CN116251134B - Momordica grosvenori root blood sugar reducing composition, application and preparation method - Google Patents
Momordica grosvenori root blood sugar reducing composition, application and preparation method Download PDFInfo
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- CN116251134B CN116251134B CN202211580673.XA CN202211580673A CN116251134B CN 116251134 B CN116251134 B CN 116251134B CN 202211580673 A CN202211580673 A CN 202211580673A CN 116251134 B CN116251134 B CN 116251134B
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- momordica grosvenori
- grosvenori root
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Abstract
The invention relates to the technical field of traditional Chinese medicine extraction and preparation, in particular to a momordica grosvenori root hypoglycemic composition, application and a preparation method. The method comprises the steps of carrying out ultrasonic extraction on the momordica grosvenori roots, then carrying out adsorption and gradient elution on macroporous resin, concentrating and drying to obtain the momordica grosvenori root hypoglycemic composition. The Momordica grosvenori root hypoglycemic composition prepared by the method can obviously inhibit alpha-glucosidase, has IC 50 lower than acarbose, can obviously reduce the blood sugar level of db/db diabetic mice, increase insulin sensitivity and obviously reduce the expression of inflammatory factors such as IL-1 beta, IL-6, TNF-alpha and the like in serum of the diabetic mice at the dosage of 100mg/kg bw; has obvious improving effect on diabetes mice, and the blood sugar reducing mechanism can improve the insulin resistance of T2DM mice by inhibiting the inflammatory reaction of organisms. The preparation method has the advantages of simple process, rapidness, high efficiency, high purity, large-scale production and wide market prospect.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicine extraction and preparation, in particular to a momordica grosvenori root hypoglycemic composition, application and a preparation method.
Background
Recent survey reports from the international diabetes union (International Diabetes Federation, IDF) show that the number of people suffering from diabetes worldwide in 2017 is up to 4.25 million, and the number of people suffering from diabetes worldwide in 2045 is expected to increase to 6.29 million, and the number of people dying from diabetes-related diseases per year is up to 500 ten thousand. Diabetes is an important disease threatening the life health of people in China, and has great significance in actively and effectively preventing the progress of diabetes and related diseases. The natural medicine has the characteristics of low toxicity, safe use and obvious treatment and health care functions in controlling diabetes and complications, and can avoid adverse effects of common hypoglycemic medicines on diabetics in the treatment process to a great extent.
Momordica grosvenori [ Siraitia grosvenorii (Swingle) C.Jeffrey ] is a medicinal and edible plant in Guangxi Guilin road. The grosvenor momordica fruit industry is an important prop industry in Guilin areas, and the yield reaches billions. Momordica grosvenori has been used clinically for treating diabetes, pulmonary tuberculosis, acute and chronic tracheitis, etc. for over 300 years in folk medicinal history. Mogrosides, namely cucurbitane triterpene saponin, are main active ingredients of fructus Siraitiae Grosvenorii, and have effects of improving islet beta cell function, improving insulin resistance, and inhibiting glucose absorption in intestinal tract.
In recent years, the grosvenor momordica fruit industry is rapidly expanding, and the planting area of the grosvenor momordica fruit is developed from 12 ten thousand acres in 2016 to 21 ten thousand acres in 2021. At present, the cultivation mode of the momordica grosvenori is one year, the root tubers of the momordica grosvenori are dug every year, and along with the increase of the planting area, more and more momordica grosvenori root resources are generated. According to the records of Chinese herbal medicine and other documents, the Siraitia grosvenorii root has the functions of promoting diuresis and stopping diarrhea; relaxing tendons; major diarrhea; fat tongue; the function of the meningitis sequelae is a marked medicinal material for treating hepatitis and furuncle in Guilin areas. The main component of the momordica grosvenori root is cucurbitane triterpene, but is 19-bit carbon-loss triterpene or cucurbitacin and glycosides with novel structure, and the main component is different from the components of fruits. However, the research on the roots of momordica grosvenori is very limited, especially the research on pharmacological activity, so that the roots of momordica grosvenori are directly and usually discarded, and great resource waste is caused.
In the prior study on the Momordica grosvenori root, the related study is mainly focused on the separation and identification of chemical components of the Momordica grosvenori root, wang Xuefen, stoneware, chen Dihua and the like are sequentially separated from the Momordica grosvenori root to obtain the cucurbitane tetracyclic triterpenic acid (methyl-valerate and SIRAITIC F) without carbon, li Dianpeng and the like are separated from the Momordica grosvenori root to obtain two saponin components of the mogroside II and the mogroside II. However, the activity research of the related compounds is not reported in the paper.
None of the prior art relates to optimizing the extraction and preparation process of the momordica grosvenori root hypoglycemic component, nor does it disclose a method for rapidly identifying active ingredients therein.
Disclosure of Invention
The invention aims to solve the technical problem of providing a momordica grosvenori root hypoglycemic composition, application and a preparation method thereof.
The technical scheme for solving the technical problems is as follows:
The invention provides a preparation method of a momordica grosvenori root hypoglycemic composition, which comprises the following steps:
S1, ultrasonic extraction: obtaining momordica grosvenori root tuber powder, mixing the root tuber powder with a methanol aqueous solution, performing ultrasonic extraction and filtration, and removing part of alcohol solution in filtrate to obtain momordica grosvenori root extract;
In the methanol aqueous solution, the percentage of methanol is 40% -60% of the total volume of the methanol aqueous solution; the ratio of the volume of the methanol aqueous solution to the mass of the momordica grosvenori root tuber powder is 30:1-50:1 mL/g;
S2, adsorption and elution: adsorbing and gradient eluting the Momordica grosvenori root extract by using macroporous resin eluting column to obtain eluent;
S3, concentrating and drying: and concentrating and drying the eluent in sequence to obtain the momordica grosvenori root hypoglycemic composition.
In step S1, the ultrasonic extraction frequency is 10-50 kHz, the temperature is 30-50 ℃, and the ultrasonic extraction time is 30-60 min.
Further, in the step S2, the ratio of the weight of the macroporous resin elution column to the weight of the momordica grosvenori root tuber powder in the step S1 is 1:5-1:10.
Further, in the step S2, the eluent used in the gradient elution is a methanol solution; the gradient elution procedure is that methanol solutions with the volume percentages of 20%, 40% and 70% are respectively adopted for elution, and eluent after 70% methanol solution elution is collected; the volume of the methanol solution used for each elution is 2-4 times of the volume of the macroporous resin column.
Further, the macroporous resin in the macroporous adsorption resin elution column is one of D101 macroporous adsorption resin, HPD-100 macroporous adsorption resin and HP-20 macroporous resin.
Further, in the step S3, the eluent is sequentially subjected to reduced pressure concentration, spray drying, and freeze drying.
Further, spray drying is carried out by adopting a spray drying device; the temperature of the air inlet of the spray drying device is 140-165 ℃ and the temperature of the air outlet is 70-80 ℃.
Further, the temperature of the freeze drying is-50 to-40 ℃.
The invention also provides a momordica grosvenori root hypoglycemic composition obtained by the preparation method, and active ingredients in the composition comprise the following compounds: (-) -Larix Gmelini, 3,4' -dimethoxy-4,9,9 ' -trihydroxy-phenylpropanolignan-7 ' -ene, 23, 24-dihydrocucurbitacin F, mogroside III, mogroside II, mogroside IIb, mogroside II, mogroside IV, mogroside II, mogroside A II, mogroside II, 23, 24-dihydrocucurbitacin F-25-acetate, mogroside II, cucurbitacin B, 23, 24-dihydrocucurbitacin B, and dihydroisopcucurbitacin B-25 acetate.
The invention also provides application of the momordica grosvenori root hypoglycemic composition in preparation of hypoglycemic preparations.
The invention has the beneficial effects that:
(1) The preparation method of the momordica grosvenori root hypoglycemic composition adopts an ultrasonic extraction method, adopts 40-60% methanol aqueous solution, and can elute the hypoglycemic composition in the momordica grosvenori root tuber powder in combination with specific adsorption and elution steps, and the hypoglycemic composition prepared by the method has various active ingredients and good hypoglycemic effect;
(2) The momordica grosvenori root hypoglycemic composition can remarkably inhibit alpha-glucosidase, has an IC 50 lower than that of acarbose which is a commercially available hypoglycemic agent, can remarkably reduce the blood sugar level of db/db diabetic mice, increase insulin sensitivity and remarkably reduce the expression of inflammatory factors such as IL-1 beta, IL-6, TNF-alpha and the like in serum of the diabetic mice at a dosage of 100mg/kg bw;
(3) The momordica grosvenori root hypoglycemic composition has obvious improvement effect on diabetic mice, and the hypoglycemic mechanism of the composition improves the insulin resistance of T2DM mice by inhibiting inflammatory reaction of organisms;
(4) The preparation method of the momordica grosvenori root hypoglycemic composition has the advantages of simple process, rapidness, high efficiency, mass production and wide market prospect.
Drawings
FIG. 1 is a chromatogram of example 1, showing a method for preparing a Momordica grosvenori root hypoglycemic composition of the present invention; FIG. 1A shows an HPLC chromatogram of a Momordica grosvenori root hypoglycemic composition; FIG. 1B is an HPLC chromatogram of a Momordica grosvenori root hypoglycemic composition and an active alpha-glucosidase composition (a) and an inactive alpha-glucosidase composition (B);
FIG. 2 is a graph showing the comparison of the inhibition of alpha-glucosidase by different concentrations of acarbose, crude extract of Siraitia grosvenorii root, 40% methanol elution fraction and Siraitia grosvenorii root hypoglycemic composition according to the present invention;
FIG. 3 shows concentration levels of three inflammatory factors in the serum and tissue of each group of mice at the end of the experiment in example 2, which is a preparation method of the Momordica grosvenori root hypoglycemic composition of the present invention; FIG. 3A shows the concentration level of TNF- α, FIG. 3B shows the concentration level of IL-1β, and FIG. 3C shows the concentration level of IL-6;
FIG. 4 is a graph showing HPLC chromatograms of different extracts in comparative example 1, showing a method for preparing the Siraitia grosvenorii root hypoglycemic composition according to the present invention; fig. 4a is a chromatogram using water as an extraction solution, fig. 4B is a chromatogram using 50% methanol as an extraction solution, fig. 4C is a chromatogram using 50% ethanol as an extraction solution, fig. 4D is a chromatogram using 100% methanol as an extraction solution, and fig. 4E is a chromatogram using 100% ethanol as an extraction solution;
FIG. 5 is a HPLC liquid chromatogram of comparative example 2 using different extraction methods for preparing the Momordica grosvenori root hypoglycemic composition of the present invention; FIG. 5A shows a chromatogram of an ultrasonic extraction method, and FIG. 5B shows a chromatogram of an extraction method.
Detailed Description
The principles and features of the present invention are described below with reference to the drawings, the examples are illustrated for the purpose of illustrating the invention and are not to be construed as limiting the scope of the invention.
The preparation method of the momordica grosvenori root hypoglycemic composition adopts an ultrasonic extraction method, adopts 40-60% methanol aqueous solution, and can elute the hypoglycemic composition in the momordica grosvenori root tuber powder by combining specific adsorption and elution steps.
The preparation method specifically comprises the following steps:
S1, ultrasonic extraction: obtaining momordica grosvenori root tuber powder, mixing the root tuber powder with a methanol aqueous solution, performing ultrasonic extraction and filtration, and removing part of the alcohol solution in the filtrate to obtain a momordica grosvenori root extract; in the methanol aqueous solution, the percentage of methanol is 40% -60% of the total volume of the methanol aqueous solution; the ratio of the volume of the aqueous methanol solution to the mass of the momordica grosvenori root tuber powder is 30:1-50:1 mL/g. The purpose of extraction with 40-60% aqueous methanol solution is that the solvent can effectively extract the target component.
Preferably, the volume percent of the aqueous methanol solution is 50%.
Preferably, the ultrasonic extraction frequency is 10-50 kHz, the temperature is 30-40 ℃, and the ultrasonic extraction time is 30-60 min. The purpose of ultrasonic extraction is to accelerate release, diffusion and dissolution of effective substances in cells by cavitation, mechanical effect, thermal effect and the like of ultrasonic waves, and the extraction efficiency is remarkably improved.
Preferably, the preparation method of the momordica grosvenori root tuber powder comprises the steps of cleaning soil after the momordica grosvenori root tuber is collected, cutting the obtained product into slices, drying the slices in the shade at room temperature, and crushing the dried product to obtain the momordica grosvenori root tuber powder.
Preferably, the volume of the alcohol in the momordica grosvenori root extracting solution accounts for 15-20% of the total volume of the mixed solution. The purpose of the methanol solvent with 15-20% in the extracting solution is to ensure that some components with low binding force with macroporous resin are not adsorbed, but components with high binding force are effectively combined on the resin, thereby improving the adsorption selectivity and the desorption efficiency.
S2, adsorption and elution: absorbing and gradient eluting the Momordica grosvenori root extract by using a macroporous resin eluting column to obtain an eluent;
Wherein the ratio of the weight of the macroporous resin elution column to the weight of the momordica grosvenori root tuber powder in the step S1 is 1:5-1:10, and the eluent adopted by gradient elution is methanol solution; the gradient elution procedure is that methanol solutions with the volume percentages of 20 percent, 40 percent and 70 percent are respectively adopted for elution, and eluent after 70 percent of methanol solution elution is collected; the volume of the methanol solution used for each elution is 2-4 times of the volume of the macroporous resin column.
Preferably, the macroporous adsorbent resin is one of D101 macroporous adsorbent resin, HPD-100 macroporous adsorbent resin and HP-20 macroporous adsorbent resin.
S3, concentrating and drying: concentrating the eluent under reduced pressure until the dry matter content is 40-60%, and drying to obtain the Momordica grosvenori root hypoglycemic composition.
Preferably, the specific method of drying may be spray drying or freeze drying, or both may be used. The spray drying method is carried out by adopting a spray drying device, the temperature of an air inlet of the spray drying device is 140-165 ℃, and the temperature of an air outlet of the spray drying device is 70-80 ℃.
Preferably, the temperature of freeze-drying is-50 to-40 ℃.
The preparation method provided by the invention has the advantages of simple process, rapidness, high efficiency and high purity, and the composition can be produced in a large scale.
The momordica grosvenori root hypoglycemic composition comprises the following active ingredients: (-) -Larix Gmelini, 3,4' -dimethoxy-4,9,9 ' -trihydroxy-phenylpropanolignan-7 ' -ene, 23, 24-dihydrocucurbitacin F, mogroside III, mogroside II, mogroside IIb, mogroside II, mogroside IV, mogroside II, mogroside A II, mogroside II, 23, 24-dihydrocucurbitacin F-25-acetate, mogroside II, cucurbitacin B, 23, 24-dihydrocucurbitacin B, and dihydroisopcucurbitacin B-25 acetate.
Experiments prove that the momordica grosvenori root blood sugar reducing composition can remarkably inhibit alpha-glucosidase, has IC 50 lower than acarbose, can remarkably reduce the blood sugar level of db/db diabetic mice at the dosage of 100mg/kg bw, increases insulin sensitivity, and simultaneously remarkably reduces the expression of inflammatory factors such as IL-1 beta, IL-6, TNF-alpha and the like in serum of the diabetic mice. Thus, the composition has obvious improvement effect on diabetic mice, and the blood glucose reducing mechanism is probably to improve the insulin resistance of T2DM mice by inhibiting inflammatory reaction of organisms.
The momordica grosvenori root hypoglycemic composition can be applied to preparation of hypoglycemic preparations.
Specifically, the general preparation method can be used for preparing oral medicaments such as capsules, injection and the like containing the hypoglycemic composition.
The technical scheme of the invention is specifically described below by specific examples and comparative examples:
Example 1 identification of hypoglycemic active Components in Momordica grosvenori Swingle root hypoglycemic Components
The embodiment specifically identifies the hypoglycemic active ingredient in the momordica grosvenori root hypoglycemic component of the invention so as to verify the specific active ingredient type in the composition.
The authentication process of this embodiment includes the steps of:
(1) The Momordica grosvenori root hypoglycemic composition is combined with alpha-glucosidase.
Specifically, 100. Mu.L of Momordica grosvenori root hypoglycemic composition with concentration of 2.0mg/mL is mixed with 100. Mu.L of alpha-glucosidase PBS solution, wherein the alpha-glucosidase PBS solution is 1U/mL, and pH is 6.8. After mixing, the mixture was incubated at 37℃for 30min. Meanwhile, 100. Mu.L of alpha-glucosidase was boiled in boiling water for 10min to inactivate and then mixed with 100. Mu.L of Momordica grosvenori root hypoglycemic composition having a concentration of 2.0mg/mL, and cultured at 37℃for 30min as a negative control.
(2) Ultrafiltration membrane interception and elution: the solution mixture of step (1) was transferred to a 10KD ultrafiltration centrifuge tube and centrifuged at 10000rpm for 10min at room temperature. The filter residue was washed three times with 200. Mu.L of PBS solution (pH 6.8 or pH7.4,25 ℃) to remove potentially non-specifically bound or free compounds. Then 200. Mu.L of 90% (v/v) acetonitrile was added for dissolution for 10min, the above procedure was repeated twice by centrifugation at 10000rpm for 10min at room temperature, and the 90% acetonitrile solution was collected, concentrated and dried to obtain a fraction specifically binding to alpha-glucosidase. In addition, alpha-glucosidase was heat inactivated in a water bath at 100℃for 10min as a negative control.
(3) HPLC analysis identified components that specifically bind to α -glucosidase: the component specifically binding to α -glucosidase was obtained in step (2) by dissolving with 50 μl of 90% acetonitrile, and the monomer compound was analyzed by HPLC. HPLC analysis was performed using an Inertsil ODS-3 (250X 4.6mm,5 μm) analytical column, column temperature at 35℃, flow rate 1mL/min, sample volume 10. Mu.L, detection wavelength 210nm; water and acetonitrile are used as mobile phases, and gradient elution is carried out: 0-40min,20% -50% acetonitrile. Compared with the HPLC chromatogram of the Momordica grosvenori root hypoglycemic composition and the inactive alpha-glucosidase binding component, the HPLC chromatogram peak area of the Momordica grosvenori root hypoglycemic composition and the active alpha-glucosidase binding component is increased, the chromatogram peak is considered to be the active chromatogram peak, and the compound corresponding to the chromatogram peak is confirmed by comparing with the HPLC retention time and the ultraviolet absorption characteristic of the standard substance.
As shown in FIG. 1, 17 components having an alpha-glucosidase inhibitory activity were identified in the Siraitia grosvenorii root hypoglycemic composition by the above identification method. The structures of 16 compounds have been identified, which are (-) -lariciresinol, 3,4' -dimethoxy-4,9,9 ' -trihydroxy-phenylpropanolignan-7 ' -ene, 23, 24-dihydrocucurbitacin F, mogroside III, mogroside II, mogroside IIb, mogroside II, mogroside IV, mogroside II,23, 24-dihydrocucurbitacin F-25-acetate, mogroside II, cucurbitacin B,23, 24-dihydroisoparaffinin B-25 acetate, respectively.
As shown in figure 2, the crude extract of the Momordica grosvenori root, 40% methanol elution part and the composition for reducing blood sugar of the Momordica grosvenori root have positive correlation with the inhibition rate of alpha-glucosidase within a certain mass concentration range. When the mass concentration is the same, the inhibition rate of the alpha-glucosidase is greatly different, and the strength sequence is as follows: momordica grosvenori root hypoglycemic composition > acarbose >40% methanol elution part > Momordica grosvenori root crude extract. The result shows that the alpha-glucosidase inhibitory activity of the grosvenor momordica root crude body is improved through macroporous resin, and the obtained 40% methanol elution part and the grosvenor momordica fruit hypoglycemic composition (70% methanol elution part) are eluted by using 20%, 40% and 70% methanol, wherein the activity inhibition effect of the grosvenor momordica root hypoglycemic composition (IC 50 = 0.628 +/-0.027) sample on the alpha-glucosidase is superior to that of positive control acarbose (IC 50 =0.855+/-0.028), and the part of the sample has better alpha-glucosidase inhibitory capacity, so that the part is considered to be the grosvenor momordica root hypoglycemic component.
Example 2 alpha-glucosidase inhibitory Activity experiment of Momordica grosvenori Swingle root hypoglycemic composition
This example performed an experiment of alpha-glucosidase inhibitory activity on the momordica grosvenori root hypoglycemic composition according to the present invention.
In this example, PNPG method was used for activity study, 50. Mu.L of phosphate buffer (50 mM concentration, pH 6.8) was added to a 96-well plate, then 20. Mu.L of the sample solution to be tested and 20. Mu.L of enzyme solution (1U/mL) were added respectively, mixed by shaking, placed in a 37℃incubator for preheating for 5min, then 20. Mu.L of PNPG (1 mM) was added, reacted in a 37℃incubator for 30min, and finally 50. Mu.L of anhydrous sodium carbonate solution (0.2M) was added to terminate the reaction, and the absorbance at a wavelength of 405nm was measured. Distilled water is used to replace the sample to make blank, and other steps and reagents are the same as above. The enzyme activity inhibition ratio at each of the series concentrations was calculated, and from this, the IC 50 value was calculated.
Enzyme activity inhibition rate= (A Blank space -(A Sample of -A Background )/A Blank space multiplied by 100%, wherein, A blank is the light absorption value after the reaction of the sample to be detected is not added, A sample is the light absorption value after the reaction of the sample to be detected, enzyme and substrate inhibition is added, and A background is the light absorption value after the reaction of the sample to be detected is only added.
In the hypoglycemic composition, the alpha-glucosidase inhibitory activities of 16 compounds are shown in table 1, acarbose is used as a positive control, and all compounds 1-16 show a certain inhibitory activity of alpha-glucosidase and have an inhibitory rate of 19.73% -70.67% at a mass concentration of 1mg/mL, wherein the activity of the compounds 1,2 and 14 is superior to that of the acarbose sold in the market.
TABLE 1 inhibition Activity of alpha-glucosidase
Note that: the compound concentration measured in a was 1mg/mL, n.d.: IC 50 measurement was not performed due to insufficient amount of monomer compound
Example 3 experiments on hypoglycemic Activity of Momordica grosvenori Swingle root hypoglycemic composition in mice
In this example, wild-type C57BLKS (db/dm) mice were used as a blank, db/db mice were randomly divided into a model group and a dosing group, and Momordica grosvenori root hypoglycemic composition (100 mg/kg) was administered by intragastric administration to the dosing group, and distilled water was administered to the blank control group and the model group, respectively, for 56 days. The body weight of the mice is measured at the end of the experiment, and the biochemical indexes of blood and liver are detected, and the results in table 2 and fig. 3 show that the blood glucose lowering composition of the fructus momordicae has the effect of reducing the blood glucose of the diabetic mice, and is related to improving the inflammatory response and insulin resistance of the mice, as compared with the untreated group, due to the fact that the blood glucose lowering composition of the fructus momordicae has the obvious lower levels of TNF-alpha, IL-1 beta and IL-6 in the blood of the db/db mice, such as FBG (fiber), fasting insulin index (Fins) and insulin resistance level (HOMA-IR).
TABLE 2 comparison of FBG, fins, HOMA-IR of mice of each group at the end of the experiment
FBG, fasting blood glucose; fins, fasting insulin; HOMA-IR, calculating insulin resistance index by using steady state model method; a Compared with the blank control group; b P < 0.05 compared to untreated group.
Comparative example 1 comparison of different extraction solvents
Taking 2.0g of 5 parts of momordica grosvenori root tuber powder, respectively adding water, 50% methanol, 50% ethanol, 100% methanol and 100% ethanol, shaking uniformly, performing ultrasonic extraction for 1h, and measuring the extract by utilizing HPLC.
The chromatogram of the above measurement is shown in FIG. 4. As can be seen from the comparison analysis of the chromatograms, the extraction effect is best by 50% methanol solution, and the peak area of 9 common peaks is maximum; the ethanol solution has poor extraction effect, the minimum peak is obtained, and the peak area of 9 common peaks in 100% ethanol solvent extraction is relatively small, so that ethanol is removed as an extraction solvent; the extraction is carried out by taking water as a solvent, the main component saponin compound in the momordica grosvenori tubers cannot be completely extracted, and finally 50% methanol is determined to be used as an extraction solvent.
Comparative example 2 ultrasonic extraction and extraction method comparison
(1) Ultrasonic extraction: 2.0g of momordica grosvenori root tuber powder is taken, precisely weighed, 50ml of 50% methanol is added, the mixture is uniformly shaken, ultrasonic treatment is carried out for 1h at room temperature, and the supernatant is taken for HPLC analysis;
(2) Soaking and extracting: 2.0g of momordica grosvenori root tuber powder is taken, precisely weighed, 50ml of 50% methanol is added, the mixture is shaken uniformly and is subjected to cold soaking, the mixture is left at room temperature overnight for about 12 hours, and the supernatant is taken for HPLC analysis.
The HPLC analysis results are shown in FIG. 5. As can be seen from the chromatogram, the sample extracted for 1 hour by ultrasonic extraction has obvious chromatographic peaks under the same solvent, while the sample extracted by soaking for 12 hours has almost no peaks except the chromatographic peaks before 5 minutes, which fully indicates that the ultrasonic extraction efficiency is higher.
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.
Claims (7)
1. The preparation method of the momordica grosvenori root hypoglycemic composition is characterized by comprising the following steps of:
S1, ultrasonic extraction: obtaining momordica grosvenori root tuber powder, mixing the momordica grosvenori root tuber powder with a methanol aqueous solution, performing ultrasonic extraction and filtration, and removing part of alcohol solution in filtrate to obtain momordica grosvenori root extract;
In the methanol aqueous solution, the percentage of methanol is 40% -60% of the total volume of the methanol aqueous solution; the ratio of the volume of the methanol aqueous solution to the mass of the momordica grosvenori root tuber powder is 30:1-50:1 mL/g;
S2, adsorption and elution: adsorbing and gradient eluting the Momordica grosvenori root extract by using macroporous adsorption resin chromatographic column to obtain eluent;
S3, concentrating and drying: concentrating and drying the eluent in sequence to obtain the momordica grosvenori root hypoglycemic composition;
In the step S2, the ratio of the weight of the macroporous resin to the weight of the momordica grosvenori root tuber powder in the step S1 is 1:5-1:10;
in the step S2, the eluent adopted in the gradient elution is methanol solution; the gradient elution procedure is that methanol solutions with the volume percentages of 20%, 40% and 70% are respectively adopted for elution, and eluent after 70% methanol solution elution is collected; the volume of the methanol solution used for each elution is 2-4 times of the volume of the macroporous resin column;
The active ingredients in the composition include the following compounds: (-) -lariciresinol, 3,4' -dimethoxy-4,9,9 ' -trihydroxy-phenylpropanolignan-7 ' -ene, 23, 24-dihydrocucurbitacin F, mogroside III, mogroside II, mogroside IIb, mogroside II, mogroside IV, mogroside II, mogroside A II, mogroside II, 23, 24-dihydrocucurbitacin F-25-acetate, mogroside II, cucurbitacin B, 23, 24-dihydrocucurbitacin B, dihydroisopcucurbitacin B-25 acetate;
the volume of the alcohol in the momordica grosvenori root extracting solution accounts for 15-20% of the total volume of the mixed solution.
2. The method for preparing the momordica grosvenori root hypoglycemic composition according to claim 1, wherein in the step S1, the ultrasonic extraction frequency is 10-50 kHz, the temperature is 30-50 ℃, and the ultrasonic extraction time is 30-60 min.
3. The method for preparing the momordica grosvenori root hypoglycemic composition according to any one of claims 1-2, wherein the macroporous resin in the macroporous adsorption resin chromatographic column is one of D101 type macroporous adsorption resin, HPD-100 type macroporous adsorption resin and HP-20 type macroporous resin.
4. The method for preparing a momordica grosvenori root hypoglycemic composition according to any one of claims 1-2, wherein in the step S3, the eluent is subjected to reduced pressure concentration, spray drying and freeze drying in sequence.
5. The method for preparing the momordica grosvenori root hypoglycemic composition according to claim 4, wherein the step of spray drying is performed by a spray drying device; the air inlet temperature of the spray drying device is 140-165 ℃, and the air outlet temperature is 70-80 ℃.
6. The method for preparing the momordica grosvenori root hypoglycemic composition according to claim 4, wherein the freeze-drying temperature is-50 to-40 ℃.
7. Use of the momordica grosvenori root hypoglycemic composition according to any one of claims 1-6 in the preparation of hypoglycemic preparations.
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| 罗汉果根的生物活性及化学成分研究;扈芷怡等;《吉林农业大学学报》;第1-10页 * |
| 高效液相色谱法测定罗汉果根中两个降三萜配糖体;卢凤来等;《广西植物》;第30卷(第6期);第891-894页 * |
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