CN106198831A - A kind of method loading Reversed phase high performance liquid chromatography post - Google Patents
A kind of method loading Reversed phase high performance liquid chromatography post Download PDFInfo
- Publication number
- CN106198831A CN106198831A CN201610824067.6A CN201610824067A CN106198831A CN 106198831 A CN106198831 A CN 106198831A CN 201610824067 A CN201610824067 A CN 201610824067A CN 106198831 A CN106198831 A CN 106198831A
- Authority
- CN
- China
- Prior art keywords
- homogenate
- liquid chromatography
- high performance
- performance liquid
- reversed phase
- 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.)
- Granted
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/89—Inverse chromatography
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The present invention relates to a kind of method loading Reversed phase high performance liquid chromatography post, mainly solve tailing peak and the technical problem of leading peak that existing chromatographic column exists.Packing method of the present invention comprises the following steps: 1) use Pneumatic booster pump to provide power source；2) preparation of filler homogenate: reverse phase filler joined in oxolane and chloroform homogenate, makes filler distribution uniform for after abundant moistening ultrasonic 0.5～2 hour, obtains translucent homogenate；3) connect the homogenate filling cleaned up and void column pipe, column jecket injects ethanol solution, starts air compressor machine, test sealing；4) above-mentioned homogenate is joined in homogenate tank, open booster pump press-in displacement fluid methanol or acetonitrile, maintenance 30～120min after pressure stabilisation；5) rinse chromatographic column 2～5 times of volumes with weight/mass percentage composition 1～5% water and weight/mass percentage composition 99～95% acetonitrile mixed solution, obtain Reversed phase high performance liquid chromatography post.The present invention improves post effect and separating degree, and chromatographic column is packed into power and efficiency has also been brought up to more than 90% by 80% simultaneously.
Description
Technical field
The present invention relates to a kind of method loading high efficiency chromatography post, be specifically related to a kind of side loading Reversed phase high performance liquid chromatography post
Method.
Background technology
Preparing column packing method is the part of most critical in column technology, is also one of principal element determining post effect height.
In the production practices utilizing preparative hplc, the typically equipment replacement to putting into is less, but selects according to post and separation product
Fixing phase, the best dress column method is most important, with the column efficiency height obtained by ensureing, favorable reproducibility, good penetrability, makes point
Reach minimum from purge process cost.Generally according to kind, granule size and the column dimension of selected filler, use different dresses
Column method.General dress column method used uses dry dress method, but dry dress method grain diameter is less than the filler of 20～25 μm, uses
Dry dress method can not obtain high-efficiency column.Because along with reducing of packing material size, intergranular electrostatic interaction and surface can increase, each other
Tend to assemble and " agglomerating ", stick on connecting tube and post jamb, the most also can repel each other because of strong electrostatic interaction, difficult
To load out post bed uniformly and closely.
Summary of the invention
It is an object of the invention to provide a kind of method loading Reversed phase high performance liquid chromatography post, mainly solve existing chromatographic column
The tailing peak existed and the technical problem of leading peak.
Technical scheme is as follows: a kind of method loading Reversed phase high performance liquid chromatography post, comprises the following steps:
1) Pneumatic booster pump is used to provide power source；
2) preparation of filler homogenate: reverse phase filler is joined in oxolane and chloroform homogenate, super after abundant moistening
Sound 0.5～make filler distribution uniform for 2 hours, obtains translucent homogenate；
3) connecting the homogenate filling cleaned up and void column pipe, inject ethanol solution in column jecket, start air compressor machine, test seals
Property；
4) above-mentioned homogenate is joined in homogenate tank, open booster pump press-in displacement fluid methanol or acetonitrile, treat pressure stability
Rear maintenance 30～120min；
5) chromatographic column 2～5 times are rinsed with weight/mass percentage composition 1～5% water and weight/mass percentage composition 99～95% acetonitrile mixed solution
Volume, obtains Reversed phase high performance liquid chromatography post.
The scheme being more highly preferred to is: described step 2) in oxolane and chloroform volume proportion 1:1～1:5；Preferably 1:1
～1:3.
The scheme being more highly preferred to is: displacement fluid stored refrigerated 4-8 hour in described step 4)；Booster pump set pressure as
35-45Mpa。
Beneficial effects of the present invention: the Reversed phase high performance liquid chromatography post loaded by this method can effectively solve tailing peak and
Leading peak, the most significantly improves post effect and separating degree, and chromatographic column is packed into power and efficiency is also brought up to by 80% simultaneously
More than 90%.
Accompanying drawing explanation
Fig. 1 is FUJI filler chromatographic column post effect figure.
Fig. 2 is DASIO filler chromatographic column post effect figure.
Fig. 3 is YMC filler chromatographic column post effect figure.
Fig. 4 is FUJI filler chromatographic column post effect figure.
Wherein: FUJI is Fuji's 120A, 10um filler, DASIO is great Cao 120A, 10um filler, and YMC is 120A, 10um
Filler.
Detailed description of the invention
Reversed-phase high-performance liquid chromatography filling kind is a lot of the most on the market, in order to preferably prove the method suitability.Choosing
Take the filler filling chromatographic column of several different brands different model.
Embodiment 1
Weigh FUJI filler 150g to be positioned in the beaker of 1L, add oxolane and chloroform mixed solution 800mL, its volume ratio
Example is 1:2, stirs ultrasonic to uniform filling distribution, for translucent liquid.
Connecting column jecket and homogenate tank, wherein chromatographic column column jecket specification is 30mm*250mm.Start booster pump to add in column jecket
Enter 100mL dehydrated alcohol, the air-tightness of test junction.
After having tested, set booster pump pressure parameter as 40MPa.Open booster pump, inject even in chromatographic column column jecket
Serosity, pressure slowly begins to ramp up.Solvent flows out from column jecket lower end, and pressure rises to 40MPa.After pressure stabilisation injection replaces
Liquid maintains 120min, and displacement fluid is acetonitrile, uses front stored refrigerated 4 hours.Replacement uses acetonitrile and water mixed solution after terminating again
Rinsing 3 column volumes, its acetonitrile and water quality ratio is 95:5, i.e. can get Reversed phase high performance liquid chromatography post.
By three kinds of substance testing post effects, measurement result asks for an interview Fig. 1.Test condition is shown in Table 1, and test result is shown in Table 2.
Table 1
Table 2
。
Embodiment 2
Weigh DASIO filler 150g to be positioned in the beaker of 1L, add oxolane and chloroform mixed solution 700mL, its volume
Ratio is 1:3, stirs ultrasonic to uniform filling distribution, for translucent liquid.
Connecting column jecket and homogenate tank, wherein chromatographic column column jecket specification is 30mm*250mm.Start booster pump to add in column jecket
Enter 100mL dehydrated alcohol, the air-tightness of test junction.
After having tested, set booster pump pressure parameter as 40MPa.Open booster pump, inject even in chromatographic column column jecket
Serosity, pressure slowly begins to ramp up.Solvent flows out from column jecket lower end, and pressure rises to 40MPa.After pressure stabilisation injection replaces
Liquid maintains 80min, and displacement fluid is methanol, uses front stored refrigerated 6 hours.Replacement is rushed with acetonitrile and water mixed solution after terminating again
Washing 4 column volumes, its acetonitrile and water quality ratio is 97:3, i.e. can get Reversed phase high performance liquid chromatography post.
By three kinds of substance testing post effects, measurement result asks for an interview Fig. 2.Test condition is shown in Table 3, and test result is shown in Table 4.
Table 3
Table 4
。
Embodiment 3
Weigh YMC filler 100g to be positioned in the beaker of 1L, add oxolane and chloroform mixed solution 400mL, its volume ratio
Example is 1:1, stirs ultrasonic to uniform filling distribution, for translucent liquid.
Connecting column jecket and homogenate tank, wherein chromatographic column column jecket specification is 20mm*250mm.Start booster pump to add in column jecket
Enter 100mL dehydrated alcohol, the air-tightness of test junction.
After having tested, set booster pump pressure parameter as 45MPa.Open booster pump, inject even in chromatographic column column jecket
Serosity, pressure slowly begins to ramp up.Solvent flows out from column jecket lower end, and pressure rises to 45MPa.After pressure stabilisation injection replaces
Liquid maintains 100min, and displacement fluid is acetonitrile, uses front stored refrigerated 8 hours.Replacement uses acetonitrile and water mixed solution after terminating again
Rinsing 5 column volumes, its acetonitrile and water quality ratio is 98:2, i.e. can get Reversed phase high performance liquid chromatography post.
By three kinds of substance testing post effects, measurement result asks for an interview annex map 3.Test condition is shown in Table 5, and test result is shown in Table 6.
Table 5
Table 6
。
Embodiment 4
Weigh FUJI filler 150g to be positioned in the beaker of 1L, add oxolane and chloroform mixed solution 700mL, its volume ratio
Example is 1:3, stirs ultrasonic to uniform filling distribution, for translucent liquid.
Connecting column jecket and homogenate tank, wherein chromatographic column column jecket specification is 30mm*250mm.Start booster pump to add in column jecket
Enter 100mL dehydrated alcohol, the air-tightness of test junction.
After having tested, set booster pump pressure parameter as 35MPa.Open booster pump, inject even in chromatographic column column jecket
Serosity, pressure slowly begins to ramp up.Solvent flows out from column jecket lower end, and pressure rises to 35MPa.After pressure stabilisation injection replaces
Liquid maintains 60min, and displacement fluid is methanol, uses front stored refrigerated 5 hours.Replacement is rushed with acetonitrile and water mixed solution after terminating again
Washing 5 column volumes, its acetonitrile and water quality ratio is 98:2, i.e. can get Reversed phase high performance liquid chromatography post.
By three kinds of substance testing post effects, measurement result asks for an interview annex map 4.Test condition is shown in Table 7, and test result is shown in Table 8.
Table 7
Table 8
。
Claims (5)
1. the method loading Reversed phase high performance liquid chromatography post, it is characterised in that comprise the following steps:
1) Pneumatic booster pump is used to provide power source；
2) preparation of filler homogenate: reverse phase filler is joined in oxolane and chloroform homogenate, super after abundant moistening
Sound 0.5～make filler distribution uniform for 2 hours, obtains translucent homogenate；
3) connecting the homogenate filling cleaned up and void column pipe, inject ethanol solution in column jecket, start air compressor machine, test seals
Property；
4) above-mentioned homogenate is joined in homogenate tank, open booster pump press-in displacement fluid methanol or acetonitrile, treat pressure stability
Rear maintenance 30～120min；
5) chromatographic column 2～5 times are rinsed with weight/mass percentage composition 1～5% water and weight/mass percentage composition 99～95% acetonitrile mixed solution
Volume, obtains Reversed phase high performance liquid chromatography post.
A kind of method loading Reversed phase high performance liquid chromatography post the most according to claim 1, it is characterised in that: described oxolane
With chloroform volume proportion 1:1～1:5.
A kind of method loading Reversed phase high performance liquid chromatography post the most according to claim 2, it is characterised in that: described oxolane
Proportioning 1:1～1:3 is imitated with chlorine volume.
A kind of method loading Reversed phase high performance liquid chromatography post the most according to claim 1, it is characterised in that: described booster pump sets
Constant-pressure is 35-45MPa.
A kind of method loading Reversed phase high performance liquid chromatography post the most according to claim 1, it is characterised in that: described displacement fluid is cold
Hide and preserve 4-8 hour.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610824067.6A CN106198831B (en) | 2016-09-13 | 2016-09-13 | A kind of method for loading Reversed phase high performance liquid chromatography post |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610824067.6A CN106198831B (en) | 2016-09-13 | 2016-09-13 | A kind of method for loading Reversed phase high performance liquid chromatography post |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106198831A true CN106198831A (en) | 2016-12-07 |
| CN106198831B CN106198831B (en) | 2017-11-28 |
Family
ID=58067719
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610824067.6A Active CN106198831B (en) | 2016-09-13 | 2016-09-13 | A kind of method for loading Reversed phase high performance liquid chromatography post |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106198831B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN110531012A (en) * | 2018-05-25 | 2019-12-03 | 中科院大连化学物理研究所淮安化工新材料研究中心 | Different phosphorus content C18 liquid phases half prepare chromatography column packing method |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4416783A (en) * | 1981-02-12 | 1983-11-22 | Asahi Kasei Kogyo Kabushiki Kaisha | Liquid chromatography column, process for preparing the same and its use for fractionation |
| US4483374A (en) * | 1982-11-01 | 1984-11-20 | Ethyl Corporation | Chromatographic column |
| CN1532545A (en) * | 2003-03-21 | 2004-09-29 | 中国人民解放军军事医学科学院放射医 | Novel method for producing and packing capillary chromatographic column sieve plate |
| CN101776668A (en) * | 2010-03-13 | 2010-07-14 | 兰州中科安泰分析科技有限责任公司 | Filling method of C18 HPLC (High Performance Liquid Chromatography) column |
| CN201886008U (en) * | 2010-12-03 | 2011-06-29 | 复旦大学 | Homogenizing can for capillary liquid chromatographic-column filling device |
| CN102998401A (en) * | 2011-09-15 | 2013-03-27 | 上海安谱科学仪器有限公司 | Efficient environment-protection filling method for amino liquid chromatography column |
| CN104034830A (en) * | 2014-06-12 | 2014-09-10 | 大连依利特分析仪器有限公司 | Method for filling high-temperature liquid-phase chromatographic column of padding with fine particle size and dedicated device for method |
-
2016
- 2016-09-13 CN CN201610824067.6A patent/CN106198831B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4416783A (en) * | 1981-02-12 | 1983-11-22 | Asahi Kasei Kogyo Kabushiki Kaisha | Liquid chromatography column, process for preparing the same and its use for fractionation |
| US4483374A (en) * | 1982-11-01 | 1984-11-20 | Ethyl Corporation | Chromatographic column |
| CN1532545A (en) * | 2003-03-21 | 2004-09-29 | 中国人民解放军军事医学科学院放射医 | Novel method for producing and packing capillary chromatographic column sieve plate |
| CN101776668A (en) * | 2010-03-13 | 2010-07-14 | 兰州中科安泰分析科技有限责任公司 | Filling method of C18 HPLC (High Performance Liquid Chromatography) column |
| CN201886008U (en) * | 2010-12-03 | 2011-06-29 | 复旦大学 | Homogenizing can for capillary liquid chromatographic-column filling device |
| CN102998401A (en) * | 2011-09-15 | 2013-03-27 | 上海安谱科学仪器有限公司 | Efficient environment-protection filling method for amino liquid chromatography column |
| CN104034830A (en) * | 2014-06-12 | 2014-09-10 | 大连依利特分析仪器有限公司 | Method for filling high-temperature liquid-phase chromatographic column of padding with fine particle size and dedicated device for method |
Non-Patent Citations (1)
| Title |
|---|
| 于世林: "《图解高效液相色谱技术与应用》", 31 May 2009 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN110531012A (en) * | 2018-05-25 | 2019-12-03 | 中科院大连化学物理研究所淮安化工新材料研究中心 | Different phosphorus content C18 liquid phases half prepare chromatography column packing method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106198831B (en) | 2017-11-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104374683B (en) | Core pore compression coefficient testing device and testing method thereof | |
| CN106198831A (en) | A kind of method loading Reversed phase high performance liquid chromatography post | |
| CN205826459U (en) | A kind of device soil infiltration affected for learning gas resistance | |
| CN204649538U (en) | Rock three axle acoustic emission experiment device under a kind of confined pressure, hole press strip part | |
| CN103132971B (en) | Carbon dioxide injection improves the test simulator of coal bed methane recovery rate | |
| CN204841030U (en) | Packing chromatography post preparation facilities | |
| JP2013515258A (en) | Method for dry-packing chromatography columns | |
| CN109946215A (en) | A kind of original position coal body gas absorption amount test simulator | |
| CN101900711A (en) | High performance liquid chromatography method of oxytetracycline residues in soils | |
| CN114965960B (en) | Multi-period injection-production seepage simulation experiment evaluation method for reservoir reconstruction gas storage | |
| CN101526469B (en) | Nondestructive testing device of formaldehyde emission from furniture product and nondestructive testing method thereof | |
| CN208043353U (en) | A kind of dry-type encapsulated property detection device of fuel filter | |
| CN110531012A (en) | Different phosphorus content C18 liquid phases half prepare chromatography column packing method | |
| CN103675169A (en) | Method for filling gel chromatographic columns | |
| CN106442822A (en) | Homogenate tank filled with high performance liquid chromatographic column, filling device and filling method | |
| CN202136832U (en) | Normal pressure column chromatography device under closed environment | |
| CN108152330A (en) | Experimental system and method for measuring influence of osmotic voltage on shale | |
| CN209546783U (en) | A kind of tealeaves fried green equipment | |
| CN207502107U (en) | A kind of construction irrigation device | |
| CN104838885A (en) | High-pressure quick water replenisher for edible mushroom sticks | |
| CN204556182U (en) | A kind of device for detecting sealability of gasoline filter | |
| CN113008941B (en) | Method for testing saturation-resistance increase coefficient of bedding joint development shale | |
| CN108008065A (en) | Ammonia gas detection device and ammonia gas detection method | |
| CN205484229U (en) | A solid -phase extraction post that is arranged in infant food vanillic aldehyde and ethyl vanillin to survey | |
| CN209247774U (en) | Aptitude tests device is perfused in grouting material |
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 |