CN101968473A - Dual-ring fed sample purification device and method for gel permeation chromatography purification system - Google Patents
Dual-ring fed sample purification device and method for gel permeation chromatography purification system Download PDFInfo
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- CN101968473A CN101968473A CN2010102850996A CN201010285099A CN101968473A CN 101968473 A CN101968473 A CN 101968473A CN 2010102850996 A CN2010102850996 A CN 2010102850996A CN 201010285099 A CN201010285099 A CN 201010285099A CN 101968473 A CN101968473 A CN 101968473A
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Abstract
The invention discloses a dual-ring fed sample purification device and a dual-ring fed sample purification method for a gel permeation chromatography purification system. In the purification process, a sample is fed again (the second time) at certain specific time when the last (the first time) fed sample purification is not finished so that the substance with short second fed sample keeping time is superposed with the substance with long first fed sample keeping time, but the substance with medium fed sample keeping time every time is not affected; therefore, the purification purpose is fulfilled, the purification efficiency is improved and the solvent is saved. The device and the method increase the separation efficiency of the chromatography system, shorten the purification time and save the consumption of the solvent.
Description
Technical field
The present invention relates to relate to a kind of dicyclo sample introduction purification plant and method that is used for the gel permeation chromatography cleaning system, it specifically is exactly the special use-pattern that is used for the gel permeation chromatography cleaning system of pre-treatment purification, after the single injected sampling, according to the retention time distribution situation of chromatographic effluent, purify for the first time the sample retention time do not finish before sample introduction once more.
Background technology
Gel permeation chromatography (Gel Permeation Chromatography), claim the gel purification chromatogram again, size exclusion chromatograph etc. (Size Exclusion Chromatography) SEC, be called for short GPC, it is a more traditional isolation technics, the pillar of GPC is made up of chemically inert hollow bead, utilizes the principle of size exclusion that sample is separated.
Because micromolecular compound can pass from the hole of filler, and macromolecular compound passes from the filler surrounding space, between micromolecule and the big molecule because the path length difference distance of being flowed through, so big molecule and micromolecule can successively flow out from pillar, play centrifugation, because this isolation technics is not subjected to the influence of the polarity of sample, so for the chromatogram user, be a kind of means that very effectively solve the sample that chromatographic column can not separate, especially for fats, the big molecule chaff interference of protein and pigment, these materials are for chromatographic column, and injection port is abnormally dangerous.And the content in the middle of biogenic sample carried out seeming very necessary once step GPC purification before carrying out sample analysis generally than higher so.
Analyze at general agricultural chemicals/residue of veterinary drug, big molecule chaff interference in the sample substrate is mainly pigment, fat, protein, biogenic chaff interference such as polysaccharide, this class material is for the stratographic analysis of back, be to disturb bigger material, not only can bring high noise, ghost peak, assorted peak, and can pollute the injection port of chromatogram, ion gun, and even whole analytic system, and the life-span of chromatographic column and whole system all had a significant impact, though also have other method to purify, alumina column for example, acidic silica gel post etc., but this class methods majority can be treated analytic sample in various degree absorption is arranged, thereby the recovery of analytical approach is reduced.
At present, shortcoming and defect below gel permeation chromatography exists in purifying the sample use, make it detergent power and be not enough to satisfy the experiment needs, it is less than normal that gel chromatographic columns removes fatty applied sample amount, post is saturated easily, influence separation efficiency, the applied sample amount of the GPC chromatographic column fat that uses as tradition is probably about 0.5-1g, but during agricultural residual and beastly residual in analyzing animal sample and the oil, the content of fat often surpasses 1g in the sample, so traditional solution can reduce the sensitivity of method like this for reducing sampling amount.
Summary of the invention
The purpose of this invention is to provide a kind of dicyclo sample introduction purification plant and method that is used for the gel permeation chromatography cleaning system, promptly according to the principle of gel permeation chromatography, material can be according to the order outflow from gel permeation chromatographic column successively of molecular volume size sequence, big molecule interfering material can take the lead in flowing out from gel permeation chromatographic column, be micromolecular target compound subsequently, so for the method for a GPC, when key collects effluent if being to find out, when stop to collect effluent, target compound is separated, to reach the purpose of purification with interfering material.
Technical scheme of the present invention is as follows:
A kind of dicyclo sample introduction purification plant that is used for the gel permeation chromatography cleaning system, include high-pressure injection pin pump, it is characterized in that: described high-pressure injection pin pump is connected with the 4th valve body and first valve body in turn by pipeline, described first valve body is connected with the loop that is composed in series by chromatographic column, ultraviolet-visible liquid-phase sensor, the 3rd valve body and second valve body by pipeline, also be connected with injection annulus on first valve body respectively, first valve body is connected with the high-pressure liquid phase pump by pipeline; Described the 3rd valve body is connected with the sample introduction needle tubing by pipeline.
A kind of dicyclo sample introduction purification method that is used for the gel permeation chromatography cleaning system, it is characterized in that: it specifically may further comprise the steps: choose high-pressure injection pin pump, high-pressure injection pin pump is connected with the 4th valve body and first valve body in turn by pipeline, pipeline is connected with the loop that is composed in series by chromatographic column, ultraviolet-visible liquid-phase sensor, the 3rd valve body and second valve body on first valve body, also be connected with injection annulus on first valve body, the pipeline that passes through of first valve body is connected with the high-pressure liquid phase pump, and the 3rd valve body is connected with the sample introduction needle tubing by pipeline;
The sample introduction needle tubing is moved on to the top of sample liquid storage bottle to be clean, pump into first sample to be clean by high-pressure injection pin pump, first sample to be clean enters injection annulus by the pipeline circulation, pump into eluting solvent by the high-pressure liquid phase pump, treat that first purifies sample and mixes in injection annulus with eluting solvent, after treating to be full of the potpourri of first sample to be clean and eluting solvent in the injection annulus, adjust the valve location of each valve body, the potpourri circulation of first sample to be clean and eluting solvent enters chromatographic column, thereby first interfering material that purifies in the sample is separated in chromatographic column with target substance;
After a period of time, this moment, first interfering material and target substance that purifies in the sample was not separated fully, pump into second batch of sample to be clean by high-pressure injection pin pump again, second batch of sample to be clean enters injection annulus by the pipeline circulation, pump into eluting solvent by the high-pressure liquid phase pump, treat that second batch of purification sample mixes in injection annulus with eluting solvent, after treating to be full of the potpourri of second batch of sample to be clean and eluting solvent in the injection annulus, adjust the valve location of each valve body, the potpourri circulation of the second batch of sample to be clean and eluting solvent enters chromatographic column, treats that second batch of interfering material in the potpourri that purifies sample and eluting solvent separates in chromatographic column with target substance thereby make; The sample injection time of second batch of sample to be clean should guarantee that wherein the chromatographic peak of target substance can not coincide with the chromatographic peak of target substance in first sample to be clean, when treating that interfering material in first and second batch purification sample and target substance are separated fully, collect respectively the target substance that first and second batch purifies in the sample this moment, target substance in two batches of samples to be clean flows out from chromatographic column fully, and the interfering material that first and second batch purifies in the sample is then discharged by pipeline simultaneously;
At last, adjust the valve location of the 4th valve body, pump into clear water to system pipeline, pipeline and each valve body are cleaned by high-pressure injection pin pump.
The described dicyclo sample introduction purification plant that is used for the gel permeation chromatography cleaning system, it is characterized in that: described first valve body is two 6 logical valves, second valve body is two 4 logical valves, the 3rd valve body is two 4 logical valves, the 4th valve body is two 3 logical valves, described pipeline is stainless-steel tube or polyfluortetraethylene pipe, and diameter is 0.1～5mm.
The described dicyclo sample introduction purification plant that is used for the gel permeation chromatography cleaning system, it is characterized in that: the volume of described injection annulus is 1～200ml.
The described dicyclo sample introduction purification method that is used for the gel permeation chromatography cleaning system is characterized in that: carrying out dicyclo circulation sample introduction must meet the following conditions:
(1), the discharge of interfering material can not have influence on the collection of target substance in first sample to be clean in the second batch of sample to be clean;
(2), the acquisition time of target substance can not clash with the sample injection time of second batch of sample to be clean in the sample to be clean.
(3), the discharge of interfering material can not have influence on the collection of target substance in second batch of sample to be clean in first sample to be clean;
The described dicyclo sample introduction purification method that is used for the gel permeation chromatography cleaning system is characterized in that: described eluting solvent is water, ethanol or or acetonitrile solvent.
Beneficial effect of the present invention:
Than the sample injection method and the sampling device of in the past gel permeation chromatography cleaning system, advantage of the present invention is as follows:
(1), increased the separation efficiency of chromatographic system;
(2), shortened the clarification time, and saved the solvent use amount.
Description of drawings
Fig. 1 is a systematic schematic diagram of the present invention.
Fig. 2 is that the sample introduction gel permeation chromatography goes out peak retention time figure for the first time.
Fig. 3 is that the sample introduction gel permeation chromatography goes out peak retention time figure for the second time.
Fig. 4 does not go out peak retention time figure for using twice sample introduction gel permeation chromatography of dicyclo sample injection method.
Fig. 5 goes out peak retention time figure for using twice sample introduction gel permeation chromatography of dicyclo sample injection method.
Fig. 6 is that two kinds of sample injection method gel permeation chromatographies go out peak retention time comparison diagram.
Fig. 7 is the working state schematic representation of each road valve body.
Embodiment
Referring to Fig. 1, high-pressure injection pin pump 1 is connected with the 4th valve body 2 and first valve body 3 in turn by pipeline, the 3rd valve of the 4th valve body 2 links to each other by pipeline with second valve of first valve body 3, be connected with the loop that is composed in series by chromatographic column 4, ultraviolet-visible liquid-phase sensor 5, the 3rd valve body 6 and second valve body 7 by pipeline between the 3rd, six valves of first valve body 3, the 5th valve that is connected with injection annulus 8, the first valve bodies 3 between first, fourth valve of first valve body 3 is connected with high-pressure liquid phase pump 9 by pipeline; The 3rd valve body 6 is connected with sample introduction needle tubing 10 by pipeline.
A kind of dicyclo sample introduction purification method that is used for the gel permeation chromatography cleaning system, it is characterized in that: it specifically may further comprise the steps: choose high-pressure injection pin pump, high-pressure injection pin pump is connected with the 4th valve body and first valve body in turn by pipeline, pipeline is connected with the loop that is composed in series by chromatographic column, ultraviolet-visible liquid-phase sensor, the 3rd valve body and second valve body on first valve body, also be connected with injection annulus on first valve body, the pipeline that passes through of first valve body is connected with the high-pressure liquid phase pump, and the 3rd valve body is connected with the sample introduction needle tubing by pipeline;
The sample introduction needle tubing is moved on to the top of sample liquid storage bottle to be clean, pump into first sample to be clean by high-pressure injection pin pump, first sample to be clean enters injection annulus by the pipeline circulation, pump into eluting solvent by the high-pressure liquid phase pump, treat that first purifies sample and mixes in injection annulus with eluting solvent, after treating to be full of the potpourri of first sample to be clean and eluting solvent in the injection annulus, adjust the valve location of each valve body, the potpourri circulation of first sample to be clean and eluting solvent enters chromatographic column, thereby first interfering material that purifies in the sample is separated in chromatographic column with target substance;
After a period of time, this moment, first interfering material and target substance that purifies in the sample was not separated fully, pump into second batch of sample to be clean by high-pressure injection pin pump again, second batch of sample to be clean enters injection annulus by the pipeline circulation, pump into eluting solvent by the high-pressure liquid phase pump, treat that second batch of purification sample mixes in injection annulus with eluting solvent, after treating to be full of the potpourri of second batch of sample to be clean and eluting solvent in the injection annulus, adjust the valve location of each valve body, the potpourri circulation of the second batch of sample to be clean and eluting solvent enters chromatographic column, treats that second batch of interfering material in the potpourri that purifies sample and eluting solvent separates in chromatographic column with target substance thereby make; The sample injection time of second batch of sample to be clean should guarantee that wherein the chromatographic peak of target substance can not coincide with the chromatographic peak of target substance in first sample to be clean, when treating that interfering material in first and second batch purification sample and target substance are separated fully, collect respectively the target substance that first and second batch purifies in the sample this moment, target substance in two batches of samples to be clean flows out from chromatographic column fully, and the interfering material that first and second batch purifies in the sample is then discharged by pipeline simultaneously;
At last, adjust the valve location of the 4th valve body, pump into clear water to system pipeline, pipeline and each valve body are cleaned by high-pressure injection pin pump.
Carrying out dicyclo circulation sample introduction must meet the following conditions:
(1), the discharge of interfering material can not have influence on the collection of target substance in first sample to be clean in the second batch of sample to be clean;
(2), the acquisition time of target substance can not clash with the sample injection time of second batch of sample to be clean in the sample to be clean.
(3), the discharge of interfering material can not have influence on the collection of target substance in second batch of sample to be clean in first sample to be clean;
First valve body is two 6 logical valves, and second valve body is two 4 logical valves, and the 3rd valve body is two 4 logical valves, and the 4th valve body is two 3 logical valves, and pipeline is stainless-steel tube or polyfluortetraethylene pipe, and diameter is 0.1～5mm; The volume of injection annulus is 1～200ml; Eluting solvent is water, ethanol or or acetonitrile solvent.
The present invention is further illustrated below in conjunction with embodiment:
Referring to Fig. 2, in the GPC purification applications, if the peak of interested target substance is P2, and the compound of P1 peak and P3 peak representative all is an interfering material, the retention time section of actual collection chromatographic column effluent should be between t2 and the t3 so, and the sample injection time once more of traditional GPC purification method flow process need wait for that the P3 peak flows out back and then sample introduction from chromatographic column, has wasted a large amount of analysis times and organic moving phase like this.
Referring to as Fig. 3, suppose that after analysis is finished carried out once same sample introduction again, establishing secondary sample injection time is t20, three chromatographic peaks are respectively by P21, P22, and P23 represents, so their retention time should with three the peak P1s of first time during sample introduction, P2, P3 is identical.
Referring to Fig. 4, twice sample introduction be drawn on the time shaft represent, t0 represents the time of sample introduction for the first time, and t20 represent the time of the sample introduction second time.
Referring to Fig. 5, if for the first time behind the sample introduction, at t0～t4 certain suitable time in time period sample introduction once more, so the delivery time at P21 peak can with certain overlap of peaks of last sample introduction, if adjust the suitable sample injection time second time, the peak P2 and the P22 that so just can guarantee the target substance that will separate do not overlap with other chromatographic peak, so just can collect the target substance P2 and the P22 of purifying.And the chaff interference mass peak P21 of sample introduction may overlap with the chaff interference mass peak P3 peak of preceding single injected sampling once more, together flows into waste liquid, but this does not influence the result that experiment will obtain, and has improved efficient, has saved solvent.
Referring to Fig. 6, be the comparison diagram of two kinds of sample injection methods, can clearly show the difference of two kinds of sample injection methods.Top figure adopts traditional gel chromatography to purify the chromatogram spectrogram that collection method is carried out sample introduction, following figure adopts dicyclo sample introduction purification method to carry out the chromatogram spectrogram of sample introduction, can learn that on scheming the latter can significantly shorten the clarification time, but be not cost to reduce clean-up effect.
Referring to Fig. 7, the course of work of each road valve body is as follows:
(1), the first injection annulus sample introduction:
The first valve body position A, the second valve body position A, the 3rd valve body position A, the 4th valve body position B, the sample introduction needle tubing moves to sample position, draws sample by high-pressure injection pin pump, first injection annulus is full of sample, then first valve body is turned to the B position, first injection annulus is incorporated the high pressure flow path system into, and the moving phase that pumps of pump is carried sample in first injection annulus and entered chromatographic column and separate like this.
(2), the second injection annulus sample introduction:
The first valve body position B, the second valve body position A, the 3rd valve body position A, the 4th valve body position B, the sample introduction needle tubing moves to sample position, draws sample by high-pressure injection pin pump, second injection annulus is full of sample, first valve body turns to the A position then, and second injection annulus is incorporated the high pressure flow path system into, and the moving phase that pumps of pump is carried sample in second injection annulus and entered chromatographic column and separate like this.
(3), sample collection:
First valve body position A or the B, the second valve body position A, the 3rd valve body position B, the 4th valve body position B, the sample introduction needle tubing moves to assembling position, and the chromatographic column effluent can be collected.
(4), sampling system cleans:
During above sampling system imbibition, the position of the 4th valve body is in position B, if clean the sample introduction pipeline, as long as during the discharge opeing of high-pressure injection pin pump, the position of the 4th valve body is in position A, so just can clean each sample introduction pipeline.
Carry out dicyclo circulation sample introduction and must satisfy following condition:
(1), back one pin effluent can not have influence on last pin and collect component;
(2), the last pin sample introduction collection component time can not conflict with back one pin sample injection time;
(3), last pin effluent can not influence back one pin collection component.
Satisfying under the prerequisite of above-mentioned condition, following the purification efficiency that realizes maximum with the minimum time.
Claims (6)
1. dicyclo sample introduction purification plant that is used for the gel permeation chromatography cleaning system, include high-pressure injection pin pump, it is characterized in that: described high-pressure injection pin pump is connected with the 4th valve body and first valve body in turn by pipeline, described first valve body is connected with the loop that is composed in series by chromatographic column, ultraviolet-visible liquid-phase sensor, the 3rd valve body and second valve body by pipeline, also be connected with injection annulus on first valve body respectively, first valve body is connected with the high-pressure liquid phase pump by pipeline; Described the 3rd valve body is connected with the sample introduction needle tubing by pipeline.
2. dicyclo sample introduction purification method that is used for the gel permeation chromatography cleaning system, it is characterized in that: it specifically may further comprise the steps: choose high-pressure injection pin pump, high-pressure injection pin pump is connected with the 4th valve body and first valve body in turn by pipeline, pipeline is connected with the loop that is composed in series by chromatographic column, ultraviolet-visible liquid-phase sensor, the 3rd valve body and second valve body on first valve body, also be connected with injection annulus on first valve body, the pipeline that passes through of first valve body is connected with the high-pressure liquid phase pump, and the 3rd valve body is connected with the sample introduction needle tubing by pipeline;
The sample introduction needle tubing is moved on to the top of sample liquid storage bottle to be clean, pump into first sample to be clean by high-pressure injection pin pump, first sample to be clean enters injection annulus by the pipeline circulation, pump into eluting solvent by the high-pressure liquid phase pump, treat that first purifies sample and mixes in injection annulus with eluting solvent, after treating to be full of the potpourri of first sample to be clean and eluting solvent in the injection annulus, adjust the valve location of each valve body, the potpourri circulation of first sample to be clean and eluting solvent enters chromatographic column, thereby first interfering material that purifies in the sample is separated in chromatographic column with target substance;
After a period of time, this moment, first interfering material and target substance that purifies in the sample was not separated fully, pump into second batch of sample to be clean by high-pressure injection pin pump again, second batch of sample to be clean enters injection annulus by the pipeline circulation, pump into eluting solvent by the high-pressure liquid phase pump, treat that second batch of purification sample mixes in injection annulus with eluting solvent, after treating to be full of the potpourri of second batch of sample to be clean and eluting solvent in the injection annulus, adjust the valve location of each valve body, the potpourri circulation of the second batch of sample to be clean and eluting solvent enters chromatographic column, treats that second batch of interfering material in the potpourri that purifies sample and eluting solvent separates in chromatographic column with target substance thereby make; The sample injection time of second batch of sample to be clean should guarantee that wherein the chromatographic peak of target substance can not coincide with the chromatographic peak of target substance in first sample to be clean, when treating that interfering material in first and second batch purification sample and target substance are separated fully, collect respectively the target substance that first and second batch purifies in the sample this moment, target substance in two batches of samples to be clean flows out from chromatographic column fully, and the interfering material that first and second batch purifies in the sample is then discharged by pipeline simultaneously;
At last, adjust the valve location of the 4th valve body, pump into clear water to system pipeline, pipeline and each valve body are cleaned by high-pressure injection pin pump.
3. the dicyclo sample introduction purification plant that is used for the gel permeation chromatography cleaning system according to claim 1, it is characterized in that: described first valve body is two 6 logical valves, second valve body is two 4 logical valves, the 3rd valve body is two 4 logical valves, the 4th valve body is two 3 logical valves, described pipeline is stainless-steel tube or polyfluortetraethylene pipe, and diameter is 0.1～5mm.
4. the dicyclo sample introduction purification plant that is used for the gel permeation chromatography cleaning system according to claim 1, it is characterized in that: the volume of described injection annulus is 1～200ml.
5. the dicyclo sample introduction purification method that is used for the gel permeation chromatography cleaning system according to claim 2 is characterized in that: carrying out dicyclo circulation sample introduction must meet the following conditions:
(1), the discharge of interfering material can not have influence on the collection of target substance in first sample to be clean in the second batch of sample to be clean;
(2), the acquisition time of target substance can not clash with the sample injection time of second batch of sample to be clean in the sample to be clean.
(3), the discharge of interfering material can not have influence on the collection of target substance in second batch of sample to be clean in first sample to be clean;
6. the dicyclo sample introduction purification method that is used for the gel permeation chromatography cleaning system according to claim 2, it is characterized in that: described eluting solvent is water, ethanol or acetonitrile solvent.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105606733A (en) * | 2016-01-13 | 2016-05-25 | 中山大学 | External circulation sampling system and external circulation type two-channel online sampling analysis system |
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| JP2006058238A (en) * | 2004-08-23 | 2006-03-02 | Central Res Inst Of Electric Power Ind | Method and apparatus for separating organic chemical component |
| CN101982769A (en) * | 2010-09-07 | 2011-03-02 | 安徽皖仪科技股份有限公司 | Cutting, sub-sampling and purifying device and method for gel chromatography purifying system |
| CN201803993U (en) * | 2010-09-07 | 2011-04-20 | 安徽皖仪科技股份有限公司 | Secondary sample injection cutting purification device for gel chromatography purification system |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2006058238A (en) * | 2004-08-23 | 2006-03-02 | Central Res Inst Of Electric Power Ind | Method and apparatus for separating organic chemical component |
| CN101982769A (en) * | 2010-09-07 | 2011-03-02 | 安徽皖仪科技股份有限公司 | Cutting, sub-sampling and purifying device and method for gel chromatography purifying system |
| CN201803993U (en) * | 2010-09-07 | 2011-04-20 | 安徽皖仪科技股份有限公司 | Secondary sample injection cutting purification device for gel chromatography purification system |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105606733A (en) * | 2016-01-13 | 2016-05-25 | 中山大学 | External circulation sampling system and external circulation type two-channel online sampling analysis system |
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Address after: High tech Zone Wenqu 230088 Hefei Road, Anhui province No. 8 Patentee after: ANHUI WANYI SCIENCE & TECHNOLOGY Co.,Ltd. Address before: Tianda high tech Zone 230088 Hefei Road, Anhui province No. 71 Huayi Science Park building B building Anhui instrument Patentee before: ANHUI WANYI SCIENCE & TECHNOLOGY Co.,Ltd. |