CN117357567A - Abelmoschus manihot seed extract, preparation method thereof and application thereof in treatment of respiratory diseases - Google Patents

Abelmoschus manihot seed extract, preparation method thereof and application thereof in treatment of respiratory diseases Download PDF

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CN117357567A
CN117357567A CN202311566970.3A CN202311566970A CN117357567A CN 117357567 A CN117357567 A CN 117357567A CN 202311566970 A CN202311566970 A CN 202311566970A CN 117357567 A CN117357567 A CN 117357567A
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extract
malva
seed
malva seed
extraction
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CN117357567B (en
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谢洋
张彭
徐百川
李得草
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First Affiliated Hospital of Henan University of TCM
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    • A—HUMAN NECESSITIES
    • 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)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
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    • 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
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • A—HUMAN NECESSITIES
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    • 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 provides a fraction-enriched extract of malva seeds, which comprises a water extract of malva seeds, an oil extract of malva seeds and an ethanol extract of malva seeds in a weight ratio of 5-8: 1 to 3: 3-6, wherein the extract of the malva seed which is enriched in a grading way has a synergistic effect, and the action effect is remarkably improved.

Description

Abelmoschus manihot seed extract, preparation method thereof and application thereof in treatment of respiratory diseases
Technical Field
The invention belongs to the field of natural medicines, and relates to an extract of semen Abutili, a preparation method thereof and application thereof in treating respiratory diseases.
Background
Abelmoschus manihot seed is fruit of Abelmoschus manihot Malva verticillata L and Abelmoschus manihot Mealva crispa L. The wild sunflower is distributed in all places in China. The malva is planted in the places of southwest and northwest of China, gansu, jiangxi, hubei and Hunan. Has effects of promoting diuresis, relieving stranguria, smoothing intestine, relaxing bowels, and promoting lactation. It is commonly used for gonorrhea, edema, constipation and milk failure.
Li Zengchun in "analysis of chemical composition of volatile oil of Mongolian medicine and Abelmoschus esculentus fruit" (Chinese patent medicine, page 922-924 in period 6 of 2008), 24 components are separated and identified by gas chromatography-mass spectrometry-computer retrieval technique, and the total peak area of volatile oil chromatography is 70.10%, wherein 6 compounds with content more than 4% are contained.
Patent AU2022201398A1 discloses an antioxidant and anti-inflammatory composition comprising an extract of okra as an active ingredient. The antioxidant effect of inhibiting the production of nitric oxide 5 by concentration dependency and the anti-inflammatory effect of inhibiting the expression of mRNA and protein of iNOS and COX-2 by concentration dependency were confirmed, and the extract of Abelmoschus manihot was effectively used as an antioxidant and anti-inflammatory pharmaceutical composition, a health functional food composition or a cosmetic composition.
The prior art has no report of treating respiratory diseases by using single component extract of the malva seed.
Therefore, intensive research on extraction of the active ingredients of the malva seeds is carried out, the functional ingredients in the malva seeds are enriched through a multistage extraction method, the value of the extract is improved, and multistage synergistic effect of the functional ingredients is effectively exerted, so that the necessity is very high.
Disclosure of Invention
The invention provides a fraction enriched extract of malva seeds, which is prepared by extracting and enriching functional components in the malva seeds in a fractional manner, and expands the application range and effect of the fraction enriched extract in preparing medicines for treating respiratory diseases by utilizing the synergistic effect of different functional components in treating respiratory diseases.
In order to achieve the above purpose, the invention adopts the following technical scheme:
the invention provides a fraction-enriched extract of malva seeds, which comprises a water extract of malva seeds, an oil extract of malva seeds and an ethanol extract of malva seeds in a weight ratio of 5-8: 1 to 3:3 to 6.
Further, the extract obtained by fractional enrichment of the malva seed is prepared from the ethanol extract of the malva seed, the oil extract of the malva seed and the ethanol extract of the malva seed according to the weight ratio of 6-8: 1 to 3: 3-5.
Further, the extract obtained by fractional enrichment of the malva seed is prepared from the ethanol extract of the malva seed, the oil extract of the malva seed and the ethanol extract of the malva seed according to the weight ratio of 7:2: 4.
The preparation method of the water extract of the malva seed comprises the following steps: the method comprises the steps of mixing the powder of the malva seed with 1g of the feed liquid: mixing 5-15 mL of water, regulating the pH value to 5-7, controlling the temperature to 80-90 ℃, performing ultrasonic extraction for 5-10min and stirring extraction for 10-20min to obtain periodic cyclic extraction, performing total extraction for 120-180min, performing solid-liquid separation after extraction is finished, and drying the leaching liquor to obtain a water extract of the malva seed, wherein filter residues are the powder 1 of the water extract of the malva seed;
in the preparation method of the water extract of the malva seed, a pH regulator is used for regulating the pH, wherein the pH regulator is citric acid, and the pH is preferably 5.5-6.5.
In the preparation method of the water extract of the malva seed, the ratio of the feed liquid to the water extract of the malva seed is preferably 1g to 8-10mL; the extraction time is preferably 140-160min.
In the preparation method of the water extract of the malva seed, the periodic cycle extraction is preferably carried out according to ultrasonic extraction for 6min and stirring extraction for 15 min; the extraction time is preferably 140-160min.
The preparation method of the malva seed oil extract comprises the following steps: the method comprises the steps of mixing the extract powder 1 of the malva seed according to a feed liquid ratio of 1g: mixing 15-20 mL of petroleum ether, extracting at 40-70 ℃ for 60-120 min under stirring, performing solid-liquid separation after extraction, and drying the leaching solution to obtain a malva seed oil extract, wherein filter residues are malva seed extract powder 2;
in the preparation method of the malva seed oil extract, the petroleum ether is preferably petroleum ether 60-90.
In the preparation method of the malva seed oil extract, the feed liquid ratio is preferably 1g:16-18mL, and the extraction time is preferably 90-100min.
The preparation method of the ethanol extract of the malva seed comprises the following steps: the method comprises the steps of mixing the extract powder 2 of the malva seed according to a feed liquid ratio of 1g: mixing 20-25 mL of ethanol water solution, extracting at 70-90 ℃ for 2-7min according to ultrasonic extraction, stirring and extracting for 10-15min for a period of cyclic extraction, extracting for 90-150min, carrying out solid-liquid separation after extraction, and drying the leaching liquor to obtain the ethanol extract of the malva seed.
In the preparation method of the ethanol extract of the malva seed, the ethanol aqueous solution is 20-40% ethanol aqueous solution.
In the preparation method of the ethanol extract of the malva seed, the ratio of the feed liquid to the ethanol extract of the malva seed is preferably 1g:22-24 mL.
In the preparation method of the ethanol extract of the malva seed, the periodic cycle extraction is preferably carried out according to ultrasonic extraction for 5min and stirring extraction for 12 min; the extraction time is preferably 110-130min.
The raw material adopted by the invention is the powder of the malva seed, which is obtained by cleaning, hot air drying and crushing the malva seed.
The research of the invention shows that the extract of the malva seed which is enriched in a grading way, namely, the water extract of the malva seed, the oil extract of the malva seed and the ethanol extract of the malva seed, has the weight ratio of 5-8: 1 to 3: 3-6, has excellent anti-respiratory disease effect, and compared with any single component, the composition provided by the invention generates synergistic effect, and the anti-respiratory disease effect is obviously improved.
The invention also aims to provide the application of the extract obtained by fractional enrichment of the malva seed in preparing medicines for treating respiratory diseases.
Furthermore, the invention provides application of the extract of the malva seed in preparing antiasthmatic drugs.
Compared with the prior art, the invention has the following advantages:
(1) The invention adopts a specific process to efficiently extract natural functional components of the malva seeds for treating respiratory diseases in a grading way, the active components of the malva seeds are utilized to the maximum extent, and the prepared multi-stage extract of the malva seeds has excellent respiratory disease treatment effect.
(2) The invention prepares the extract of the malva seed which is enriched in a grading way according to a specific weight ratio, and the three extracts have a synergistic effect, so that the effect is obviously improved.
Detailed Description
The invention discloses an extract of semen Abutili, a preparation method thereof and application thereof in treating respiratory diseases, and a person skilled in the art can refer to the content of the invention and combine the related principles of medicine extraction to properly improve the technological parameters. It is expressly noted that all such similar substitutions and modifications will be apparent to those skilled in the art, and are deemed to be included within the scope of the present invention.
For a better understanding of the present invention, and not to limit its scope, all numbers expressing quantities, percentages, and other values used in the present application are to be understood as being modified in all instances by the term "about". Each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.
The experimental methods used in the following examples are all conventional methods unless otherwise specified; the reagents, materials, etc. used in the examples described below are commercially available unless otherwise specified.
Example 1: water extract of semen Abutili
1Kg of malva seed powder is mixed according to a feed liquid ratio of 1g:8mL of water is mixed, the pH value is regulated to 5.7 by using citric acid solution, the temperature is controlled to be 80-90 ℃, the ultrasonic extraction is carried out for 6min and the stirring extraction is carried out for 15min for a period of cyclic extraction, the ultrasonic power is 300W, the total extraction is 147min, the solid-liquid separation is carried out after the extraction is finished, the leaching solution is dried, and the water extract of the malva seed is prepared, and the filter residue is the powder of the extract of the malva seed.
Example 2: water extract of semen Abutili
1Kg of malva seed powder is mixed according to a feed liquid ratio of 1g: mixing 10mL of water, regulating the pH value to 6.0 by using a citric acid solution, controlling the temperature to 80-90 ℃, circularly extracting according to ultrasonic extraction for 6min and stirring extraction for 15min for a period of time, wherein the ultrasonic power is 300W, extracting for 147min, carrying out solid-liquid separation after extraction, and drying the leaching liquor to obtain the water extract of the malva seed, wherein filter residues are the powder of the water extract of the malva seed.
Example 3: water extract of semen Abutili
1Kg of malva seed powder is mixed according to a feed liquid ratio of 1g:9mL of water is mixed, the pH value is regulated to 6.0 by using citric acid solution, the temperature is controlled to be 80-90 ℃, the ultrasonic extraction is carried out for 6min, the stirring extraction is carried out for 16min, the periodic cycle extraction is carried out, the ultrasonic power is 280W, the total extraction is carried out for 132min, the solid-liquid separation is carried out after the extraction is finished, the leaching liquor is dried, and the water extract of the malva seed is prepared, and the filter residue is the powder of the extract of the malva seed.
Example 4: water extract of semen Abutili
1Kg of malva seed powder is mixed according to a feed liquid ratio of 1g: mixing 10mL of water, regulating the pH value to 6.0 by using a hydrochloric acid solution, controlling the temperature to 80-90 ℃, circularly extracting according to ultrasonic extraction for 6min and stirring extraction for 15min for a period of time, wherein the ultrasonic power is 300W, extracting for 147min, carrying out solid-liquid separation after extraction, and drying the leaching liquor to obtain the water extract of the malva seed, wherein filter residues are the powder of the water extract of the malva seed.
Example 5: water extract of semen Abutili
1Kg of malva seed powder is mixed according to a feed liquid ratio of 1g: mixing 10mL of water, regulating the pH value to 6.0 by using a citric acid solution, controlling the temperature to 80-90 ℃, stirring and extracting for 147min, carrying out solid-liquid separation after the extraction is finished, and drying the leaching liquor to obtain a water extract of the malva seed, wherein filter residues are the powder of the extract of the malva seed;
example 6: water extract of semen Abutili
1Kg of malva seed powder is mixed according to a feed liquid ratio of 1g: mixing 10mL of water, regulating the pH value to 6.0 by using a citric acid solution, controlling the temperature to 80-90 ℃, performing ultrasonic extraction for 147min, performing ultrasonic power to 300W, performing solid-liquid separation after extraction is finished, and drying the leaching solution to obtain a water extract of the malva seed, wherein filter residues are the powder of the water extract of the malva seed;
example 7: water extract of semen Abutili
1Kg of malva seed powder is mixed according to a feed liquid ratio of 1g: mixing 10mL of water, regulating the pH value to 6.0 by using a citric acid solution, controlling the temperature to be 80-90 ℃, performing ultrasonic extraction for 3min and stirring extraction for 18min to perform periodic cyclic extraction, extracting for 147min, performing ultrasonic power to be 300W, performing solid-liquid separation after extraction is finished, and drying the leaching liquor to obtain a water extract of the malva seed, wherein filter residues are powder of the water extract of the malva seed;
example 8: water extract of semen Abutili
1Kg of malva seed powder is mixed according to a feed liquid ratio of 1g: mixing 10mL of water, regulating the pH value to 6.0 by using a citric acid solution, controlling the temperature to 80-90 ℃, performing ultrasonic extraction for 15min and stirring extraction for 6min to obtain periodic cyclic extraction, wherein the ultrasonic power is 300W, extracting for 147min, performing solid-liquid separation after extraction is finished, and drying the leaching liquor to obtain a water extract of the malva seed, wherein filter residues are powder of the water extract of the malva seed;
example 9: water extract of semen Abutili
The amount of the powder of the malva seed to be fed is 40Kg, and the water extract of the malva seed is prepared according to the method of example 1 to obtain the powder 1 of the extract of the malva seed of filter residue.
Example 10: abelmoschus manihot seed oil extract
1Kg of the extract powder 1 of the malva seed (prepared in example 9) was mixed with 1g of the extract powder: mixing 16mL petroleum ether 60-90, extracting at 50-60deg.C for 90min under stirring, separating solid and liquid after extraction, and drying the extractive solution to obtain semen Abutili oil extract, wherein the residue is semen Abutili extract powder;
example 11: abelmoschus manihot seed oil extract
1Kg of the extract powder 1 of the malva seed (prepared in example 9) was mixed with 1g of the extract powder: mixing 18mL of petroleum ether 60-90, extracting at 50-60deg.C for 100min under stirring, separating solid and liquid after extraction, and drying the leaching solution to obtain semen Abutili oil extract, wherein the filter residue is semen Abutili extract powder;
example 12: abelmoschus manihot seed oil extract
1Kg of the extract powder 1 of the malva seed (prepared in example 9) was mixed with 1g of the extract powder: mixing 16mL of normal hexane, extracting at 50-60 ℃ for 90min under stirring, performing solid-liquid separation after extraction, and drying the leaching solution to obtain a malva seed oil extract, wherein filter residues are malva seed extract powder;
example 13: abelmoschus manihot seed oil extract
1Kg of the extract powder 1 of the malva seed (prepared in example 9) was mixed with 1g of the extract powder: mixing 16mL petroleum ether 60-90, extracting at 50-60deg.C for 50min under stirring, separating solid and liquid after extraction, and drying the extractive solution to obtain semen Abutili oil extract, wherein the residue is semen Abutili extract powder;
example 14: abelmoschus manihot seed oil extract
The extract powder 1 of the malva seed was charged with 20Kg, and the oil extract of the malva seed was prepared as in example 10 to obtain a residue as the extract 2 of the malva seed.
Example 15: abelmoschus manihot seed ethanol extract
1Kg of the extract powder 2 of malva seed (prepared in example 14) was mixed with 1g of: 22mL of 40% ethanol aqueous solution is mixed, the extraction temperature is 70-90 ℃, the ultrasonic extraction is carried out for 5min, the stirring extraction is carried out for 12min, the periodic cycle extraction is carried out, the ultrasonic power is 200W, the total extraction is 119min, the solid-liquid separation is carried out after the extraction is finished, and the leaching solution is dried to obtain the ethanol extract of the malva seed.
Example 16: abelmoschus manihot seed ethanol extract
The preparation method comprises the steps of mixing the extract powder 2 of the malva seed (prepared in example 14) according to a feed liquid ratio of 1g:24mL of 20% ethanol aqueous solution is mixed, the extraction temperature is 70-90 ℃, the ultrasonic extraction is carried out for 5min, the stirring extraction is carried out for 12min, the periodic cycle extraction is carried out, the ultrasonic power is 200W, the total extraction is 119min, the solid-liquid separation is carried out after the extraction is finished, and the leaching solution is dried to obtain the ethanol extract of the malva seed.
Example 17: abelmoschus manihot seed ethanol extract
The preparation method comprises the steps of mixing the extract powder 2 of the malva seed (prepared in example 14) according to a feed liquid ratio of 1g:22mL of 30% ethanol aqueous solution is mixed, the extraction temperature is 70-90 ℃, the ultrasonic extraction is carried out for 5min, the stirring extraction is carried out for 12min, the periodic cycle extraction is carried out, the ultrasonic power is 200W, the total extraction is 119min, the solid-liquid separation is carried out after the extraction is finished, and the leaching solution is dried to obtain the ethanol extract of the malva seed.
Example 18: abelmoschus manihot seed ethanol extract
The preparation method comprises the steps of mixing the extract powder 2 of the malva seed (prepared in example 14) according to a feed liquid ratio of 1g:22mL of 80% ethanol aqueous solution is mixed, the extraction temperature is 70-80 ℃, the ultrasonic extraction is carried out for 5min, the stirring extraction is carried out for 12min, the periodic cycle extraction is carried out, the ultrasonic power is 200W, the total extraction is 119min, the solid-liquid separation is carried out after the extraction is finished, and the leaching solution is dried to obtain the ethanol extract of the malva seed.
Example 19: abelmoschus manihot seed ethanol extract
The preparation method comprises the steps of mixing the extract powder 2 of the malva seed (prepared in example 14) according to a feed liquid ratio of 1g:22mL of 10% ethanol aqueous solution is mixed, the extraction temperature is 70-80 ℃, the ultrasonic extraction is carried out for 5min, the stirring extraction is carried out for 12min, the periodic cycle extraction is carried out, the ultrasonic power is 200W, the total extraction is 119min, the solid-liquid separation is carried out after the extraction is finished, and the leaching solution is dried to obtain the ethanol extract of the malva seed.
Example 20: abelmoschus manihot seed ethanol extract
The preparation method comprises the steps of mixing the extract powder 2 of the malva seed (prepared in example 14) according to a feed liquid ratio of 1g:22mL of 30% ethanol aqueous solution is mixed, the extraction temperature is 70-90 ℃, stirring extraction is carried out for 119min, solid-liquid separation is carried out after extraction is finished, and the leaching solution is dried to obtain the ethanol extract of the malva seed.
Example 21: abelmoschus manihot seed ethanol extract
The preparation method comprises the steps of mixing the extract powder 2 of the malva seed (prepared in example 14) according to a feed liquid ratio of 1g:22mL of 30% ethanol aqueous solution is mixed, the extraction temperature is 70-90 ℃, the ultrasonic extraction is carried out for 12min, the stirring extraction is carried out for 5min, the periodic cycle extraction is carried out, the ultrasonic power is 100W, the total extraction is 119min, the solid-liquid separation is carried out after the extraction is finished, and the leaching solution is dried to obtain the ethanol extract of the malva seed.
Example 22: fractional enrichment extract of malva seed
The water extract of the malva seed, the oil extract of the malva seed and the ethanol extract of the malva seed are mixed according to the following proportion to prepare the extract of the malva seed with graded enrichment.
Example 23: test of antiasthmatics
1 materials and methods
1.1 animals
Male healthy guinea pigs (220-260 g in mass) are adaptively bred for 1 week, the room temperature is about 22-25 ℃, and the relative humidity is 40-70%.
1.2 test drugs
The fraction enriched extract of the malva seed prepared in example 22, namely the fraction enriched extract of the malva seed prepared in examples 22-1 to 22-14.
1.3 grouping and modeling
The model groups were randomly divided into 10 model groups, examples 22-1 through 22-14. All mice were subjected to molding.
And (3) molding: model group, example 22-1 to example 22-14, 30 mg/kg was intraperitoneally injected on the first day -1 The next day, 2mg of egg proteins and 100mg of aluminum hydroxide are prepared into 1ml of suspension for intraperitoneal injection, and after 21 days, 1ml of suspension prepared from egg proteins and 100mg of aluminum hydroxide is again injected intraperitoneally, and the suspension is injected again to induce sensitization of guinea pigs, and then the guinea pigs are placed in a 4L glass bell jar, and then the egg proteins with concentration of 1% are inhaled for 30s for inducing asthma.
1.4 modes of administration
Starting on day 1 after asthma induction by aerosol inhalation of 1% egg protein solution to guinea pigs, the groups of examples 22-1 to 22-11 were given equal doses of physiological saline for gastric lavage for 10 consecutive days, 1 time a day, according to the dose of 25mg/kg of extract.
After 1h of each intragastric administration, guinea pigs were stimulated with 1% egg proteins and asthma latency was recorded.
Asthma latency is the time required from inhalation of egg proteins after each episode to the onset of asthma. Asthma is judged by symptoms such as rapid respiratory rate, nodding respiratory asthma, nasal flaring, convulsion and the like.
2. Experimental results
Group of Asthma latency/second
Model group 50.67±12.77
Example 22-1 129.27±16.67##
Example 22-2 127.07±18.17##
Example 22-3 117.97±19.37##
Example 22-4 104.77±13.57##△
Examples 22 to 5 99.17±15.27##△
Examples 22 to 6 102.67±13.27##△
Examples 22 to 7 99.77±13.87##△
Examples 22 to 8 96.07±10.67##△
Examples 22 to 9 75.77±10.77#△△
Examples 22 to 10 80.47±12.87#△△
Examples 22 to 11 72.97±9.27#△△
Examples 22 to 12 58.17±10.87△△
Examples 22 to 13 54.07±9.67△△
Examples 22 to 14 55.37±8.97△△
Note that: comparing to the model group, #p <0.05, #p <0.01;
compared to the group of example 22-1, ΔP <0.05, ΔP <0.01.
(1) Compared with a model control group, the malva seed hierarchical enrichment extracts prepared in the examples 22-1 to 22-3 can remarkably prolong the incubation period of asthma, and all have very remarkable differences (P is smaller than 0.01), which indicates that the malva seed hierarchical enrichment extracts prepared in the examples 22-1 to 22-3 can treat asthma.
(2) Compared with a model control group, the malva seed hierarchical enrichment extracts prepared in the examples 22-4 to 22-5 can remarkably prolong the incubation period of asthma, and have very remarkable differences (P is smaller than 0.01), which indicates that the malva seed hierarchical enrichment extracts prepared in the examples 22-4 to 22-5 can treat asthma.
Compared with the embodiment 22-1, the malva seed fractional enrichment extracts prepared in the embodiments 22-4 to 22-5 are different in weight ratio of the water extract of the malva seed, the oil extract of the malva seed and the ethanol extract of the malva seed; the asthma latency period was significantly shortened in the groups of examples 22-4 to 22-5 compared to the group of example 22-1, each with a significant difference (P < 0.05), i.e., the fraction enriched extract of the malva seed prepared in the examples 22-4 to 22-5 was significantly different from the fraction enriched extract of the malva seed prepared in the example 22-1 in terms of treating asthma.
(3) Compared with a model control group, the malva seed hierarchical enrichment extracts prepared in the examples 22-6 to 22-7 can remarkably prolong the incubation period of asthma, and have very remarkable differences (P is smaller than 0.01), which indicates that the malva seed hierarchical enrichment extracts prepared in the examples 22-6 to 22-7 can treat asthma.
Compared with the embodiment 22-1, the malva seed fractionated enriched extracts prepared in the embodiments 22-6 to 22-7 are different in that the extraction process of the water extract of the malva seed is different; the asthma latency period was significantly shortened in the groups of examples 22-6 to 22-7 compared to the group of example 22-1, each having a significant difference (P < 0.05), i.e., the fraction enriched extract of the malva seed prepared in the groups of examples 22-6 to 22-7 was significantly different from the fraction enriched extract of the malva seed prepared in the group of example 22-1 in treating asthma.
(4) Compared with a model control group, the malva seed hierarchical enrichment extract prepared in the embodiment 22-8 can remarkably prolong the incubation period of asthma, and has very remarkable difference (P is less than 0.01), which indicates that the malva seed hierarchical enrichment extract prepared in the embodiment 22-8 can treat asthma.
The fraction enriched extract of the malva seed prepared in example 22-8 was different in the ethanol extract extraction process of the malva seed from example 22-1, and the asthma latency period of example 22-8 was significantly shortened compared to example 22-1, with a significant difference (P < 0.05), i.e., the fraction enriched extract of the malva seed prepared in example 22-8 was significantly different from the fraction enriched extract of the malva seed prepared in example 22-1 in terms of treating asthma.
(5) Compared with a model control group, the malva seed hierarchical enrichment extracts prepared in the examples 22-9 to 22-11 can remarkably prolong the incubation period of asthma, and have remarkable differences (P is less than 0.05), which indicates that the malva seed hierarchical enrichment extracts prepared in the examples 22-9 to 22-11 can treat asthma.
Compared with the embodiment 22-1, the malva seed fractional enrichment extracts prepared in the embodiments 22-9 to 22-11 are different in that the water extraction, the oil extraction and the ethanol extraction processes of the malva seed are different; the asthma latency period was significantly shortened compared to the group of example 22-1, and all of examples 22-9 to 22-11 were significantly different (P < 0.01), i.e., the fraction enriched extract of semen Abutili prepared in examples 22-9 to 22-11 was significantly different from the fraction enriched extract of semen Abutili prepared in example 22-1 in terms of treating asthma.
(6) The water extract of the malva seed of examples 22-12 was able to prolong the asthma latency, but did not have significant differences (P > 0.05) compared to the model control group, indicating that the water extract of the malva seed of examples 22-12 was not able to treat asthma.
(7) The Abelmoschus manihot oil extract of examples 22-13 was able to prolong asthma latency, but did not have significant differences (P > 0.05) compared to the model control group, indicating that the Abelmoschus manihot oil extract of examples 22-13 was not able to treat asthma.
(8) The ethanol extract of the malva seed of examples 22-14 was able to prolong the latency of asthma but did not have significant differences (P > 0.05) compared to the model control group, indicating that the oil extract of the malva seed of examples 22-14 was not able to treat asthma.
In conclusion, only the combination of the water extract of the malva seed, the oil extract of the malva seed and the ethanol extract of the malva seed can produce a synergistic effect to treat respiratory diseases; furthermore, only the combination of the water extract of the malva seed, the oil extract of the malva seed and the ethanol extract of the malva seed in a specific weight ratio can better produce a synergistic effect and better treat respiratory diseases.

Claims (10)

1. The extract is prepared from water extract of the malva seed, oil extract of the malva seed and ethanol extract of the malva seed according to the weight ratio of 5-8: 1 to 3:3 to 6.
2. The extract of the fractional enrichment of the malva seed according to claim 1, wherein the extract of the fractional enrichment of the malva seed is prepared from the ethanol extract of the malva seed, the oil extract of the malva seed and the ethanol extract of the malva seed according to the weight ratio of 6-8: 1 to 3: 3-5.
3. The extract of the fractional enrichment of the malva seed according to claim 1, wherein the extract of the fractional enrichment of the malva seed comprises the ethanol extract of the malva seed, the oil extract of the malva seed and the ethanol extract of the malva seed in a weight ratio of 7:2: 4.
4. The extract of the fractional enrichment of the malva seed according to claim 1, wherein the preparation method of the water extract of the malva seed comprises: the method comprises the steps of mixing the powder of the malva seed with 1g of the feed liquid: mixing 5-15 mL of water, regulating the pH value to 5-7, controlling the temperature to 80-90 ℃, performing ultrasonic extraction for 5-10min and stirring extraction for 10-20min to obtain periodic cyclic extraction, performing total extraction for 120-180min, performing solid-liquid separation after extraction is finished, and drying the leaching liquor to obtain the water extract of the malva seed, wherein filter residues are the powder 1 of the water extract of the malva seed.
5. The extract of the fractional enrichment of the malva seed according to claim 4, wherein in the preparation method of the water extract of the malva seed, the periodic cycle of the extraction is preferably carried out according to ultrasonic extraction for 6min and stirring extraction for 15 min; the extraction time is preferably 140-160min.
6. The extract of the fractional enrichment of the malva seed according to claim 4, wherein in the preparation method of the water extract of the malva seed, a pH regulator is used for regulating the pH, wherein the pH regulator is citric acid, and the pH is preferably 5.5-6.5.
7. The extract of the fractional enrichment of the malva seed according to claim 1, wherein the preparation method of the oil extract of the malva seed comprises the following steps: the method comprises the steps of (1 g) mixing the extract of the malva seed according to the feed liquid ratio of 1g: mixing 15-20 mL of petroleum ether, extracting at 40-70 ℃ for 60-120 min under stirring, separating solid from liquid after extraction, drying the leaching liquor to obtain the malva seed oil extract, wherein the filter residue is the malva seed extract powder 2.
8. The extract of the fractional enrichment of the malva seed according to claim 1, wherein the preparation method of the ethanol extract of the malva seed comprises the following steps: the method comprises the steps of mixing the extract powder 2 of the malva seed according to a feed liquid ratio of 1g: mixing 20-25 mL of ethanol water solution, extracting at 70-90 ℃ for 2-7min according to ultrasonic extraction, stirring and extracting for 10-15min for a period of cyclic extraction, extracting for 90-150min, carrying out solid-liquid separation after extraction, and drying the leaching liquor to obtain the ethanol extract of the malva seed.
9. The extract of the fractional enrichment of the malva seed according to claim 8, wherein in the preparation method of the ethanol extract of the malva seed, the ethanol aqueous solution is 20% -40% ethanol aqueous solution.
10. Use of the fraction enriched extract of malva seed according to any one of claims 1 to 9 for the preparation of a medicament against respiratory diseases.
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