CN103134889A - 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
- Publication number
- CN103134889A CN103134889A CN2013100306648A CN201310030664A CN103134889A CN 103134889 A CN103134889 A CN 103134889A CN 2013100306648 A CN2013100306648 A CN 2013100306648A CN 201310030664 A CN201310030664 A CN 201310030664A CN 103134889 A CN103134889 A CN 103134889A
- Authority
- CN
- China
- Prior art keywords
- high pressure
- shelves
- sample
- sample introduction
- changeover valves
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000001195 ultra high performance liquid chromatography Methods 0.000 title abstract description 25
- 238000000034 method Methods 0.000 claims abstract description 16
- 239000000523 sample Substances 0.000 claims description 183
- 238000004704 ultra performance liquid chromatography Methods 0.000 claims description 68
- 238000002347 injection Methods 0.000 claims description 43
- 239000007924 injection Substances 0.000 claims description 43
- 238000000605 extraction Methods 0.000 claims description 35
- 238000004094 preconcentration Methods 0.000 claims description 26
- 238000005070 sampling Methods 0.000 claims description 21
- 239000007791 liquid phase Substances 0.000 claims description 18
- 239000002904 solvent Substances 0.000 claims description 18
- 239000012071 phase Substances 0.000 claims description 15
- 238000000926 separation method Methods 0.000 claims description 14
- 239000003480 eluent Substances 0.000 claims description 8
- 239000012488 sample solution Substances 0.000 claims description 6
- 239000000284 extract Substances 0.000 claims description 5
- 229910001220 stainless steel Inorganic materials 0.000 claims description 5
- 239000010935 stainless steel Substances 0.000 claims description 5
- 239000012491 analyte Substances 0.000 claims description 4
- 238000004458 analytical method Methods 0.000 abstract description 14
- 230000035945 sensitivity Effects 0.000 abstract description 8
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 27
- 238000010168 coupling process Methods 0.000 description 9
- 230000008878 coupling Effects 0.000 description 8
- 238000005859 coupling reaction Methods 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 6
- 238000002470 solid-phase micro-extraction Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
- 238000001228 spectrum Methods 0.000 description 5
- 206010016803 Fluid overload Diseases 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000002657 fibrous material Substances 0.000 description 4
- 239000004009 herbicide Substances 0.000 description 4
- YXFVVABEGXRONW-UHFFFAOYSA-N toluene Substances CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 4
- 230000008901 benefit Effects 0.000 description 3
- LQNUZADURLCDLV-UHFFFAOYSA-N nitrobenzene Chemical compound [O-][N+](=O)C1=CC=CC=C1 LQNUZADURLCDLV-UHFFFAOYSA-N 0.000 description 3
- 238000002414 normal-phase solid-phase extraction Methods 0.000 description 3
- JYEUMXHLPRZUAT-UHFFFAOYSA-N 1,2,3-triazine Chemical compound C1=CN=NN=C1 JYEUMXHLPRZUAT-UHFFFAOYSA-N 0.000 description 2
- RFFLAFLAYFXFSW-UHFFFAOYSA-N 1,2-dichlorobenzene Chemical compound ClC1=CC=CC=C1Cl RFFLAFLAYFXFSW-UHFFFAOYSA-N 0.000 description 2
- DOIRQSBPFJWKBE-UHFFFAOYSA-N dibutyl phthalate Chemical compound CCCCOC(=O)C1=CC=CC=C1C(=O)OCCCC DOIRQSBPFJWKBE-UHFFFAOYSA-N 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 229940000406 drug candidate Drugs 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 239000003777 experimental drug Substances 0.000 description 2
- 239000012530 fluid Substances 0.000 description 2
- 230000002363 herbicidal effect Effects 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000008399 tap water Substances 0.000 description 2
- 235000020679 tap water Nutrition 0.000 description 2
- SVRGUNPFMROZBU-UHFFFAOYSA-N 2-octan-3-yloxycarbonylbenzoic acid Chemical compound CCCCCC(CC)OC(=O)C1=CC=CC=C1C(O)=O SVRGUNPFMROZBU-UHFFFAOYSA-N 0.000 description 1
- MQIUGAXCHLFZKX-UHFFFAOYSA-N Di-n-octyl phthalate Natural products CCCCCCCCOC(=O)C1=CC=CC=C1C(=O)OCCCCCCCC MQIUGAXCHLFZKX-UHFFFAOYSA-N 0.000 description 1
- YZBOVSFWWNVKRJ-UHFFFAOYSA-N Monobutylphthalate Chemical compound CCCCOC(=O)C1=CC=CC=C1C(O)=O YZBOVSFWWNVKRJ-UHFFFAOYSA-N 0.000 description 1
- BJQHLKABXJIVAM-UHFFFAOYSA-N bis(2-ethylhexyl) phthalate Chemical compound CCCCC(CC)COC(=O)C1=CC=CC=C1C(=O)OCC(CC)CCCC BJQHLKABXJIVAM-UHFFFAOYSA-N 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229960002380 dibutyl phthalate Drugs 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000004366 reverse phase liquid chromatography Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000004454 trace mineral analysis Methods 0.000 description 1
Images
Landscapes
- Treatment Of Liquids With Adsorbents In General (AREA)
- Sampling And Sample Adjustment (AREA)
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 a kind of on-line preconcentration-substep and focus on design and analysis and the application of sample introduction-Ultra Performance Liquid Chromatography combined system, be applicable to online pre-service, extraction, desorb, the separation and detection of trace organic substance in the complex samples such as environment, food, medicine, biology.
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 the complex sample extreme trace analysis, usually need to adopt efficient pretreatment technology to purify and enrichment, as Solid-Phase Extraction (SPE) and solid-phase microextraction (SPME).Beneficiation technologies is combined with in the mode of off-line usually with UHPLC at present, and this is that its packing material size is less because the UHPLC chromatographic column is shorter and smaller than conventional liquid-phase chromatographic column, and sampling volume is restricted.Often volume is 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, the UHPLC working pressure is higher, makes not high voltage bearing fiber material be difficult to the on-line coupling with UHPLC.At present still lack the on-line preconcentration analysis that effective strategy is realized UHPLC, for example, the report of SPME and UHPLC on-line coupling do not occur yet.
Summary of the invention
The present invention is intended to easily cause for the eluent of on-line preconcentration the problem of Ultra Performance Liquid Chromatography peak broadening and distortion, design a kind of on-line preconcentration-substep and focus on sample introduction-Ultra Performance Liquid Chromatography combined system, the strategy that focuses on by substep has solved 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, improved degree of separation, analysis efficiency and sensitivity.
Second purpose of the present invention is to fill up present blank without SPME and UHPLC on-line coupling, and the SPME-UHPLC on-line coupled system combines SPME and operates advantage and UHPLC characteristics rapidly and efficiently simple and easy, green low consumption.
The 3rd purpose of the present invention is to focus on by substep the large volume sample injection UHPLC that realizes the strong solvent sample to analyze, the spectrum peak distortion of 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, improved accuracy, and made large volume sample injection method be applicable to the fluid sample of analyte or matrix poorly water-soluble.
The objective of the invention is by realizing 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:
interconnected substep 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 that the second high pressure six direction changeover valves and second quantitatively encircle, the second two ends of quantitatively encircling are connected in the second high pressure six direction changeover valves by stainless steel pipes, second of described substep focalizer quantitatively encircles and is connected with the chromatographic column of described Ultra Performance Liquid Chromatography device by described the second high pressure six direction changeover valves, described the second high pressure six direction changeover valves comprise two states, respectively load sample shelves (a) and sample introduction shelves (b), when described the second high pressure six direction changeover valves are positioned at load sample shelves (a) state, the described second quantitative ring is isolated with described chromatographic column, when described the second high pressure six direction changeover valves were positioned at sample introduction shelves (b) state, the described second quantitative ring was 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 that repeatedly switches the second high pressure six direction changeover valves are load sample shelves (a) or sample introduction shelves (b), and the described second quantitative ring carries out substep by described the second high pressure six direction changeover valves to described Ultra Performance Liquid Chromatography device and focuses on sample introduction;
Second step, when described the second high pressure six direction changeover valves were positioned at load sample shelves (a) state, the eluotropic strength that changes the mobile phase of Ultra Performance Liquid Chromatography device was carried 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, comprises that interconnected on-line preconcentration device, substep focus into sampling device and Ultra Performance Liquid Chromatography device in twos,
Described on-line preconcentration device comprises extraction column;
Described Ultra Performance Liquid Chromatography device comprises that chromatographic column, high pressure six-way injection valve and first quantitatively encircle, and the described first two ends of quantitatively encircling are connected in the high pressure six-way injection valve;
Described substep focalizer comprises that the second high pressure six direction changeover valves and second quantitatively encircle, the second two ends of quantitatively encircling are connected in the second high pressure six direction changeover valves by stainless steel pipes, second of described substep focalizer quantitatively encircles and is connected with the chromatographic column of described Ultra Performance Liquid Chromatography device by described the second high pressure six direction changeover valves, described the second high pressure six direction changeover valves comprise two states, respectively load sample shelves (a) and sample introduction shelves (b), when described the second high pressure six direction changeover valves were positioned at load sample shelves (a) state, the described second quantitative ring was isolated with described chromatographic column; When described the second high pressure six direction changeover valves were positioned at sample introduction shelves (b) state, the described second quantitative ring was communicated with described chromatographic column.
Further, described on-line preconcentration device also comprises the first high pressure six direction changeover valves that are connected with described high pressure six-way injection valve, described extraction column one end is connected in described the first high pressure six direction changeover valves, the other end focuses into sampling device with described substep and is connected, described the first high pressure six direction changeover valves comprise two states, be respectively load sample shelves (a) and sample introduction shelves (b), when described the first high pressure six direction changeover valves were positioned at load sample shelves (a) state, described extraction column and described first quantitatively ring was isolated; When described the first high pressure six direction changeover valves were positioned at sample introduction shelves (b) state and described high pressure six-way injection valve and are positioned at sample introduction shelves (inject), described extraction column and described first quantitatively ring was communicated with.
Further, described on-line preconcentration device also comprises syringe pump, and described syringe pump is connected with described the first high pressure six direction changeover valves, and when described the first high pressure six direction changeover valves were positioned at load sample shelves (a) state, described syringe pump was 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 was in load sample shelves (load), the described first quantitative ring was isolated with described high pressure liquid phase pump; When described high pressure six-way injection valve was in sample introduction shelves (inject), the described first quantitative ring was communicated with described high pressure liquid phase pump.
Further, described substep focuses into second of sampling device and quantitatively encircles volume 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 the first high pressure six direction changeover valves are positioned at sample introduction shelves (b), described the second high pressure six direction changeover valves are positioned at sample introduction shelves (b), and when described high pressure six-way injection valve is positioned at sample introduction shelves (inject), passes into sample solution by the described first described extraction column of quantitative hoop and extract;
Second step, described the first high pressure six direction changeover valves are positioned at load sample shelves (a), described the second high pressure six direction changeover valves are positioned at sample introduction shelves (b), and when described high pressure six-way injection valve is positioned at load sample shelves (load), purify extraction column, quantitatively inject eluting solvent in ring to described first simultaneously;
The 3rd step, described the first high pressure six direction changeover valves are positioned at sample introduction shelves (b), described the second high pressure six direction changeover valves are positioned at load sample shelves (a), and when described high pressure six-way injection valve was positioned at sample introduction shelves (inject), eluting solvent was crossed described extraction column and is entered described second from the described first quantitative circulation and quantitatively encircles;
The 4th step, described the first high pressure six direction changeover valves are 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 that repeatedly switches the second high pressure six direction changeover valves, carry out substep to described Ultra Performance Liquid Chromatography device and focus on sample introduction.
Further, further comprising the steps of:
The 5th step, described the first high pressure six direction changeover valves are positioned at load sample shelves (a), described the second high pressure six direction changeover valves are positioned at into load sample shelves (a), when described high pressure six-way injection valve was positioned at load sample shelves (load) simultaneously, the eluotropic strength that changes the mobile phase of Ultra Performance Liquid Chromatography device was carried 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 by switching times and the frequency of controlling the second high pressure six direction changeover valves number of times and the every stepping sample volume that substep focuses on sample introduction, under certain flow rate, the residence time that is positioned at the sample introduction shelves by controlling the second high pressure six direction changeover valves is controlled every stepping sample volume, by control the second high pressure six direction changeover valves the residence time of load sample shelves control between adjacent two parts the substep sample introductions sample between mobile phase volume, determine by the reservation of total sample volume and analyte is strong and weak the interval time between every stepping sample 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 that proposes according to the present invention-substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system, and it comprises the following steps: 1) the lower initial flow of the Ultra Performance Liquid Chromatography post employing eluotropic strength chromatographic column that balances each other; 2) the high pressure six-way injection valve is in load sample shelves (load), and sample solution injects first and quantitatively encircles; 3) the high pressure six-way injection valve is in sample introduction shelves (Inject), and the sample solution extraction column of flowing through carries out enrichment under the high pressure liquid phase pump drives; 4) repeat 2) with 3) step, inject purifying solvent and the extraction column of flowing through and purify; 5) repeat 2) with 3) step, inject eluting solvent and the extraction column of flowing through and carry out wash-out, the second high pressure six direction changeover valves that focus into sampling device (B) by switching substep make eluent flow into the second quantitative ring; 6) make first quantitatively ring, the second quantitative ring and extraction column and UHPLC stream are isolated by switching three high pressure six-way valves; 7) the second high pressure six direction changeover valves of repeated multiple times switching device shifter (B), carry out substep and focus on sample introduction; 8) after all eluents enter column cap focusing, improve the eluotropic strength of mobile phase and carry out the gradient separations detection.
By technique scheme, on-line preconcentration of the present invention-substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system and has the following advantages at least:
1) focusing on sample introduction has stronger spectrum peak pressure contracting ability step by step, 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, only needs to add high pressure six direction changeover valves, quantitatively ring and extraction column on UHPLC instrument basis.
3) design of combined system avoids fiber material directly to be connected with the UHPLC chromatographic column, makes it avoid the high pressure damage in elution process, has enlarged the range of choice of fiber material.
4) can focus on the large volume sample injection analysis that realizes the strong solvent sample by substep after system simplification, avoid peak broadening and distortion, save the off-line operations such as heavy molten, the high multiple dilution of evaporate to dryness, improve sensitivity, degree of separation, reduce manually-operated.Make the large volume sample injection technology can be applied to the fluid sample of analyte or matrix poorly water-soluble, this type of sample is difficult to water and weighs eluotropic strength molten or dilution reduction solvent.
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 and be used for the multidimensional liquid chromatography, reduce loss of sensitivity.
More than be illustrated as the general introduction of technical solution of the present invention, for can clearer understanding technological means of the present invention, and can be implemented according to the content of instructions, and for above and other objects of the present invention, feature and advantage can be become apparent, now especially exemplified by preferred embodiment, and the cooperation accompanying drawing, be described in detail as follows.
Description of drawings
Fig. 1 is that on-line preconcentration provided by the invention-substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system device (I) and substep focuses into the schematic diagram of switching repeatedly (II) of sampling device and the second high pressure six direction changeover valves thereof, wherein:
A-on-line preconcentration device;
B-focuses into sampling device step by step;
The C-Ultra Performance Liquid Chromatography;
1-high pressure six-way injection valve (containing first quantitatively encircles); 2-the first high pressure six direction changeover valves; 3-the second high pressure six direction changeover valves; The 4-extraction column; 5-second quantitatively encircles; The 6-chromatographic column; The 7-detecting device; 8-high pressure liquid phase pump; The 9-syringe pump; 10-initial flow phase; 11-eluent sample.
Fig. 2 is that on-line preconcentration-substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system analysis triazine herbicide mark-on chromatogram of water sample from the beginning, wherein:
A is for adopting on-line preconcentration-substep to focus on the chromatogram that sample introduction-Ultra Performance Liquid Chromatography combined system is analyzed;
B is for adopting on-line preconcentration-Ultra Performance Liquid Chromatography combined system to analyze chromatogram;
C is Ultra Performance Liquid Chromatography direct injected 50 μ L mark-on water sample chromatograms;
The 1-Simanex; The 2-cyanatryn; The 3-symetryne; The 4-ametryn; 5-puts out only; The 6-Garagard; The 7-prometryn; The 8-terbutryn; Mark-on concentration 1 μ g/L, sample volume 4mL.
Fig. 3 is that the disposable sample introduction 320 μ L acetonitrile samples of Ultra Performance Liquid Chromatography focus on sample introduction contrast colors spectrogram with substep, wherein,
A is conventional disposable sample introduction chromatogram;
B focuses on the sample introduction chromatogram for substep;
1-nitrobenzene; 2-toluene; The 3-o-dichlorobenzene; The 4-butyl phthalate; The 5-benzofluoranthrene; The 6-octyl phthalate; Six kinds of compound concentrations are 1mg/L.
Embodiment
Technological means and the technique effect thereof taked to reach predetermined goal of the invention for further setting forth the present invention, below in conjunction with embodiment and accompanying drawing, the on-line preconcentration that the present invention is proposed-substep focuses on structure, feature and the embodiment thereof of sample introduction-Ultra Performance Liquid Chromatography combined system, is described in detail as follows.
As shown in Fig. 1 (I), for on-line preconcentration provided by the invention-substep 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.The first high pressure six direction changeover valves 2 and the second high pressure six direction changeover valves 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, the first high pressure six direction changeover valves 2 and syringe pump 9.When the first high pressure six direction changeover valves 2 were in a shelves, extraction column 4 was isolated with high pressure liquid phase pump 8, this moment can by syringe pump 9 drive clean, the operation of balance extraction column; When the first high pressure six direction changeover valves 2 were in the b shelves, extraction column 4 was connected with high pressure liquid phase pump 8, can extract under this pump drives, the wash-out operation.
Substep focuses into sampling device B and comprises quantitatively ring 5 of the second high pressure six direction changeover valves 3 and second.When the second high pressure six direction changeover valves 3 were in a shelves, the second quantitative ring 5 was isolated with chromatographic column 6, can carry out the load sample operation this moment, i.e. eluent inflow second is ring quantitatively; When the second high pressure six direction changeover valves 3 were in the b shelves, second quantitatively encircles 5 was connected with chromatographic column 6, and this moment, sample flowed into chromatographic column 6, belonged to the sample introduction state; When the second high pressure six direction changeover valves 3 switch between a shelves and b shelves repeated multiple timesly, the sample that quantitatively encircles in 5 can enter chromatographic column 6 in the piecewise, carries out substep focusing.
Ultra Performance Liquid Chromatography device C comprises that high pressure liquid phase pump 8, high pressure six-way injection valve 1(contain the first quantitative ring), chromatographic column 6 and detecting device 7.When high pressure six-way injection valve 1 was in Load shelves (load sample shelves), its first quantitative ring was isolated with high pressure liquid phase pump 8, and this moment, available sample introduction needle was injected sample solution, eluting solvent or purifying solvent; When high pressure six-way injection valve 1 is in Inject shelves (sample introduction shelves), first quantitatively ring be connected with high pressure liquid phase pump 8, under this pump driving, the extraction column of flowing through extracts solution in ring (sample solution, elution volume or purifying solvent), 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 μ m), initial flow is 5% acetonitrile mutually, keep 0.5min after sample introduction, then be increased to 38% in 11.5min, bring up at last 100% and keep 3min; Flow velocity 1mL/min, extraction column (Inertsil ODS-SP column, 4.0 * 10.0mm, 5 μ m).
On-line preconcentration-substep focuses on that sample introduction-the 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, per step concrete operations are as follows: 1) 4mL tap water sample injects the first quantitatively ring, simultaneously 5% acetonitrile balance chromatographic column of high pressure six-way injection valve 1 by sample introduction needle; 2) under high pressure liquid phase pump 8 drives, sample from high pressure six-way injection valve 1 first quantitatively ring pushed extraction column 4 by mobile phase with the 1mL/min flow velocity and extract; 3) syringe pump 9 pumps into extraction column 4 purifications with distilled water, and while eluting solvent (400 μ L acetonitrile) injects first of high pressure six-way injection valve 1 by sample introduction needle and quantitatively encircles; 4) under high pressure liquid phase pump 8 drives, eluting solvent from six-way injection valve 1 first quantitatively ring flow through with the flow velocity of 1mL/min by mobile phase and enter the second quantitative ring 5 after extraction column 4; 5) repeatedly switch the second high pressure six direction changeover valves 3 and carry out substep and focus on sample introduction, totally 50 times, stop 2s at a shelves at every turn, the b shelves stop 1s; 6) the acetonitrile ratio that improves mobile phase is carried out gradient separations and detection.
Table 1 on-line preconcentration-substep focuses on analytical procedure and the corresponding high pressure six-way valve state of sample introduction-Ultra Performance Liquid Chromatography combined system
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.Focus on the effect of sample introduction due to substep, the on-line coupling analysis can obtain good peak shape (a).If adopt conventional coupling mode, be that eluent directly enters chromatographic column without substep focusing, serious bands of a spectrum distortion (b) can occur, illustrate that substep focuses on interface arrangement and has powerful bands of a spectrum compression function, can effectively solve the volume overload problem of on-line coupling.(a) compare with (c), illustrate that on-line preconcentration can greatly improve the UHPLC sensitivity for analysis.8 kinds of triazine herbicides of on-line coupling methods analyst, detection limit is not higher than 0.011 μ g/L, linear good between 0.08-20.0 μ g/L, RSD is less than 4.0%, the recovery illustrates that the on-line system that uses substep to focus on the sample introduction interface is quick, sensitive, stable and reliable between 84.3-107.9%.
Embodiment 2: Ultra Performance Liquid Chromatography focuses on step by step six kinds of large volume sample injection method analyses and contains the phenyl ring pollutant.
Experimental drug, instrument and condition: sample is the acetonitrile solution that contains 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 μ m), adopt high pressure six-way injection valve, 1mL stainless steel quantitatively to encircle and realize that multistep focuses 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 focuses on the large volume sample injection device step by step, as shown in Fig. 1 (I), shed on-line preconcentration device A, make substep focus into sampling device B and directly be connected with Ultra Performance Liquid Chromatography device C and get final product.Concrete operations are: use sample introduction needle to inject 320 μ L samples from the second high pressure six direction changeover valves 3 and quantitatively encircle 5 in second.It is repeatedly to switch the second high pressure six-way injection valve 3 totally 32 times that substep focuses on sampling condition, stops 2s at the load sample shelves at every turn, stops 1s at the sample introduction shelves; Conventional disposable sampling condition is to switch the second high pressure six direction changeover valves 3 to sample introduction shelves to stop switchback load sample shelves after 30s.In the rear raising of sample introduction end mobile phase, the acetonitrile ratio is carried out the 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, the Ultra Performance Liquid Chromatography post is thinner and short, serious peak broadening and distortion can appear in conventional sample introduction (to be seen a), adopt multistep to focus on sample introduction and can obtain sharp-pointed bands of a spectrum (seeing b), this is to be avoided strong solvent (acetonitrile) to continue to enter in large quantities chromatographic column because substep focuses on, thereby reduces the solvent effect of sample.Sample injection method of the present invention has significantly improved degree of separation and sensitivity.
The above is only preferred embodiment of the present invention, is not the present invention is done any pro forma restriction.Although the present invention with the preferred embodiment explanation as above, yet be not to limit the present invention, any those skilled in the art, within not breaking away from the technical solution of the present invention scope, make the equivalent embodiment that changes or modify equivalent variations when the technology contents that can utilize above-mentioned announcement, in every case be the content that does not break away from technical solution of the present invention,, all still belong in the scope of technical solution of the present invention any simple modification, equivalent variations and modification that above embodiment does according to technical spirit of the present invention.
Claims (10)
1. a substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that comprising:
Interconnected substep 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 that the second high pressure six direction changeover valves and second quantitatively encircle, the second two ends of quantitatively encircling are connected in the second high pressure six direction changeover valves by stainless steel pipes, second of described substep focalizer quantitatively encircles and is connected with the chromatographic column of described Ultra Performance Liquid Chromatography device by described the second high pressure six direction changeover valves, described the second high pressure six direction changeover valves comprise two states, respectively load sample shelves (a) and sample introduction shelves (b), when described the second high pressure six direction changeover valves are positioned at load sample shelves (a) state, the described second quantitative ring is isolated with described chromatographic column, when described the second high pressure six direction changeover valves were positioned at sample introduction shelves (b) state, the described second quantitative ring was communicated with described chromatographic column.
2. substep according to claim 1 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 of repeated multiple times switching the second high pressure six direction changeover valves are load sample shelves (a) or sample introduction shelves (b), and the described second quantitative ring carries out substep by described the second high pressure six direction changeover valves to described Ultra Performance Liquid Chromatography device and focuses on sample introduction;
Second step, when described the second high pressure six direction changeover valves were positioned at load sample shelves (a) state, the eluotropic strength that changes the mobile phase of Ultra Performance Liquid Chromatography device was carried out gradient separations.
3. on-line preconcentration-substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that, comprises that interconnected on-line preconcentration device, substep focus into sampling device and Ultra Performance Liquid Chromatography device in twos,
Described on-line preconcentration device comprises extraction column;
Described Ultra Performance Liquid Chromatography device comprises that chromatographic column, high pressure six-way injection valve and first quantitatively encircle, and the described first two ends of quantitatively encircling are connected in the high pressure six-way injection valve;
Described substep focalizer comprises that the second high pressure six direction changeover valves and second quantitatively encircle, the second two ends of quantitatively encircling are connected in the second high pressure six direction changeover valves by stainless steel pipes, second of described substep focalizer quantitatively encircles and is connected with the chromatographic column of described Ultra Performance Liquid Chromatography device by described the second high pressure six direction changeover valves, described the second high pressure six direction changeover valves comprise two states, respectively load sample shelves (a) and sample introduction shelves (b), when described the second high pressure six direction changeover valves were positioned at load sample shelves (a) state, the described second quantitative ring was isolated with described chromatographic column; When described the second high pressure six direction changeover valves were positioned at sample introduction shelves (b) state, the described second quantitative ring was communicated with described chromatographic column.
4. on-line preconcentration according to claim 3-substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that, described on-line preconcentration device also comprises the first high pressure six direction changeover valves that are connected with described high pressure six-way injection valve, described extraction column one end is connected in described the first high pressure six direction changeover valves, the other end focuses into sampling device with described substep and is connected, described the first high pressure six direction changeover valves comprise two states, respectively load sample shelves (a) and sample introduction shelves (b), when described the first high pressure six direction changeover valves are positioned at load sample shelves (a) state, described extraction column and described first quantitatively ring is isolated, when described the first high pressure six direction changeover valves were positioned at sample introduction shelves (b) state and described high pressure six-way injection valve and are positioned at sample introduction shelves (inject), described extraction column and described first quantitatively ring was communicated with.
5. on-line preconcentration according to claim 4-substep 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 the first high pressure six direction changeover valves, when described the first high pressure six direction changeover valves were positioned at load sample shelves (a) state, described syringe pump was communicated with described extraction column.
6. on-line preconcentration according to claim 3-substep 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 was in load sample shelves (load), the described first quantitative ring was isolated with described high pressure liquid phase pump; When described high pressure six-way injection valve was in sample introduction shelves (inject), the described first quantitative ring was communicated with described high pressure liquid phase pump.
7. on-line preconcentration according to claim 3-substep focuses on sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that: described substep focuses into second of sampling device and quantitatively encircles volume greater than the effluent volume of extraction column.
8. according to claim 4～7 arbitrary described on-line preconcentration-substeps focus 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 the first high pressure six direction changeover valves are positioned at sample introduction shelves (b), described the second high pressure six direction changeover valves are positioned at sample introduction shelves (b), and when described high pressure six-way injection valve is positioned at sample introduction shelves (inject), passes into sample solution by the described first described extraction column of quantitative hoop and extract;
Second step, described the first high pressure six direction changeover valves are positioned at load sample shelves (a), described the second high pressure six direction changeover valves are positioned at sample introduction shelves (b), and when described high pressure six-way injection valve is positioned at load sample shelves (load), purify extraction column, quantitatively inject eluting solvent in ring to described first simultaneously;
The 3rd step, described the first high pressure six direction changeover valves are positioned at sample introduction shelves (b), described the second high pressure six direction changeover valves are positioned at load sample shelves (a), and when described high pressure six-way injection valve is positioned at sample introduction shelves (inject), eluting solvent is crossed described extraction column from the described first quantitative circulation and is carried out wash-out, and eluent enters described second and quantitatively encircles;
The 4th step, described the first high pressure six direction changeover valves are positioned at load sample shelves (a), described high pressure six-way injection valve is positioned at load sample shelves (load) simultaneously, state by repeated multiple times switching the second high pressure six direction changeover valves is load sample shelves (a) or sample introduction shelves (b), carries out substep to described Ultra Performance Liquid Chromatography device and focuses on sample introduction.
9. on-line preconcentration according to claim 8-substep focuses on the method for combined use of sample introduction-Ultra Performance Liquid Chromatography combined system, characterized by further comprising following steps:
The 5th step, described the first high pressure six direction changeover valves are positioned at load sample shelves (a), described the second high pressure six direction changeover valves are positioned at into load sample shelves (a), when described high pressure six-way injection valve was positioned at load sample shelves (load) simultaneously, the eluotropic strength that changes the mobile phase of Ultra Performance Liquid Chromatography device was carried out gradient separations.
10. on-line preconcentration according to claim 9-substep focuses on the method for combined use of sample introduction-Ultra Performance Liquid Chromatography combined system, it is characterized in that: control by switching times and the frequency of controlling the second high pressure six direction changeover valves number of times and the every stepping sample volume that substep focuses on sample introduction, under certain flow rate, the residence time that is positioned at the sample introduction shelves by controlling the second high pressure six direction changeover valves is controlled every stepping sample volume, by control the second high pressure six direction changeover valves the residence time of load sample shelves control between adjacent two parts the substep sample introductions sample between mobile phase volume, determine by the reservation of total sample volume and analyte is strong and weak the interval time between every stepping sample time and adjacent two steps.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310030664.8A CN103134889B (en) | 2013-01-25 | 2013-01-25 | On-line enrichment-substep focus sample introduction-ultra-high performance liquid chromatography combination system and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310030664.8A CN103134889B (en) | 2013-01-25 | 2013-01-25 | On-line enrichment-substep focus sample introduction-ultra-high performance liquid chromatography combination system and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103134889A true CN103134889A (en) | 2013-06-05 |
| CN103134889B CN103134889B (en) | 2015-03-04 |
Family
ID=48494993
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310030664.8A Expired - Fee Related CN103134889B (en) | 2013-01-25 | 2013-01-25 | On-line enrichment-substep focus sample introduction-ultra-high performance liquid chromatography combination system and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103134889B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106501428A (en) * | 2016-10-24 | 2017-03-15 | 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 |
| CN107870219A (en) * | 2016-09-26 | 2018-04-03 | 中国科学院大连化学物理研究所 | A kind of small chemical derivatization device |
| 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 |
| CN109799230A (en) * | 2019-01-30 | 2019-05-24 | 中山大学 | The device for fast detecting and detection method of a kind of metal complex and chiral isomer |
| CN110514770A (en) * | 2014-10-27 | 2019-11-29 | 豪夫迈·罗氏有限公司 | For two to RPLC-SFC chromatography system and method |
| 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 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050269264A1 (en) * | 2004-06-04 | 2005-12-08 | Adam Fermier | Chromatography system with gradient storage and method for operating the same |
| KR20090058287A (en) * | 2007-12-04 | 2009-06-09 | 고려대학교 산학협력단 | Ultrahigh-pressure cation exchanger dual on-line solid phase extraction/capillary reverse-phase liquid chromatography system |
| US20090200181A1 (en) * | 2003-05-20 | 2009-08-13 | University Of Maryland, Baltimore County | Platform for analysis liquid samples |
| EP2196801A1 (en) * | 2008-12-11 | 2010-06-16 | Spark Holland B.V. | Method and apparatus for injecting a liquid sample in an HPLC analyzing device, and valve assembly for use therein. |
-
2013
- 2013-01-25 CN CN201310030664.8A patent/CN103134889B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090200181A1 (en) * | 2003-05-20 | 2009-08-13 | University Of Maryland, Baltimore County | Platform for analysis liquid samples |
| US20050269264A1 (en) * | 2004-06-04 | 2005-12-08 | Adam Fermier | Chromatography system with gradient storage and method for operating the same |
| KR20090058287A (en) * | 2007-12-04 | 2009-06-09 | 고려대학교 산학협력단 | Ultrahigh-pressure cation exchanger dual on-line solid phase extraction/capillary reverse-phase liquid chromatography system |
| EP2196801A1 (en) * | 2008-12-11 | 2010-06-16 | Spark Holland B.V. | Method and apparatus for injecting a liquid sample in an HPLC analyzing device, and valve assembly for use therein. |
Non-Patent Citations (1)
| Title |
|---|
| 钟启升等: "样品前处理-色谱分析在线联用技术的研究进展", 《色谱》 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110514770A (en) * | 2014-10-27 | 2019-11-29 | 豪夫迈·罗氏有限公司 | For two to RPLC-SFC chromatography system and method |
| CN107870219A (en) * | 2016-09-26 | 2018-04-03 | 中国科学院大连化学物理研究所 | A kind of small chemical derivatization device |
| CN106501428A (en) * | 2016-10-24 | 2017-03-15 | 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 |
| 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 |
| CN109799230A (en) * | 2019-01-30 | 2019-05-24 | 中山大学 | The device for fast detecting and detection method of a kind of metal complex and chiral isomer |
| 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 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103134889B (en) | 2015-03-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103940941B (en) | Automatic pre-treatment-Ultra Performance Liquid Chromatography/mass spectrum on-line analysis system | |
| CN103134889B (en) | On-line enrichment-substep focus sample introduction-ultra-high performance liquid chromatography combination system and application | |
| Clavijo et al. | Analytical strategies for coupling separation and flow-injection techniques | |
| CN100399021C (en) | Liquid chromatograph | |
| US9679757B2 (en) | Liquid chromatography systems and methods | |
| CN107589190B (en) | Large-volume sample introduction-double solid phase extraction-high performance liquid chromatography online combined equipment | |
| US9470664B2 (en) | Chromatographic interface | |
| CN103930778A (en) | Valve and splitting system for multi-dimensional liquid analysis | |
| EP2807410A2 (en) | Liquid sampling valve | |
| CN101169391B (en) | Two-dimensional highly effective liquid phase chromatographic system and its uses | |
| KR101574272B1 (en) | Two dimensional liquid chromatography system for Heart-cut method | |
| CN1828290A (en) | Method for analyzing trace organic substance in water using on-line combined solid phase extraction and liquid chromatography | |
| CN104422735A (en) | Preparative circulating chromatography apparatus | |
| CN110286174A (en) | A kind of online highly effective liquid phase chromatographic device of multidimensional | |
| JP6260719B2 (en) | Liquid chromatograph | |
| Yang et al. | On-column and gradient focusing-induced high-resolution separation in narrow open tubular liquid chromatography and a simple and economic approach for pico-gradient separation | |
| CN101762666A (en) | Multidimensional chromatography combination separation device | |
| CN104076113A (en) | Online hydrophilic interaction chromatography/reversed phase chromatography serial interface device and application thereof | |
| CN220584136U (en) | Multidimensional multichannel liquid chromatographic separation system | |
| CN201348625Y (en) | Multidimensional chromatography separator | |
| CN202393749U (en) | System for preparing liquid chromatogram | |
| CN201464435U (en) | Fully-automatic device used for solid phase micro extraction and liquid chromatogram | |
| CN201223741Y (en) | Small-sized simulated moving bed chromatography device | |
| CN211086206U (en) | Three-dimensional online ultra-high performance liquid chromatograph | |
| CN215415249U (en) | Production type circulating multidimensional liquid chromatography separation system |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150304 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |