CN211086206U - Three-dimensional online ultra-high performance liquid chromatograph - Google Patents
Three-dimensional online ultra-high performance liquid chromatograph Download PDFInfo
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- CN211086206U CN211086206U CN201921171618.9U CN201921171618U CN211086206U CN 211086206 U CN211086206 U CN 211086206U CN 201921171618 U CN201921171618 U CN 201921171618U CN 211086206 U CN211086206 U CN 211086206U
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- HWXIGFIVGWUZAO-UHFFFAOYSA-N doxofylline Chemical compound C1=2C(=O)N(C)C(=O)N(C)C=2N=CN1CC1OCCO1 HWXIGFIVGWUZAO-UHFFFAOYSA-N 0.000 description 2
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Abstract
The utility model relates to the field of analytical chemistry sample pretreatment and liquid chromatography separation analysis systems, in particular to a three-dimensional online ultra-high performance liquid chromatograph, which 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 analysis assembly; the large-volume liquid sample direct sampling assembly and the high performance liquid chromatography separation analysis assembly are connected with a first online extraction column through pipelines. 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 three-dimensional online super high performance liquid chromatograph 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 three-dimensional online ultra-high performance liquid chromatograph 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 large-volume liquid sample direct sampling assembly and the high performance liquid chromatography separation analysis assembly are connected with a first online extraction column through pipelines.
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 high performance liquid chromatography separation and analysis assembly through a pipeline; 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 secondary 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 high performance liquid chromatography separation and analysis component and a second high-pressure chromatography pump pipeline 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 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.
Furthermore, the third high-pressure six-way switching valve and the fourth high-pressure six-way switching valve are 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, the first 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 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, the fourth interface of the third high-pressure six-way switching valve is connected with the fourth interface of the second high-pressure six-way switching valve and a second high-pressure chromatographic pump pipeline through a three-way pipe fitting, a first online extraction column is connected between a fifth interface of the third high-pressure six-way switching valve and a second interface of the first high-pressure six-way switching valve through a pipeline, and a sixth interface of the third high-pressure six-way switching valve is communicated with a pipeline to discharge waste liquid; a first online chromatographic column is connected between a first interface and a sixth interface of the fourth high-pressure six-way switching valve through a pipeline, a second online chromatographic column is connected between a third interface and a fourth interface of the fourth high-pressure six-way switching valve through a pipeline, and a fifth interface of the fourth high-pressure six-way switching valve is connected with a detector through a pipeline.
Further, a gradient proportional valve is arranged in the third chromatographic pump.
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 extracting agent 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 pipe fitting, S1: first online extraction column, S2: second online extraction column, S3: third online extraction column, R1: first online chromatography column, R2: second online chromatography column.)
FIG. 2 is a chromatogram of analysis of doxofylline drug concentration in blood using a three-dimensional online ultra-high performance liquid chromatograph;
Fig. 3 is a chromatogram for analyzing the concentration of doxofylline drug in blood using 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, a three-dimensional online ultra-high performance liquid chromatograph 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 large-volume liquid sample direct sampling assembly and the high performance liquid chromatography separation and analysis assembly are connected with a first online extraction column S1 through a pipeline.
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 high performance liquid chromatography separation and analysis assembly through a pipeline; and a pipeline between the third interface and the sixth interface of the first high-pressure six-way switching valve A is connected with a sample ring HA, the fifth interface of the first high-pressure six-way switching valve A is connected with an automatic sample injector J1 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 high performance liquid chromatography separation and analysis component and a second high-pressure chromatography pump P2 pipeline through a tee pipe H2; 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 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 J1 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 interface and a second interface, 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, the first interface of the third high-pressure six-way switching valve C is connected with the first interface pipeline of the second high-pressure six-way switching valve B, a third interface of the third high-pressure six-way switching valve C is connected with a second interface pipeline of the fourth high-pressure six-way switching valve D, the fourth interface of the third high-pressure six-way switching valve C is connected with the fourth interface of the second high-pressure six-way switching valve B and a second high-pressure chromatographic pump P2 through a tee pipe H2, a first online extraction column S1 is connected between a fifth interface of the third high-pressure six-way switching valve C and a second interface of the first high-pressure six-way switching valve A through a pipeline, and a sixth interface of the third high-pressure six-way switching valve C is communicated with a pipeline to discharge waste liquid; a first online chromatographic column R1 is connected between a first port and a sixth port of the fourth high-pressure six-way switching valve D through a pipeline, a second online chromatographic column R2 is connected between a third port and a fourth port of the fourth high-pressure six-way switching valve D through a pipeline, and a fifth port of the fourth high-pressure six-way switching valve D is connected with a detector J1 through a pipeline.
the first online extraction column (S1) used above is an S L CZ1 online extraction column produced by Suzhou Adlemi medical science and technology Limited and is 10 × 4.6mm, the second online extraction column (S2) is an S L CZ2 online extraction column produced by Suzhou Adlemi medical science and technology Limited and is 30 × 4.6mm, the third online extraction column (S2) is an S L CZ3 online extraction column produced by Suzhou Adlemi medical science and technology Limited and is 30 × 4.6mm, and the first online chromatography column (R1) is an S L CR1 online chromatography column produced by Suzhou Adlemi medical science and technology Limited and is 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, a large-volume liquid sample direct sampling device, an online purification, extraction and enrichment solid phase extraction device and a high performance liquid chromatography separation liquid chromatograph are connected, and online 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 200 μ L of a spiked serum sample, a first chromatographic pump P1 pushes a sample-carrying liquid of 50mM phosphate buffer/acetonitrile (80/20, v/v) into a first online extraction column S1, then a second high-pressure chromatographic pump P2 pushes a desorption extract of 20mM phosphate buffer/acetonitrile (50/50, v/v) into a second online extraction column S2, the sample is purified and then enriched in a second online extraction column S2, after the extraction is completed, a desorption process is entered, a third high-pressure six-way switching valve C is switched, thereafter the second online extraction column S2 is communicated with the first online chromatographic column R1, the first chromatographic pump P1 pushes a sample-carrying liquid of 50mM phosphate buffer/acetonitrile (50/50, v/v) through a second online extraction column S2 connected to the second online extraction column R8656, the target is desorbed and pushed into the first online chromatographic column R4, the v/v-carrying liquid is pushed through a second online extraction column S2 connected to the second high-pressure six-way switching valve C30, the target elution sample-carrying out a target analysis by a high-pressure chromatography column C3, and a target elution is detected by a fourth high-pressure chromatographic pump 30, a high-pressure chromatographic detector 30, and a target elution detector is carried out a target elution is carried out a target analysis on the target elution detection by a high-pressure switching valve C, a high-pressure chromatography column 30.
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 three-dimensional online ultra-high performance liquid chromatograph is characterized in that: the liquid chromatograph comprises a large-volume liquid sample direct sampling assembly, an on-line purification, extraction and enrichment solid phase extraction assembly and a high performance liquid chromatography separation analysis assembly;
The large-volume liquid sample direct sampling assembly and the high performance liquid chromatography separation analysis assembly are connected with a first online extraction column through pipelines.
2. The three-dimensional online ultra high performance liquid chromatograph of claim 1, wherein the high volume liquid sample direct sampling assembly comprises a first high pressure chromatographic pump for pushing an extraction liquid, an autosampler for loading a sample, a first high pressure six-way switching valve for switching flow paths, and a sample loop for realizing high volume sampling.
3. The three-dimensional online ultra high performance liquid chromatograph 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 high performance liquid chromatography separation and analysis assembly through a pipeline; 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 three-dimensional online ultra high performance liquid chromatograph 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 three-dimensional online ultra high performance liquid chromatograph 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 secondary purification, enrichment and capture.
6. The three-dimensional online ultra high performance liquid chromatograph of claim 5, wherein: the second high-pressure six-way switching valve is characterized in that a first interface is communicated with a sixth interface, and a fourth interface of the second high-pressure six-way switching valve is respectively connected with the high-performance liquid chromatography separation and analysis assembly and a second high-pressure chromatography pump pipeline 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 three-dimensional online ultra high performance liquid chromatograph of claim 1, wherein: the high performance liquid chromatography separation and analysis assembly comprises a third high pressure chromatographic pump used for pushing elution analysis liquid, a third high pressure six-way switching valve and a fourth high pressure six-way switching valve used for switching flow paths, a first online chromatographic column and a second online chromatographic column used for realizing target object separation and analysis, and a detector used for target object chromatographic detection.
8. The three-dimensional online ultra high performance liquid chromatograph of claim 7, wherein: the third high-pressure chromatographic pump is internally provided with a gradient proportional valve.
9. The three-dimensional online ultra high performance liquid chromatograph of claim 7, wherein: the third high-pressure six-way switching valve and the fourth high-pressure six-way switching valve are 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, the first 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 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, the fourth interface of the third high-pressure six-way switching valve is connected with the fourth interface of the second high-pressure six-way switching valve and a second high-pressure chromatographic pump pipeline through a three-way pipe fitting, a first online extraction column is connected between a fifth interface of the third high-pressure six-way switching valve and a second interface of the first high-pressure six-way switching valve through a pipeline, and a sixth interface of the third high-pressure six-way switching valve is communicated with a pipeline to discharge waste liquid; a first online chromatographic column is connected between a first interface and a sixth interface of the fourth high-pressure six-way switching valve through a pipeline, a second online chromatographic column is connected between a third interface and a fourth interface of the fourth high-pressure six-way switching valve through a pipeline, and a fifth interface of the fourth high-pressure six-way switching valve is connected with a detector through a pipeline.
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| CN110455942A (en) * | 2019-07-24 | 2019-11-15 | 苏州艾迪迈医疗科技有限公司 | A kind of online Ultra Performance Liquid Chromatography instrument of three-dimensional |
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| CN110455942A (en) * | 2019-07-24 | 2019-11-15 | 苏州艾迪迈医疗科技有限公司 | A kind of online Ultra Performance Liquid Chromatography instrument of three-dimensional |
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Denomination of utility model: A three-dimensional online ultra-high performance liquid chromatograph Granted publication date: 20200724 Pledgee: Bank of Nanjing Co.,Ltd. Jiangning sub branch Pledgor: Nanjing Aidimai Technology Co.,Ltd. Registration number: Y2024980008518 |