CN109959741A - A kind of binary channels on-line analysis pre-treatment and off-line analysis one liquid chromatograph and its application method - Google Patents
A kind of binary channels on-line analysis pre-treatment and off-line analysis one liquid chromatograph and its application method Download PDFInfo
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- 239000007788 liquid Substances 0.000 title claims abstract description 117
- 238000004458 analytical method Methods 0.000 title claims abstract description 72
- 238000000034 method Methods 0.000 title claims abstract description 35
- 238000002203 pretreatment Methods 0.000 title claims abstract description 29
- 238000000746 purification Methods 0.000 claims abstract description 43
- 238000004587 chromatography analysis Methods 0.000 claims abstract description 35
- 238000001212 derivatisation Methods 0.000 claims abstract description 25
- 230000008569 process Effects 0.000 claims abstract description 25
- 238000010828 elution Methods 0.000 claims description 11
- 230000003213 activating effect Effects 0.000 claims description 9
- 230000001172 regenerating effect Effects 0.000 claims description 9
- 239000002699 waste material Substances 0.000 claims description 7
- 239000011159 matrix material Substances 0.000 claims description 4
- 239000012535 impurity Substances 0.000 claims description 3
- 238000007445 Chromatographic isolation Methods 0.000 claims 1
- 239000003086 colorant Substances 0.000 claims 1
- 238000001917 fluorescence detection Methods 0.000 claims 1
- 239000007791 liquid phase Substances 0.000 abstract description 8
- 238000000926 separation method Methods 0.000 abstract description 5
- 239000000126 substance Substances 0.000 abstract description 3
- 230000008859 change Effects 0.000 abstract description 2
- 238000000605 extraction Methods 0.000 abstract 1
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 48
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 27
- 239000012071 phase Substances 0.000 description 19
- IISBACLAFKSPIT-UHFFFAOYSA-N bisphenol A Chemical compound C=1C=C(O)C=CC=1C(C)(C)C1=CC=C(O)C=C1 IISBACLAFKSPIT-UHFFFAOYSA-N 0.000 description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 16
- XWIYFDMXXLINPU-UHFFFAOYSA-N Aflatoxin G Chemical compound O=C1OCCC2=C1C(=O)OC1=C2C(OC)=CC2=C1C1C=COC1O2 XWIYFDMXXLINPU-UHFFFAOYSA-N 0.000 description 11
- 229940106691 bisphenol a Drugs 0.000 description 8
- OQIQSTLJSLGHID-WNWIJWBNSA-N aflatoxin B1 Chemical compound C=1([C@@H]2C=CO[C@@H]2OC=1C=C(C1=2)OC)C=2OC(=O)C2=C1CCC2=O OQIQSTLJSLGHID-WNWIJWBNSA-N 0.000 description 7
- 238000011088 calibration curve Methods 0.000 description 7
- 230000001419 dependent effect Effects 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- 238000007689 inspection Methods 0.000 description 6
- 229930195730 Aflatoxin Natural products 0.000 description 4
- 239000005409 aflatoxin Substances 0.000 description 4
- 239000002115 aflatoxin B1 Substances 0.000 description 4
- 229930020125 aflatoxin-B1 Natural products 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 239000000463 material Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- 230000005284 excitation Effects 0.000 description 2
- 238000011049 filling Methods 0.000 description 2
- 238000012417 linear regression Methods 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000000825 ultraviolet detection Methods 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 229930063498 Aflatoxin G1 Natural products 0.000 description 1
- XWIYFDMXXLINPU-WNWIJWBNSA-N Aflatoxin G1 Chemical compound O=C1OCCC2=C1C(=O)OC1=C2C(OC)=CC2=C1[C@@H]1C=CO[C@@H]1O2 XWIYFDMXXLINPU-WNWIJWBNSA-N 0.000 description 1
- 241000228197 Aspergillus flavus Species 0.000 description 1
- 238000001994 activation Methods 0.000 description 1
- 239000002098 aflatoxin G1 Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 238000007781 pre-processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/24—Automatic injection systems
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses a kind of binary channels on-line analysis pre-treatment and off-line analysis one liquid chromatograph and its application methods, including autosampler, two chromatogram pumps, three six-way valves, in-line purification column, two chromatography columns, Photochemical derivatization device, UV detector and fluorescence detector, can switch between off-line analysis and online preceding processing analysis both of which according to the needs of use.According to the online analysis of shift channel of the property of sample to be analysed, substance of different nature is made to realize offline or online separation analysis in different chromatographic columns, solves the disadvantage that Traditional liquid phase chromatograph can only do off-line analysis or in-line purification analysis.When being used as online pre-treatment analysis, sample can directly detect after simple extraction without pre-treatment, greatly shorten testing process, reduce manual operation, reduce analysis cost.Make not needing to change system when off-line analysis, directly switches, effectively improve the service efficiency and analysis ability of instrument.
Description
Technical field
The present invention relates to liquid chromatographs, more particularly to a kind of binary channels on-line analysis pre-treatment and off-line analysis one
Liquid chromatograph and its application method.
Background technique
Liquid chromatogram is a kind of common separation and detection device, but in the application it has been found that some problems, than
If pretreatment process in liquid chromatography detection is complicated, time-consuming and laborious, the material cost of pre-treatment is very high.And it establishes a set of online
Preprocessing system, sample through simply extract after can sample introduction, operate it is very easy, manpower is greatly saved, can effectively solve sample
Product amount is big, the problem of being short of hands.Importantly, since in-line purification column only carries out only a small amount of sample for entering system
Change, purge amount greatly reduces, and can be greatly saved experimental cost.
Although online pre-treatment liquid phase chromatographic band has carried out the saving in cost, we still face new problem --- nothing
Method carries out offline inspection, if sample is not suitable for online pre-treatment or when sample substrate simply only needs Conventional Off-line to analyze, or
When person needs the chromatography column of other different separation mechanisms, it is necessary to which system flow path is reverted to initial state.In face of sample
Inspected number is big, and the more situation of sample type has dealt with larger difficulty.Often switch in two kinds of system modes, operates
It is troublesome, it is also unfavorable to the service life of instrument, and also disassembly system is professional very strong, can not operate without trained personnel.
If this defect of liquid chromatogram can be directed to, offline liquid chromatogram point can either be completed by realizing on a liquid chromatograph
Analysis, and on-line chromatograph analysis can be completed, realize the online switching of a variety of liquid chromatographic systems, online and offline system can also
Different chromatographic columns is selected according to the property of measured object, it will be able to so that liquid-phase chromatographic analysis is easy to operate, detection efficiency is improved,
It uses manpower and material resources sparingly, reduces use cost.
Summary of the invention
The technical problem to be solved by the invention is to provide a kind of binary channels on-line analysis pre-treatments and off-line analysis one
Liquid chromatograph and its application method, realization can complete off-line analysis and complete on-line analysis, and can be logical in different analyses
Different chromatography columns is used in road.The present invention can realize on a liquid chromatograph can either complete offline liquid phase color
Spectrum analysis, and on-line chromatograph analysis can be completed, realize the online switching of a variety of liquid chromatographic systems, and can be logical in different analyses
Different chromatography columns is used in road, is analyzed for substance of different nature.So that liquid-phase chromatographic analysis is easy to operate,
Detection efficiency is improved, is used manpower and material resources sparingly, use cost is reduced.
In order to solve above-mentioned technical purpose, the technical solution adopted by the present invention is that: a kind of binary channels on-line analysis pre-treatment
With off-line analysis one liquid chromatograph, including autosampler, two chromatogram pumps, three six-way valves, in-line purification column, two
Chromatography column, Photochemical derivatization device, UV detector and fluorescence detector, which is characterized in that first for system switching
Six interfaces of six-way valve respectively are a, b, c, d, e, f, six interfaces of the second six-way valve respectively be g, h, i, j,
K, l, six interfaces of third six-way valve respectively are m, n, o, p, q, r, autosampler respectively with the first chromatogram pump and
One six-way valve interface a is connected, and the first six-way valve interface f is connected with the second six-way valve interface g, the second chromatogram pump and third six-way valve
Interface n be connected, be connected with multiple liquid storage bottles on the second chromatogram pump, the interface p of the interface i of the second six-way valve and third six-way valve it
Between be connected with the first chromatography column, the interface j of the second six-way valve is connected with the interface o of third six-way valve, and the first six-way valve connects
The second chromatography column, the interface c and third six-way valve of the first six-way valve are connected between the mouth d and interface r of third six-way valve
Interface m be connected, be connected with Photochemical derivatization device, UV detector and fluorescence detector in turn on third six-way valve interface q, the
Quantitative loop is connected between one six-way valve interface b and interface e, the second six-way valve interface l is waste liquid interface, the second six-way valve interface
It is connected with in-line purification column between h and interface k, multiple liquid storage bottles are connected on the first chromatogram pump.
The application method of above-mentioned binary channels on-line analysis pre-treatment and off-line analysis one liquid chromatograph, including following step
It is rapid:
Step 1, when being used as off-line analysis, successively the following steps are included:
(1) sample introduction and system balancing process: autosampler injects a sample into system on the first chromatogram pump in Chu Ye bottle
Mobile phase enter the first six-way valve, sample in quantitative loop keep in, the second chromatogram pump will be in the Chu Ye bottle on the second chromatogram pump
Mobile phase bring third six-way valve into, into the first six-way valve, the second chromatography column, flow into third six-way valve and enter photochemistry
Derivatization device, UV detector and fluorescence detector, are balanced whole system；
(2) elution analysis process: the first six-way valve switches over, and the second chromatogram pump brings the mobile phase in liquid storage bottle into company
Quantitative loop on the first six-way valve is separated into the second chromatography column, and passes through Photochemical derivatization device, ultraviolet detection
Device and fluorescence detector are analyzed；
Step 2, when be used as online pre-treatment analysis when, successively the following steps are included:
(1) sample introduction and system balancing process: autosampler injects a sample into system on the first chromatogram pump in Chu Ye bottle
Mobile phase enter the first six-way valve, sample flows through quantitative loop and is brought into in-line purification column and is enriched with into the second six-way valve, miscellaneous
Matter and matrix are discharged into waste liquid with mobile phase；Second chromatogram pump brings the mobile phase in the second chromatogram pump liquid storage bottle into third six-way valve
The second six-way valve is transferred into the first chromatography column, Photochemical derivatization device, UV detector and fluorescence detector, to entire system
System is balanced；
(2) elution analysis process: the second six-way valve switches over, and the second chromatogram pump enters mobile phase through third six-way valve
The in-line purification column being connected on the second six-way valve, sample enters the first chromatography column and is separated in in-line purification column, and leads to
Photochemical derivatization device, UV detector and fluorescence detector is crossed to be analyzed；
(3) online activating and regenerating process: the second six-way valve switches over, and the first chromatogram pump will be in the first chromatogram pump liquid storage bottle
Mobile phase bring in-line purification column in the second six-way valve into, activating and regenerating is carried out to in-line purification column, subsequent first chromatogram pump will
Mobile phase in first chromatogram pump Chu Ye bottle is switched to sample introduction initial proportion than column, is balanced, is used for down to in-line purification column
The purification analysis of one sample.
Beneficial effects of the present invention: offline liquid-phase chromatographic analysis and online preceding place can be realized on a liquid chromatograph
The online switching for managing liquid-phase chromatographic analysis, can make of different nature according to the online analysis of shift channel of property of sample to be analysed
Substance realizes offline or online separation analysis in different chromatographic columns, solve Traditional liquid phase chromatograph can only do off-line analysis or
The shortcomings that person's in-line purification is analyzed, substantially increases the applicability and service efficiency of instrument, reduces the use cost of instrument.Together
When, in-line purification analysis system simplifies many and diverse operating process, saves the time of sample pre-treatments, and in-line purification column can be done
Activating and regenerating processing, is recycled, and to more environment-friendly, can also reduce use cost.
Detailed description of the invention
Fig. 1 is the offline inspection sample introduction of binary channels on-line analysis pre-treatment of the present invention and off-line analysis one liquid chromatograph
With the structural schematic diagram of system balancing.
Fig. 2 is binary channels on-line analysis pre-treatment of the present invention and the offline inspection elution of off-line analysis one liquid chromatograph
The structural schematic diagram of process.
Fig. 3 is that binary channels on-line analysis pre-treatment of the present invention and the online pre-treatment of off-line analysis one liquid chromatograph are examined
Survey the structural schematic diagram of sample introduction and system balancing and online activating and regenerating process.
Fig. 4 is that binary channels on-line analysis pre-treatment of the present invention and the online pre-treatment of off-line analysis one liquid chromatograph are examined
Survey the structural schematic diagram of elution process.
Fig. 5 is the chromatogram of aflatoxin in the cereal of the embodiment of the present invention.
Fig. 6 is the calibration graph of the aflatoxin G 2 of the embodiment of the present invention.
Fig. 7 is the calibration graph of the aflatoxin G 1 of the embodiment of the present invention.
Fig. 8 is the calibration graph of the aflatoxin B 2 of the embodiment of the present invention.
Fig. 9 is the calibration graph of the aflatoxin B1 of the embodiment of the present invention.
Figure 10 is the UV detector chromatogram of the online pre-treatment detection of bisphenol-A in the toner of the embodiment of the present invention.
Figure 11 is the fluorescence detector chromatogram of the online pre-treatment detection of bisphenol-A in the toner of the embodiment of the present invention.
Figure 12 is the UV detector calibration graph of the bisphenol-A of the embodiment of the present invention.
Figure 13 is the fluorescence detector calibration graph of the bisphenol-A of the embodiment of the present invention.
Specific embodiment
Technical solution of the present invention is described in further detail with reference to the accompanying drawings and detailed description:
As shown in Figs 1-4, binary channels on-line analysis pre-treatment of the invention and off-line analysis one liquid chromatograph, including
It is autosampler, two chromatogram pumps, three six-way valves, in-line purification column, two chromatography columns, Photochemical derivatization device, ultraviolet
Six interfaces of detector and fluorescence detector, the first six-way valve 3 for system switching respectively are a, b, c, d, e, f,
Six interfaces of the second six-way valve 5 respectively are g, h, i, j, k, l, six interfaces of third six-way valve 7 respectively be m,
N, o, p, q, r, autosampler 2 are connected with the first chromatogram pump 1 and 3 interface a of the first six-way valve respectively, 3 interface f of the first six-way valve
It is connected with 5 interface g of the second six-way valve, the second chromatogram pump 8 is connected with the interface n of third six-way valve 7, on the second chromatogram pump 8 respectively
It is connected with the 5th liquid storage bottle 17, the 6th liquid storage bottle 18, the 7th liquid storage bottle 19, the 8th liquid storage bottle 20, the interface i of the second six-way valve 5
The first chromatography column 21, the interface j and third six-way valve 7 of the second six-way valve 5 are connected between the interface p of third six-way valve 7
Interface o be connected, the second chromatography column 12 is connected between the interface d of the first six-way valve 3 and the interface r of third six-way valve 7,
The interface c of first six-way valve 3 is connected with the interface m of third six-way valve 7, is connected with photochemistry in turn on 7 interface q of third six-way valve
Derivatization device 9, UV detector 10 and fluorescence detector 11 are connected with quantitative loop 4 between first six-way valve, 3 interface b and interface e,
Second six-way valve, 5 interface l is waste liquid interface, is connected with in-line purification column 6 between the second six-way valve 5 interface h and interface k, first
The first liquid storage bottle 13, the second liquid storage bottle 14, third liquid storage bottle 15 and the 4th liquid storage bottle 16 are connected separately on chromatogram pump 1.
The application method of above-mentioned binary channels on-line analysis pre-treatment and off-line analysis one liquid chromatograph, including following step
It is rapid:
Step 1, when being used as off-line analysis, successively the following steps are included:
1 sample introduction and system balancing process: autosampler 2 injects a sample into system with the first storage on the first chromatogram pump 1
The mobile phase that liquid bottle the 13, the 2nd is stored up in the storage of liquid bottle 14 and the 3rd liquid bottle 15 enters the first six-way valve 3, and sample is temporary in quantitative loop 4
It deposits, the second chromatogram pump 8 is brought in the 5th liquid storage bottle 17, the 6th liquid storage bottle 18 and the 7th storage liquid bottle 19 mobile phase the into three six and led to
Valve 7 into the first six-way valve 3, the second chromatography column 12, flows into third six-way valve 7 into Photochemical derivatization device 9, ultraviolet inspection
Device 10 and fluorescence detector 11 are surveyed, whole system is balanced, as shown in Figure 1；
2 elution analysis processes: the first six-way valve 3 switches over, and the second chromatogram pump 8 is by the 5th liquid storage bottle 17, the 6th liquid storage
Mobile phase in bottle 18 and the 7th Chu Ye bottle 19 brings the quantitative loop 4 being connected on the first six-way valve 3 into, into the second chromatography column
12 are separated, and are analyzed by Photochemical derivatization device 9, UV detector 10 and fluorescence detector 11, are such as schemed, shown in 2；
Step 2, when be used as online pre-treatment analysis when, successively the following steps are included:
(1) sample introduction and system balancing process: autosampler 12 injects a sample into system with first on the first chromatogram pump 1
It stores up the mobile phase that liquid bottle 13, second is stored up in liquid bottle 14 and enters the first six-way valve 3, sample flows through quantitative loop 4 and enters the second six-way valve 5
It is brought into in-line purification column 6 and is enriched with, impurity and matrix are discharged into waste liquid with mobile phase；Second chromatogram pump 8 by the 5th liquid storage bottle 17,
Mobile phase in 6th liquid storage bottle 18, which brings third six-way valve 7 into and is transferred to the second six-way valve 5, enters the first chromatography column 21, photochemical
Derivatization device 9, UV detector 10 and fluorescence detector 11 are learned, whole system is balanced, as shown in Figure 3；
(2) elution analysis process: the second six-way valve 5 switches over, and the second chromatogram pump 8 is by mobile phase through third six-way valve 7
Into the in-line purification column 6 being connected on the second six-way valve 5, sample is carried out into the first chromatography column 21 in in-line purification column 6
Separation, and analyzed by Photochemical derivatization device 9, UV detector 10 and fluorescence detector 11, as shown in Figure 4；
(3) online activating and regenerating process: the second six-way valve 5 switches over, and the first chromatogram pump 1 is by the first storage liquid bottle 13 and the
Mobile phase in two liquid storage bottles 14 brings in-line purification column 6 in the second six-way valve 5 into, carries out activating and regenerating to in-line purification column, with
The mobile phase in the first storage liquid bottle 13 and the second liquid storage bottle 14 is switched to sample introduction initial proportion than column by the first chromatogram pump 1 afterwards, right
In-line purification column 6 is balanced, and the purification for next sample is analyzed, as shown in Figure 3.
Specifically, sample injector 2 is connected with the first chromatogram pump 1 and the first six-way valve 3, it is connected with quantitative loop 4 on the first six-way valve,
It is connected with the second chromatography column between first six-way valve 3 and third six-way valve, the second chromatogram pump 8 passes through third six-way valve 7 and the
One six-way valve 3 is connected, the first chromatogram pump 1 and the first liquid storage bottle 13, the second liquid storage bottle 14, third liquid storage bottle 15 and the 4th liquid storage bottle
16 are connected, and the second chromatogram pump is connected with the 5th liquid storage bottle 17, the 6th liquid storage bottle 18, the 7th liquid storage bottle 19 and the 8th liquid storage bottle 20,
Third six-way valve 7 is successively connect with Photochemical derivatization device 9, UV detector 10, fluorescence detector 11, constitutes offline liquid chromatogram
Analysis system；
Sample injector 2 is connected with the first chromatogram pump 1 and the first six-way valve 3, and the first six-way valve 3 is connected with quantitative loop 4, the first six-way valve
3 are connected with the second six-way valve, and in-line purification column 6 is connected on the second six-way valve, and the second chromatogram pump 8 is connected with third six-way valve 7, the
One chromatogram pump 1 is connected with the first liquid storage bottle 13, the second liquid storage bottle 14, third liquid storage bottle 15 and the 4th liquid storage bottle 16, the second chromatography
Pump is connected with the 5th liquid storage bottle 17, the 6th liquid storage bottle 18, the 7th liquid storage bottle 19 and the 8th liquid storage bottle 20, connects on the second six-way valve 5
Coloured first spectrum splitter 9, chromatography column 9 is connected with third six-way valve 7, third six-way valve 7 successively with Photochemical derivatization device
9, UV detector 10, fluorescence detector 11 connect into in-line purification pretreatment liquid chromatographic system.
First liquid storage bottle 13 is used for armoring alcohol for filling water, the second liquid storage bottle 14, and third liquid storage bottle 15 is used to fill acetonitrile, the
Four liquid storage bottles 16 are spare.
5th liquid storage bottle 17 is used for armoring alcohol for filling water, the 6th liquid storage bottle 18, and the 7th liquid storage bottle 19 fills acetonitrile, the 8th storage
Liquid bottle 20 is spare.
It is embodied as follows:
When this liquid chromatographic system does aflatoxin in off-line analysis cereal, its step are as follows:
(1) sample introduction and system balancing process: weighing 5g sample in 50mL centrifuge tube, and methanol-water solution (70+ is added
30) it, is vortexed and mixes, vibrate 20min, 6000r/min is centrifuged 10min, takes 4.0mL supernatant, and 46mL1%TritonX- is added
100 PBS is mixed, and crosses immune affinity column, the elution of 2 × 10mL water, and 2 × 1mL methanol elutes, and 50 DEG C of nitrogen are blown to close dry, initial flow
It is dynamic mutually to redissolve to 1.0mL, it is vortexed and mixes 30s, 0.22um membrane filtration, HPLC measurement.Autosampler 2 injects 50 μ L samples
System is in the water in the first storage liquid bottle 13 in the first chromatogram pump 1, the methanol and the 3rd storage liquid bottle 15 in the 2nd storage liquid bottle 14
Acetonitrile (water+methanol+acetonitrile=68+16+16) enter the first six-way valve 3, injecting 50 μ L altogether, (speed of 0.25mL/min is injected
0.2min), sample in quantitative loop 4 keep in, the second chromatogram pump 8 with the speed of 1.0mL/min by the 5th liquid storage bottle 17 water,
The acetonitrile (water+methanol+acetonitrile=68+16+16) in methanol and the 7th storage liquid bottle 19 in 6th liquid storage bottle 18 brings the three or six into
Port valve 7, into the first six-way valve 3,12 (model: Utimate XB-C18,4.6*150mm, 5 μm, column of the second chromatography column
Temperature: 30 DEG C), flow into third six-way valve 7 and enter Photochemical derivatization device 9, UV detector 10 and fluorescence detector 11, to entire system
System is balanced；
(2) elution analysis process: the first six-way valve 3 switches over, and the second chromatogram pump 8 is with the speed of 1.0mL/min by
Water in five liquid storage bottles 17, the methanol in the 6th liquid storage bottle 18 and the acetonitrile (water+methanol+acetonitrile=68+ in the 7th Chu Ye bottle 19
The quantitative loop 4 being connected on the first six-way valve 3 16+16) is brought into, into 12 (model: Utimate XB- of the second chromatography column
C18,4.6*150mm, 5 μm, column temperature: 30 DEG C) it is separated, and pass through Photochemical derivatization device 9, UV detector 10 and fluorescence inspection
Device 11 (excitation wavelength 360nm, launch wavelength 440nm) is surveyed to be analyzed.Weaker, this application example of the ultraviolet response of aflatoxin
Using only fluorescence detector, UV detector is spare when being other analyses.
(4) aflatoxin G 2 and aspergillus flavus poison the drafting of standard curve: are prepared with water+methanol+acetonitrile=68+16+16
Plain B2 concentration is 0.0577ng/mL, 0.115ng/mL, 0.231ng/mL, 0.462ng/mL, 0.923ng/mL, aflatoxin
G1 and aflatoxin B1 concentration are 0.192ng/mL, 0.385ng/mL, 0.769ng/mL, 1.538ng/mL, 3.077ng/mL
Calibration curve solution, linear regression is carried out with mass concentration (X) and peak area (Y).Obtain 2 calibration curve of aflatoxin G
Linear equation be Y=1673.6174X+55.0985, linearly dependent coefficient R2=0.9996, aflatoxin G 1 calibration curve
Linear equation be Y=815.4444X+38.5734, linearly dependent coefficient R2=0.9994,2 calibration curve of aflatoxin B
Linear equation be Y=3641.8089X+30.7431, linearly dependent coefficient R2=0.9991, aflatoxin B1 calibration curve
Linear equation be Y=1699.8684X+102.0725, linearly dependent coefficient R2=0.9992.Aflatoxin chromatogram is shown in
Fig. 5,2 standard curve of aflatoxin G are shown in that Fig. 6, aflatoxin G 1 standard curve are shown in Fig. 7,2 standard curve of aflatoxin B
See that Fig. 8, aflatoxin B1 standard curve are shown in Fig. 9.
When this liquid chromatographic system does bisphenol-A in online pre-treatment analysis toner, its step are as follows:
(1) sample introduction and system balancing process: weighing 1g sample in 50mL centrifuge tube, and 5g sodium chloride solution, shaking is added
It mixes, 10min is extracted in addition 10mL methanol, concussion, and 4000r/min is centrifuged 3min, after supernatant crosses 0.45um filter membrane, for dividing
Analysis.Autosampler 2 is injected a sample into system and is stored up in liquid bottle 13 with the speed of 0.8mL/min by the first with the first chromatogram pump 1
Methanol (water+methanol=70+30) in water, the second storage liquid bottle 14 enters the first six-way valve 3, and sample flows through quantitative loop 4 and enters the
Two six-way valves 5 are brought into in-line purification column 6 (model: 3 μm of Acclaim PolarAdvantage II, 4.6*50mm) rich
Collection, impurity and matrix are discharged into waste liquid with mobile phase；Second chromatogram pump 8 with 0.8mL/min by the 5th liquid storage bottle 17 water, the 6th
Methanol in liquid storage bottle 18 bring into third six-way valve 7 be transferred to the second six-way valve 5 enter the first chromatography column 21 (model:
Waters SYMMETRY 4.6mm*150mm, 5um, column temperature: 30 DEG C), Photochemical derivatization device 9, UV detector 10 and fluorescence inspection
Device 11 is surveyed, whole system is balanced.
(2) elution analysis process: the second six-way valve 5 switches over, and the second chromatogram pump 8 is with 0.8mL/min by the 5th liquid storage
Water in bottle 17, the methanol in the 6th liquid storage bottle 18 enter the in-line purification column being connected on the second six-way valve 5 through third six-way valve 7
6 (models: 3 μm of Acclaim PolarAdvantage II, 4.6*50mm), sample enters the first chromatography in in-line purification column 6
Splitter 21 (model: Waters SYMMETRY 4.6mm*150mm, 5um, column temperature: 30 DEG C) is separated, and passes through photochemistry
Derivatization device 9, UV detector 10 (wavelength 280nm) and fluorescence detector 11 (excitation wavelength 230nm, launch wavelength 313nm) into
Row analysis.This application example does not use Photochemical derivatization device.
(3) washing of in-line purification column and activation process: the second six-way valve switches over 5, and the first chromatogram pump 1 is with 0.8mL/
Min by the first six-way valve 3, brings the methanol in the water and the 6th liquid storage bottle 14 in the 5th Chu Ye bottle 13 into second six-way valve 5
In in-line purification column 6 (model: 3 μm of Acclaim PolarAdvantage II, 4.6*50mm), to 6 (type of in-line purification column
Number: 3 μm of Acclaim PolarAdvantage II, 4.6*50mm) activating and regenerating is carried out, subsequent first chromatogram pump 1 is by first
Methanol in water and the second liquid storage bottle 14 in Chu Ye bottle 13 is switched to sample introduction initial proportion than column, carries out to in-line purification column 6
Balance, the purification for next sample are analyzed
1 first liquid storage bottle 13 of (4) first chromatogram pump, 14 time gradient program of the second liquid storage bottle see the table below:
| Time min | Liquid storage bottle 13 (%) | Liquid storage bottle 14 (%) |
| 0 | 70 | 30 |
| 3 | 70 | 30 |
| 12.5 | 10 | 90 |
| 13.0 | 70 | 30 |
(5) second 1 the 5th liquid storage bottles 17 of spectrum pump, 18 time gradient program of the 6th liquid storage bottle see the table below:
| Time min | Liquid storage bottle 17 (%) | Liquid storage bottle 18 (%) |
| 0 | 30 | 70 |
| 9 | 30 | 30 |
| 12.5 | 10 | 90 |
| 13.0 | 30 | 70 |
(6) 0mg/L, 0.03mg/L, 0.06mg/L, 0.15mg/L, 0.30mg/ the drafting of standard curve: are prepared with methanol
L,0.75mg/L.Calibration curve solution, linear regression is carried out with mass concentration (X) and peak area (Y).Obtain UV detector
The linear equation of calibration curve is Y=2.4649X-0.0202, linearly dependent coefficient R2=0.9999, fluorescence detector calibration song
The linear equation of line is Y=2606104.9071X+5146.4622, linearly dependent coefficient R2=0.9999.Bisphenol-A ultraviolet detection
Device chromatogram is shown in that Figure 10, bisphenol-A fluorescence detector chromatogram are shown in that Figure 11, bisphenol-A UV detector standard curve are shown in Figure 12, bis-phenol
A UV detector standard curve is shown in Figure 13.
Embodiment described above is merely to illustrate technical idea and feature of the invention, in the art its object is to make
Technical staff it will be appreciated that the contents of the present invention and implement accordingly, patent model of the invention only cannot be limited with the present embodiment
It encloses, i.e., same changes or modifications made by all disclosed spirit are still fallen in the scope of the patents of the invention.
Claims (2)
1. a kind of binary channels on-line analysis pre-treatment and off-line analysis one liquid chromatograph, including autosampler, two colors
Compose pump, three six-way valves, in-line purification column, two chromatography columns, Photochemical derivatization device, UV detector and fluorescence detection
Device, which is characterized in that six interfaces of the first six-way valve (3) for system switching respectively are a, b, c, d, e, f, and second
Six interfaces of six-way valve (5) respectively are g, h, i, j, k, l, six interfaces of third six-way valve (7) respectively be m,
N, o, p, q, r, autosampler (2) are connected with the first chromatogram pump (1) and the first six-way valve (3) interface a respectively, the first six-way valve
(3) interface f is connected with the second six-way valve (5) interface g, and the second chromatogram pump (8) is connected with the interface n of third six-way valve (7), and second
It is connected with multiple liquid storage bottles on chromatogram pump (8), is connected between the interface i of the second six-way valve (5) and the interface p of third six-way valve (7)
The interface j of first chromatography column (21), the second six-way valve (5) is connected with the interface o of third six-way valve (7), the first six-way valve
(3) it is connected with the second chromatography column (12) between the interface r of interface d and third six-way valve (7), the first six-way valve (3)
Interface c is connected with the interface m of third six-way valve (7), is connected with Photochemical derivatization device in turn on third six-way valve (7) interface q
(9), UV detector (10) and fluorescence detector (11), are connected with quantitative loop between the first six-way valve (3) interface b and interface e
(4), the second six-way valve (5) interface l is waste liquid interface, is connected with in-line purification between the second six-way valve (5) interface h and interface k
Column (6), the first chromatogram pump are connected with multiple liquid storage bottles on (1).
2. the application method of binary channels on-line analysis pre-treatment as described in claim 1 and off-line analysis one liquid chromatograph,
Characterized by comprising the following steps:
Step 1, when being used as off-line analysis, successively the following steps are included:
(1) sample introduction and system balancing process: autosampler (2) injects a sample into system with Chu Ye bottle on the first chromatogram pump (1)
In mobile phase enter the first six-way valve (3), sample in quantitative loop (4) keep in, the second chromatogram pump (8) will be on the second chromatogram pump
Chu Ye bottle in mobile phase bring third six-way valve (7) into, into the first six-way valve (3), the second chromatography column (12), flow into
Third six-way valve (7) enters Photochemical derivatization device (9), UV detector (10) and fluorescence detector (11), to whole system into
Row balance；
(2) elution analysis process: the first six-way valve (3) switches over, and the second chromatogram pump (8) brings the mobile phase in liquid storage bottle into
The quantitative loop (4) being connected on the first six-way valve (3), is separated, and spread out by photochemistry into the second chromatography column (12)
Raw device (9), UV detector (10) and fluorescence detector (11) are analyzed；
Step 2, when be used as online pre-treatment analysis when, successively the following steps are included:
(1) sample introduction and system balancing process: autosampler (12) injects a sample into system with Chu Ye bottle on the first chromatogram pump (1)
In mobile phase enter the first six-way valve (3), sample flows through quantitative loop (4) and is brought into in-line purification into the second six-way valve (5)
Enrichment in column (6), impurity and matrix are discharged into waste liquid with mobile phase；Second chromatogram pump (8) is by the stream in the second chromatogram pump liquid storage bottle
Dynamic phase bring into third six-way valve (7) be transferred to the second six-way valve (5) into the first chromatography column (21), Photochemical derivatization device (9),
UV detector (10) and fluorescence detector (11), are balanced whole system；
(2) elution analysis process: the second six-way valve (5) switches over, and the second chromatogram pump (8) is by mobile phase through third six-way valve
(7) enter the in-line purification column (6) being connected on the second six-way valve (5), sample enters the first chromatographic isolation in in-line purification column (6)
Column (21) is separated, and is analyzed by Photochemical derivatization device (9), UV detector (10) and fluorescence detector (11)；
(3) online activating and regenerating process: the second six-way valve (5) switches over, and the first chromatogram pump (1) is by the first chromatogram pump liquid storage bottle
In mobile phase bring in-line purification column (6) in the second six-way valve (5) into, to in-line purification column carry out activating and regenerating, subsequent first
Chromatogram pump (1) by the first chromatogram pump Chu Ye bottle mobile phase than column is switched to sample introduction initial proportion, to in-line purification column (6) into
Row balance, the purification for next sample are analyzed.
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| WO2021103658A1 (en) * | 2019-11-25 | 2021-06-03 | 清华大学 | Automatic sample injection and analysis device for multiple samples |
| CN113295810A (en) * | 2021-05-26 | 2021-08-24 | 天津海关动植物与食品检测中心 | Online analysis method for overlapped sample injection of chromatographic system |
| CN113391010A (en) * | 2021-07-07 | 2021-09-14 | 安图实验仪器(郑州)有限公司 | Sample introduction method of clinical automatic analysis instrument |
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| CN106645502A (en) * | 2017-02-27 | 2017-05-10 | 天津出入境检验检疫局动植物与食品检测中心 | Ion chromatograph based on multi-valve switching and for offline and online analysis and detection method of ion chromatograph |
| CN106872596A (en) * | 2017-02-27 | 2017-06-20 | 天津出入境检验检疫局动植物与食品检测中心 | A kind of binary channels based on many Vavle switchings offline and on-line analysis ion chromatograph and its detection method |
| CN209858493U (en) * | 2019-04-25 | 2019-12-27 | 天津出入境检验检疫局动植物与食品检测中心 | Dual-channel online analysis pretreatment and offline analysis integrated liquid chromatograph |
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| CN106645502A (en) * | 2017-02-27 | 2017-05-10 | 天津出入境检验检疫局动植物与食品检测中心 | Ion chromatograph based on multi-valve switching and for offline and online analysis and detection method of ion chromatograph |
| CN106872596A (en) * | 2017-02-27 | 2017-06-20 | 天津出入境检验检疫局动植物与食品检测中心 | A kind of binary channels based on many Vavle switchings offline and on-line analysis ion chromatograph and its detection method |
| CN209858493U (en) * | 2019-04-25 | 2019-12-27 | 天津出入境检验检疫局动植物与食品检测中心 | Dual-channel online analysis pretreatment and offline analysis integrated liquid chromatograph |
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| WO2021103658A1 (en) * | 2019-11-25 | 2021-06-03 | 清华大学 | Automatic sample injection and analysis device for multiple samples |
| CN113295810A (en) * | 2021-05-26 | 2021-08-24 | 天津海关动植物与食品检测中心 | Online analysis method for overlapped sample injection of chromatographic system |
| CN113391010A (en) * | 2021-07-07 | 2021-09-14 | 安图实验仪器(郑州)有限公司 | Sample introduction method of clinical automatic analysis instrument |
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