CN113209155A - Total alkaloid extract of lysimachia christinae hance and application thereof - Google Patents
Total alkaloid extract of lysimachia christinae hance and application thereof Download PDFInfo
- Publication number
- CN113209155A CN113209155A CN202110480959.XA CN202110480959A CN113209155A CN 113209155 A CN113209155 A CN 113209155A CN 202110480959 A CN202110480959 A CN 202110480959A CN 113209155 A CN113209155 A CN 113209155A
- Authority
- CN
- China
- Prior art keywords
- total alkaloid
- alkaloid extract
- extract
- lysimachia christinae
- desmodium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- 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)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- 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
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
-
- 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
- A61K2236/333—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
- 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
- A61K2236/39—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
- A61K2236/55—Liquid-liquid separation; Phase separation
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Mycology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Gastroenterology & Hepatology (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Epidemiology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a lysimachia christinae total alkaloid extract and a detection method and application thereof in preparing anti-hepatic fibrosis drugs. The desmodium total alkaloid extract is prepared by crushing, extracting with alcohol or water, recovering ethanol, concentrating, eluting by macroporous adsorption resin column chromatography (ethanol-water system), concentrating, and drying under reduced pressure, and the preparation method of the desmodium total alkaloid extract is reasonable and stable, has short production period, and is suitable for industrial production; the method adopts the thin-layer chromatography to identify the desmodium total alkaloid extract, adopts the ultra-high performance liquid chromatography-high resolution mass spectrometry to identify the main chemical components of the desmodium total alkaloid extract, adopts the ultraviolet-visible spectrophotometry to determine the content of the total alkaloid in the desmodium total alkaloid extract, and has accurate and stable method and simple operation; has good effect as the anti-hepatic fibrosis medicine.
Description
Technical Field
The invention relates to a traditional Chinese medicine extract and application thereof, in particular to a lysimachia christinae total alkaloid extract and application thereof in resisting hepatic fibrosis.
Background
Herba Desmodii Styracifolii, herba Trifolii Pratentis, and herba Veronicae Formosanae of LeguminosaePhyllodium pulchellum (L.) the dried roots of Desv are distributed mainly in the areas of Guangxi, Guangdong, Jiangxi, etc. Has effects in promoting blood circulation, dispelling blood stasis, clearing away heat, and eliminating water. It is often used for costal pain, irregular menstruation and amenorrhea. The introduction of medical literature, herba lysimachiae can be used for treating cold, fever, sore throat, ulcerative gingivitis, rheumatalgia, edema, hepatosplenomegaly, swelling and pain from injuries, insect bite and other symptoms, at present, the chemical components of herba lysimachiae (titled: Phenolic constraints from the roots of phylogenetic pulchellum [ authors ] Zong Y, Zhong M, Li D M, et al, university of Girald university of medicine [ published ] Journal of auxiliary Natural Products Research, 2014 07, 741. Puff 746, 2014 07, St. Rochikuyama Miyabe [ 2014 ] Chunychikuan of Gen. (St. ex Pakishini et al, Church.) of liver fibrosis resisting ingredients Research [ Miyao ] Wang super, chime, Zhang Ying, etc., university of medicine [ 2014 ] Biyao, St.424, and Biotic Research [ sic ] of medicine [ sic ] Yidaqing, Zhento, and Biotic Research [ 424 ] of medicine [ sic ] 427, et al, medicine [ sic ] of Chinese medicine [ antibiotic ] Biyas ] of Chinese, Zhento, Xue, Xuanqian, Xue, Xuan, Xun, Xuan, etc. ], XuanPhyllodium pulchellum(L.) Desv [ authors ] Ya-Chu F, Shi-Jun Y, Zhong-Long G, et al, China oceanic university [ publication name ] Molecules, 2018(06): 1361. suppl 1375, [ subject name ] research on chemical components of lysimachia christinae [ authors ] Vandachu, Guo Zhonglong, Xinlan Ting, etc., China oceanic university [ publication name ] Chinese patent medicine, 2017(06): 1195. suppl 1198), quality evaluation [ subject name ] research on quality evaluation method of lysimachia christinae medicinal materials [ chime, Zhang Bao, Wang super, etc. ], Guangxi medical research institute [ publication name ] Chinese patent medicine, 2016. suppl (01): 130. suppl) (133) and pharmacological activity [ publication name ] of lysimachia christinae ] biological activity on collagen production factors of rat liver cells, Zongzi Sichuan nationality (Hunan: 14, Centan Sichuan nationality) research on collagen cell factors, 2017(03) 49-51, titled name, herba Desmodii Styracifolii alkaloid-human liver star stimulating acetaldehydeEffect of Toxicodendron, Zhang BaoJING, Wangsuper, etc., the Guangxi national institute of medicine [ Canning name ] modern application pharmacy in China, 2017(01):4-7, [ problem name ] Effect of Total alkaloid of Desmodium styracifolium on HSC-T6 cell collagen protein and cytokine mRNA expression [ Auricularia, Hunan culture, Yusheng, etc., the Guangxi national institute of medicine [ Canning name ] modern medical sanitation, 2017(24):3709-, Effects of cytokines [ authors ] huangjieling, chime, yunsmen, guangxi chinese medicine university [ journal ] china experimental and prescriptions, 2013 (13):283 and 286 (titled) influence of lysimachia christinae total alkaloids on serum interferon-gamma and liver histopathology in rats with hepatic fibrosis [ authors ] huanglin ruyi, chime, poplar brightening, etc., published by the institute of national medicine of the Guangxi province [ published name ] Chinese medical science and technology, 2006(02): 101-: 38 to 40 and the like are studied more deeply, the preparation and quality detection related problems of the total alkaloid extract of the lysimachia christinae hance are not reported in related researches.
With the development of pharmaceutical technology, the low toxicity and high efficiency of refined natural plant extracts are more and more favored by people. Research shows that the main effective components of the lysimachia christinae hance are alkaloid components. In addition, the medicinal part of the lysimachia christinae hance is root, and the application has the defects of heavy weight, difficult storage, inconvenient transportation and the like. The prepared total alkaloid extract of the lysimachia christinae hance can ensure the consistency with the main effects of the medicinal materials, can make up the defects, and has the effects of enhancing the effect, reducing the toxicity and improving the quality. Therefore, the research and development of a preparation and quality detection method of the lysimachia christinae total alkaloid extract is very necessary to ensure the quality of the extract, is beneficial to developing a new medicinal approach of the lysimachia christinae hance, and provides a scientific basis for fully utilizing medicinal material resources and improving the application value of the medicinal material resources.
Disclosure of Invention
The invention aims to provide a lysimachia christinae total alkaloid extract and application thereof in preparing anti-hepatic fibrosis drugs.
In order to achieve the purpose, the invention adopts the following technical scheme:
the invention discloses a lysimachia christinae total alkaloid extract and application thereof, which comprises one or more of the following preparation methods and applications.
1. Preparation of lysimachia christinae total alkaloid extract
1.1, preparing a lysimachia christinae hance medicinal material extracting solution: taking and crushing a dry lysimachia christinae hance medicinal material, adding 10-20 times of 60% -80% ethanol by volume according to the weight of the dry medicinal material, performing reflux extraction for 2-4 times for 1-2 hours each time, filtering, combining filtrates, recovering ethanol from the filtrates, performing reduced pressure concentration on the filtrate without alcohol smell, and concentrating the filtrate until the material-liquid ratio of the extract is 1: 10-1: 15; (the ethanol content below refers to the volume content);
1.2 treatment of macroporous resin: soaking macroporous resin (D101) in 95% ethanol for 24 h, packing with a wet method, washing with 95% ethanol solution until the eluate is mixed with water (volume ratio of 1: 3) and does not become white and turbid, and washing with water until no alcohol smell exists;
1.3 operating method: and (3) putting the extracting solution into a treated macroporous resin column at the speed of 1-3 BV/h for adsorption, eluting by using 1-2 BV water, discarding the eluent, eluting by using 50-70% ethanol with the amount of 3-5 BV at the speed of 3-5 BV/h, collecting ethanol liquid eluent, concentrating the eluent under reduced pressure to obtain an extract, and drying in vacuum at the temperature of 50 ℃ to obtain the lysimachia christinae hance total alkaloid extract.
The preparation method of the total alkaloid extract of the lysimachia christinae hance can also comprise the following steps: taking a dry lysimachia christinae hance medicinal material, crushing, adding 70% ethanol with 20 times volume content for refluxing for 1.5 hours for the first time, adding 70% ethanol with 15 times volume content for refluxing for 1.5 hours for the second time, adding 70% ethanol with 10 times volume content for refluxing for 1.5 hours for the third time, filtering, combining filtrates, recovering ethanol from the filtrate, carrying out reduced pressure concentration on the filtrate without alcohol smell, and concentrating the filtrate to obtain an lysimachia christinae hance extract with a material-to-liquid ratio of 1: 10; adsorbing the extractive solution with processed macroporous resin (XAD 1180 or XAD 1600N) column at 2 BV/h, eluting with 1 BV water, discarding eluate, eluting with 4BV 60% ethanol at 4BV/h, collecting ethanol eluate, concentrating the eluate under reduced pressure to obtain extract, and vacuum drying at 50 deg.C to obtain herba Desmodii Styracifolii total alkaloid extract.
The quality detection of the desmodium total alkaloid extract comprises the following steps:
(1) carrying out chromatographic comparison identification on the lysimachia christinae total alkaloid extract by adopting a 5-methoxy-N, N-dimethyltryptamine reference substance and a lysimachia christinae reference medicinal material;
(2) performing mass spectrum qualitative analysis on 5-methoxy-N, N-dimethyl tryptamine in the lysimachia christinae total alkaloid extract;
(3) the content of total alkaloids in the total alkaloids extract of lysimachia christinae hance is quantitatively analyzed by ultraviolet-visible spectrophotometry.
Firstly, a detection method of the total alkaloid extract of the lysimachia christinae hance is given out:
identification of total alkaloid extract of lysimachia christinae hance (thin-layer chromatography)
1. Preparation of control solutions: taking a proper amount of the 5-methoxy-N, N-dimethyltryptamine reference substance, adding methanol to prepare a solution containing 1 mg per 1ml, and filtering with a 0.45 mu m microporous filter membrane to obtain the product;
2. preparing a lysimachia christinae hance reference medicinal material solution: taking about 5 g of herba lysimachiae control medicinal material coarse powder, adding 15 times of 70% ethanol, performing reflux extraction for 3 times, each time for 1 hour, filtering, combining filtrates, recovering ethanol from the filtrate, dissolving the thick paste with methanol, and filtering with a 0.45 mu m microporous membrane to obtain the herba lysimachiae control medicinal material coarse powder;
preparation of a test solution: taking a proper amount of desmodium total alkaloid extract, adding methanol for dissolving, and filtering by a microporous filter membrane of 0.45 mu m to obtain the desmodium total alkaloid extract;
3. the determination method comprises the following steps: respectively sucking 5-10 μ l of each of a control solution, a lysimachia christinae control medicinal material solution and a test solution, respectively dropping on the same silica gel G thin-layer plate, presaturating for 10 min with dichloromethane-methanol-ammonia water (8: 1: 0.1) as a developing agent, taking out, drying in the air, spraying a diluted bismuth potassium iodide test solution for color development, inspecting under sunlight, and displaying the same orange-red spots on the chromatogram of the test solution at the positions corresponding to the chromatograms of the lysimachia christinae control medicinal material and the control solution.
Second, the main chemical components of the total alkaloid extract of lysimachia christinae hance are identified (high performance liquid chromatography-mass spectrometry)
1. Preparation of a test solution: dissolving appropriate amount of herba Desmodii Styracifolii total alkaloid extract with methanol to obtain solution containing about 1 mg of herba Desmodii Styracifolii total alkaloid extract per 1ml, and filtering with 0.22 μm microporous membrane to obtain the final product;
2. the instrument comprises the following steps: a Thermo UITiMate 3000 ultra-high performance liquid chromatograph, which is provided with an online degasser, a quaternary gradient pump, a column incubator and an automatic sample injector; a Thermo Q-active Focus high resolution mass spectrometer; a one-ten-thousandth balance;
3. chromatographic conditions are as follows: thermo Accucore C18A chromatography column (100 × 2.1 mm, 2.6 μm); the mobile phase is methanol (A) -water (B); gradient elution (0-10 min, 10% -90% A); the flow rate is 0.3 ml/min, and the column temperature is 20 ℃; sample introduction amount: 1 mu L of the solution;
4. mass spectrometry condition ion source: electrospray (HESI); detection mode: a positive ion mode; mass scan range: m/z is 100-1200; capillary temperature: 320 ℃; volume flow of sheath gas: 40 psi, assist gas volume flow: 10 psi; spraying voltage: 3.3 kV; temperature of probe warmer: 350 ℃; mass resolution: 70000; collision Energy (CE): 20, 40, 60 eV;
5. the determination method comprises the following steps: absorbing a sample solution, analyzing according to chromatographic and mass spectrum conditions under 2.2.3 and 2.2.4 items to respectively obtain a total ion flow chart (TIC) and a mass spectrum, performing data processing such as peak extraction, peak matching and the like by using Xcalibur software, performing pretreatment by using Compound Discover 2.3 software, and extracting the mass spectrum of a main component 5-methoxy-N, N-dimethyltryptamine according to an accurate mass number;
6. measurement results and analysis: MS (ESI) M/z 219.15, 174.09, 159.07, 143.07, HRMS [ M + H]+Mass number of excimer ion peak m/z 219.14906 and 5-methoxy-N, N-dimethyltryptamine (C)13H19ON2The theoretical value is as follows: 218.1414), molecular weight 218, secondary mass spectrum corresponding to 5-methoxyCracking rule of the base-N, N-dimethyltryptamine. The MS cleavage process is as follows: [ M + H ]]+Generates an excimer ion peak M/z 219.15, and generates [ M-45+ H ] after the cracking of the branched dimethylamino group]+The m/z 174.09 fragment, the methoxy group at position 5, is further cleaved to yield fragments of m/z159.07 and 143.07, respectively, according to the following cleavage formulae:
the total alkaloid extract comprises the following main chemical components: 5-methoxy-N, N-dimethyltryptamine.
Third, content determination of total alkaloid extract of lysimachia christinae Hance (ultraviolet-visible spectrophotometry)
1. Test solution: taking about 13mg of the total alkaloid extract of the lysimachia christinae hance, precisely weighing, placing in a 25ml measuring flask, adding an appropriate amount of methanol to dissolve and dilute to a scale, shaking uniformly, precisely weighing 1ml, placing in a 10ml measuring flask, adding an appropriate amount of methanol to dissolve and dilute to a scale, shaking uniformly, taking methanol as a blank, measuring an ultraviolet spectrum, and enabling a sample solution to have maximum absorption at a wavelength of 275 +/-1 nm.
2. Comparison products: the lysimachia christinae hance contains tryptamine, 5-methoxy-N, N-dimethyltryptamine, 5-hydroxy-N, N-dimethyltryptamine and other alkaloid components taking tryptamine as a structural parent nucleus, and the ultraviolet spectrum of the tryptamine has the maximum absorption at the wavelength of 275 +/-1 nm, so that the tryptamine is selected as a reference substance for measuring the content of total alkaloids;
3. preparation of control solutions: accurately weighing a proper amount of tryptamine reference substances, and adding methanol to prepare a solution containing 500 micrograms per 1ml to obtain the tryptamine reference substances;
4. preparation of a standard curve: accurately sucking 0.1 ml, 0.2 ml, 0.3 ml, 0.4 ml, 0.5 ml and 0.6 ml of tryptamine reference solution, respectively placing the tryptamine reference solution in a 10ml measuring flask, adding methanol to dilute the solution to a scale, shaking the solution uniformly, measuring absorbance at 275 +/-1 nm by using the methanol as a blank through an ultraviolet-visible spectrophotometry, and drawing a standard curve by using the concentration (mu g/ml) as a horizontal coordinate and the absorbance as a vertical coordinate; the regression equation is Y =0.02750X +0.01593, r = 0.9998;
5. the determination method comprises the following steps:weighing herba Desmodii Styracifolii total alkaloid extract about 13mg, precisely weighing, placing in 25ml measuring flask, adding appropriate amount of methanol to dissolve and dilute to scale, shaking, precisely weighing 1ml, placing in 10ml measuring flask, adding appropriate amount of methanol to dissolve and dilute to scale, shaking, measuring with methanol as blank, measuring by ultraviolet-visible spectrophotometry, measuring absorbance, calculating alkaloid content, and using tryptamine (C) as the content of herba Desmodii Styracifolii total alkaloid extract10H12N2) Calculated to be not less than 50.0%.
Tryptamine (C) extracted from total alkaloids of lysimachia christinae10H12N2) The application of the compound in preparing anti-hepatic fibrosis drugs comprises the following steps:
1. model establishment and grouping: SPF-level female Wistar white rats with weight of 180 +/-12 g are bred in an environment with temperature and humidity controlled for 12 hours, are molded by adopting a method of injecting pig serum into an abdominal cavity without diet limitation, and are randomly divided into a model control group, a high-dose group (30 mg/kg) of the total alkaloids of the lysimachia christinae, a medium-dose group (20 mg/kg) of the total alkaloids of the lysimachia christinae, a low-dose group (10 mg/kg) of the total alkaloids of the lysimachia christinae, and a colchicine drug control group (0.1 mg/kg).
2. The test method comprises the following steps: the rats in the normal group and the model group were filled with equal volume of distilled water 1 time/day. After 8 weeks, all experimental animals were fasted for 1 night, anesthetized by intraperitoneal injection of pentobarbital sodium (40 mg/kg), then sacrificed by cervical dislocation, the right lobe of the liver was rapidly taken out, fixed by 40 g/L formaldehyde, embedded in paraffin and stained by HE, and observed under a light microscope.
3. The determination method comprises the following steps: and (3) applying a CMIAS color medical image analyzer, randomly selecting 10 visual fields for measurement on liver slices of rats in each group, calculating the ratio of the positive staining area to the liver visual field area, and inspecting the expression degrees of the types I, III and IV collagen. Test results show that the lysimachia christinae total alkaloids have good anti-hepatic fibrosis effect by inhibiting the synthetic expression of collagen I, collagen III and collagen IV and TGF-beta 1 of liver fibrosis rats.
Compared with the prior art, the invention has the prominent substantive characteristics and remarkable progress that:
1. the invention is prepared from lysimachia christinae hance for the first timeThe content of the total alkaloid tryptamine (C) is obtained10H12N2) More than 50 percent of lysimachia christinae total alkaloid extract and a quality detection method thereof are established.
2. The method adopts the thin-layer chromatography to identify the total alkaloid extract of the lysimachia christinae hance, is simple and easy to implement, and can meet the requirement of industrial production.
3. The method adopts a high performance liquid chromatography-mass spectrometry combined method to identify the main chemical components in the lysimachia christinae total alkaloid extract, and has the advantages of accuracy, reliability and strong specificity.
4. The invention adopts ultraviolet-visible spectrophotometry to directly measure the content of the total alkaloids in the lysimachia christinae hance, has accurate and simple method and can effectively control the quality of the total alkaloids extract in the lysimachia christinae hance.
5. The total alkaloid extract of the lysimachia christinae hance has certain purity, reduces impurity interference, can reduce adverse reaction risk and improve drug effect when used as a medicine and a raw material thereof, and has good application prospect.
6. According to the invention, through researching the desmodium total alkaloids, a preparation method and a quality detection method of a desmodium total alkaloid extract are established, which are beneficial to further development and utilization of related products of the desmodium, and for developing Guangxi special medicinal materials, products with high technology and high added value are developed, the market competitiveness is improved, and potential social benefits and economic benefits can be generated.
Drawings
FIG. 1 is a flow chart of a preparation process of the desmodium total alkaloid extract, and as can be seen from FIG. 1, the preparation method of the desmodium total alkaloid extract comprises the following steps: drying herba Desmodii Styracifolii, pulverizing, extracting with ethanol or water under reflux, concentrating, recovering ethanol, suspending the crude extract with water, subjecting to macroporous adsorbent resin column chromatography (ethanol-water) eluting, concentrating, and vacuum drying to obtain herba Desmodii Styracifolii total alkaloid extract;
FIG. 2 is a thin-layer chromatogram of the total alkaloid extract of lysimachia christinae Hance, from which it can be seen that the chromatogram of the total alkaloid extract of lysimachia christinae Hance, the chromatogram of the reference drug of lysimachia christinae Hance and the chromatogram of the reference drug of 5-methoxy-N, N-dimethyltryptamine show the same orange-red spots at the corresponding positions;
FIG. 3 is a primary mass spectrum of 5-methoxy-N, N-dimethyltryptamine as the main component in the total alkaloid extract of lysimachia christinae Hance, from which FIG. 3 the quasi-molecular ion peak M/z 219.14906[ M + H ] is shown]+According to the mass number (C) of the 5-methoxy-N, N-dimethyltryptamine13H19ON2The theoretical value is as follows: 218.1414);
FIG. 4 is a secondary mass spectrum of 5-methoxy-N, N-dimethyltryptamine as the main component in the total alkaloid extract of lysimachia christinae Hance, and it can be seen from FIG. 4 that the cracking rule of the cracking fragments in the secondary mass spectrum is consistent with that of 5-methoxy-N, N-dimethyltryptamine;
FIG. 5 is a diagram showing the ultraviolet absorption spectra of the total alkaloid extract of lysimachia christinae hance and the tryptamine reference substance of the present invention, and it can be seen from FIG. 5 that the tryptamine and the total alkaloid extract of lysimachia christinae hance both have the maximum absorption at 275 + -1 nm, and the absorption spectra are basically consistent.
Detailed Description
Preparing the desmodium total alkaloid extract:
example 1
Pulverizing dry herba Desmodii Styracifolii into coarse powder 1 kg, extracting with 20 kg volume of 70% ethanol under reflux for 3 times (each for 1.5 hr), filtering, mixing filtrates, recovering ethanol from the filtrate, vacuum concentrating, and concentrating until the material-liquid ratio is 1: 10; adsorbing the extract on a treated macroporous resin (D101) column at the speed of 2 BV/h, eluting with 1 BV of water, discarding the eluent, eluting with 4BV of 60% ethanol at the speed of 4BV/h, collecting the ethanol eluent, concentrating the eluent under reduced pressure to obtain an extract, and drying under vacuum at 50 ℃ to obtain the desmodium total alkaloid extract.
Example 2
Pulverizing dry herba Desmodii Styracifolii into coarse powder 0.5 kg, extracting with 5 kg volume of 65% ethanol under reflux for 4 times (each for 1 hr), filtering, mixing filtrates, recovering ethanol from the filtrate, vacuum concentrating, and concentrating to obtain herba Desmodii Styracifolii extract with a material-to-liquid ratio of 1: 15; adsorbing the extract on a treated macroporous resin (D101) column at the speed of 2 BV/h, eluting with 2 BV water, discarding the eluent, eluting with 4BV of 60% ethanol at the speed of 4BV/h, collecting the ethanol eluent, concentrating the eluent under reduced pressure to obtain an extract, and vacuum-drying at 50 ℃ to obtain the desmodium total alkaloid extract, carrying out quality detection on the prepared desmodium total alkaloid extract by adopting the quality detection method disclosed by the invention, detecting 5-methoxy-N, N-dimethyltryptamine, and measuring the content of the total alkaloids to be 50.14%.
Example 3
Crushing the dried lysimachia christinae hance medicinal materials into 0.5 kg of coarse powder, adding 70% ethanol with the volume content of 10 kg for reflux extraction for 1.5 hours for the first time, adding 8 kg of 70% ethanol for reflux extraction for 1.5 hours for the second time, adding 5 kg of 70% ethanol for reflux extraction for 1.5 hours for the third time, filtering, combining the filtrates, recovering ethanol from the filtrate, carrying out reduced pressure concentration for removing the ethanol smell, and concentrating to obtain the lysimachia christinae hance extract with the material-liquid ratio of 1: 10; adsorbing the extract on a treated macroporous resin (D101) column at the speed of 2 BV/h, eluting with 1 BV of water, discarding the eluent, eluting with 4BV of 60% ethanol at the speed of 4BV/h, collecting the ethanol eluent, concentrating the eluent under reduced pressure to obtain an extract, and vacuum-drying at 50 ℃ to obtain the desmodium total alkaloid extract, carrying out quality detection on the prepared desmodium total alkaloid extract by adopting the quality detection method disclosed by the invention, detecting 5-methoxy-N, N-dimethyltryptamine, and measuring the content of the total alkaloids to be 50.90%.
Example 4
Pulverizing dry herba Desmodii Styracifolii into coarse powder 0.5 kg, extracting with 10 kg volume of 80% ethanol under reflux for 3 times (each for 2 hr), filtering, mixing filtrates, recovering ethanol from the filtrate, concentrating under reduced pressure until the ethanol smell disappears, and concentrating to obtain herba Desmodii Styracifolii extract with a material-to-liquid ratio of 1: 10; adsorbing the above extractive solution with processed macroporous resin (XAD 1600N) column at speed of 2 BV/h, eluting with 1 BV water, discarding eluate, eluting with 4BV 60% ethanol at speed of 4BV/h, collecting ethanol eluate, concentrating the eluate under reduced pressure to obtain extract, and vacuum drying at 50 deg.C to obtain herba Desmodii Styracifolii total alkaloid extract, detecting quality of the prepared herba Desmodii Styracifolii total alkaloid extract with 5-methoxy-N, N-dimethyltryptamine, and determining total alkaloid content to be 51.11%.
Example 5:
pulverizing dry herba Desmodii Styracifolii into coarse powder 0.5 kg, extracting with 10 kg volume of 70% ethanol under reflux for 3 times (each for 1.5 hr), filtering, mixing filtrates, recovering ethanol from the filtrate, concentrating under reduced pressure until the ethanol smell disappears, and concentrating to obtain herba Desmodii Styracifolii extract with a material-to-liquid ratio of 1: 10; adsorbing the extract with treated macroporous resin (XAD 1180) column at 2 BV/h, eluting with 1 BV water, discarding the eluate, eluting with 4BV 60% ethanol at 4BV/h, collecting the ethanol eluate, concentrating the eluate under reduced pressure to obtain extract, and vacuum drying at 50 deg.C to obtain herba Desmodii Styracifolii total alkaloid extract, detecting the quality of the prepared herba Desmodii Styracifolii total alkaloid extract with 5-methoxy-N, N-dimethyltryptamine, and determining the total alkaloid content to be 50.21%.
Example 6 (quality test method of Total Alkaloids extract of Desmodium styracifolium)
1. Identification of total alkaloid extract of lysimachia christinae hance (thin-layer chromatography)
1.1 preparation of control solutions: taking a proper amount of the 5-methoxy-N, N-dimethyltryptamine reference substance, adding methanol to prepare a solution containing 1 mg per 1ml, and filtering with a 0.45 mu m microporous filter membrane to obtain the product;
1.2 preparation of herba lysimachiae contrast medicinal material solution: taking 5 g of lysimachia christinae hance crude powder, adding 15 times of 70% ethanol for reflux extraction for 3 times, 1 hour each time, filtering, combining filtrates, recovering ethanol from the filtrate, dissolving the thick paste with methanol, and filtering with a 0.45 mu m microporous membrane to obtain the extract;
1.3 preparation of test solution: taking a proper amount of desmodium total alkaloid extract, adding methanol for dissolving, and filtering by a microporous filter membrane of 0.45 mu m to obtain the desmodium total alkaloid extract;
1.4 assay: respectively sucking 5-10 μ l of each of a reference solution, a lysimachia christinae hance medicinal material reference solution and a sample solution, respectively dropping on the same silica gel G thin-layer plate, presaturating for 10 min by taking dichloromethane-methanol-ammonia water (8: 1: 0.1) as a developing agent, taking out, drying in the air, spraying a diluted bismuth potassium iodide test solution for color development, inspecting under sunlight, and displaying orange red spots on the sample chromatogram at the positions corresponding to the lysimachia christinae hance medicinal material and the reference substance chromatogram.
2. Content determination of total alkaloid extract of lysimachia christinae Hance (ultraviolet-visible spectrophotometry)
2.1 test solution: taking about 13mg of the total alkaloid extract of the lysimachia christinae hance, precisely weighing, placing in a 25ml measuring flask, adding an appropriate amount of methanol to dissolve and dilute to a scale, shaking uniformly, precisely weighing 1ml, placing in a 10ml measuring flask, adding an appropriate amount of methanol to dissolve and dilute to a scale, shaking uniformly, taking methanol as a blank, measuring an ultraviolet spectrum, and enabling a sample solution to have maximum absorption at a wavelength of 275 +/-1 nm.
2.2 selection of control: the existing research shows that the lysimachia christinae hance contains tryptamine, 5-methoxy-N, N-dimethyltryptamine, 5-hydroxy-N, N-dimethyltryptamine and other alkaloid components which take tryptamine as a structural parent nucleus, and the ultraviolet spectrum of the tryptamine has the maximum absorption at the wavelength of 275 +/-1 nm, so that the tryptamine is selected as a reference substance for measuring the content of total alkaloids;
2.3 preparation of control solutions: accurately weighing a proper amount of tryptamine reference substances, and adding methanol to prepare a solution containing 503 mug per 1ml to obtain the tryptamine;
2.4 preparation of Standard Curve: accurately sucking 0.1 ml, 0.2 ml, 0.3 ml, 0.4 ml, 0.5 ml and 0.6 ml of tryptamine reference substance solution, respectively placing the tryptamine reference substance solution in a 10ml measuring flask, adding methanol to dilute the solution to a scale, shaking the solution uniformly, measuring absorbance at 275 +/-1 nm by using methanol as a blank through an ultraviolet-visible spectrophotometry, drawing a standard curve by using the concentration mu g/ml as a horizontal coordinate and the absorbance as a vertical coordinate, wherein the regression equation is Y =0.02750X +0.01593, and r = 0.9998;
2.5 assay: weighing herba Desmodii Styracifolii total alkaloid extract about 13mg, precisely weighing, placing in 25ml measuring flask, adding appropriate amount of methanol to dissolve and dilute to scale, shaking, precisely weighing 1ml, placing in 10ml measuring flask, adding appropriate amount of methanol to dissolve and dilute to scale, shaking, measuring with methanol as blank by ultraviolet-visible spectrophotometry, measuring absorbance at 275 + -1 nm wavelength, calculating alkaloid content, and determining the total alkaloid content with tryptamine C10H12N2Not to calculate lessAt 50.0%.
The assay methodology was tested as follows:
1. preparation of a standard curve: 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, 0.5 mL and 0.6 mL of the tryptamine control solution are precisely sucked and respectively placed in a 10mL measuring flask, methanol is added to dilute the solution to a scale, the solution is shaken up, the methanol is used as a blank, the absorbance is measured at 275 +/-1 nm by an ultraviolet-visible spectrophotometry, the concentration mu g/mL is used as a horizontal coordinate, the absorbance is used as a vertical coordinate to draw a standard curve, the regression equation is Y =0.02750X +0.01593, and r =0.9998, which shows that the linear relation of tryptamine is good within the range of 5-30 mu g/mL.
2. And (3) precision test: the same test solution was continuously measured 6 times, and the RSD was 0.09%.
3. And (3) stability test: taking the same sample solution, and measuring at 0, 30, 45, 60, 90, and 120 min respectively to obtain the result
RSD is 1.1%, which indicates that the test solution is stable within 120 min.
4. And (3) repeatability test: taking the same batch of samples to prepare 6 parts of test solution, and determining according to a proposed method, wherein the result RSD is
1.01 %。
5. Recovery rate test: taking the known content of the total alkaloid extract of the lysimachia christinae hance, respectively and precisely adding a proper amount of tryptamine reference substances, measuring the content according to a proposed method, and calculating the recovery rate, wherein the average recovery rate is 95.16%, and the RSD is 2.3%.
The result of methodology test shows that the method is accurate, stable and simple to operate, and can ensure the quality of the product.
Example 7 (qualitative identification of major chemical components of Total Alkaloids extract of Desmodium styracifolium)
1. Preparation of a test solution: dissolving appropriate amount of herba Desmodii Styracifolii total alkaloid extract with methanol to obtain solution containing about 1 mg of herba Desmodii Styracifolii total alkaloid extract per 1ml, and filtering with 0.22 μm microporous membrane to obtain the final product;
2. the instrument comprises the following steps: a Thermo UITiMate 3000 ultra-high performance liquid chromatograph, which is provided with an online degasser, a quaternary gradient pump, a column incubator and an automatic sample injector; a Thermo Q-active Focus high resolution mass spectrometer; a one-ten-thousandth balance;
3. chromatographic conditions are as follows: thermo Accucore C18 chromatography column (100X 2.1 mm, 2.6 μm); the mobile phase is methanol (A) -water (B); gradient elution (0-10 min, 10% -90% A); the flow rate is 0.3 ml/min, and the column temperature is 20 ℃; sample introduction amount: 1 mu L of the solution;
4. mass spectrum conditions: an ion source: electrospray (HESI); detection mode: a positive ion mode; mass scan range: m/z is 100-1200; the capillary temperature was 320 ℃; the volumetric flow of the sheath gas is 40 psi, and the volumetric flow of the auxiliary gas is 10 psi; the spray voltage is 3.3 kV; the temperature of the probe heater is 350 ℃; mass resolution 70000; collision Energy (CE): 20, 40, 60 eV;
5. the determination method comprises the following steps: absorbing a sample solution, analyzing according to chromatographic and mass spectrum conditions under 2.2.3 and 2.2.4 items to respectively obtain a total ion flow chart (TIC) and a mass spectrum, performing data processing such as peak extraction, peak matching and the like by using Xcalibur software, performing pretreatment by using Compound Discover 2.3 software, and extracting the mass spectrum of a main component 5-methoxy-N, N-dimethyltryptamine according to an accurate mass number;
6. measurement results and analysis: MS (ESI) M/z 219.15, 174.09, 159.07, 143.07, HRMS [ M + H]+Excimer peak m/z 219.14906 and mass number of 5-methoxy-N, N-dimethyltryptamine (C)13H19ON2The theoretical value is as follows: 218.1414), the molecular weight is 218, the secondary mass spectrum conforms to the cracking rule of 5-methoxy-N, N-dimethyltryptamine. The MS cleavage process is as follows: [ M + H ]]+Generates an excimer ion peak M/z 219.15, and generates [ M-45+ H ] after the cracking of the branched dimethylamino group]+The m/z 174.09 fragment, the methoxy group at position 5, is further cleaved to yield fragments of m/z159.07 and 143.07, respectively, according to the following cleavage formulae:
the total alkaloid extract comprises the following main chemical components: 5-methoxy-N, N-dimethyltryptamine.
Example 8 anti-hepatic fibrosis action of Total Alkaloids extract of Desmodium styracifolium
The invention discloses a pharmacodynamic test of the total alkaloid extract of lysimachia christinae hance, which has the following results:
1. model establishment and grouping: SPF-level female Wistar white rats with weight of 180 +/-12 g are bred in an environment with temperature and humidity controlled for 12 hours, are molded by adopting a method of injecting pig serum into an abdominal cavity without diet limitation, and are randomly divided into a model control group, a high-dose group (30 mg/kg) of the total alkaloids of the lysimachia christinae, a medium-dose group (20 mg/kg) of the total alkaloids of the lysimachia christinae, a low-dose group (10 mg/kg) of the total alkaloids of the lysimachia christinae, and a colchicine drug control group (0.1 mg/kg).
2. The test method comprises the following steps: the rats in the normal group and the model group were filled with equal volume of distilled water 1 time/day. After 8 weeks, all experimental animals were fasted for 1 night, anesthetized by intraperitoneal injection of pentobarbital sodium (40 mg/kg), then sacrificed by cervical dislocation, the right lobe of the liver was rapidly taken out, fixed by 40 g/L formaldehyde, embedded in paraffin and stained by HE, and observed under a light microscope.
3. Index measurement: and (3) applying a CMIAS color medical image analyzer, randomly selecting 10 visual fields for measurement on liver slices of rats in each group, calculating the ratio of the positive staining area to the liver visual field area, and inspecting the expression degrees of the types I, III and IV collagen.
4. As a result: the types of collagen I, III and IV in the livers of rats in the high, medium and low dose groups of the lysimachia christinae hance total alkaloids and the drug control group are all obviously reduced, and the fiber interval is thinner. The results of immunohistochemistry on collagen were subjected to image analysis, and subjected to computer image gray scale scanning digital conversion, and the measured values of the areal densities of collagen types I, III, and IV (collagen area/liver tissue visual field area 100%) in each group are shown in Table 1. Test results show that the lysimachia christinae total alkaloids have good anti-hepatic fibrosis effect by inhibiting the synthetic expression of collagen I, collagen III and collagen IV and TGF-beta 1 of liver fibrosis rats.
In conclusion, the desmodium total alkaloid extract is prepared by adopting macroporous adsorption resin column chromatography, the desmodium total alkaloid extract is qualitatively identified by adopting thin-layer chromatography, the main chemical components of the desmodium total alkaloid extract are qualitatively identified by adopting an ultra-high performance liquid chromatography-high resolution mass spectrometry combined method, and the content of total alkaloid in the desmodium total alkaloid extract is measured by adopting an ultraviolet-visible spectrophotometry, so that the method is accurate and stable, is simple to operate, has low analysis and test cost, can meet the requirement of industrial production, and can effectively control the quality of the desmodium total alkaloid extract. The desmodium total alkaloid extract has definite anti-hepatic fibrosis effect and has better application prospect.
Claims (7)
1. A preparation method of a lysimachia christinae total alkaloid extract is characterized in that: the preparation method of the desmodium total alkaloid extract comprises the following steps:
(1) preparing a lysimachia christinae hance extract: taking lysimachia christinae hance powder, carrying out reflux extraction for 2-5 times by 15-20 times of ethanol with the volume content of 60% -80%, 1-3 h each time, combining filtrates, carrying out reduced pressure evaporation until no alcohol smell exists, carrying out volume fixing by using distilled water until the material-liquid ratio is 1: 10-1: 15, and uniformly shaking to obtain the lysimachia christinae hance powder;
(2) preparing a dried matter of the total alkaloid extract of the lysimachia christinae hance: and (3) taking the treated macroporous resin, loading the lysimachia christinae hance extract at the flow rate of 1-3 BV/h, removing impurities by using 1-2 BV of distilled water, eluting by using 50% -70% ethanol with the amount of 3-5 BV at the flow rate of 3-5 BV/h, collecting eluent, concentrating and drying to obtain the lysimachia christinae hance extract.
2. The desmodium total alkaloid extract of claim 1, wherein: the herba Desmodii Styracifolii total alkaloid extract contains 5-methoxy-N, N-dimethyl tryptamine (tryptamine C)10H12N2) Calculated to be not less than 50.0%.
3. The quality detection method of the desmodium total alkaloid extract of claim 2, which is characterized in that: the quality detection of the desmodium total alkaloid extract comprises the following steps:
(1) carrying out chromatographic comparison identification on the lysimachia christinae total alkaloid extract by adopting a 5-methoxy-N, N-dimethyltryptamine reference substance and a lysimachia christinae reference medicinal material;
(2) performing mass spectrum qualitative analysis on 5-methoxy-N, N-dimethyl tryptamine in the lysimachia christinae total alkaloid extract;
(3) the content of total alkaloids in the total alkaloids extract of lysimachia christinae hance is quantitatively analyzed by ultraviolet-visible spectrophotometry.
4. The quality detection method of the desmodium total alkaloid extract of claim 3, which is characterized in that: the identification of the total alkaloid extract of the lysimachia christinae hance is carried out by adopting the following steps:
(1) preparation of control solutions: taking a proper amount of the 5-methoxy-N, N-dimethyltryptamine reference substance, adding methanol to prepare a solution containing 1 mg per 1ml, and filtering with a 0.45 mu m microporous filter membrane to obtain the product;
preparing a lysimachia christinae hance reference medicinal material solution: taking 5 g of desmodium styracifolium crude powder, adding 15 times of 70% ethanol for reflux extraction for 2 times, 1 hour each time, filtering, combining filtrates, recovering ethanol from the filtrate, dissolving the thick paste with methanol, and filtering with a 0.45 mu m microporous membrane to obtain a desmodium styracifolium control medicinal material solution;
preparation of a test solution: taking a proper amount of desmodium total alkaloid extract, adding methanol for dissolving, and filtering by a microporous filter membrane of 0.45 mu m to obtain the desmodium total alkaloid extract;
(2) and (3) determination: respectively sucking 5-10 μ l of each of a control solution, a lysimachia christinae control medicinal material solution and a test solution, respectively dropping on the same silica gel G thin-layer plate, presaturating for 10 min with dichloromethane-methanol-ammonia water (8: 1: 0.1) as a developing agent, developing, taking out, drying, spraying a diluted bismuth potassium iodide test solution for color development, inspecting under sunlight, and displaying orange red spots on the positions corresponding to the chromatograms of the lysimachia christinae medicinal material and the control in the chromatogram of the test solution.
5. The quality detection method of the desmodium total alkaloid extract of claim 3, which is characterized in that: the qualitative analysis of the 5-methoxy-N, N-dimethyl tryptamine in the desmodium total alkaloid extract is carried out by adopting the following steps:
(1) preparation of a test solution: dissolving appropriate amount of herba Desmodii Styracifolii total alkaloid extract with methanol to obtain solution containing about 0.3 mg of herba Desmodii Styracifolii total alkaloid extract per 1ml, and filtering with 0.22 μm microporous membrane to obtain the final product;
(2) the instrument comprises the following steps: an ultra-high performance liquid chromatography-high resolution mass spectrometer;
chromatographic conditions are as follows: c18 chromatography column (100X 2.1 mm, 2.6 μm); the mobile phase is methanol (A) -water (B); gradient elution (0-10 min, 10% -90% A); the flow rate is 0.3 ml/min; the column temperature is 20 ℃; sample introduction amount: 1 mu L of the solution;
mass spectrometry condition ion source: electrospray (HESI); detection mode: a positive ion mode; mass scan range: m/z is 100-1200; capillary temperature: 320 ℃; volume flow of sheath gas: 40 psi, assist gas volume flow: 10 psi; spraying voltage: 3.3 kV; temperature of probe warmer: 350 ℃; the mass resolution is more than or equal to 70000; collision Energy (CE): 20, 40, 60 eV;
(3) the determination method comprises the following steps: absorbing a sample solution, analyzing according to chromatographic and mass spectrum conditions under items 2.2.3 and 2.2.4 to respectively obtain a total ion flow chart (TIC) and a mass spectrum, performing data processing such as peak extraction, peak matching and the like by using Xcalibur software, performing pretreatment by using Compound Discover 2.3 software, and extracting a main component 5-methoxy-N, N-dimethyltryptamine (C) with accurate mass number13H19ON2The theoretical value is as follows: 218.1414) mass spectrum, MS (ESI) excimer ion [ M + H]+m/z 219, molecular weight 218, characteristic fragment ions m/z 174, 159 and 143, secondary mass spectrum conforms to the cracking rule of 5-methoxy-N, N-dimethyltryptamine, and the main chemical components are as follows: 5-methoxy-N, N-dimethyltryptamine.
6. The quality detection method of the desmodium total alkaloid extract of claim 3, which is characterized in that: the method is characterized in that: the content of total alkaloids is quantitatively analyzed by ultraviolet-visible spectrophotometry by the following steps:
(1) comparison products: the lysimachia christinae hance contains tryptamine and 5-methoxy-N, N-dimethyltryptamine, N-dimethyltryptamine and 5-hydroxy-N, N-dimethyltryptamine which take tryptamine as alkaloid components of a structural mother nucleus, so that the tryptamine is selected as a reference substance for measuring the content of total alkaloids;
preparation of control solutions: accurately weighing a proper amount of tryptamine reference substances, and adding methanol to prepare a solution containing 500 micrograms per 1ml to obtain the tryptamine reference substances;
(2) preparation of a standard curve: accurately sucking 0.1 ml, 0.2 ml, 0.3 ml, 0.4 ml, 0.5 ml and 0.6 ml of tryptamine reference substance solution, respectively placing the tryptamine reference substance solution in a 10ml measuring flask, adding methanol to dilute the solution to a scale, shaking the solution uniformly, measuring absorbance at 275 +/-1 nm by using methanol as a blank through an ultraviolet-visible spectrophotometry, drawing a standard curve by using the concentration mu g/ml as a horizontal coordinate and the absorbance as a vertical coordinate, wherein the regression equation is Y =0.02750X +0.01593, and r = 0.9998;
(3) the determination method comprises the following steps: weighing herba Desmodii Styracifolii total alkaloid extract about 13mg, precisely weighing, placing in 25ml measuring flask, adding appropriate amount of methanol to dissolve and dilute to scale, shaking, precisely weighing 1ml, placing in 10ml measuring flask, adding appropriate amount of methanol to dissolve and dilute to scale, shaking, measuring with methanol as blank, measuring by ultraviolet-visible spectrophotometry, measuring absorbance, calculating alkaloid content, and using tryptamine (C) as total alkaloid content in herba Desmodii Styracifolii total alkaloid extract10H12N2) Calculated to be not less than 50.0%.
7. The use of the total alkaloid extract of lysimachia christinae hance as claimed in claims 1-2, wherein: application of herba Desmodii Styracifolii total alkaloid extract in preparing medicine for resisting hepatic fibrosis is provided.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110480959.XA CN113209155A (en) | 2021-04-30 | 2021-04-30 | Total alkaloid extract of lysimachia christinae hance and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110480959.XA CN113209155A (en) | 2021-04-30 | 2021-04-30 | Total alkaloid extract of lysimachia christinae hance and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113209155A true CN113209155A (en) | 2021-08-06 |
Family
ID=77090345
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110480959.XA Pending CN113209155A (en) | 2021-04-30 | 2021-04-30 | Total alkaloid extract of lysimachia christinae hance and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113209155A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116077550A (en) * | 2023-02-14 | 2023-05-09 | 柳州市渔业技术推广站 | Chinese herbal medicine preparation for stimulating fresh water snail shell-out activity and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101138634A (en) * | 2006-09-07 | 2008-03-12 | 于保法 | Composition for treating tumour |
| CN104435977A (en) * | 2014-11-18 | 2015-03-25 | 西双版纳傣族自治州民族医药研究所 | Medicine for treating liver injuries and preparation method of medicine |
| CN110141602A (en) * | 2019-05-20 | 2019-08-20 | 西南大学 | A kind of extracting method and application of Folium Mori alkaloid |
-
2021
- 2021-04-30 CN CN202110480959.XA patent/CN113209155A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101138634A (en) * | 2006-09-07 | 2008-03-12 | 于保法 | Composition for treating tumour |
| CN104435977A (en) * | 2014-11-18 | 2015-03-25 | 西双版纳傣族自治州民族医药研究所 | Medicine for treating liver injuries and preparation method of medicine |
| CN110141602A (en) * | 2019-05-20 | 2019-08-20 | 西南大学 | A kind of extracting method and application of Folium Mori alkaloid |
Non-Patent Citations (4)
| Title |
|---|
| 干宁，等: "毛排钱草的化学成分及其对肿瘤细胞的细胞毒活性研究", 《中国中药杂志》 * |
| 钟鸣，等: "排钱草总生物碱对免疫性肝纤维化大鼠Ⅰ、Ⅲ、Ⅳ型胶原及TGF-β1表达的影响", 《中西医结合肝病杂志》 * |
| 钟鸣，等: "排钱草药材质量评价方法的研究", 《中成药》 * |
| 陈少峰，等: "壮药排钱草总生物碱对大鼠肝星状细胞胶原和细胞因子生成的影响", 《中国民族医药杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116077550A (en) * | 2023-02-14 | 2023-05-09 | 柳州市渔业技术推广站 | Chinese herbal medicine preparation for stimulating fresh water snail shell-out activity and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109324126B (en) | Method for simultaneously determining 9 chemical components in spina date seeds by using UPLC-MS/MS | |
| CN105111255A (en) | Extraction and separation method for echinacoside and verbascoside in cistanche | |
| CN108008026B (en) | Method for synchronously detecting 13 colorants in hawthorn pills | |
| Tian et al. | Orthogonal test design for optimization of the extraction of polysaccharide from Paeonia sinjiangensis KY Pan | |
| Zhang et al. | Quality evaluation of traditional Chinese drug toad venom from different origins through a simultaneous determination of bufogenins and indole alkaloids by HPLC | |
| CN109655558B (en) | Method for detecting effective part group of periploca forrestii schltr | |
| CN113209155A (en) | Total alkaloid extract of lysimachia christinae hance and application thereof | |
| CN108037200B (en) | Quality detection method of kidney nourishing and tranquilizing pills | |
| CN1803827A (en) | Method for preparing plant steroid glucoside | |
| CN109374774B (en) | Content determination method for simultaneously determining 4 components in cassia seeds by HPLC isocratic elution | |
| CN111983120B (en) | Method for establishing characteristic map of wind-dispelling pill mother and measuring content of 7 nucleoside components | |
| CN104569190A (en) | Method for rapidly determining four tannin substances in sapium sebiferum leaves | |
| CN113640448A (en) | Quality control method and construction method of saffron crocus | |
| CN110687224B (en) | Method for measuring triptolide A in tripterygium wilfordii medicinal material and tripterygium wilfordii multi-glycoside tablet prepared from tripterygium wilfordii medicinal material | |
| CN109966476B (en) | Application of three components of scutellaria baicalensis to synergistic enhancement of FGF2 cell proliferation promotion | |
| CN111116700B (en) | Method for extracting, separating and purifying dioscin from chrysanthemum leaves | |
| CN111337583A (en) | Method for measuring content of index components in urine and separating and identifying 20 metabolites after oral administration of phyllanthus emblica tannin parts | |
| CN112782326A (en) | Method for simultaneously measuring fingerprint spectrum and multi-index component content of Weikangling capsule | |
| CN101596274A (en) | The method of quality control of Fructus Schisandrae Chinensis in the YIXINSHU Chinese medicine preparation | |
| CN111956734A (en) | Optimized extraction method of yankang buccal tablets | |
| CN104059111A (en) | Method for extracting manninotriose monomer from rehmannia by virtue of activated carbon gradient elution | |
| Li et al. | Simultaneous determination of p-hydroxybenzaldehyde, p-hydroxybenzyl alcohol, 4-(β-D-glucopyranosyloxy)-benzyl alcohol, and sugars in Gastrodia elata blume measured as their acetylated derivatives by GC-MS | |
| CN113201037B (en) | Compound and Xian Mao Biaozhun decoction containing same | |
| CN114720602B (en) | Method for constructing characteristic spectrum of lithospermum through medicinal material or standard decoction thereof and detection method | |
| CN116818918B (en) | Fingerprint construction method of ligustrum robustum and construction method of high-resolution mass spectrum component basic identification and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210806 |