CN105457337A - A silica gel loading and sample loading method of a chromatographic column - Google Patents
A silica gel loading and sample loading method of a chromatographic column Download PDFInfo
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- CN105457337A CN105457337A CN201510791845.1A CN201510791845A CN105457337A CN 105457337 A CN105457337 A CN 105457337A CN 201510791845 A CN201510791845 A CN 201510791845A CN 105457337 A CN105457337 A CN 105457337A
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- chromatographic column
- silica gel
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- bottom valve
- sample solution
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/20—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the conditioning of the sorbent material
- B01D15/206—Packing or coating
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D305/00—Heterocyclic compounds containing four-membered rings having one oxygen atom as the only ring hetero atoms
- C07D305/14—Heterocyclic compounds containing four-membered rings having one oxygen atom as the only ring hetero atoms condensed with carbocyclic rings or ring systems
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- Organic Chemistry (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention discloses a silica gel loading and sample loading method of a chromatographic column. The method is characterized by comprising a) a step of preparing a sample solution, b) a step of preparing a solvent-silica gel suspension, c) a step of separation preparation, namely a step of closing a bottom valve of the chromatographic column, adding a solvent into the chromatographic column, slowly adding the solvent-silica gel suspension to the chromatographic column, closing the bottom valve of the chromatographic column after addition of the solvent-silica gel suspension is finished, and allowing the chromatographic column to stand and settle, and d) a step of separation for purification, namely a step of opening the bottom valve of the chromatographic column until a liquid level is rightly below the surface of the silica gel, putting filter paper the diameter of which is slightly smaller than the diameter of the chromatographic column onto the surface of the silica gel, adding the sample solution into the chromatographic column, opening the bottom valve of the chromatographic column, adding the solvent to wash the inner wall of the chromatographic column and a container of the sample solution when the solution lowers to the surface of the silica gel, adding the washing liquid into the chromatographic column, adding an eluant, and performing chromatographic separation for purification. The method is stable in separation quality, standardized in operation and simpler and more convenient in operation.
Description
Technical field
The invention belongs to chromatographic column technical field, specifically, relate to a kind of chromatographic column silica gel and load upper quadrat method.
Background technology
Chromatography is that the object that chemical industry is conventional with pharmaceutical industry is separated, purification process, wherein one of target of the modernization of Chinese medicine is exactly the effective elements of the medicine separation and purification in handle, develop efficient, low toxicity, taking convenience new medicinal preparation, silica gel column chromatography separation is the main method of effective active component separation purifying in plant extracts.
Silica gel column chromatography separation of produced; the filling of carrier of separating silica gel and loading are two very crucial steps; be directly connected to effect and the quality of silica gel column chromatography separation; in current scale silica gel column chromatography separation production process; the filling of carrier of separating silica gel and loading do not have normalized operation; cause disintegrate-quality unstable, have a strong impact on production.
Summary of the invention
For solving above technical problem, the object of the present invention is to provide a kind of chromatographic column silica gel to load upper quadrat method, the steady quality of separation, standardized operation, and operation is more simple and convenient.
The present invention seeks to realize like this:
Upper quadrat method loaded by a kind of chromatographic column silica gel, and its key is to carry out according to the following steps:
A, preparation sample solution: in sample to be separated, add dissolution with solvents obtain sample solution;
B, preparation solvent-silica gel suspended matter: in silica gel, add solvent;
C, separation prepare: close chromatographic column bottom valve, the solvent of 1/3 chromatographic column volume is added in chromatographic column, then in chromatographic column, solvent-silica gel suspended matter is slowly added, slowly open chromatographic column bottom valve simultaneously, keep adding speed equal with discharge rate, maintenance liquid level exceeds silica gel face 14.5 ~ 15.5cm, after dissolving-silica gel suspended matter adds simultaneously, close chromatographic column bottom valve, chromatographic column standing sedimentation;
D, separating-purifying: open chromatographic column bottom valve and just close down to during silica gel face to liquid level, silica gel face is padded the filter paper that a diameter is slightly less than column diameter; Again sample solution is added in chromatographic column, open chromatographic column bottom valve, when solution drops to silica gel face, then add the container of solvent wash chromatographic column inwall and sample solution, after cleaning solution is included into chromatographic column, adds eluent and start chromatography separating-purifying.
Preferably, the solvent in above-mentioned steps a and d adds volume≤4% chromatographic column volume.
Preferably, the solvent volume in above-mentioned steps b and silica gel weight are than being 3.5mL/g.
Beneficial effect:
A kind of chromatographic column silica gel of the present invention loads upper quadrat method, the steady quality of separation, standardized operation, and operation is more simple and convenient.
Detailed description of the invention
The major impurity of content 70% taxol crude product is Cephalomannine and 7-Epitaxol, taxol and 7-Epitaxol are isomers, taxol and Cephalomannine molecular structure closely similar, the chromatography difficulty between them is larger, in this, as example, there is certain representativeness.Below in conjunction with embodiment, the invention will be further described.
Embodiment 1:
<1> takes the taxol crude product of 1000g content 70%, adds 3500ml carrene stirring and dissolving complete, obtains sample solution.
<2> takes 35kg200 ~ 300 order chromatographic silica gel, adds 23L dichloromethane solvent, stirs, and obtains carrene-silica gel suspended matter.
<3> closes the chromatographic column bottom valve of diameter 273mm, high 150cm, add methylene chloride 29L, add carrene-silica gel suspended matter, open chromatographic column bottom valve, controlling opening of valve, keep solvent liquid level about 15cm on silica gel face, carrene-silica gel suspended matter adds complete, close chromatographic column bottom valve, allow silicagel column bed standing sedimentation 12 hours.
<4> opens chromatographic column bottom valve, when liquid level drops to silica gel face, silica gel face is padded the filter paper that a diameter is slightly less than chromatographic column diameter, add sample solution, when sample solution liquid level drops to silica gel face, with 3L washed with dichloromethane column wall and beaker, after cleaning solution enters chromatographic column completely, add eluent dichloromethane-ethyl acetate-isopropanol mixed solution, start chromatography.
The segmentation of <5> chromatography stream part merges, concentrated, dry, obtains taxol principal piece 600g, content of taxol 99.15%.
Finally it should be noted that; foregoing description is only the preferred embodiments of the present invention; those of ordinary skill in the art is under enlightenment of the present invention; under the prerequisite without prejudice to present inventive concept and claim; can make like multiple types and representing, such conversion all falls within protection scope of the present invention.
Claims (3)
1. a upper quadrat method loaded by chromatographic column silica gel, it is characterized in that carrying out according to the following steps:
A, preparation sample solution: in sample to be separated, add dissolution with solvents obtain sample solution;
B, preparation solvent-silica gel suspended matter: in silica gel, add solvent;
C, separation prepare: close chromatographic column bottom valve, the solvent of 1/3 chromatographic column volume is added in chromatographic column, then in chromatographic column, solvent-silica gel suspended matter is slowly added, slowly open chromatographic column bottom valve simultaneously, keep adding speed equal with discharge rate, maintenance liquid level exceeds silica gel face 14.5 ~ 15.5cm, after dissolving-silica gel suspended matter adds simultaneously, close chromatographic column bottom valve, chromatographic column standing sedimentation 12h;
D, separating-purifying: open chromatographic column bottom valve and just close down to during silica gel face to liquid level, silica gel face is padded the filter paper that a diameter is slightly less than column diameter; Again sample solution is added in chromatographic column, open chromatographic column bottom valve, when solution drops to silica gel face, then add the container of solvent wash chromatographic column inwall and sample solution, after cleaning solution is included into chromatographic column, adds eluent and start chromatography separating-purifying.
2. upper quadrat method loaded by chromatographic column silica gel according to claim 1, it is characterized in that: the solvent in described step a and d adds volume≤4% chromatographic column volume.
3. upper quadrat method loaded by chromatographic column silica gel according to claim 2, it is characterized in that: the solvent volume in described step b and silica gel weight are than being 3.5mL/g.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510791845.1A CN105457337A (en) | 2015-11-17 | 2015-11-17 | A silica gel loading and sample loading method of a chromatographic column |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510791845.1A CN105457337A (en) | 2015-11-17 | 2015-11-17 | A silica gel loading and sample loading method of a chromatographic column |
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| CN105457337A true CN105457337A (en) | 2016-04-06 |
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| CN201510791845.1A Pending CN105457337A (en) | 2015-11-17 | 2015-11-17 | A silica gel loading and sample loading method of a chromatographic column |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107519665A (en) * | 2017-07-28 | 2017-12-29 | 上海理工大学 | A kind of method of tunning crude separation in situ extracting fermentation |
| CN111974025A (en) * | 2019-05-24 | 2020-11-24 | 正大天晴药业集团南京顺欣制药有限公司 | Method for filling automatic glue spraying chromatographic column |
| CN112645906A (en) * | 2020-12-29 | 2021-04-13 | 重庆臻源红豆杉发展有限公司 | Method for separating and purifying paclitaxel by high-efficiency chromatography |
| CN116392854A (en) * | 2023-06-07 | 2023-07-07 | 江中药业股份有限公司 | Full-automatic chromatographic device and analysis system |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH046463A (en) * | 1990-04-16 | 1992-01-10 | W R Grace & Co | Carrier for liquid chromatography and liquid chromatographic method using said carrier |
| CN101020671A (en) * | 2006-08-03 | 2007-08-22 | 沈阳天峰生物工程技术有限公司 | Process of separating and purifying taxol efficiently |
| CN102219764A (en) * | 2011-05-12 | 2011-10-19 | 河南省科学院化学研究所有限公司 | Method for separating and purifying paclitaxel industrially |
| CN103193572A (en) * | 2013-04-07 | 2013-07-10 | 山东大学 | Separation method of single enantiomer of pantoprazole compound |
| CN103788032A (en) * | 2012-10-31 | 2014-05-14 | 江苏汉邦科技有限公司 | Preparation method of paclitaxel |
-
2015
- 2015-11-17 CN CN201510791845.1A patent/CN105457337A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH046463A (en) * | 1990-04-16 | 1992-01-10 | W R Grace & Co | Carrier for liquid chromatography and liquid chromatographic method using said carrier |
| CN101020671A (en) * | 2006-08-03 | 2007-08-22 | 沈阳天峰生物工程技术有限公司 | Process of separating and purifying taxol efficiently |
| CN102219764A (en) * | 2011-05-12 | 2011-10-19 | 河南省科学院化学研究所有限公司 | Method for separating and purifying paclitaxel industrially |
| CN103788032A (en) * | 2012-10-31 | 2014-05-14 | 江苏汉邦科技有限公司 | Preparation method of paclitaxel |
| CN103193572A (en) * | 2013-04-07 | 2013-07-10 | 山东大学 | Separation method of single enantiomer of pantoprazole compound |
Non-Patent Citations (1)
| Title |
|---|
| 北京大学化学与分子工程学院有机化学研究所: "《有机化学实验》", 30 June 2015, 北京大学出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107519665A (en) * | 2017-07-28 | 2017-12-29 | 上海理工大学 | A kind of method of tunning crude separation in situ extracting fermentation |
| CN111974025A (en) * | 2019-05-24 | 2020-11-24 | 正大天晴药业集团南京顺欣制药有限公司 | Method for filling automatic glue spraying chromatographic column |
| CN112645906A (en) * | 2020-12-29 | 2021-04-13 | 重庆臻源红豆杉发展有限公司 | Method for separating and purifying paclitaxel by high-efficiency chromatography |
| CN116392854A (en) * | 2023-06-07 | 2023-07-07 | 江中药业股份有限公司 | Full-automatic chromatographic device and analysis system |
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Application publication date: 20160406 |
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