CN104926896A - Method and system for extracting and separating Vincenin-1 in Desmodium styracifolium - Google Patents
Method and system for extracting and separating Vincenin-1 in Desmodium styracifolium Download PDFInfo
- Publication number
- CN104926896A CN104926896A CN201510347049.9A CN201510347049A CN104926896A CN 104926896 A CN104926896 A CN 104926896A CN 201510347049 A CN201510347049 A CN 201510347049A CN 104926896 A CN104926896 A CN 104926896A
- Authority
- CN
- China
- Prior art keywords
- ethanol
- separate part
- chromatographic separation
- alcohol
- carry out
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention provides a method for extracting and separating Vincenin-1. The method includes the steps of subjecting Desmodium styracifolium to reflux extraction via ethyl alcohol to obtain Desmodium styracifolium extract; subjecting the extract to multistage chromatographic separation via alcohol systems, to be specific, subjecting the extract to first chromatographic separation via the first alcohol system to obtain a first separate part, subjecting the first separate part to second chromatographic separation via the second alcohol system to obtain a second separate part, and subjecting the second separate part to third chromatographic separation via the third alcohol system to so as to obtain the Vincenin-1. The invention further provides a system implementing the method. The process of separating and extracting the Vincenin-1 by the method and the system from Desmodium styracifolium is simple to operate, production time is short, product yield is high, purity is high, and the Vincenin-1 is up to above 95% in purity.
Description
Technical field
The invention belongs to technical field of phytochemistry, concrete, the present invention relates to the method and system of a kind of extraction and isolation Vicenin-1 from Snowbell-leaf Tickelover Herb.
Background technology
Snowbell-leaf Tickelover Herb is pulse family beggar-ticks plant Desmodium styracifolium (Osbeck) Merr., and its medicinal part is dry aerial parts, and main chemical compositions is the compounds such as flavones, saponin(e, polysaccharide, alkaloid.There is removing damp-heat, effect of inducing diuresis for treating stranguria syndrome.For heat pouring, Sha Lin, urolithiasis, difficulty and pain in micturition, oedema oliguria, red, the lithangiuria of jaundice urine.
Vicenin-1, also referred to as 6-C-wood sugar-8-C-glucose apiolin, is yellow powder; Molecular formula: C
26h
28o
14; Molecular weight: 564; Fig. 1 shows its chemical structural formula.Vicenin-1 is present in Snowbell-leaf Tickelover Herb medicinal material, differentiates and the reference substance of assay as Snowbell-leaf Tickelover Herb, and Snowbell-leaf Tickelover Herb and relevant pharmaceutical formulations such as the quality control of Snowbell-leaf Tickelover Herb total flavone capsule thereof are used.The method and system of existing extraction and isolation Vicenin-1 urgently improves.
Summary of the invention
The present invention is intended to solve at least to a certain extent the problems of the prior art one of at least or provide a kind of business to select.For this reason, one object of the present invention is the method and system proposing a kind of extraction and isolation Vicenin-1 from Snowbell-leaf Tickelover Herb, utilize the Vicenin-1 that this extraction and separation method and system obtain, purity is high, is conducive to differentiating as Snowbell-leaf Tickelover Herb and the reference substance of assay and the quality control as Snowbell-leaf Tickelover Herb and relevant pharmaceutical formulations thereof.
According to an aspect of of the present present invention, the invention provides a kind of method of extraction and isolation Vicenin-1, the method comprises the following steps: utilize ethanol to carry out refluxing extraction to Snowbell-leaf Tickelover Herb, obtains Herba Desmodii Styracifolii extract; Alcohol system is utilized to carry out multistage chromatography separation to described extract, comprising, the first alcohol system is utilized to carry out the first chromatographic separation to described extract, obtain the first separate part, diol system is utilized to carry out the second chromatographic separation to described first separate part, obtain the second separate part, utilize triol system to carry out tertiary color spectrum to described second separate part and be separated, to obtain described Vicenin-1.
The method of the present invention, utilize multistage chromatography to be separated, comprise combination and utilize macroporous resin purification technology, the standby and gel chromatography technology of middle compacting, adopt alcohol-water system, separation and Extraction Vicenin-1 from Herba Desmodii Styracifolii extract, this technological operation is simple, and the production time is short, product yield is high, purity is high, can obtain the Vicenin-1 that purity reaches more than 95%, and the Vicenin-1 of acquisition can be used as assay chemical reference substance.The Parameter Conditions of this processing step and each step, contriver considers, repeatedly the combination of Adjustment Tests macroporous resin chromatogram, anti-phase middle pressure chromatogram and gel chromatography and the impact of respective elution requirement on the separation and purification effect of the Vicenin-1 in Snowbell-leaf Tickelover Herb thereof are decided, this extraction purification process does not relate to the relatively strong organic solvent of toxicity, and extraction and isolation operating process and extract are all safe and reliable.
Fig. 2 shows the flow chart of steps of the method for compound shown in the extraction and isolation Fig. 1 in one embodiment of the present of invention.According to embodiments of the invention, the invention described above Vicenin-1 extraction and separation method on the one hand, can also have following technical characteristic one of at least:
According to one embodiment of present invention, the described ethanol that utilizes carries out refluxing extraction to Snowbell-leaf Tickelover Herb, obtain Herba Desmodii Styracifolii extract, comprising: utilize the ethanol of concentration 50-95% to carry out twice refluxing extraction to described Snowbell-leaf Tickelover Herb, merge the extracting solution of twice refluxing extraction gained, obtain united extraction liquid, remove the ethanol in described united extraction liquid, and/or the ethanol removed in described united extraction liquid and ion, and optional, carry out drying, to obtain described extract.Twice concentration is that 50-95% alcohol reflux obtains extract, is that contriver gropes to determine through test of many times, can makes full use of raw material and also can save time, and is beneficial to large-scale production preparation; Remove the ethanol in extracting solution and/or ion, be beneficial to and get rid of them to the interference of later separation purifying.
According to one embodiment of present invention, described twice refluxing extraction, wherein the weight ratio of ethanol once and Snowbell-leaf Tickelover Herb is 12:1, the time of refluxing extraction is 1.5h-3h, that optional is 2h, the ethanol of another time and the weight ratio of Snowbell-leaf Tickelover Herb are 10:1, and the time of refluxing extraction is 1h-2h, and that optional is 1.5h.During backflow, concentration is ratio and the return time of 50-95% ethanol and Snowbell-leaf Tickelover Herb, is that contriver determines through test of many times detection, raw material can be made to obtain high extraction.
According to some embodiments of the present invention, described first alcohol system is ethanol-water system, and the concentration of the ethanol in this ethanol-water system is 18-25%, and preferably, the concentration of the ethanol in described ethanol-water system is 20%, and described first chromatogram is macroporous resin.The concentration of the ethanol Vicenin-1 ratio that to be contriver in conjunction with follow-up chromatographic separation condition, test of many times detect in the peak eluted in this alcohol-water eluent system, determines through repeatedly adjusting and optimizing.
According to some embodiments of the present invention, described diol system is ethanol-water system, the concentration of the ethanol in this ethanol-water system is 6-30%, described second chromatogram is reverse chromatograms, and preferably, the reverse chromatograms of selection is reverse middle pressure chromatogram, better, select C18 post, its post pressure is 1-10bar, and elution flow rate is 5 ~ 30ml/min.Alcohol concn in this alcohol-water eluent system is the ratio of contriver in conjunction with target component in the first separate part, and the first alcohol-water eluent system in chromatographic separation, test of many times optimization is determined, be beneficial in simple operations with in the short period of time, obtain the Vicenin-1 that yield is high, purity is high.
According to one embodiment of present invention, the described diol system that utilizes carries out the second chromatographic separation to the first separate part, obtain the second separate part, comprise: the ethanol-water system utilizing alcohol concn to be a carries out isocratic elution 25-40min to described first separate part, then using alcohol concn instead is [a, b] ethanol-water system gradient elution 180-250min is carried out to described first separate part, collect to obtain described second separate part, wherein, the span of a is the span of 6-12%, b is 15-30%.First utilize the ethanol-water system isocratic elution of a concentration, then carry out gradient elution, separation and Extraction efficiency can be improved; The alcohol concn of isocratic elution and gradient elution or concentration range, and elution time, be the ratio etc. that contriver considers target component in yield, time, the complexity of separate part of recovery, separate part, test of many times is determined.
As shown in Figure 3, according to another embodiment of the invention, the described diol system that utilizes carries out the second chromatographic separation to the first separate part, obtain the second separate part, comprise: the ethanol-water system utilizing alcohol concn to be c carries out isocratic elution 30min to described first separate part, then use the ethanol-water system that alcohol concn is [c, d] instead and gradient elution 200min is carried out to described first separate part, collect and obtain elementary second separate part; The ethanol-water system utilizing alcohol concn to be e carries out isocratic elution 30min to described elementary second separate part, then using alcohol concn instead is [e, f] ethanol-water system gradient elution 200min is carried out to described elementary second separate part, collect and obtain described second separate part, wherein, the span of c and e is the span of 6-12%, d and f is 15-30%, c>e, d>f.Front and back twice, all first utilize the ethanol-water system isocratic elution of a concentration, then carry out gradient elution, can improve the efficiency of separation and Extraction Vicenin-1; Before and after the isocratic elution of twice and the alcohol concn of gradient elution or concentration range, respective elution time, speed, it is the ratio etc. that contriver takes target component in the condition of two inferior degree-gradient elutions, yield, time, the complexity of separate part of recovery, separate part into consideration, test of many times optimization is determined, be beneficial in simple operations with in the short period of time, obtain the Vicenin-1 that yield is high, purity is high.In a preferred embodiment of the present invention, c=10%, e=8%, d=30%, f=15%, can make the final product purity obtained reach more than 95%.
According to another embodiment of the invention, described triol system is pure methyl alcohol, and described tertiary color spectrum is gel chromatography.In one embodiment of the invention, selected gel chromatography is sephadex chromatography Sephadex G-75.Be beneficial to the purity improving target compound further.
According to another aspect of the present invention, the invention provides the system of a kind of extraction and isolation Vicenin-1, this system can in order to implement the Vicenin-1 extraction and separation method of the invention described above on the one hand or in any embodiment, this system comprises: refluxing extraction device, in order to utilize ethanol to carry out refluxing extraction to Snowbell-leaf Tickelover Herb, obtain Herba Desmodii Styracifolii extract; Chromatographic separation device, for utilizing alcohol system to carry out multistage chromatography separation to described extract, it comprises, first chromatographic separation unit, is connected with described refluxing extraction device, carries out the first chromatographic separation for utilizing the first alcohol system to described extract, obtain the first separate part
Second chromatographic separation unit, be connected with described first chromatographic separation unit, for utilizing diol system, the second chromatographic separation is carried out to described first separate part, obtain the second separate part, tertiary color spectrum separating unit, be connected with described second chromatographic separation unit, for utilizing triol system, tertiary color spectrum carried out to described second separate part and be separated, to obtain described Vicenin-1.Fig. 4 shows the structure of the extraction and isolation system 1000 in one embodiment of the present of invention, comprise refluxing extraction device 100 and chromatographic separation device 200, chromatographic separation device comprises the first chromatographic separation unit 202, second chromatographic separation unit 204 and tertiary color spectrum separating unit 206.The above-mentioned technical characteristic of Vicenin-1 extraction and separation method to one aspect of the present invention and the description of advantage, be suitable for the system of this one side of the present invention equally, do not repeat them here.It will be understood by those skilled in the art that this system can also comprise other device, sub-device or functional unit to implement above-mentioned corresponding embodiment.
Additional aspect of the present invention and advantage will part provide in the following description, and part will become obvious from the following description, or be recognized by practice of the present invention.
Accompanying drawing explanation
Above-mentioned and/or additional aspect of the present invention and advantage will become obvious and easy understand from accompanying drawing below combining to the description of embodiment, wherein:
Fig. 1 shows the chemical structural formula of Vicenin-1;
Fig. 2 shows the steps flow chart of the method for compound shown in the extraction and isolation Fig. 1 in one embodiment of the present of invention;
Fig. 3 shows the steps flow chart of the method for compound shown in the extraction and isolation Fig. 1 in one embodiment of the present of invention;
Fig. 4 shows the structural representation of the system of compound shown in the extraction and isolation Fig. 1 in one embodiment of the present of invention;
Fig. 5 shows the schema of the method for compound shown in the extraction and isolation Fig. 1 in one embodiment of the present of invention;
Fig. 6 shows the ESI-MS mass spectrum of the extraction purification product in one embodiment of the invention;
Fig. 7 shows the H of the extraction purification product in one embodiment of the invention
1-NMR spectrogram; And
Fig. 8 shows the C of the extraction purification product in one embodiment of the invention
13-NMR spectrogram.
Embodiment
Be described below in detail embodiments of the invention, the example of described embodiment is shown in the drawings, and wherein same or similar label represents same or similar element or has element that is identical or similar functions from start to finish.Being exemplary below by the embodiment be described with reference to the drawings, only for explaining the present invention, and can not limitation of the present invention being interpreted as.
It should be noted that, term " first ", " second " only for describing object, and can not be interpreted as instruction or hint relative importance or imply the quantity indicating indicated technical characteristic.Thus, be limited with " first ", the feature of " second " can express or impliedly comprise one or more these features.Further, in describing the invention, except as otherwise noted, the implication of " multiple " is two or more.
Below in conjunction with embodiment, the solution of the present invention is made an explanation.It will be understood to those of skill in the art that the following examples only for illustration of the present invention, and should not be considered as limiting scope of the present invention.Unreceipted concrete technology or condition in embodiment, according to the technology described by the document in this area or condition or carry out according to product description.Agents useful for same or the unreceipted production firm person of instrument, being can by the conventional products of commercial acquisition.
Embodiment 1
Fig. 5 shows the general flow of the inventive method, comprises following:
1, Feedstock treating: select leguminous plants Snowbell-leaf Tickelover Herb 45000-50000 weight part, be cut into 5-10cm segment, tap water washes silt, drains, and feeds intake.
2, extraction using alcohol: 80% alcohol reflux twice, first time 12 times amount 2h, second time 10 times amount 1.5h; Merge twice alcohol extract, reclaim ethanol extremely without alcohol taste.
3, concentrating under reduced pressure: extract 5 times of volumes that concentrated solution is diluted with water to medicinal material weight; 200 order filter-cloth filterings; Obtain upper prop liquid; To fill post with the clean product resin that 95% Ethanol Treatment is crossed in advance, purified water is replaced; Upper prop liquid pump is squeezed in post, absorption, coutroi velocity 0.5-1 times BV/h, after absorption, leave standstill 2h; Rinse by purified water, elution flow rate 3-4BV/h, elution amount 10BV; Use 60% ethanol elution again, elution flow rate 2-3BV/h, elution amount 5BV; 60% elutriant merges, and squeezes into concentration tank, and reclaim ethanol extremely without alcohol taste, 50-70 DEG C of drying to vacuum drying oven, obtains extensively golden bulk drug 300-400 weight part.
4, multistage chromatography is separated
(1) by wide golden bulk drug 300-400 weight part AB-8 macroporous resin chromatographic separation, i.e. chromatographic separation I, be specially: use ethanol-water mixed solvent 20:80,30:70,40:60,60:40 gradient elution, each gradient is flushed to without color, merges each stream part and obtains 20%, 30%, 40%, 60% totally four positions.
(2) the anti-phase middle pressure chromatographic separation of 20% position ODS chromatographic separation I obtained, i.e. chromatographic separation II, be specially: with alcohol-water, condition is 10% 30min such as degree such as grade, and 10-30% gradient 200min wash-out, according to color atlas peak sequence, portioning is collected, every 100-150 parts by volume is collected a, and collect 40-50 stream part altogether, high performance liquid chromatography inspection merges same stream part after knowing.
(3) 10-29 that chromatographic separation II obtains is flowed part merging, use the anti-phase middle pressure chromatographic separation of ODS further, i.e. chromatographic separation III, is specially: with alcohol-water, condition is 8% 30min such as degree such as grade, 8-15% gradient 200min wash-out, according to color atlas peak sequence, portioning is collected, and every 100-150 parts by volume is collected a, collect 35-45 stream part altogether, high performance liquid chromatography inspection merges same stream part after knowing.
(4) 25-35 that chromatographic separation III obtains is flowed part merging, being evaporated to a small amount of precipitation can stop.Be placed to precipitation pale yellow precipitate further, filter and obtain Vicenin-1 crude product.Finally be separated (SephadexG-75) with sephadex chromatography, i.e. chromatographic separation IV, compound shown in Fig. 1 is added in methanol-water (volume ratio is 1:1), ultrasonic dissolution, sampling volume is 10 milliliters, rushes post with pure methyl alcohol, portioning is collected, every 5 parts by volume are collected a, and collect 20-30 stream part altogether, high performance liquid chromatography inspection merges same stream part after knowing; The 9-20 that chromatographic separation IV obtains flows part merging, and recycling design obtains yellow powder.
Wherein, (2) and (3) i.e. chromatographic separation II and chromatographic separation III utilizes the anti-phase middle pressure chromatogram of ODS, corresponding above-mentioned the second alleged chromatographic separation.
5, Structural Identification
Through electrospray ionization mass spectrometry (ESI-MS) and spectrography (H
1-NMR and C
13-NMR) identify, in conjunction with ESI-MS, H obtaining yellow powder
1-NMR and C
13-NMR spectrogram information infers that the molecular formula of this compound is C
26h
28o
14determine that the above-mentioned yellow powder that extraction and isolation goes out from Snowbell-leaf Tickelover Herb is Vicenin-1, through HPLC purity test, purity reaches more than 95%, differentiate and the reference substance of assay as Snowbell-leaf Tickelover Herb, and Snowbell-leaf Tickelover Herb and relevant pharmaceutical formulations such as the quality control of Snowbell-leaf Tickelover Herb total flavone capsule thereof are used.
Fig. 6 is the mass spectrum that Vicenin-1 carries out electrospray ionization mass spectrometry, ESI-MS m/z:565 [M+H]
+, the molecular weight of each composition in display yellow powder.
Fig. 7 shows the H of Vicenin-1
1-NMR spectrogram, H
1-NMR (DMSO, 400HZ) δ: 13.76 (5-OH), 8.01 (2H, d, J=8.4Hz, H-2 ', 6 '), 6.92 (2H, d, J=8.4Hz, H-3 ', 5 '), 6.78 (1H, s, H-3).
Fig. 8 shows the C of Vicenin-1
13-NMR spectrogram, C
13-NMR (DMSO, 100HZ) δ: 163.7 (C-2), 102.5 (C-3), 182.2 (C-4), 161.1 (C-5), 108.1 (C-6), 159.2 (C-7), 103.4 (C-8), 155.2 (C-9), 102.5 (C-10), 121.5 (C-1 '), 128.9 (C-2 ' 6 '), 115.9 (C-3 ' 5 '), 161.3 (C-4 '), 74.9 (C-1 " of Xyl), 70.3 (C-2 " of Xyl), 77.8 (C-3 " of Xyl), 69.9 (C-4 " of Xyl), 69.6 (C-5 " of Xyl), 72.3 (C-1 " ' of Glc), 71.3 (C-2 " ' of Glc), 78.8 (C-3 " ' of Glc), 70.5 (C-4 " ' of Glc), 81.8 (C-5 " ' of Glc), 60.6 (C-6 " ' of Glc).Wherein said Xyl represents wood sugar, and Glc represents glucose.
Embodiment 2
Select leguminous plants Snowbell-leaf Tickelover Herb 45000-50000 weight part, be cut into 5-10cm segment, tap water washes silt, drains, and feeds intake.80% alcohol reflux twice, first time 12 times amount 2h, second time 10 times amount 1.5h; Merge twice alcohol extract, reclaim ethanol extremely without alcohol taste.Extract 5 times of volumes that concentrated solution is diluted with water to medicinal material weight; 200 order filter-cloth filterings; Obtain upper prop liquid; To fill post with the clean product resin that 95% Ethanol Treatment is crossed in advance, purified water is replaced; Upper prop liquid pump is squeezed in post, absorption, coutroi velocity 0.5-1 times BV/h, after absorption, leave standstill 2h; Rinse by purified water, elution flow rate 3-4BV/h, elution amount 10BV; Use 60% ethanol elution again, elution flow rate 2-3BV/h, elution amount 5BV; 60% elutriant merges, and squeezes into concentration tank, and reclaim ethanol extremely without alcohol taste, 50-70 DEG C of drying to vacuum drying oven, obtains extensively golden bulk drug 300-400 weight part.
By wide golden bulk drug 300-400 weight part AB-8 macroporous resin chromatographic separation, i.e. chromatographic separation I, ethanol-water mixed solvent 25:75,35:65,45:55,65:35 gradient elution, each gradient is flushed to without color, merges each stream part and obtains 25%, 35%, 45%, 65% totally four positions; The anti-phase middle pressure chromatographic separation of 25% position ODS that chromatogram I obtains, i.e. chromatographic separation II, with alcohol-water, condition is 8% 30min such as degree such as grade, and 8-30% gradient 200min wash-out, according to color atlas peak sequence, portioning is collected, every 100-150 parts by volume is collected a, and collect 40-50 stream part altogether, high performance liquid chromatography inspection merges same stream part after knowing; The 15-30 that chromatographic separation II obtains flows part merging, use the anti-phase middle pressure chromatographic separation of ODS further, i.e. chromatographic separation III, with alcohol-water, condition is 6% 30min such as degree such as grade, 6-15% gradient 200min wash-out, according to color atlas peak sequence, portioning is collected, and every 100-150 parts by volume is collected a, collect 35-45 stream part altogether, high performance liquid chromatography inspection merges same stream part after knowing; The 20-35 that chromatographic separation III obtains flows part merging, is recycled in solvent and has a small amount of precipitation to stop.Be placed to precipitation pale yellow precipitate further, filter and obtain Vicenin-1 crude product.Finally be separated (SephadexG-75), i.e. chromatographic separation IV with sephadex chromatography, rush post with pure methyl alcohol, portioning is collected, and every 5 parts by volume are collected a, and collect 20-30 stream part altogether, high performance liquid chromatography inspection merges same stream part after knowing; The 10-20 that chromatographic separation IV obtains flows part merging, and recycling design obtains yellow powder.Utilize electrospray ionization mass spectrometry (ESI-MS) and spectrography (H
1-NMR and C
13-NMR) identify obtaining yellow powder, identify that yellow powder is Vicenin-1, purity 88.7%.
Embodiment 3
Select leguminous plants Snowbell-leaf Tickelover Herb 45000-50000 weight part, be cut into 5-10cm segment, tap water washes silt, drains, and feeds intake.80% alcohol reflux twice, first time 12 times amount 2h, second time 10 times amount 1.5h; Merge twice alcohol extract, reclaim ethanol extremely without alcohol taste.Extract 5 times of volumes that concentrated solution is diluted with water to medicinal material weight; 200 order filter-cloth filterings; Obtain upper prop liquid; To fill post with the clean product resin that 95% Ethanol Treatment is crossed in advance, purified water is replaced; Upper prop liquid pump is squeezed in post, absorption, coutroi velocity 0.5-1 times BV/h, after absorption, leave standstill 2h; Rinse by purified water, elution flow rate 3-4BV/h, elution amount 10BV; Use 60% ethanol elution again, elution flow rate 2-3BV/h, elution amount 5BV; 60% elutriant merges, and squeezes into concentration tank, and reclaim ethanol extremely without alcohol taste, 50-70 DEG C of drying to vacuum drying oven, obtains extensively golden bulk drug 300-400 weight part.
By wide golden bulk drug 300-400 weight part AB-8 macroporous resin chromatographic separation, i.e. chromatographic separation I, ethanol-water mixed solvent 20:80,30:70,40:60,60:40 gradient elution, each gradient is flushed to without color, merges each stream part and obtains 20%, 30%, 40%, 60%, totally four positions; The anti-phase middle pressure chromatographic separation of 20% position ODS that chromatogram I obtains, i.e. chromatographic separation II, with alcohol-water, condition is 12% 30min such as degree such as grade, and 12-30% gradient 200min wash-out, according to color atlas peak sequence, portioning is collected, every 100-150 parts by volume is collected a, and collect 40-50 stream part altogether, high performance liquid chromatography inspection merges same stream part after knowing; The 15-35 that chromatographic separation II obtains flows part merging, use the anti-phase middle pressure chromatographic separation of ODS further, i.e. chromatographic separation III, with alcohol-water, condition is 10% 30min such as degree such as grade, 10-15% gradient 200min wash-out, according to color atlas peak sequence, portioning is collected, and every 100-150 parts by volume is collected a, collect 35-45 stream part altogether, high performance liquid chromatography inspection merges same stream part after knowing; The 25-40 that chromatographic separation III obtains flows part merging, is recycled in solvent and has a small amount of precipitation to stop.Be placed to precipitation pale yellow precipitate further, filter and obtain Vicenin-1 crude product.Finally be separated (Sephadex G-75), i.e. chromatographic separation IV with sephadex chromatography, rush post with pure methyl alcohol, portioning is collected, and every 5 parts by volume are collected a, and collect 20-30 stream part altogether, high performance liquid chromatography inspection merges same stream part after knowing; The 10-25 that chromatographic separation IV obtains flows part merging, and recycling design obtains yellow powder.Identify obtaining yellow powder through electrospray ionization mass spectrometry (ESI-MS) and spectrography (H1-NMR and C13-NMR), yellow powder is Vicenin-1, purity 90.1%.
The yellow powder Vicenin-1 that the processing condition of embodiment 2 and 3 are produced, purity is significantly lower than the Vicenin-1 of embodiment 1 separation and Extraction.
In the description of this specification sheets, specific features, structure, material or feature that the description of reference term " embodiment ", " some embodiments ", " example ", " concrete example " or " some examples " etc. means to describe in conjunction with this embodiment or example are contained at least one embodiment of the present invention or example.In this manual, identical embodiment or example are not necessarily referred to the schematic representation of above-mentioned term.And the specific features of description, structure, material or feature can combine in an appropriate manner in any one or more embodiment or example.
Although illustrate and describe embodiments of the invention, those having ordinary skill in the art will appreciate that: can carry out multiple change, amendment, replacement and modification to these embodiments when not departing from principle of the present invention and aim, scope of the present invention is by claim and equivalents thereof.
Claims (10)
1. a method of extraction and isolation Vicenin-1, is characterized in that, comprises the following steps:
Utilize ethanol to carry out refluxing extraction to Snowbell-leaf Tickelover Herb, obtain Herba Desmodii Styracifolii extract;
Alcohol system is utilized to carry out multistage chromatography separation to described extract, comprising,
Utilize the first alcohol system to carry out the first chromatographic separation to described extract, obtain the first separate part,
Utilize diol system to carry out the second chromatographic separation to described first separate part, obtain the second separate part,
Utilize triol system to carry out tertiary color spectrum to described second separate part to be separated, to obtain described Vicenin-1.
2. method according to claim 1, is characterized in that, the described ethanol that utilizes carries out refluxing extraction to Snowbell-leaf Tickelover Herb, obtains Herba Desmodii Styracifolii extract, comprising:
Utilize the ethanol of concentration 50-95% to carry out twice refluxing extraction to described Snowbell-leaf Tickelover Herb, merge the extracting solution of twice refluxing extraction gained, obtain united extraction liquid,
Remove the ethanol in described united extraction liquid, and/or the ethanol removed in described united extraction liquid and ion, and optional, carry out drying, to obtain described extract.
3. method according to claim 2, is characterized in that, described twice refluxing extraction, wherein
Ethanol once and the weight ratio of Snowbell-leaf Tickelover Herb are 12:1, and the time of refluxing extraction is 1.5h-3h, and that optional is 2h,
The ethanol of another time and the weight ratio of Snowbell-leaf Tickelover Herb are 10:1, and the time of refluxing extraction is 1h-2h, and that optional is 1.5h.
4. method according to claim 1, is characterized in that, described first alcohol system is ethanol-water system, and the concentration of the ethanol in described ethanol-water system is 18-25%,
Optional, the concentration of the ethanol in described ethanol-water system is 20%,
Described first chromatogram is macroporous resin.
5. method according to claim 1, is characterized in that, described diol system is ethanol-water system,
The concentration of the ethanol in described ethanol-water system is 6-30%,
Described second chromatogram is reverse chromatograms.
6. method according to claim 5, is characterized in that, described reverse chromatograms is reverse middle pressure chromatogram,
Optional, described oppositely middle pressure chromatogram is C18 post, and its post pressure is 1-10bar, and elution flow rate is 5 ~ 30ml/min.
7. method according to claim 5, is characterized in that, the described diol system that utilizes carries out the second chromatographic separation to the first separate part, obtains the second separate part, comprising:
The ethanol-water system utilizing alcohol concn to be a carries out isocratic elution 25-40min to described first separate part, then using alcohol concn instead is [a, b] ethanol-water system gradient elution 180-250min is carried out to described first separate part, collect to obtain described second separate part, wherein
The span of a is the span of 6-12%, b is 15-30%.
8. method according to claim 5, is characterized in that, the described diol system that utilizes carries out the second chromatographic separation to the first separate part, obtains the second separate part, comprising:
The ethanol-water system utilizing alcohol concn to be c carries out isocratic elution 30min to described first separate part, then using alcohol concn instead is [c, d] ethanol-water system gradient elution 200min is carried out to described first separate part, collect obtain elementary second separate part
The ethanol-water system utilizing alcohol concn to be e carries out isocratic elution 30min to described elementary second separate part, then use the ethanol-water system that alcohol concn is [e, f] instead and gradient elution 200min is carried out to described elementary second separate part, collect and obtain described second separate part, wherein
The span of c and e is the span of 6-12%, d and f is 15-30%, c>e, d>f.
9. method according to claim 1, is characterized in that, described triol system is pure methyl alcohol, and described tertiary color spectrum is gel chromatography.
10. a system of extraction and isolation Vicenin-1, is characterized in that, comprising:
Refluxing extraction device, in order to utilize ethanol to carry out refluxing extraction to Snowbell-leaf Tickelover Herb, obtains Herba Desmodii Styracifolii extract;
Chromatographic separation device, for utilizing alcohol system to carry out multistage chromatography separation to described extract, it comprises,
First chromatographic separation unit, is connected with described refluxing extraction device, for utilizing the first alcohol system to carry out the first chromatographic separation to described extract, obtains the first separate part,
Second chromatographic separation unit, being connected with described first chromatographic separation unit, for utilizing diol system to carry out the second chromatographic separation to described first separate part, obtaining the second separate part,
Tertiary color spectrum separating unit, is connected with described second chromatographic separation unit, carries out tertiary color spectrum be separated, to obtain described Vicenin-1 for utilizing triol system to described second separate part.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510347049.9A CN104926896A (en) | 2015-06-19 | 2015-06-19 | Method and system for extracting and separating Vincenin-1 in Desmodium styracifolium |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510347049.9A CN104926896A (en) | 2015-06-19 | 2015-06-19 | Method and system for extracting and separating Vincenin-1 in Desmodium styracifolium |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104926896A true CN104926896A (en) | 2015-09-23 |
Family
ID=54114340
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510347049.9A Pending CN104926896A (en) | 2015-06-19 | 2015-06-19 | Method and system for extracting and separating Vincenin-1 in Desmodium styracifolium |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104926896A (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012150486A1 (en) * | 2011-05-02 | 2012-11-08 | Indus Biotech Private Limited | A composition for treating autoimmune disorders and methods thereof |
-
2015
- 2015-06-19 CN CN201510347049.9A patent/CN104926896A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012150486A1 (en) * | 2011-05-02 | 2012-11-08 | Indus Biotech Private Limited | A composition for treating autoimmune disorders and methods thereof |
Non-Patent Citations (7)
| Title |
|---|
| HARDER, LENE H.，ET AL.: "A new flavone O-glycoside and other constituents from wheat leaves", 《ZEITSCHRIFT FUER NATURFORSCHUNG》 * |
| MAGDY MOSTAFA DESOKY MOHAMMED，ET AL.: "The cytotoxic activity of Linum grandiflorum leaves", 《EUROPEAN JOURNAL OF CHEMISTRY》 * |
| PARK, HAE-SUK，ET AL.: "Antioxidant flavone glycosides from the leaves of Sasa borealis", 《ARCHIVES OF PHARMACAL RESEARCH》 * |
| ZHU, TIAN MI，ET AL.: "A new flavone glycoside from Selaginella moellendorffii Hieron", 《CHINESE CHEMICAL LETTERS》 * |
| 周子力: "广金钱草中三种黄酮碳苷的制备与含量测定", 《西南交通大学研究生学位论文》 * |
| 张蕾，等: "大孔吸附树脂富集纯化广金钱草总黄酮的工艺研究", 《中草药》 * |
| 王国光，等: "反相高效液相色谱法制备广金钱草黄酮对照品", 《中成药》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102838644B (en) | Production method for extracting sweet tea glucoside from sweet tea leaves | |
| CN101302548B (en) | Preparation of icaritin | |
| CN102166235B (en) | Extraction and purification method of saikosaponin | |
| CN105399656A (en) | Isobenzazole alkaloid compound, and preparation method and applications thereof | |
| CN104926897A (en) | Method and system for extracting and separating novel Schaftoside in Desmodium styracifolium | |
| CN101255182B (en) | Extraction separating method for bupleurum root saponin b2 | |
| CN102617669A (en) | Method for separating and purifying mangiferin from mango pericarp | |
| CN104910216B (en) | It is a kind of with preparing liquid phase method while obtaining the separation method of a variety of epimedium flavones | |
| CN102391350A (en) | Method for purifying polygalasaponin F | |
| CN105061182A (en) | Method for extracting, separating and purifying emodin and physcion from polygonum cuspidatum | |
| CN105017273A (en) | Method for extracting, separating and purifying psoralen and isopsoralen from fructus psoraleae | |
| CN104945355A (en) | Method and system for extracting and separating dihydro-phaseic acid from desmodium styracifolium | |
| CN102260307A (en) | Method for preparing specnuezhenide | |
| CN104945391B (en) | The method and system of extraction separation Schaftoside from Desmodium styracifolium | |
| CN101955504A (en) | Method for preparing specnuezhenide, nuezhenoside and oleanolic acid by high-speed countercurrent chromatography | |
| CN105131007A (en) | Method for extracting, separating and purifying imperatorin and cnidium lactone from fructus cnidii | |
| CN103479751A (en) | Method for combined extraction of tritepenoidic acid, polyphenols and polysaccharides in loquat flower | |
| CN103254165A (en) | Preparation method of atractylenolide II | |
| CN105085498A (en) | Method and system for extraction and separation of isovitexin from Desmodium styracifolium | |
| CN102617674A (en) | Preparation method of scopolin monomer in anisodus tanguticus root | |
| CN102311466A (en) | Method for extracting phenylethanoid glycoside active components from semenplantaginis | |
| CN104926896A (en) | Method and system for extracting and separating Vincenin-1 in Desmodium styracifolium | |
| CN105037338A (en) | Method and system for extracting and separating luteolin-6-C-beta-D-glucopyranose-8-C-beta-xylopyranoside | |
| CN103965276B (en) | The method of fast separating and purifying monomeric compound from Lindley Eupatorium Herb | |
| CN105001190A (en) | Method and system for extracting and separating Vicenin-2 from desmodium styracifolium |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150923 |
|
| RJ01 | Rejection of invention patent application after publication |