CN110292588A - A kind of preparation method of Panax stipuleanatus Tsai et Feng total saposins and the application of anticoagulation medicine - Google Patents
A kind of preparation method of Panax stipuleanatus Tsai et Feng total saposins and the application of anticoagulation medicine Download PDFInfo
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- stipuleanatus tsai
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- 241000168721 Panax stipuleanatus Species 0.000 title claims abstract description 106
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- 238000002360 preparation method Methods 0.000 title claims abstract description 22
- 239000003814 drug Substances 0.000 title abstract description 11
- 230000010100 anticoagulation Effects 0.000 title abstract description 6
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- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims abstract description 42
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- 239000003146 anticoagulant agent Substances 0.000 claims abstract description 9
- 229940127219 anticoagulant drug Drugs 0.000 claims abstract description 8
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- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
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- 238000012449 Kunming mouse Methods 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 206010028813 Nausea Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241000208343 Panax Species 0.000 description 1
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- 206010047249 Venous thrombosis Diseases 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
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- OORMXZNMRWBSTK-LGFJJATJSA-N dammarane Chemical compound C1CCC(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@H]([C@H](C)CCCC(C)C)[C@H]4CC[C@@H]3[C@]21C OORMXZNMRWBSTK-LGFJJATJSA-N 0.000 description 1
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- 150000002632 lipids Chemical class 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
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- 230000001603 reducing effect Effects 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
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- 231100000331 toxic Toxicity 0.000 description 1
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- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 description 1
- 229940038773 trisodium citrate Drugs 0.000 description 1
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
- A61K36/258—Panax (ginseng)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
-
- 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
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
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- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
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- Public Health (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
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- Animal Behavior & Ethology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Alternative & Traditional Medicine (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
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- Diabetes (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
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Abstract
The present invention relates to the applications of a kind of preparation method of Panax stipuleanatus Tsai et Feng total saposins and anticoagulation medicine, belong to biomedicine technical field.Panax stipuleanatus Tsai et Feng powder is added in Extraction solvent by the present invention, 20 ~ 40 min of Microwave Extraction under the conditions of temperature is 60 ~ 100 DEG C, residue and extracting solution are obtained after filtering, residue is extracted 2 ~ 3 times under identical extraction conditions again, combined extract, it is concentrated under reduced pressure to give medicinal extract again, n-butanol medicinal extract is obtained using saturation extracting n-butyl alcohol medicinal extract, n-butanol medicinal extract uses MCI pillar layer separation, it carries out affording ethanol eluate with ethanol/water dicyandiamide solution, ethanol eluate is concentrated, is drying to obtain Panax stipuleanatus Tsai et Feng total saposins.Panax stipuleanatus Tsai et Feng total saposins of the present invention can be used for preparing treatment thrombotic diseases and/or anticoagulant pharmaceutical composition.
Description
Technical field
The present invention relates to the applications of a kind of preparation method of Panax stipuleanatus Tsai et Feng total saposins and anticoagulation medicine, belong to biological medicine
Technical field.
Background technique
Thrombus is fibrin and the deposition of shaped components in blood in the blood vessels.Anticoagulant factor and fibrinolysis system
The dysfunction of system can lead to the diseases such as apoplexy, thrombus and myocardial infarction.Currently, the case fatality rate of thrombus is only second to nauseous tumour
And myocardial infarction.Anticoagulation can inhibit the generation and fibrinous formation of fibrin ferment, to accelerate the dissolution of thrombus.On the market
It is larger for anticoagulant clinical medicine side effect, it is easy to induce the diseases such as haemorrhage, hyperkalemia, and natural anticoagulant comes
Source is abundant, and the characteristic with high-efficiency low-toxicity has huge potentiality in terms for the treatment of thrombotic diseases.
Panax stipuleanatus Tsai et Feng Panax stipuleanatus H.T.Tsai et K.M.Feng is Araliaceae Panax Panax
The rare medicinal herbs of linn, is used as medicine with root, have effects that strengthening the muscles and bones, stagnation resolvation cure the wound, analgesic therapy dredging collateral.Panax stipuleanatus Tsai et Feng also known as open country
Radix Notoginseng is the wild relatives of Radix Notoginseng, has significant difference both from mode of appearance.Radix Notoginseng main root is in short cone more, and is shielded
Side rhizomes of Panax notoginseng is crawled, tool recess stem trace.The main functional component of Panax stipuleanatus Tsai et Feng, Radix Notoginseng is saponin component, but Panax stipuleanatus Tsai et Feng
Main component is Triterpenoids sapogenins saponin(e in total saposins, hence it is evident that is different from dammarane type Fourth Ring three in arasaponin
Terpene saponin(e.The difference of saponin component type in Radix Notoginseng, Panax stipuleanatus Tsai et Feng medicinal material, will lead to the difference of its major function, can not
The two is used with.Currently, domestic and foreign scholars concentrate on its chemical component, implantation methods, Rapid identification to the research of Panax stipuleanatus Tsai et Feng
The research of method, nursing method, drying means, anti-inflammatory activity, anticancer activity and reducing blood lipid hypoglycemic activity etc., still
The antithrombotic disease of Panax stipuleanatus Tsai et Feng is not yet had been reported that.
Summary of the invention
In view of the problems of the existing technology the present invention, provides the preparation method and anticoagulation of a kind of Panax stipuleanatus Tsai et Feng total saposins
The application of drug, the present invention can effectively prepare high-purity Panax stipuleanatus Tsai et Feng total saposins, realize that Panax stipuleanatus Tsai et Feng treats thrombus disease in preparation
Application in medicine.
A kind of preparation method of Panax stipuleanatus Tsai et Feng total saposins, the specific steps are as follows:
Panax stipuleanatus Tsai et Feng powder is added in Extraction solvent, temperature be 60~100 DEG C under the conditions of Microwave Extraction 20~
40min obtains residue and extracting solution after filtering, then extracts 2~3 times under identical extraction conditions to residue, merges and extracts
Liquid, then be concentrated under reduced pressure and be dried to obtain medicinal extract, n-butanol medicinal extract is obtained using saturation extracting n-butyl alcohol medicinal extract, n-butanol medicinal extract uses
MCI pillar layer separation carries out ladder elution with ethanol/water dicyandiamide solution, collects volume fraction of ethanol in ethanol/water dicyandiamide solution
For 50%~90% ethanol eluate, ethanol eluate is concentrated, is drying to obtain Panax stipuleanatus Tsai et Feng total saposins.
The Panax stipuleanatus Tsai et Feng powder is the powder of Panax stipuleanatus Tsai et Feng main root and/or Panax stipuleanatus Tsai et Feng rhizome.
The solid-to-liquid ratio g:mL of the Panax stipuleanatus Tsai et Feng powder and Extraction solvent is 1:(5~30)；Extraction solvent be water, methanol,
Ethyl alcohol it is one or more.
Preferably, the solid-to-liquid ratio g:mL of the Panax stipuleanatus Tsai et Feng powder and Extraction solvent is 1:(15~20)；
Preferably, the Extraction solvent is ethanol-water solution or methanol-water solution, the volume of ethyl alcohol in ethanol-water solution
Content is 50%~70%；The volume content of methanol is 50%~70% in methanol-water solution.
The microwave power is 300~700W.
Preferably, the microwave power is 400~500W；
Preferably, Microwave Extraction temperature is 70~80 DEG C, and microwave extraction time is 20~30min；
Preferably, the ethanol eluate that volume fraction of ethanol is 60%~80% in ethanol/water dicyandiamide solution is collected.
Saponin(e mass content is 50%~90% in the Panax stipuleanatus Tsai et Feng total saposins；
Contain R in the Panax stipuleanatus Tsai et Feng total saposins1That is formula I and R2That is II compound represented of formula；
Panax stipuleanatus Tsai et Feng total saposins of the present invention can be used for preparing treatment thrombotic diseases and/or anticoagulant pharmaceutical composition
Object.
Described pharmaceutical composition includes one or more pharmaceutically acceptable auxiliary materials, and auxiliary material can be pharmaceutically acceptable
Carrier, diluent or excipient.
Described pharmaceutical composition is tablet, powder, granule, capsule, pill, suppository or aerosol.
The thrombotic diseases mainly include cerebrovascular thrombus, phlebothrombosis, thrombus of heart blood vessel etc..
The beneficial effects of the present invention are:
(1) Panax stipuleanatus Tsai et Feng total saposins prepared by the present invention are dissolved in water, purity is high, and preparation method is simple；
(2) high-purity Panax stipuleanatus Tsai et Feng total saposins prepared by the present invention, are natural drug, and toxic side effect is small, it can be achieved that screen side
Application of the Radix Notoginseng in preparation treatment thrombus disease drug.
Detailed description of the invention
Fig. 1 is the high-efficient liquid phase chromatogram of 1 Panax stipuleanatus Tsai et Feng total saposins of embodiment；Wherein 1- Panax stipuleanatus Tsai et Feng saponin(e R1, 2- screen
Side Panax Notoginseng saponin R2。
Specific embodiment
Invention is further described in detail With reference to embodiment, but protection scope of the present invention and unlimited
In the content.
A kind of embodiment 1: preparation method of Panax stipuleanatus Tsai et Feng total saposins, the specific steps are as follows:
By 200g Panax stipuleanatus Tsai et Feng powder (the main root and rhizome powder of Panax stipuleanatus Tsai et Feng), being added to 1L Extraction solvent, (alcohol-water is molten
Liquid) in, Microwave Extraction 20min under the conditions of temperature is 60 DEG C obtains residue and extracting solution, then in identical extraction item after filtering
Residue is extracted 3 times under part, combined extract, then is concentrated under reduced pressure and is dried to obtain medicinal extract i.e. Panax stipuleanatus Tsai et Feng extract
(79.4g) is obtained n-butanol medicinal extract (64.8g) using saturation extracting n-butyl alcohol medicinal extract, and n-butanol medicinal extract is using MCI column chromatography point
From (solvent system is ethyl alcohol by volume: water=30:70,40:60,50:50,60:40,70:30,80:20,90:10,100:0
Elution), with ethanol/water dicyandiamide solution carry out ladder elution, collect ethanol/water dicyandiamide solution in volume fraction of ethanol be 60%~
80% ethanol eluate, ethanol eluate is concentrated, is drying to obtain Panax stipuleanatus Tsai et Feng total saposins (51.5g)；Wherein Panax stipuleanatus Tsai et Feng
The solid-to-liquid ratio g:mL of powder and Extraction solvent is 1:5, and the volume content of ethyl alcohol is 50% in ethanol-water solution；Microwave power is
300W；
The high-efficient liquid phase chromatogram of Panax stipuleanatus Tsai et Feng total saposins is shown in Fig. 1；From fig. 1, it can be seen that with this condition, the total soap of Panax stipuleanatus Tsai et Feng
The chromatogram baseline of glycosides is steady, Panax stipuleanatus Tsai et Feng saponin(e R1And R2Ingredient separating degree is preferable；
Contain R in Panax stipuleanatus Tsai et Feng total saposins1That is formula I and R2That is II compound represented of formula；
Use in determined by ultraviolet spectrophotometry Panax stipuleanatus Tsai et Feng total saposins saponin content for 80.5%；Through high performance liquid chromatography
Method measurement, Panax stipuleanatus Tsai et Feng saponin(e R1And R2Content of the sum of the weight in Panax stipuleanatus Tsai et Feng total saposins is 54.8%.
A kind of embodiment 2: preparation method of Panax stipuleanatus Tsai et Feng total saposins, the specific steps are as follows:
By 200g Panax stipuleanatus Tsai et Feng powder (the main root and rhizome powder of Panax stipuleanatus Tsai et Feng), being added to 4L Extraction solvent, (alcohol-water is molten
Liquid) in, Microwave Extraction 25min under the conditions of temperature is 80 DEG C obtains residue and extracting solution, then in identical extraction item after filtering
Residue is extracted 3 times under part, combined extract, then is concentrated under reduced pressure and is dried to obtain medicinal extract i.e. Panax stipuleanatus Tsai et Feng extract
(98.4g) is obtained n-butanol medicinal extract (82.9g) using saturation extracting n-butyl alcohol medicinal extract, and n-butanol medicinal extract is using MCI column chromatography point
From (solvent system is ethyl alcohol by volume: water=30:70,40:60,50:50,60:40,70:30,80:20,90:10,100:0
Elution), with ethanol/water dicyandiamide solution carry out ladder elution, collect ethanol/water dicyandiamide solution in volume fraction of ethanol be 60%~
80% ethanol eluate, ethanol eluate is concentrated, is drying to obtain Panax stipuleanatus Tsai et Feng total saposins (63.6g)；Wherein Panax stipuleanatus Tsai et Feng
The solid-to-liquid ratio g:mL of powder and Extraction solvent is 1:20, and the volume content of ethyl alcohol is 60% in ethanol-water solution；Microwave power is
500W；
Contain R in Panax stipuleanatus Tsai et Feng total saposins1That is formula I and R2That is II compound represented of formula；
Use in determined by ultraviolet spectrophotometry Panax stipuleanatus Tsai et Feng total saposins saponin content for 82.6%；Through high performance liquid chromatography
Method measurement, Panax stipuleanatus Tsai et Feng saponin(e R1And R2Content of the sum of the weight in Panax stipuleanatus Tsai et Feng total saposins is 58.4%.
A kind of embodiment 3: preparation method of Panax stipuleanatus Tsai et Feng total saposins, the specific steps are as follows:
By 200g Panax stipuleanatus Tsai et Feng powder (the main root and rhizome powder of Panax stipuleanatus Tsai et Feng), being added to 6L Extraction solvent, (alcohol-water is molten
Liquid) in, Microwave Extraction 30min under the conditions of temperature is 100 DEG C obtains residue and extracting solution, then in identical extraction item after filtering
Residue is extracted 3 times under part, combined extract, then is concentrated under reduced pressure and is dried to obtain medicinal extract i.e. Panax stipuleanatus Tsai et Feng extract
(83.5g) is obtained n-butanol medicinal extract (66.4g) using saturation extracting n-butyl alcohol medicinal extract, and n-butanol medicinal extract is using MCI column chromatography point
From (solvent system is ethyl alcohol by volume: water=30:70,40:60,50:50,60:40,70:30,80:20,90:10,100:0
Elution), it is eluted with ethanol/water dicyandiamide solution, collecting volume fraction of ethanol in ethanol/water dicyandiamide solution is 60%~80%
Ethanol eluate, ethanol eluate is concentrated, is drying to obtain Panax stipuleanatus Tsai et Feng total saposins (54.2g)；Wherein Panax stipuleanatus Tsai et Feng powder
Solid-to-liquid ratio g:mL with Extraction solvent is 1:30, and the volume content of ethyl alcohol is 70% in ethanol-water solution；Microwave power is
300W；
Contain R in Panax stipuleanatus Tsai et Feng total saposins1That is formula I and R2That is II compound represented of formula；
Use in determined by ultraviolet spectrophotometry Panax stipuleanatus Tsai et Feng total saposins saponin content for 80.5%；Through high performance liquid chromatography
Method measurement, Panax stipuleanatus Tsai et Feng saponin(e R1And R2Content of the sum of the weight in Panax stipuleanatus Tsai et Feng total saposins is 54.1%.
Embodiment 4: Panax stipuleanatus Tsai et Feng saponin(e R in high effective liquid chromatography for measuring Panax stipuleanatus Tsai et Feng total saposins1And R2Content
Anticoagulant active component prepared by embodiment 1,2,3 uses high effective liquid chromatography for measuring Panax stipuleanatus Tsai et Feng saponin(e R1
And R2Content, it is specific as follows:
Instrument: Shimadzu high performance liquid chromatograph (including it is on-line degassing machine DGU-20A3R (C), binary pump LC-20AB, automatic
Sample injector SIL-20A, column oven CTO-20A, detector SPD-20A), chromatographic column: YMC-Triart C18 chromatographic column (5 μm,
250mm×4.6mm)；
Sample solution preparation:
Precision weighs Panax stipuleanatus Tsai et Feng total saposins powder 10mg, and 2mL70% methanol solution is added, through 0.45 μm of membrane filtration,
As test solution；
Chromatographic condition:
Chromatographic column: YMC-Triart C18 (250mm × 4.6mm, 5 μm) chromatographic column.Mobile phase A liquid is that 0.05% phosphoric acid is molten
Liquid；Mobile phase B liquid is acetonitrile.Mobile phase uses after ultrasonic degassing with 0.45 μm of membrane filtration.In 0~10min, by
10% Mobile phase B changes to 15% Mobile phase B, in 10~15min, changes to 20% Mobile phase B by 15% Mobile phase B,
In 15~28min, 25% Mobile phase B is changed to by 20% Mobile phase B, in 28~33min, is changed to by 25% Mobile phase B
30% Mobile phase B changes to 35% Mobile phase B by 30% Mobile phase B in 33~68min, in 68~81min, by
35% Mobile phase B changes to 75% Mobile phase B, in 81~96min, changes to 90% Mobile phase B by 75% Mobile phase B,
In 96~110min, 100% Mobile phase B is changed to by 90% Mobile phase B.Flow velocity is 1.0mLmin-1, column temperature is 30 DEG C, purple
Outer Detection wavelength is 203nm, and sample volume is 20.0 μ L；
It is linear to investigate:
Precision measures configured 2mg/mL Panax stipuleanatus Tsai et Feng saponin(e R1And R2Standard solution, be configured to 1.0 respectively, 0.8,
0.6, the serial standards solution of 0.4,0.2,0.1mg/mL carries out HPLC measurement by above-mentioned chromatographic condition, with Panax stipuleanatus Tsai et Feng soap
Glycosides R1And R2Concentration is abscissa, using peak area as ordinate, draws standard curve, equation of linear regression is Panax stipuleanatus Tsai et Feng saponin(e
R1: y=7 × 106x–12968(R2=0.9996), Panax stipuleanatus Tsai et Feng saponin(e R2: y=4 × 106x+23487(R2=0.9997)；
Precision test:
Take 0.4mg/mL Panax stipuleanatus Tsai et Feng saponin(e R1And R2Continuous 5 sample detections of standard solution, by corresponding peak area through marking
Its concentration of directrix curve equation calculation, as a result RSD value is respectively 0.42%, 0.51%；
Recovery test:
The Panax stipuleanatus Tsai et Feng saponin(e R of certain mass is added in the sample for taking known content1And R2, liquid is carried out by above-mentioned chromatographic condition
Mutually detect.Panax stipuleanatus Tsai et Feng saponin(e R1Average recovery rate be 104.23%, RSD value be 0.56%, Panax stipuleanatus Tsai et Feng saponin(e R2Be averaged
The rate of recovery is that 102.17%, RSD value is 0.39%.
Embodiment 5: the anticoagulant active of Panax stipuleanatus Tsai et Feng total saposins
Panax stipuleanatus Tsai et Feng total saposins are had studied to prothrombin time, activated partial thromboplastin time and fibrinogen
It influences；
Experimental method:
The preparation of mice plasma: taking Kunming mouse, plucks eyeball and takes blood, and the trisodium citrate of whole blood and 0.109mol/L are pressed
The ratio of 9:1 mixes, and 3500rpm/min is centrifuged 15min at 4 DEG C, collects supernatant, spare；
Sample preparation: taking Panax stipuleanatus Tsai et Feng extract and total saposins, is made into quality respectively with phosphate buffer (PBS) dissolution
Concentration is the solution of 5,10,20,30,50mg/mL.Taking heparin sodium is made into the solution that mass concentration is 1mg/mL；
The measurement of prothrombin time (PT), activated partial thromboplastin time (APTT) and fibrinogen (FIB):
The sample solution, heparin sodium aqua and appropriate PBS solution for taking various concentration, are added in appropriate blood plasma, according to reagent
Box illustrates step, and PT, APTT and FIB value are measured in coagulo meter.Blank group: PBS solution；Positive group: heparin sodium aqua (1
mg/mL)；
Experimental result:
It is screened using prothrombin time (PT), activated partial thromboplastin time (APTT) and fibrinogen (FIB)
The Anticoagulant Activities in vitro of test measurement Panax stipuleanatus Tsai et Feng extract and Panax stipuleanatus Tsai et Feng total saposins, the results are shown in Table 1.The total soap of Panax stipuleanatus Tsai et Feng
Glycosides is that Panax stipuleanatus Tsai et Feng extract is obtained by purifying.Compared to the blank group, when concentration is 30,50mg/mL, Panax stipuleanatus Tsai et Feng is total
Saponin(e group significantly extends PT, APTT, and Panax stipuleanatus Tsai et Feng extract group is without significant changes；
1 Panax stipuleanatus Tsai et Feng extract of table, total saposins are to prothrombin time (PT), activated partial thromboplastin time (APTT)
And fibrinogen (FIB) influence (N=3)
Compared with blank group,*P < 0.05,**P < 0.01,***p<0.001
As known from Table 1, compared to the blank group, Panax stipuleanatus Tsai et Feng extract and Panax stipuleanatus Tsai et Feng total saposins can drop to some extent
Low FIB content, activity are Panax stipuleanatus Tsai et Feng total saposins > Panax stipuleanatus Tsai et Feng extract；It is shown the result shows that Panax stipuleanatus Tsai et Feng total saposins have
The anticoagulant active of work.
Claims (9)
1. a kind of preparation method of Panax stipuleanatus Tsai et Feng total saposins, which is characterized in that specific step is as follows:
Panax stipuleanatus Tsai et Feng powder is added in Extraction solvent, 20 ~ 40 min of Microwave Extraction under the conditions of temperature is 60 ~ 100 DEG C, mistake
Residue and extracting solution are obtained after filter, then residue are extracted 2 ~ 3 times under identical extraction conditions, combined extract, then depressurize
Concentrate drying obtains medicinal extract, obtains n-butanol medicinal extract using saturation extracting n-butyl alcohol medicinal extract, n-butanol medicinal extract uses MCI column chromatography
Separation carries out ladder elution with ethanol/water dicyandiamide solution, and collecting volume fraction of ethanol in ethanol/water dicyandiamide solution is 50% ~ 90%
Ethanol eluate, ethanol eluate is concentrated, is drying to obtain Panax stipuleanatus Tsai et Feng total saposins.
2. the preparation method of Panax stipuleanatus Tsai et Feng total saposins according to claim 1, it is characterised in that: Panax stipuleanatus Tsai et Feng powder and extraction
The solid-to-liquid ratio g:mL of solvent is 1:(5 ~ 30)；Extraction solvent be water, methanol, ethyl alcohol it is one or more.
3. the preparation method of Panax stipuleanatus Tsai et Feng total saposins according to claim 2, it is characterised in that: Extraction solvent is alcohol-water
Solution or methanol-water solution, the volume content of ethyl alcohol is 50% ~ 70% in ethanol-water solution；The body of methanol in methanol-water solution
Product content is 50% ~ 70%.
4. the preparation method of Panax stipuleanatus Tsai et Feng total saposins according to claim 1, it is characterised in that: microwave power is 300 ~ 700
W。
5. the preparation method of Panax stipuleanatus Tsai et Feng total saposins according to claim 1, it is characterised in that: soap in Panax stipuleanatus Tsai et Feng total saposins
Glycosides mass content is 50% ~ 90%.
6. the preparation method of Panax stipuleanatus Tsai et Feng total saposins according to claim 1, it is characterised in that: contain in Panax stipuleanatus Tsai et Feng total saposins
There is II compound represented of formula I and formula:
。
7. Panax stipuleanatus Tsai et Feng total saposins prepared by the preparation method of any one of the claim 1 ~ 6 Panax stipuleanatus Tsai et Feng total saposins are being made
Application in standby treatment thrombotic diseases and/or anticoagulant pharmaceutical composition.
8. being applied described in claim 7, it is characterised in that: described pharmaceutical composition includes one or more pharmaceutically acceptable
Auxiliary material.
9. the application of claim 7 or 8, it is characterised in that: pharmaceutical composition is capsule, granule, tablet, powder or ball
Agent.
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| CN111494447A (en) * | 2020-06-23 | 2020-08-07 | 云南中医药大学 | Drug-resistant fungus resistant pharmaceutical composition and preparation thereof |
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| CN111494447A (en) * | 2020-06-23 | 2020-08-07 | 云南中医药大学 | Drug-resistant fungus resistant pharmaceutical composition and preparation thereof |
| CN111494447B (en) * | 2020-06-23 | 2022-05-27 | 云南中医药大学 | Drug-resistant fungus resistant pharmaceutical composition and preparation thereof |
| CN112410321A (en) * | 2020-11-26 | 2021-02-26 | 昆明理工大学 | Beta-glucosidase Ttbgl3 and application thereof |
| CN112410321B (en) * | 2020-11-26 | 2022-01-28 | 昆明理工大学 | Beta-glucosidase Ttbgl3 and application thereof |
| CN112656829A (en) * | 2020-12-31 | 2021-04-16 | 西南林业大学 | Method for simultaneously extracting notoginsenoside, notoginsenones and notoginsen polysaccharide and application thereof |
| CN112656829B (en) * | 2020-12-31 | 2022-04-01 | 西南林业大学 | Method for simultaneously extracting notoginsenoside, notoginsenones and notoginsen polysaccharide and application thereof |
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