CN106831936B - The method that tanshinone IIA and dihydrotanshinone I are prepared using middle high-pressure preparation liquid phase method - Google Patents
The method that tanshinone IIA and dihydrotanshinone I are prepared using middle high-pressure preparation liquid phase method Download PDFInfo
- Publication number
- CN106831936B CN106831936B CN201710026412.6A CN201710026412A CN106831936B CN 106831936 B CN106831936 B CN 106831936B CN 201710026412 A CN201710026412 A CN 201710026412A CN 106831936 B CN106831936 B CN 106831936B
- Authority
- CN
- China
- Prior art keywords
- liquid phase
- dihydrotanshinone
- tanshinone iia
- phase
- extract
- 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.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J73/00—Steroids in which the cyclopenta[a]hydrophenanthrene skeleton has been modified by substitution of one or two carbon atoms by hetero atoms
- C07J73/001—Steroids in which the cyclopenta[a]hydrophenanthrene skeleton has been modified by substitution of one or two carbon atoms by hetero atoms by one hetero atom
- C07J73/003—Steroids in which the cyclopenta[a]hydrophenanthrene skeleton has been modified by substitution of one or two carbon atoms by hetero atoms by one hetero atom by oxygen as hetero atom
Abstract
The invention discloses a kind of methods for preparing tanshinone IIA and dihydrotanshinone I using middle high-pressure preparation liquid phase method, and process includes: that the Radix Salviae Miltiorrhizae dregs of a decoction after (1) extracts water dry, pulverize, and obtains Radix Salviae Miltiorrhizae alcohol extract medicinal extract after extracting concentration with ethyl alcohol；(2) medicinal extract is extracted through petroleum ether, ethyl acetate respectively, takes ethyl acetate extract, concentration；(3) standby liquid phase is suppressed in using obtains total-tanshinone crude extract；(4) it is analyzed using UPLC, is then combined with tanshinone IIA and dihydrotanshinone I position (5) through the isolated tanshinone IIA of high pressure preparation liquid phase and dihydrotanshinone I monomeric compound.The present invention is to use middle high-pressure preparation solution phase joint technology for the first time; it can the efficiently and rapidly tanshinone IIA of preparative separation purification of high-purity and dihydrotanshinone I compound from the Radix Salviae Miltiorrhizae dregs of a decoction; waste resource is made full use of, there is important environmental protection meaning and economic value.
Description
Technical field
The invention belongs to Chinese medicinal plant chemical fields, and in particular to prepare liquid phase extraction purification Radix Salviae Miltiorrhizae using high pressure to a kind of
The preparation method of ketone IIA and dihydrotanshinone I monomeric compound.
Background technique
Radix Salviae Miltiorrhizae (Salvia miltiorrhiza Bge.) belongs to Lamiaceae plant, and main product is in Hebei, Anhui, Jiangsu, Sichuan
Etc. ground, first recorded in Shennong's Herbal, the effects of promoting blood circulation and removing obstruction in channels, cool blood to disappear carbuncle, stasis-dispelling and pain-killing, relieving restlessness and restlessness can be played.It is red
The chemical component of ginseng is divided into fat-soluble and water-soluble two parts, and tanshinone (tanshinone, Tsn) is fat-soluble rosin in Radix Salviae Miltiorrhizae
Alkane type diterpene-kind compound, also known as total-tanshinone have extensive pharmacological action, the highest attention by medical scholar.Rouge
The tanshinone compound of dissolubility has extensive pharmacological action and clinical application.Its chemical structure characteristic is that have o-quinone or right
Quinone structure, and it can be reduced to diphenol analog derivative, it is converted into quinone again after oxidation, plays the work of transmitting electronics in the process
With；And its interior metabolism product can also influence a variety of reactions of body, therefore have antitumor, antiatherosclerosis, drop
Low myocardial oxygen consumption and the Clinical pharmacological efficiencies such as antibacterial, anti-inflammatory, neuroprotection.
Danshen-containing injections mostly use traditional aqueous extraction-alcohol precipitation technology in production process, which makes in Radix Salviae Miltiorrhizae
Fat-soluble tanshinone component cannot be extracted and remain in the solid dregs of a decoction, cause the waste of resource.As world population is old
The trend in age, cardiovascular patient disease incidence rise, and the annual consumption of red rooted salvia is in growth trend year by year.2012,
China's danshen-containing injections market scale reaches nearly 15,700,000,000 yuan, 2013 annual nearly 19,000,000,000 yuan, occupy entire traditional Chinese medicine injection
The share in 35% or more market.However, generating a large amount of Radix Salviae Miltiorrhizae solid waste in danshen-containing injections preparation of the production process, lead
It causes Radix Salviae Miltiorrhizae resource material use efficiency low, while causing the serious waste and environmental pollution of natural resources of Chinese medicinal materials.Therefore, Salvia miltiorrhiza
Efficient utilize of resource substance has become inevitable development trend in slag.
Tanshinone component has antiatherosclerosis, reduction myocardial oxygen consumption and antibacterial, anti-oxidant, anti-inflammatory etc. a variety of
Bioactivity and application value, it is especially prominent especially in terms of anti-tumor activity.Tanshinone IIA has improvement coronary blood
Circulation, vascular endothelial cell reparation, antiatherosclerosis (AS) formation, resisting cardiac hypertrophy and prevention and treatment diabetic complication etc.
Effect；Dihydrotanshinone Ⅰ have significantly inhibit acetylcholine ester enzyme effect, by inhibit cell Proliferation, promote Apoptosis and
Differentiation is to play antitumor action and significant antibacterial activity etc..
The method of purifying tanshinone class compound mainly has alcohol extracting-water precipitating, macroporous resin purification, CO at present2Overcritical extraction
Take, chloroform extraction etc., using mesolow prepare chromatographic process separating-purifying fat soluble ingredient of red sage root, when can greatly shorten separation
Between, under the premise of not influencing separating degree, but separation preparation effect can be greatly improved with isolated a greater amount of compound
Rate.
The present invention relates to tanshinone monomeric compounds, include tanshinone IIA and dihydrotanshinone I, chemical structural formula
It is as follows:
With people to the bioactivity of the tanshinone compounds such as tanshinone IIA, dihydrotanshinone Ⅰ and Cryptotanshinone and
Development and application research is goed deep into, and has great scientific meaning to the purifying preparation of above-mentioned tanshinone monomeric compound and applies valence
Value.
Summary of the invention
Goal of the invention: the purpose of the present invention is the dregs of a decoction after making full use of danshen injections to extract, and provide a kind of using high
Suppress the preparation method of standby liquid phase extraction purification tanshinone IIA and dihydrotanshinone I monomeric compound.
Technical solution: in order to reach the above technical effect, technical scheme is as follows:
A method of tanshinone IIA and dihydrotanshinone I, including following step are prepared using middle high-pressure preparation liquid phase method
It is rapid:
Step A: it extracts: Salvia miltiorrhiza slag grinding and sieving being obtained into Salvia miltiorrhiza ground-slag end, ethanol solution is added, flows back
It extracts 2~3 times, 1~2 hour every time, combined extract was simultaneously concentrated to get Radix Salviae Miltiorrhizae alcohol extract medicinal extract；
Step B: abstraction impurity removal: the Radix Salviae Miltiorrhizae alcohol extract medicinal extract for taking step A to be prepared adds pure water to dissolve, uses petroleum respectively
Ether and ethyl acetate successively extract, and extract repeatedly three times, combined ethyl acetate position extracting solution, are concentrated under reduced pressure, obtain ethyl acetate
Position concentrate；
Step C: middle to suppress standby liquid phase purifying: to weigh the ethyl acetate extract concentrate that step B is prepared and silica gel is added
It stirs evenly, is then charged into middle pressure sample injector, selects petroleum ether and ethyl acetate for mobile phase, setting flow rate of mobile phase is 15
~20mL/min suppresses standby liquid phase in progress and purifies；
Step D: it using the middle compacting of UPLC analytical procedure C for liquid phase eluate, is then combined with tanshinone IIA and dihydro is red
Join the position ketone I:
Step E: high pressure preparation liquid phase purifying: the tanshinone IIA for taking step D to merge and dihydrotanshinone I position, concentration,
Methanol dissolution is added, is purified with the laggard high pressure preparation liquid phase of 0.22 μm of membrane filtration, selects methanol and ultrapure water for mobile phase, ladder
Degree elution purifies to obtain tanshinone IIA and dihydrotanshinone I through high pressure preparation liquid phase.
Preferably, above-described that tanshinone IIA and dihydrotanshinone are prepared using middle high-pressure preparation liquid phase method
The method of I, which is characterized in that the 8-12 times of ethyl alcohol for measuring 70-90% volumetric concentration is added in step A.
Preferably, above-described that tanshinone IIA and dihydrotanshinone are prepared using middle high-pressure preparation liquid phase method
The method of I, which is characterized in that the ethyl alcohol of 8 times of 90% volumetric concentrations of amount is added in step A, carries out refluxing extraction 2 times, 2 is small every time
When,
Preferably, above-described that tanshinone IIA and dihydrotanshinone are prepared using middle high-pressure preparation liquid phase method
The method of I suppresses gradient elution program in standby liquid phase in step C are as follows: and it be petroleum ether-B is mutually ethyl acetate that mobile phase, which is A phase,
Gradient elution: 0~5min, 95%A；5~12.4min, 95%~28%A；12.4~15.3min, 0%.
Preferably, above-described that tanshinone IIA and dihydrotanshinone are prepared using middle high-pressure preparation liquid phase method
The method of I, the chromatographic condition of the UPLC in step D are ACQU-ITY UPLCRBEH C18Chromatographic column；30 DEG C of column temperature；Flow velocity
0.4mL/min；2 μ L of sample volume, mobile phase: A phase is acetonitrile, and B phase is 0.1% aqueous formic acid, gradient elution: 0~1min,
70%A, 1~6min, 70%~80%A, 6~10min, 100%A.
Preferably, above-described that tanshinone IIA and dihydrotanshinone are prepared using middle high-pressure preparation liquid phase method
The method of I, step E high pressure prepare the liquid-phase condition of liquid phase: Waters Xbridge OBD C18Chromatographic column, 19mm × 150mm, 5
μm, mobile phase is that A phase is methanol-B Xiang Weishui, gradient elution: 0~5min, 75%A；5~15min, 75%~80%A；15~
35min, 90%A；35~45min, 100%A, Detection wavelength 285nm；Column temperature is room temperature；Flow velocity 15mL/min；500 μ of sample volume
L；Mass Spectrometry Conditions: ESI ion source, scanning mode are positive ion mode；Orifice potential: 50V, capillary voltage: 3kV, parent ion
Collision energy: 4V, fragment ion collision energy: 20-50V；Ion source temperature: 105 DEG C, desolvation temperature: 350 DEG C；Precipitation
Agent throughput: 700L/h, taper hole throughput: 50L/h；Mass scan range: 350-600Da, sweep time: 0.3s.
Preferably, above-described that tanshinone IIA and dihydrotanshinone are prepared using middle high-pressure preparation liquid phase method
The method of I, the Radix Salviae Miltiorrhizae dregs of a decoction described in step A be danshen-containing injections in process of production, the dregs of a decoction after being extracted using water.
The utility model has the advantages that provided by the invention prepare tanshinone IIA and dihydrotanshinone I using middle high-pressure preparation liquid phase method
Method, screen optimal purification step by many experiments, technological design is reasonable, suppresses standby liquid phase and height in particular by middle
It suppresses standby liquid phase to be combined, the Radix Salviae Miltiorrhizae dregs of a decoction yield that can isolate and purify to obtain purity is high from the Radix Salviae Miltiorrhizae dregs of a decoction is high, can make full use of useless
Resource is abandoned, turns waste into wealth, achieves extraordinary technical effect.
Detailed description of the invention
Fig. 1 is that tanshinone high pressure prepares chromatogram.
Fig. 2 is the chromatogram for the UPLC that high pressure prepares the dihydrotanshinone Ⅰ that liquid phase purifies.
Fig. 3 is the UPLC chromatogram that high pressure prepares the tanshinone IIA that liquid phase purifies.
Specific embodiment
Come the technical solution that the present invention will be described in detail, herein schematic implementation of the invention below in conjunction with specific embodiment
Example and explanation technical solution used to explain the present invention, but it is not as a limitation of the invention.
Embodiment 1
The method that tanshinone IIA and dihydrotanshinone I are prepared using middle high-pressure preparation liquid phase method, comprising the following steps:
Step A: the dregs of a decoction after Salvia miltiorrhiza Bge water is extracted, sieving obtains Danshen Root after dry, pulverize, and 8 times of 70 volumes of amount are added
The ethanol solution of concentration carries out refluxing extraction 2 times, and 2 hours every time, combined extract was simultaneously concentrated to get Radix Salviae Miltiorrhizae alcohol extract medicinal extract.
Step B: taking 100g Radix Salviae Miltiorrhizae alcohol extract medicinal extract, and 1000mL pure water is added to dissolve, and is extracted respectively with petroleum ether and ethyl acetate
It takes, extracts repeatedly three times, combined ethyl acetate position extracting solution is concentrated under reduced pressure, obtains ethyl acetate extract concentrate.
Step C: the silica gel of ethyl acetate extract concentrate one times of amount of addition to be separated is weighed in mortar, is ground
Mill.By sample be fitted into press sample injector in, make its be slowly added into keep sample plane level and it is smooth.Standby color is suppressed according to middle
The group reload request of spectrometer assembles chromatographic column.Selective flow is mutually petroleum ether and ethyl acetate.Setting flow rate of mobile phase is 20mL/
Min, gradient elution: 0~5min, 95%A；5~12.4min, 95%~28%A；12.4~15.3min, 0%.By mobile phase
It is fixed on collecting frame with collection vessel.After preparation is completed, profit column program is clicked.After moistening column completion, chromatography is clicked
Program, and eluent is collected through eluting with brown bottle, 45 bottles of eluent of collection, 50mL bottles every altogether.
Step D:UPLC analysis, segmentation merge.The chromatographic condition of UPLC is ACQU-ITY UPLCRBEH C18(2.1mm×
100mm, 1.7 μm) chromatographic column；30 DEG C of column temperature；Flow velocity 0.4mL/min；2 μ L of sample volume, mobile phase: -0.1% formic acid of acetonitrile (A)
Aqueous solution (B), gradient elution (0~1min, 70%A, 1~6min, 70%~80%A, 6~10min, 100%A).UPLC points
After analysis, six sections are broadly divided into, merges eluent, concentration.And high pressure preparation purifying is carried out to No. 5 collection liquids.Centering compacting is standby to wash
De- liquid concentration, is recovered under reduced pressure solvent, and appropriate 70% methanol dissolution prepares liquid phase with the laggard high pressure of 0.22 μm of membrane filtration.
Step E: high pressure prepares chromatographic condition: liquid-phase condition: Waters Xbridge OBD C18Chromatographic column (19mm ×
150mm, 5 μm), mobile phase is methanol (A)-water (B), gradient elution (0~5min, 75%A；5~15min, 75%~80%A；
15~35min, 90%A；35~45min, 100%A；).Detection wavelength 285nm；Column temperature is room temperature；Flow velocity 15mL/min；Sample introduction
Measure 500 μ L.Mass Spectrometry Conditions: ESI ion source, scanning mode are positive ion mode；Orifice potential: 50V, capillary voltage: 3kV,
Parent ion collision energy: 4V, fragment ion collision energy: 20-50V；Ion source temperature: 105 DEG C, desolvation temperature: 350
℃；Desolventizing gas flow: 700L/h, taper hole throughput: 50L/h；Mass scan range: 350-600Da, sweep time: 0.3s.
It is eluted, collects eluent.After UPLC analysis, 3 sections are broadly divided into, merges eluent, is concentrated, as shown in Figure 1.First and third section
Respectively compound dihydrotanshinone Ⅰ, tanshinone IIA, content can obtain crystal through recrystallization 90% or more.It takes appropriate
Crystal 1 and crystal 3 carry out purity detecting into UPLC.Tanshinone IIA purity is 98.8%, as shown in Fig. 2, dihydrotanshinone Ⅰ is pure
Degree is as shown in Figure 3 for 98.2%.
Embodiment 2
The method that tanshinone IIA and dihydrotanshinone I are prepared using middle high-pressure preparation liquid phase method, comprising the following steps:
Step A: the dregs of a decoction after Salvia miltiorrhiza Bge water is extracted, dry, Radix Salviae Miltiorrhizae grinding and sieving obtains Danshen Root, 10 times of amounts is added
The ethanol solution of 80% volumetric concentration carries out refluxing extraction 2 times, and 2 hours every time, combined extract was simultaneously concentrated to get Radix Salviae Miltiorrhizae alcohol extracting
Object medicinal extract.
Step B: taking 300g Radix Salviae Miltiorrhizae dregs of a decoction alcohol extracting thing medicinal extract, and 3000mL pure water is added to dissolve, and uses petroleum ether and acetic acid respectively
Ethyl ester extraction, extracts 3 times, combined ethyl acetate position extracting solution repeatedly, is concentrated under reduced pressure, obtains ethyl acetate extract concentrate.
Step C: the silica gel of sample ethyl acetate extract concentrate one times of amount of addition to be separated is weighed in mortar, is carried out
Grinding.By sample be fitted into press sample injector in, make its be slowly added into keep sample plane level and it is smooth.It is standby according to middle compacting
Chromatographic group of reload request assembles chromatographic column.Selective flow is mutually petroleum ether and ethyl acetate.Flow rate of mobile phase, which is arranged, is
20mL/min, gradient elution: 0~5min, 95%A；5~12.4min, 95%~28%A；12.4~15.3min, 0%.It will stream
It is dynamic to be mutually fixed on collecting frame with collection vessel.After preparation is completed, profit column program is clicked.After moistening column completion, click
Chromatography Program, and eluent is collected through eluting with brown bottle, 90 bottles of eluent of collection, 50mL bottles every altogether.
Step D:UPLC analysis, segmentation merge, and the chromatographic condition of UPLC is ACQU-ITY UPLCRBEH C18(2.1mm×
100mm, 1.7 μm) chromatographic column；30 DEG C of column temperature；Flow velocity 0.4mL/min；2 μ L of sample volume, mobile phase: -0.1% formic acid of acetonitrile (A)
Aqueous solution (B), gradient elution (0~1min, 70%A, 1~6min, 70%~80%A, 6~10min, 100%A).UPLC points
After analysis, six sections are broadly divided into, merges eluent, concentration.And high pressure preparation purifying is carried out to No. 5 collection liquids.Centering compacting is standby to wash
De- liquid concentration, is recovered under reduced pressure solvent, and appropriate 70% methanol dissolution prepares liquid phase with the laggard high pressure of 0.22 μm of membrane filtration.
Step E: high pressure prepares chromatographic condition: liquid-phase condition: Waters Xbridge OBD C18Chromatographic column (19mm ×
150mm, 5 μm), mobile phase is methanol (A)-water (B), gradient elution (0~5min, 75%A；5~15min, 75%~80%A；
15~35min, 90%A；35~45min, 100%A；).Detection wavelength 285nm；Column temperature is room temperature；Flow velocity 15mL/min；Sample introduction
Measure 500 μ L.Mass Spectrometry Conditions: ESI ion source, scanning mode are positive ion mode；Orifice potential: 50V, capillary voltage: 3kV,
Parent ion collision energy: 4V, fragment ion collision energy: 20-50V；Ion source temperature: 105 DEG C, desolvation temperature: 350
℃；Desolventizing gas flow: 700L/h, taper hole throughput: 50L/h；Mass scan range: 350-600Da, sweep time: 0.3s.
It is eluted, collects eluent.After UPLC analysis, 3 sections are broadly divided into, merges eluent, concentration.First and third section is respectively chemical combination
Object dihydrotanshinone Ⅰ, tanshinone IIA, content can obtain crystal through recrystallization 90% or more.Take appropriate crystal 1 and crystal
3, purity detecting is carried out into UPLC.Tanshinone IIA purity is 98.9%, and dihydrotanshinone Ⅰ purity is 98.8%.
Embodiment 3
The method that tanshinone IIA and dihydrotanshinone I are prepared using middle high-pressure preparation liquid phase method, comprising the following steps:
Step A: the dregs of a decoction after Salvia miltiorrhiza Bge water is extracted, dry, Radix Salviae Miltiorrhizae grinding and sieving obtains Danshen Root, 12 times of amounts is added
The ethanol solution of 90% volumetric concentration carries out refluxing extraction 2 times, and 2 hours every time, combined extract was simultaneously concentrated to get Radix Salviae Miltiorrhizae alcohol extracting
Object medicinal extract.
Step B: taking 200g Radix Salviae Miltiorrhizae dregs of a decoction alcohol extracting thing medicinal extract, and 2000mL pure water is added to dissolve, and uses petroleum ether and acetic acid respectively
Ethyl ester extraction, extracts three times, combined ethyl acetate position extracting solution repeatedly, is concentrated under reduced pressure, obtains ethyl acetate extract concentrate.
Step C: the silica gel of sample ethyl acetate extract concentrate one times of amount of addition to be separated is weighed in mortar, is carried out
Grinding.By sample be fitted into press sample injector in, make its be slowly added into keep sample plane level and it is smooth.It is standby according to middle compacting
Chromatographic group of reload request assembles chromatographic column.Selective flow is mutually petroleum ether and ethyl acetate.Flow rate of mobile phase, which is arranged, is
The ratio and chromatogram of 20mL/min, petroleum ether and ethyl acetate are shown in annex.Mobile phase and collection vessel are fixed on collecting frame
On.After preparation is completed, profit column program is clicked.After moistening column completion, Chromatography Program is clicked, and with brown bottle to eluent
It is collected through eluting, 45 bottles of eluent of collection, 50mL bottles every altogether.
Step D:UPLC analysis segmentation merges, and the chromatographic condition of UPLC is ACQU-ITY UPLCRBEH C18(2.1mm×
100mm, 1.7 μm) chromatographic column；30 DEG C of column temperature；Flow velocity 0.4mL/min；2 μ L of sample volume, mobile phase: -0.1% formic acid of acetonitrile (A)
Aqueous solution (B), gradient elution (0~1min, 70%A, 1~6min, 70%~80%A, 6~10min, 100%A).UPLC points
After analysis, six sections are broadly divided into, merges eluent, concentration.And high pressure preparation purifying is carried out to No. 5 collection liquids.Centering compacting is standby to wash
De- liquid concentration, is recovered under reduced pressure solvent, and appropriate 70% methanol dissolution prepares liquid phase with the laggard high pressure of 0.22 μm of membrane filtration.
Step E: high pressure prepares chromatographic condition: liquid-phase condition: Waters Xbridge OBD C18Chromatographic column (19mm ×
150mm, 5 μm), mobile phase is methanol (A)-water (B), gradient elution (0~5min, 75%A；5~15min, 75%~80%A；
15~35min, 90%A；35~45min, 100%A；).Detection wavelength 285nm；Column temperature is room temperature；Flow velocity 15mL/min；Sample introduction
Measure 500 μ L.Mass Spectrometry Conditions: ESI ion source, scanning mode are positive ion mode；Orifice potential: 50V, capillary voltage: 3kV,
Parent ion collision energy: 4V, fragment ion collision energy: 20-50V；Ion source temperature: 105 DEG C, desolvation temperature: 350
℃；Desolventizing gas flow: 700L/h, taper hole throughput: 50L/h；Mass scan range: 350-600Da, sweep time: 0.3s.
It is eluted, collects eluent.After UPLC analysis, 3 sections are broadly divided into, merges eluent, concentration.First and third section is respectively chemical combination
Object dihydrotanshinone Ⅰ, tanshinone IIA, content can obtain crystal through recrystallization 90% or more.Take appropriate crystal 1 and crystal
3, purity detecting is carried out into UPLC, tanshinone IIA purity is 98.6%, and dihydrotanshinone Ⅰ purity is 98.1%.
Claims (4)
1. a kind of method for preparing tanshinone IIA and dihydrotanshinone I using middle high-pressure preparation liquid phase method, which is characterized in that packet
Include following steps:
Step A: it extracts: Salvia miltiorrhiza slag grinding and sieving being obtained into Salvia miltiorrhiza ground-slag end, ethanol solution is added, carries out refluxing extraction
2~3 times, 1~2 hour every time, combined extract was simultaneously concentrated to get Radix Salviae Miltiorrhizae alcohol extract medicinal extract；
Step B: abstraction impurity removal: the Radix Salviae Miltiorrhizae alcohol extract medicinal extract for taking step A to be prepared adds pure water to dissolve, respectively with petroleum ether and
Ethyl acetate successively extracts, and extracts repeatedly three times, combined ethyl acetate position extracting solution, is concentrated under reduced pressure, obtains ethyl acetate extract
Concentrate；
Step C: middle to suppress standby liquid phase purifying: to weigh the ethyl acetate extract concentrate that step B is prepared and silica gel stirring is added
Uniformly, be then charged into middle pressure sample injector, select petroleum ether and ethyl acetate for mobile phase, setting flow rate of mobile phase be 15~
20mL/min suppresses standby liquid phase in progress and purifies；
Step D: using the middle compacting of UPLC analytical procedure C for liquid phase eluate, tanshinone IIA and dihydrotanshinone are then combined with
The position I:
Step E: high pressure preparation liquid phase purifying: the tanshinone IIA for taking step D to merge and dihydrotanshinone I position are concentrated, and are added
Methanol dissolution is purified with the laggard high pressure preparation liquid phase of 0.22 μm of membrane filtration, selects methanol and ultrapure water for mobile phase, gradient is washed
It is de-, tanshinone IIA and dihydrotanshinone I are purified to obtain through high pressure preparation liquid phase；
Step C, it is middle to suppress gradient elution program in standby liquid phase are as follows: it be petroleum ether-B is mutually ethyl acetate, gradient that mobile phase, which is A phase,
Elution: 0~5min, 95%A；5~12.4min, 95%~28%A；12.4~15.3min, A0%；
The chromatographic condition of UPLC in step D is ACQU-ITY UPLCRBEH C18Chromatographic column；30 DEG C of column temperature；Flow velocity 0.4mL/
min；2 μ L of sample volume, mobile phase: A phase is acetonitrile, and B phase is 0.1% aqueous formic acid, gradient elution: 0~1min, 70%A, 1
~6min, 70%~80%A, 6~10min, 100%A；
The liquid-phase condition of step E high pressure preparation liquid phase: WatersXbridge OBD C18Chromatographic column, 19mm × 150mm, 5 μm, stream
It is methanol-B Xiang Weishui, gradient elution: 0~5min, 75%A that dynamic phase, which is A phase,；5~15min, 75%~80%A；15~
35min, 90%A；35~45min, 100%A, Detection wavelength 285nm；Column temperature is room temperature；Flow velocity 15mL/min；500 μ of sample volume
L；Mass Spectrometry Conditions: ESI ion source, scanning mode are positive ion mode；Orifice potential: 50V, capillary voltage: 3kV, parent ion
Collision energy: 4V, fragment ion collision energy: 20-50V；Ion source temperature: 105 DEG C, desolvation temperature: 350 DEG C；Precipitation
Agent throughput: 700L/h, taper hole throughput: 50L/h；Mass scan range: 350-600Da, sweep time: 0.3s.
2. the side according to claim 1 for preparing tanshinone IIA and dihydrotanshinone I using middle high-pressure preparation liquid phase method
Method, which is characterized in that the 8-12 times of ethyl alcohol for measuring 70-90% volumetric concentration is added in step A.
3. the side according to claim 1 for preparing tanshinone IIA and dihydrotanshinone I using middle high-pressure preparation liquid phase method
Method, which is characterized in that the ethyl alcohol of 8 times of 90% volumetric concentrations of amount is added in step A, carries out refluxing extraction 2 times, every time 2 hours.
4. the side according to claim 1 for preparing tanshinone IIA and dihydrotanshinone I using middle high-pressure preparation liquid phase method
Method, which is characterized in that
The Radix Salviae Miltiorrhizae dregs of a decoction described in step A be danshen-containing injections in process of production, using water extract after the dregs of a decoction.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710026412.6A CN106831936B (en) | 2017-01-13 | 2017-01-13 | The method that tanshinone IIA and dihydrotanshinone I are prepared using middle high-pressure preparation liquid phase method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710026412.6A CN106831936B (en) | 2017-01-13 | 2017-01-13 | The method that tanshinone IIA and dihydrotanshinone I are prepared using middle high-pressure preparation liquid phase method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106831936A CN106831936A (en) | 2017-06-13 |
| CN106831936B true CN106831936B (en) | 2019-04-19 |
Family
ID=59124934
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710026412.6A Active CN106831936B (en) | 2017-01-13 | 2017-01-13 | The method that tanshinone IIA and dihydrotanshinone I are prepared using middle high-pressure preparation liquid phase method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106831936B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112439004A (en) * | 2019-09-05 | 2021-03-05 | 上海中医药大学 | Salvia miltiorrhiza extract and preparation method and application thereof |
| CN111559996A (en) * | 2020-06-15 | 2020-08-21 | 劲牌有限公司 | A method for preparing salvianolic acid B, cryptotanshinone, tanshinone I and tanshinone IIA from Saviae Miltiorrhizae radix |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2646240A1 (en) * | 2006-03-16 | 2007-09-20 | Xuemin Shi | Combination therapy for treatment of patients with neurological disorders and cerebral infarction |
| CN102127037A (en) * | 2011-01-11 | 2011-07-20 | 上海交通大学 | Tanshinone compounds and applications thereof |
| CN102167721B (en) * | 2011-03-24 | 2012-10-10 | 聊城大学 | Method for extracting and purifying tanshinone monomeric compounds from red sage root |
| CN104497096A (en) * | 2015-01-07 | 2015-04-08 | 南京中医药大学 | Method for extracting and preparing tanshinone compounds from dregs of radix salviae miltiorrhizae by virtue of saccharification pretreatment method |
-
2017
- 2017-01-13 CN CN201710026412.6A patent/CN106831936B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN106831936A (en) | 2017-06-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105585471A (en) | Extraction method of active ingredients of isodon amethystoides | |
| CN103739586A (en) | Method for extracting diterpenoid compounds from Blumea aromatic DC. | |
| CN106831936B (en) | The method that tanshinone IIA and dihydrotanshinone I are prepared using middle high-pressure preparation liquid phase method | |
| CN109879919B (en) | Method for separating and preparing three flavonoid glycosides from spina date seeds | |
| CN101104010A (en) | Extracting and purifying technology for haw flavonoids substances | |
| CN104829474B (en) | Method for preparing glycine betaine chemical reference substances from boxthorn leaves | |
| CN102219685B (en) | A kind of preparation method of danshen root salvianolic acid A | |
| CN102391350A (en) | Method for purifying polygalasaponin F | |
| CN106632577B (en) | A kind of Ursane triterpenoid saponin and its preparation process in masson pine branch | |
| CN102670680A (en) | Method for extracting total saponins from bark of acanthopanax brachypus harms | |
| CN101084967A (en) | Extracting and purifying technology for safflower flavonoids | |
| CN110551137B (en) | Method for extracting and purifying glabridin and application of glabridin in cosmetics | |
| CN103030678B (en) | Method for preparing rare ginsenoside from malonyl ginsenoside | |
| CN108379316A (en) | A kind of extracting method of ginsenoside | |
| CN108003217A (en) | A kind of method that peroxy-ergosterol is extracted from Cordyceps cicadae | |
| CN103772478A (en) | Single-column cyclic separation system for preparing high-purity tanshinone compounds and method thereof | |
| CN101328198B (en) | Extraction and separation method of syringin | |
| CN106420871A (en) | Preparation method of Panax notoginseng saponins rich in Fe and Fd and monomers Fe and Fd thereof | |
| CN104402949B (en) | A kind of method that separation simultaneously prepares Syringin and oleuropein from lilac | |
| CN114644608B (en) | Fisetin with urate transporter 1 inhibitory activity, and preparation method and application thereof | |
| CN111304272A (en) | Method for preparing rare ginsenoside by converting panax notoginseng stem and leaf saponin through fusarium verticillii | |
| CN105294793A (en) | Separation method for naringin in aizoon stonecrop herb | |
| CN109293726A (en) | Diol type ginsenoside extract and preparation method thereof | |
| CN110204588B (en) | Method for preparing astragaloside | |
| CN103665070B (en) | The extraction and separation technology of a kind of limonin glycosides and production technology |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |