mixed solution;
(3) performing microwave extraction on the mixed solution in the step (2), wherein the microwave power is 200W, and the extraction time is 2.5 min;
(4) after the microwave extraction is finished, filtering the mixed solution to obtain filtrate;
(5) and (3) repeating the steps (1) to (4) on filter residues, finally combining the filtrate with the filtrate obtained in the step (4) to obtain an extracting solution containing the total polyphenol of the eucommia ulmoides leaves, and further concentrating and drying the extracting solution to obtain the total polyphenol extract.
The total polyphenol content of the extract obtained above was measured by the Folin-ciocalteu (FC) method, and the results showed that: the yield of total polyphenols in the extract was 6.08%.
Example 4:
a method for extracting total polyphenol from folium Eucommiae by using surfactant and microwave comprises the following steps:
(1) drying folium Eucommiae, pulverizing, and sieving with 60 mesh sieve to obtain folium Eucommiae powder;
(2) accurately weighing 1g of folium Eucommiae powder, adding into 1% sodium lauroyl glutamate-containing water solution at a material-to-liquid ratio of 1:20g/mL, mixing, and standing for 1h to obtain mixed solution;
(3) performing microwave extraction on the mixed solution in the step (2), wherein the microwave power is 200W, and the extraction time is 2.5 min;
(4) after the microwave extraction is finished, filtering the mixed solution to obtain filtrate;
(5) and (3) repeating the steps (1) to (4) on filter residues, finally combining the filtrate with the filtrate obtained in the step (4) to obtain an extracting solution containing the total polyphenol of the eucommia ulmoides leaves, and further concentrating and drying the extracting solution to obtain the total polyphenol extract.
The total polyphenol content of the extract obtained above was measured by the Folin-ciocalteu (FC) method, and the results showed that: the yield of total polyphenols in the extract was 5.75%.
Example 5:
a method for extracting total polyphenol from folium Eucommiae by using surfactant and microwave comprises the following steps:
(1) drying folium Eucommiae, pulverizing, and sieving with 60 mesh sieve to obtain folium Eucommiae powder;
(2) accurately weighing 1g of folium cortex eucommiae powder, adding the folium cortex eucommiae powder into an aqueous solution containing 1% of decaglycerol laurate according to the material-liquid ratio of 1:26g/mL, uniformly mixing, and standing for 1h to obtain a mixed solution;
(3) performing microwave extraction on the mixed solution in the step (2), wherein the microwave power is 200W, and the extraction time is 2.5 min;
(4) after the microwave extraction is finished, filtering the mixed solution to obtain filtrate;
(5) and (3) repeating the steps (1) to (4) on filter residues, finally combining the filtrate with the filtrate obtained in the step (4) to obtain an extracting solution containing the total polyphenol of the eucommia ulmoides leaves, and further concentrating and drying the extracting solution to obtain the total polyphenol extract.
The total polyphenol content of the extract obtained above was measured by the Folin-ciocalteu (FC) method, and the results showed that: the yield of total polyphenols in the extract was 5.88%.
Example 6:
a method for extracting total polyphenol from folium Eucommiae by using surfactant and microwave comprises the following steps:
(1) drying folium Eucommiae, pulverizing, and sieving with 40 mesh sieve to obtain folium Eucommiae powder;
(2) accurately weighing 1g of folium cortex eucommiae powder, adding the folium cortex eucommiae powder into an aqueous solution containing 1% of cocamidopropyl betaine according to the material-liquid ratio of 1:26g/mL, uniformly mixing, and standing for 1h to obtain a mixed solution;
(3) performing microwave extraction on the mixed solution in the step (2), wherein the microwave power is 200W, and the extraction time is 2.5 min;
(4) after the microwave extraction is finished, filtering the mixed solution to obtain filtrate;
(5) and (3) repeating the steps (1) to (4) on filter residues, finally combining the filtrate with the filtrate obtained in the step (4) to obtain an extracting solution containing the total polyphenol of the eucommia ulmoides leaves, and further concentrating and drying the extracting solution to obtain the total polyphenol extract.
The total polyphenol content of the extract obtained above was measured by the Folin-ciocalteu (FC) method, and the results showed that: the yield of total polyphenols in the extract was 5.65%.
Example 7:
a method for extracting total polyphenol from folium Eucommiae by using surfactant and microwave comprises the following steps:
(1) drying folium Eucommiae, pulverizing, and sieving with 80 mesh sieve to obtain folium Eucommiae powder;
(2) accurately weighing 1g of folium Eucommiae powder, adding into an aqueous solution containing 0.5% of cocamidopropyl betaine at a material-to-liquid ratio of 1:40g/mL, mixing, and standing for 10min to obtain a mixed solution;
(3) performing microwave extraction on the mixed solution in the step (2), wherein the microwave power is 200W, and the extraction time is 1 min;
(4) after the microwave extraction is finished, filtering the mixed solution to obtain filtrate;
(5) and (3) repeating the steps (1) to (4) on filter residues, finally combining the filtrate with the filtrate obtained in the step (4) to obtain an extracting solution containing the total polyphenol of the eucommia ulmoides leaves, and further concentrating and drying the extracting solution to obtain the total polyphenol extract.
The total polyphenol content of the extract obtained above was measured by the Folin-ciocalteu (FC) method, and the results showed that: the yield of total polyphenols in the extract was 5.35%.
Example 8:
a method for extracting total polyphenol from folium Eucommiae by using surfactant and microwave comprises the following steps:
(1) drying folium Eucommiae, pulverizing, and sieving with 100 mesh sieve to obtain folium Eucommiae powder;
(2) accurately weighing 1g of folium Eucommiae powder, adding into 1.5% aqueous solution of cocamidopropyl betaine at a material-to-liquid ratio of 1:50g/mL, mixing, and standing for 100min to obtain mixed solution;
(3) performing microwave extraction on the mixed solution in the step (2), wherein the microwave power is 300W, and the extraction time is 3 min;
(4) after the microwave extraction is finished, filtering the mixed solution to obtain filtrate;
(5) and (3) repeating the steps (1) to (4) on filter residues, finally combining the filtrate with the filtrate obtained in the step (4) to obtain an extracting solution containing the total polyphenol of the eucommia ulmoides leaves, and further concentrating and drying the extracting solution to obtain the total polyphenol extract.
The total polyphenol content of the extract obtained above was measured by the Folin-ciocalteu (FC) method, and the results showed that: the yield of total polyphenols in the extract was 5.52%.
Comparative example 1:
a method for extracting total polyphenol from folium Eucommiae specifically comprises the following steps:
(1) drying folium Eucommiae, pulverizing, and sieving with 60 mesh sieve to obtain folium Eucommiae powder;
(2) accurately weighing 1g of folium cortex eucommiae powder, adding distilled water according to the material-liquid ratio of 1:26g/mL, uniformly mixing, and standing for 1h to obtain a mixed solution;
(3) performing microwave extraction on the mixed solution in the step (2), wherein the microwave power is 200W, and the extraction time is 2.5 min;
(4) after the microwave extraction is finished, filtering the mixed solution to obtain filtrate;
(5) and (3) repeating the steps (1) to (4) on filter residues, finally combining the filtrate with the filtrate obtained in the step (4) to obtain an extracting solution containing the total polyphenol of the eucommia ulmoides leaves, and further concentrating and drying the extracting solution to obtain the total polyphenol extract.
The total polyphenol content of the extract obtained above was measured by the Folin-ciocalteu (FC) method, and the results showed that: the yield of total polyphenols in the extract was 4.58%.
Comparative example 2:
a method for extracting total polyphenol from folium Eucommiae specifically comprises the following steps:
(1) drying folium Eucommiae, pulverizing, and sieving with 60 mesh sieve to obtain folium Eucommiae powder;
(2) accurately weighing 1g of folium cortex eucommiae powder, adding distilled water according to the material-liquid ratio of 1:26g/mL, uniformly mixing, and standing for 1h to obtain a mixed solution;
(3) filtering the mixed solution to obtain filtrate;
(4) and (3) repeating the steps (1) to (3) on filter residues, finally combining the filtrate with the filtrate obtained in the step (3) to obtain an extracting solution containing the total polyphenol of the eucommia ulmoides leaves, and further concentrating and drying the extracting solution to obtain the total polyphenol extract.
The total polyphenol content of the extract obtained above was measured by the Folin-ciocalteu (FC) method, and the results showed that: the yield of total polyphenols in the extract was 2.43%.
Comparative examples 1, 2 and example 2 were compared, and the results are shown in table 1 below.
TABLE 1 comparison of test results of total polyphenols extracted from folium Eucommiae by different methods
Figure BDA0003313277900000091
The test results of extracting total polyphenol from folium cortex eucommiae by comparing different methods in table 1 can show that: the extraction method of the invention can obviously improve the yield of total polyphenol extracted from the eucommia leaves by combining the water solution of the surfactant with microwave extraction.
The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and substitutions can be made without departing from the technical principle of the present invention, and these modifications and substitutions should also be regarded as the protection scope of the present invention.

Claims (9)

1. A method for extracting total polyphenol from eucommia ulmoides leaves by using a surfactant and microwave, which is characterized by comprising the following steps: placing folium Eucommiae powder in water solution of surfactant, extracting under the assistance of microwave, performing solid-liquid separation to obtain extractive solution containing folium Eucommiae total polyphenols, further concentrating the extractive solution, and drying to obtain total polyphenols extract.
2. The method for extracting total polyphenols from folium Eucommiae by using surfactant and microwave as claimed in claim 1, which comprises the following steps:
(1) drying folium Eucommiae, pulverizing, and sieving to obtain folium Eucommiae powder;
(2) placing eucommia leaf powder into an aqueous solution containing a surfactant according to a material-liquid ratio of 1: 1-50 g/mL, uniformly mixing, and standing for 10-100 min to obtain a mixed solution;
(3) performing microwave extraction on the mixed solution obtained in the step (2), wherein the power of the microwave is 200-300W, and the extraction time is 1-3 min;
(4) after the microwave extraction is finished, filtering the mixed solution to obtain filtrate;
(5) and (3) repeating the steps (1) to (4) on filter residues, finally combining the filtrate with the filtrate obtained in the step (4) to obtain an extracting solution containing the total polyphenol of the eucommia ulmoides leaves, and further concentrating and drying the extracting solution to obtain the total polyphenol extract.
3. The method for extracting total polyphenols from folium cortex eucommiae by using the surfactant and microwave in cooperation with claim 2, wherein in the step (2), the surfactant accounts for 0.5-3% of the aqueous solution by mass.
4. The method for extracting total polyphenols from folium Eucommiae using surfactant in combination with microwave as claimed in claim 2, wherein the drying temperature in step (1) is 50-70 ℃.
5. The method for extracting total polyphenols from folium Eucommiae using surfactant in combination with microwave as claimed in claim 4, wherein the drying temperature in step (1) is 50 ℃.
6. The method for extracting total polyphenol from eucommia ulmoides leaves by using the surfactant in cooperation with microwave according to claim 1 or 2, wherein the surfactant comprises one or more of sodium lauroyl glutamate, cocamidopropyl betaine and polyglycerol laurate.
7. The method for extracting total polyphenols from folium Eucommiae using surfactant in combination with microwave as claimed in claim 1, wherein in the step (5), the drying manner is selected from one of reduced pressure drying, spray drying, vacuum drying and microwave drying.
8. The method for extracting total polyphenols from folium Eucommiae using surfactant and microwave as claimed in claim 1, wherein in step (1), the mesh number of the sieve is 40-100 meshes.
9. The method for extracting total polyphenols from folium Eucommiae using surfactant in combination with microwave as claimed in claim 1, wherein in step (1), the mesh number of the sieve is 60 meshes.
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