CN103800397B - The technique of macroporous resin purification Herba Ephedrae total alkaloids - Google Patents
The technique of macroporous resin purification Herba Ephedrae total alkaloids Download PDFInfo
- Publication number
- CN103800397B CN103800397B CN201410047475.6A CN201410047475A CN103800397B CN 103800397 B CN103800397 B CN 103800397B CN 201410047475 A CN201410047475 A CN 201410047475A CN 103800397 B CN103800397 B CN 103800397B
- Authority
- CN
- China
- Prior art keywords
- herba ephedrae
- total alkaloids
- ethanol
- eluant
- technique
- 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
Abstract
The invention provides a kind of technique of macroporous resin purification Herba Ephedrae total alkaloids, comprise the following steps：Three kinds of Contents of Main Components measure, the pretreatment of macroreticular resin, loadings, wash-outs in the preparation of Chinese ephedra extract solution, the assay of Herba Ephedrae total alkaloids, Chinese ephedra, there is provided a kind of optimised process for purifying Herba Ephedrae total alkaloids, it is easy to operate, yield is high, is that further galenic pharmacy and pharmacodynamic study are offered reference.
Description
Technical field
The invention belongs to the field of Chinese medicines, specifically a kind of technique of macroporous resin purification Herba Ephedrae total alkaloids.
Background technology
Chinese ephedraEphedrae HerbaIt is EphedraceaeEphedraceaPlant ephedra sinicaEphedra sinica Stapf, in
Chinese ephedraEphedra intermedia Schrenk et C.A.MeyAnd ephedra equisetinaEphedra equisetina BgeIt is dry
Dry herbaceous stem stem, be《Chinese Pharmacopoeia》The traditional Chinese medicine for recording.Modern study shows that the TA in Chinese ephedra has in excitement
Pivot, antitussive and antiasthmatic, lax tracheae, hypotensive, influence tissue anti-oxidation function, reduction blood sugar[It is that it plays medicine Deng pharmacological action
The active component of effect.
However, the research that isolates and purifies of Herba Ephedrae total alkaloids is rarely reported, only using resin to single group in Chinese ephedra
Point --- ephedrine hydrochloride carries out the investigation of purifying process, not can be shown that the purifying situation to Herba Ephedrae total alkaloids.
The content of the invention
In order to solve the above-mentioned technical problem, the invention provides a kind of technique of macroporous resin purification Herba Ephedrae total alkaloids.
A kind of technique of macroporous resin purification Herba Ephedrae total alkaloids, it is characterised in that comprise the following steps：
1）Precision weighs Chinese ephedra fine powder in right amount, 8 times of 0.5% aqueous hydrochloric acid solutions of amount of addition, 60 DEG C, 250W ultrasonic extraction 1h,
Suction filtration, required concentration is concentrated under reduced pressure into Rotary Evaporators；
2）Precision draws appropriate Chinese ephedra need testing solution, sequentially adds 7.5 phosphate buffer 1s of pH 0 mL, 0.05%
The mL of bromothymol blue solution 4, after 3 min of complexing, adds 10 mL chloroforms to be extracted twice every time, divides and takes chloroform layer, determines 416
Absorbance at nm wavelength, is control with ephedrine hydrochloride, calculates the content of TA；
3）Appropriate Chinese ephedra need testing solution is taken, HPLC analyses are carried out, ephedrine, d-pseudo-ephedrine, methylephedrine is determined
Three kinds of contents of main component；
4）Macroreticular resin is placed in beaker, 24 h are soaked with 95% ethanol, it is fully expanded, 95 % are added after suction filtration
Ethanol washing macroreticular resin to cleaning solution adds appropriate distilled water without white opacity phenomenon, then cleans ethanol with distilled water, and vacuum is taken out
It is standby after filter；
5）Loading, wash-out, wherein loading liquid pH are 5-6, and sample concentration is 0.1-0.15 g mL-1, loading flow velocity is
0.3—0.7 mL•min-1, eluant, eluent volume is 7.5-8 BV, and eluant, eluent is 95% ethanol, and elution flow rate is 0.8-1.2
mL•min-1。
A kind of technique of described macroporous resin purification Herba Ephedrae total alkaloids, it is characterised in that step 3）Middle chromatographic condition
For：Chromatographic column Agilent Zorbax SB-C18（250 mm × 4.6 mm, 5 μm）, 0.1% phosphoric acid：Acetonitrile=95：5, detect ripple
A length of 210 nm；30 DEG C of column temperature；The mL min of flow velocity 1-1, the μ L of sample size 20.
A kind of technique of described macroporous resin purification Herba Ephedrae total alkaloids, it is characterised in that step 4）Described in macropore
Resin is HPD100 type macroreticular resins.
A kind of technique of described macroporous resin purification Herba Ephedrae total alkaloids, it is characterised in that step 5）Middle loading liquid pH
It is 5.3-5.7, sample concentration is 0.11-0.13 g mL-1, loading flow velocity is 0.4-0.6 mL min-1, eluant, eluent volume
It is 7.7-7.9 BV, eluant, eluent is 95% ethanol, and elution flow rate is 0.9-1.1mL min-1。
A kind of technique of described macroporous resin purification Herba Ephedrae total alkaloids, it is characterised in that step 5）Middle loading liquid pH
It is 5.5, sample concentration is 0.12 g mL-1, loading flow velocity is 0.5mL min-1, eluant, eluent volume is 7.8 BV, and eluant, eluent is
95% ethanol, elution flow rate is 1.0mL min-1。
The technique of macroporous resin purification Herba Ephedrae total alkaloids of the invention, there is provided a kind of purifying Herba Ephedrae total alkaloids are most
Good technique, easy to operate, yield is high, is that further galenic pharmacy and pharmacodynamic study are offered reference.
Specific embodiment
Instrument and reagent
The Infinity type high performance liquid chromatographs of Agilent 1260（U.S. Agilent Technologies）；UV-
2800H type ultraviolet-uisible spectrophotometers（You Nike Shanghai Instrument Ltd.）；SHA-A type constant temperature digital display water bath chaders
（The double Shun's experiment Development Co., Ltds in Shanghai）；KQ-250E type numerical control supersonic cleaning machines（Kunshan Ultrasonic Instruments Co., Ltd.）,
RE-52A type Rotary Evaporators（Shanghai Ya Rong bio-instruments factory）；METTLER AL104 type a ten thousandth electronic balances（Mei Te
Le-support benefit instrument company）.
Macroreticular resin HPD100（Cangzhou Bon Adsorption Material Science and Technology Co., Ltd of Hebei province）；Bromothymol blue（Hong Kong
Fisher Science companies）；Acetonitrile is chromatographically pure（TEDIA companies of the U.S.）；Ethanol, phosphoric acid, hydrochloric acid, ammoniacal liquor, chloroform etc. are
It is pure to analyze；Ephedrine hydrochloride reference substance No. 171241-201007, pseudoephedrine hydrochloride reference substance No. 171237-
201208th, mephedrine reference substance No.171247-200301（Nat'l Pharmaceutical ＆ Biological Products Control Institute）, glass chromatography
Post（Internal diameter 1.5cm, Hangzhou Hui Pu chemical industry Instrument Ltd. is customized）.
Medicinal material No. 201304 is purchased from Zhejiang Co., Ltd of Zhen Yuan chain drug stores, by Zhejiang University of Traditional Chinese Medicine's resource and identification
Teaching and research room associate professor Chen Xilin is accredited as ephedra sinicaEphedra sinica StapfHerbaceous stem stem.
Method and result
1）Precision weighs Chinese ephedra fine powder in right amount, 8 times of 0.5% aqueous hydrochloric acid solutions of amount of addition, 60 DEG C, 250W ultrasonic extraction 1h,
Suction filtration, required concentration is concentrated under reduced pressure into Rotary Evaporators；
2）Precision draws appropriate Chinese ephedra need testing solution, sequentially adds 7.5 phosphate buffer 1s of pH 0 mL, 0.05%
The mL of bromothymol blue solution 4, after 3 min of complexing, adds 10 mL chloroforms to be extracted twice every time, divides and takes chloroform layer, determines 416
Absorbance at nm wavelength, is control with ephedrine hydrochloride, calculates the content of TA；
3）Appropriate Chinese ephedra need testing solution is taken, HPLC analyses are carried out, ephedrine, d-pseudo-ephedrine, methylephedrine is determined
Three kinds of contents of main component, chromatographic condition is：Chromatographic column Agilent Zorbax SB-C18（250 mm × 4.6 mm, 5 μ
m）, 0.1% phosphoric acid：Acetonitrile=95：5, Detection wavelength is 210 nm；30 DEG C of column temperature；The mL min of flow velocity 1-1, the μ L of sample size 20；
4）HPD100 type macroreticular resins are placed in beaker, 24 h are soaked with 95% ethanol, it is fully expanded, after suction filtration
Add 95% ethanol to wash macroreticular resin to cleaning solution and add appropriate distilled water without white opacity phenomenon, then ethanol is cleaned with distilled water,
It is standby after vacuum filtration；
5）Loading, wash-out, wherein loading liquid pH are 5.5, and sample concentration is 0.12 g mL-1, loading flow velocity is 0.5mL
min-1, eluant, eluent volume is 7.8 BV, and eluant, eluent is 95% ethanol, and elution flow rate is 1.0mL min-1。
Experiment proves that after, the overall target absorption ration of the Herba Ephedrae total alkaloids of gained is 5.58 mg g-1, comprehensively refer to
Mark is 5.50 mg g than desorption quantity-1, close to predicted value（Integrated value is inhaled according to Herba Ephedrae total alkaloids and three kinds of ratios of main component
Attached amount, assign corresponding weight than desorption quantity entropy assessment after be calculated）.
Eluent Rotary Evaporators are evaporated to obtain Herba Ephedrae total alkaloids purified, the wherein content of TA and three is determined
The content of main component is planted, the yield for calculating gained Herba Ephedrae total alkaloids and its three kinds of main components respectively reaches 30.01% He
28.63%, it is promoted to 2.18 times before purification and 2.25 times.
Macroreticular resin has the features such as physical and chemical stability is high, selectivity is good, regeneration treatment facilitates, and this experiment is used
The Adsorption and desorption characteristics of Herba Ephedrae total alkaloids are had obvious by HPD100 type non-polar macroporous resins compared to other macroreticular resins
Advantage.Equilibrium adsorption capacity is up to 15.49 mg g-1, Static Adsorption rate reaches 89.95 % and static desorption efficiency reaches 85.48 %.
Generally, nonpolar adsorption resin is adapted to adsorb apolar substance from polar solvent, and vice versa.In Chinese ephedra alkaloid into
Polarity is larger, and HPD100 types macroreticular resin has larger adsorbance to it, may have larger specific area 650-700 with it
（m2•g-1）, appropriate average pore size 90-100（）There is substantial connection.
The foregoing is only presently preferred embodiments of the present invention, be not intended to limit the invention, it is all it is of the invention spirit and
Any modification, equivalent and improvement for being made within principle etc., are all contained within protection scope of the present invention.
Claims (3)
1. a kind of technique of macroporous resin purification Herba Ephedrae total alkaloids, it is characterised in that comprise the following steps：
1）Precision weighs Chinese ephedra fine powder in right amount, adds 8 times of 0.5% aqueous hydrochloric acid solutions of amount, and 60 DEG C, 250W ultrasonic extraction 1h take out
Filter, required concentration is concentrated under reduced pressure into Rotary Evaporators；
2）Precision draws appropriate step 1）Obtained solution, sequentially adds the mL of 7.5 phosphate buffer 1s of pH 0,0.05% bromine
The mL of bromothymol blue solution 4, after 3 min of complexing, adds 10 mL chloroforms to be extracted twice every time, divides and takes chloroform layer, determines 416
Absorbance at nm wavelength, is control with ephedrine hydrochloride, calculates the content of TA；
3）Take appropriate step 1）Obtained solution, carries out HPLC analyses, determines ephedrine, d-pseudo-ephedrine, methylephedrine three
Plant the content of main component；Chromatographic condition is：Chromatographic column Agilent Zorbax SB-C18, 250 mm × 4.6 mm, 5 μm,
0.1% phosphoric acid：Acetonitrile=95：5, Detection wavelength is 210 nm；30 DEG C of column temperature；The mL min of flow velocity 1-1, the μ L of sample size 20；
4）Macroreticular resin is placed in beaker, 24 h are soaked with 95 % ethanol, it is fully expanded, 95 % ethanol are added after suction filtration
Washing macroreticular resin to cleaning solution adds appropriate distilled water without white opacity phenomenon, then cleans ethanol with distilled water, after vacuum filtration
Standby, described macroreticular resin is HPD100 type macroreticular resins；
5）Loading, wash-out, wherein loading liquid pH are 5-6, and sample concentration is 0.1-0.15 g mL-1, loading flow velocity is
0.3—0.7 mL•min-1, eluant, eluent volume is 7.5-8 BV, and eluant, eluent is 95% ethanol, and elution flow rate is 0.8-1.2
mL•min-1。
2. a kind of technique of macroporous resin purification Herba Ephedrae total alkaloids as claimed in claim 1, it is characterised in that step 5）In
Loading liquid pH is 5.3-5.7, and sample concentration is 0.11-0.13 g mL-1, loading flow velocity is 0.4-0.6 mL min-1,
Eluant, eluent volume is 7.7-7.9 BV, and eluant, eluent is 95% ethanol, and elution flow rate is 0.9-1 .1mL min-1。
3. a kind of technique of macroporous resin purification Herba Ephedrae total alkaloids as claimed in claim 1, it is characterised in that step 5）In
Loading liquid pH is 5.5, and sample concentration is 0.12 g mL-1, loading flow velocity is 0.5mL min-1, eluant, eluent volume is 7.8
BV, eluant, eluent is 95% ethanol, and elution flow rate is 1.0mL min-1。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410047475.6A CN103800397B (en) | 2014-02-11 | 2014-02-11 | The technique of macroporous resin purification Herba Ephedrae total alkaloids |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410047475.6A CN103800397B (en) | 2014-02-11 | 2014-02-11 | The technique of macroporous resin purification Herba Ephedrae total alkaloids |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103800397A CN103800397A (en) | 2014-05-21 |
| CN103800397B true CN103800397B (en) | 2017-06-06 |
Family
ID=50698092
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410047475.6A Active CN103800397B (en) | 2014-02-11 | 2014-02-11 | The technique of macroporous resin purification Herba Ephedrae total alkaloids |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103800397B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113288922B (en) * | 2021-05-13 | 2022-04-08 | 海南葫芦娃药业集团股份有限公司 | Ephedra extract and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1201029A (en) * | 1997-05-29 | 1998-12-09 | 中国科学院西北高原生物研究所 | Method for preparing Ephedra sinica fine powder by Amberlyst process, and technology for producing its by-product tanning material |
| CN1443749A (en) * | 2002-03-12 | 2003-09-24 | 中国科学院过程工程研究所 | Method for separating and purifying ephedrine by using weak acid cation exchange resin and macroporous resin |
| CN1600777A (en) * | 2003-09-25 | 2005-03-30 | 中国科学院过程工程研究所 | New method for picking-up alkakoid of Chinese ephedra in large scale |
-
2014
- 2014-02-11 CN CN201410047475.6A patent/CN103800397B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1201029A (en) * | 1997-05-29 | 1998-12-09 | 中国科学院西北高原生物研究所 | Method for preparing Ephedra sinica fine powder by Amberlyst process, and technology for producing its by-product tanning material |
| CN1443749A (en) * | 2002-03-12 | 2003-09-24 | 中国科学院过程工程研究所 | Method for separating and purifying ephedrine by using weak acid cation exchange resin and macroporous resin |
| CN1600777A (en) * | 2003-09-25 | 2005-03-30 | 中国科学院过程工程研究所 | New method for picking-up alkakoid of Chinese ephedra in large scale |
Non-Patent Citations (3)
| Title |
|---|
| HPLC法同时测定小儿平喘祛痰颗粒中盐酸麻黄碱、盐酸伪麻黄碱、苦杏仁苷含量;杨德斌等;《中国中药杂志》;20130331;第38卷(第5期);687-690 * |
| 大孔树脂分离提取麻黄碱的研究;任海彧等;《离子交换与吸附》;20021231;第18卷(第2期);97-111 * |
| 麻黄及炙麻黄的工艺研究进展;陆燕萍等;《中外医疗》;20121231(第28期);163-164、166 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103800397A (en) | 2014-05-21 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20130303469A1 (en) | Extraction method for ilex kudingcha c. j. tseng leaves, total saponins and use thereof | |
| CN109406679A (en) | A kind of detection method of high effective liquid chromatography for measuring agalloch eaglewood quality | |
| Zhang et al. | Preparative isolation of cordycepin, N6-(2-hydroxyethyl)-adenosine and adenosine from Cordyceps militaris by macroporous resin and purification by recycling high-speed counter-current chromatography | |
| CN108802255A (en) | The method for measuring ganoderic acid A and Ganoderma lucidum triterpenes components content in compound preparation | |
| CN105384737B (en) | A kind of method of the Hydrolysis kinetics alkaloid from ipecac | |
| CN108670973A (en) | A kind of Chloranthus glaber anti-influenza virus activity extract and preparation method thereof | |
| Kong et al. | Preparative enrichment and separation of astragalosides from Radix Astragali extracts using macroporous resins | |
| CN103800397B (en) | The technique of macroporous resin purification Herba Ephedrae total alkaloids | |
| CN106226426A (en) | A kind of high performance liquid chromatography splits the method for canagliflozin five-membered ring impurity enantiomer | |
| CN104069145B (en) | Application of the eupatorium lindleynun var. trifoliolatum in anti-hepatic-B virus medicine is prepared | |
| CN104447910A (en) | Method for preparation of loganin from traditional Chinese medicine Cornus officinalis | |
| CN101724009A (en) | Process for extracting and purifying triterpenoids from oriental waterplantain rhizome | |
| CN105548378A (en) | Method for separation of Canagliflozin alpha and beta isomers | |
| CN105486767A (en) | Method for separating dapagliflozin and alpha-isomer thereof | |
| CN105175426B (en) | A kind of method of the extraction purification Bergenin from treebine stem | |
| CN109331058B (en) | Traditional Chinese medicine extract with antiviral effect and preparation method and application thereof | |
| CN101747393A (en) | Method for simultaneously preparing chemical references of icariin, epimedin A, epimedin B and epimedin C | |
| CN102920727B (en) | Method for preparing extracts rich in vitexin rhamnoside and vitexin glucoside | |
| CN101401880B (en) | Quality control method for Ditong rhinitis drop and mist | |
| CN103044253B (en) | Extraction separation method of rosmarinic acid in salvia castabea diels f. tomentosa stib | |
| CN102389456A (en) | Method for extracting isodon japonica var.galaucocalyx total diterpenoids or Glaucocalyxin A | |
| CN108794299A (en) | The method of purified solanesol | |
| CN102964404A (en) | Seperation purification preparative technology and quantitative analysis method of cortex albiziae effective monomer | |
| CN102093210A (en) | Purified preparation method of six ginkgoic acid monomers | |
| CN107655986B (en) | Detection method of related substances of vipatavir |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |