CN115166119B - High-precision low-pressure gradient method of high-performance liquid chromatograph - Google Patents
High-precision low-pressure gradient method of high-performance liquid chromatograph Download PDFInfo
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- CN115166119B CN115166119B CN202210772961.9A CN202210772961A CN115166119B CN 115166119 B CN115166119 B CN 115166119B CN 202210772961 A CN202210772961 A CN 202210772961A CN 115166119 B CN115166119 B CN 115166119B
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- 239000007788 liquid Substances 0.000 title claims abstract description 104
- 238000000034 method Methods 0.000 title claims abstract description 19
- 238000005213 imbibition Methods 0.000 claims description 8
- 230000008569 process Effects 0.000 claims description 7
- 238000010521 absorption reaction Methods 0.000 claims description 4
- 238000005086 pumping Methods 0.000 claims description 2
- 230000003111 delayed effect Effects 0.000 abstract description 2
- 238000010586 diagram Methods 0.000 description 5
- 230000008859 change Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 230000009471 action Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000013375 chromatographic separation Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
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- 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
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
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- 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
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/28—Control of physical parameters of the fluid carrier
- G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
- G01N2030/342—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient fluid composition fixed during analysis
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Abstract
The invention provides a high-precision low-pressure gradient method of a high-performance liquid chromatograph, which relates to the technical field of liquid chromatographs, wherein a high-pressure pump adopts a serial double-plunger pump, a liquid suction side of a first plunger pump and a liquid suction side of a second plunger pump are respectively provided with an electromagnetic valve and a one-way valve, the electromagnetic valve is used for opening liquid flow entering the direction of the first plunger pump from a liquid suction inlet, and the one-way valve is used for controlling liquid flow entering the direction of the second plunger pump from the first plunger pump; before the first plunger pump is not sucked, the electromagnetic valve is opened, so that the pressure of the first plunger pump is reduced to the atmospheric pressure, and the fact that liquid suction is not delayed when the first plunger pump starts sucking is realized, so that the difference of mixing ratio precision caused by the opening delay of the one-way valve is avoided, the time of liquid suction in a small proportion is increased through designing a symmetrical liquid suction interval, the liquid suction precision in the small proportion is improved, and the analysis precision of the high-performance liquid chromatograph is further improved.
Description
Technical Field
The invention relates to a high performance liquid chromatograph, in particular to a high-precision low-pressure gradient method of the high performance liquid chromatograph.
Background
The high performance liquid chromatograph is an instrument which uses the chromatographic separation principle, pushes a mobile phase into a system by a high pressure pump, injects a sample solution into the mobile phase by a sample injector, forms separation on a chromatographic column, and performs analysis of each component on a detector. The high pressure pump commonly used in the industry is a series double plunger pump driven by a cam driven by a motor. According to the designed cam curve, the circumferential rotation of the cam can be converted into the liquid suction and pushing action of the plunger rod. And the double plunger pumps are matched, so that continuous liquid suction and liquid delivery processes with relatively stable system pressure are formed.
The imbibition process is a process of imbibing different mobile phases in a proportion under low pressure conditions, a manner known as low pressure gradients. The low-pressure gradient mode generally forms mixed mobile phases with different proportions according to the opening and closing time of the gradient valve. The mixing precision of the mixed mobile phase, namely the low-pressure gradient mixing ratio precision, has a direct influence on the final analysis result, and the higher the low-pressure gradient mixing ratio precision is, the higher the accuracy of the analysis result is. Therefore, in the case of component analysis by a high performance liquid chromatograph, it is necessary to improve the precision of the low pressure gradient mixing ratio as much as possible.
However, in the existing high performance liquid chromatograph, the precision of the low pressure gradient mixing ratio is not high due to various reasons, for example, when the prior art adopts a one-way valve and the plunger starts to suck, the one-way valve is not opened before the pressure in the pump is reduced to normal pressure, and the mobile phase cannot be sucked into the pump, so that the theoretical suction time and the time suction time are different, which can cause poor mixing precision of the mixed mobile phase. As another example, when mixing is performed in a small proportion (for example, below 5%), the gradient valve response time is insufficient, and the accuracy of mixing in a small proportion is deteriorated. In addition, since the flow phases are of various types, the opening time of the check valve has different values due to different volume elastic coefficients, and the start time of the check valve is different due to the change of other factors such as different system pressures, different temperatures and the like.
Disclosure of Invention
In view of the above, the present invention provides a high-precision low-pressure gradient method for a high-performance liquid chromatograph, so as to improve the precision of the low-pressure gradient mixing ratio, and further improve the analysis precision of the high-performance liquid chromatograph.
For this purpose, the invention provides the following technical scheme:
The high-precision low-pressure gradient method of the high-performance liquid chromatograph is characterized in that a high-pressure pump in the high-performance liquid chromatograph adopts a serial double-plunger pump, a liquid suction side of a first plunger pump and a liquid suction side of a second plunger pump are respectively provided with an electromagnetic valve and a one-way valve, the electromagnetic valve is used for opening liquid flow entering the direction of the first plunger pump from a liquid suction inlet, and the one-way valve is used for controlling liquid flow entering the direction of the second plunger pump from the first plunger pump;
Before the first plunger pump is not sucked, the electromagnetic valve is opened, so that the pressure of the first plunger pump is reduced to the atmospheric pressure.
Further, the electromagnetic valve is a 2-way electromagnetic valve.
Further, the small-scale liquid suction section is designed to be symmetrical front to back.
Further, when the corresponding gradient ratio is changed, the time for sucking the liquid in a small ratio is increased.
Further, a small-scale imbibition interval pair strain gradient model with front-to-back symmetry.
Further, the imbibition process of the first plunger pump is as follows:
When the pumping of the first plunger is completed, the first plunger rod is stopped, the electromagnetic valve and the gradient valve are opened, the system pressure is reduced to the atmospheric pressure, and the one-way valve is automatically closed;
The first plunger rod uniformly accelerates the liquid suction, sucks a small proportion of mobile phase, and increases the valve opening time of a small proportion mixing ratio;
The first plunger rod sucks liquid at a uniform speed, so that the liquid sucking speed is increased;
the first plunger rod uniformly decelerates and absorbs liquid, absorbs a small proportion of mobile phase, and increases the valve opening time of a small proportion mixing ratio;
And (3) completing liquid suction, closing the electromagnetic valve and the gradient valve at the same time, and waiting for liquid delivery.
The invention has the advantages and positive effects that:
1. according to the scheme, the one-way valve is changed into the 2-way electromagnetic valve, so that the pressure can be relieved in advance, the pressure is not influenced by various factors such as pressure, temperature and flow, the liquid suction can be realized without delay, and the liquid suction precision is improved.
2. According to the invention, on the basis of realizing delay-free liquid suction, the liquid suction section is set to be a symmetrical liquid suction section, so that the liquid suction precision of a small proportion (for example, lower than 5%) can be improved, and the separation performance and consistency of the hue chromatograph can be further improved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings may be obtained according to the drawings without inventive effort to a person skilled in the art.
FIG. 1 is a schematic view of the position of a high pressure pump in a liquid chromatograph according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of a tandem double plunger pump according to an embodiment of the present invention;
FIG. 3 is a control block diagram of a high pressure pump in an embodiment of the invention;
FIG. 4 is a schematic diagram of a cam timing section in an embodiment of the present invention;
FIG. 5 is a schematic diagram of a cam imbibition interval curve in an embodiment of the invention;
fig. 6 is a diagram illustrating a gradient change in an embodiment of the present invention.
Detailed Description
In order that those skilled in the art will better understand the present invention, a technical solution in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in which it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the present invention without making any inventive effort, shall fall within the scope of the present invention.
It should be noted that the terms "first," "second," and the like in the description and the claims of the present invention and the above figures are used for distinguishing between similar objects and not necessarily for describing a particular sequential or chronological order. It is to be understood that the data so used may be interchanged where appropriate such that the embodiments of the invention described herein may be implemented in sequences other than those illustrated or otherwise described herein. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, method, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed but may include other steps or elements not expressly listed or inherent to such process, method, article, or apparatus.
As shown in fig. 1, the high performance liquid chromatograph includes: the device comprises a high-pressure pump, a sample injector, a column temperature box and a detector; wherein: the high-pressure pump adopts a series double plunger pump, the motor drives the cam shaft to rotate through the belt, 2 cams are arranged on the cam shaft, and plunger rods of the 2 plunger pumps are respectively driven to push and pull. As shown in fig. 2, a driving motor 1 rotates, and a driving belt 2 drives a cam shaft 3 to rotate; two cams (a first cam 4 and a second cam 5) on the cam shaft synchronously rotate to respectively drive two plunger rods (a first plunger rod 6 and a second plunger rod 7) to reciprocate; the plunger rod is drawn in and pushed in the plunger pump, and liquid is sucked in from the liquid suction port and discharged from the liquid discharge port 14 under the control of the gradient valve 13, thereby achieving the liquid suction and pushing effects. An electromagnetic valve 10 and a check valve 11 are provided on the liquid suction side of the first plunger pump 8 and the liquid suction side of the second plunger pump 9, respectively, the electromagnetic valve 10 being used to open the flow of liquid from the liquid suction port into the direction of the first plunger pump 8, and the check valve 11 being only capable of flowing from the first plunger pump 8 into the direction of the second plunger pump 9. The liquid outlet of the second plunger pump 9 is connected with a pressure sensor 12 which can monitor the pressure in the pipeline at the rear end.
In the existing high performance liquid chromatograph, a one-way valve is arranged on the liquid suction side of the first plunger pump 8, and when the first plunger pump 8 sucks liquid, the pressure of the first plunger pump 8 is the system pressure and has a pressure difference with the atmospheric pressure (0.1 MPa), and the one-way valve cannot be opened immediately, so that poor mixing ratio precision of mobile phases is caused.
In contrast, in the embodiment of the present invention, the one-way valve provided on the liquid suction side of the first plunger pump 8 is replaced with a 2-way electromagnetic valve, and before the first plunger pump 8 is not sucked, the 2-way electromagnetic valve is opened to reduce the pressure of the first plunger pump 8 to the atmospheric pressure. When the first plunger pump 8 starts to suck, liquid suction is not delayed, the liquid suction is not influenced by various factors such as pressure, temperature and flow, and the mixing ratio precision difference caused by the opening delay of the one-way valve is avoided.
When the high performance liquid chromatograph is controlled, as shown in fig. 3, the control system drives the motor to rotate, drives the transmission mechanisms such as a cam shaft in the high pressure pump mechanism to pump the solution out of the liquid storage bottle, flows into the rear end through the pressure sensor, collects the pressure of the system, feeds the pressure back to the control system, and then controls the motor to adjust the rotating speed at a proper time.
In order to further improve the precision of the low-pressure gradient mixing ratio of the high-performance liquid chromatograph, in the embodiment of the invention, on the basis of changing the one-way valve arranged on the liquid suction side of the first plunger pump 8 into a 2-electricity valve, a liquid suction section which is symmetrical front and back is designed, and when the corresponding gradient proportion is changed, the time of a small proportion is increased, so that the precision of the small proportion is improved.
As shown in fig. 4, according to the design of the cam speed regulation curve, the motor theoretically driving the cam shaft rotates at a constant speed, and the first plunger pump 8 and the second plunger pump 9 alternately absorb liquid and push liquid, so that the liquid can be continuously pushed to the rear end at a constant speed. As shown in fig. 5:
Stage ①: when the first plunger pump 8 finishes feeding liquid, the first plunger rod 6 is stopped, the electromagnetic valve 10 and the gradient valve 13 are opened, and the system pressure is reduced to the atmospheric pressure. The check valve 11 is automatically closed, and deviation of mixing ratio precision is avoided.
Stage ②: the plunger rod starts to aspirate.
Stage ③: and (3) completing liquid suction, closing the electromagnetic valve and the gradient valve at the same time, and waiting for liquid delivery.
On the other hand, for a small proportion of mixed mobile phase, the liquid suction precision is inaccurate because the liquid feeding time is too short and the valve closing time is too large. In the embodiment of the invention, the liquid absorption precision of the small proportion is improved by increasing the liquid absorption time of the small proportion (the increased time is calculated according to the designed liquid absorption interval). In addition, the small-scale imbibition interval can be designed to be symmetrical back and forth, and the meaning of the variable gradient is as follows: the a solution was changed from 100% to 0% and the B solution was changed from 0% to 100% in a certain period of time, as shown in fig. 6. Specifically:
As shown in fig. 5, according to the design of the cam imbibition curve, the motor which theoretically drives the cam shaft rotates at a constant speed, and the first plunger pump 8 and the second plunger pump 9 alternately imbibe and push liquid, so that the liquid can be continuously pushed to the rear end at a constant speed.
Stage ①: when the liquid feeding of the first plunger pump 8 is completed, the first plunger rod 6 is stopped, the electromagnetic valve and the gradient valve are opened, the system pressure is reduced to the atmospheric pressure, and the one-way valve is automatically closed.
Stage ②: the first plunger rod 6 evenly accelerates the liquid suction, sucks a small proportion of mobile phase, increases the valve opening time of a small proportion mixing ratio, and improves the precision of the small proportion mixing.
Stage ③: the first plunger rod 6 sucks liquid at a uniform speed, and the liquid sucking speed is increased.
Stage ④: the first plunger rod 6 uniformly decelerates and absorbs liquid, absorbs a small proportion of mobile phase, increases the valve opening time of a small proportion mixing ratio, and improves the precision of the small proportion mixing.
Stage ⑤: and (3) completing liquid suction, closing the electromagnetic valve and the gradient valve at the same time, and waiting for liquid delivery.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.
Claims (4)
1. The high-precision low-pressure gradient method of the high-performance liquid chromatograph is characterized in that a high-pressure pump in the high-performance liquid chromatograph adopts a serial double-plunger pump, a liquid suction side of a first plunger pump and a liquid suction side of a second plunger pump are respectively provided with an electromagnetic valve and a one-way valve, the electromagnetic valve is used for opening liquid flow entering the direction of the first plunger pump from a liquid suction inlet, and the one-way valve is used for controlling liquid flow entering the direction of the second plunger pump from the first plunger pump;
before the first plunger pump is not sucked, the electromagnetic valve is opened, so that the pressure of the first plunger pump is reduced to the atmospheric pressure; the electromagnetic valve is a 2-phase electromagnetic valve;
The imbibition process of the first plunger pump is as follows:
When the pumping of the first plunger is completed, the first plunger rod is stopped, the electromagnetic valve and the gradient valve are opened, the system pressure is reduced to the atmospheric pressure, and the one-way valve is automatically closed;
The first plunger rod uniformly accelerates the liquid suction, sucks a small proportion of mobile phase, and increases the valve opening time of a small proportion mixing ratio;
The first plunger rod sucks liquid at a uniform speed, so that the liquid sucking speed is increased;
the first plunger rod uniformly decelerates and absorbs liquid, absorbs a small proportion of mobile phase, and increases the valve opening time of a small proportion mixing ratio;
And (3) completing liquid suction, closing the electromagnetic valve and the gradient valve at the same time, and waiting for liquid delivery.
2. The high-precision low-pressure gradient method of a high-performance liquid chromatograph according to claim 1, wherein the small-scale liquid absorption section is designed to be symmetrical front to back.
3. The method according to claim 1, wherein the time for sucking the liquid in a small proportion is increased when the corresponding gradient proportion is changed.
4. The high-precision low-pressure gradient method of a high-performance liquid chromatograph according to claim 2, wherein the strain gradient model is composed of a front-back symmetrical small-scale imbibition interval pair.
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| CN115166119B true CN115166119B (en) | 2024-06-14 |
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| CN119438471B (en) * | 2025-01-08 | 2025-05-02 | 科诺美(北京)科技有限公司 | Multi-cycle distribution method of liquid phase low-pressure gradient pump and computer equipment |
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