CN115015534A - Method for screening active substances in momordica grosvenori and application - Google Patents
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8693—Models, e.g. prediction of retention times, method development and validation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
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- Biochemistry (AREA)
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Abstract
The invention discloses a method for screening active substances in momordica grosvenori, which comprises the following steps: s1, purchasing a pharmaceutical preparation; s2, constructing an animal model; s3, animal model verification: s31, H & E staining pathological examination; s32, ELISA test: measuring content changes of IgE, IL-5, IL-13 and IL-17A in serum by using an ELISA kit according to an instruction; s4, instrumental analysis and measurement: detecting the liver tissues of mice in groups C, M, V and SGFE which are obtained before by adopting liquid phase-mass spectrometry combined technology; s5, metabonomics analysis; the invention mainly focuses on the change of liver metabolites after diseases occur, and compares the activity of monomers and coarse substances and the action mechanism thereof by comparing the dry state of the coarse substances and the monomer substances and comparing the activity of the monomers and the coarse substances and the action mechanism thereof by combining pharmacodynamic results, so as to clarify the main active ingredients and basis thereof for relieving asthma pneumonia by fructus momordicae.
Description
Technical Field
The invention belongs to the technical field of pharmacy, and particularly relates to a method for screening active substances in momordica grosvenori and application thereof.
Background
The momordica grosvenori is a Chinese characteristic medicine for both food and medicine, and has high edible value and medicinal value. The phytochemical research result shows that more than 20 cucurbitane triterpenoids and a plurality of flavonoids exist in the momordica grosvenori. The mogroside compounds are reported as the main substances in the grosvenor momordica, wherein the mogroside V is used as a component with the efficacy of relieving cough and moistening lung, and the content of the mogroside V can reach 52 percent of the total saponins. The mogroside can reduce lung inflammation caused by OVA by reducing Th2 type cell specific immunoreaction and reducing factors such as multiple Interleukins (ILs), tumor necrosis factor (TNF-alpha), gamma-interferon (IFN-gamma) and the like, and can also treat acute lung injury caused by LPS by regulating NF-kB pathway and reducing the content of nitric oxide synthase (iNOS) and cyclooxygenase (COX-2).
Metabonomics is a global system biology research method, and utilizes modern analysis technology with high flux, high sensitivity and high accuracy to carry out dynamic tracking analysis on the whole composition of metabolites in body fluid and tissues, finds and searches biomarkers by means of a multivariate statistical analysis method, and identifies and analyzes the physiological and pathological states of a researched object and the relationship between the physiological and pathological states and the environmental factors and the genetic composition of the researched object to carry out disease diagnosis and pathogenesis research. Metabolomics is therefore an effective approach to study the mechanisms of action of drugs in interfering with disease. Metabolomics is divided into 2 groups, non-targeted metabolomics and targeted metabolomics. Non-targeted metabonomics do not separate and identify a single metabolite, but analyze endogenous small molecules of the whole body, and aims to find component differences among different groups of samples and confirm biomarkers; the targeted metabolome separates and utilizes an internal standard substance to identify a single component and analyze a specific small molecule or specific small molecules, and aims to analyze metabolic pathways related to the specific component so as to determine the pathological mechanism or the pharmacodynamic target of the disease. Essentially, genomics has the same function as transcriptomics and proteomics, which are both system biology, and can analyze the biological function of molecules. However, unlike the latter two, the metabolome is a downstream product of genome and proteome, and when the protein and gene changes to be small and impossible, the metabolomics can still amplify biological information and detect the change of metabolites due to the sensitivity thereof, thereby truly reflecting the function of a biological system.
The efficacy of momordica grosvenori in moistening lung and relieving cough is long-standing, and a plurality of basic researches show that momordica grosvenori relieves cough symptoms and reduces sputum secretion by relieving inflammation generated in the lung, but the momordica grosvenori is used in the initial stage of taking the momordica grosvenori by soaking in water for a long time due to unclear functional components. The administration mode makes the disease-relieving degree of the momordica grosvenori difficult to control and makes certain standards to measure the drug effect difficult. This obviously limits the important applications of luo han guo. Therefore, a method for screening the main active ingredients with the efficacy of moistening lung and relieving cough from the momordica grosvenori is needed at present. This approach needs to be mechanistically driven to find out whether it has a modulating effect on the improvement of pulmonary inflammation. However, due to the synergistic effect of multiple components and multiple targets, the research on the whole system from tissues, organs, cells and molecules is difficult, and the material basis of the core efficacy is not clear.
In recent years, the research of traditional Chinese medicine is more and more focused by people, and the modernization research process of traditional Chinese medicine is also continuously promoted. The active ingredients of traditional Chinese medicines are characterized by definite drug effect, various skeleton structures, convenient access, wide biological activity and the like, and the screening of active substances of medicines taking traditional Chinese medicines as sources is increasingly emphasized by new medicine researchers. The traditional Chinese medicine active ingredient screening research process has long period, large workload and low efficiency, and is easy to lose a few trace of potential active substances and be interfered by false positive results. The traditional screening method is based on the combination of screening methods such as serum medicinal chemistry, serum pharmacology, metabonomics method, molecular recognition technology and the like, three technologies of molecular docking, high-flux screening and cell membrane chromatography are developed in recent years, mainly focusing on medicines, and the technologies mainly aim at the medicine and the in-vivo metabolic change thereof and do not pay attention to the change of the metabolic substances of the organism.
The invention mainly focuses on the change of liver metabolites after diseases occur, and compares the activity of monomers and coarse substances and action mechanisms thereof by comparing the changes of the coarse substances and the monomer substances and combining pharmacodynamic results, and aims to clarify main active ingredients and basis of momordica grosvenori for relieving asthmatic pneumonia. Therefore, a screening method and application of active substances in momordica grosvenori are provided to solve the problems mentioned in the background technology.
Disclosure of Invention
The invention aims to provide a screening method and application of active substances in momordica grosvenori, so as to solve the problems in the background technology.
In order to achieve the purpose, the invention provides the following technical scheme: a method for screening active substances in fructus momordicae comprises the following steps:
s1, purchasing a pharmaceutical preparation:
purchasing mogroside V, crude mogroside extract, Suhuang cough relieving capsule, ovalbumin and Al (OH) 3 ；
S2, constructing an animal model:
45 female BALB/c mice were purchased from Spanish lake laboratory animals, Inc.;
group C: animals were divided into blank groups, M groups: model group, SGFE group: crude extract group of mogroside, group V: mogroside group V and group P: a positive drug group;
on the 0 th, 7 th and 14 th days, mice in M group, V group, SGFE group and P group are respectively given 0.1ml of sensitizing solution for intraperitoneal injection, and group C is given PBS solution for intraperitoneal injection;
spraying 1% OVA solution to mice of group M, group V, group SGFE and group P every day on 21-32 days, and respectively administering mogroside V50 mg/kg, mogroside crude extract 50mg/kg and Suhuang cough capsule 50mg/kg to mice of group V and group P;
killing the mice 24 hours after the last spraying, and taking lung, liver, alveolar lavage fluid and blood;
s3, animal model verification:
s31, H & E staining pathological examination;
s32, ELISA test:
measuring content changes of IgE, IL-5, IL-13 and IL-17A in serum by using an ELISA kit according to an instruction;
s4, instrumental analysis and measurement:
detecting the liver tissues of mice in groups C, M, V and SGFE which are obtained before by adopting liquid phase-mass spectrometry combined technology;
s5, metabolomics analysis:
introducing the annotated metabolites in the liver into SIMCA-P14.0 for principal component analysis, respectively performing orthogonal partial least squares analysis on a group C and a group M and a group V or an SGFE group, evaluating a model and screening out key differential metabolites; differential metabolites are enriched, and differential change pathways between Metabionalys 5.0M group and C group, M group and V group or SGFE group are introduced and enriched.
In the step S1, the mogroside V is purchased from Pusi Biotechnology Ltd, the purity of the mogroside V is not less than 95%, and the CAS number is 88901-364;
the crude extract of mogroside is obtained from Rhine corporation, and the purity of mogroside V is not less than 52.26%;
suhuang cough relieving capsules were purchased from Yangzhijiang pharmaceutical industries, Ltd;
the ovalbumin is purchased from leaf biotechnology limited, and the purity of the ovalbumin is 98 percent;
Al(OH) 3 from sienna herd ltd.
The instrument analysis and measurement in step S4 specifically includes:
s41, chromatographic conditions & mass spectrometric detection:
the chromatographic instrument adopts Thermo Ultimate 3000, an ACQUITY UPLC HST 31.8μm (2.1X 150 mm) chromatographic column is used, the temperature of an autosampler is set to be 8 ℃, gradient elution is carried out by feeding 2 microliter at the flow rate of 0.25mL/min and the column temperature of 40 ℃, and the mobile phase is 0.1% of formic acid water (C) -0.1% of formic acid acetonitrile (D) as positive ions; negative ion 5mM ammonium formate water (a) -acetonitrile (B);
the mass spectrometer uses Thermo Q active Plus, electrospray ionization (ESI) and positive and negative ion ionization modes, wherein the positive ion spray voltage is 4.20kV, the negative ion spray voltage is 3.50kV, the sheath gas is 30arb, and the auxiliary gas is 10 arb; the temperature of the capillary tube is 325 ℃, full scanning is carried out with the resolution of 70000, the scanning range is 81-1000, secondary cracking is carried out by adopting HCD, the collision voltage is 30eV, and unnecessary MS/MS information is removed by adopting dynamic exclusion;
s42, data processing:
identification of metabolites was first confirmed by accurate molecular weight with a molecular weight error of < =30ppm, and subsequently annotated by MS/MS fragmentation patterns to Human Metabolome Database (HMDB), METLIN, Massbank, lipedmaps and mzbound databases to obtain metabolites.
An application of a screening method of active substances in fructus momordicae is applied to lung inflammation induced by SOVA.
Compared with the prior art, the invention has the beneficial effects that: the invention provides a method for screening active substances in momordica grosvenori and application thereof, which mainly pay attention to the change of liver metabolites after diseases occur, compare the changes of body liver metabolites after the coarse body components and the monomer components are compared, compare the activities of the monomers and the coarse body components and the action mechanism thereof by combining pharmacodynamic results, and aim to clarify the main active components and basis of the momordica grosvenori for relieving asthma pneumonia.
The invention determines that the mogroside V is the main effective component of the crude extract by a metabolic pathway research and combination analysis method. Effective active ingredients in the momordica grosvenori are innovatively found out through the research method, and a new search idea can be provided for searching for active compounds taking effect in traditional Chinese medicine extraction mixtures. The mogroside V is obtained by comparing the metabolic pathway mechanism of natural active ingredients and is an effective active ingredient. The research method can provide a new research idea for determining the effective components in the natural traditional Chinese medicine compounds in the future.
Drawings
FIG. 1 is a schematic flow chart of the method for screening active substances in Siraitia grosvenorii according to the present invention;
FIG. 2 is a schematic diagram of a regulation mechanism network according to the present invention;
FIG. 3 is a graph showing the change in the IgE content in serum according to the present invention;
FIG. 4 is a graph showing the change in the IL-5 content in serum according to the present invention;
FIG. 5 is a graph showing the change in the IL-13 content in serum according to the present invention;
FIG. 6 is a graph showing the change in the IL-17A content in serum according to the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention provides a method for screening active substances in momordica grosvenori shown in figures 1-6, which comprises the following steps:
s1, purchasing a pharmaceutical preparation:
purchasing mogroside V, crude mogroside extract, Suhuang cough relieving capsule, ovalbumin and Al (OH) 3 ；
Mogroside V is purchased from Pusi Biotechnology Ltd, the purity of the mogroside V is more than or equal to 95%, and the CAS number is 88901-;
the crude extract of mogroside is obtained from Rhine Ipomoea limited, and the purity of mogroside V is more than or equal to 52.26%);
suhuang cough relieving capsules were purchased from Yangzhijiang pharmaceutical industries, Ltd;
the ovalbumin is purchased from leaf biotechnology limited, and the purity of the ovalbumin is 98 percent;
Al(OH) 3 from sienna herd ltd;
s2, constructing an animal model:
45 female BALB/c mice were purchased from Spanish lake laboratory animals, Inc.;
group C: animals were divided into blank groups, M groups: model group, SGFE group: crude extract group of mogroside, group V: mogroside group V and group P: a positive drug group;
animals were divided into 5 groups of blank group (C, control), model group (M, model), mogroside crude extract group (SGFE, Siraitia grosvenorii front extract), mogroside V (mogeoside V), mogroside crude extract (SGFE), and positive drug (P, suhuang cough capsule). On the 0 th, 7 th and 14 th days, mice in M group, V group, SGFE group and P group are respectively subjected to intraperitoneal injection of 0.1ml of sensitizing solution, and mice in blank group are subjected to intraperitoneal injection of PBS solution;
spraying 1% OVA solution to mice of M group, V group, SGFE group and P group every day on 21-32 days, and respectively administering mogroside V group and P group mice V50 mg/kg, mogroside crude extract 50mg/kg and Suhuang cough relieving capsule 50 mg/kg;
mice were sacrificed 24 hours after the last spray, and lung, liver, alveolar lavage fluid (BALF) and blood were taken;
s3, animal model verification:
s31, H & E staining pathological examination;
s32, ELISA test:
measuring content changes of IgE, IL-5, IL-13 and IL-17A in serum by using an ELISA kit according to an instruction;
s4, instrumental analysis and measurement:
detecting the liver tissues of the mice in the groups C, M, V and SGFE obtained before by adopting a liquid phase-mass spectrometry combined technology (LC-MS);
chromatographic conditions & mass spectrometric detection:
the chromatographic instrument adopts Thermo Ultimate 3000, an ACQUITY UPLC HST 31.8μm (2.1X 150 mm) chromatographic column is used, the temperature of an autosampler is set to be 8 ℃, gradient elution is carried out by feeding 2 microliter at the flow rate of 0.25mL/min and the column temperature of 40 ℃, and the mobile phase is 0.1% of formic acid water (C) -0.1% of formic acid acetonitrile (D) as positive ions; negative ion 5mM ammonium formate water (A) -acetonitrile (B).
The mass spectrometer used Thermo Q active Plus, electrospray ion source (ESI), positive and negative ion ionization mode, positive ion spray voltage of 4.20kV, negative ion spray voltage of 3.50kV, sheath gas 30arb, and auxiliary gas 10 arb. Capillary temperature 325 ℃, full scan at resolution 70000, scan range 81-1000, and secondary lysis with HCD with collision voltage of 30eV, while removing unnecessary MS/MS information with dynamic exclusion.
Data processing:
identification of metabolites was first confirmed by accurate molecular weight (molecular weight error < =30 ppm), and subsequently annotated to databases such as Human Metamolome Database (HMDB), METLIN, Massbank, lipdmaps, and mzcloud according to MS/MS fragmentation patterns to obtain metabolites.
S5, metabolomics analysis:
the annotated metabolites in the liver were imported into SIMCA-P14.0 for Principal Component Analysis (PCA), orthogonal partial least squares analysis (OPLS-DA) was performed on group C and group M and group V or SGFE, respectively, the model was evaluated and key differential metabolites (VIP >1, P < 0.05) were screened. Differential metabolites are enriched, and differential change pathways between Metabionalys 5.0M group and C group, M group and V group or SGFE group are introduced and enriched.
An application of a screening method of active substances in momordica grosvenori is applied to lung inflammation induced by SOVA.
In the research, the grosvenor momordica saponin V and the crude saponin extract can improve OVA-induced lung inflammation, wherein the saponin V shows more excellent curative effect. Through the mechanism research of liver metabonomics, the mogroside V is obtained to improve the inflammation of the lung by regulating riboflavin metabolism (p =0.5), purine metabolism (p =0.05872) and glutathione metabolism (p = 0.0586); crude mogroside extracts regulate metabolism by riboflavin metabolism (p =0.5), phenylalanine metabolism (p =0.2619), glycerolipid metabolism (p =0.23676), sphingolipid metabolism (p =0.15416), glycine metabolism (p =0.23676), and phospholipid metabolism (p = 0.15416). Serine and threonine metabolism (p =0.08668), and arginine and proline metabolism (p =0.10868) to ameliorate lung inflammation.
By comparing their regulatory mechanism networks, we found that both are centered on riboflavin metabolism, with an impact index of 0.5, suggesting that riboflavin metabolism may be the primary regulatory pathway for regulating pulmonary inflammation. The content of the mogroside V in the crude extract is over half, but the curative effect is obviously better than that of the mogroside crude extract, which indicates that the saponin V is a main effective component playing a role in the saponin crude extract.
In conclusion, compared with the prior art, the invention aims to find the active ingredients in the momordica grosvenori which play the effects of moistening lung and relieving cough, and the momordica grosvenori saponin V and the crude extract (crude extract) of the momordica grosvenori saponin in the momordica grosvenori are respectively selected to explain the mechanism. Mogroside V accounts for about 52.26% of the crude extract, and both have been shown to have the effects of moistening lung and relieving cough. In our phenotypic studies, mogroside V has a more potent therapeutic effect than the crude extract. In the mechanism study, we found that the core study mechanism common to both is riboflavin metabolism and the influencing factor is 0.5. Riboflavin metabolism is a collective essential vitamin, usually acting in the form of Flavin Mononucleotide (FMN) and flavin adenine dinucleotide (FMN).
Riboflavin exerts immunomodulatory, antioxidant and anti-inflammatory effects in the body by participating in the metabolism of various macromolecules. The regulation of riboflavin metabolism is based on its ability to regulate energy metabolism. Inflammation is usually accompanied by the activation and differentiation of a large number of immune cells, which require a large amount of energy, and riboflavin metabolism can block the activation of macrophages, reduce immune response and reduce the secretion of inflammatory factors. Riboflavin also neutralizes free radicals such as active oxygen and H2O2 generated during respiration during glutathione metabolism. Other metabolic pathways regulated by both also include glutathione metabolism and purine metabolism.
But in addition to this, crude extracts also exert complex effects by regulating other metabolic pathways such as phenylalanine metabolism, glycerolipid metabolism, sphingolipid metabolism, porphyrin and chlorophyll metabolism, glycine, serine and threonine metabolism, arginine and proline metabolism. However, these metabolic pathways do not contribute significantly to the anti-inflammatory effect, which is consistent with our phenotypic studies. Therefore, the method of metabolic pathway research and combined analysis determines that the mogroside V is the main effective component of the crude extract. Effective active ingredients in the momordica grosvenori are innovatively found out through the research method, and a new search idea can be provided for searching for active compounds taking effect in traditional Chinese medicine extraction mixtures.
The mogroside V is obtained by comparing the metabolic pathway mechanism of natural active ingredients and is an effective active ingredient. The research method can provide a new research idea for determining the effective components in the natural traditional Chinese medicine compounds in the future.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments or portions thereof without departing from the spirit and scope of the invention.
Claims (4)
1. A method for screening active substances in momordica grosvenori is characterized in that: the method comprises the following steps:
s1, purchasing a pharmaceutical preparation:
purchasing mogroside V, crude mogroside extract, Suhuang cough relieving capsule, ovalbumin and Al (OH) 3 ；
S2, constructing an animal model:
45 female BALB/c mice were purchased from Spanish lake laboratory animals, Inc.;
group C: animals were divided into blank groups, M groups: model group, SGFE group: crude extract group of mogroside, group V: mogroside group V and group P: a positive drug group;
on days 0, 7 and 14, mice in groups M, V, SGFE and P are subjected to intraperitoneal injection of 0.1ml of sensitizing solution, and mice in group C are subjected to intraperitoneal injection of PBS solution;
spraying 1% OVA solution to mice of group M, group V, group SGFE and group P every day on 21-32 days, and respectively administering mogroside V50 mg/kg, mogroside crude extract 50mg/kg and Suhuang cough capsule 50mg/kg to mice of group V and group P;
killing the mice 24 hours after the last spraying, and taking lung, liver, alveolar lavage fluid and blood;
s3, animal model verification:
s31, H & E staining pathological examination;
s32, ELISA test:
measuring content changes of IgE, IL-5, IL-13 and IL-17A in serum by using an ELISA kit according to an instruction;
s4, instrumental analysis and measurement:
detecting the liver tissues of mice in groups C, M, V and SGFE which are obtained before by adopting liquid phase-mass spectrometry combined technology;
s5, metabolomics analysis:
introducing the annotated metabolites in the liver into SIMCA-P14.0 for principal component analysis, respectively performing orthogonal partial least squares analysis on a group C and a group M and a group V or an SGFE group, evaluating a model and screening out key differential metabolites; differential metabolites are enriched, and differential change pathways between Metabionalys 5.0M group and C group, M group and V group or SGFE group are introduced and enriched.
2. The method for screening active substances in siraitia grosvenorii as claimed in claim 1, wherein: in the step S1, the mogroside V is purchased from Pusi Biotechnology Ltd, the purity of the mogroside V is not less than 95%, and the CAS number is 88901-364;
the crude extract of mogroside is obtained from Rhine corporation, and the purity of mogroside V is not less than 52.26%;
suhuang cough relieving capsules were purchased from Yangzhijiang pharmaceutical industries, Ltd;
the ovalbumin is purchased from leaf biotechnology limited, and the purity of the ovalbumin is 98 percent;
Al(OH) 3 from sienna herd ltd.
3. The method for screening active substances in siraitia grosvenorii as claimed in claim 1, wherein: the instrument analysis and measurement in step S4 specifically includes:
s41, chromatographic conditions & mass spectrometric detection:
the chromatographic instrument adopts Thermo Ultimate 3000, ACQUITY UPLC HST 31.8μm and 2.1X 150mm chromatographic columns are used, the temperature of an autosampler is set to be 8 ℃, gradient elution is carried out by feeding 2 muL at the flow rate of 0.25mL/min and the column temperature of 40 ℃, and the mobile phase is positive ion 0.1% formic acid water (C) -0.1% formic acid acetonitrile (D); negative ion 5mM ammonium formate water (a) -acetonitrile (B);
the mass spectrometer uses Thermo Q active Plus, electrospray ion source ESI, positive and negative ion ionization mode, the positive ion spray voltage is 4.20kV, the negative ion spray voltage is 3.50kV, the sheath gas is 30arb, the auxiliary gas is 10 arb; the temperature of the capillary tube is 325 ℃, full scanning is carried out with the resolution of 70000, the scanning range is 81-1000, secondary cracking is carried out by adopting HCD, the collision voltage is 30eV, and unnecessary MS/MS information is removed by adopting dynamic exclusion;
s42, data processing:
identification of metabolites is firstly confirmed according to accurate molecular weight, the error of the molecular weight is < =30ppm, and then the identification is confirmed according to MS/MS fragment patterns on HMDB, METLIN, Massbank, LipidMaps and mzClound databases to obtain metabolites.
4. The application of the method for screening active substances in momordica grosvenori according to claim 1, wherein the method comprises the following steps: applied to lung inflammation induced by SOVA.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004038381A2 (en) * | 2002-10-25 | 2004-05-06 | Metabolon, Inc. | Methods for drug discovery, disease treatment, and diagnosis using metabolomics |
| KR20160033995A (en) * | 2014-09-19 | 2016-03-29 | 한국생명공학연구원 | Pharmaceutical composition for prevention or treatment of asthma comprising the extracts and fractions of Dendropanax morbifera as active ingredient |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004038381A2 (en) * | 2002-10-25 | 2004-05-06 | Metabolon, Inc. | Methods for drug discovery, disease treatment, and diagnosis using metabolomics |
| KR20160033995A (en) * | 2014-09-19 | 2016-03-29 | 한국생명공학연구원 | Pharmaceutical composition for prevention or treatment of asthma comprising the extracts and fractions of Dendropanax morbifera as active ingredient |
Non-Patent Citations (1)
| Title |
|---|
| YISA LIU: "The liver metabolic features of Mogroside V compared to Siraitia grosvenorii fruit extract in allergic pneumonia mice", 《MOLECULAR IMMUNOLOGY》, no. 145, 17 March 2022 (2022-03-17), pages 1 - 2 * |
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