CN116139191A - Preparation method and application of mangosteen extract - Google Patents
Preparation method and application of mangosteen extract Download PDFInfo
- Publication number
- CN116139191A CN116139191A CN202310109986.5A CN202310109986A CN116139191A CN 116139191 A CN116139191 A CN 116139191A CN 202310109986 A CN202310109986 A CN 202310109986A CN 116139191 A CN116139191 A CN 116139191A
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- mangosteen extract
- mangosteen
- ethanol
- extract
- extraction
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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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- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
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- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
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Abstract
The invention discloses a preparation method and application of mangosteen extract. Compared with the mangosteen extract prepared by pure water extraction and alcohol extraction, the method has the advantages that the extraction rate of alpha-mangostin and total polyphenol in the mangosteen extract is obviously improved, and the mangosteen extract has better antibacterial activity on sensitive staphylococcus aureus, methicillin-resistant staphylococcus aureus, vancomycin-resistant enterococci and clinically separated clostridium perfringens, and in addition, the extract contains more total polyphenol and has the effect of resisting oxidization, and the aim of promoting animal growth is fulfilled by maintaining animal intestinal integrity and health.
Description
Technical Field
The invention relates to the technical fields of chemistry, agriculture and medicine, in particular to a preparation method and application of a mangosteen extract.
Background
The Chinese is the livestock breeding industry and the large country of animal source food production, consumption and trade, especially the pig and chicken breeding amount is huge, and the safety of meat, eggs and milk is relevant to the health of the common people. Bacterial infection seriously threatens the health of livestock and poultry farming, causing huge economic loss (Nature, 2016,535,85). Antibacterial agents remain the first choice for the treatment of bacterial infectious diseases. To date, bacterial resistance has rapidly emerged worldwide, and the development of alternatives to feed antibiotics such as plant extracts has become a current research and development hotspot (AdvancedScience, 2021,8,2100749;RSCMedicinal Chemistry,2021,13,107).
Mangosteen (Garcinia mangostana) is a plant of the genus garcinia of the family garcinia, a tropical fruit native to southeast asia, and has been known for its medicinal and health-promoting properties (Foodand Chemical Toxicology,2017,109,102). Mangosteen pericarp has been widely used as a traditional medicine in southeast asia as early as several hundred years ago for the treatment of diarrhea, abdominal pain, skin infections, infectious wounds, chronic ulcers, malaria, sepsis, leukemia, etc. (Journal of AgriculturalandFood Chemistry,2015, 63,7670). Early researches show that the mangosteen pericarp extract has the effects of resisting bacteria, diminishing inflammation, resisting oxidation and promoting growth, and the aim of promoting the growth of animals is fulfilled by maintaining the integrity and health of animal intestinal tracts. Meanwhile, the mangosteen extract is not only applied to the field of natural medicines, but also applied to a plurality of fields such as health products, cosmetics, veterinary medicines and the like, has very wide application fields and broad market prospect, and has higher research and development values.
In recent years, mangosteen extract has been a research hotspot due to its excellent antibacterial effect, readily available raw materials, simple extraction process, and the like, and many patent documents on mangosteen extract are currently disclosed. The patent CN103467433A and the patent CN101983722A adopt a method of refluxing with deionized water to remove water-soluble components with larger polarity, thereby enriching the active components of mangosteen pericarp; the patent CN114392214A adopts a normal hexane dipping method to remove less polar components such as fatty acid and the like, and then adopts ethanol reflux extraction to obtain the mangosteen extract. The method has the advantages of simple extraction process, simple operation, high yield and high quality of mangosteen extract, and has the disadvantages of relatively long time consumption and large use amount of organic reagent. In order to shorten the extraction time, patent CN109362967a adopts an ultrasonic extraction method. The advantage of this method is that the extraction time is significantly reduced, but special instrumentation is required. The patent CN109998110A and the patent CN109938232A are extracted by adopting an enzymolysis method, and any organic reagent is not used in the extraction process, so that the safety of the mangosteen extract is improved. But the cost of the feed additive is high, and the use of the feed additive is not recommended. The patents CN101904880a and HK1165320a used a heated reflux method, using a mixed reagent for extraction. The method has simple process, but low product quality and content.
The prior art has the problems of complex process, large dosage of organic reagent, long operation time, high cost, low yield and the like. In view of this, there is a need to develop a new extraction method to solve the above-mentioned technical inadequacies. In fact, besides alpha-mangostin, a large amount of polyphenols, flavonoids and polysaccharides (Contemporary ClinicalDentistry,2019, 10,123) exist in the mangosteen pericarp, and active substances are extracted as much as possible through optimization of an extraction process, so that the utilization value of the mangosteen pericarp is improved, and the mangosteen pericarp is expected to be used as a feed additive for replacing antibiotic functions and applied to livestock and poultry farming.
Disclosure of Invention
The invention aims to provide a preparation method of a mangosteen extract which is simple to operate, low in reagent consumption, low in cost, capable of being produced in a large scale and rich in alpha-mangostin, and meanwhile, total polyphenol in mangosteen pericarp is extracted, and the extract is used in the field of feed additives and is used as an antibacterial, antioxidant and growth-promoting plant extract in the field of feed additives.
The preparation method of the mangosteen extract provided by the invention comprises the following steps: adding any one of the extraction solvents 1) -3) into mangosteen pericarp powder, heating and reflux-extracting, filtering, mixing filtrates, concentrating under reduced pressure until no alcohol smell exists, and lyophilizing to obtain mangosteen extract;
1) An aqueous ethanol solution in which the volume concentration of ethanol is 50 to 90%, preferably 70 to 90%, 80 to 90%, more preferably 80%;
2) The heating reflux extraction is carried out in two steps, wherein the extraction solvent used in the first step is 0.5-1 per mill formic acid aqueous solution, and the extraction solvent used in the second step is ethanol aqueous solution, wherein the volume concentration of ethanol is 50-90%, preferably 70-90%, more preferably 80-90%;
3) The mixed solvent of ethanol, ethyl acetate and water, wherein the volume concentration of the ethanol is 45-90%, the volume concentration of the ethyl acetate is 0-50%, preferably 0-45%, and the water balance, specifically the volume ratio of the ethanol, the ethyl acetate and the water can be: 45-90: 0 to 45:10 to 30, preferably 45 to 60: 30-45: 10, more preferably 60:30:10;
the ratio of the mangosteen pericarp powder to the extraction solvent can be 1g:8 to 15L of the total weight of the product,
the temperature of the reflux can be 100-140 ℃ and the time can be 0.5-2.5 hours;
the heating reflux extraction can be carried out for a plurality of times, and can be carried out for 1 to 5 times in particular;
the conditions of the decompression concentration can be 40-70 ℃ and minus 0.07-minus 0.08MPa;
the method further comprises adding adjuvants into the obtained mangosteen extract, and spray drying to obtain powder mangosteen extract rich in alpha-mangostin and total polyphenols.
The auxiliary materials comprise: maltodextrin, corn starch, sucrose, anhydrous dextrose, and the like.
The mangosteen pericarp powder is prepared by the following steps: selecting fresh mangosteen fruit with dark red skin, cleaning, removing edible pulp, cutting pericarp into small pieces, drying in oven at 45deg.C for 72 hr, grinding the dried pericarp into fine powder, sieving with 20 mesh sieve, and storing the obtained powder in a sealed light-proof container.
The mangosteen extract prepared by the method also belongs to the protection scope of the invention.
The infrared spectrum of the mangosteen extract is 3400cm -1 Characteristic absorption peak with hydroxyl group at about 3000-2850cm -1 The region has C-H telescopic vibration absorption peak of 1600cm -1 About 1300-1000cm with characteristic absorption peak of benzene ring -1 Has C-O telescopic vibration absorption peak;
the mangosteen extract mainly comprises alpha-mangostin and total polyphenol.
The mangosteen extract is used for preparing veterinary medicines or feed additives related to antibiosis, anti-inflammation, antioxidation and growth promotion in livestock and poultry cultivation;
the livestock and poultry comprise pigs, chickens, cattle and the like.
The veterinary drug or the drug feed additive is administered orally or by feed mixing.
The mangosteen extract can be used as veterinary medicine or medicine feed additive in the form of liquid preparation, powder or premix.
In particular, the antibacterial is against gram-positive bacteria,
the gram positive bacteria include, but are not limited to, sensitive staphylococcus aureus, methicillin-resistant staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), and clostridium perfringens.
The mangosteen extract is derived from natural plants, has simple extraction process, short production period and low cost, adopts the mangosteen epicarp as the raw material, belongs to waste recycling, and is low in cost and easy to obtain. Compared with the mangosteen extract prepared by pure water extraction and alcohol extraction, the method has the advantages that the extraction rate of alpha-mangostin and total polyphenol in the mangosteen extract is obviously improved, and the mangosteen extract has better antibacterial activity on sensitive staphylococcus aureus, methicillin-resistant staphylococcus aureus (MRSA), vancomycin-resistant enterococcus (VRE) and clinically separated clostridium perfringens gram positive bacteria, and in addition, the extract contains more total polyphenol and has the effect of resisting oxidization, and the aim of promoting animal growth is fulfilled by maintaining animal intestinal integrity and health. Therefore, the product has higher development and research values as veterinary drugs or feed additives, and has wide application prospects in animal health care and livestock and poultry cultivation.
Drawings
Fig. 1 is an infrared spectrum of the mangosteen extract in example 1 of the present invention.
FIG. 2 is a graph showing the analysis of the components of the mangosteen extract prepared in examples 2-4 according to the different extraction methods of the present invention.
FIG. 3 is a graph showing the growth of Staphylococcus aureus ATCC 29213 of example 6 of the present invention at various concentrations of mangosteen extract. (A) Inhibition of growth of staphylococcus aureus ATCC 29213 by AMG standard at different concentrations; (B) Inhibition of staphylococcus aureus ATCC 29213 growth by mangosteen extract at different concentrations.
FIG. 4 is a graph showing the antioxidant capacity of the Carcinia Maultflora champ extract of example 7 of the present invention. (a) the action of mangosteen extract on DPPH; (B) Mangosteen extract ABTS + Cleaning ability; (C) Measurement of the reducing ability of mangosteen extract to iron ions.
FIG. 5 shows the measurement of the total polyphenol content of the mangosteen extract in example 8 of the present invention.
Detailed Description
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the invention in any way.
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
The strain used in the test: staphylococcus aureus ATCC 29213 was purchased from american type culture collection (American type culture collection, ATCC) and kept in the laboratory; clostridium perfringens of chicken source a CVCC 2030 and clostridium perfringens of chicken source C CVCC 60102 were purchased from the chinese veterinary microbiological bacterial deposit management center of the chinese veterinary drug administration; the rest clinical drug-resistant strains are obtained by separating in the laboratory and are stored in the laboratory.
Main reagent and consumptive material: MH Medium (MHB) was purchased from beijing land bridge technology division, liability company; alpha-mangostin was purchased from the scientific development limited of the method Dou Purui.
Example 1 Infrared Spectroscopy of mangosteen extract
Preparation of the samples: potassium bromide was first dried in an oven for a period of time to remove as much moisture as possible from the mixture. 2mg of N-heterocyclic carbene Nickel Complex [ Ni (NHC) was added to an agate mortar](PF 6 ) 2 And 150mg of dry potassium bromide powder, and grinding the two with a grinding rod, mixing uniformly, and grinding thoroughly. Then pouring small amount of mangosteen extract (commercially available) powder into infrared grinding tool, covering with cover, compacting, and tabletting. After pressing, the sample is placed on a rack for holding the sample.
The measuring method comprises the following steps: prior to testing the sample, a blank, i.e., a sheet pressed with potassium bromide, was scanned to subtract the effect of potassium bromide on the spectrum of the sample being tested. Placing the above frame fixed with sample on a sample frame in an infrared spectrometer sample chamber at 450-4000cm -1 Infrared spectral scans are performed in the range.
Example 2 mangosteen extract extraction Process one
Weighing 100g of mangosteen pericarp dry powder, adding 8 times volume of acid aqueous solution (0.5-1%o formic acid (0.4-0.8 mL), v/v), and reflux-extracting at 120 ℃ for 2 hours. Filtering, extracting the filter residue with 8 times of 60-90% (v/v) ethanol water solution, refluxing at 120 ℃ for 2 hours, filtering for 2 times, and combining the filtrates. Evaporating with rotary evaporator until no alcohol smell, transferring into evaporating dish, and lyophilizing to obtain mangosteen extract.
Investigation indexes: (1) the paste yield is as follows: taking the extract yield as a secondary detection index (weight coefficient 0.3); (2) alpha-mangostin: the alpha-mangostin in the product is a substance which mainly plays an antibacterial activity, so that the content of the alpha-mangostin in the extract is used as a main detection index (weight coefficient 0.4) to perform optimization of an orthogonal process; (3) total polyphenols: the total polyphenol in the product has the effects of resisting inflammation and oxidation, is an effective component group of the product, and is a secondary detection index (weight coefficient 0.3).
Table 1 extraction process-table of experimental results of different extraction solvents
As can be seen from the data in table 1, the optimal parameters for the first mangosteen extract extraction process are: firstly, weighing 100g of mangosteen pericarp dry powder, adding 8 times of acid aqueous solution (0.5-1%o formic acid (0.4-0.8 mL), v/v), and extracting at 120 ℃ under reflux for 2 hours. Filtering, extracting the residue with 8 times of 80% (v/v) ethanol water solution, refluxing at 120deg.C for 2 hr, 2 times, filtering, and mixing filtrates. Evaporating with rotary evaporator until no alcohol smell, transferring into evaporating dish, and lyophilizing to obtain mangosteen extract.
Example 3 mangosteen extract extraction Process two
100g of mangosteen pericarp dry powder is weighed, 8 times of mixed solution (ethanol, ethyl acetate and water, V/V/V) is added, reflux is carried out at 120 ℃ for 2 hours, 2 times, filtration is carried out, and the filtrates are combined. Evaporating with rotary evaporator until no alcohol smell, transferring into evaporating dish, and lyophilizing to obtain mangosteen extract.
Table 2 extraction process two different extraction solvent experimental results table
Example 4 mangosteen extract extraction Process III
Weighing 100g of mangosteen pericarp dry powder, adding 8 times volume of ethanol water solution, refluxing at 120 ℃ for 2 hours for 2 times, filtering, and combining filtrates. Evaporating with rotary evaporator until no alcohol smell, transferring into evaporating dish, and lyophilizing to obtain mangosteen extract.
Table 3 extraction process three different extraction solvent experimental results table
Comparing the characteristic patterns of different extraction methods, the main components of the three optimization methods are similar. However, since the water-soluble fraction was not removed in the second and third schemes, a more polar component was present in the crude extract fraction (shown in FIG. 2).
By comparing the yields of the extracts of different extraction methods, we find that the three schemes have higher paste yield, the liquid absorption of the medicinal materials is 2 times, and the content of alpha-mangostin reaches more than 20 percent. Because the effective component of the mangosteen extract which plays an antibacterial role is mainly alpha-mangostin, the alpha-mangostin is used as an important evaluation index of an extraction process, and by comparing the content of the alpha-mangostin in different extraction methods, the purity of the alpha-mangostin in the scheme one is the highest, and the extraction amount of the alpha-mangostin in the scheme two is the highest under the condition of the same prescription amount. Meanwhile, the total polyphenol content of the first scheme and the second scheme is 220.47-227.26mg GAE/g DW, and the extraction rate is high.
TABLE 4 yields of mangosteen extract from different extraction methods
Example 5 determination of in vitro antibacterial Activity of mangosteen extract
The Minimum Inhibitory Concentration (MIC) assay of clinical chicken-derived clostridium perfringens was performed on alpha-mangostin and its derivatives with reference to the micro broth dilution method recommended by the american clinical laboratory standardization committee (CLSI 2022), and MIC result judgment was mainly performed with reference to the CLSI M100-32st Edition (2022) standard.
The step of detecting the minimum inhibitory concentration is as follows:
a) Drug-fold ratios were diluted to 10 concentration gradients using MHB medium and 100 μl/well was added sequentially to the first 10 columns of wells of a 96-well U-plate.
b) The concentration of the strain to be tested was about 1X 10 by dilution with MHB medium 6 CFU/mL, 100. Mu.L/well was added to the wells to which the drug had been added. As a negative control, 100. Mu.L/well of MHB medium was added to the 11 th well, and as a positive control, 100. Mu.L/well of bacterial liquid was added to the 12 th well.
c) After 18h incubation at 37 ℃, the reading is observed, and the minimum concentration inhibiting bacterial growth is the MIC value of the drug. One repeat was set for each test well.
We have determined the antimicrobial activity of the mangosteen extract against gram-positive and negative bacteria, as shown in Table 5, and found that the mangosteen extract has better antimicrobial activity against gram-positive bacteria including sensitive Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci and clinically isolated Clostridium perfringens, with a minimum inhibitory concentration of 2-8 μg/mL, and no antimicrobial activity against gram-negative bacteria (MIC >128 μg/mL). The results show that the mangosteen extract has excellent effect on various tested gram-positive bacteria and has no activity on gram-negative bacteria.
Table 5 determination of mangosteen extract antibacterial spectrum
EXAMPLE 6 growth curves of Staphylococcus aureus ATCC 29213 in mangosteen extract
a) Staphylococcus aureus ATCC 29213 was selected and cultured in MHB broth medium at 37℃and 200rpm in a shaker to logarithmic phase. Bacteria turbidity was adjusted to 0.5 with a turbidimeter and diluted 100-fold with MH broth medium (about 1.0 x 10) 6 CFUs/mL).
b) The drug to be tested is diluted to the required concentration by MH broth culture medium, 100 mu L is added into 96-well flat bottom plate, and 100 mu L of diluted bacteria liquid to be tested is added into each well. A negative control containing MH broth alone and a positive control containing the test bacterial fluid was set.
c) Setting the temperature of the enzyme-labeled instrument system to 37 ℃ and detecting OD 600nm Absorbance at this point was measured once per hour for a total of 24 hours. Each treatment was set up with 3 biological replicates.
As shown in FIG. 3, the content of alpha-mangostin in the mangosteen extract is 21.17%, and the active ingredient of the mangosteen extract, which has an antibacterial effect, is found to be alpha-mangostin by comparing the concentration of alpha-mangostin (2 mug/mL) with the concentration of the mangosteen extract (10 mug/mL) for completely inhibiting the growth of staphylococcus aureus.
Example 7 antioxidant Activity of mangosteen extract
Heat stress is an important factor affecting the productivity of poultry. Research shows that the heat stress of livestock and poultry firstly reduces the feed intake and the absorption of nutrient substances, thereby affecting the metabolism level of the organism and causing the oxidation resistance dysfunction of the organism, so that excessive free radicals are generated in the organism, which indicates that the heat stress can cause the oxidation stress of the organism.
The best approach currently considered to relieve oxidative stress is to use a variety of exogenous antioxidants. Therefore, we next evaluated the antioxidant activity of mangosteen extract in a number of ways.
(1) DPPH method for measuring antioxidant activity of mangosteen extract
DPPH is also called 1, 1-diphenyl-2-trinitrophenylhydrazine, is a very stable free radical of nitrogen center, and its stability mainly comes from steric hindrance of 3 benzene rings with resonance stabilization effect, so that unpaired electrons on nitrogen atom clamped in the middle cannot exert their due electron pairing effect. The absolute ethanol solution of the fluorescent dye has purple color, has maximum absorption at 517nm wavelength, and has linear relation between absorbance and concentration. When a radical scavenger is added thereto, DPPH may be combined or replaced, so that the number of radicals is reduced, absorbance is decreased, and the color of the solution is made lighter, whereby the ability to scavenge radicals can be evaluated. I.e. the antioxidant capacity was calculated by measuring the effect of the sample on DPPH removal at 517 nm.
(2) Abts method for measuring antioxidant activity
The assay is based on the radical scavenging activity of ABTS radical decolorization measurement reagents. Briefly, ABTS radical cations are synthesized by ABTS and potassium persulfate (K 2 S 2 O 8 ) The reaction in water occurs. The ABTS radical solution was diluted to an absorbance of 0.7±0.2 at 750nm and stored in the dark for 12 hours. Next, the ethanol extract was added to the ABTS radical solution. After 5 minutes, the absorbance of the mixture was measured using a microplate reader. Vitamin E and BHT were used as positive controls and trolox was used for calibration. The antioxidant potential is expressed as trolox equivalent antioxidant capacity (TEAC) value.
(3) FRAP method for measuring antioxidant activity
The measurement measures the reduction characteristics of the antioxidant based on the reduction of iron ions. Briefly, the FRAP reagent is prepared by mixing TPTZ and FeCl in acetate buffer 3 Solutions. The extract dissolved in ethanol was mixed with FRAP reagent in 96-well plates and allowed to react for 5 minutes at room temperature. Vitamin E and BHT were used as positive controls and ferrous sulfate was used for calibration. The absorbance of the resulting mixture was measured using a microtiter plate reader. The antioxidant activity of the extracts is expressed in Equivalent Capacity (EC) and is calibrated using ferrous sulfate. Higher EC value means higher reducing activity of the extract.
The results are shown in FIG. 4, and the determination of DPPH, ABTS free radical scavenging rate and Fe of mangosteen extract 2+ The mangosteen extract has a strong antioxidant capacity.
Example 8 determination of Total polyphenol content in mangosteen extract
The total content of phenolic compounds in the mangosteen extract is determined by the colorimetric Folin-Ciocalteu method. Briefly, the obtained extract samples were diluted in ethanol and pipetted into 96-well plates. Next, folin-Ciocalteu reagent and sodium carbonate were added to each well. The mixture was incubated in the dark at room temperature for 2 hours. The resulting blue color was measured using a microtiter plate reader. Gallic acid was used for calibration. The phenol content of the extract is expressed in terms of Gallic Acid Equivalent (GAE) concentration (mg/mL).
The results are shown in FIG. 5, and the plant polyphenol compound has strong free radical scavenging capacity and can prevent a series of chain reactions of free radicals, so that the plant polyphenol compound is a very efficient antioxidant. Therefore, the total content of the phenolic compounds in different extraction methods is measured by a colorimetric Folin-Ciocalteu method, and the more the antioxidant capacity of the mangosteen extract is found to be, the more the phenolic compounds are found to be. Therefore, the total polyphenol in the mangosteen extract acts as an effective component group to play an antioxidant effect.
Example 9 oral acute toxicity test of mangosteen extract
Based on the pre-test results, the contamination dose for the formal test was determined to be 5000mg/kg body weight. 30 rats subjected to formal tests are randomly divided into 3 groups, 10 rats (male and female halves) are taken from each group, 1 group is taken each time for test, and the test is repeated for 3 times; the test subjects were formulated with corn oil to be used as a solution for oral contamination of SD rats and ICR mice, and general manifestations, toxic symptoms and death of the rats were observed after contamination (general observation for 7 days after contamination of the formal test animals, if animals still died after contamination for 4 days, 14 days were required, if necessary, 28 days could be prolonged to be observed, and the died rats were examined by dissection and recorded.
The oral toxicity test results show that: the symptoms were absent after 1 hour of administration of the test substance, and no death was observed after 72 hours, and no abnormal changes were observed in tissues and organs of rats/mice examined by general dissection at the end of the test. In addition, mangosteen extract LD for oral acute toxicity test of SD rats 50 LD of mangosteen extract to ICR mice oral acute toxicity test greater than 5000mg/kg body weight 50 More than 5000mg/k g body weight, showing that the mangosteen extract is a low-toxicity or practically non-toxic plant extract.
The present invention is described in detail above. It will be apparent to those skilled in the art that the present invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with respect to specific embodiments, it will be appreciated that the invention may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.
Claims (8)
1. The preparation method of the mangosteen extract comprises the following steps: adding any one of the extraction solvents 1) -3) into mangosteen pericarp powder, heating and reflux-extracting, filtering, mixing filtrates, concentrating under reduced pressure until no alcohol smell exists, and lyophilizing to obtain mangosteen extract;
1) An aqueous solution of ethanol is used for preparing the aqueous solution,
2) The heating reflux extraction is carried out in two steps, wherein the extraction solvent used in the first step is 0.5-1 per mill formic acid aqueous solution, and the extraction solvent used in the second step is ethanol aqueous solution;
3) A mixed solvent of ethanol, ethyl acetate and water.
2. The method of manufacturing according to claim 1, characterized in that: 1) In the ethanol water solution, the volume concentration of ethanol is 50-90%;
2) In the second step, the volume concentration of the ethanol in the ethanol water solution is 50-90%;
3) In the mixed solvent of ethanol, ethyl acetate and water, the volume concentration of the ethanol is 45-90%, the volume concentration of the ethyl acetate is 0-50%, and the balance is water.
3. The preparation method according to claim 1 or 2, characterized in that: the ratio of the mangosteen pericarp powder to the extraction solvent is 1g:8 to 15L of the total weight of the product,
the temperature of the reflux is 100-140 ℃ and the time is 0.5-2.5 hours;
the heating reflux extraction is carried out for a plurality of times, specifically for 1 to 5 times;
the conditions of the decompression concentration are 40-70 ℃ and 0.07-0.08 MPa below zero.
4. A mangosteen extract prepared by the method of any one of claims 1-3, having an infrared spectrum at 3400cm -1 Characteristic absorption peak with hydroxyl group at about 3000-2850cm -1 The region has C-H telescopic vibration absorption peak of 1600cm -1 About 1300-1000cm with characteristic absorption peak of benzene ring -1 Has C-O telescopic vibration absorption peak;
the mangosteen extract mainly comprises alpha-mangostin and total polyphenol.
5. The use of the mangosteen extract of claim 4 for preparing an antibacterial, anti-inflammatory, antioxidant, growth-promoting related veterinary drug or feed additive in livestock and poultry farming.
6. The use according to claim 5, characterized in that:
the antibacterial is gram-positive,
the gram positive bacteria include sensitive staphylococcus aureus, methicillin-resistant staphylococcus aureus, vancomycin-resistant enterococci and clostridium perfringens.
7. The use according to claim 5, characterized in that:
the livestock and poultry comprise pigs, chickens and cattle.
8. The use according to claim 5, characterized in that:
the veterinary medicine or the medicine feed additive is administered orally or in a feed mixing and feeding mode;
the mangosteen extract can be used as veterinary medicine or medicine feed additive in the form of liquid preparation, powder or premix.
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| CN114452281A (en) * | 2022-02-23 | 2022-05-10 | 中国农业大学 | Application of mangosteen extract alpha-mangostin in preparation of medicine for preventing and treating clostridium perfringens infection of livestock and poultry |
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| CN114452281A (en) * | 2022-02-23 | 2022-05-10 | 中国农业大学 | Application of mangosteen extract alpha-mangostin in preparation of medicine for preventing and treating clostridium perfringens infection of livestock and poultry |
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| 陈文良;张孝友;陆原;: "热带植物山竹在美容护肤领域应用的研究进展", 农产品加工(学刊), no. 11, pages 77 - 79 * |
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