CN105147804B - A kind of radix sanguisorbae total saponin extract and its preparation method and application - Google Patents
A kind of radix sanguisorbae total saponin extract and its preparation method and application Download PDFInfo
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Abstract
The present invention provides a kind of radix sanguisorbae total saponin extracts and its preparation method and application.In radix sanguisorbae total saponin extract prepared by the present invention, total saponin content reaches 98.50%.Compared with existing method, radix sanguisorbae total saponin content prepared by the present invention is dramatically increased, and the activity of increasing leukocyte is also obviously improved.Present invention process is easy to operate, high-efficient, and production cost is low, is conducive to the industrialized production of radix sanguisorbae total saponin.The activity of radix sanguisorbae total saponin extract increasing leukocyte prepared by the present invention is strong, provides a kind of new drug for the clinical treatment of leukopenia.
Description
Technical field
The present invention relates to a kind of radix sanguisorbae total saponin extracts and its preparation method and application, belong to drug field.
Background technique
Often there is myelosuppression during tumour Radiotherapy chemotherapy, so as to cause leukopenia.Leukocyte count subtracts
It is few, human immunological competence can be weakened, serious infection and bleeding often occurs in clinic.Therefore, it treats white caused by tumour Radiotherapy chemotherapy
Cytopenia needs to set about from protection hemopoietic function of bone marrow.
Clinical history of the garden burnet as prescribed preparation for existing more than 20 years of the treatment of bone marrow suppression, has taking dose
Small, significant in efficacy, safe and non-toxic, from a wealth of sources, the advantages that preparation process is simple, cheap, therefore be expected to be developed to face
Bed prevents and treats the irreplaceable natural drug of bone marrow suppression.
At present it has proven convenient that it is mainly two substance of saponin(e and tannin that garden burnet, which promotees the material base that haemocyte generates,.Tannin
Method for separating and preparing and technology relative maturity, and do not have a kind of preferably to isolate and purify high level at present in production
The method of radix sanguisorbae total saponin.
Application number: 200910082470.6, denomination of invention: the preparation method of radix sanguisorbae total saponin and sanguisorbin I, this method
It needs to be isolated and purified by macroreticular resin, higher cost, low efficiency is unfavorable for industrial production.Application number:
201210129134.4 denominations of invention: the isolation and purification method of radix sanguisorbae total saponin, this method need to take off garden burnet extracting solution
Rouge, precipitating, extracts at extraction again, and during the preparation process, extracting solution is repeatedly shifted, and big, complex steps are lost in radix sanguisorbae total saponin,
Also it is unfavorable for industrial production.
Therefore it provides a kind of technological operation radix sanguisorbae total saponin content that is easy, at low cost, high-efficient, being prepared it is high and
The preparation method being suitable for industrialized large-scaled production has particularly important meaning to the development and application of garden burnet resource.
Summary of the invention
The technical solution of the present invention is to provide a kind of radix sanguisorbae total saponin extracts and its preparation method and application.
The present invention provides a kind of radix sanguisorbae total saponin extract, sanguisorbin I content in the extract are as follows: 50%-97%,
Sanguisorbin II content are as follows: 2%-10%.
Preferably, sanguisorbin I content in the extract are as follows: 92.34%, sanguisorbin II content are as follows: 6.16%.
Wherein, total saposins weight percentage is 60-99% in the extract.
Preferably, total saposins weight percentage is 98.50% in the extract.
Wherein, the extract, is prepared as follows:
A. garden burnet medicinal material is taken, is extracted added with solvent, extracting solution is spare；
B. extracting solution made from step a is taken, pH to 12~14 is adjusted, is stood, centrifugation filters to obtain supernatant, supernatant decompression
Concentration, it is spare；
C. supernatant tune pH to 11~12 made from step b is taken, is stood, precipitating is collected in centrifugation；Supernatant continues to adjust pH extremely
12~13, it stands, precipitating is collected in centrifugation；Merging precipitates twice；
D. step c precipitating obtained is taken to extract added with solvent, filtrate is collected in filtering, concentrating filter liquor, it is dry to get
Radix sanguisorbae total saponin.
The present invention provides a kind of methods for preparing radix sanguisorbae total saponin extract, it includes the following steps:
A. garden burnet medicinal material is taken, is extracted added with solvent, extracting solution is spare；
B. extracting solution made from step a is taken, pH to 12~14 is adjusted, is stood, centrifugation filters to obtain supernatant, supernatant decompression
Concentration, it is spare；
C. supernatant tune pH to 11~12 made from step b is taken, is stood, precipitating is collected in centrifugation；Supernatant continues to adjust pH extremely
12~13, it stands, precipitating is collected in centrifugation；Merging precipitates twice；
D. step c precipitating obtained is taken to extract added with solvent, filtrate is collected in filtering, concentrating filter liquor, it is dry to get
Radix sanguisorbae total saponin.
Wherein, in step a, add organic solvent refluxing extraction 2 times, every time 1.5 hours of 8 times of garden burnet medicinal material amounts；Described
Organic solvent is ethyl alcohol or methanol, and concentration of alcohol is 30%~95%v/v.
Preferably, the organic solvent is ethyl alcohol, concentration of alcohol 90%.
Wherein, in step b, extracting solution adds 10% aqueous slkali tune pH to 12~14, stands overnight, and centrifugation filters
Clear liquid, supernatant are concentrated under reduced pressure into proper volume, and adding appropriate amount of water to concentration of alcohol is 20%v/v, spare；The aqueous slkali is
NaOH, KOH or Ca (OH)2Solution.
Preferably, the aqueous slkali is NaOH solution.
In step c, time of repose 12h, collection is deposited in 70 DEG C and is dried under reduced pressure.
In step d, step c precipitating obtained is taken, with organic solvent reflux 45min, filtering is collected filtrate, is recovered under reduced pressure
Organic solvent is volatilized to there is solid content precipitation, and gained solid content is dried under reduced pressure 12 hours to get radix sanguisorbae total saponin；Wherein, described
Organic solvent be dehydrated alcohol or anhydrous methanol.
Preferably, the organic solvent is dehydrated alcohol.
The present invention provides use of the radix sanguisorbae total saponin extract in the drug that preparation has function of increasing leukocyte
On the way.
The present invention also provides a kind of pharmaceutical compositions of increasing leukocyte, it is by the radix sanguisorbae total saponin extract
For active constituent, pharmaceutically acceptable auxiliary material is added or complementary ingredient is prepared into pharmaceutically common preparation.
Wherein, the preparation is oral preparation.
The present invention substantially increases containing for radix sanguisorbae total saponin by the improvement to radix sanguisorbae total saponin extract preparation method
Amount, makes its content reach 98.50%.Compared with existing method, radix sanguisorbae total saponin is contained in terms of sanguisorbin I and sanguisorbin II
Amount dramatically increases, and the activity of increasing leukocyte is also obviously improved.Present invention process is easy to operate, high-efficient, production
It is at low cost, be conducive to the industrialized production of radix sanguisorbae total saponin.Radix sanguisorbae total saponin prepared by the present invention is leukopenia
Clinical treatment provides a kind of new drug.
Below by way of specific embodiment, the present invention is described in further detail, but is not intended to limit the present invention, ability
The various changes and replacement that field technique personnel make according to the present invention, as long as it does not depart from the spirit of the invention, should belong to this hair
Bright scope of the appended claims.
Specific embodiment
The preparation method of 1 radix sanguisorbae total saponin extract of embodiment
Garden burnet medicine materical crude slice 1kg is taken, after appropriate crushing plus 8 times of 90% alcohol refluxs of amount extract 2 times, it 1.5 hours every time, filters,
Merge extracting solution twice to add 10% NaOH solution tune pH to 12~14 after standing is let cool, stand overnight, centrifugation goes to precipitate, and filters
Liquid filters to obtain supernatant.Supernatant is concentrated under reduced pressure to proper volume, adds appropriate amount of water to concentration of alcohol 20% or so, with 10%
NaOH solution tune pH to 11~12, stand 12h, precipitating is collected by centrifugation；Supernatant continue with 10% NaOH solution tune pH extremely
12~13,12h is stood, precipitating is collected by centrifugation；Merge two sides precipitating, be decompressed in 70 DEG C it is dry, with dehydrated alcohol flow back 45min,
Filtering, collect filtrate, ethyl alcohol is recovered under reduced pressure to there is solid content precipitation, volatilizes, gained solid content be dried under reduced pressure 12 hours to get.
The preparation method of 2 radix sanguisorbae total saponin extract of embodiment
Garden burnet medicine materical crude slice 1kg is taken, after appropriate crushing plus 8 times of 90% methanol eddies of amount extract 2 times, it 1.5 hours every time, filters,
Merge extracting solution twice to add 10% NaOH solution tune pH to 12~14 after standing is let cool, stand overnight, centrifugation goes to precipitate, and filters
Liquid filters to obtain supernatant.Supernatant is concentrated under reduced pressure to proper volume, adds appropriate amount of water to methanol concentration 20% or so, with 10%
NaOH solution tune pH to 11~12, stand 12h, precipitating is collected by centrifugation；Supernatant continue with 10% NaOH solution tune pH extremely
12~13,12h is stood, precipitating is collected by centrifugation；Merge two sides precipitating, be decompressed in 70 DEG C it is dry, with anhydrous methanol flow back 45min,
Filtering, collect filtrate, methanol is recovered under reduced pressure to there is solid content precipitation, volatilizes, gained solid content be dried under reduced pressure 12 hours to get.
The preparation method of 3 radix sanguisorbae total saponin extract of embodiment
Garden burnet medicine materical crude slice 1kg is taken, after appropriate crushing plus 8 times of 90% alcohol refluxs of amount extract 2 times, it 1.5 hours every time, filters,
Merge extracting solution twice to add 10% KOH solution tune pH to 12~14 after standing is let cool, stand overnight, centrifugation goes to precipitate, and filters
Liquid filters to obtain supernatant.Supernatant is concentrated under reduced pressure to proper volume, adds appropriate amount of water to concentration of alcohol 20% or so, with 10%
KOH solution tune pH to 11~12, stand 12h, precipitating is collected by centrifugation；Supernatant continues the KOH solution tune pH to 12 with 10%
~13,12h is stood, precipitating is collected by centrifugation；Merge two sides precipitating, be decompressed in 70 DEG C it is dry, with dehydrated alcohol flow back 45min, mistake
Filter, collect filtrate, ethyl alcohol is recovered under reduced pressure to there is solid content precipitation, volatilizes, gained solid content be dried under reduced pressure 12 hours to get.
The preparation method of 4 radix sanguisorbae total saponin extract of embodiment
Garden burnet medicine materical crude slice 1kg is taken, after appropriate crushing plus 8 times of 90% alcohol refluxs of amount extract 2 times, it 1.5 hours every time, filters,
Merge extracting solution twice and adds 10% Ca (OH) after standing is let cool2Solution tune pH to 12~14, stands overnight, and centrifugation goes to sink
It forms sediment, filtrate filters to obtain supernatant.Supernatant be concentrated under reduced pressure to proper volume, add appropriate amount of water to concentration of alcohol 20% or so,
With 10% Ca (OH)2Solution tune pH to 11~12 stands 12h, precipitating is collected by centrifugation；Supernatant continues the Ca (OH) with 10%2
Solution tune pH to 12~13 stands 12h, precipitating is collected by centrifugation；Merge two sides precipitating, is decompressed in 70 DEG C dry, uses dehydrated alcohol
Flow back 45min, and filtering collects filtrate, ethyl alcohol is recovered under reduced pressure to there is solid content precipitation, volatilizes, gained solid content is dried under reduced pressure 12
Hour to get.
The preparation method of 5 radix sanguisorbae total saponin extract of embodiment
Garden burnet medicine materical crude slice 1kg is taken, after appropriate crushing plus 8 times of 30% alcohol refluxs of amount extract 2 times, it 1.5 hours every time, filters,
Merge extracting solution twice to add 10% NaOH solution tune pH to 12~14 after standing is let cool, stand overnight, centrifugation goes to precipitate, and filters
Liquid filters to obtain supernatant.Appropriate amount of water is added to be stood to concentration of alcohol 20% or so with 10% NaOH solution tune pH to 11~12
Precipitating is collected by centrifugation in 12h；Supernatant continues the NaOH solution tune pH to 12~13 with 10%, stands 12h, precipitating is collected by centrifugation；
Merge two sides precipitating, be decompressed in 70 DEG C it is dry, with dehydrated alcohol flow back 45min, filtering, collect filtrate, ethyl alcohol is recovered under reduced pressure extremely
Have solid content precipitation, volatilize, gained solid content be dried under reduced pressure 12 hours to get.
The preparation method of 6 radix sanguisorbae total saponin extract of embodiment
Garden burnet medicine materical crude slice 1kg is taken, after appropriate crushing plus 8 times of 40% alcohol refluxs of amount extract 2 times, it 1.5 hours every time, filters,
Merge extracting solution twice to add 10% NaOH solution tune pH to 12~14 after standing is let cool, stand overnight, centrifugation goes to precipitate, and filters
Liquid filters to obtain supernatant.Appropriate amount of water is added to be stood to concentration of alcohol 20% or so with 10% NaOH solution tune pH to 11~12
Precipitating is collected by centrifugation in 12h；Supernatant continues the NaOH solution tune pH to 12~13 with 10%, stands 12h, precipitating is collected by centrifugation；
Merge two sides precipitating, be decompressed in 70 DEG C it is dry, with dehydrated alcohol flow back 45min, filtering, collect filtrate, ethyl alcohol is recovered under reduced pressure extremely
Have solid content precipitation, volatilize, gained solid content be dried under reduced pressure 12 hours to get.
The preparation method of 7 radix sanguisorbae total saponin extract of embodiment
Garden burnet medicine materical crude slice 1kg is taken, after appropriate crushing plus 8 times of 50% alcohol refluxs of amount extract 2 times, it 1.5 hours every time, filters,
Merge extracting solution twice to add 10% NaOH solution tune pH to 12~14 after standing is let cool, stand overnight, centrifugation goes to precipitate, and filters
Liquid filters to obtain supernatant.Supernatant is concentrated under reduced pressure to proper volume, adds appropriate amount of water to concentration of alcohol 20% or so, with 10%
NaOH solution tune pH to 11~12, stand 12h, precipitating is collected by centrifugation；Supernatant continue with 10% NaOH solution tune pH extremely
12~13,12h is stood, precipitating is collected by centrifugation；Merge two sides precipitating, be decompressed in 70 DEG C it is dry, with dehydrated alcohol flow back 45min,
Filtering, collect filtrate, ethyl alcohol is recovered under reduced pressure to there is solid content precipitation, volatilizes, gained solid content be dried under reduced pressure 12 hours to get.
The preparation method of 8 radix sanguisorbae total saponin extract of embodiment
Garden burnet medicine materical crude slice 1kg is taken, after appropriate crushing plus 8 times of 60% alcohol refluxs of amount extract 2 times, it 1.5 hours every time, filters,
Merge extracting solution twice to add 10% NaOH solution tune pH to 12~14 after standing is let cool, stand overnight, centrifugation goes to precipitate, and filters
Liquid filters to obtain supernatant.Supernatant is concentrated under reduced pressure to proper volume, adds appropriate amount of water to concentration of alcohol 20% or so, with 10%
NaOH solution tune pH to 11~12, stand 12h, precipitating is collected by centrifugation；Supernatant continue with 10% NaOH solution tune pH extremely
12~13,12h is stood, precipitating is collected by centrifugation；Merge two sides precipitating, be decompressed in 70 DEG C it is dry, with dehydrated alcohol flow back 45min,
Filtering, collect filtrate, ethyl alcohol is recovered under reduced pressure to there is solid content precipitation, volatilizes, gained solid content be dried under reduced pressure 12 hours to get.
The preparation method of 9 radix sanguisorbae total saponin extract of embodiment
Garden burnet medicine materical crude slice 1kg is taken, after appropriate crushing plus 8 times of 70% alcohol refluxs of amount extract 2 times, it 1.5 hours every time, filters,
Merge extracting solution twice to add 10% NaOH solution tune pH to 12~14 after standing is let cool, stand overnight, centrifugation goes to precipitate, and filters
Liquid filters to obtain supernatant.Supernatant is concentrated under reduced pressure to proper volume, adds appropriate amount of water to concentration of alcohol 20% or so, with 10%
NaOH solution tune pH to 11~12, stand 12h, precipitating is collected by centrifugation；Supernatant continue with 10% NaOH solution tune pH extremely
12~13,12h is stood, precipitating is collected by centrifugation；Merge two sides precipitating, be decompressed in 70 DEG C it is dry, with dehydrated alcohol flow back 45min,
Filtering, collect filtrate, ethyl alcohol is recovered under reduced pressure to there is solid content precipitation, volatilizes, gained solid content be dried under reduced pressure 12 hours to get.
The preparation method of 10 radix sanguisorbae total saponin extract of embodiment
Garden burnet medicine materical crude slice 1kg is taken, after appropriate crushing plus 8 times of 80% alcohol refluxs of amount extract 2 times, it 1.5 hours every time, filters,
Merge extracting solution twice to add 10% NaOH solution tune pH to 12~14 after standing is let cool, stand overnight, centrifugation goes to precipitate, and filters
Liquid filters to obtain supernatant.Supernatant is concentrated under reduced pressure to proper volume, adds appropriate amount of water to concentration of alcohol 20% or so, with 10%
NaOH solution tune pH to 11~12, stand 12h, precipitating is collected by centrifugation；Supernatant continue with 10% NaOH solution tune pH extremely
12~13,12h is stood, precipitating is collected by centrifugation；Merge two sides precipitating, be decompressed in 70 DEG C it is dry, with dehydrated alcohol flow back 45min,
Filtering, collect filtrate, ethyl alcohol is recovered under reduced pressure to there is solid content precipitation, volatilizes, gained solid content be dried under reduced pressure 12 hours to get.
The preparation method of 11 radix sanguisorbae total saponin extract of embodiment
Garden burnet medicine materical crude slice 1kg is taken, after appropriate crushing plus 8 times of 95% alcohol refluxs of amount extract 2 times, it 1.5 hours every time, filters,
Merge extracting solution twice to add 10% NaOH solution tune pH to 12~14 after standing is let cool, stand overnight, centrifugation goes to precipitate, and filters
Liquid filters to obtain supernatant.Supernatant is concentrated under reduced pressure to proper volume, adds appropriate amount of water to concentration of alcohol 20% or so, with 10%
NaOH solution tune pH to 11~12, stand 12h, precipitating is collected by centrifugation；Supernatant continue with 10% NaOH solution tune pH extremely
12~13,12h is stood, precipitating is collected by centrifugation；Merge two sides precipitating, be decompressed in 70 DEG C it is dry, with dehydrated alcohol flow back 45min,
Filtering, collect filtrate, ethyl alcohol is recovered under reduced pressure to there is solid content precipitation, volatilizes, gained solid content be dried under reduced pressure 12 hours to get.
Beneficial effects of the present invention are proved below by way of specific experiment.
The comparative experiments of the preparation method and patented method of the radix sanguisorbae total saponin extract of the present invention of experimental example 1
1. experimental method:
The preparation of 1.1 laboratory samples
Sample 1 is prepared according to embodiment 1.
According to following patents, (application number: 200910082470.6) method a prepares sample 2.
According to following patents, (application number: 201210129134.4) method b prepares sample 3.
Patented method a: taking garden burnet medicine materical crude slice 1kg, and 8 times of 30% ethyl alcohol heating and refluxing extraction 1h of amount are added, and filter residue uses 6 times of amounts again
30% ethyl alcohol heating and refluxing extraction 1h, merging filtrate are added 20 times of amount water dilutions, are added with stirring sodium hydroxide solution (3.5kg
Sodium hydroxide is dissolved in 24L water), adjusting pH value is 14.2, stands, is centrifuged to obtain filtrate, passes through D101 type macroporous absorbent resin, flow velocity
For 1.5 column volumes/h, again with 4 30% ethanol elutions of column volume after upper complete medical fluid, be then washed with water take off it is colourless to efflux,
Finally with 70% ethanol elution of 3 times of bed volumes, 70% ethyl alcohol efflux is collected, measured quantity of material precipitation has been concentrated in vacuo to
Afterwards, stop concentration, stand 2 hours or more, decompression filters, and with 40% ethanol washing, drains, and takes out filter cake, vacuum drying, powder
It is broken to get.
Patented method b: taking garden burnet medicine materical crude slice 1kg, extracts 3 times, each 1.5h, 70 DEG C decompressions with 8 times of 70% alcohol refluxs of amount
Concentrated medicament is transferred to separatory funnel, the ether defatting being first saturated with water 2 times, 70 DEG C of mother liquor subtract to appropriate (no ethyl alcohol taste)
It pushes back and receives mixed ether, then plus water is dissolved to, then extracting n-butyl alcohol 2 times be saturated with water, 80 DEG C of extract liquor reduced pressures,
Add certain density ethyl alcohol to dissolve solid content, be transferred in beaker, add appropriate amount of water, adjust pH to 10-14, place, be centrifuged, takes out
Precipitating, 70 DEG C of vacuum drying 2h are transferred in triangular flask, add dehydrated alcohol ultrasonic extraction 30 minutes, filter, 70 DEG C of filtrate
Ethyl alcohol is recovered under reduced pressure to just having solid content precipitation, is transferred in evaporating dish, 80 DEG C of water-baths volatilize ethyl alcohol, 70 DEG C of vacuum drying 12h,
It takes out, grinds to get radix sanguisorbae total saponin.
The measurement of 1.2 sanguisorbin I contents:
Using the method for high performance liquid chromatography, condition is Sapphire C18Column (5 μm, 150 × 4.6mm) chromatographic column point
From being filler with octadecylsilane chemically bonded silica；With methanol-water (64:36) for mobile phase；Flow velocity 1.0mL/min, ELSD
(evaporative light scattering detector) detection, 85 DEG C of drift tube temperature, gas flow rate 2.8L/min.Theoretical cam curve presses sanguisorbin I
Peak calculates, and should be not less than 4000.
The preparation of reference substance solution in experiment: it is appropriate that precision weighs sanguisorbin I reference substance, is dissolved respectively with 50% ethyl alcohol
Solution of every 1mL containing 0.0565mg and 0.226mg is made, it is spare.
The preparation of test solution: taking medicinal extract (dry) 10g or 30, preparation, and accurately weighed weight is finely ground, takes in right amount (about
It is equivalent to 10 weight) totally 2 parts, it is accurately weighed, it sets in 150mL stuffed conical flask, 50% ethyl alcohol 20mL is added in precision, shakes
Even, weighing, ultrasound 10 minutes puts to room temperature, the weight of less loss is supplied with 50% ethyl alcohol, is shaken up, and is centrifuged, takes supernatant, 0.22 μ
The filtration of m filter membrane, takes subsequent filtrate as test solution.Then reference substance solution and each 20 μ l of test solution are taken respectively, by upper
Chromatographic condition measurement is stated, and calculates sanguisorbin I content in sample by external standard two-point method.
The measurement of 1.3 sanguisorbin, II content:
Using the method for high performance liquid chromatography, condition is Yi Lite Hypersil C18Chromatographic column (4.6mm × 200mm, 5
μm)；Volume flow 1.2mL/min；Mobile phase A is methanol-acetonitrile (1: 1), and Mobile phase B is 0.5% phosphoric acid solution, gradient elution
(0~10min, 45.0%A；10~27min, 45.0%-60.0%A；27~40min, 60.0%A；40~45min,
60.0%-45.0%A)；Detection wavelength (0~27min, λ1=203nm；27~35min, λ2=289nm；35~45min, λ3=
306nm)；10 μ L of sample volume.Theoretical cam curve must not be lower than 3000, and separating degree is all larger than 2.
The preparation of reference substance solution in experiment: it is appropriate that precision weighs-II reference substance of sanguisorbin, is respectively placed in 5 20mL
In measuring bottle, add 60% methanol to dissolve and be diluted to scale, then respectively it is accurate draw above-mentioned solution 10,5,2,10,5mL be placed in 50mL
In measuring bottle, adds 60% methanol dilution to scale, shake up-II reference substance solution of sanguisorbin to get 0.0374mg/mL.
The preparation of test solution: taking medicinal extract (dry) in right amount, finely ground, mixes, weighs about 1.0g, accurately weighed, accurate
It is added 60% methanol 50mL, close plug, weighed quality, ultrasonic wave (power 160W, frequency 50kHz) is ultrasonically treated 30min, lets cool,
Weighed quality again is supplied the quality of less loss with 60% methanol, is shaken up, and filtering takes subsequent filtrate as test solution.Then accurate
Reference substance solution and test solution are drawn, is measured by above-mentioned chromatographic condition, and calculates garden burnet soap in sample by external standard two-point method
- II content of glycosides.
2. experimental result:
The comparision contents of 1 radix sanguisorbae total saponin of table
3. experiment conclusion:
Experimental data is shown, compared with two kinds of patented methods, sanguisorbin I, sanguisorbin II that the present invention is prepared
It is higher with radix sanguisorbae total saponin and yield, sufficiently prove that the method for the present invention substantially increases radix sanguisorbae total saponin, sanguisorbin I and ground
The content of elm saponin I I, hence it is evident that better than the method for patent disclosure.
The pharmacological evaluation of the radix sanguisorbae total saponin extract of the present invention of experimental example 2 compares
1. experimental material, instrument
1.1 test medicines: sample 1, sample 2, the sample 3 prepared in experimental example 1.
1.2 tool drugs: cyclophosphamide.
1.3 experimental animals: KM mouse: 18.5~22.5g of weight.
1.4 laboratory apparatus: full-automatic blood cell analysis machine；BS-600L electronic balance: specification: 600g/0.1g, Shanghai friend's sound
Weighing apparatus Co., Ltd.
1.5 statistical methods: for statistical analysis with 17.0 software of SPSS.Data are with mean ± standard deviationIt indicates,
One-way analysis of variance is used between group, LSD inspection is carried out between the neat person's group of variance, and heterogeneity of variance person carries out Tamhane ' s T2 inspection
It tests.
2. experimental method
The grouping of 2.1 experimental animals and model preparation:
All Animal adaptabilities are randomly divided into after feeding 1 week by weight: normal group, model group, sample 1 group: 15mgkg-
1 dosage stomach-filling sample, 1 aqueous solution；2 groups: 15mgkg-1 stomach-filling sample of sample, 2 aqueous solution；3 groups: 15mgkg-1 dosage of sample
3 aqueous solution of stomach-filling sample；Diyushengbai Tablet group: 200mgkg-1 dosage stomach-filling Diyushengbai Tablet aqueous solution, every group 10.It is real
It tests the 1st day, in addition to blank group, remaining each group mouse presses 50mgkg-1 dosage intraperitoneal injection of cyclophosphamide normal saline solution,
For three days on end, isometric physiological saline is injected intraperitoneally in naive mice.
2.2 experimental groups and administration:
Each experimental group started since experiment the 1st day according to dosage, administration mode give relative medicine, normal group and model group mouse
The isometric pure water of stomach-filling, continuous 7 days.
2.3 collections of specimens:
It tests the 8th day, each experimental mice eye socket takes blood, is collected with the 0.5mlEP pipe equipped with EDTA anti-coagulants to be measured.
2.4 Testing index and method:
Peripheral hemogram detection: each experimental mice peripheral white blood cells (WBC) are carried out using full-automatic blood counting instrument
It counts.
3. experimental result
Each experimental mice peripheral white blood cell amount of table 2
Note: compared with model group, P < 0.01 * P < 0.05, * *；Compared with Diyushengbai Tablet group,▲P < 0.05,▲▲P<0.01。
4. experiment conclusion
Table 2 shows that 1 group of mouse peripheral blood leucocyte of sample has significant raising (P < 0.05) compared with model group；Even than sun
Property group Diyushengbai Tablet group it is also high, show that the activity of radix sanguisorbae total saponin increasing leukocyte prepared by the present invention is best.
To sum up, compared with existing patented method, preparation method of the present invention significantly improves radix sanguisorbae total saponin, sanguisorbin I
With the content of sanguisorbin II, and drug effect be also significantly better than existing method preparation total saposins.Preparation method technique behaviour of the present invention
Work is easy, at low cost, high-efficient, is suitable for industrialized large-scaled production.Radix sanguisorbae total saponin extract increasing leukocyte prepared by the present invention
Activity it is strong, provide the new drug of one kind for the clinical treatment of leukopenia.
Claims (6)
1. a kind of radix sanguisorbae total saponin extract, it is characterised in that: the weight percentage of sanguisorbin I in the extract are as follows:
92.34%, the weight percentage of sanguisorbin II are as follows: 6.16%.
2. extract according to claim 1, it is characterised in that: total saposins weight percentage is in the extract
98.50%.
3. extract according to claim 1 or 2, it is characterised in that: it is to prepare by the following method:
A. garden burnet medicinal material is taken, is extracted added with solvent, extracting solution is spare；
B. extracting solution made from step a is taken, pH to 12~14 is adjusted, is stood, centrifugation filters to obtain supernatant, and supernatant is concentrated under reduced pressure,
It is spare；
C. reserve liquid tune pH to 11~12 made from step b is taken, is stood, precipitating is collected in centrifugation；Supernatant continue adjust pH to 12~
13, it stands, precipitating is collected in centrifugation；Merging precipitates twice；
D. precipitating made from step c is taken to extract added with solvent, filtrate is collected in filtering, and concentrating filter liquor, drying are to get garden burnet
Total saposins；
Wherein,
In step a, the organic solvent is ethyl alcohol, concentration of alcohol 90%v/v；
In step d, the organic solvent is dehydrated alcohol.
4. the method for preparing extract described in claim 1-3 any one, it includes the following steps:
A. garden burnet medicinal material is taken, is extracted added with solvent, extracting solution is spare；
B. extracting solution made from step a is taken, 10% aqueous slkali tune pH to 12~14 is added, is stood, centrifugation filters to obtain supernatant,
Supernatant is concentrated under reduced pressure, spare；
C. it takes reserve liquid made from step b to add 10% aqueous slkali tune pH to 11~12, stands, precipitating is collected in centrifugation；Supernatant
Continue plus 10% aqueous slkali tune pH to 12~13, stand, centrifugation, collect precipitating；Merging precipitates twice；
D. precipitating made from step c is taken to extract added with solvent, filtrate is collected in filtering, and concentrating filter liquor, drying are to get garden burnet
Total saposins；
Wherein,
In step a, the organic solvent is ethyl alcohol, concentration of alcohol 90%v/v；
In step d, the organic solvent is dehydrated alcohol.
5. according to the method described in claim 4, it is characterized by:
In step a, add organic solvent refluxing extraction 2 times, every time 1.5 hours of 8 times of garden burnet medicinal material amounts；
In step b, extracting solution adds 10% aqueous slkali tune pH to 12~14, stands overnight, and centrifugation filters to obtain supernatant, supernatant
Liquid is concentrated under reduced pressure into proper volume, and adding appropriate amount of water to concentration of alcohol is 20%v/v, spare；Wherein, the aqueous slkali is
NaOH, KOH or Ca (OH)2Solution；
In step c, time of repose 12h, collection is deposited in 70 DEG C and is dried under reduced pressure；In step d, step c precipitating obtained is taken,
With organic solvent reflux 45min, filtering collects filtrate, organic solvent is recovered under reduced pressure to there is solid content precipitation, volatilizes, gained is solid
Shape object is dried under reduced pressure 12 hours to get radix sanguisorbae total saponin.
6. according to the method described in claim 5, it is characterized by: the aqueous slkali is that NaOH is molten in step b and step c
Liquid.
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| CN105920137A (en) * | 2016-06-02 | 2016-09-07 | 中国药科大学 | Application of sanguisorba total saponin to treatment of inflammatory bowel diseases |
| CN107661287B (en) * | 2017-11-28 | 2021-01-26 | 遵义医学院 | Preparation of sanguisorbin self-emulsifying drug delivery system |
| CN109975437A (en) * | 2017-12-27 | 2019-07-05 | 四川科瑞德制药股份有限公司 | The analysis method of sanguisorbin I in garden burnet or product containing garden burnet |
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| CN101810705A (en) * | 2009-05-26 | 2010-08-25 | 成都地奥集团天府药业股份有限公司 | Content determination method of sanguisorbin I |
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| CN101862385A (en) * | 2009-04-20 | 2010-10-20 | 成都地奥制药集团有限公司 | Sanguisorba saponins and preparation method of sanguisorbin I |
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