CN211086205U - Multidimensional online solid phase extraction liquid chromatography device - Google Patents
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- CN211086205U CN211086205U CN201921170982.3U CN201921170982U CN211086205U CN 211086205 U CN211086205 U CN 211086205U CN 201921170982 U CN201921170982 U CN 201921170982U CN 211086205 U CN211086205 U CN 211086205U
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Abstract
The utility model relates to the field of analysis chemical sample pretreatment and liquid chromatography separation and analysis systems, in particular to a multidimensional online solid phase extraction liquid chromatography device and a multidimensional online high performance liquid chromatography analysis device, wherein the analysis device comprises a large-volume liquid sample direct sample introduction assembly, an online purification, extraction and enrichment solid phase extraction assembly and a high performance liquid chromatography separation and analysis assembly; the high performance liquid chromatography separation and analysis component is connected with a first online extraction column. The utility model is suitable for a big volume of complicated sample system like blood sample, urine sample etc. advances appearance, separation and detection. Different online extraction columns and chromatographic analysis column combinations are selected according to different analyzed samples, and the system is flexible to construct. The method can be used together with a mass spectrometer to further improve the resolution ratio, and has very wide application prospect in the separation and analysis of complex systems.
Description
Technical Field
The utility model relates to an analytical chemistry sample pretreatment and liquid chromatography separation analytic system field, concretely relates to multidimensional online solid phase extraction liquid chromatography device is applicable to on-line purification, extraction, enrichment, desorption, separation and the detection of trace organic compound in liquid samples such as medicine, medical treatment, environment, food.
Background
With the development of chromatographic separation technology, the search and separation of trace compounds in complex sample systems has become a hot research field. Multidimensional liquid chromatography is the separation technique developed based on this research hotspot. Compared with the traditional one-dimensional liquid chromatography analysis method, the multi-dimensional liquid chromatography technology effectively improves the separation degree and the peak type of the separation of the complex sample components by improving the peak capacity, and improves the sensitivity. The two-dimensional liquid chromatography system is most common at present, and the method comprises two modes of full two-dimensional and central cutting. The full two-dimensional liquid chromatography refers to that all components in a sample system are subjected to two times of chromatographic separation in different modes, and the central cutting two-dimensional liquid chromatography refers to that a part of components are collected after the first-dimensional separation and enter the second dimension for separation.
The composition of the compounds in actual samples is very complex and the properties vary greatly. For different first dimension fractions, the second dimension separation often does not yield good separation using isocratic or same gradient conditions. Although the resolution of the method can be improved to a certain extent by using a gradient mode with gradual change in the second dimension, the gradient mode is difficult to construct and is less feasible for a two-dimensional liquid chromatography system with better orthogonality. Also, for analysis of complex samples, pretreatment is typically required, such as off-line or on-line separation and enrichment of target analytes in the sample using various extraction materials, followed by injection into a chromatographic system for analysis, most commonly solid phase extraction. However, the on-line combination of solid-phase extraction and high performance liquid chromatography has three limitations: firstly, the offline solid-phase extraction mode needs to manually concentrate and then sample, and the time for loading and processing the sample by the solid-phase extraction column is long, so that the whole analysis time is prolonged; secondly, the online solid phase extraction mode can only carry out large-volume extraction (such as water sampling) on a target object in a cleaner sample, and cannot carry out large-volume extraction (such as blood sampling) on a complex sample; the existing solid-phase extraction online coupling system generally uses a six-way switching valve to connect a single solid-phase extraction column, the two states of sample loading extraction and desorption separation are switched by the six-way switching valve, the online solid-phase extraction column also needs activation, impurity removal and balance time, the whole analysis time is very long, and the analysis flux is greatly reduced.
SUMMERY OF THE UTILITY MODEL
The utility model discloses aim at overcoming the unable online purification through liquid chromatography super high efficiency of complicated sample, on-line enrichment, the shortcoming of online separation and high sensitive analysis, the shortcoming that the peak broadens after the appearance of bulky strong solvent sample advances, and compensate the special demand of extract around the online extraction post, designed one and can realize complicated liquid sample through online automatic sample introduction of online extraction post and chromatographic analysis post combined material, purify, extract, enrichment, desorption, then make the target can be at the online analytic system of super high performance liquid chromatography separation analysis.
The utility model discloses a solve the technical scheme that its technical problem adopted and be:
A multidimensional online solid phase extraction liquid chromatography device comprises a large-volume liquid sample direct sampling component, an online purification, extraction and enrichment solid phase extraction component and a high performance liquid chromatography separation analysis component;
The high performance liquid chromatography separation and analysis component is connected with a first online extraction column.
Furthermore, the direct large-volume sample injection assembly for the liquid sample comprises a first high-pressure chromatographic pump for pushing an extraction liquid, an automatic sample injector for loading a sample, a first high-pressure six-way switching valve for switching a flow path, and a sample ring for realizing large-volume sample injection.
Further, the first high-pressure six-way switching valve is communicated with a first interface and a second interface, and the first interface is connected with a first high-pressure chromatographic pump pipeline; the second interface is connected with the pipeline of the on-line purification, extraction and enrichment solid phase extraction component; and a pipeline between a third interface and a sixth interface of the first high-pressure six-way switching valve is connected with a sample ring, a fifth interface of the first high-pressure six-way switching valve is connected with an automatic sample injector pipeline, and a fourth interface of the first high-pressure six-way switching valve is connected with a pipeline to discharge waste liquid.
Furthermore, a gradient proportional valve and a solvent selection valve are arranged in the first high-pressure chromatographic pump.
Further, the online purification, extraction and enrichment extraction assembly comprises a second high-pressure chromatographic pump for pushing desorption extraction liquid, a second high-pressure six-way switching valve for switching flow paths, a second online extraction column and a third online extraction column for realizing primary purification, enrichment and capture.
Further, the second high-pressure six-way switching valve is communicated with a sixth interface through a first interface, and a fourth interface of the second high-pressure six-way switching valve is respectively connected with the second interface of the first high-pressure six-way switching valve and a pipeline of a second high-pressure chromatographic pump through a three-way pipe; a second online extraction column is connected between a fifth interface and a sixth interface of the second high-pressure six-way switching valve through a pipeline, a third online extraction column is connected between a second interface and a third interface of the second high-pressure six-way switching valve through a pipeline, and a first interface of the second high-pressure six-way switching valve is connected with a high performance liquid chromatography separation and analysis assembly through a pipeline.
Furthermore, the high performance liquid chromatography separation and analysis assembly comprises a first online extraction column, a third high-pressure chromatography pump for pushing elution analysis liquid, a third high-pressure six-way switching valve and a fourth high-pressure six-way switching valve for switching flow paths, a first online chromatography column and a second online chromatography column for realizing separation and analysis of a target object, and a detector for chromatographic detection of the target object.
Further, a gradient proportioning valve is arranged in the third high-pressure chromatographic pump.
Furthermore, the third high-pressure six-way switching valve and the fourth high-pressure six-way switching valve are both communicated with a first interface and a second interface, the second interface of the third high-pressure six-way switching valve is connected with a third high-pressure chromatographic pump pipeline, a first online extraction column is connected between the first interface and the fourth interface of the third high-pressure six-way switching valve through a pipeline, the sixth interface of the third high-pressure six-way switching valve is connected with the first interface pipeline of the second high-pressure six-way switching valve, a fifth interface of the third high-pressure six-way switching valve discharges waste liquid through a connecting pipeline, the third interface of the third high-pressure six-way switching valve is connected with the second interface pipeline of the fourth high-pressure six-way switching valve, a first online chromatographic column is connected between the first interface and the sixth interface of the fourth high-pressure six-way switching valve, and a second online chromatographic column is connected between the third interface and the fourth interface of the fourth high-pressure six-way switching valve, and a fifth interface of the fourth high-pressure six-way switching valve is connected with a detector pipeline.
The utility model has the advantages that:
1. The flow path is switched and various solvents are conveyed by setting the flow rate and the valve state of each chromatographic pump in different time periods (points), the purification, the extraction, the enrichment, the desorption, the separation and the detection are integrated, the whole process is controlled by adopting mechanical drive and software, the automation degree is high, and the precision and the accuracy of the method can be effectively improved; fractions obtained by chromatographic separation are collected by adopting an online extraction column, and an extraction liquid is introduced before the compounds enter a chromatographic analysis column so as to improve the enrichment and capture efficiency.
2. The method allows a large-volume complex liquid sample to be injected into the on-line extraction column, realizes full sample introduction of a target object, has no peak broadening phenomenon, improves the sensitivity of the method and reduces the detection limit.
3. The effluent of the chromatographic column can directly enter a mass spectrum detector without impurity removal and salt removal.
4. Having a bypass desorption flow path allows for optimization of desorption conditions for the target and the extraction material.
5. Different on-line extraction columns and different chromatographic columns can be selected according to the properties of the target, so that the applicability and the application range of the system are widened.
6. The device and the system are not limited to the better application, and can be widely applied to online purification, extraction, enrichment, desorption, separation, matrix effect evaluation, post-column derivation, detection and analysis and the like of target substances in complex samples of environment, food, medicine and the like.
Drawings
FIG. 1 is a schematic view of the flow path communication of the multi-dimensional on-line high performance liquid chromatography system provided by the present invention. (J1: autosampler, T1: detector, P1: first high-pressure chromatography pump, P2: second high-pressure chromatography pump, P3: third high-pressure chromatography pump, A: first high-pressure six-way switching valve, B: second high-pressure six-way switching valve, C: third high-pressure six-way switching valve, D: fourth high-pressure six-way switching valve, H1: sample ring, H2: tee, S1: first online extraction column, S2: second online extraction column, S3: second online extraction column, R1: first online chromatography column, R2: second online chromatography column.)
FIG. 2 is a chromatogram of an analysis of paclitaxel drug concentration in blood using a vitamin on-line solid phase extraction liquid chromatography device;
FIG. 3 is a chromatogram of an analysis of the concentration of paclitaxel drug in blood using a general high performance liquid chromatography.
Detailed Description
In order to make the technical means, creation features, achievement purposes and functions of the present invention easy to understand, the present invention will be further described with reference to the drawings and the specific embodiments.
Example 1
As shown in fig. 1, an online solid phase extraction liquid chromatography apparatus comprises a large volume liquid sample direct sampling assembly, an online purification, extraction and enrichment solid phase extraction assembly and a high performance liquid chromatography separation analysis assembly;
The high performance liquid chromatography separation and analysis component is connected with a first online extraction column S1.
The large-volume liquid sample direct sampling assembly comprises a first high-pressure chromatographic pump P1 for pushing extraction liquid, an automatic sample injector J1 for loading samples, a first high-pressure six-way switching valve A for switching flow paths, and a sample ring H1 for realizing large-volume sampling.
The first high-pressure six-way switching valve A is communicated with a first interface and a second interface, and the first interface is connected with a first high-pressure chromatographic pump P1 through a pipeline; the second interface is connected with the pipeline of the on-line purification, extraction and enrichment solid phase extraction component; and a sample ring H1 is connected between the third interface and the sixth interface of the first high-pressure six-way switching valve A through a pipeline, the fifth interface of the first high-pressure six-way switching valve A is connected with an automatic sample injector J1 through a pipeline, and the fourth interface of the first high-pressure six-way switching valve A is connected with a pipeline to discharge waste liquid.
The first high pressure chromatography pump P1 has a gradient proportioning valve built in.
The online purification, extraction and enrichment extraction assembly comprises a second high-pressure chromatographic pump P2 for pushing desorption extraction liquid, a second high-pressure six-way switching valve B for switching flow paths, a second online extraction column S2 and a third online extraction column S3 for realizing secondary purification, enrichment and capture.
The second high-pressure six-way switching valve B is communicated with a first connector and a sixth connector, and a fourth connector of the second high-pressure six-way switching valve B is respectively connected with the second connector of the first high-pressure six-way switching valve A and a pipeline of a second high-pressure chromatographic pump P2 through a three-way pipe; a second online extraction column S2 is connected between a fifth interface and a sixth interface of the second high-pressure six-way switching valve B through a pipeline, a third online extraction column S3 is connected between a second interface and a third interface of the second high-pressure six-way switching valve B through a pipeline, and a first interface of the second high-pressure six-way switching valve B is connected with a high performance liquid chromatography separation and analysis assembly through a pipeline.
The high performance liquid chromatography separation and analysis assembly comprises a first online extraction column S1, a third high-pressure chromatography pump P3 for pushing elution analysis liquid, a third high-pressure six-way switching valve C and a fourth high-pressure six-way switching valve D for switching flow paths, a first online chromatography column R1 and a second online chromatography column R2 for realizing target separation and analysis, and a detector T1 for target chromatographic detection.
The third high-pressure six-way switching valve C and the fourth high-pressure six-way switching valve D are both communicated with a first port and a second port, the second port of the third high-pressure six-way switching valve C is connected with a third high-pressure chromatographic pump P3 through a pipeline, a first online extraction column S1 is connected between the first port and the fourth port of the third high-pressure six-way switching valve C through a pipeline, the sixth port of the third high-pressure six-way switching valve C is connected with a first port pipeline of the second high-pressure six-way switching valve, the fifth port of the third high-pressure six-way switching valve C discharges waste liquid through a connecting pipeline, the third port of the third high-pressure six-way switching valve C is connected with a second port pipeline of the fourth high-pressure six-way switching valve D, a first online chromatographic column R1 is connected between the first port and the sixth port of the fourth high-pressure six-way switching valve D through a pipeline, and a second online chromatographic column R2 is connected between the third port and the fourth port of the fourth high-pressure six-way switching valve D through a pipeline, and a fifth interface of the fourth high-pressure six-way switching valve D is connected with a detector T1 pipeline.
the first online extraction column used above is an S L CZ1 online extraction column produced by the third medical science and technology ltd, suzhou, and 30 × 4.6mm, the second online extraction column is an S L CZ2 online extraction column produced by the third medical science and technology ltd, suzhou, and 10 × 4.6mm, and the first online chromatographic column is an S L CR1 online chromatographic column produced by the third medical science and technology ltd, suzhou, and 150 × 4.6 mm.
The principle of the utility model is as follows: four high-pressure six-way switching valves are used as a connecting center, and the large-volume liquid sample direct sampling device, the on-line purification, extraction and enrichment solid-phase extraction device and the high performance liquid chromatography separation and analysis device are connected together, so that the on-line purification, extraction, enrichment, desorption, separation and detection of the liquid sample are realized.
The first chromatographic pump is internally provided with a gradient proportional valve for selectively pushing at least two solvents with different components. The sample can be pushed to flow through the first online extraction column, but the target substance enriched in the first online extraction column is not eluted; the cleaning solution is responsible for cleaning the whole extraction pipeline and eluting the target and the matrix remained on the first online extraction column, so that the cross contamination of the next analysis cannot be caused.
an autosampler J1 introduces 150 μ L of a spiked serum sample, a first chromatographic pump P1 pushes 50mM phosphate buffer/acetonitrile (83/17, v/v) loading solution into a second online extraction column S2, then a second high pressure chromatographic pump P2 pushes 30mM phosphate buffer/acetonitrile (50/50, v/v) desorption extract into a first online extraction column S1, the sample is purified and then enriched in a first online extraction column S1, after extraction is complete, a desorption process is entered, a third high pressure six-way switching valve C is switched, after which the first online extraction column S1 is in communication with the first online chromatographic column R1, the first chromatographic pump P1 pushes 50mM phosphate buffer/acetonitrile (50/50, v/v) loading solution through a first online extraction column S1 connected to the second high pressure six-way switching valve B and desorbs the target substance and pushes the target substance into the first online extraction column R4, the v/v-v switching valve C, the elution process is carried out by a third high pressure six-way switching valve C3620, a fourth high pressure chromatographic pump P3610, a high pressure chromatographic column C is connected to separate the target substance from the elution sample, and a target substance is carried out by a high pressure chromatographic column R3670, a high pressure chromatographic column R3610.
Comparative example 1
Replacing the first online extraction column with a two-way joint in the embodiment 1, and when one of the first online extraction columns is replaced with the two-way joint, the connected flow path is changed into a common high performance liquid chromatography system; the measurement was carried out under the same conditions as in example 1, and the sample was directly sent to an on-line analytical column without on-line treatment for separation analysis, and the specific results are shown in FIG. 3.
To sum up, the utility model discloses an effect is compared in comparative example 1 and is more advantageous.
The basic principles, main features and advantages of the present invention have been shown and described above. It will be understood by those skilled in the art that the present invention is not limited to the above embodiments, and that the foregoing embodiments and descriptions are provided only to illustrate the principles of the present invention without departing from the spirit and scope of the present invention. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (9)
1. A multidimensional online solid phase extraction liquid chromatography device is characterized in that: the liquid chromatography device comprises a large-volume liquid sample direct sampling component, an on-line purification, extraction and enrichment solid phase extraction component and a high performance liquid chromatography separation analysis component;
The high performance liquid chromatography separation and analysis component is connected with a first online extraction column.
2. The liquid chromatography apparatus of claim 1, wherein the high-volume liquid sample direct injection assembly comprises a first high-pressure chromatography pump for pushing an extraction liquid, an automatic sample injector for loading a sample, a first high-pressure six-way switching valve for switching flow paths, and a sample loop for realizing high-volume injection.
3. The liquid chromatography apparatus of claim 2, wherein: the first high-pressure six-way switching valve is characterized in that a first interface is communicated with a second interface, and the first interface is connected with a first high-pressure chromatographic pump pipeline; the second interface is connected with the pipeline of the on-line purification, extraction and enrichment solid phase extraction component; and a pipeline between a third interface and a sixth interface of the first high-pressure six-way switching valve is connected with a sample ring, a fifth interface of the first high-pressure six-way switching valve is connected with an automatic sample injector pipeline, and a fourth interface of the first high-pressure six-way switching valve is connected with a pipeline to discharge waste liquid.
4. The liquid chromatography apparatus of claim 2, wherein: the first high-pressure chromatographic pump is internally provided with a gradient proportional valve and a solvent selection valve.
5. The liquid chromatography apparatus of claim 1, wherein: the online purification, extraction and enrichment extraction component comprises a second high-pressure chromatographic pump for pushing desorption extraction liquid, a second high-pressure six-way switching valve for switching flow paths, a second online extraction column and a third online extraction column for realizing primary purification, enrichment and capture.
6. The liquid chromatography apparatus of claim 5, wherein: the second high-pressure six-way switching valve is characterized in that a first connector is communicated with a sixth connector, and a fourth connector of the second high-pressure six-way switching valve is respectively connected with a second connector of the first high-pressure six-way switching valve and a pipeline of a second high-pressure chromatographic pump through a three-way pipe; a second online extraction column is connected between a fifth interface and a sixth interface of the second high-pressure six-way switching valve through a pipeline, a third online extraction column is connected between a second interface and a third interface of the second high-pressure six-way switching valve through a pipeline, and a first interface of the second high-pressure six-way switching valve is connected with a high performance liquid chromatography separation and analysis assembly through a pipeline.
7. The liquid chromatography apparatus of claim 1, wherein: the high performance liquid chromatography separation and analysis assembly comprises a first online extraction column, a third high-pressure chromatography pump for pushing elution analysis liquid, a third high-pressure six-way switching valve and a fourth high-pressure six-way switching valve for switching flow paths, a first online chromatography column and a second online chromatography column for realizing separation and analysis of a target object, and a detector for chromatographic detection of the target object.
8. The liquid chromatography apparatus of claim 7, wherein: the third high-pressure chromatographic pump is internally provided with a gradient proportional valve.
9. The liquid chromatography apparatus of claim 7, wherein: the third high-pressure six-way switching valve and the fourth high-pressure six-way switching valve are both communicated with a first interface and a second interface, the second interface of the third high-pressure six-way switching valve is connected with a third high-pressure chromatographic pump pipeline, a first online extraction column is connected between the first interface and the fourth interface of the third high-pressure six-way switching valve through a pipeline, the sixth interface of the third high-pressure six-way switching valve is connected with a first interface pipeline of the second high-pressure six-way switching valve, a fifth interface of the third high-pressure six-way switching valve discharges waste liquid through a connecting pipeline, the third interface of the third high-pressure six-way switching valve is connected with a second interface pipeline of the fourth high-pressure six-way switching valve, a first online chromatographic column is connected between the first interface and the sixth interface of the fourth high-pressure six-way switching valve through a pipeline, and a second online chromatographic column is connected between the third interface and the fourth interface of the fourth high-pressure six-way switching valve, and a fifth interface of the fourth high-pressure six-way switching valve is connected with a detector pipeline.
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