CN111388514A - Method for extracting flavone from holboellia latifolia - Google Patents

Method for extracting flavone from holboellia latifolia Download PDF

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Publication number
CN111388514A
CN111388514A CN202010442324.6A CN202010442324A CN111388514A CN 111388514 A CN111388514 A CN 111388514A CN 202010442324 A CN202010442324 A CN 202010442324A CN 111388514 A CN111388514 A CN 111388514A
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flavone
holboellia latifolia
membrane
extracting
water
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汪守鹏
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105Plant extracts, their artificial duplicates or their derivatives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783Angiosperms [Magnoliophyta]
    • A61K8/9789Magnoliopsida [dicotyledons]
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10Preparation or pretreatment of starting material
    • A61K2236/19Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/33Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50Methods involving additional extraction steps
    • A61K2236/51Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50Methods involving additional extraction steps
    • A61K2236/53Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Botany (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Engineering & Computer Science (AREA)
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  • Biotechnology (AREA)
  • Animal Behavior & Ethology (AREA)
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  • General Health & Medical Sciences (AREA)
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  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Epidemiology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Medicinal Chemistry (AREA)
  • Medical Informatics (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Nutrition Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
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  • Preparation Of Compounds By Using Micro-Organisms (AREA)
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Abstract

The invention discloses a method for extracting flavone from holboellia latifolia, which comprises the following steps of adding cellulase, α -amylase and water into holboellia latifolia powder, stirring and extracting for 0.5h at 50-70 ℃, then adding pectinase, stirring for 10min at 50 ℃, standing for 24h, filtering to remove precipitates, taking supernate, clarifying and filtering the supernate, separating polysaccharide from flavone in the filtrate, and leaving flavone liquid and drying to obtain a light brown powdery product.

Description

Method for extracting flavone from holboellia latifolia
Technical Field
The invention relates to a method for extracting flavone, in particular to a method for extracting flavone from holboellia latifolia, and belongs to the technical field of extraction of effective components of plants.
Background
The holboellia latifolia is a wild fruit, mainly grows in the mountainous areas of Hunan and Hubei parts of China, and is also distributed in the China's southwest, east China, Chongqing, Yunnan, Shanxi, Gansu and other places.
The August melon and fruit is similar to banana, has succulent pulp and fragrant and sweet fruit taste, is rich in various vitamins, amino acids (including valine, methionine, isoleucine, phenylalanine, lysine and the like which can not be synthesized by human body), soluble fructose, starch, carbohydrate, organic acid protein and flavone, is also rich in elements such as calcium, phosphorus, iron, zinc, selenium and the like and various trace elements required by human body, is called as wild banana and is a pollution-free green food.
The holboellia latifolia has rich nutrition and high medicinal value, has the effects of clearing heat and promoting diuresis, promoting blood circulation and dredging collaterals, and promoting qi circulation and relieving pain, is mainly used for treating oliguria with reddish urine, stranguria with turbid urine, edema, rheumatic arthralgia, traumatic injury, galactostasis, hernia pain, uterine prolapse and orchitis, and is one of five major precious traditional Chinese medicinal materials which are mainly protected and developed by relevant departments of China.
According to reports, the flavonoid compound not only has the effects of improving microcirculation, expanding coronary arteries, increasing blood flow of brain and coronary arteries, slowing heart rate and reducing oxygen consumption of cardiac muscle, but also has the effects of resisting aging, inhibiting blood coagulation, resisting cancer, inhibiting bacteria and disinfecting, and has wide application in the industries of medicines, health products and foods.
The fruit of the holboellia latifolia is rich in flavone, and the existing method for extracting the flavone from the holboellia latifolia comprises the following steps:
alcohol extraction (n-butanol) -ethanol extraction of extract-crystallization of extract acetic acid-crystal drying.
In actual production, the existing extraction method has a plurality of problems, specifically:
1. the n-butyl alcohol and the acetic acid are high-boiling-point solvents, and need high-temperature drying, so that the energy consumption is high;
2. in the high-temperature drying process, gum peculiar smell can be generated, and the product quality is poor;
3. the content of flavone in the product is about 7 percent, and the content is lower and needs to be further improved;
4. the product has poor water solubility and chemical solvent residue, so the application is limited, and the market cannot be expanded.
Disclosure of Invention
In order to solve the defects of the prior art, the invention aims to provide a novel method for extracting flavone from holboellia latifolia, and the holboellia latifolia flavone extracted by the method has no peculiar smell, no chemical solvent residue, good water solubility and high content.
In order to achieve the above object, the present invention adopts the following technical solutions:
the method for extracting flavone from holboellia latifolia is characterized by comprising the following steps:
step 1, adding cellulase, α -amylase and water into the holboellia latifolia powder, stirring and extracting at 50-70 ℃ for 0.5h, then adding pectinase, stirring at 50 ℃ for 10min, and then standing for 24 h;
step 2: filtering to remove precipitate, collecting supernatant, clarifying and filtering the supernatant, and separating polysaccharide and flavone from the filtrate to obtain flavone solution;
and step 3: drying the flavone liquid to finally obtain a light brown powdery product.
The method for extracting flavone from holboellia latifolia is characterized in that 5g of cellulase, 12g of α -amylase, 20g of pectinase and 5L water are added into 1kg of holboellia latifolia powder in step 1.
The method for extracting flavone from holboellia latifolia is characterized in that in the step 2, the supernatant is clarified and filtered by using a membrane T-10 or a membrane T-7, and the filtrate is subjected to separation of polysaccharide and flavone by using a membrane G20 or a membrane G19.
The method for extracting flavone from holboellia latifolia is characterized in that in the step 3, the drying temperature is 60 ℃.
The invention has the advantages that:
1. the extraction process uses water as solvent, does not use organic solvent, and the product has no organic solvent residue;
2. the high-temperature drying process is not involved, and the product has no peculiar smell;
3. the water is used as a solvent in the extraction process, so that the water solubility of the product is good;
4. the crude fiber is enzymolyzed by cellulase, the starch and the protein are enzymolyzed by α -amylase, and the pectin is enzymolyzed by pectinase, wherein the three kinds of enzymolysis can improve the flavone storage rate (about 60 percent increase), so the total flavone content in the product is obviously improved.
Detailed Description
The present invention will be described in detail with reference to the following embodiments.
Example 1
Taking 1kg of dried holboellia latifolia thumb, crushing, soaking holboellia latifolia thumb powder in 5L water, decocting twice for 2h each time, separating liquid residues, combining filtrates, performing column chromatography on the filtrate by using nonpolar resin D101 (or neutral resin L SA-21), eluting with water at the chromatography speed of 1L/h, discarding the column eluate and the column eluate (containing monosaccharide and small molecular weight impurities) of water washing 2L, receiving water eluate (which is polysaccharide and flavone feed liquid) 10L, performing membrane concentration on the water eluate, drying the concentrated solution at the drying temperature of 60 ℃, and finally obtaining a light brown powdery product.
437g of the product is obtained by weighing, and the yield is 43.7%.
The detection shows that the content of the total flavone in the product is 7.1 wt%.
Example 2
Taking 1kg of dried holboellia latifolia, crushing, adding 5g of cellulase, 12g of α -amylase and 10L of water, stirring and extracting at 40 ℃ for 0.5h, then carrying out column chromatography by using nonpolar resin D101 (or neutral resin L SA-21), eluting with water at the chromatography speed of 1L/h, removing feed liquid on the upper water column and the lower liquid (containing monosaccharide and small molecular weight impurities) of the 2L column before water washing, then receiving water eluent (polysaccharide and flavone feed liquid) 10L, then carrying out membrane concentration on the water eluent, drying the concentrated solution at the drying temperature of 60 ℃, and finally obtaining a light brown powdery product.
After weighing, 679g of the product was obtained with a yield of 67.9%.
The detection shows that the content of the total flavone in the product is 11.3 wt%.
Example 3
Taking 1kg of dried holboellia latifolia, crushing, adding 5G of cellulase, 12G of α -amylase and 5L of water, stirring and extracting for 0.5h at 40 ℃, then adding 20G of pectinase, stirring for 10min at 40 ℃, standing for 24h, then filtering to remove precipitates, taking supernatant, clarifying and filtering by using a membrane T-10 (or a membrane T-7), separating polysaccharide and flavone by using a membrane G20 (or a membrane G19) for filtrate, remaining solution in the membrane, drying the solution, and drying at 60 ℃ to finally obtain a light brown powdery product.
467g of product is obtained by weighing, and the yield is 46.7%.
The detection shows that the content of the total flavone in the product is wtw 13.0.0%.
Example 4
Taking 1kg of dried holboellia latifolia, crushing, adding 5G of cellulase, 12G of α -amylase and 5L of water, stirring for 0.5h at 80 ℃, then adding 20G of pectinase, stirring for 10min at 80 ℃, standing for 24h, then filtering to remove precipitates, taking supernatant, clarifying and filtering by using a membrane T-10 (or a membrane T-7), separating polysaccharide and flavone by using a membrane G20 (or a membrane G19) for filtrate, remaining solution in the membrane, drying the solution, and drying at 60 ℃ to finally obtain a light brown powdery product.
367g of the product is obtained by weighing, and the yield is 36.7%.
The detection shows that the content of the total flavone in the product is 13.0%.
Example 5
Taking 1kg of dried holboellia latifolia, crushing, adding 5g of cellulase, 12g of α -amylase and 5g of water 5L, standing at 50 ℃ for 24h, filtering to remove precipitates, taking supernatant, adding 50% ethanol solution 2L into the supernatant, stirring for 10min, standing for 4h, recovering ethanol under reduced pressure, diluting and centrifuging the concentrated pasty fluid with water, leaving the supernatant, carrying out column chromatography on the supernatant by using non-polar resin D101 (or neutral resin L SA-21), at the chromatography speed of 1L/h, eluting with water, discarding residues, receiving water eluent and ethanol eluent, combining the water eluent and the ethanol eluent, recovering ethanol, drying the concentrated pasty fluid at the drying temperature of 60 ℃, and finally obtaining a light brown powdery product.
Weighing to obtain 678g of a product, wherein the yield is 67.8%.
Detection shows that the content of total flavone in the product is 57.0%, and ethanol remains.
Example 6
Taking 1kg of dried holboellia latifolia, crushing, adding 5G of cellulase, 12G of α -amylase and 5L of water, stirring for 0.5h at 50 ℃, then adding 20G of pectinase, stirring for 10min at 50 ℃, standing for 24h, then filtering to remove precipitates, taking supernatant, then clarifying and filtering by using a membrane T-10 (or a membrane T-7), separating polysaccharide and flavone by using a membrane G20 (or a membrane G19) for filtrate, leaving solution (flavone feed liquid) in the membrane, drying the solution, and drying at 60 ℃ to finally obtain a light brown powdery product.
After weighing, 937g of product is obtained, and the yield is 93.7%.
Detection shows that the content of total flavone in the product is 73.0%.
Example 7
Taking 1kg of dried holboellia latifolia, crushing, adding 5G of cellulase, 12G of α -amylase and 5L of water, stirring and extracting for 0.5h at 70 ℃, then adding 20G of pectinase, stirring for 10min at 50 ℃, standing for 24h, then filtering to remove precipitates, taking supernatant, then clarifying and filtering by using a membrane T-10 (or a membrane T-7), separating polysaccharide and flavone by using a membrane G20 (or a membrane G19) for filtrate, leaving solution (flavone feed liquid) in the membrane, drying the solution, and finally obtaining a light brown powdery product at the drying temperature of 60 ℃.
The product was weighed to yield 957g, 95.7% yield.
Detection shows that the content of total flavone in the product is 73.0%.
The yields of the products of example 5, example 6 and example 7 were all high and the total flavone content in the products was also high, so we performed the water solubility test again on these three samples.
The testing process comprises the following steps: the products of example 5, example 6 and example 7 were each formulated as aqueous solutions at a concentration of 10 w/v%, and are designated as sample 1, sample 2 and sample 3, respectively.
And (3) testing results: samples 2 and 3 were not suspended, precipitated and completely dissolved; sample 1 had a brown precipitate.
And (4) test conclusion: the water solubility of the products of examples 6 and 7 was better than that of the product of example 5.
The products of the embodiment 6 and the embodiment 7 have good water solubility and no ethanol residue, so the method can be applied to the production of cosmetics, foods, medicines and the like, the application range is greatly expanded, and the market prospect is good.
It should be noted that the above-mentioned embodiments do not limit the present invention in any way, and all technical solutions obtained by using equivalent alternatives or equivalent variations fall within the protection scope of the present invention.

Claims (5)

1. The method for extracting flavone from holboellia latifolia is characterized by comprising the following steps:
step 1, adding cellulase, α -amylase and water into the holboellia latifolia powder, stirring and extracting at 50-70 ℃ for 0.5h, then adding pectinase, stirring at 50 ℃ for 10min, and then standing for 24 h;
step 2: filtering to remove precipitate, collecting supernatant, clarifying and filtering the supernatant, and separating polysaccharide and flavone from the filtrate to obtain flavone solution;
and step 3: drying the flavone liquid to finally obtain a light brown powdery product.
2. The method for extracting flavone from holboellia latifolia as claimed in claim 1, wherein 5g of cellulase, 12g of α -amylase, 20g of pectinase and 5L water are added to 1kg of holboellia latifolia powder in step 1.
3. The method for extracting flavone from holboellia latifolia as claimed in claim 1, wherein in step 2, the supernatant is clarified and filtered by membrane T-10 or membrane T-7.
4. The method for extracting flavone from holboellia latifolia as claimed in claim 1, wherein in step 2, the filtrate is subjected to separation of polysaccharide and flavone with membrane G20 or membrane G19.
5. The method for extracting flavone from holboellia latifolia as claimed in claim 1, wherein the drying temperature is 60 ℃ in step 3.
CN202010442324.6A 2020-02-25 2020-05-22 Method for extracting flavone from holboellia latifolia Pending CN111388514A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111728933A (en) * 2020-08-15 2020-10-02 东莞市风雨科技有限公司 Concealer with spot-lightening and moisturizing functions and preparation process thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108658919A (en) * 2018-05-04 2018-10-16 铜仁学院 The method for purifying and separating of Akebia trifoliate koiaz Peels flavones
CN110357980A (en) * 2019-07-11 2019-10-22 铜仁学院 A kind of extraction separation method of Akebia trifoliate koiaz Peels polysaccharide

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108658919A (en) * 2018-05-04 2018-10-16 铜仁学院 The method for purifying and separating of Akebia trifoliate koiaz Peels flavones
CN110357980A (en) * 2019-07-11 2019-10-22 铜仁学院 A kind of extraction separation method of Akebia trifoliate koiaz Peels polysaccharide

Non-Patent Citations (1)

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Title
戴好富等: "《天然产物现代分离技术》", 31 December 2006, 中国农业大学出版社 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111728933A (en) * 2020-08-15 2020-10-02 东莞市风雨科技有限公司 Concealer with spot-lightening and moisturizing functions and preparation process thereof

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Application publication date: 20200710