CN115400165A - Eucalyptus leaf extract, preparation method and application thereof in preparation of weight-losing and lipid-lowering products - Google Patents
Eucalyptus leaf extract, preparation method and application thereof in preparation of weight-losing and lipid-lowering products Download PDFInfo
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- CN115400165A CN115400165A CN202211003304.4A CN202211003304A CN115400165A CN 115400165 A CN115400165 A CN 115400165A CN 202211003304 A CN202211003304 A CN 202211003304A CN 115400165 A CN115400165 A CN 115400165A
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- China
- Prior art keywords
- eucalyptus
- eucalyptus leaf
- leaf extract
- preparation
- extraction
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Links
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- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention provides a eucalyptus leaf extract, a preparation method and application thereof in preparation of weight-losing and lipid-lowering products, and belongs to the technical field of plant extraction. The invention solves the problems of low extraction rate of effective components of eucalyptus leaves, poor activity of extracted products and the like in the prior art by adopting multi-solvent low-temperature continuous phase change extraction, column chromatography separation and other processes. The preparation method of the eucalyptus leaf extract can realize higher impurity removal rate, and has simple operation steps and high yield. The eucalyptus leaf extract obtained by the preparation method has higher polyphenol content and higher total yield, has good weight-losing and lipid-lowering effects, can effectively control the body weight, regulate the blood lipid level, and effectively prevent and improve obesity. In addition, the eucalyptus leaf resource is rich and low in price, the preparation method is simple, the cost is low, the requirement of industrial production is met, and the eucalyptus leaf tea has practical significance and application value for developing medicines or health care products, so that the utilization rate of the eucalyptus leaf resource is improved.
Description
Technical Field
The invention relates to the technical field of plant extraction, and particularly relates to a eucalyptus leaf extract, a preparation method and application thereof in preparation of weight-losing and lipid-lowering products.
Background
In recent years, with the improvement of living standard, the incidence of nutritional obesity caused by unhealthy work and rest and eating modes is increasing. Nutritional obesity has been reported to increase the risk of hypertension, type ii diabetes, coronary heart disease and cancer, greatly affecting the health of the body. Therefore, how to effectively prevent and treat obesity has become one of public safety and health problems worldwide. Although weight loss drugs are effective in controlling body weight, long-term administration may have adverse effects on the body. Therefore, the market demand for developing safe, non-toxic and side-effect natural products with the functions of losing weight and reducing blood fat is gradually released, and the future industry development prospect is good.
China has rich eucalyptus resources, and has the characteristics of large area, wide region, multiple varieties and the like. The eucalyptus leaf serving as a byproduct of the eucalyptus is always used as a folk traditional Chinese herbal medicine and is usually used for clearing heat, detoxifying, treating cold and the like. Studies show that the eucalyptus leaf extract contains a large amount of polyphenols, shows strong antioxidant activity in vivo and in vitro, and has a series of effects of resisting aging, tumors, diabetes, inflammation and the like which are reported, but the existing methods such as a chemical method and an enzymatic method have low extraction rate of active ingredients of the eucalyptus leaf and poor activity of extracted products, and limit the application of the eucalyptus leaf extract in industry. At present, no reports related to prevention and treatment of obesity by using eucalyptus leaf extract are seen.
Disclosure of Invention
The invention aims to provide a eucalyptus leaf extract, a preparation method and application thereof in preparing weight-losing and lipid-lowering products, and solves the problems of low extraction rate of effective ingredients of eucalyptus leaves, poor activity of extracted products and the like in the prior art.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of a eucalyptus leaf extract, which comprises the following steps:
pulverizing folium Eucalypti Globueli to obtain folium Eucalypti Globueli powder;
performing continuous phase change extraction on the eucalyptus leaf powder to obtain an extracting solution;
sequentially concentrating and spray drying the extracting solution to obtain a crude extract of the eucalyptus leaves;
purifying the eucalyptus crude extract to obtain a eucalyptus extract;
the extractant adopted by the continuous phase-change extraction is butane, ethanol aqueous solution and water in sequence.
Preferably, the particle size of the eucalyptus leaf powder is less than 550 μm.
Preferably, the extraction pressure of the continuous phase-change extraction is 0.5-1.5 MPa, the extraction temperature is 55-65 ℃, the resolution temperature is 65-75 ℃, and the extraction time of each extractant is 80-120 min.
Preferably, the purification is performed by silica gel column separation, and the mesh number of the silica gel column used for silica gel column separation is 100-300 meshes.
Preferably, gradient elution is adopted in the silica gel column separation process, and the elution flow rate of the gradient elution is 30-50 ml/min.
Preferably, the eluent adopted by the silica gel column separation comprises a component A and a component B;
the component A is ethanol, and the component B is ethyl acetate or petroleum ether.
Preferably, during the gradient elution, the volume ratio of component a to component B is in the order of 0;
eluting 2-4 column volumes of eluent with each volume ratio.
Preferably, the eluted product is concentrated and spray-dried to obtain the eucalyptus leaf extract.
The invention also provides a eucalyptus leaf extract prepared by the preparation method, which comprises 10-20% of oenothein B, 1-2% of terlimacin I, 1-1.5% of quercetin-3-O-glucuronide, 0.8-1.2% of gallic acid, 0.2-0.5% of terlimacin II, 3-6% of 1,2,3,4,6 pentagalloyl glucose, and the content of total phenols in the eucalyptus leaf extract is 50-60%.
The invention also provides application of the eucalyptus leaf extract in preparing a product for losing weight and reducing fat.
The invention has the technical effects and advantages that:
the preparation method of the eucalyptus leaf extract can realize higher impurity removal rate, and has simple operation steps and high yield. The eucalyptus leaf extract obtained by the preparation method has higher polyphenol content and higher total yield, and is suitable for application in actual industry. The detection proves that the eucalyptus leaf extract obtained by the method has good weight-losing and lipid-lowering effects, can effectively control the body weight, regulate the blood lipid level, and effectively prevent and improve obesity. In addition, the eucalyptus leaf resource is rich and low in price, the preparation method is simple, the cost is low, the requirements of industrial production are met, the eucalyptus leaf tea has practical significance and application value for developing medicines or health care products, and the utilization rate of the eucalyptus leaf resource is improved to a certain extent.
Drawings
FIG. 1 shows the results of liquid chromatography;
FIG. 2 shows the measurement results of total triglyceride content in mouse serum;
FIG. 3 shows the measurement of total cholesterol content in mouse serum;
FIG. 4 shows the measurement results of low density lipoprotein content in mouse serum;
FIG. 5 shows the measurement results of high density lipoprotein content in mouse serum.
Detailed Description
The invention provides a preparation method of a eucalyptus leaf extract, which comprises the following steps:
pulverizing folium Eucalypti Globueli to obtain folium Eucalypti Globueli powder;
performing continuous phase change extraction on the eucalyptus leaf powder to obtain an extracting solution;
sequentially concentrating and spray drying the extracting solution to obtain a crude extract of the eucalyptus leaves;
purifying the eucalyptus crude extract to obtain a eucalyptus extract;
the extraction agents adopted by the continuous phase-change extraction are butane, ethanol aqueous solution and water.
In the present invention, the eucalyptus leaves are preferably dried in the shade and then pulverized to obtain eucalyptus leaf powder, and the particle size of the eucalyptus leaf powder is preferably 550 μm or less, and more preferably 350 μm or less; performing continuous phase-change extraction on the eucalyptus powder, wherein the continuous phase-change extraction is preferably performed by using a continuous phase-change extraction kettle, the extraction pressure of the continuous phase-change extraction is preferably 0.5-1.5 MPa, the further preference is 0.8-1.2 MPa, the extraction temperature is preferably 55-65 ℃, the further preference is 58-62 ℃, the analysis temperature is preferably 65-75 ℃, the further preference is 68-72 ℃, the extraction time of each time is preferably 80-120 min, and the further preference is 90-110 min; the extraction agents adopted by the continuous phase-change extraction are butane, ethanol aqueous solution and water, the invention has no requirement on the extraction sequence, and the volume concentration of the ethanol aqueous solution is preferably 60-80%.
In the invention, the concentration is preferably performed by vacuum reduced pressure concentration, the temperature of the vacuum reduced pressure concentration is preferably 55-70 ℃, the vacuum degree is preferably 300-500 Pa, the air inlet temperature of the spray drying is preferably 200-250 ℃, the feeding speed is preferably 80-120 rpm, and the frequency of an atomizer is preferably 40-50 Hz.
In the present invention, the purification is preferably performed by silica gel column separation, and the mesh number of the silica gel column used for silica gel column separation is preferably 100 to 300 meshes, and more preferably 150 to 250 meshes; gradient elution is preferably adopted in the silica gel column separation process, and the elution flow rate of the gradient elution is preferably 30-50 ml/min, and further preferably 35-45 ml/min; the eluent adopted by the silica gel column separation preferably comprises a component A and a component B, wherein the component A is preferably ethanol, and the component B is preferably ethyl acetate or petroleum ether; during the gradient elution, the volume ratio of component a to component B is preferably in the order of 0; in the invention, 2-4 column volumes of eluent in each volume ratio are preferably eluted, after gradient elution is finished, the eluted product is preferably concentrated and spray-dried to obtain the eucalyptus leaf extract, the concentration is preferably vacuum concentration, the temperature of the vacuum concentration is preferably 65-85 ℃, the vacuum degree is preferably 400-600 Pa, the air inlet temperature of the spray-drying is preferably 250-300 ℃, the feeding speed is preferably 100-150 rpm, and the frequency of an atomizer is preferably 50-60 Hz.
The invention also provides the eucalyptus leaf extract prepared by the preparation method.
The invention also provides application of the eucalyptus leaf extract in preparing a weight-losing and lipid-lowering product, wherein the weight-losing and lipid-lowering product is preferably a health-care product or a medicine; the product preferably takes the eucalyptus leaf extract as the only active component; the product preferably also comprises auxiliary materials, the type of the auxiliary materials is not specially limited, and the auxiliary materials are conventional in the field; the product dosage form of the invention is preferably powder, tablets, granules, capsules, solutions, emulsions, suspensions, injections, sprays, aerosols or powder mists and the like. In the present invention, the product comprises a carrier, which is preferably a binder, a lubricant, a disintegrant, a cosolvent, a diluent, a stabilizer, a suspending agent, a pigment, a flavoring agent, etc. for oral preparation; preservatives, solubilizers, stabilizers and the like for injectable preparations; bases for topical formulations, diluents, lubricants, preservatives, and the like.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
The method comprises the steps of drying eucalyptus leaves of Guanglin No. 9 in the shade, mechanically crushing, crushing to 30 meshes, weighing 20kg of eucalyptus leaf powder, putting the powder into a continuous phase-change extraction kettle, sealing and extracting, wherein the extraction pressure is 1.0MPa, the extraction temperature is 65 ℃, the resolution temperature is 75 ℃, butane, 70% ethanol and water are sequentially used as extracting agents, and the extraction time is 100min each time, so that the eucalyptus leaf extracting solution is obtained. Vacuum concentrating folium Eucalypti Globueli extractive solution at 60 deg.C under 400Pa, and spray drying at air inlet temperature of 200 deg.C, feeding speed of 100rpm, and atomizer frequency of 45Hz to obtain folium Eucalypti Globueli crude extract. The crude extract of the eucalyptus leaves passes through a 100-mesh silica gel column, the elution flow rate is 40ml/min, the gradient elution is carried out by using ethanol and ethyl acetate, the volume ratio of the ethanol to the ethyl acetate is 0.
Example 2
Drying Eucalyptus globulus leaf raw material 'Guanglin No. 9' in shade, mechanically pulverizing, pulverizing to 40 meshes, weighing Eucalyptus globulus leaf powder 15kg, placing into a continuous phase-change extraction kettle, sealing and extracting, wherein the extraction pressure is 1.2MPa, the extraction temperature is 60 ℃, the resolution temperature is 70 ℃, sequentially using butane, 65% ethanol and water as extracting agents, and extracting for 120min each time, and collecting the Eucalyptus globulus leaf extract. Concentrating folium Eucalypti Globueli extractive solution under vacuum at 65 deg.C and 500Pa, and spray drying at air inlet temperature of 250 deg.C, feeding speed of 150rpm, and atomizer frequency of 50Hz. Passing the crude extract of the eucalyptus leaves through a 200-mesh silica gel column, eluting at the flow rate of 35ml/min, performing gradient elution by using ethanol and ethyl acetate, wherein the volume ratio of the ethanol to the ethyl acetate is 0.
Example 3
Drying Eucalyptus globulus leaf raw material 'Guanglin No. 9' in shade, mechanically pulverizing, pulverizing to 60 meshes, weighing 15kg of Eucalyptus globulus leaf powder, placing into a continuous phase-change extraction kettle, sealing and extracting at 65 ℃ and 75 ℃ under the extraction pressure of 1.2MPa, respectively using butane, 80% ethanol and water as extracting agents for 120min each time, and collecting Eucalyptus globulus leaf extract. Vacuum concentrating folium Eucalypti Globueli extractive solution at 60 deg.C under 400Pa, and spray drying at air inlet temperature of 200 deg.C, feeding speed of 100rpm, and atomizer frequency of 45Hz to obtain folium Eucalypti Globueli crude extract. The crude extract of the eucalyptus leaves is put into a 200-mesh silica gel column, the flow rate of elution is 35ml/min, gradient elution is carried out by using ethanol and petroleum ether, the volume ratio of the ethanol to the petroleum ether is 0.
Experimental example 1 liquid chromatography analysis
The eucalyptus leaf extract obtained in example 1 was subjected to liquid chromatography under conditions of a Diamonsil C18 column (250X 4.6mm,5 μm); the detection wavelength is 270nm; the mobile phase is 10 to 90 percent methanol and is eluted for 1 hour in a gradient way, and the flow rate is 1mL/mim; the amount of the sample was 20. Mu.L.
The results are shown in FIG. 1, where: 1. gemin D; 2. evening primrose essence B; 3. 5-methoxyfurfural; 4. quercus castellata extract; 5. a teriomacin I; 6. chlorogenic acid; 7. (iii) terlimacin ii; 8. 1,2,3,6-tetragalloylglucose; 9. 4, gallic acid ethyl ester; 10. 1,2,3,4,6-pentagalloylglucose; 11. Quercetin-3-O-glucuronide.
Experimental example 2 determination of extraction yield and Total phenol content
The amount of the eucalyptus extract in examples 1 and 2 was measured, and the extraction rate was calculated (extraction rate = amount of the eucalyptus extract/amount of the eucalyptus powder × 100%);
the total phenol content was determined by the Folin's phenol method, and the results are shown in Table 1 below:
TABLE 1 extraction results of eucalyptus leaf extract
| Quality of eucalyptus leaf extract | Extraction rate | Total phenol content | |
| Example 1 | 11.2kg | 56％ | 55％ |
| Example 2 | 9.0kg | 60％ | 50％ |
EXAMPLE 3 study of weight-reducing and lipid-lowering actions
Test animals: male, SPF grade C57BL/6J mice, weighing approximately (23 + -2 g), purchased from Experimental animals technology, inc. of Viton, beijing;
test drugs and reagents: eucalyptus extract, having a total phenolic content of about 55%; physiological saline; and (3) reagent kit; kits for Total Cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL-C), high-density lipoprotein (HDL-C), etc.;
feed: 60% high fat model feed, 10% low fat control feed were purchased from souton telofil feed technologies ltd;
the experimental method comprises the following steps: after 7 days of acclimation, 32C 57BL/6J mice were divided into four groups, namely a normal control group, a model control group, a low dose group of eucalyptus extract (50 mg/kg) and a high dose group of eucalyptus extract (100 mg/kg). The normal control group was fed with 10% low-fat control diet, while the model group and the low and high dose eucalyptus leaf extract group were fed with 60% high-fat model diet for 10 weeks. Beginning at 1d, normal control group mice and model control group mice are subjected to intragastric administration of normal saline, and low-dose and high-dose eucalyptus leaf extract group mice are subjected to intragastric administration of 50mg/kg and 100mg/kg of eucalyptus leaf extracts respectively. After the feeding period is finished, the mice are sacrificed, and serum, organs and fat are collected and stored at-80 ℃ for testing.
The weight, food intake, organ coefficient, fat coefficient, serum lipid level, and the like of the mice were measured, and the results are as follows. All experimental data were analyzed using SPSS 17.0 statistical software. Single-factor analysis of variance (One-wayAnova) is adopted for comparison among multiple groups, and a Duncan method is adopted for pairwise comparison. The results are expressed as mean ± sd, and differences with p <0.05 are significant.
During the test, the body weights of the mice in each group were measured and recorded every week, and the body weights and the body weight gains of the mice in each group are shown in table 2.
TABLE 2 weight gain (g) of mice
Note: the data in the table are the mean ± standard deviation of the samples; p <0.01 compared to normal control group, # p <0.05 compared to model control group.
As can be seen from Table 2, the weight growth rate of the mice in the model control group is significantly higher than that of the normal control group, which indicates that the establishment of the obesity model of the mice is successful. Compared with a model control group, the weight gain of mice in a low and high dose group of the eucalyptus leaf extract is reduced, wherein the weight reducing effect of the high dose group is very obvious. The result shows that the eucalyptus leaf extract can effectively inhibit the body obesity caused by high-fat diet after a certain dose of the eucalyptus leaf extract is taken, and the weight-losing effect is good.
The test weighs epididymal fat, inguinal fat, perirenal fat, brown fat and liver of each group of mice, and calculates the index thereof, and the results are shown in table 3.
Table 3 mouse liver coefficient and fat coefficient (%)
Note: the data in the table are the mean ± standard deviation of the samples; p <0.01 compared to normal control group, # p <0.05 compared to model control group.
As can be seen from Table 3, the liver coefficients of mice in the low and high dose groups of the eucalyptus leaf extract are significantly lower than those of the model control group, which indicates that the eucalyptus leaf extract can relieve the congestion, swelling, hypertrophy and hyperplasia of the liver caused by obesity and reduce the degree of fatty liver. In addition, compared with a model control group, the coefficients of the three white fats, namely the inguinal fat, the epididymal fat and the perirenal fat, of mice in the low and high dose groups of the eucalyptus leaf extract are all obviously reduced, which reflects that the eucalyptus leaf extract can effectively reduce the accumulation of fat in the body and has the effect of losing weight.
The contents of total triglycerides, total cholesterol, low density lipoproteins, and high density lipoproteins in the mouse serum were measured, and the results are shown in FIGS. 2 to 5 below.
As can be seen from fig. 2 to 5, compared with the high-fat diet control group, the eucalyptus leaf extract significantly reduces total triglycerides, total cholesterol and low-density lipoproteins in the serum of the mice in the low and high dose groups, and significantly increases the HDL-C level in the serum (P < 0.01), which indicates that the eucalyptus leaf extract of the present invention can effectively improve the high-fat diet-induced lipid metabolism disorder of the body, and has a good lipid-lowering effect.
The above examples show that the eucalyptus leaf extraction method provided by the invention has high extraction rate, the obtained eucalyptus leaf extract has high content of total phenols, and the types of active substances are various. The eucalyptus leaf extract obtained by the preparation method can effectively inhibit the organism obesity caused by high-fat diet, has good weight-losing effect, can relieve the congestion, swelling and hypertrophy of the liver caused by the obesity, relieve the degree of fatty liver, effectively reduce the accumulation of fat in the organism, play a weight-losing effect, effectively improve the organism lipid metabolism disorder induced by the high-fat diet, and has good lipid-lowering effect.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. A preparation method of a eucalyptus leaf extract is characterized by comprising the following steps:
pulverizing folium Eucalypti Globueli to obtain folium Eucalypti Globueli powder;
performing continuous phase change extraction on the eucalyptus leaf powder to obtain an extracting solution;
sequentially concentrating and spray drying the extracting solution to obtain a crude extract of the eucalyptus leaves;
purifying the eucalyptus crude extract to obtain a eucalyptus extract;
the extractant adopted by the continuous phase-change extraction is butane, ethanol aqueous solution and water in sequence.
2. The method for producing a eucalyptus extract according to claim 1, wherein the particle size of the eucalyptus powder is 550 μm or less.
3. The method for preparing eucalyptus leaf extract as claimed in claim 2, wherein the extraction pressure of the continuous phase transition extraction is 0.5-1.5 MPa, the extraction temperature is 55-65 ℃, the resolution temperature is 65-75 ℃, and the extraction time of each extractant is 80-120 min.
4. The method for preparing a eucalyptus leaf extract as claimed in claim 3, wherein the purification is performed by silica gel column separation, and the silica gel column separation has a mesh size of 100-300 mesh.
5. The method for preparing the eucalyptus leaf extract, as claimed in claim 4, wherein the silica gel column separation process adopts gradient elution, and the elution flow rate of the gradient elution is 30-50 ml/min.
6. The method for preparing eucalyptus leaf extract as claimed in claim 5, wherein the eluent used for the silica gel column separation comprises component A and component B;
the component A is ethanol, and the component B is ethyl acetate or petroleum ether.
7. The preparation method of eucalyptus leaf extract according to claim 6, wherein during the gradient elution, the volume ratio of component A to component B is, in sequence, from (0);
eluting 2-4 column volumes of eluent with each volume ratio.
8. The method for preparing eucalyptus leaf extract as claimed in claim 7, wherein the eluted product is concentrated and spray-dried to obtain eucalyptus leaf extract.
9. The eucalyptus leaf extract prepared by the method according to any one of claims 1 to 8, which comprises 10 to 20% of oenothein B, 1 to 2% of terlimacin I, 1 to 1.5% of quercetin-3-O-glucuronide, 0.8 to 1.2% of gallic acid, 0.2 to 0.5% of terlimacin II, 3 to 6% of 1,2,3,4,6 pentagalloylglucose, wherein the content of total phenols in the eucalyptus leaf extract is 50 to 60%.
10. The use of the eucalyptus leaf extract of claim 9 for the preparation of a weight-reducing and lipid-lowering product.
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| JPH06133732A (en) * | 1992-10-28 | 1994-05-17 | Okinawa Yuukari Farm:Kk | Production of eucalyptus powder and eucalyptus extract |
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| JPH06133732A (en) * | 1992-10-28 | 1994-05-17 | Okinawa Yuukari Farm:Kk | Production of eucalyptus powder and eucalyptus extract |
| CN1406623A (en) * | 2001-09-11 | 2003-04-02 | 长冈香料株式会社 | Levulose absorption retardant, compound, food and levulose absorption repressor |
| JP2008163004A (en) * | 2006-12-08 | 2008-07-17 | Bhn Kk | Fat accumulation inhibitor |
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