CN115400165B - Eucalyptus extract, preparation method and application thereof in preparation of weight-losing and lipid-lowering products - Google Patents
Eucalyptus extract, preparation method and application thereof in preparation of weight-losing and lipid-lowering products Download PDFInfo
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- CN115400165B CN115400165B CN202211003304.4A CN202211003304A CN115400165B CN 115400165 B CN115400165 B CN 115400165B CN 202211003304 A CN202211003304 A CN 202211003304A CN 115400165 B CN115400165 B CN 115400165B
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- eucalyptus
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- eucalyptus extract
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Classifications
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A61K36/185—Magnoliopsida (dicotyledons)
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Mycology (AREA)
- Epidemiology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides a eucalyptus extract, a preparation method and application thereof in preparing a weight-losing and lipid-lowering product, and belongs to the technical field of plant extraction. The invention solves the problems of lower extraction rate of active ingredients of eucalyptus leaves, poorer activity of extracted products and the like in the prior art by adopting the processes of multi-solvent low-temperature continuous phase-change extraction, column chromatography separation and the like. The preparation method of the eucalyptus extract can realize higher impurity removal rate, and has simple operation steps and high yield. The eucalyptus 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 weight of a body, regulate the blood lipid level and effectively prevent and improve obesity. In addition, the eucalyptus resources are abundant and low in price, the preparation method is simple and low in cost, meets the industrial production requirements, has practical significance and application value in developing medicines or health-care products, and therefore the utilization rate of the eucalyptus resources is improved.
Description
Technical Field
The invention relates to the technical field of plant extraction, in particular to a eucalyptus extract, a preparation method and application thereof in preparing a weight-losing and lipid-lowering product.
Background
In recent years, with the increasing level of living, the incidence of nutritional obesity due to unhealthy work and eating patterns has increased. Nutritional obesity has been reported to increase the risk of developing hypertension, type ii diabetes, coronary heart disease and cancer, and to greatly affect body health. Therefore, how to effectively prevent and treat obesity has become one of the public safety and health problems worldwide. Weight loss drugs, while effective in controlling body weight, may have adverse effects on the body when taken for extended periods of time. Therefore, the market demand of developing safe weight-losing and lipid-lowering functional natural products without toxic or side effects is gradually released, and the future industry development prospect is good.
The eucalyptus resources of China are rich, and the eucalyptus tree has the characteristics of large area, wide region, multiple varieties and the like. Eucalyptus leaves are used as byproducts of eucalyptus and are always used as folk traditional Chinese herbal medicines for clearing heat, detoxicating, treating cold and the like. The prior researches show that the eucalyptus extract contains a large amount of polyphenols, has strong antioxidant activity in vivo and in vitro, and a series of effects of resisting aging, resisting tumor, resisting diabetes, resisting inflammation and the like are also reported, but the prior methods such as a chemical method, an enzymatic method and the like have lower extraction rate of the eucalyptus active ingredient, poorer activity of an extraction product and limit the application of the eucalyptus active ingredient in industry. At present, no report on the prevention and treatment of obesity by using eucalyptus extract is seen.
Disclosure of Invention
The invention aims to provide a eucalyptus extract, a preparation method and application thereof in preparing a weight-losing and lipid-lowering product, and solves the problems of low extraction rate of eucalyptus active ingredients, poor activity of the extracted product 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 eucalyptus extract, which comprises the following steps:
crushing eucalyptus leaves to obtain eucalyptus powder;
carrying out continuous phase-change extraction on eucalyptus powder to obtain an extracting solution;
concentrating and spray drying the extracting solution in turn to obtain a crude eucalyptus extract;
purifying the crude eucalyptus extract to obtain a eucalyptus extract;
the extractant adopted by the continuous phase-change extraction is butane, ethanol water solution and water in sequence.
Preferably, the particle size of the eucalyptus powder is 550 μm or less.
Preferably, the extraction pressure of the continuous phase-change extraction is 0.5-1.5 MPa, the extraction temperature is 55-65 ℃, the analysis temperature is 65-75 ℃, and the extraction time of each extractant is 80-120 min.
Preferably, the purification adopts silica gel column separation, and the mesh number of the silica gel column adopted by the 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, in the gradient elution process, the volume ratio of the component A to the component B is sequentially 0:100, 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, 100:0, and after the gradient elution is finished, collecting an elution product of which the volume ratio of the component A to the component B is 60:40, 70:30, 80:20;
eluting 2-4 column volumes with each volume ratio of eluent.
Preferably, the eluted product is concentrated and spray dried to obtain eucalyptus extract.
The invention also provides a eucalyptus extract prepared by the preparation method, which comprises 10-20% of evening primrose extract B, 1-2% of tricresazurin I, 1-1.5% of quercetin-3-O-glucuronide, 0.8-1.2% of gallic acid, 0.2-0.5% of tricresazurin II and 3-6% of 1,2,3,4,6 pentagalloylglucose, wherein the total phenol content of the eucalyptus extract is 50-60%.
The invention also provides application of the eucalyptus extract in preparing a weight-losing and lipid-lowering product.
The invention has the technical effects and advantages that:
the preparation method of the eucalyptus extract can realize higher impurity removal rate, and has simple operation steps and high yield. The eucalyptus extract obtained by the preparation method has higher polyphenol content and higher total yield, and is suitable for being applied to the actual industry. The eucalyptus extract obtained by the method has good weight-losing and lipid-lowering effects, can effectively control the weight of a body, regulate the blood lipid level and effectively prevent and improve obesity. In addition, the eucalyptus resources are abundant and low in price, the preparation method is simple and low in cost, meets the industrial production requirements, has practical significance and application value in developing medicines or health-care products, and improves the utilization rate of the eucalyptus resources to a certain extent.
Drawings
FIG. 1 shows the results of liquid chromatography;
FIG. 2 shows the results of the total triglyceride content measurement in the serum of mice;
FIG. 3 shows the results of determination of total cholesterol content in mouse serum;
FIG. 4 shows the results of the measurement of the low density lipoprotein content in the serum of mice;
FIG. 5 shows the results of measurement of the content of high density lipoprotein in serum of mice.
Detailed Description
The invention provides a preparation method of eucalyptus extract, which comprises the following steps:
crushing eucalyptus leaves to obtain eucalyptus powder;
carrying out continuous phase-change extraction on eucalyptus powder to obtain an extracting solution;
concentrating and spray drying the extracting solution in turn to obtain a crude eucalyptus extract;
purifying the crude eucalyptus extract to obtain a eucalyptus extract;
the extractant adopted by the continuous phase-change extraction is butane, ethanol water solution and water.
In the invention, the eucalyptus leaves are preferably crushed after being dried in shade to obtain eucalyptus powder, and the particle size of the eucalyptus powder is preferably below 550 μm, and more preferably below 350 μm; carrying out continuous phase-change extraction on eucalyptus powder, wherein the continuous phase-change extraction is preferably carried out by adopting a continuous phase-change extraction kettle, the extraction pressure of the continuous phase-change extraction is preferably 0.5-1.5 MPa, more preferably 0.8-1.2 MPa, the extraction temperature is preferably 55-65 ℃, more preferably 58-62 ℃, the analysis temperature is preferably 65-75 ℃, more preferably 68-72 ℃, and the extraction time of each time is preferably 80-120 min, more preferably 90-110 min; the continuous phase-change extraction adopts butane, ethanol water solution and water as the extracting agents, the extraction sequence is not required, and the volume concentration of the ethanol water solution is preferably 60-80%.
In the present invention, the concentration is preferably 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 atomizer frequency is preferably 40-50 Hz.
In the present invention, the purification is preferably performed by a silica gel column separation, and the mesh number of the silica gel column used in the silica gel column separation is preferably 100 to 300 mesh, and more preferably 150 to 250 mesh; in the silica gel column separation process, gradient elution is preferably adopted, and the elution flow rate of the gradient elution is preferably 30-50 ml/min, and more preferably 35-45 ml/min; the eluent used for 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; in the gradient elution process, the volume ratio of the component A to the component B is preferably 0:100, 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10 and 100:0 in sequence, and after the gradient elution is finished, the elution products of which the volume ratio of the component A to the component B is 60:40, 70:30 and 80:20 are collected; in the invention, the eluent with each volume ratio preferably elutes 2-4 column volumes, after gradient elution is finished, the elution product is preferably concentrated and spray-dried to obtain the eucalyptus extract, the concentration is preferably vacuum concentration, the vacuum concentration temperature is preferably 65-85 ℃, the vacuum degree is preferably 400-600 Pa, the air inlet temperature of spray-drying is preferably 250-300 ℃, the feeding speed is preferably 100-150 rpm, and the atomizer frequency is preferably 50-60 Hz.
The invention also provides the eucalyptus extract prepared by the preparation method.
The invention also provides application of the eucalyptus 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 contains the eucalyptus extract of the invention as the only active ingredient; the product preferably further comprises auxiliary materials, the types of the auxiliary materials are not particularly limited, and the auxiliary materials of the product are conventional in the art; the product formulation 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, lubricant, disintegrant, cosolvent, diluent, stabilizer, suspending agent, pigment, flavoring agent, etc. for oral preparation; preservatives, solubilizing agents, stabilizers and the like for injectable formulations; matrix 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
Drying eucalyptus leaves of Guanglin No. 9 in shade, mechanically pulverizing to 30 meshes, weighing eucalyptus powder 20kg, loading into a continuous phase-change extraction kettle, sealing and extracting at 65 deg.C and 75 deg.C under 1.0MPa, sequentially extracting with butane, 70% ethanol and water for 100min to obtain eucalyptus extract. Concentrating the eucalyptus leaf extract under vacuum at 60deg.C and 400Pa, and spray drying at 200deg.C and feed rate of 100rpm and atomizer frequency of 45Hz to obtain crude extract. The crude eucalyptus leaf extract is subjected to 100-mesh silica gel column, elution is carried out at a flow rate of 40ml/min, gradient elution is carried out by ethanol and ethyl acetate, the volume ratio of the ethanol to the ethyl acetate is 0:100, 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10 and 100:0, 3 column volumes are eluted according to each ratio, the eluent with the volume ratio of the ethanol to the ethyl acetate of 60:40, 70:30 and 80:20 is concentrated in vacuum under the condition of 70 ℃ and 500Pa, and spray drying is carried out at the air inlet temperature of 250 ℃ and the feed speed of 100rpm and the atomizer frequency of 50Hz, so that the eucalyptus leaf extract is obtained.
Example 2
Drying eucalyptus leaf raw material of Guanglin No. 9 in shade, mechanically pulverizing to 40 meshes, weighing 15kg of eucalyptus leaf powder, loading into a continuous phase-change extraction kettle for sealed extraction, wherein the extraction pressure is 1.2MPa, the extraction temperature is 60 ℃, the analysis temperature is 70 ℃, butane, 65% ethanol and water are sequentially used as extracting agents, the extraction time is 120min each time, and collecting eucalyptus leaf extract. Concentrating folium Eucalypti Globueli extractive solution under reduced pressure at 65deg.C and 500Pa, and spray drying at 250deg.C with feed rate of 150rpm and atomizer frequency of 50Hz. The crude eucalyptus leaf extract is subjected to 200-mesh silica gel column, elution is carried out at a flow rate of 35ml/min, gradient elution is carried out by ethanol and ethyl acetate, the volume ratio of the ethanol to the ethyl acetate is 0:100, 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10 and 100:0, 3 column volumes are eluted according to each ratio, the eluent with the volume ratio of the ethanol to the ethyl acetate of 60:40, 70:30 and 80:20 is concentrated in vacuum under the condition of 70 ℃ and 500Pa, and spray drying is carried out at the air inlet temperature of 300 ℃, the feeding speed of 150rpm and the atomizer frequency of 60Hz, so that the eucalyptus leaf extract is obtained.
Example 3
Drying eucalyptus leaf raw material of Guanglin No. 9 in shade, mechanically pulverizing to 60 meshes, weighing 15kg of eucalyptus leaf powder, loading into a continuous phase-change extraction kettle for sealed extraction, wherein the extraction pressure is 1.2MPa, the extraction temperature is 65 ℃, the analysis temperature is 75 ℃, butane, 80% ethanol and water are respectively used as extracting agents, the extraction time is 120min each time, and collecting eucalyptus leaf extract. Concentrating the eucalyptus leaf extract under vacuum at 60deg.C and 400Pa, and spray drying at 200deg.C and feed rate of 100rpm and atomizer frequency of 45Hz to obtain crude extract. The crude eucalyptus leaf extract is subjected to 200-mesh silica gel column, elution is carried out at a flow rate of 35ml/min, gradient elution is carried out by ethanol and petroleum ether, the volume ratio of the ethanol to the petroleum ether is 0:100, 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10 and 100:0 in sequence, 3 column volumes are eluted in each ratio, the eluent with the volume ratio of 60:40, 70:30 and 80:20 is concentrated in vacuum under the condition of 70 ℃ and 500Pa, and spray drying is carried out under the conditions of the air inlet temperature of 250 ℃, the feeding speed of 100rpm and the atomizer frequency of 50Hz, so that the eucalyptus leaf extract is obtained.
Experimental example 1 liquid chromatography
The eucalyptus extract obtained in example 1 was subjected to liquid chromatography under the conditions of a Diamonsil C18 column (250X 4.6mm,5 μm); the detection wavelength is 270nm; the mobile phase is 10% -90% methanol for gradient elution for 1h, and the flow rate is 1 mL/mm; the sample injection amount was 20. Mu.L.
The results are shown in fig. 1, wherein: 1. gemin D; 2. evening primrose element B; 3. 5-methoxy-aldol; 4. quercus robusta ellagic acid; 5. tricin I; 6. chlorogenic acid; 7. tricin II; 8. 1,2,3, 6-tetragalloylglucose; 9. ethyl gallate; 10. 1,2,3,4, 6-pentagalloylglucose; 11. quercetin-3-O-glucuronide.
Experimental example 2 determination of extraction Rate and Total phenol content
The eucalyptus extract mass in example 1 and example 2 was measured, and the extraction rate (extraction rate=eucalyptus extract mass/eucalyptus powder mass×100%) was calculated;
the total phenol content was measured by the Fu Lin Fen method and the results are shown in Table 1 below:
TABLE 1 extraction results of Eucalyptus extract
| Quality of extract of eucalyptus leaves | Extraction rate | Total phenol content | |
| Example 1 | 11.2kg | 56％ | 55％ |
| Example 2 | 9.0kg | 60％ | 50％ |
Experimental example 3 study of fat-reducing and fat-reducing effects
Test animals: male, SPF grade C57BL/6J mice, weighing approximately (23+ -2 g), purchased from Peking Vitrenia laboratory animal technologies Co., ltd;
test drugs and reagents: eucalyptus extract with a total phenol content of about 55%; physiological saline; kits for the same; kits for Total Cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL-C), high density lipoprotein (HDL-C), and the like;
feed: 60% high-fat model feed and 10% low-fat control feed were purchased from south Tongton Talofei feed technologies Co., ltd;
the experimental method comprises the following steps: after 7d adaptation, 32C 57BL/6J mice were divided into four groups, which were normal control group, model control group, eucalyptus extract low dose group (50 mg/kg) and eucalyptus extract high dose group (100 mg/kg), respectively. The normal control group was fed with 10% low fat control feed, while the model group and the low and high dose eucalyptus extract group were fed with 60% high fat model feed for a period of 10 weeks. Beginning at 1d, normal control group and model control group mice were perfused with normal saline, and low and high dose eucalyptus extract groups mice were perfused with 50mg/kg and 100mg/kg eucalyptus extract, respectively. After the feeding period is completed, the mice are sacrificed, and serum, viscera and fat are collected and stored at-80 ℃ for testing.
The body weight, food intake, organ factor, fat factor, serum lipid level, and the like of the mice were measured in the experiments, and the results are as follows. All experimental data were analyzed using SPSS 17.0 statistical software. The comparison between groups uses One-way analysis of variance (One-wayAnova), and the comparison between groups uses Duncan method. Results are expressed as mean ± standard deviation, p <0.05 being the difference significant.
During the test period, the body weights of the mice in each group were weighed and recorded weekly, and the body weights and body weight gain of each group are shown in table 2.
TABLE 2 weight gain (g) in mice
Note that: data in the table are mean value of samples ± standard deviation; p <0.01 compared to normal control group, and # p <0.05 compared to model control group.
As can be seen from table 2, the weight gain rate of the mice in the model control group is significantly higher than that of the mice in the normal control group, which indicates that the establishment of the obesity model of the mice is successful. Compared with the model control group, the weight gain of mice in the group with low eucalyptus extract and high dosage is reduced, wherein the weight gain of the mice in the group with high dosage is extremely remarkable. The result shows that the eucalyptus extract with a certain dosage can effectively inhibit the body obesity caused by high-fat diet, and has good weight-losing effect.
The epididymal fat, inguinal fat, perirenal fat, brown fat and liver of each group of mice were weighed and their indices were calculated, and the results are shown in table 3.
TABLE 3 liver and fat coefficients of mice (%)
Note that: data in the table are mean value of samples ± standard deviation; p <0.01 compared to normal control group, and # p <0.05 compared to model control group.
As can be seen from table 3, the liver coefficients of the mice in the low and high dose groups of eucalyptus extract are significantly lower than those of the mice in the model control group, which indicates that the eucalyptus extract can relieve liver congestion, swelling, hypertrophy and hyperplasia caused by obesity and reduce the degree of fatty liver. In addition, compared with a model control group, the white fat coefficients of the three kinds of fat, namely the inguinal fat, the epididymal fat and the perirenal fat of the mice in the low-dose and high-dose groups of eucalyptus extract are obviously reduced, and the effect that the eucalyptus extract can effectively reduce the accumulation of fat in the body of the organism and has the effect of losing weight is reflected.
The results of measuring the total triglyceride, total cholesterol, low density lipoprotein and high density lipoprotein content in the serum of mice are shown in FIGS. 2 to 5 below.
As shown in fig. 2 to 5, compared with the high-fat diet control group, the eucalyptus extract has significantly reduced total triglyceride, total cholesterol and low-density lipoprotein in the serum of the mice in the low-and high-dose group, and the HDL-C level in the serum is significantly increased (P < 0.01), which indicates that the eucalyptus extract of the invention can effectively improve lipid metabolism disorder of the organism induced by the high-fat diet, and has good lipid-lowering effect.
As can be seen from the above examples, the eucalyptus extraction method provided by the invention has high extraction rate, and the obtained eucalyptus extract has high total phenol content and various active substances. The eucalyptus extract obtained by the preparation method disclosed by the invention can effectively inhibit body obesity caused by high-fat diet, has a good weight-reducing effect, can relieve liver congestion, swelling, hypertrophy and hyperplasia caused by obesity, lighten the degree of fatty liver, can effectively reduce fat accumulation in a body, has the weight-reducing effect, can effectively improve lipid metabolism disorder of the body induced by high-fat diet, and has a good lipid-lowering effect.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (2)
1. The application of the eucalyptus extract in preparing the weight-losing and lipid-lowering medicines is characterized in that the preparation method of the eucalyptus extract comprises the following steps:
crushing eucalyptus leaves to obtain eucalyptus powder;
carrying out continuous phase-change extraction on eucalyptus powder to obtain an extracting solution;
concentrating and spray drying the extracting solution in turn to obtain a crude eucalyptus extract;
purifying the crude eucalyptus extract to obtain a eucalyptus extract;
the extractant adopted by the continuous phase-change extraction is butane, ethanol water solution and water in sequence;
the purification adopts silica gel column separation, and the mesh number of the silica gel column adopted by the silica gel column separation is 100-300 meshes;
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;
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;
in the gradient elution process, the volume ratio of the component A to the component B is sequentially 0:100, 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10 and 100:0, and after the gradient elution is finished, the elution products of which the volume ratio of the component A to the component B is 60:40, 70:30 and 80:20 are collected;
eluting 2-4 column volumes by using eluent with each volume ratio;
concentrating and spray drying the eluted product to obtain eucalyptus extract;
the particle size of the eucalyptus powder is below 550 mu m;
the extraction pressure of the continuous phase-change extraction is 0.5-1.5 MPa, the extraction temperature is 55-65 ℃, the analysis temperature is 65-75 ℃, and the extraction time of each extractant is 80-120 min.
2. The use according to claim 1, wherein the eucalyptus extract comprises 10 to 20% of evening primrose extract B, 1 to 2% of triemic acid I, 1 to 1.5% of quercetin-3-O-glucuronide, 0.8 to 1.2% of gallic acid, 0.2 to 0.5% of triemic acid II, 3 to 6% of 1,2,3,4, 6-pentagalloylglucose, and the total phenol content of the eucalyptus extract is 50 to 60%.
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| JP2008163004A (en) * | 2006-12-08 | 2008-07-17 | Bhn Kk | Fat accumulation inhibitor |
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