CN103134889B - On-line enrichment-substep focus sample introduction-ultra-high performance liquid chromatography combination system and application - Google Patents
On-line enrichment-substep focus sample introduction-ultra-high performance liquid chromatography combination system and application Download PDFInfo
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Abstract
The invention discloses a substep focus sample introduction-ultra-high performance liquid chromatography combination system and a combination method thereof and an on-line enrichment-substep focus sample introduction-ultra-high performance liquid chromatography combination system and a combination method thereof. The enrichment-substep focus sample introduction-ultra-high performance liquid chromatography combination system comprises on-line enrichment devices, a substep focus sample introduction device and an ultra-high performance liquid chromatography device, wherein every two on-line enrichment devices are connected mutually. The problems that the overloaded volume of the ultra-high performance liquid chromatography is caused by eluant can be solved through a strategy of the substep focus sample introduction. An on-line combination analysis of an enrichment method and the ultra-high performance liquid chromatography can be achieved. Under the premise of obtaining sharpened bands, the on-line enrichment and a rapid separating analysis can be carried out, and analytical sensitivity, speeds and automation degrees can be improved.
Description
Technical field
The invention belongs to analytical chemistry sample pre-treatments field, be specifically related to design and analysis and application that a kind of on-line preconcentration-substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system, be applicable to the online pre-service of trace organic substance in the complex samples such as environment, food, medicine, biology, extraction, desorb, separation and detection.
Background technology
Ultra Performance Liquid Chromatography (UHPLC) velocity of separation is fast, highly sensitive, is a kind of powerful analysis tool.But when being applied in complex sample extreme trace analysis, usually need to adopt efficient pretreatment technology to carry out purifying and enrichment, as Solid-Phase Extraction (SPE) and solid-phase microextraction (SPME).Current beneficiation technologies is combined usually off-line manner with UHPLC, and this is that its packing material size is less, and sampling volume is restricted because UHPLC chromatographic column is shorter and smaller than conventional liquid phase chromatographic column.Often volume is comparatively large, intensity is higher for eluent after SPE or SPME enrichment, and online wash-out sample introduction easily causes UHPLC chromatographic peak broadening and distortion, affects degree of separation and sensitivity.In addition, UHPLC working pressure is higher, and not high voltage bearing fiber material is difficult to and UHPLC on-line coupling.Still lack the on-line preconcentration analysis that effective strategy realizes UHPLC at present, such as, do not occur the report of SPME and UHPLC on-line coupling yet.
Summary of the invention
The present invention is intended to the problem easily causing Ultra Performance Liquid Chromatography peak stretching and distortion for the eluent of on-line preconcentration, design a kind of on-line preconcentration-substep and focus on sample introduction-Ultra Performance Liquid Chromatography combined system, the strategy focused on by substep solves effluent volume overload problem, realize beneficiation technologies and Ultra Performance Liquid Chromatography on-line coupling, relax the restriction of UHPLC to fiber material withstand voltage properties and extraction phase volume, improve degree of separation, analysis efficiency and sensitivity.
Second object of the present invention fills up at present without the blank of SPME and UHPLC on-line coupling, and SPME-UHPLC on-line coupled system combines advantage and the UHPLC feature rapidly and efficiently that SPME operates simple and easy, green low consumption.
3rd object of the present invention focuses on by substep the large volume sample injection UHPLC realizing strong solvent sample to analyze, the spectrum peak distortion avoiding solvent effect to cause, save evaporate to dryness heavy molten or adopt the operation of large water gaging dilute sample, improve automaticity, reduce personal error, improve accuracy, and make large volume sample injection method be applicable to analyze the fluid sample of thing or matrix poorly water-soluble.
The object of the invention is by realize by the following technical solutions:
A kind of substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that comprising:
The substep be connected to each other focuses into sampling device and Ultra Performance Liquid Chromatography device, and described Ultra Performance Liquid Chromatography device comprises chromatographic column, described substep focuses into sampling device and comprises the second high pressure six direction changeover valve and the second quantitative loop, the two ends of the second quantitative loop are connected to the second high pressure six direction changeover valve by stainless steel pipes, second quantitative loop of described substep focalizer is connected with the chromatographic column of described Ultra Performance Liquid Chromatography device by described second high pressure six direction changeover valve, described second high pressure six direction changeover valve comprises two states, load sample shelves (a) and sample introduction shelves (b) respectively, when described second high pressure six direction changeover valve is positioned at load sample shelves (a) state, described second quantitative loop and described chromatographic column isolated, when described second high pressure six direction changeover valve is positioned at sample introduction shelves (b) state, described second quantitative loop is communicated with described chromatographic column.
Above-mentioned substep focuses on the method for combined use of sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that comprising the following steps:
The first step, the state repeatedly switching the second high pressure six direction changeover valve is load sample shelves (a) or sample introduction shelves (b), and described second quantitative loop is carried out substep by described second high pressure six direction changeover valve to described Ultra Performance Liquid Chromatography device and focused on sample introduction;
Second step, when described second high pressure six direction changeover valve is positioned at load sample shelves (a) state, the eluotropic strength changing the mobile phase of Ultra Performance Liquid Chromatography device carries out gradient separations.
A kind of on-line preconcentration-substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that, comprise be connected to each other between two on-line preconcentration device, focus into sampling device and Ultra Performance Liquid Chromatography device step by step,
Described on-line preconcentration device comprises extraction column;
Described Ultra Performance Liquid Chromatography device comprises chromatographic column, high pressure six-way injection valve and the first quantitative loop, and the two ends of described first quantitative loop are connected to high pressure six-way injection valve;
Described substep focalizer comprises the second high pressure six direction changeover valve and the second quantitative loop, the two ends of the second quantitative loop are connected to the second high pressure six direction changeover valve by stainless steel pipes, second quantitative loop of described substep focalizer is connected with the chromatographic column of described Ultra Performance Liquid Chromatography device by described second high pressure six direction changeover valve, described second high pressure six direction changeover valve comprises two states, load sample shelves (a) and sample introduction shelves (b) respectively, when described second high pressure six direction changeover valve is positioned at load sample shelves (a) state, described second quantitative loop and described chromatographic column isolated; When described second high pressure six direction changeover valve is positioned at sample introduction shelves (b) state, described second quantitative loop is communicated with described chromatographic column.
Further, described on-line preconcentration device also comprises the first high pressure six direction changeover valve be connected with described high pressure six-way injection valve, described extraction column one end is connected to described first high pressure six direction changeover valve, the other end focuses into sampling device with described substep and is connected, described first high pressure six direction changeover valve comprises two states, load sample shelves (a) and sample introduction shelves (b) respectively, when described first high pressure six direction changeover valve is positioned at load sample shelves (a) state, described extraction column and described first quantitative loop isolated; Described first high pressure six direction changeover valve be positioned at sample introduction shelves (b) state and described high pressure six-way injection valve is positioned at sample introduction shelves (inject) time, described extraction column is communicated with described first quantitative loop.
Further, described on-line preconcentration device also comprises syringe pump, and described syringe pump is connected with described first high pressure six direction changeover valve, and when described first high pressure six direction changeover valve is positioned at load sample shelves (a) state, described syringe pump is communicated with described extraction column.
Further, described Ultra Performance Liquid Chromatography device also comprises sample introduction needle, detecting device and high pressure liquid phase pump, described high pressure liquid phase pump is connected with described high pressure six-way injection valve, when described high pressure six-way injection valve is in load sample shelves (load), described first quantitative loop and described high pressure liquid phase pump isolated; When described high pressure six-way injection valve is in sample introduction shelves (inject), described first quantitative loop is communicated with described high pressure liquid phase pump.
Further, the second quantitative loop volume that described substep focuses into sampling device is greater than the effluent volume of extraction column.
Above-mentioned on-line preconcentration-substep focuses on the method for combined use of sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that comprising the following steps:
The first step, described first high pressure six direction changeover valve is positioned at sample introduction shelves (b), described second high pressure six direction changeover valve is positioned at sample introduction shelves (b), and described high pressure six-way injection valve is when being positioned at sample introduction shelves (inject), passes into sample solution by described first quantitative loop to described extraction column and extract;
Second step, described first high pressure six direction changeover valve is positioned at load sample shelves (a), described second high pressure six direction changeover valve is positioned at sample introduction shelves (b), and described high pressure six-way injection valve is when being positioned at load sample shelves (load), purification extraction column, injects eluting solvent simultaneously in described first quantitative loop;
3rd step, described first high pressure six direction changeover valve is positioned at sample introduction shelves (b), described second high pressure six direction changeover valve is positioned at load sample shelves (a), and described high pressure six-way injection valve is when being positioned at sample introduction shelves (inject), eluting solvent flows through described extraction column from described first quantitative loop and enters described second quantitative loop;
4th step, described first high pressure six direction changeover valve is positioned at load sample shelves (a), described high pressure six-way injection valve is positioned at load sample shelves (load) simultaneously, being load sample shelves (a) or sample introduction shelves (b) by repeatedly switching the state of the second high pressure six direction changeover valve, carrying out substep to described Ultra Performance Liquid Chromatography device and focusing on sample introduction.
Further, further comprising the steps of:
5th step, described first high pressure six direction changeover valve is positioned at load sample shelves (a), described second high pressure six direction changeover valve is positioned at into load sample shelves (a), when described high pressure six-way injection valve is positioned at load sample shelves (load) simultaneously, the eluotropic strength changing the mobile phase of Ultra Performance Liquid Chromatography device carries out gradient separations.
Further again, described on-line preconcentration-substep focuses on the method for combined use of sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that: control substep by the switching times and frequency controlling the second high pressure six direction changeover valve and focus on the number of times of sample introduction and often walk sampling volume, under certain flow rate, the residence time being positioned at sample introduction shelves by controlling the second high pressure six direction changeover valve controls often to walk sampling volume, by control the second high pressure six direction changeover valve the residence time of load sample shelves control between adjacent two parts substep sample introductions sample between mobile phase volume, determined by the reservation power of total sample volume and analysis thing the interval time often walked between sample injection time and adjacent two steps.
The object of the invention to solve the technical problems also can realize by the following technical solutions.
A kind of on-line preconcentration-the substep proposed according to the present invention focuses on sample introduction-Ultra Performance Liquid Chromatography combined system, and it comprises the following steps: 1) the lower initial flow of Ultra Performance Liquid Chromatography post employing eluotropic strength balances each other chromatographic column; 2) high pressure six-way injection valve is in load sample shelves (load), and sample solution injects the first quantitative loop; 3) high pressure six-way injection valve is in sample introduction shelves (Inject), and under high pressure liquid phase pump drives, sample solution flows through extraction column and carries out enrichment; 4) repeat 2) with 3) step, inject purifying solvent and flow through extraction column and purify; 5) repeat 2) with 3) step, inject eluting solvent and flow through extraction column and carry out wash-out, the second high pressure six direction changeover valve focusing into sampling device (B) by switching substep makes eluent flow into the second quantitative loop; 6) by switching three high pressure six-way valves make the first quantitative loop, the second quantitative loop and extraction column and UHPLC stream isolated; 7) the second high pressure six direction changeover valve of repeated multiple times switching device shifter (B), carries out substep and focuses on sample introduction; 8), after all eluents enter column cap focusing, the eluotropic strength improving mobile phase carries out gradient separations detection.
By technique scheme, on-line preconcentration-substep of the present invention focuses on sample introduction-Ultra Performance Liquid Chromatography combined system and at least has the following advantages:
1) substep focusing sample introduction has stronger spectrum peak compressed capability, can realize on-line concentration and UHPLC on-line coupling, improves degree of separation and the sensitivity of system.
2) system is simple, easily realizes, and only need add high pressure six direction changeover valve, quantitative loop and extraction column on UHPLC instrument basis.
3) design of combined system avoids fiber material to be directly connected with UHPLC chromatographic column, makes it damage from high pressure in elution process, expands the range of choice of fiber material.
4) system to focus on by substep after simplifying and realizes the large volume sample injection analysis of strong solvent sample, avoids peak stretching and distortion, saves the off-line operations such as heavy molten, the high multiple dilutions of evaporate to dryness, improves sensitivity, degree of separation, reduce manual operation.Make large volume sample injection technology can be applied to the fluid sample analyzing thing or matrix poorly water-soluble, this type of sample is difficult to weigh eluotropic strength that is molten or dilution reduction solvent with water.
5) substep focuses into sampling device has good versatility, higher promotional value.Can be used for solving conventional reversed-phase liquid chromatography, capillary chromatography volume overload problem, also can be used as interface in multidimensional liquid chromatography, reduce loss of sensitivity.
More than be illustrated as the general introduction of technical solution of the present invention, in order to can clearer understanding technological means of the present invention, and can be implemented according to the content of instructions, and can become apparent to allow above and other objects of the present invention, feature and advantage, now especially exemplified by preferred embodiment, and coordinate accompanying drawing, be described in detail as follows.
Accompanying drawing explanation
Fig. 1 is that on-line preconcentration-substep provided by the invention focuses on sample introduction-Ultra Performance Liquid Chromatography combined system device (I) and focuses into the schematic diagram of switching repeatedly (II) of sampling device and the second high pressure six direction changeover valve thereof step by step, wherein:
A-on-line preconcentration device;
B-substep focuses into sampling device;
C-Ultra Performance Liquid Chromatography;
1-high pressure six-way injection valve (containing the first quantitative loop); 2-first high pressure six direction changeover valve; 3-second high pressure six direction changeover valve; 4-extraction column; 5-second quantitative loop; 6-chromatographic column; 7-detecting device; 8-high pressure liquid phase pump; 9-syringe pump; 10-initial flow phase; 11-eluate sample.
Fig. 2 is the chromatogram that on-line preconcentration-substep focuses on that sample introduction-Ultra Performance Liquid Chromatography combined system analyzes triazine herbicide mark-on water sample from the beginning, wherein:
A is the chromatogram adopting on-line preconcentration-substep to focus on sample introduction-Ultra Performance Liquid Chromatography combined system analysis;
B analyzes chromatogram for adopting on-line preconcentration-Ultra Performance Liquid Chromatography combined system;
C is Ultra Performance Liquid Chromatography direct injected 50 μ L mark-on water sample chromatogram;
1-Simanex; 2-cyanatryn; 3-symetryne; 4-ametryn; 5-puts out only; 6-Garagard; 7-prometryn; 8-terbutryn; Spiked levels 1 μ g/L, sample volume 4mL.
Fig. 3 is that Ultra Performance Liquid Chromatography disposable sample introduction 320 μ L acetonitrile sample focuses on sample introduction contrast colors spectrogram with substep, wherein,
A is conventional disposable sample introduction chromatogram;
B is that substep focuses on sample introduction chromatogram;
1-nitrobenzene; 2-toluene; 3-o-dichlorobenzene; 4-butyl phthalate; 5-benzofluoranthrene; 6-octyl phthalate; Six kinds of compound concentrations are 1mg/L.
Embodiment
For further setting forth the present invention with the technological means reached predetermined goal of the invention and take and technique effect thereof, below in conjunction with embodiment and accompanying drawing, the structure of sample introduction-Ultra Performance Liquid Chromatography combined system, feature and embodiment thereof are focused on to on-line preconcentration-substep that the present invention proposes, is described in detail as follows.
As shown in Fig. 1 (I), for on-line preconcentration-substep provided by the invention focuses on sample introduction-Ultra Performance Liquid Chromatography combined system device, it comprises: on-line preconcentration device A, substep focus into sampling device B and Ultra Performance Liquid Chromatography device C.First high pressure six direction changeover valve 2 and the second high pressure six direction changeover valve 3 are designated as load sample shelves a by the connected state of solid line position, are designated as sample introduction shelves b by the connected state of dotted line position.
On-line preconcentration device A comprises extraction column 4, first high pressure six direction changeover valve 2 and syringe pump 9.When the first high pressure six direction changeover valve 2 is in a shelves, extraction column 4 is isolated with high pressure liquid phase pump 8, now can be driven by syringe pump 9 and carry out the operation of cleaning, balancing extraction column; When the first high pressure six direction changeover valve 2 is in b shelves, extraction column 4 is connected with high pressure liquid phase pump 8, can carry out extracting under this pump drives, elution action.
Substep focuses into sampling device B and comprises the second high pressure six direction changeover valve 3 and the second quantitative loop 5.When the second high pressure six direction changeover valve 3 is in a shelves, the second quantitative loop 5 is isolated with chromatographic column 6, and now can carry out load sample operation, namely eluent flows into the second quantitative loop; When the second high pressure six direction changeover valve 3 is in b shelves, the second quantitative loop 5 is connected with chromatographic column 6, and now sample flows into chromatographic column 6, belongs to sample introduction state; When the second high pressure six direction changeover valve 3 repeated multiple times switch between a shelves and b shelves, sample in quantitative loop 5 can enter chromatographic column 6 in piecewise, carries out substep focusing.
Ultra Performance Liquid Chromatography device C comprises high pressure liquid phase pump 8, high pressure six-way injection valve 1(containing the first quantitative loop), chromatographic column 6 and detecting device 7.When high pressure six-way injection valve 1 is in Load shelves (load sample shelves), its first quantitative loop and high pressure liquid phase pump 8 isolated, now can inject sample solution, eluting solvent or purifying solvent by sample introduction needle; When high pressure six-way injection valve 1 is in Inject shelves (sample introduction shelves), the first quantitative loop is connected with high pressure liquid phase pump 8, under this pump drives, in ring solution (sample solution, elution volume or purifying solvent) flow through that extraction column carries out extracting, wash-out or purification run.
Embodiment 1: employing on-line preconcentration-substep focuses on the triazine herbicide in sample introduction-Ultra Performance Liquid Chromatography combined system analysis water-like.
Experimental drug, instrument and condition: sample is the mark-on tap water of 8 kinds of triazine herbicides, use Ultra Performance Liquid Chromatography instrument and C18 reverse-phase chromatographic column (specification 3.0 × 100mm, particle diameter 2.2 μm), initial flow is 5% acetonitrile mutually, 0.5min is kept after sample introduction, then in 11.5min, be increased to 38%, finally bring up to 100% maintenance 3min; Flow velocity 1mL/min, extraction column (Inertsil ODS-SP column, 4.0 × 10.0mm, 5 μm).
On-line preconcentration-substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system as shown in Figure 1, the position of operation steps and respective valve is as shown in table 1, often walk concrete operations as follows: 1) 4mL tap water sample injects the first quantitative loop of high pressure six-way injection valve 1 by sample introduction needle, simultaneously 5% acetonitrile balance chromatographic column; 2) under high pressure liquid phase pump 8 drives, sample is pushed extraction column 4 by mobile phase with 1mL/min flow velocity from the first quantitative loop of high pressure six-way injection valve 1 and extracts; 3) distilled water is pumped into extraction column 4 and purifies by syringe pump 9, and eluting solvent (400 μ L acetonitrile) injects the first quantitative loop of high pressure six-way injection valve 1 by sample introduction needle simultaneously; 4) under high pressure liquid phase pump 8 drives, eluting solvent enters the second quantitative loop 5 after the first quantitative loop of six-way injection valve 1 is flow through extraction column 4 by mobile phase with the flow velocity of 1mL/min; 5) repeatedly switch the second high pressure six direction changeover valve 3 to carry out substep and focus on sample introduction, totally 50 times, stop 2s, b shelves at a shelves at every turn and stop 1s; 6) the acetonitrile ratio improving mobile phase carries out gradient separations and detection.
Table 1 on-line preconcentration-substep focuses on the analytical procedure of sample introduction-Ultra Performance Liquid Chromatography combined system and corresponding high pressure six-way valve state
Fig. 2 is mark-on water sample analysis chromatogram from the beginning, and complete, quick wash-out needs 400 μ L acetonitriles as eluting solvent, easily causes the volume overload of Ultra Performance Liquid Chromatography post.Because substep focuses on the effect of sample introduction, on-line coupling analysis can obtain good peak shape (a).If adopt conventional coupling mode, namely eluent focuses on without substep and directly enters chromatographic column, there will be serious bands of a spectrum distortion (b), illustrate that substep focuses on interface arrangement and has powerful bands of a spectrum compression function, effectively can solve the volume overload problem of on-line coupling.A (), compared with (c), illustrates that on-line preconcentration can greatly improve UHPLC sensitivity for analysis.On-line coupling methods analyst 8 kinds of triazine herbicides, detection limit is not higher than 0.011 μ g/L, good in the linearly of 0.08-20.0 μ g/L, RSD is less than 4.0%, the recovery, between 84.3-107.9%, illustrates that the on-line system of use substep focusing sample introduction interface is quick, sensitive, stable and reliable.
Embodiment 2: Ultra Performance Liquid Chromatography substep focuses on large volume sample injection method and analyzes six kinds containing phenyl ring pollutant.
Experimental drug, instrument and condition: sample is the acetonitrile solution containing nitrobenzene, toluene, o-dichlorobenzene, butyl phthalate, benzofluoranthrene and octyl phthalate, use Ultra Performance Liquid Chromatography instrument and C18 reverse-phase chromatographic column (specification 3.0 × 100mm, particle diameter 2.2 μm), employing high pressure six-way injection valve, 1mL stainless steel quantitative loop realize multistep and focus on sample introduction, initial flow is 5% acetonitrile mutually, flow velocity 1mL/min.
It is the simplification that on-line preconcentration-substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system that Ultra Performance Liquid Chromatography substep focuses on large volume sample injection device, as shown in Fig. 1 (I), shed on-line preconcentration device A, make substep focus into sampling device B and be directly connected with Ultra Performance Liquid Chromatography device C.Concrete operations are: use sample introduction needle to inject 320 μ L samples in the second quantitative loop 5 from the second high pressure six direction changeover valve 3.It is repeatedly switch the second high pressure six-way injection valve 3 totally 32 times that substep focuses on sampling condition, stops 2s at load sample shelves at every turn, stops 1s at sample introduction shelves; Conventional disposable sampling condition is switchback load sample shelves after switching second high pressure six direction changeover valve 3 to sample introduction shelves stop 30s.Sample introduction terminates acetonitrile ratio in rear raising mobile phase and carries out gradient separations detection.
In Fig. 3, conventional sample introduction (a) and substep focus on the chromatogram contrast of sample introduction (b), because sampling volume is larger, Ultra Performance Liquid Chromatography post is thinner and short, conventional sample introduction there will be serious peak stretching and distortion (see a), adopt multistep focusing sample introduction can obtain sharp bands (see b), to avoid strong solvent (acetonitrile) to continue to enter chromatographic column in large quantities this is because substep focuses on, thus reduce the solvent effect of sample.Sample injection method of the present invention significantly improves degree of separation and sensitivity.
The above is only preferred embodiment of the present invention, not does any pro forma restriction to the present invention.Although the present invention illustrates as above with preferred embodiment, but and be not used to limit the present invention, any those skilled in the art, do not departing within the scope of technical solution of the present invention, the Equivalent embodiments changing or modify equivalent variations is made when the technology contents of above-mentioned announcement can be utilized, in every case be the content not departing from technical solution of the present invention, the any simple modification done above embodiment according to technical spirit of the present invention, equivalent variations and modification, all still belong in the scope of technical solution of the present invention.
Claims (8)
1. substep focuses on a method for combined use for sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that, described substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system and comprises:
The substep be connected to each other focuses into sampling device and Ultra Performance Liquid Chromatography device,
Described Ultra Performance Liquid Chromatography device comprises chromatographic column;
Described substep focuses into sampling device and comprises the second high pressure six direction changeover valve and the second quantitative loop, the two ends of the second quantitative loop are connected to the second high pressure six direction changeover valve by stainless steel pipes, second quantitative loop of described substep focalizer is connected with the chromatographic column of described Ultra Performance Liquid Chromatography device by described second high pressure six direction changeover valve, described second high pressure six direction changeover valve comprises two states, load sample shelves (a) and sample introduction shelves (b) respectively, when described second high pressure six direction changeover valve is positioned at load sample shelves (a) state, described second quantitative loop and described chromatographic column isolated, when described second high pressure six direction changeover valve is positioned at sample introduction shelves (b) state, described second quantitative loop is communicated with described chromatographic column,
Described method for combined use comprises the following steps:
The first step, the state of repeated multiple times switching second high pressure six direction changeover valve is load sample shelves (a) or sample introduction shelves (b), and described second quantitative loop is carried out substep by described second high pressure six direction changeover valve to described Ultra Performance Liquid Chromatography device and focused on sample introduction;
Second step, when described second high pressure six direction changeover valve is positioned at load sample shelves (a) state, the eluotropic strength changing the mobile phase of Ultra Performance Liquid Chromatography device carries out gradient separations.
2. on-line preconcentration-substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that, comprise be connected to each other between two on-line preconcentration device, substep focus into sampling device and Ultra Performance Liquid Chromatography device,
Described on-line preconcentration device comprises extraction column, the first high pressure six direction changeover valve;
Described Ultra Performance Liquid Chromatography device comprises chromatographic column, high pressure six-way injection valve and the first quantitative loop, and the two ends of described first quantitative loop are connected to high pressure six-way injection valve;
Described substep focalizer comprises the second high pressure six direction changeover valve and the second quantitative loop, the two ends of the second quantitative loop are connected to the second high pressure six direction changeover valve by stainless steel pipes, second quantitative loop of described substep focalizer is connected with the chromatographic column of described Ultra Performance Liquid Chromatography device by described second high pressure six direction changeover valve, described second high pressure six direction changeover valve comprises two states, load sample shelves (a) and sample introduction shelves (b) respectively, when described second high pressure six direction changeover valve is positioned at load sample shelves (a) state, described second quantitative loop and described chromatographic column isolated, when described second high pressure six direction changeover valve is positioned at sample introduction shelves (b) state, described second quantitative loop is communicated with described chromatographic column,
Described high pressure six-way injection valve is connected with the first high pressure six direction changeover valve; Described extraction column one end is connected to described first high pressure six direction changeover valve, and the second high-pressure spray direction changeover valve that the other end focuses into sampling device with substep is connected; Described first high pressure six direction changeover valve comprises two states, load sample shelves (a) and sample introduction shelves (b) respectively, when described first high pressure six direction changeover valve is positioned at load sample shelves (a) state, described extraction column and described first quantitative loop isolated; Described first high pressure six direction changeover valve be positioned at sample introduction shelves (b) state and described high pressure six-way injection valve is positioned at sample introduction shelves (inject) time, described extraction column is communicated with described first quantitative loop.
3. on-line preconcentration-substep according to claim 2 focuses on sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that, described on-line preconcentration device also comprises syringe pump, described syringe pump is connected with described first high pressure six direction changeover valve, when described first high pressure six direction changeover valve is positioned at load sample shelves (a) state, described syringe pump is communicated with described extraction column.
4. on-line preconcentration-substep according to claim 2 focuses on sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that, described Ultra Performance Liquid Chromatography device also comprises sample introduction needle, detecting device and high pressure liquid phase pump, described high pressure liquid phase pump is connected with described high pressure six-way injection valve, when described high pressure six-way injection valve is in load sample shelves (load), described first quantitative loop and described high pressure liquid phase pump isolated; When described high pressure six-way injection valve is in sample introduction shelves (inject), described first quantitative loop is communicated with described high pressure liquid phase pump.
5. on-line preconcentration-substep according to claim 2 focuses on sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that: the second quantitative loop volume that described substep focuses into sampling device is greater than the effluent volume of extraction column.
6. focus on the method for combined use of sample introduction-Ultra Performance Liquid Chromatography combined system according to the arbitrary described on-line preconcentration-substep of claim 2 ~ 5, it is characterized in that comprising the following steps:
The first step, described first high pressure six direction changeover valve is positioned at sample introduction shelves (b), described second high pressure six direction changeover valve is positioned at sample introduction shelves (b), and described high pressure six-way injection valve is when being positioned at sample introduction shelves (inject), passes into sample solution by described first quantitative loop to described extraction column and extract;
Second step, described first high pressure six direction changeover valve is positioned at load sample shelves (a), described second high pressure six direction changeover valve is positioned at sample introduction shelves (b), and described high pressure six-way injection valve is when being positioned at load sample shelves (load), purification extraction column, injects eluting solvent simultaneously in described first quantitative loop;
3rd step, described first high pressure six direction changeover valve is positioned at sample introduction shelves (b), described second high pressure six direction changeover valve is positioned at load sample shelves (a), and described high pressure six-way injection valve is when being positioned at sample introduction shelves (inject), eluting solvent flows through described extraction column from described first quantitative loop and carries out wash-out, and eluent enters described second quantitative loop;
4th step, described first high pressure six direction changeover valve is positioned at load sample shelves (a), described high pressure six-way injection valve is positioned at load sample shelves (load) simultaneously, be load sample shelves (a) or sample introduction shelves (b) by the state of repeated multiple times switching second high pressure six direction changeover valve, carry out substep to described Ultra Performance Liquid Chromatography device and focus on sample introduction.
7. on-line preconcentration-substep according to claim 6 focuses on the method for combined use of sample introduction-Ultra Performance Liquid Chromatography combined system, characterized by further comprising following steps:
5th step, described first high pressure six direction changeover valve is positioned at load sample shelves (a), described second high pressure six direction changeover valve is positioned at into load sample shelves (a), when described high pressure six-way injection valve is positioned at load sample shelves (load) simultaneously, the eluotropic strength changing the mobile phase of Ultra Performance Liquid Chromatography device carries out gradient separations.
8. on-line preconcentration-substep according to claim 7 focuses on the method for combined use of sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that: control substep by the switching times and frequency controlling the second high pressure six direction changeover valve and focus on the number of times of sample introduction and often walk sampling volume, under certain flow rate, the residence time being positioned at sample introduction shelves by controlling the second high pressure six direction changeover valve controls often to walk sampling volume, by control the second high pressure six direction changeover valve the residence time of load sample shelves control between adjacent two parts substep sample introductions sample between mobile phase volume, determined by the reservation power of total sample volume and analysis thing the interval time often walked between sample injection time and adjacent two steps.
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| HUE051209T2 (en) * | 2014-10-27 | 2021-03-01 | Hoffmann La Roche | System and methods for two-dimensional rplc-sfc chromatography |
| CN107870219B (en) * | 2016-09-26 | 2020-09-01 | 中国科学院大连化学物理研究所 | Small chemical derivatization device |
| CN106501428B (en) * | 2016-10-24 | 2018-02-23 | 南方医科大学 | A kind of substep focuses on the method for combined use of axial mixed high-efficient liquid chromatographic separation system |
| CN108593810A (en) * | 2018-07-05 | 2018-09-28 | 中国科学院上海有机化学研究所 | A kind of online purification devices and method for liquid chromatogram |
| CN109799230B (en) * | 2019-01-30 | 2021-09-28 | 中山大学 | Rapid detection device and detection method for metal complex and chiral isomer |
| CN114609312A (en) * | 2022-03-22 | 2022-06-10 | 苏州艾迪迈医疗科技有限公司 | Sample introduction structure and method for sample pretreatment device to chromatographic analysis device |
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| US20040235187A1 (en) * | 2003-05-20 | 2004-11-25 | University Of Maryland, Baltimore County | Platform for analysis of liquid samples |
| US7144502B2 (en) * | 2004-06-04 | 2006-12-05 | Johnson & Johnson | Chromatography system with gradient storage and method for operating the same |
| KR100924369B1 (en) * | 2007-12-04 | 2009-10-30 | 고려대학교 산학협력단 | Ultrahigh-pressure cation exchanger dual on-line solid phase extraction/capillary reverse-phase liquid chromatography system |
| EP2196801B1 (en) * | 2008-12-11 | 2017-08-23 | Spark Holland B.V. | Method and apparatus for injecting a liquid sample in an HPLC analyzing device, and valve assembly for use therein. |
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