CN105964606B - Extend the cleaning method of using life of chromatographic column - Google Patents
Extend the cleaning method of using life of chromatographic column Download PDFInfo
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- CN105964606B CN105964606B CN201610259783.4A CN201610259783A CN105964606B CN 105964606 B CN105964606 B CN 105964606B CN 201610259783 A CN201610259783 A CN 201610259783A CN 105964606 B CN105964606 B CN 105964606B
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- chromatographic column
- cleaning
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- ultra
- pure water
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- 238000004140 cleaning Methods 0.000 title claims abstract description 86
- 238000000034 method Methods 0.000 title claims abstract description 41
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims abstract description 36
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims abstract description 19
- 229910017604 nitric acid Inorganic materials 0.000 claims abstract description 19
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims abstract description 18
- 239000003960 organic solvent Substances 0.000 claims abstract description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 33
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 29
- 239000012498 ultrapure water Substances 0.000 claims description 29
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 24
- 239000002253 acid Substances 0.000 claims description 4
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 2
- 229910052698 phosphorus Inorganic materials 0.000 claims description 2
- 239000011574 phosphorus Substances 0.000 claims description 2
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 2
- 230000002000 scavenging effect Effects 0.000 description 12
- 238000001514 detection method Methods 0.000 description 10
- 239000007788 liquid Substances 0.000 description 10
- 229920000877 Melamine resin Polymers 0.000 description 6
- JDSHMPZPIAZGSV-UHFFFAOYSA-N melamine Chemical compound NC1=NC(N)=NC(N)=N1 JDSHMPZPIAZGSV-UHFFFAOYSA-N 0.000 description 6
- 239000000463 material Substances 0.000 description 4
- 238000004587 chromatography analysis Methods 0.000 description 3
- 230000003628 erosive effect Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 235000013365 dairy product Nutrition 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 206010019133 Hangover Diseases 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000011010 flushing procedure Methods 0.000 description 1
- 235000015243 ice cream Nutrition 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 235000020185 raw untreated milk Nutrition 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B08—CLEANING
- B08B—CLEANING IN GENERAL; PREVENTION OF FOULING IN GENERAL
- B08B3/00—Cleaning by methods involving the use or presence of liquid or steam
- B08B3/04—Cleaning involving contact with liquid
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B08—CLEANING
- B08B—CLEANING IN GENERAL; PREVENTION OF FOULING IN GENERAL
- B08B3/00—Cleaning by methods involving the use or presence of liquid or steam
- B08B3/04—Cleaning involving contact with liquid
- B08B3/08—Cleaning involving contact with liquid the liquid having chemical or dissolving effect
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B08—CLEANING
- B08B—CLEANING IN GENERAL; PREVENTION OF FOULING IN GENERAL
- B08B9/00—Cleaning hollow articles by methods or apparatus specially adapted thereto
- B08B9/02—Cleaning pipes or tubes or systems of pipes or tubes
- B08B9/027—Cleaning the internal surfaces; Removal of blockages
Landscapes
- Engineering & Computer Science (AREA)
- Mechanical Engineering (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The present invention proposes a kind of cleaning method for extending using life of chromatographic column, and this method includes：Using phosphoric acid, the first cleaning is carried out to the chromatographic column；Using nitric acid, the second cleaning is carried out to the chromatographic column by the described first cleaning；And using organic solvent, third cleaning is carried out to the chromatographic column by the described second cleaning.This method can thoroughly remove the residual sample in chromatographic column and pipeline, significantly extend the service life and utilization rate of chromatographic column.
Description
Technical field
The present invention relates to the cleaning methods for extending using life of chromatographic column.
Background technology
Using high performance liquid chromatograph detect sample in melamine content, since sample kind is more, including cheese,
Ice cream, Yoghourt, liquid milk and its various raw materials, and sample composition is complicated, and the residual component of sample is to liquid chromatogram after detection
The pollution of instrument pipeline and chromatographic column is larger, according to the method for existing cleaning instrument, it is impossible to by chromatographic chromatographic column and pipe
Impurity in road thoroughly cleans up, and often occurs peak hangover or forward position phenomenon so as to cause testing result, chromatographic column is caused to make
With the reduction in service life, and then the frequency for replacing chromatographic column is increased, laboratory must replace a chromatography every one to two months
Column, chromatographic column is expensive, laboratory consumptive material high cost.
Therefore, the cleaning method of high performance liquid chromatograph needs further to develop and improve.
Invention content
The present invention is directed to solve at least some of the technical problems in related technologies.For this purpose, the present invention carries
A kind of method that can thoroughly remove sample carryover in chromatographic column is gone out, the service life of chromatographic column greatly prolongs.
In the first aspect of the present invention, the present invention proposes a kind of cleaning method for extending using life of chromatographic column.According to
The embodiment of the present invention, the method includes：Using phosphoric acid, the first cleaning is carried out to the chromatographic column；Using nitric acid, to passing through
The chromatographic column of first cleaning carries out the second cleaning；And using organic solvent, to the chromatographic column by the described second cleaning
Carry out third cleaning.The cleaning method that the embodiment of the present invention proposes can thoroughly remove the sample carryover in chromatographic column and pipeline,
The service life of chromatographic column significantly extends, and laboratory is since the method using the embodiment of the present invention, averagely one as where inventor
It is only and needs to change 1 root chromatogram column.
According to an embodiment of the invention, the cleaning method of above-mentioned extension using life of chromatographic column can further include as
At least one lower additional technical feature：
According to an embodiment of the invention, first cleaning is carried out at least 1 hour using the phosphoric acid of 20~50 volume %.20
The phosphoric acid of~50 volume % can effectively dissolve the residual sample in chromatographic column, and scavenging period can be adjusted accordingly according to flow velocity,
Scavenging period and flow velocity match the residual sample that can effectively clean in chromatographic column, but scavenging period should at least ensure at 1 hour
More than, to ensure effectively to clean residual sample.
According to an embodiment of the invention, second cleaning is carried out at least 1 hour using the nitric acid of 5~10 volume %.5~
The nitric acid of 10 volume % further can effectively dissolve the residual sample not dissolved in chromatographic column by phosphoric acid, and the shape inside pipeline
Into uniform and smooth oxidation film, prevent nitric acid from playing effective and appropriate protective effect to the excessive erosion of pipeline.Scavenging period
At least at 1 hour or more, it is ensured that the further thorough cleaning to residual sample in chromatographic column.According to an embodiment of the invention,
Scavenging period can be adjusted accordingly according to flow velocity, and scavenging period and flow velocity match and can further thoroughly clean in chromatographic column
Residual sample.
According to an embodiment of the invention, the organic solvent is included selected from least one of methanol and acetonitrile.It is of the invention real
The method for applying example finally cleans chromatographic column using methanol or acetonitrile, and methanol is full of in the chromatographic column and its pipeline after cleaning
Or acetonitrile preserves for use, therefore, organic solvent uses at least one of methanol or acetonitrile, can simplify cleaning step, before process
After the cleaning successively of phosphoric acid and nitric acid described in face, the mild methanol of character of use or acetonitrile can effectively protect chromatographic column prevent
Chromatographic column is by the service life of excessive erosion, further extension chromatographic column.
According to an embodiment of the invention, before first cleaning is carried out, further comprise：Using ultra-pure water to described
Chromatographic column carries out prerinse at least 20 minutes.Ultra-pure water prerinse will can be easier at least 20 minutes in chromatographic column and pipeline in advance
The sample carryover being rinsed rinses out, and then further increases phosphoric acid or nitric acid and the contact surface of residual sample in chromatographic column
Product, so it is more thorough to the cleaning of chromatographic column.
According to an embodiment of the invention, it after first cleaning is carried out and before the described second cleaning, further wraps
It includes：At least 20 minutes are cleaned to the chromatographic column using ultra-pure water, to remove the remaining phosphorus in the chromatographic column
Acid.According to an embodiment of the invention, ultra-pure water cleans the chromatographic column at least 20 minutes, is flowed out from chromatograph pipeline
Water be detected as neutrality, be the instruction rinsed well to residual phosphoric acid in chromatographic column.Using residual in ultrapure water chromatographic column
Phosphoric acid is stayed, is conducive to further effective flushing and dissolving of the follow-up nitric acid to residual sample in chromatographic column.
According to an embodiment of the invention, it after second cleaning is carried out and before third cleaning, further wraps
It includes：At least 20 minutes are cleaned to the chromatographic column using ultra-pure water, to remove the remaining nitre in the chromatographic column
Acid.According to an embodiment of the invention, ultra-pure water cleans the chromatographic column at least 20 minutes, is flowed out from chromatograph pipeline
Water be detected as neutrality, be the instruction rinsed well to residual nitric acid in chromatographic column.Using residual in ultrapure water chromatographic column
Nitric acid is stayed, avoids residual nitric acid during chromatographic column preservation to the excessive erosion of chromatographic column, and then further improve color
Compose the service life of column.
In the second aspect of the present invention, the present invention proposes a kind of cleaning method for extending using life of chromatographic column.According to
The embodiment of the present invention, the method includes：(1) prerinse 20 minutes is carried out to chromatographic column using ultra-pure water, (2) are using 30 bodies
Product % phosphoric acid by the prewashed chromatographic column to carrying out the first cleaning 3 hours, and (3) are using ultra-pure water to passing through described first
The chromatographic column of cleaning carries out cleaning 20 minutes, to remove remaining phosphoric acid in the chromatographic column, and (4) are using 5 volume % nitric acid
The second cleaning 3 hours is carried out to the chromatographic column by ultra-pure water cleaning, (5) are using ultra-pure water to the color by the described second cleaning
It composes column clean again 20 minutes, remaining nitric acid and (6) are super to passing through using methanol in the chromatographic column to remove
The chromatographic column that pure water cleans again is finally cleaned.The cleaning method of the embodiment of the present invention can thoroughly remove chromatographic column and pipe
Residual sample in road significantly extends the service life of chromatographic column.
Description of the drawings
Fig. 1 is the detection peak shape figure of the chromatographic column after the cleaning according to embodiments of the present invention using existing cleaning method；
Fig. 2 is the detection peak shape of the chromatographic column after cleaning method using the present invention cleaning according to embodiments of the present invention
Figure；
Fig. 3 is the chromatography post detection melamine after cleaning method using the present invention cleaning according to embodiments of the present invention
The result figure of content；And
Fig. 4 is the chromatography post detection melamine after cleaning method using the present invention cleaning according to embodiments of the present invention
Another result figure of content.
Specific embodiment
The embodiment of the present invention is described below in detail.It is exemplary below by the embodiment of description, it is intended to for solving
The present invention is released, and is not considered as limiting the invention.
Embodiment 1
In the present embodiment, the step of cleaning chromatographic column, is described in detail：
1) ultra-pure water cleaning step：Pump is closed, woven hose is inserted into ultra-pure water, pump valve is rotated counterclockwise, by chromatographic column
And the gas discharge in pipe-line system, after exhaust, pump valve is tightened, pump is opened, ultra-pure water enters pipeline circulating system pair
Chromatographic column carries out prerinse, and scavenging period is made by oneself according to instrument, and in the present embodiment, the scavenging period that inventor uses is 20 points
Clock.
2) 30% phosphoric acid solution cleaning step：Pump is closed, woven hose is inserted into the phosphoric acid of 30 volume %, is rotated counterclockwise
Gas in chromatographic column and pipe-line system is discharged, after exhaust, tightens pump valve by pump valve, and pump is opened, and 30% phosphoric acid enters
Pipeline circulating system cleans chromatographic column, and scavenging period is made by oneself according to instrument, and in the present embodiment, inventor uses clear
It is 3 hours to wash the time.(remarks：If the small flowrate settable time is 3 hours or more, the big flowrate settable time is 1 hour, but
It is at least set in 1 hour or more).
3) ultra-pure water cleaning step：Pump is closed, woven hose is inserted into ultra-pure water, pump valve is rotated counterclockwise, by chromatographic column
And the gas discharge in pipe-line system, after exhaust, pump valve is tightened, pump is opened, ultra-pure water enters pipeline circulating system pair
Chromatographic column is cleaned, until the pH of the liquid flowed out in pipeline is in neutrality.In the present embodiment, scavenging period is 20 minutes,
The pH of the liquid flowed out in pipeline is in neutrality.
4) 5% salpeter solution cleaning step：Pump is closed, woven hose is inserted into the nitric acid of 5 volume %, rotary pump counterclockwise
Gas in chromatographic column and pipe-line system is discharged, after exhaust, tightens pump valve by valve, and pump is opened, and 5% nitric acid enters pipe
The road circulatory system cleans chromatographic column, and scavenging period is made by oneself according to instrument, in the present embodiment, the cleaning that inventor uses
Time is 3 hours.(remarks：If the small flowrate settable time is 3 hours or more, the big flowrate settable time is 1 hour, but extremely
It is set in less 1 hour or more.)
5) ultra-pure water cleaning step：Pump is closed, woven hose is inserted into ultra-pure water, pump valve is rotated counterclockwise, by chromatographic column
And the gas discharge in pipe-line system, after exhaust, pump valve is tightened, pump is opened, ultra-pure water enters pipeline circulating system pair
Chromatographic column is cleaned, until the pH of the liquid flowed out in pipeline is in neutrality.In the present embodiment, scavenging period is 20 minutes,
The pH of the liquid flowed out in pipeline is in neutrality.
6) organic solvent cleaning cleaning step：Pump to be closed, woven hose is inserted into 100% methanol, rotates pump valve counterclockwise,
Gas in chromatographic column and pipe-line system is discharged, after exhaust, tightens pump valve, pump is opened, 100% methanol enters pipeline
The circulatory system cleans chromatographic column, and 100% methanol is saved backup full of chromatographic column and pipeline.(remarks：It can also be used
100% acetonitrile solution detergent line is finally that 100% acetonitrile is saved backup full of chromatographic column and pipeline).
Embodiment 2
In the present embodiment, the chromatographic column after inventor cleans cleaning method using the present invention is with using existing cleaning
Chromatographic column after method cleaning is detected peaky fuzzy number, as a result as depicted in figs. 1 and 2, wherein, Fig. 1 is shown using existing
Chromatographic column after cleaning method cleaning is using after 2 months, and column effect declines, and trailing phenomenon occurs in standard specimen peak area, and Fig. 2, which is shown, to be adopted
For chromatographic column after being cleaned with the cleaning method of the present invention using after 17 months, detection standard specimen peak area is normal.
Embodiment 3
In the present embodiment, inventor carries out melamine using the chromatographic column after cleaning method using the present invention cleaning
Detection, detection method is according to national standard GB6682-2008《Raw milk and melamine in dairy products detection method》With TQM (T)
72-1042-2《Liquid phase method detects and melamine residual amount job instruction in dairy products, feed, raw material》, testing result is such as
Shown in Fig. 3 and Fig. 4.Testing result shows, the chromatographic column after being cleaned using cleaning method proposed by the present invention, standard items peak type with
And mark-on sample object peak peak type is full symmetric, for symmetrical factor in claimed range, testing result meets national standard.
Embodiment 4
In the present embodiment, the replacement frequency of the chromatographic column after cleaning method of the inventor to using the present invention is united
Meter is found：The WATRS2695/2489 high performance liquid chromatographs in inventor's in March, 2013 laboratory one use method of the invention
Clean once, 2 root chromatogram columns were replaced until in November, 2015, and before this, without cleaning side using the present invention
Method will replace 1 root chromatogram column every the 1-2 months；8 colors have been saved altogether in November, -2015 in March, 2 2013 in inventor laboratory
Column is composed, saves 7840 yuan；Mongolia Ox Taiyuan, Jiaozhuo, the WATRS2695/2489 high performance liquid chromatographs used at the inspection of Tangshan
Consumptive material expense is greatly reduced, and 3000-4000 members can be saved at each inspection.
To sum up, it can analyze and obtain：After cleaning method proposed by the invention, chromatographic column and pipeline inner wall will not be residual
Impurity is stayed, the standard sample collection of illustrative plates peak type of detection is normal, and without trailing phenomenon, chromatographic column column effect is high, significantly extends chromatographic column
Service life and chromatographic column utilization rate, saved the use cost of chromatographic column, improved whole work efficiency.
In the description of this specification, reference term " one embodiment ", " example ", " is specifically shown " some embodiments "
The description of example " or " some examples " etc. means specific features, structure, material or the spy for combining the embodiment or example description
Point is contained at least one embodiment of the present invention or example.In the present specification, schematic expression of the above terms are not
It must be directed to identical embodiment or example.Moreover, particular features, structures, materials, or characteristics described can be in office
It is combined in an appropriate manner in one or more embodiments or example.In addition, without conflicting with each other, the skill of this field
Art personnel can tie the different embodiments or examples described in this specification and the feature of different embodiments or examples
It closes and combines.
Although the embodiments of the present invention has been shown and described above, it is to be understood that above-described embodiment is example
Property, it is impossible to limitation of the present invention is interpreted as, those of ordinary skill in the art within the scope of the invention can be to above-mentioned
Embodiment is changed, changes, replacing and modification.
Claims (7)
1. a kind of cleaning method for extending using life of chromatographic column, which is characterized in that including：
Using phosphoric acid, the first cleaning is carried out to the chromatographic column；
Using nitric acid, the second cleaning is carried out to the chromatographic column by the described first cleaning；And
Using organic solvent, third cleaning is carried out to the chromatographic column by the described second cleaning,
Second cleaning uses percent by volume to be carried out at least 1 hour for 5~10% nitric acid.
2. according to the method described in claim 1, it is characterized in that, first cleaning uses percent by volume as 20~50%
Phosphoric acid carry out at least 1 hour.
3. according to the method described in claim 1, it is characterized in that, the organic solvent includes being selected from methanol and acetonitrile at least
One of.
4. according to the method described in claim 1, it is characterized in that, before first cleaning is carried out, further comprise：
Prerinse at least 20 minutes is carried out to the chromatographic column using ultra-pure water.
5. it according to the method described in claim 1, it is characterized in that, is cleaned after first cleaning is carried out with described second
Before, further comprise：
At least 20 minutes are cleaned to the chromatographic column using ultra-pure water, to remove the remaining phosphorus in the chromatographic column
Acid.
6. it according to the method described in claim 1, it is characterized in that, is cleaned after second cleaning is carried out with the third
Before, further comprise：
At least 20 minutes are cleaned to the chromatographic column using ultra-pure water, to remove the remaining nitre in the chromatographic column
Acid.
7. a kind of cleaning method for extending using life of chromatographic column, which is characterized in that including：
(1) prerinse 20 minutes is carried out to chromatographic column using ultra-pure water,
(2) it is that 30% phosphoric acid cleans 3 hours to carrying out first by the prewashed chromatographic column to use percent by volume,
(3) cleaning 20 minutes is carried out to the chromatographic column by the described first cleaning using ultra-pure water, to remove the chromatographic column
In remaining phosphoric acid,
(4) percent by volume is used to carry out the second cleaning 3 hours to the chromatographic column by ultra-pure water cleaning for 5% nitric acid,
(5) cleaning 20 minutes again are carried out to the chromatographic column by the described second cleaning using ultra-pure water, to remove the color
Compose column in remaining nitric acid and
(6) using methanol to finally being cleaned by the chromatographic column that ultra-pure water cleans again.
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| JP6908125B2 (en) * | 2017-10-05 | 2021-07-21 | 株式会社島津製作所 | Supercritical fluid chromatograph and supercritical fluid chromatography analysis method |
| CN112792034A (en) * | 2020-12-15 | 2021-05-14 | 江苏思派新能源科技有限公司 | Cleaning method of gas chromatographic column for analyzing trace impurities in high-purity carbonate |
| CN113061495B (en) * | 2021-03-11 | 2022-09-13 | 山东省食品药品检验研究院 | A cleaning solution for liquid chromatography online filter element and one-way valve |
| CN114295767B (en) * | 2022-03-10 | 2022-06-03 | 湖南明瑞制药有限公司 | Medicine impurity detection method based on propane fumarate tenofovir disoproxil tablets |
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| US4808233A (en) * | 1987-07-02 | 1989-02-28 | W. R. Grace & Co.-Conn. | Method for high pH washing of silica-based chromatographic sorbents |
| TW422712B (en) * | 1997-04-01 | 2001-02-21 | Wako Pure Chem Ind Ltd | Liquid test components separation method and equipment thereof |
| CN1771080B (en) * | 2003-04-08 | 2010-12-15 | 诺沃挪第克公司 | Method for producing therapeutic peptide or its precursor comprising at least one chromatographic step |
| CN102247900A (en) * | 2011-07-25 | 2011-11-23 | 山东福田药业有限公司 | Process for regenerating calcium type chromatographic separation resin |
| CN202943050U (en) * | 2012-11-06 | 2013-05-22 | 方恩华 | Joint for cleaning capillary tube gas chromatographic column |
| CN102944628A (en) * | 2012-11-28 | 2013-02-27 | 青岛轩汇仪器设备有限公司 | Ion chromatograph with automatic cleaning function |
| CN105424839A (en) * | 2015-12-15 | 2016-03-23 | 袁泳 | Regeneration method of reversed-phase packing column of HPLC (high performance liquid chromatograph) |
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