CN106967029A - One kind prepares isofraxidin method from Chinese medicinal material of sarcandra glaber - Google Patents
One kind prepares isofraxidin method from Chinese medicinal material of sarcandra glaber Download PDFInfo
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- CN106967029A CN106967029A CN201610028246.9A CN201610028246A CN106967029A CN 106967029 A CN106967029 A CN 106967029A CN 201610028246 A CN201610028246 A CN 201610028246A CN 106967029 A CN106967029 A CN 106967029A
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- isofraxidin
- medicinal material
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/06—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2
- C07D311/08—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring
- C07D311/18—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring substituted otherwise than in position 3 or 7
Abstract
Isofraxidin method is prepared from Chinese medicinal material of sarcandra glaber the present invention relates to one kind, it is using Glabrous Sarcandra Herb complete stool medicine materical crude slice as raw material, a large amount of unrelated compositions are removed by water heating extracting, liquid-liquid extraction method, obtain isofraxidin crude product enrichment positions, finely separated through silica gel column chromatography, gel column chromatography depigmentation, anti-phase ODS column chromatographys again, recrystallizing and refining, you can obtain purity be more than 99% isofraxidin sterling.Present invention process flow operations are simple, and production cost is low, and products obtained therefrom purity is high.
Description
Technical field
The present invention relates to a kind of isofraxidin preparation method, belong to medicinal active ingredient extractive technique field.
Background technology
Isofraxidin is natural cumarin constituents, with preferable anti-inflammatory, analgesia, anti-infectious function in vivo
（Xiaofeng Niu etc., International Immunopharmacology. 2015. 24：432–439; Ling Liu
Deng Immunobiology, 2015,220: 406–413；Xiaofeng Niu etc., International
Immunopharmacology. 2012. 14：164–171）, it is Chinese medicine Glabrous Sarcandra Herb, wilsonii medicinal material characteristic chemical constituent, and make
Recorded for quality of medicinal material Con trolling index composition in 2015 editions Chinese Pharmacopoeias（Chinese Pharmacopoeia, 2015 editions, I portions, the 206th, 224
Page）, for the control of Glabrous Sarcandra Herb, wilsonii medicinal material and its quality of the pharmaceutical preparations.The preparation method of current document report isofraxidin has greatly
Hole resin chromatography（He Hangzhen, Jiangxi College of Traditional Chinese Medicine journal, 2007,19（4）：45-46）, high-speed countercurrent chromatography（Jiang Yongnan
Deng Chinese medicine, 2008,31 (6)：913-915）.But the cumarin crude product position Zhong Yi Qin that macroreticular resin partition method is prepared
Skin pyridine content is only 2.59%, and uses the valuable cost height of high-speed counter-current instrument, consumption of organic solvent big, and prepares enrichment positions
Still it is crude product, isofraxidin yield only has 3%.Other isofraxidin process of preparing for having applied for patent of invention are included from thorn
Alcohol extracting, sour water solution, the extraction of 1,1- dichloroethanes obtain isofraxidin, but are also only crude product in slender acanthopanax rhizome（Zu YuanGang,
A kind of method that isofraxidin is prepared in rhizome from wilsonii, the patent No.：201010184243.7）.So far, also without one
Kind of simple possible, the isofraxidin monomer preparation technology for preparing relative inexpensiveness disclose report.
The content of the invention
The present invention overcomes prior art not enough, and there is provided a kind of simple to operate, with low cost and products obtained therefrom purity is high
Isofraxidin preparation method.
The implementation of the object of the invention is：
A. Chinese medicinal material of sarcandra glaber medicine materical crude slice is taken, 2 times are taken with the heating of 5-15 times of raw-material weight water, every time 30 120 min, merging is carried
Filtrate is taken, is filtered, filtrate is standby；
B. filtrate is concentrated into proper volume, cyclohexane-ethyl acetate（1:1, volume ratio）Equal-volume extraction 1-4 times；
C. cyclohexane-ethyl acetate extraction phase vacuum-concentrcted, organic solvent redissolution, 1-3 times of silica gel dry method mixes sample, through 5-20
Times silica gel column chromatography, petroleum ether：Acetone（15:8, volume ratio）Elution, tlc analysis merges and contains isofraxidin flow point；
D. methanol redissolves merges flow point rich in isofraxidin, and through gel column chromatography, methanol elutes depigmentation, then through ODS column chromatographys,
MeOH-H2O is eluted, and obtains purity more than 85% crude product isofraxidin, methanol-water recrystallization produces purity more than 99% sterling.
Compared with prior art, the advantage of the invention is that：The present invention extracts low with raw medicinal material wide material sources, price
It is honest and clean；Water heating extracting method is used to Chinese medicinal material of sarcandra glaber medicine materical crude slice, pigment impurity in all grass herb extract solution is greatly reduced；
Hydrophilic, lipophilic feature is had concurrently according to isofraxidin, extracted using cyclohexane-ethyl acetate mixed organic solvents, can maximum journey
Degree reduces other water soluble ingredient coextractions, reduction pigment interference, so as to reduce the follow-up cumbersome unrelated impurity of removal and pigment
Lock out operation；Liquid-liquid extraction organic solvent is recyclable to be recycled；The present invention is simple to operate, is set without using expensive instrument
It is standby, so that sample preparation cost lowers significantly.
Brief description of the drawings
Fig. 1 is isofraxidin UV abosrption spectrograms.
Fig. 2 is isofraxidin electron spray（ESI）Positive ion mode high resolution mass spectrum figure.
Fig. 3 is isofraxidin proton magnetic（1H-NMR, 500 MHz, DMSO-d6）Spectrogram.
Fig. 4 is isofraxidin carbon nuclear-magnetism（1H-NMR, 125 MHz, DMSO-d6）Spectrogram.
Fig. 5 is Glabrous Sarcandra Herb aqueous extract HPLC chromatogram.
Fig. 6 is Glabrous Sarcandra Herb aqueous extract cyclohexane-ethyl acetate extraction part HPLC chromatogram.
Fig. 7 is that separation prepares isofraxidin HPLC chromatogram.
Embodiment
Embodiment
1. isofraxidin extracts separation
Take the Kg of Glabrous Sarcandra Herb herb medicinal material 12（About 3 cm/ sections）, put in 100 L Chinese medicine extracting tanks, 84 L water infiltrations added for the first time
0.5 hour, 100 DEG C of heating decocted 2h, and second plus 42 L water, 100 DEG C of heating decoct 1.5 h, gauze filtration is concentrated into 7 L
（Equivalent to the g/ml of medicinal material concentration about 1.7）, cyclohexane-ethyl acetate（1:1）Equal-volume extraction 4 times, combining extraction liquid, concentration
Dry, acetate-methanol dissolution extraction reclaims soluble fraction（44 g）, 100 g column chromatography silica gels（100~200 mesh）Uniformly
Sample is mixed, room temperature flings to solvent, 700 g column chromatography silica gels（200~300 mesh）Dry column-packing, petroleum ether II：Acetone（15:8）Gradient
2 column volumes are eluted, every 500 ml is a Fraction collection, and concentration and recovery organic phase obtains each subflow point, tlc analysis, oil
Ether II（60-90 DEG C of boiling point）：Acetone（1:1）Expansion, petroleum ether II：Acetone（15:8）6-16 bottles are eluted in the nm of uviol lamp 365
Lower and isofraxidin reference substance same position shows sky blue fluorescence spot.Merge and stayed point containing isofraxidin, through gel（Sephadex
LH-20）Column chromatography, methanol elution, tlc analysis merges containing isofraxidin subflow point, then through ODS column chromatographys, methanol-water
（20:80®30:70）Elution, 30% methanol-eluted fractions obtain isofraxidin crude product (2.9 g), recrystallize, produce through methanol-water
Isofraxidin sterling.
2. isofraxidin Structural Identification
White powder（MeOH）, UV λmax(MeOH) nm: 342 nm（Accompanying drawing 1）; HR-ESI-MS+：m/z 223.05983
[M+H]++（Calcd for C11H11O5 +, -1.2 ppm）（Accompanying drawing 2）.1H-NMR (500 MHz, DMSO-d6) δ:
7.89 (1H, d, J = 9.5 Hz, H-4), 6.22 (1H, d, J = 9.5 Hz, H-3), 7.01 (1H, s, H-
5), 3.82 (3H, s, 6-OCH3), 3.81 (3H, s, 8-OCH3)。13C-NMR (125 MHz, DMSO-d6) δ:
160.7(C-2), 146.2(C-6), 145.3(C-4), 144.9(C-9), 143.5(C-7), 135.2(C-8), 112.1
(C-3), 104.9(C-5), 110.4(C-10), 61.0(OCH3-8), 56.6(OCH3- 5) accompanying drawing 3-4, is seen.
[Xu Xudong, Hu Xiaoru, Yuan Jingquan, Yang Junshan wait cumarin chemistry in Chloranthus glabers to above nuclear magnetic data with document
Composition Study CHINA JOURNAL OF CHINESE MATERIA MEDICAs .2008,33 (8), 900-902] report it is basically identical.
3. isofraxidin high-efficient liquid phase analysis
3.1 qualitative analysis
Instrument：The efficient analysis liquid phases of Waters 2695, with 2998 PAD diode permutation detectors.Main chromatographic condition：Chromatogram
Post：XBridgeTMC18 (4.6´250 mm, 5μm)；Column temperature：40℃；The ml/min of flow velocity 0.8；Detection wavelength：330 nm;Stream
Dynamic phase：Methanol (A) -0.1% formic acid water (B) condition of gradient elution is 0-10 min, 20 22%A, 10-30 min, 20 37%
A, 30-45 min, 37®42%A；Sampling volume：20 μl.
Need testing solution is prepared and analyzed：Medicinal material aqueous extract equivalent to Chinese medicinal material of sarcandra glaber 0.2g/ml, phase are prepared respectively
When the isofraxidin reference substance that the organic solvent extraction in the mg/ml of extract 10 partly and from Glabrous Sarcandra Herb separates preparation is supplied
Test sample solution, sample introduction HPLC analyses, chromatogram is shown in accompanying drawing 5-7.
3.2 quantitative analysis
Detection wavelength：341 nm;Mobile phase：The phosphoric acid water of acetonitrile -0.1%（20:80）；Other conditions are with 3.1 qualitative analyses.
Reference substance solution is prepared and standard curve is set up：Isofraxidin standard items（Middle inspection institute, batch 110837-201507,
Content is in terms of 99.2%）, precision weighs 1.03,1.07 mg to ten a ten thousandth analytical precision balances respectively, pipettes 1ml methanol, surpasses
Sound dissolves, and produces mother liquor storage liquid, wherein concentration is that 1.03 mg/ml mother liquors dilute 2,4,6,8,16,32,64 respectively,
128,256 times, the μ l of sample introduction 20 are set up for standard curve, and concentration is that 1.07 mg/ml dilute 8 times for Quality Control, through instrument
Analysis software processing, linear equation is y=6.54e+004X -7.08e+004, and r=0.9999, the range of linearity is 3.99-
510.88 μg/ml。
Need testing solution is prepared and assay：It is parallel respectively to prepare 3 parts of QC control liquids, isofraxidin crude product, different barks of ash
Pyridine recrystallization, Glabrous Sarcandra Herb herb Aqueous extracts（Equivalent to the g/ml of medicinal material 0.2）And Glabrous Sarcandra Herb herb water extraction hexamethylene-acetic acid second
Ester（1:1）Extract part（Equivalent to the extraction g/ml of medicinal extract 0.01）, HPLC sample introductions analysis, calculate content, measure and the results are shown in Table
One.
The of table one prepares isofraxidin and sample measurement result
| Sample | Quality-control sample | Isofraxidin crude product | Isofraxidin is crystallized | Aqueous extracts | Extract |
| Content (%) | 98.6±1.0 | 86.5%±0 | 105.5±0.4 | 0.03±0 | 2.8±0.1 |
3.3 interpretation of result
It was found from table one, isofraxidin purity can be used for assay up to 105% after crystallization.
It was found from accompanying drawing 5-7, under same Detection wavelength, except retention time is that 16.0 min are different in Glabrous Sarcandra Herb water extract
Bark of ash pyridine target component, also other interference compositions, and in Glabrous Sarcandra Herb water extract cyclohexane-ethyl acetate（1:1）Extraction portion
Divide, only one principal component of isofraxidin, it is 93 times of content in medicinal material further to measure the content in its extract（Table one）, adopt
It is with the obvious advantage with cyclohexane-ethyl acetate liquid-liquid extraction method.
This technological process is converted into medicinal material and prepares isofraxidin yield about 0.025%, and preparation yield is higher, so that sample
Cost is prepared significantly to lower.
The foregoing is only presently preferred embodiments of the present invention, be not intended to limit the invention, it is all the present invention spirit and
Any modification, equal replacement and improvement for being made within principle etc., all should fall within the scope and spirit of the invention.
Claims (5)
1. one kind prepares isofraxidin method from Chinese medicinal material of sarcandra glaber, it is characterised in that step is as follows：Chinese medicinal material of sarcandra glaber medicine materical crude slice is taken,
Water heating extracting, filtration, merging filtrate, concentration, cyclohexane-ethyl acetate mixed organic solvents extraction, combining extraction liquid is dense
Contracting, silica gel dry method is mixed sample, eluted through silica gel column chromatography, thin layer detection, merges and is rich in isofraxidin eluent flow point, then through ODS
Column chromatography, gel filtration chromatography, recrystallization method can prepare high-purity isofraxidin.
2. one kind according to claim 1 prepares isofraxidin method from Chinese medicinal material of sarcandra glaber, it is characterised in that：Heating is carried
It is 5-15 times of raw material weight to take aqueous solvent.
3. one kind according to claim 1 prepares isofraxidin method from Chinese medicinal material of sarcandra glaber, it is characterised in that：Extraction is used
Organic phase is cyclohexane-ethyl acetate mixed organic solvents.
4. one kind according to claim 1 prepares isofraxidin method from Chinese medicinal material of sarcandra glaber, it is characterised in that：Silicagel column
Chromatographic elution solvent is petroleum ether II- acetone systems, and ratio is 15: 8.
5. one kind according to claim 1 prepares isofraxidin method from Chinese medicinal material of sarcandra glaber, it is characterised in that：Column chromatography
Pattern is open or mesolow column chromatography.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610028246.9A CN106967029A (en) | 2016-01-14 | 2016-01-14 | One kind prepares isofraxidin method from Chinese medicinal material of sarcandra glaber |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113759030A (en) * | 2021-04-29 | 2021-12-07 | 北京康仁堂药业有限公司 | Thin-layer identification method for glabrous sarcandra herb and glabrous sarcandra herb formula preparation |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101574381A (en) * | 2009-02-02 | 2009-11-11 | 江中药业股份有限公司 | Extractive of effective part of chloranthus glaber and preparation method thereof |
| CN101665479A (en) * | 2009-09-22 | 2010-03-10 | 三明华健生物工程有限公司 | Technology for synchronously extracting isofraxidin and flavonoids compounds from sarcandra glabra and applications thereof |
| CN102633760A (en) * | 2012-03-20 | 2012-08-15 | 浙江维康药业有限公司 | Isofraxidin crystalline compound and glabrous sarcandra herb dispersible tablets and dropping pills containing isofraxidin crystalline compound |
| CN103494806A (en) * | 2013-09-12 | 2014-01-08 | 中山大学 | Application and preparation method of benzo alpha-pyrone compounds |
| CN103623017A (en) * | 2013-08-22 | 2014-03-12 | 广西医科大学 | Sarcandra extract and application thereof |
-
2016
- 2016-01-14 CN CN201610028246.9A patent/CN106967029A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101574381A (en) * | 2009-02-02 | 2009-11-11 | 江中药业股份有限公司 | Extractive of effective part of chloranthus glaber and preparation method thereof |
| CN101665479A (en) * | 2009-09-22 | 2010-03-10 | 三明华健生物工程有限公司 | Technology for synchronously extracting isofraxidin and flavonoids compounds from sarcandra glabra and applications thereof |
| CN102633760A (en) * | 2012-03-20 | 2012-08-15 | 浙江维康药业有限公司 | Isofraxidin crystalline compound and glabrous sarcandra herb dispersible tablets and dropping pills containing isofraxidin crystalline compound |
| CN103623017A (en) * | 2013-08-22 | 2014-03-12 | 广西医科大学 | Sarcandra extract and application thereof |
| CN103494806A (en) * | 2013-09-12 | 2014-01-08 | 中山大学 | Application and preparation method of benzo alpha-pyrone compounds |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113759030A (en) * | 2021-04-29 | 2021-12-07 | 北京康仁堂药业有限公司 | Thin-layer identification method for glabrous sarcandra herb and glabrous sarcandra herb formula preparation |
| CN113759030B (en) * | 2021-04-29 | 2023-03-17 | 北京康仁堂药业有限公司 | Thin-layer identification method for glabrous sarcandra herb and glabrous sarcandra herb formula preparation |
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