CN113398157A - Method for continuously extracting and separating multiple natural active ingredients from momordica grosvenori flower - Google Patents
Method for continuously extracting and separating multiple natural active ingredients from momordica grosvenori flower Download PDFInfo
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Abstract
The method for continuously extracting and separating a plurality of natural active ingredients from the momordica grosvenori flower comprises the following steps: (1) extracting volatile oil: crushing dried momordica grosvenori flowers, adding an organic solvent for reflux extraction to obtain an organic solvent extracting solution containing volatile oil and extraction residues containing total flavonoids and total saponins; (2) purifying the volatile oil; (3) extracting total flavonoids and total saponins: refluxing and extracting the extraction slag; (4) enrichment of total flavonoids and total saponins: concentrating the alcoholic extractive solution to obtain concentrated solution containing total flavonoids and total saponins; (5) and (3) separation of total saponins: decolorizing the concentrated solution with macroporous anion exchange resin column to obtain total saponin product; (6) and (3) separating total flavonoids: eluting the anion exchange resin column after the total saponin is separated with alkali liquor to obtain the total flavone product. The invention can continuously extract and separate various natural active ingredients, and has the advantages of high content of the obtained product, high yield, low requirement on equipment and low production cost.
Description
Technical Field
The invention relates to a method for extracting and separating natural active ingredients from momordica grosvenori flowers, in particular to a method for continuously extracting and separating a plurality of natural active ingredients from momordica grosvenori flowers.
Background
Fructus Siraitiae Grosvenorii flower has effects of clearing heat and detoxicating, eliminating phlegm and relieving cough, nourishing sound and moistening lung, and can be used for treating diseases such as halitosis, stomatitis, pharyngitis, tonsillitis, and acne. After artificial pollination, the momordica grosvenori flowers, especially male flowers, are usually treated as waste, which not only wastes natural resources, but also causes environmental pollution to a certain extent. Therefore, if a plurality of natural active ingredients with pharmacological effects can be extracted and separated from the momordica grosvenori flowers, the wastes are changed into valuable substances, and the method has important practical significance for protecting the environment, promoting the comprehensive utilization of momordica grosvenori resources and promoting the income increase of planting bases and momordica grosvenori processing enterprises.
The momordica grosvenori flower contains natural active ingredients such as volatile oil, flavonoid compounds, saponin compounds and the like.
The momordica grosvenori flower volatile oil mainly contains terpenes such as alpha-pinene, beta-pinene, camphene, myrcene, limonene and the like, aromatic hydrocarbons, aromatic esters and the like. The volatile oil components can eliminate inflammation of trachea and lung, relieve cough and reduce sputum, and has obvious effect of relieving adverse symptoms such as cough, excessive phlegm, dyspnea, and the like caused by cold and tracheitis. The flavonoids compounds in the flowers of momordica grosvenori mainly use kaempferol as aglycone, and the compounds have the medicinal health-care functions of resisting oxidation and aging, treating cardiovascular and cerebrovascular diseases, reducing blood sugar, resisting cancer and the like. In addition, although mogrosides (a natural high intensity sweetener) unique to the fruit of momordica grosvenori is not contained in the flowers of momordica grosvenori, other types of saponins are still contained therein. The mogroside has good bacteriostatic activity, and is a natural bacteriostatic agent.
So far, there have been few studies on extraction of effective ingredients from momordica grosvenori flowers.
CN113041279A at 29.06/29/2021 discloses a method for separating effective components from luo han guo flowers. The method comprises the following steps: s1, extracting the momordica grosvenori flower with water vapor to obtain wet flower and a first extracting solution; s2, extracting the wet flower with ethanol as a solvent at 25-65 ℃ to obtain a second extracting solution and first filter residue, concentrating the second extracting solution, and carrying out solid-liquid separation to obtain a third extracting solution and wet residue; s3, concentrating and standing the third extracting solution to obtain crystals and crystallization mother liquor; and S4, passing the crystallized mother liquor through a macroporous adsorption resin chromatographic column, and eluting with an acid solution, an alkali solution and an alcohol solution in sequence to respectively obtain an acid-eluted liquid, an alkali-eluted liquid and an alcohol-eluted liquid. However, the method is complex to operate, the content of active ingredients of the obtained product is low, the used solvent amount is large, and the production cost is high.
Disclosure of Invention
The technical problem to be solved by the invention is to overcome the defects in the prior art and provide a method for continuously extracting and separating a plurality of natural active ingredients from momordica grosvenori flowers, wherein the obtained product has high active ingredient content, high yield, low requirement on equipment and low production cost.
The technical scheme adopted for solving the technical problems is that the method for continuously extracting and separating a plurality of natural active ingredients from the flowers of momordica grosvenori comprises the following steps:
(1) extracting volatile oil: crushing dried momordica grosvenori flower, placing in an extraction tank, adding an organic solvent, performing reflux extraction, and filtering to obtain an organic solvent extract containing volatile oil and extraction residues containing total flavonoids and total saponins;
(2) and (3) volatile oil purification: adding a decoloring agent into the organic solvent extracting solution obtained in the step (1), stirring and decoloring, filtering, collecting filtrate, and concentrating the filtrate to obtain a momordica grosvenori flower volatile oil product;
(3) extracting total flavonoids and total saponins: distilling the organic solvent in the extraction residue obtained in the step (1) to be clean, then adding methanol or ethanol, carrying out reflux extraction, and filtering to obtain an alcohol extract;
(4) enrichment of total flavonoids and total saponins: concentrating the alcohol extract obtained in the step (3) under reduced pressure, adsorbing by using a macroporous adsorption resin column, eluting the macroporous adsorption resin column by using an eluent ethanol, collecting and concentrating the eluent to obtain a concentrated solution containing the total flavone and the total saponin;
(5) and (3) separation of total saponins: decoloring the concentrated solution containing the total flavonoids and the total saponins obtained in the step (4) by using a macroporous anion exchange resin column, collecting the effluent of the anion exchange resin column, concentrating under reduced pressure, and drying to obtain a total saponin product;
(6) and (3) separating total flavonoids: eluting the anion exchange resin column after the total saponins are separated in the step (5) by using alkali liquor to obtain alkali elution liquid; neutralizing the alkaline eluent with acid, desalting with nanofiltration membrane, collecting the nanofiltration membrane retentate, concentrating under reduced pressure, and drying to obtain total flavone product.
Preferably, in the step (1), the organic solvent is a mixed solvent of a low-polarity organic solvent and ethyl acetate.
Further, the low-polarity organic solvent is preferably diethyl ether, petroleum ether, hexane, cyclohexane, heptane, 6#Solvent oil and 120#One or more solvent oils.
Further, the volume ratio of the low-polarity organic solvent to the ethyl acetate is 5-10: 1, preferably 8-9: 1.
the purpose of adding ethyl acetate into the low-polarity organic solvent is to properly improve the polarity of the mixed organic solvent, so that the volatile oil in the momordica grosvenori flowers can be leached more fully.
Preferably, in the step (1), the amount of the organic solvent is 10-20 times (L/kg) of the weight of the raw material momordica grosvenori flower; the reflux extraction time is 2-4 hours.
The purpose of reflux extraction by using mixed organic solvent is to more fully extract the volatile oil in the momordica grosvenori flower. If a single organic solvent is used, or the dosage of the mixed organic solvent is too small or the reflux extraction time is too short, the purpose cannot be fully achieved; if the dosage of the mixed organic solvent is too much or the reflux extraction time is too long, the waste of materials and energy sources is caused.
Preferably, in the step (2), the decolorizing agent is one or more of activated carbon, activated clay, diatomite, alumina adsorbent resin and silica gel adsorbent resin; the preferable dosage of the decolorant is 0.1-2% of the weight of the raw material momordica grosvenori flower; the stirring and decoloring time is 1-4 hours, preferably 2-3 hours.
Stirring and decolorizing with decolorizing agent to remove pigment and impurities in the mixed organic solvent extractive solution to obtain high-quality flos Momordicae volatile oil.
Preferably, in the step (3), the volume fraction of the methanol or the ethanol is 50 to 90 percent, preferably 70 to 85 percent; the dosage of the methanol or the ethanol is 10-20 times (L/kg) of the weight of the raw material momordica grosvenori flower; the reflux extraction time is 2-4 hours.
The purpose of reflux extraction by using methanol or ethanol is to fully leach out the total flavone and the total saponin in the mixed organic solvent extraction residue. If the volume fraction of the methanol or the ethanol is too small, the dosage is too small or the reflux extraction time is too short, the aim can not be fully achieved; if the volume fraction of methanol or ethanol is too large, the dosage is too large or the reflux extraction time is too long, the waste of materials and energy sources is caused.
Preferably, the model of the macroporous adsorption resin is D101, AB-8, LX-38 or DM130, and the dosage (volume) of the macroporous adsorption resin is 0.5-1 time (L/kg) of the weight of the raw material momordica grosvenori flower.
Further, the height-diameter ratio of the macroporous adsorption resin column is preferably 4-6: 1; the flow rate of the upper column is 0.5-1.0 BV/h, preferably 0.55-0.8 BV/h.
The macroporous adsorption resin column is used for adsorbing the total flavonoids and the total saponins of the momordica grosvenori flower in the concentrated solution of the alcohol extract so as to achieve the aim of enrichment.
Preferably, in step (5), the macroporous anion exchange resin is D941, D945, LSA-700 or LSD 762.
Further, the dosage (volume) of the anion resin is 0.5-1 time (L/kg) of the weight of the raw material momordica grosvenori flower; the height-diameter ratio of the anion exchange resin column is 4-6: 1.
Further, the flow rate of the upper column is 0.5-1.0 BV/h; preferably 0.55-0.8 BV/h.
One of the purposes of using an anion exchange resin column is to remove impurities and decolor so as to obtain the grosvenor momordica flower total saponins with higher content; the second purpose is to adsorb the total flavonoids of the momordica grosvenori flowers on anion exchange resin to achieve the purpose of separating the total saponins and the total flavonoids. If the dosage of the anion exchange resin is too small, the height-diameter ratio of the anion exchange resin column is too small or the flow rate of the anion exchange resin column is too high, the purpose cannot be fully achieved; if the dosage of the anion exchange resin is too much, the height-diameter ratio of the anion exchange resin column is too large or the flow rate of the anion exchange resin column is too low, not only can the waste of materials and energy be caused, but also the difficulty and the treatment capacity of subsequent elution can be increased.
Preferably, in the step (6), the alkali liquor is an aqueous solution of sodium hydroxide or potassium hydroxide, the mass percentage concentration of the alkali liquor is 2% -5%, and the use amount of the alkali liquor is 2-3 BV.
The purpose of eluting the anion exchange resin column with an alkali solution is to elute the total flavonoids adsorbed on the anion exchange resin. If the mass percentage concentration of the alkali liquor is too low or the dosage of the alkali liquor is too small, the total flavone cannot be completely eluted, so that the yield of the total flavone is low; if the mass percentage concentration of the alkali liquor is too high or the dosage of the alkali liquor is too much, not only the waste of materials and energy sources is caused, but also the molecular structure of the flavonoid compound is possibly damaged, and the yield of the total flavone is also low.
Preferably, in the step (6), the acid is hydrochloric acid or sulfuric acid, and the mass percentage concentration of the acid is 5-15%. The purpose of using acid for neutralization is to convert the total flavone in ion state in the alkali liquor eluent into molecular state, and finally obtain the free total flavone.
Preferably, in the step (6), the molecular weight cut-off of the nanofiltration membrane is 100-500 Da, preferably 200-300 Da; the filtration pressure is preferably 0.3-0.6 MPa.
The purpose of using nanofiltration membrane for filtration is to remove a large amount of inorganic salts in the neutralized eluate, thereby increasing the content of total flavonoids in the product. If the molecular weight cut-off of the nanofiltration membrane is too low or the filtering pressure is too low, inorganic salt cannot sufficiently permeate the nanofiltration membrane, so that the desalting effect is poor, and the content of total flavonoids is low; if the trapped molecular weight of the nanofiltration membrane is too large or the filtering pressure is too large, part of the flavonoid compounds pass through the nanofiltration membrane and exist in the permeate of the nanofiltration membrane, so that the yield of the total flavonoids is low.
In the method of the invention, 1 BV =1 resin column volume.
The principle of the method of the invention is as follows: reflux-extracting flos Momordicae with organic solvent (preferably mixed solvent of low-polarity organic solvent and ethyl acetate) to obtain flos Momordicae volatile oil, rather than total saponins and total flavonoids; reflux-extracting the organic solvent extraction residue with lower alcohol (preferably methanol or ethanol) to extract total saponins and total flavonoids; the flavonoids compounds contain phenolic hydroxyl and can be ionized into an ionic form, so that the macroporous anion exchange resin can adsorb the flavonoids compounds (the grosvenor momordica flower total saponins do not contain phenolic hydroxyl and cannot be ionized, so that the grosvenor momordica flower total saponins cannot be adsorbed by the macroporous anion exchange resin), thereby realizing the separation of the grosvenor momordica flower total saponins and the grosvenor momordica flower total flavonoids; finally, the flavonoids adsorbed on the anion exchange resin can be eluted by the lye.
Compared with the prior art, the invention has the following beneficial effects:
(1) in the obtained momordica grosvenori flower volatile oil product, the content of the volatile oil can be as high as 93.95%, and the yield can be as high as 95.75%; in the grosvenor momordica flower total saponin product, the content of the total saponin can reach 91.77 percent, and the yield can reach 94.04 percent; the content of the total flavonoids in the momordica grosvenori flower total flavonoids product can reach 91.35 percent, and the yield can reach 94.48 percent;
(2) the method has the advantages of strong operability of the process, low cost and low requirement on equipment, realizes the high-efficiency comprehensive utilization of the momordica grosvenori resource, opens up a new way for increasing the income of momordica grosvenori processing enterprises, and is suitable for industrial production.
Detailed Description
The present invention is further illustrated by the following specific examples. It should be noted that the described embodiments illustrate only some of the embodiments of the invention, and should not be construed as limiting the scope of the claims. All other changes and modifications which can be made by one skilled in the art based on the embodiments of the present invention without inventive faculty are within the scope of the claims of the present application.
The dry petals of the momordica grosvenori used in the embodiment of the invention are purchased from organic momordica grosvenori planting bases of the Hunan Huacheng biological resource GmbH, wherein the mass percent contents of the volatile oil, the total saponin and the total flavone are respectively 6.09%, 1.92% and 5.51%; the macroporous adsorption resin and the anion exchange resin used in the embodiment of the invention are purchased from Xian lan Xiao science and technology New materials GmbH; the starting materials or chemicals used in the examples of the present invention are, unless otherwise specified, commercially available in a conventional manner.
Example 1
The embodiment comprises the following steps:
(1) extracting volatile oil: crushing 10kg of dried momordica grosvenori flower to the particle size of 2-5 mm, placing the crushed momordica grosvenori flower in an extraction tank, adding 200L of mixed organic solvent (petroleum ether: ethyl acetate = 8: 1, V/V), extracting for 2.5 hours under reflux, and filtering to obtain organic solvent extract containing volatile oil and extraction residues containing total flavonoids and total saponins;
(2) and (3) volatile oil purification: adding 0.1kg of silica gel adsorption resin into the organic solvent extract obtained in the step (1), stirring and decoloring for 3 hours, filtering, collecting filtrate, and concentrating the filtrate to obtain 0.63kg of a momordica grosvenori flower volatile oil product;
(3) extracting total flavonoids and total saponins: distilling the organic solvent in the extraction residue obtained in the step (1) to be clean, then adding 180L of 80% methanol solution by volume concentration, carrying out reflux extraction for 3 hours, and filtering to obtain an alcohol extract;
(4) enrichment of total flavonoids and total saponins: concentrating the alcohol extract obtained in the step (3) under reduced pressure, adsorbing by using a macroporous adsorption resin column (the model of the macroporous adsorption resin is D101, the dosage of the macroporous adsorption resin is 9L, the height-diameter ratio of the macroporous adsorption resin column is 5:1, and the flow rate of the macroporous adsorption resin column is 0.7 BV/h), eluting the macroporous adsorption resin column by using absolute ethyl alcohol, collecting and concentrating eluent, and obtaining concentrated solution containing the total flavonoids and the total saponins of the momordica grosvenori flower;
(5) and (3) separation of total saponins: decoloring the concentrated solution containing the total flavonoids and the total saponins obtained in the step (4) by using a macroporous anion exchange resin column (the model of the macroporous anion exchange resin is D941, the using amount of the anion resin is 9L of the weight of the raw material momordica grosvenori flower, the height-diameter ratio of the anion exchange resin column is 5:1, the flow rate of the anion exchange resin column is 0.6 BV/h), collecting the effluent of the anion exchange resin column, concentrating under reduced pressure, and drying to obtain 0.20kg of momordica grosvenori flower total saponins;
(6) and (3) separating total flavonoids: eluting the anion exchange resin column after the total saponins are separated in the step (5) by using 3BV of sodium hydroxide aqueous solution with the mass percent concentration of 3 percent to obtain alkaline elution liquid; neutralizing the alkaline eluent with hydrochloric acid with a mass percent concentration of 8%, desalting with a nanofiltration membrane (the cut-off molecular weight of the nanofiltration membrane is 100Da, and the filtering pressure is 0.6 Mpa), collecting the nanofiltration membrane cut-off liquid, concentrating the nanofiltration membrane cut-off liquid under reduced pressure, and drying to obtain 0.57kg of the total flavonoids of the momordica grosvenori flowers.
According to the determination of the method specified in the Chinese pharmacopoeia, the content of the volatile oil in the momordica grosvenori flower volatile oil product obtained in the embodiment is 92.56%, and the yield is 95.75%; through ultraviolet spectrophotometry, the content of the total saponins in the momordica grosvenori flower total saponins product obtained in the embodiment is 89.43%, and the yield is 93.16%; the content of the total flavonoids in the momordica grosvenori flower total flavonoids product obtained in the embodiment is 90.19% and the yield is 93.30% through ultraviolet spectrophotometry.
Example 2
The embodiment comprises the following steps:
(1) extracting volatile oil: crushing 10kg of dried momordica grosvenori flower to the particle size of 2-5 mm, placing the crushed momordica grosvenori flower in an extraction tank, adding 180L of mixed organic solvent (6 # solvent oil: ethyl acetate = 10: 1, V/V), extracting for 3 hours under reflux, and filtering to obtain organic solvent extract containing volatile oil and extraction residues containing total flavonoids and total saponins;
(2) and (3) volatile oil purification: adding 0.15kg of diatomite into the organic solvent extract obtained in the step (1), stirring and decoloring for 2 hours, filtering, collecting filtrate, and concentrating the filtrate to obtain 0.62kg of a momordica grosvenori flower volatile oil product;
(3) extracting total flavonoids and total saponins: distilling the organic solvent in the extraction residue obtained in the step (1) to be clean, then adding 160L of 85% ethanol solution with volume concentration, carrying out reflux extraction for 3.5 hours, and filtering to obtain an ethanol extract;
(4) enrichment of total flavonoids and total saponins: concentrating the alcohol extract obtained in the step (3) under reduced pressure, adsorbing by using a macroporous adsorption resin column (the model of the macroporous adsorption resin is AB-8, the using amount of the macroporous adsorption resin is 8L, the height-diameter ratio of the macroporous adsorption resin column is 6:1, and the flow rate of the macroporous adsorption resin column is 0.8 BV/h), eluting the macroporous adsorption resin column by using absolute ethyl alcohol, collecting, and concentrating the eluent to obtain a concentrate containing the total flavonoids and the total saponins of the momordica grosvenori flower;
(5) and (3) separation of total saponins: decolorizing the concentrated solution containing the total flavonoids and the total saponins obtained in the step (4) by using a macroporous anion exchange resin column (the model of the macroporous anion exchange resin is D945, the dosage of the anion resin is 8L of the weight of the raw material momordica grosvenori flower, the height-diameter ratio of the anion exchange resin column is 6:1, the flow rate of the anion exchange resin column is 0.8 BV/h), collecting the effluent of the anion exchange resin column, concentrating under reduced pressure, and drying to obtain 0.20kg of momordica grosvenori flower total saponins;
(6) and (3) separating total flavonoids: eluting the anion exchange resin column after the total saponins are separated in the step (5) by using 2.5BV of sodium hydroxide aqueous solution with the mass percent concentration of 4 percent to obtain alkaline elution liquid; neutralizing the alkaline eluent with 5% sulfuric acid, desalting with nanofiltration membrane (the cut-off molecular weight of the nanofiltration membrane is 200Da, and the filtering pressure is 0.4 Mpa), collecting the nanofiltration membrane cut-off liquid, concentrating the nanofiltration membrane cut-off liquid under reduced pressure, and drying to obtain 0.53kg of total flavonoids of Siraitia grosvenorii Swingle.
According to the determination of the method specified in Chinese pharmacopoeia, the content of the volatile oil in the momordica grosvenori flower volatile oil product obtained in the embodiment is 91.62%, and the yield is 93.27%; through ultraviolet spectrophotometry, the content of the total saponins in the grosvenor momordica flower total saponins product obtained in the embodiment is 90.28%, and the yield is 94.04%; the content of the total flavonoids in the momordica grosvenori flower total flavonoids product obtained in the embodiment is 91.35% and the yield is 91.18% through ultraviolet spectrophotometry.
Example 3
The embodiment comprises the following steps:
(1) extracting volatile oil: crushing 10kg of dried momordica grosvenori flowers until the particle size is 2-5 mm, placing the crushed momordica grosvenori flowers in an extraction tank, adding 160L of mixed organic solvent (120 # solvent oil: ethyl acetate = 9: 1, V/V), extracting for 4 hours under reflux, and filtering to obtain organic solvent extract containing volatile oil and extraction residues containing total flavonoids and total saponins;
(2) and (3) volatile oil purification: adding 0.1kg of activated clay into the organic solvent extract obtained in the step (1), stirring and decoloring for 4 hours, filtering, collecting filtrate, and concentrating the filtrate to obtain 0.62kg of a momordica grosvenori flower volatile oil product;
(3) extracting total flavonoids and total saponins: distilling the organic solvent in the extraction residue obtained in the step (1) to be clean, then adding 200L of 70% methanol solution with volume concentration, refluxing and extracting for 4 hours, and filtering to obtain an alcohol extract;
(4) enrichment of total flavonoids and total saponins: concentrating the alcohol extract obtained in the step (3) under reduced pressure, adsorbing by using a macroporous adsorption resin column (the model of the macroporous adsorption resin is DM130, the using amount of the macroporous adsorption resin is 10L, the height-diameter ratio of the macroporous adsorption resin column is 4:1, and the flow rate of the macroporous adsorption resin column is 0.5 BV/h), eluting the macroporous adsorption resin column by using absolute ethyl alcohol, collecting, and concentrating the eluent to obtain a concentrate containing the total flavonoids and the total saponins of the momordica grosvenori flower;
(5) and (3) separation of total saponins: decoloring the concentrated solution of the total flavonoids and the total saponins obtained in the step (4) by using a macroporous anion exchange resin column (the model of the macroporous anion exchange resin is LSA-700, the dosage of the anion resin is 10L of the weight of the raw material momordica grosvenori flower, the height-diameter ratio of the anion exchange resin column is 4:1, the flow rate of the anion exchange resin column is 0.5 BV/h), collecting the effluent of the anion exchange resin column, concentrating under reduced pressure, and drying to obtain 0.19kg of the momordica grosvenori flower total saponins;
(6) and (3) separating total flavonoids: eluting 2BV of potassium hydroxide aqueous solution with the mass percent concentration of 4.5% by using the anion exchange resin column after the total saponins are separated in the step (5) to obtain alkaline elution liquid; neutralizing the alkaline eluent with 10% hydrochloric acid, desalting with nanofiltration membrane (the cut-off molecular weight of the nanofiltration membrane is 200Da, and the filtering pressure is 0.5 Mpa), collecting the nanofiltration membrane cut-off liquid, concentrating the nanofiltration membrane cut-off liquid under reduced pressure, and drying to obtain 0.58kg of flos Momordicae Charantiae total flavone product.
The content of the volatile oil in the momordica grosvenori flower volatile oil product obtained in the embodiment is 93.95% and the yield is 95.65% determined by a method of Chinese pharmacopoeia; through ultraviolet spectrophotometry, the content of the total saponins in the momordica grosvenori flower total saponin product obtained in the embodiment is 91.77%, and the yield is 90.81%; the content of the total flavonoids in the momordica grosvenori flower total flavonoids product obtained in the embodiment is 89.76% and the yield is 94.48% through ultraviolet spectrophotometry.
Claims (10)
1. The method for continuously extracting and separating a plurality of natural active ingredients from the momordica grosvenori flower is characterized by comprising the following steps of:
(1) extracting volatile oil: crushing dried momordica grosvenori flower, placing in an extraction tank, adding an organic solvent, performing reflux extraction, and filtering to obtain an organic solvent extract containing volatile oil and extraction residues containing total flavonoids and total saponins;
(2) and (3) volatile oil purification: adding a decoloring agent into the organic solvent extracting solution obtained in the step (1), stirring and decoloring, filtering, collecting filtrate, and concentrating the filtrate to obtain a momordica grosvenori flower volatile oil product;
(3) extracting total flavonoids and total saponins: distilling the organic solvent in the extraction residue obtained in the step (1) to be clean, then adding methanol or ethanol, carrying out reflux extraction, and filtering to obtain an alcohol extract;
(4) enrichment of total flavonoids and total saponins: concentrating the alcohol extract obtained in step (3) under reduced pressure, adsorbing with macroporous adsorbent resin column, eluting with ethanol, collecting eluate, and concentrating to obtain concentrated solution containing total flavone and total saponin;
(5) and (3) separation of total saponins: decoloring the concentrated solution containing the total flavonoids and the total saponins obtained in the step (4) by using a macroporous anion exchange resin column, collecting the effluent of the anion exchange resin column, concentrating under reduced pressure, and drying to obtain a total saponin product;
(6) and (3) separating total flavonoids: eluting the anion exchange resin column after the total saponins are separated in the step (5) by using alkali liquor to obtain alkali elution liquid; neutralizing the alkaline eluent with acid, desalting with nanofiltration membrane, collecting the nanofiltration membrane retentate, concentrating under reduced pressure, and drying to obtain total flavone product.
2. The method of claim 1, wherein the extraction and separation is performed continuously for a plurality of daysThe method for preparing the active ingredient is characterized in that in the step (1), the organic solvent is a mixed solvent of a low-polarity organic solvent and ethyl acetate; the low-polarity organic solvent is preferably diethyl ether, petroleum ether, hexane, cyclohexane, heptane, 6#Solvent oil and 120#One or more solvent oils; the volume ratio of the low-polarity organic solvent to the ethyl acetate is preferably 5-10: 1.
3. the method for continuously extracting and separating multiple natural active ingredients from momordica grosvenori flower according to claim 1 or 2, wherein in the step (1), the amount of the organic solvent is 10-20 times (L/kg) of the weight of the momordica grosvenori flower; the reflux extraction time is 2-4 hours.
4. The method for continuously extracting and separating multiple natural active ingredients from momordica grosvenori flower according to any one of claims 1 to 3, wherein in the step (2), the decolorizing agent is one or more of activated carbon, activated clay, diatomite, alumina adsorbent resin and silica gel adsorbent resin; the dosage of the decolorant is 0.1-2% of the weight of the raw material momordica grosvenori flower; and the stirring and decoloring time is 1-4 hours.
5. The method for continuously extracting and separating multiple natural active ingredients from momordica grosvenori flower according to any one of claims 1 to 4, wherein in the step (3), the volume fraction of the methanol or the ethanol is 50% to 90%; the dosage of the methanol or the ethanol is 10-20 times (L/kg) of the weight of the raw material momordica grosvenori flower; the reflux extraction time is 2-4 hours.
6. The method for continuously extracting and separating multiple natural active ingredients from momordica grosvenori flower according to any one of claims 1 to 5, wherein the type of the macroporous adsorption resin is D101, AB-8, LX-38 or DM 130; the dosage (volume) of the macroporous adsorption resin is 0.5-1 time (L/kg) of the weight of the raw material momordica grosvenori flower; the height-diameter ratio of the macroporous adsorption resin column is 4-6: 1; the flow rate of the upper column is 0.5-1.0 BV/h.
7. The method for continuously extracting and separating multiple natural active ingredients from momordica grosvenori flower according to one of claims 1 to 6, wherein in the step (5), the type of the macroporous anion exchange resin is D941, D945, LSA-700 or LSD 762; the dosage of the anion resin is 0.5-1 time (L/kg) of the weight of the raw material momordica grosvenori flower; the height-diameter ratio of the anion exchange resin column is preferably 4-6: 1; the flow rate of the upper column is preferably 0.5-1.0 BV/h.
8. The method for continuously extracting and separating multiple natural active ingredients from momordica grosvenori flower according to any one of claims 1 to 7, wherein in the step (6), the alkali solution is an aqueous solution of sodium hydroxide or potassium hydroxide; the mass percentage concentration of the alkali liquor is 2-5%; the dosage of the alkali liquor is 2-3 BV.
9. The method for continuously extracting and separating multiple natural active ingredients from momordica grosvenori flower according to any one of claims 1 to 8, wherein in the step (6), the acid is hydrochloric acid or sulfuric acid, and the concentration of the acid is preferably 5-15% by mass.
10. The method for continuously extracting and separating multiple natural active ingredients from momordica grosvenori flower according to any one of claims 1 to 9, wherein in the step (6), the molecular weight cut-off of the nanofiltration membrane is 100-500 Da; the filtration pressure is preferably 0.3-0.6 MPa.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115227624A (en) * | 2022-08-30 | 2022-10-25 | 北京臻味坊食品有限公司 | Infant skin care oil containing nut oil |
| WO2022247097A1 (en) * | 2021-05-28 | 2022-12-01 | 湖南绿蔓生物科技股份有限公司 | Method for separating active ingredients from siraitia grosvenorii flowers |
| CN116369529A (en) * | 2023-05-06 | 2023-07-04 | 邓婕 | Method for preparing vitamin E, C and stevioside V nutritional food from male flowers of fructus momordicae |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1375499A (en) * | 2002-03-28 | 2002-10-23 | 何伟平 | Method of extracting and separating several components from mangsteen |
| JP2014034552A (en) * | 2012-08-09 | 2014-02-24 | Kao Corp | Biofilm formation inhibitor |
| CN105462690A (en) * | 2015-11-18 | 2016-04-06 | 广州暨南生物医药研究开发基地有限公司 | Argy wormwood leaf essence oil and preparing method thereof |
| CN109053821A (en) * | 2018-09-30 | 2018-12-21 | 湖南华诚生物资源股份有限公司 | The method of tea polyphenols, total amino acid, chromocor compound is extracted from grosvenor momordica leaf |
| CN109674843A (en) * | 2019-01-30 | 2019-04-26 | 湖南绿蔓生物科技股份有限公司 | A kind of method for extraction and purification of dried fructus momordicae comprehensive utilization |
| CN111297936A (en) * | 2020-02-24 | 2020-06-19 | 湖南华诚生物资源股份有限公司 | A method for extracting and separating total flavonoids, total triterpenoid saponins and total polysaccharides from Luo Han Guo root |
| CN113041279A (en) * | 2021-05-28 | 2021-06-29 | 湖南绿蔓生物科技股份有限公司 | Method for separating effective components from momordica grosvenori flowers |
-
2021
- 2021-07-14 CN CN202110792921.6A patent/CN113398157A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1375499A (en) * | 2002-03-28 | 2002-10-23 | 何伟平 | Method of extracting and separating several components from mangsteen |
| JP2014034552A (en) * | 2012-08-09 | 2014-02-24 | Kao Corp | Biofilm formation inhibitor |
| CN105462690A (en) * | 2015-11-18 | 2016-04-06 | 广州暨南生物医药研究开发基地有限公司 | Argy wormwood leaf essence oil and preparing method thereof |
| CN109053821A (en) * | 2018-09-30 | 2018-12-21 | 湖南华诚生物资源股份有限公司 | The method of tea polyphenols, total amino acid, chromocor compound is extracted from grosvenor momordica leaf |
| CN109674843A (en) * | 2019-01-30 | 2019-04-26 | 湖南绿蔓生物科技股份有限公司 | A kind of method for extraction and purification of dried fructus momordicae comprehensive utilization |
| CN111297936A (en) * | 2020-02-24 | 2020-06-19 | 湖南华诚生物资源股份有限公司 | A method for extracting and separating total flavonoids, total triterpenoid saponins and total polysaccharides from Luo Han Guo root |
| CN113041279A (en) * | 2021-05-28 | 2021-06-29 | 湖南绿蔓生物科技股份有限公司 | Method for separating effective components from momordica grosvenori flowers |
Non-Patent Citations (7)
| Title |
|---|
| NARUNAN WUTTISIN，等: "Total Phenolic, Flavonoid Contents and Antioxidant Activity of Siraitia grosvenorii Fruits Extracts", 《FOOD AND APPLIED BIOSCIENCE JOURNAL》 * |
| 任秀如，等: "罗汉果花中皂苷的提取及抑菌活性研究", 《食品工业》 * |
| 刘庚贵，等: "基于大孔吸附树脂纯化罗汉果甜苷 V 的工艺优化研究", 《企业科技与发展》 * |
| 卞伟明，等: "罗汉果花黄酮和多糖的提取与活性研究", 《粮食与油脂》 * |
| 张卓睿,等: "响应面法对罗汉果花总黄酮超声波提取工艺的优化", 《湖北农业科学》 * |
| 梁志远，等: "不同提取方法对罗汉果花挥发油成分的影响", 《时珍国医国药》 * |
| 赵海燕，等: "利用D280阴离子交换树脂分离黄芪皂苷和黄酮的研究", 《安徽农业科学》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2022247097A1 (en) * | 2021-05-28 | 2022-12-01 | 湖南绿蔓生物科技股份有限公司 | Method for separating active ingredients from siraitia grosvenorii flowers |
| CN115227624A (en) * | 2022-08-30 | 2022-10-25 | 北京臻味坊食品有限公司 | Infant skin care oil containing nut oil |
| CN115227624B (en) * | 2022-08-30 | 2023-11-14 | 北京臻味坊食品有限公司 | Infant skin care oil containing nut oil |
| CN116369529A (en) * | 2023-05-06 | 2023-07-04 | 邓婕 | Method for preparing vitamin E, C and stevioside V nutritional food from male flowers of fructus momordicae |
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