CN112843789A - Regeneration method of chromatographic packing and chromatographic medium in cannabidiol purification - Google Patents
Regeneration method of chromatographic packing and chromatographic medium in cannabidiol purification Download PDFInfo
- Publication number
- CN112843789A CN112843789A CN202011594151.6A CN202011594151A CN112843789A CN 112843789 A CN112843789 A CN 112843789A CN 202011594151 A CN202011594151 A CN 202011594151A CN 112843789 A CN112843789 A CN 112843789A
- Authority
- CN
- China
- Prior art keywords
- chromatographic
- column
- packing
- cannabidiol
- separation
- 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
Classifications
-
- 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
- B01D15/203—Equilibration or regeneration
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C37/00—Preparation of compounds having hydroxy or O-metal groups bound to a carbon atom of a six-membered aromatic ring
- C07C37/68—Purification; separation; Use of additives, e.g. for stabilisation
- C07C37/70—Purification; separation; Use of additives, e.g. for stabilisation by physical treatment
- C07C37/82—Purification; separation; Use of additives, e.g. for stabilisation by physical treatment by solid-liquid treatment; by chemisorption
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention discloses a regeneration method of chromatographic packing and chromatographic medium in cannabidiol purification, which comprises the following steps: when the separation degree of the filler or the medium is reduced and the separation effect cannot be achieved, the filler is backflushed, 3 column volumes are flushed by alcohol flushing liquid, pollutants are flushed from the column head as far as possible, if the flushing effect in the first step is not obvious, the chromatographic filler is unloaded from the axial compression column, and the integral color change of the column head and the column bed is observed; if the column head is observed to turn black or yellow, the serious pollution part is cut off, the chromatographic packing is homogenized, the chromatographic column is reassembled, 5 column volumes are washed again by hot alcohols, the invention removes the pollutants to the maximum extent according to the principle of similar and compatible solvents, reduces the use of non-relevant solvents as much as possible, reduces the pollution, and improves the service life of the chromatographic packing C18 and C8, chromatographic media PS, PS/DVB and macroporous adsorption resin packing to 1.5 times of the original service life, thereby reducing the separation and purification cost of the CBD of the cannabidiol.
Description
Technical Field
The invention relates to a regeneration method of a chromatographic filler and a chromatographic medium, in particular to a regeneration method of a chromatographic filler and a chromatographic medium in cannabidiol separation, belonging to the field of industrial chromatographic purification and plant extraction.
Background
Cannabidiol (CBD) is an active substance extracted from cannabis plant and has physiological activities of treating epilepsy, insomnia, depression, rheumatoid disease, etc. At present, the conventional cannabidiol purification method at home and abroad is easy to cause serious pollution to the filler or medium, so that the service life of the filler or medium is short.
The silicon dioxide as the chromatographic silica gel matrix is widely applied to the separation of medicines and natural products, the refining purity of CBD extracted from cannabidiol reaches 99 percent, and the high-pressure chromatographic purification of reverse C18 and C8 can not be separated. Chromatographic media such as polymer PS, PS/DVB and macroporous adsorption resin have been applied in the field of plant extraction for many years, and industrial hemp has a good separation effect when THC is removed by less than three thousandths, and the separation effect comprises different particle sizes of 10um, 15um, 20um, 30-50um and more than 100 um; the apertures include 60A, 100A, 120A, 200A, 300A, 500A, 1000A.
In the CBD extraction process, before preparative chromatography or medium-pressure chromatography, a sample contains THC, more components of other substances outside a CBD family are pigments, fatty acids, minerals, terpenes, vegetable proteins and the like, some biological substances are dead adsorbed due to interaction of hydrophobic groups with chromatographic packing and a chromatographic medium during use, the impurities have larger retention values than a target substance, after multiple separations, the impurities adsorbed on the packing or the medium usually accumulate at a column head position, usually, problems of broad peaks, baseline disturbance, tailing and the like are caused, the separation effect of a product is influenced, the packing of the whole column is replaced after the influence of the packing on the separation, and under the background that the price of the packing is high, the method has a large influence on the purification cost of the CBD.
Disclosure of Invention
The invention aims to provide a regeneration method of chromatographic packing and chromatographic medium in cannabidiol separation, which aims to solve the problems that after multiple times of separation, impurities adsorbed on the packing or medium usually accumulate at the position of a column head, which usually causes the problems of broad peak, baseline disturbance, tailing and the like, the separation effect of products is influenced, the packing of the whole column is replaced after the influence of the packing on the separation, and the cost loss of the method is large under the background of high price of the packing.
In order to achieve the purpose, the invention provides the following technical scheme: the regeneration method of the chromatographic packing and the chromatographic medium in the separation of the cannabidiol comprises the following specific steps:
the first step is as follows: when the separation degree of the filler is reduced and the separation effect cannot be achieved, the back flushing filler is firstly used, 3 column volumes are flushed by alcohol flushing liquid, and pollutants are flushed from the column head as clean as possible;
the second step is as follows: if the flushing effect in the first step is not obvious, unloading the chromatographic packing from the axial compression column, and observing the integral color change of the column head and the column bed;
the third step: in the second step, if the column cap is observed to turn black or yellow, the seriously polluted part is cut off;
the fourth step: homogenizing the chromatographic packing, reloading the chromatographic column, and washing 5 column volumes again by hot alcohols;
in a preferred embodiment of the present invention, in the first step, the alcohol rinse solution refers to monohydric alcohol with carbon number not greater than 4, which is miscible with water in any proportion, and includes but is not limited to methanol, ethanol and isopropanol.
In a preferred embodiment of the present invention, in the fourth step, the hot alcohol is one of methanol, ethanol and isopropanol, and the temperature of the hot alcohol is maintained at 50 to 60 ℃.
As a preferred technical scheme of the invention, the chromatographic packing or medium can be one of C18, C8, C4, polymer PS, PS/DV and macroporous adsorption resin which take silica gel as a matrix.
As a preferable technical scheme of the invention, the temperature of hot alcohols is kept at 50-60 ℃, the temperature can be reached by heating the chromatographic column in water bath, and a special heatable solvent tank can be added into the mobile phase solvent.
As a preferred technical scheme of the invention, the temperature of hot alcohols is kept at 50-60 ℃, and the method is realized by adding a heater behind a diaphragm pump of a chromatographic system, and the temperature of the mobile phase can be adjusted at any time by the heater.
Compared with the prior art, the invention has the beneficial effects that:
the invention relates to a regeneration method of chromatographic packing and chromatographic medium in cannabidiol separation, which removes the pollutants to the maximum extent according to the principle of similar compatibility of hydrophobic impurities generated in the extraction process and the purification process of cannabidiol, reduces the use of non-related solvents to the greatest extent and reduces the pollution. The method improves the service life of chromatographic fillers C18, C8 and C4C and chromatographic media PS, PS/DVB and macroporous adsorption resin fillers to 1.5 times of the original service life, thereby reducing the separation and purification cost of cannabidiol CBD.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention provides a technical scheme of a regeneration method of chromatographic packing and chromatographic medium in cannabidiol separation, which comprises the following steps: the regeneration method of chromatographic packing and chromatographic medium in the separation of cannabidiol comprises the following steps:
the first step is as follows: when the separation degree of the filler is reduced and the separation effect cannot be achieved, the back flushing filler is firstly used, 3 column volumes are flushed by alcohol flushing liquid, and pollutants are flushed from the column head as clean as possible;
specifically, the alcohol flushing fluid is monohydric alcohol with the number of carbon atoms not more than 4, which is miscible with water in any proportion, and includes but is not limited to methanol, ethanol and isopropanol;
the second step is as follows: if the flushing effect in the first step is not obvious, unloading the chromatographic packing from the axial compression column, and observing the integral color change of the column head and the column bed;
the third step: in the second step, if the column cap is observed to turn black or yellow, the seriously polluted part is cut off;
the fourth step: homogenizing the chromatographic packing, reloading the chromatographic column, and washing 5 column volumes again by hot alcohols; the hot alcohol is one of methanol, ethanol and isopropanol, and the temperature of the hot alcohol is kept at 50-60 ℃;
wherein, the chromatographic packing or medium can be reverse phase C18, C8 and C4 taking silica gel as a matrix or one of polymer PS, PS/DV and macroporous absorption resin.
Specifically, the temperature of hot alcohols is kept at 50-60 ℃, the temperature can be achieved by heating the chromatographic column in water bath, a special heatable solvent tank can be added to the mobile phase solvent, a heater can be added behind a diaphragm pump of a chromatographic system, and the temperature of the mobile phase can be adjusted at any time through the heater.
In the hemp separation process, ethanol is mostly selected as a mobile phase, a chromatographic column and a mobile phase solvent are required to be heated to 50-60 ℃ for regenerating a filler, the chromatographic column can be heated by a water bath, a special heatable solvent tank can be additionally arranged for the mobile phase solvent, and in the other mode, a heater is additionally arranged behind a diaphragm pump of a chromatographic system, so that the temperature of the mobile phase can be adjusted at any time;
according to the regeneration method of the chromatographic filler and the chromatographic medium in the cannabidiol separation, hydrophobic impurities generated in the cannabidiol extraction process and the purification process are removed to the greatest extent according to the principle of similarity and compatibility, the use of non-relevant solvents is reduced as much as possible, the pollution is reduced, and the service lives of the chromatographic fillers C18 and C8 and the chromatographic media PS, PS/DVB and macroporous adsorption resin filler are prolonged to 1.5 times of the original service lives, so that the separation and purification cost of the cannabidiol CBD is reduced.
Claims (6)
1. The regeneration method of the chromatographic packing and the chromatographic medium in the cannabidiol purification is characterized by comprising the following specific steps:
the first step is as follows: when the separation degree of the filler is reduced and the separation effect cannot be achieved, the filler is backflushed, 3 column volumes are flushed by alcohol flushing liquid, and pollutants are flushed from the column bottom to the column head in a reverse direction as far as possible;
the second step is as follows: if the flushing effect of the column body in the first step is not obvious, unloading the chromatographic packing from the axial compression column, and comparing and observing the overall color change of the column head and the column bed;
the third step: in the second step, if the column cap is observed to turn black or yellow, the seriously polluted part is cut off;
the fourth step: the chromatographic packing is homogenized, the appropriate packing is added and the column is reloaded and washed again for 5 column volumes with hot alcohols.
2. A method of regenerating chromatographic packing and chromatographic media in cannabidiol separations as set forth in claim 1 wherein: in the first step, the alcohol rinse solution is monohydric alcohol with carbon number not greater than 4, which is miscible with water in any proportion, and includes but is not limited to methanol, ethanol and isopropanol.
3. A method of regenerating chromatographic packing and chromatographic media in cannabidiol separations as set forth in claim 1 wherein: in the fourth step, the hot alcohol is one of methanol, ethanol and isopropanol, and the temperature of the hot alcohol is kept at 50-60 ℃.
4. A method of regenerating chromatographic packing and chromatographic media in cannabidiol separations as set forth in claim 1 wherein: the chromatographic packing or medium may be one of silica gel matrix C18, C8, C4, polymer PS, PS/DV and macroporous adsorption resin.
5. A method of regenerating chromatographic packing and chromatographic media in cannabidiol separations as set forth in claim 3 wherein: the temperature of hot alcohols is kept at 50-60 ℃, the temperature can be reached by heating a chromatographic column jacket in water bath, and a special heatable solvent tank can be added into the mobile phase solvent.
6. The method for regenerating chromatographic packing and chromatographic media in cannabidiol separation as claimed in claim 5, wherein: the temperature of hot alcohol is kept at 50-60 ℃, and the method is realized by adding a heater behind a diaphragm pump of the chromatographic system, and the temperature of the mobile phase can be adjusted at any time by the heater.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011594151.6A CN112843789A (en) | 2020-12-29 | 2020-12-29 | Regeneration method of chromatographic packing and chromatographic medium in cannabidiol purification |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011594151.6A CN112843789A (en) | 2020-12-29 | 2020-12-29 | Regeneration method of chromatographic packing and chromatographic medium in cannabidiol purification |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112843789A true CN112843789A (en) | 2021-05-28 |
Family
ID=75998136
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011594151.6A Pending CN112843789A (en) | 2020-12-29 | 2020-12-29 | Regeneration method of chromatographic packing and chromatographic medium in cannabidiol purification |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112843789A (en) |
Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4474663A (en) * | 1982-02-09 | 1984-10-02 | Japan Exlan Company, Limited | Method of regenerating gels for use in liquid chromatography |
| DE10012293A1 (en) * | 2000-03-14 | 2001-10-04 | Dieter Bathen | Process for the chromatographic separation and purification of material mixtures, e.g. solvent compositions comprises using pressure waves during the separation and/or purification process and/or during regeneration of the adsorbent |
| AU2000246542A1 (en) * | 2000-04-21 | 2001-11-07 | Transgenomic, Inc | MIPC column cleaning system and process |
| CN1771080A (en) * | 2003-04-08 | 2006-05-10 | 诺沃挪第克公司 | Regeneration of chromatographic stationary phases |
| DE102006027496B3 (en) * | 2006-06-14 | 2008-01-10 | Boehringer Ingelheim Pharma Gmbh & Co. Kg | Method for optimizing chromatographic purification processes for biomolecules |
| US20080230478A1 (en) * | 2005-05-24 | 2008-09-25 | Ge Healthcare Bio-Sciences Ab | Regeneration Of A Chromatography Matrix |
| US20150093800A1 (en) * | 2013-09-05 | 2015-04-02 | Genentech, Inc. | Method for chromatography reuse |
| US20150328563A1 (en) * | 2012-12-14 | 2015-11-19 | Ge Healthcare Bio-Sciences Ab | Method for Cleaning of Packed Bed Chromatography Columns |
| CN108254478A (en) * | 2018-01-19 | 2018-07-06 | 吉尔生化（上海）有限公司 | A kind of easy refitting regeneration method for preparing chromatographic column |
| CN111960930A (en) * | 2020-08-27 | 2020-11-20 | 哈尔滨工业大学 | Method for separating and purifying cannabidiol from industrial cannabis sativa leaves |
| CN112074333A (en) * | 2018-05-24 | 2020-12-11 | 百特基公司 | Condition conditioning of packed chromatography columns |
-
2020
- 2020-12-29 CN CN202011594151.6A patent/CN112843789A/en active Pending
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4474663A (en) * | 1982-02-09 | 1984-10-02 | Japan Exlan Company, Limited | Method of regenerating gels for use in liquid chromatography |
| DE10012293A1 (en) * | 2000-03-14 | 2001-10-04 | Dieter Bathen | Process for the chromatographic separation and purification of material mixtures, e.g. solvent compositions comprises using pressure waves during the separation and/or purification process and/or during regeneration of the adsorbent |
| AU2000246542A1 (en) * | 2000-04-21 | 2001-11-07 | Transgenomic, Inc | MIPC column cleaning system and process |
| CN1771080A (en) * | 2003-04-08 | 2006-05-10 | 诺沃挪第克公司 | Regeneration of chromatographic stationary phases |
| US20080230478A1 (en) * | 2005-05-24 | 2008-09-25 | Ge Healthcare Bio-Sciences Ab | Regeneration Of A Chromatography Matrix |
| DE102006027496B3 (en) * | 2006-06-14 | 2008-01-10 | Boehringer Ingelheim Pharma Gmbh & Co. Kg | Method for optimizing chromatographic purification processes for biomolecules |
| US20150328563A1 (en) * | 2012-12-14 | 2015-11-19 | Ge Healthcare Bio-Sciences Ab | Method for Cleaning of Packed Bed Chromatography Columns |
| US20150093800A1 (en) * | 2013-09-05 | 2015-04-02 | Genentech, Inc. | Method for chromatography reuse |
| CN108254478A (en) * | 2018-01-19 | 2018-07-06 | 吉尔生化（上海）有限公司 | A kind of easy refitting regeneration method for preparing chromatographic column |
| CN112074333A (en) * | 2018-05-24 | 2020-12-11 | 百特基公司 | Condition conditioning of packed chromatography columns |
| CN111960930A (en) * | 2020-08-27 | 2020-11-20 | 哈尔滨工业大学 | Method for separating and purifying cannabidiol from industrial cannabis sativa leaves |
Non-Patent Citations (12)
| Title |
|---|
| 余盛刚: "反相高效液相色谱固定相再生新方法", 《现代科学仪器》 * |
| 刘翻: "高效液相色谱柱清洗与再生方法", 《分析测试技术与仪器》 * |
| 卞慎之: "高效液相色谱仪维护及常规故障的判断", 《现代仪器》 * |
| 崔莹: "液相色谱柱的使用与维护", 《分析仪器》 * |
| 张春婷等: "反相色谱柱的清洁和再生", 《黑龙江医药》 * |
| 李雪慧: "浅析反相HPLC色谱柱的清洁和再生", 《河南科技》 * |
| 段祖安: "延长液相色谱柱使用寿命的小窍门", 《实验室科学》 * |
| 王春天: "延长液相色谱柱使用寿命的方法", 《甘肃农业》 * |
| 郑伟等: "高效液相色谱柱的选择、使用与维护", 《口腔护理用品工业》 * |
| 陆森林: "高效液相色谱仪色谱柱的维护与保养", 《成都军区医院学报》 * |
| 陈锡龙: "高效液相色谱柱的管理、维护、再生及修复技术", 《贵州畜牧兽医》 * |
| 韩漠: "高效液相色谱法应用中常见问题与处理", 《内蒙古石油化工》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US8236977B2 (en) | Recovery of desired co-products from fermentation stillage streams | |
| US9394505B2 (en) | Recovery of co-products from fermentation stillage streams | |
| US4529551A (en) | Process for separating oleic acid from linoleic acid | |
| Senorans et al. | Analysis of fatty acids in foods by supercritical fluid chromatography | |
| CN109569516B (en) | Mixed adsorbent and application thereof, and polyunsaturated fatty acid treatment method and device | |
| CN111487356B (en) | Method for separating coenzyme Q10 by using supercritical fluid chromatography system | |
| CN111039761B (en) | Method for purifying cannabidiol | |
| CN101279891A (en) | Method for purification of glycerol | |
| AU2006250120A1 (en) | Regeneration of a chromatography matrix | |
| BR112015004186B1 (en) | method for reducing contaminants in the production of a glycol product of hydrogenolysis; and method of making propylene glycol or ethylene glycol | |
| Shin et al. | Improved high‐performance liquid chromatography of vitamin E vitamers on normal‐phase columns | |
| CN101020629A (en) | Process of separating acetylpropionic acid with active carbon | |
| CN1276926C (en) | Tannic acid purifying process | |
| Chen et al. | Synthesis of polyacrylamide/polystyrene interpenetrating polymer networks and the effect of textural properties on adsorption performance of fermentation inhibitors from sugarcane bagasse hydrolysate | |
| Chilla et al. | Automated on-line-solid-phase extraction—high-performance liquid chromatography-diode array detection of phenolic compounds in sherry wine | |
| CN112843789A (en) | Regeneration method of chromatographic packing and chromatographic medium in cannabidiol purification | |
| Aniceto et al. | Preparative chromatography: batch and continuous | |
| Quirino et al. | Towards cleaner downstream processing of biomass waste chemical products by liquid chromatography: A review and recommendations | |
| CN101200516A (en) | Polar macroporous adsorption resin and method for synthesizing the same | |
| CN109265494B (en) | Method for extracting kaempferol glucoside compounds from camellia reticulata | |
| CN110746473B (en) | Purification process for reducing content of lincomycin B component | |
| JP6548087B2 (en) | Selective separation of vitamin E | |
| CN104098470A (en) | Method for synthesizing tricyclodecenyl propionate perfume by adopting loaded solid acid catalyst | |
| CN114539034A (en) | Purification process for extracting resveratrol from grape skin | |
| Xu et al. | One-step simultaneous purification of three water-soluble constituents in crude extracts from Danshen by adsorption chromatography on oligo-β-cyclodextrin substituted agarose gel media |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210528 |
|
| RJ01 | Rejection of invention patent application after publication |