CN116832073B - Preparation method of pseudo-ginseng flower extract - Google Patents

Preparation method of pseudo-ginseng flower extract Download PDF

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CN116832073B
CN116832073B CN202310791684.0A CN202310791684A CN116832073B CN 116832073 B CN116832073 B CN 116832073B CN 202310791684 A CN202310791684 A CN 202310791684A CN 116832073 B CN116832073 B CN 116832073B
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notoginseng
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ginseng
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CN116832073A (en
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张云生
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Jinqi Pharmaceuticals Co ltd
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
    • A61K36/258—Panax (ginseng)
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, 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
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/38—Other non-alcoholic beverages
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, 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
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation 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
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation 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
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331—Extraction 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, decoction
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K2236/30—Extraction of the material
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation 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
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation 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
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

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Abstract

The invention relates to the technical field of plant extraction, in particular to a preparation method of a pseudo-ginseng flower extract, which comprises the following steps: (1) decocting Notoginseng flower in water, filtering to obtain decoction; (2) Performing alcohol precipitation on the water decoction, discarding the precipitate, collecting filtrate, and drying to obtain dried powder of the pseudo-ginseng flowers; (3) Adding water into the dried powder of the pseudo-ginseng flower, inoculating strains, fermenting, and obtaining the pseudo-ginseng flower. The invention has the advantages of high total saponin content and Rg3 content and excellent oxidation resistance.

Description

Preparation method of pseudo-ginseng flower extract
Technical Field
The invention relates to the technical field of plant extraction, in particular to a preparation method of a pseudo-ginseng flower extract.
Background
The notoginseng flower is a dried flower bud of Panax notoginseng (Panax pseudoginseng) belonging to the genus Panax of the family Araliaceae, is crisp and fragile, has slight smell and slightly bitter taste, contains various saponins and flavonoids of Panax ginseng, and has the functions of clearing heat and liver, reducing blood lipid, lowering blood pressure, resisting cancer, improving myocardial oxygen supply capacity, enhancing organism immunity, and the like; the Notoginseng total saponins have inhibiting effect on central nervous system, and are effective in tranquilizing, relieving inflammation, improving microcirculation, and lowering blood pressure; can be used for treating hypertension, dizziness, tinnitus, and acute pharyngolaryngitis; the traditional administration method is mostly soaking water for drinking.
In addition, the Chinese pharmacopoeia (2010 edition) prescribes that the main root, the branch root and the rhizome of the underground part of the pseudo-ginseng are medicinal parts, while the pseudo-ginseng flower is a non-traditional medicinal part of the pseudo-ginseng, but researches show that the pseudo-ginseng flower has the effects of tranquilizing, diminishing inflammation, easing pain, reducing blood pressure and the like, and has the advantages of wide pharmacological effect, less side effect and lasting and stable treatment effect. Therefore, the pseudo-ginseng flower also has higher medicinal value.
Notoginseng is the high content part of notoginsenoside in whole plant, and tens of saponin components including ginsenoside Rb1, rb2, rb3, rc, rd, etc. have been found in it so far, wherein the content of ginsenoside Rb3 and Rc is the highest. In addition, the chemical components in the pseudo-ginseng flower also comprise flavonoids, polysaccharides, volatile oils, inorganic matters and the like.
The method for extracting saponin from Notoginseng flower is heating reflux method, soaking method, percolation method, microwave method, etc., but these methods have problems of long extraction time, low efficiency, and loss and change of saponin component caused by long-time heating.
Chinese patent application CN101199565A relates to effective active site of Notoginseng flower bud and its preparation method, and specifically adopts alcohol extraction-centrifugation-macroporous resin-concentration-drying and pulverizing method to extract effective active site-Notoginseng flower bud total saponin from flower bud of traditional Chinese medicine Notoginseng. The extraction method provided by the invention improves the purity of the total saponins of the pseudo-ginseng flower buds in the extract, and simultaneously gives consideration to the transfer rate and the yield of the extract, so that the extract of the effective active ingredients of the pseudo-ginseng flower buds with stable transfer rate and yield and higher purity is obtained. The total saponins of the pseudo-ginseng flower buds account for 50-100% of the total weight of the extract. However, the method is more severe in conditions, the ethanol with 50% -80% of extraction solvent and the AB-8 resin type of macroporous resin are limited, and the extraction effect of the method is still further improved.
Chinese patent application CN108157965A discloses a preparation method of a pseudo-ginseng flower extract, which comprises the steps of sterilizing by microwaves, inactivating enzymes, drying, carrying out biological enzymolysis to degrade pesticide residues, then carrying out ultrasonic extraction and thermal reflux extraction to obtain a pseudo-ginseng flower crude extract, further adsorbing the pesticide residues by a small-aperture resin column, carrying out nanofiltration treatment to remove the pesticide residues, and carrying out freeze drying to obtain the pseudo-ginseng flower extract. However, the purpose of this method is to extract various active ingredients, and the extraction effect of saponins, particularly rare saponins, is to be further improved.
Chinese patent application CN110898090a discloses a total sanchinoside, which contains sanchinoside Fc, ginsenoside Rc, ginsenoside Rb2, ginsenoside Rb3, ginsenoside Rd in any proportion, and its preparation method and application; the invention takes the notoginseng flower as the raw material, and prepares the notoginseng flower total saponin through the processes of ultra-high pressure extraction, filtration, concentration, drying and the like. However, the invention does not describe the effect of extracting rare ginsenoside Rg 3.
Therefore, it is necessary to develop a preparation method of notoginseng flower extract which can solve the above technical problems.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a preparation method of a pseudo-ginseng flower extract with high total saponin content, high Rg3 content and excellent oxidation resistance.
The invention is realized by the following technical scheme:
a preparation method of a pseudo-ginseng flower extract comprises the following steps:
(1) Decocting Notoginseng flower in water, and filtering to obtain decoction;
(2) Performing alcohol precipitation on the water decoction, discarding the precipitate, collecting filtrate, and drying to obtain dried powder of the pseudo-ginseng flowers;
(3) Adding water into the dried powder of the pseudo-ginseng flower, inoculating strains, fermenting, and obtaining the pseudo-ginseng flower.
Preferably, in the step (1), notoginseng flower is decocted in water for 2-3 times, filtered, concentrated and cooled to obtain decoction.
Preferably, in the step (1), notoginseng flowers are decocted with 8-10 times of water.
Preferably, ethanol is added into the boiled water in the step (2) for alcohol precipitation, the mixture is stirred uniformly, the mixture is filtered after standing, the precipitate is removed, the filtrate is collected, the filtrate is concentrated under reduced pressure, and the filtrate is dried and crushed to obtain the dried notoginseng flower powder.
More preferably, the boiled liquid in the step (2) is added with 3-5 times of ethanol.
Preferably, the strain in step (3) is at least one of bacillus subtilis, aspergillus oryzae and saccharomyces cerevisiae.
More preferably, the bacterial species is a mixture of bacillus subtilis, aspergillus oryzae and saccharomyces cerevisiae.
More preferably, the mass ratio of the bacillus subtilis, the aspergillus oryzae and the saccharomyces cerevisiae is 3-5:1-3:2-4.
Preferably, the medium composition used for the fermentation in step (3) is 5-10g/L glucose, 3-8g/L peptone and 1-3g/L yeast extract.
Preferably, the fermentation condition in the step (3) is pH 4-5, inoculum size 4-6% and fermentation time 70-80h.
More preferably, the preparation method comprises the steps of:
(1) Decocting flos Notoginseng with 8-10 times of water for 2-3 times, filtering, concentrating, and cooling to obtain decoction;
(2) Adding 3-5 times of ethanol into the water decoction for alcohol precipitation, stirring, standing, filtering, discarding precipitate, collecting filtrate, concentrating under reduced pressure, drying, and pulverizing to obtain Notoginseng flower dry powder;
(3) Adding water into the dried notoginseng flower powder, inoculating strains for fermentation, wherein the strains are a mixture of bacillus subtilis, aspergillus oryzae and saccharomyces cerevisiae, and the mass ratio of the three is 3-5:1-3:2-4; the culture medium adopted by fermentation comprises 5-10g/L glucose, 3-8g/L peptone and 1-3g/L yeast extract; the fermentation conditions are pH 4-5, inoculum size 4-6% and fermentation time 70-80h, and the product is obtained after fermentation.
The invention also relates to the pseudo-ginseng flower extract prepared by the method.
The invention also relates to a beverage, which comprises the pseudo-ginseng flower extract.
The invention also relates to a food, which comprises the pseudo-ginseng flower extract.
The beneficial effects of the invention are as follows:
according to the invention, a fermentation process is added on the basis of extraction, and the total saponin content and Rg3 content in the finally prepared pseudo-ginseng flower extract are obviously improved.
According to the invention, bacillus subtilis, aspergillus oryzae and saccharomyces cerevisiae are adopted for synergistic fermentation, so that the total saponin content and Rg3 content in the pseudo-ginseng flower extract are further improved, and the antioxidant activity of the fermented product is obviously improved.
Detailed Description
The invention will be further described with reference to specific embodiments, and advantages and features of the invention will become apparent from the description. These examples are merely exemplary and do not limit the scope of the invention in any way. It will be understood by those skilled in the art that various changes and substitutions of details and forms of the technical solution of the present invention may be made without departing from the spirit and scope of the present invention, but these changes and substitutions fall within the scope of the present invention.
The bacillus subtilis adopted in the embodiments of the invention is purchased from Shandong Wuyang biotechnology limited company, and the model WY001 is the product number 001; aspergillus oryzae is available from Shandong Xinzhuoyuan chemical Co., ltd., model 255; saccharomyces cerevisiae was purchased from Shandong Wilman pet food Co., ltd., model king65.
Example 1
A preparation method of a pseudo-ginseng flower extract comprises the following steps:
(1) Decocting flos Notoginseng with 8 times of water for 2 times, filtering, concentrating, and cooling to obtain water decoction;
(2) Adding 3 times of ethanol into the water decoction for alcohol precipitation, stirring uniformly, standing, filtering, discarding precipitate, collecting filtrate, concentrating the filtrate under reduced pressure, drying, and pulverizing to obtain Notoginseng flower dry powder;
(3) Adding water into the dried pseudo-ginseng flower powder, inoculating strains for fermentation, wherein the strains are a mixture of bacillus subtilis, aspergillus oryzae and saccharomyces cerevisiae, and the mass ratio of the three is 3:1:2; the culture medium adopted by fermentation comprises 5g/L glucose, 3g/L peptone and 1g/L yeast extract; the fermentation conditions are pH 4, inoculum size 4% and fermentation time 70h, and the fermentation is completed.
Example 2
A preparation method of a pseudo-ginseng flower extract comprises the following steps:
(1) Decocting flos Notoginseng with 10 times of water for 3 times, filtering, concentrating, and cooling to obtain water decoction;
(2) Adding 5 times of ethanol into the water decoction for alcohol precipitation, stirring uniformly, standing, filtering, discarding precipitate, collecting filtrate, concentrating the filtrate under reduced pressure, drying, and pulverizing to obtain Notoginseng flower dry powder;
(3) Adding water into the dried pseudo-ginseng flower powder, inoculating strains for fermentation, wherein the strains are a mixture of bacillus subtilis, aspergillus oryzae and saccharomyces cerevisiae, and the mass ratio of the three is 5:3:4; the culture medium adopted by fermentation comprises 10g/L glucose, 8g/L peptone and 3g/L yeast extract; the fermentation conditions are pH 5, inoculum size 6%, fermentation time 80h, and the preparation method is completed after fermentation.
Example 3
A preparation method of a pseudo-ginseng flower extract comprises the following steps:
(1) Decocting Notoginseng flower in 9 times of water for 3 times, filtering, concentrating, and cooling to obtain water decoction;
(2) Adding 4 times of ethanol into the water decoction for alcohol precipitation, stirring uniformly, standing, filtering, discarding precipitate, collecting filtrate, concentrating the filtrate under reduced pressure, drying, and pulverizing to obtain Notoginseng flower dry powder;
(3) Adding water into the dried pseudo-ginseng flower powder, inoculating strains for fermentation, wherein the strains are a mixture of bacillus subtilis, aspergillus oryzae and saccharomyces cerevisiae, and the mass ratio of the three is 4:2:3; the culture medium adopted by fermentation comprises 7g/L glucose, 6g/L peptone and 2g/L yeast extract; the fermentation condition is pH 4.5, the inoculation amount is 5%, the fermentation time is 75 hours, and the fermentation is carried out.
Comparative example 1
The difference from example 3 is that the strains in step (3) are bacillus subtilis and aspergillus oryzae, the mass ratio of the bacillus subtilis to the aspergillus oryzae is 2:1, and the rest conditions are the same.
Comparative example 2
The difference from example 3 is that the strains in step (3) are bacillus subtilis and saccharomyces cerevisiae, the mass ratio of the bacillus subtilis to the saccharomyces cerevisiae is 4:3, and the rest conditions are the same.
Comparative example 3
The difference from example 3 is that the strains in step (3) are Aspergillus oryzae and Saccharomyces cerevisiae, the mass ratio of the two is 2:3, and the rest conditions are the same.
Comparative example 4
The difference from example 3 is that the strain in step (3) is Bacillus subtilis, and the other conditions are the same.
Test example 1
Total saponins and Rg3 content test
And centrifuging fermentation liquor obtained after fermentation of each example and comparative example, taking supernatant, and evaporating to dryness to obtain a sample to be tested.
Determination of total saponins content specific references (Shen Lan, feng Yi, xu Desheng, et al, colorimetric determination of total saponins content in Notoginseng [ J ]. Chinese patent medicine, 2007,29 (9): 3.).
The Rg3 content test uses high performance liquid chromatography, and the HPLC method is used for determining the change of ginsenoside Rg3 content in the notoginseng fermentation product (Wang Qing, yuan Ping, mao Rengang, etc.), part 2 of the test method in Chinese journal of Chinese medicine, 2011,29 (9): 2.).
The test was repeated 3 times and the test results are shown in table 1.
TABLE 1
Sample of Total saponin content (%) Rg3 content (mg/g)
Example 1 13.7 d 4.2 c
Example 2 13.9 d 4.3 c
Example 3 14.0 d 4.3 c
Comparative example 1 9.7 b 3.2 b
Comparative example 2 11.5 c 3.6 b
Comparative example 3 7.1 a 2.6 a
Comparative example 4 10.2 b 3.3 b
The different letters in the same column represent each group p <0.05 with significant differences.
Test example 2
Antioxidant Activity test
And centrifuging fermentation liquor obtained after fermentation of each example and comparative example, taking supernatant, and evaporating to dryness to obtain a sample to be tested.
Diluting a sample to be tested with 50% ethanol to obtain a diluent, and adding 1.5mL of the diluent with different mass concentrations into 2.5mL of 6.5X10 respectively -5 In a mol/L DPPH ethanol solution (50% ethanol is adopted), carrying out light-shielding reaction for 20min, and measuring an absorbance value Ai at a wavelength of 517 nm; absorbance Ac was measured by adding 2.5mL DPPH ethanol solution to 1.5mL 50% ethanol as above; the absorbance Aj of 1.5mL of the dilution reacted with 2.5mL of 50% ethanol was measured. The experiment was repeated 3 times with Trolox as reference, and the radical clearance was calculated: clearance (%) = [1- (Ai-Aj)/Ac]X 100%. The DPPH radical scavenging ability of each sample was measured at a concentration (IC) at which the scavenging rate reached 50% 50 Value) representation.
The results are shown in Table 2.
TABLE 2
Sample of IC 50 (mg/mL)
Trolox 0.024 a
Example 1 0.251 b
Example 2 0.247 b
Example 3 0.243 b
Comparative example 1 0.397 c
Comparative example 2 0.346 c
Comparative example 3 0.675 d
Comparative example 4 0.368 c
The different letters represent each group p <0.05 with significant differences.
The foregoing detailed description is directed to one of the possible embodiments of the present invention, which is not intended to limit the scope of the invention, but is to be accorded the full scope of all such equivalents and modifications so as not to depart from the scope of the invention.

Claims (10)

1. A preparation method of a pseudo-ginseng flower extract is characterized by comprising the following steps:
(1) Decocting Notoginseng flower in water, and filtering to obtain decoction;
(2) Performing alcohol precipitation on the water decoction, discarding the precipitate, collecting filtrate, and drying to obtain dried powder of the pseudo-ginseng flowers;
(3) Adding water into dried powder of Notoginseng flower, inoculating strain, fermenting, and fermenting to obtain the final product;
the strain is a mixture of bacillus subtilis, aspergillus oryzae and saccharomyces cerevisiae;
the mass ratio of the bacillus subtilis to the aspergillus oryzae to the saccharomyces cerevisiae is 3-5:1-3:2-4.
2. The method according to claim 1, wherein in the step (1), notoginseng flowers are decocted with water for 2-3 times, filtered, concentrated and cooled to obtain a decoction.
3. The method according to claim 1, wherein in step (1), notoginseng flowers are decocted with 8-10 times of water.
4. The method of claim 1, wherein the decoction in step (2) is added with ethanol for alcohol precipitation, stirred uniformly, left to stand, filtered, the precipitate is discarded, the filtrate is collected, concentrated under reduced pressure, dried and crushed to obtain the dried notoginseng flower powder.
5. The method according to claim 4, wherein the boiled liquid in step (2) is added with 3-5 times of ethanol.
6. The process according to claim 1, wherein the medium composition used for the fermentation in step (3) is 5-10g/L glucose, 3-8g/L peptone and 1-3g/L yeast extract.
7. The method according to claim 1, wherein the fermentation conditions in the step (3) are pH 4-5, inoculum size 4-6% and fermentation time 70-80h.
8. Notoginseng flower extract prepared by the method according to any one of claims 1-7.
9. A beverage comprising the notoginseng flower extract of claim 8.
10. A food product comprising the pseudo-ginseng flower extract according to claim 8.
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102676627A (en) * 2012-06-05 2012-09-19 浙江大学 Microbial fermentation extraction method of total arasaponin
CN110898090A (en) * 2019-12-03 2020-03-24 昆明理工大学 Panax notoginseng flower total saponin and preparation method and application thereof
CN112608949A (en) * 2020-12-18 2021-04-06 西南林业大学 Preparation method and application of pseudo-ginseng flower extract
CN115299515A (en) * 2022-08-23 2022-11-08 昆明生物制造研究院有限公司 Pseudo-ginseng stem and leaf fermentation powder and preparation method and application thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102676627A (en) * 2012-06-05 2012-09-19 浙江大学 Microbial fermentation extraction method of total arasaponin
CN110898090A (en) * 2019-12-03 2020-03-24 昆明理工大学 Panax notoginseng flower total saponin and preparation method and application thereof
CN112608949A (en) * 2020-12-18 2021-04-06 西南林业大学 Preparation method and application of pseudo-ginseng flower extract
CN115299515A (en) * 2022-08-23 2022-11-08 昆明生物制造研究院有限公司 Pseudo-ginseng stem and leaf fermentation powder and preparation method and application thereof

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