CN101038274A - Pressurized capillary electrochromatography chemiluminescence detecting method - Google Patents
Pressurized capillary electrochromatography chemiluminescence detecting method Download PDFInfo
- Publication number
- CN101038274A CN101038274A CN 200710008810 CN200710008810A CN101038274A CN 101038274 A CN101038274 A CN 101038274A CN 200710008810 CN200710008810 CN 200710008810 CN 200710008810 A CN200710008810 A CN 200710008810A CN 101038274 A CN101038274 A CN 101038274A
- Authority
- CN
- China
- Prior art keywords
- chemiluminescence
- capillary
- detecting method
- pressurized
- electrochromatographchemiluminescence
- 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.)
- Pending
Links
Images
Landscapes
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Abstract
The present invention provides a pressurized capillary electrochromatography chemiluminescence detection device comprising a liquid storage tank of mobile phase, a high pressure pump used for carrying the mobile phase and providing liquid to a separation capillary electroosmotic chromatography connected to a detection cell through a microflow controller in which a sample can be input, a high voltage power supply which both ends are applied respectively to a liquid intake terminal of the separation capillary electroosmotic chromatography and a detection cell, and a photomultiplier used for collecting optic signals of sample to be detected, wherein said photomultiplier is set below the detection cell and its output terminal is connected to a computer through a signal amplifier. By combining a chemiluminescence analysis method and a pressurized capillary electrochromatography technology, the present invention capable of operated easily and having detection sensitivity is suitable for applications in fields of food safety, environmental monitoring, clinical medicine, and the like, and is easily to extend.
Description
Technical field
The present invention relates to a kind of analytical instrument that is used for the separation detection complex sample, be specifically related to a kind of pressurized capillary electrochromatographchemiluminescence chemiluminescence detecting method.
Background technology
Pressurization capillary electric chromatogram (Pressurized Capillary Electrochromatography, abbreviation pCEC) high efficiency of Capillary Electrophoresis (CE) and the high selectivity of high performance liquid chromatography (HPLC) have been merged, be a kind of novel Micro-Column Separation analytical technology (Tsuda, T.Anal.Chem.1988,60,1677-1680).Have the separation efficiency height, analysis time is short, sample size is little, reagent consumption less, superior function such as easily be automated.In pCEC used, because sampling volume little (nL level), characteristics such as component concentration to be measured is low in the trickle and sample of caliber were had relatively high expectations to the sensitivity of detecting device, and lacking high-sensitive detecting device has become and limit the principal element that this technology is used.Chemiluminescence (Chemiluminescence, CL) analytic approach need not add light source, thereby background is low, can obtain higher highly sensitive, the maximum characteristics of this type of detecting device be instrument and equipment simple, easy to operate, be easy to and various isolation technics couplings.PCEC and CL detection technique being combined, can have both the high sensitivity of CL and the characteristics of pCEC high efficiency simultaneously, is method for separating and analyzing comparatively desirable in the microanalysis field, does not have relevant technology report at present.
Summary of the invention
The objective of the invention is at the problems referred to above, chemiluminometry and pressurization capillary electric chromatogram isolation technics are combined, a kind of pressurized capillary electrochromatographchemiluminescence chemiluminescence detecting method is provided.
Pressurized capillary electrochromatographchemiluminescence chemiluminescence detecting method of the present invention, the liquid storage container (1) that comprises moving phase, in order to transmit the high-pressure pump (2) of moving phase, high-voltage power supply (16), the microfluidic control instrument of high-pressure pump through importing sample is to separation capillary electric chromatographic column (9) feed flow that inserts detection cell (10), the two ends of high-voltage power supply put on the liquid feeding end and the detection cell of described separation capillary electric chromatographic column (9) respectively, described detection cell below is placed with the photomultiplier (12) that is used to gather the determinand light signal, and the output terminal of described photomultiplier (12) is connected with computing machine (14) through signal amplifier (13).
Remarkable advantage of the present invention is:
One: chemiluminometry and pressurization capillary electric chromatogram isolation technics are combined, can have both the high sensitivity of chemiluminometry and the characteristics of pressurization capillary electric chromatogram isolation technics high efficiency simultaneously, be method for separating and analyzing comparatively desirable in the microanalysis field.The present invention is a detected object with the luminescent substance luminol of classics, and by debugging and optimization pCEC-CL device, the detection that records luminol is limited to 2.0 * 10
-10Mol/L, and utilize this device successfully to realize separation and detection to biological samples such as amino acid having obtained satisfactory experimental results.
Two: apparatus of the present invention processing ease, the detection sensitivity height is suitable for the application in fields such as food security, environment measuring, clinical medicine, is easy to promote.
Description of drawings
Fig. 1 is the apparatus structure general illustration that the present invention designs, the liquid storage container of 1-moving phase wherein, 2-high-pressure pump, the 3-six-way injection valve, 4-sample introduction end, 5-waste liquid end, 6-four-way shunt, 7-prevents electronic box, the 8-backpressure valve, 9-separation capillary electric chromatographic column, 10-detection cell, 11-kapillary, the 12-photomultiplier, 13-signal amplifier, 14-computing machine, the 15-waste liquid pool, the 16-high-voltage power supply.
Fig. 2 is the electrochromatogram that pressurized capillary electrochromatographchemiluminescence chemiluminescence detecting method separates and detect luminol.
Fig. 3 is that pressurized capillary electrochromatographchemiluminescence chemiluminescence detecting method separates and detection threonine and tyrosine electrochromatogram.
Embodiment
Microfluidic control instrument of the present invention comprises six-way injection valve (3), four-way diverting valve (6) and backpressure valve (8), the delivery outlet of described six-way injection valve is connected to the liquid feeding end of separation capillary electric chromatographic column (9) through the four-way diverting valve, and the another port of four-way diverting valve is connected with the input end of the backpressure valve that leads to waste liquid pool.
Capillary electric chromatographic column (9) can be a packed column, any one in integral post or the open tubular column.
The chemiluminescence detection pond can be the styletable formula, a kind of in post or post rear sleeve formula chemiluminescence detection pond.
Four-way diverting valve, backpressure valve, waste liquid pool, high-voltage power supply, detection cell, photomultiplier and signal amplifier are positioned in the anti-electronic box (7).
The operating process of this device is as follows:
Stock moving phase in liquid storage container (1) under the promotion of high-pressure pump (2), be delivered to six-way injection valve (3), testing sample is through the injection of (4) position, switch six-way injection valve (3) immediately, unnecessary testing sample is discharged from (5) position, quantitative testing sample (2 μ L) is then brought four-way shunt (6) under the promotion of moving phase, keep constant post to press by regulating backpressure valve (8), the testing sample of about at last 1/250 volume enters capillary electric chromatographic column (9), when high-voltage power supply (16) when applying high pressure, testing sample is under the double drive of electroosmotic flow and pressure current, in capillary electric chromatographic column (9), effectively separated, and flow out the post end successively, fully react with luminescence reagent in the detection cell (10) and produce luminous signal, gather through photomultiplier (12), transfer to that signal amplifier (13) is accepted and output detection signal to computing machine (14), reaction back solution is discharged by kapillary (11), and earth terminal is placed on and constitutes whole loop in the waste liquid pool (15).
Utilizing pressurized capillary electrochromatographchemiluminescence chemiluminescence detecting method, is detected object with the luminescent substance luminol of classics, examines or check this system performance, the results are shown in Figure 2.The result shows that this instrument can satisfy requirement of experiment, highly sensitive, favorable reproducibility.Experiment condition is: stationary phase is 3 μ m ODS-C18 posts; Moving phase is acetonitrile: phosphate=30: 70 (%v/v; PH8.0; 5mmol/L; Contain Co
2+: 1.0 * 10
-4Mol/L); Flow velocity is 0.01ml/min; Luminescence reagent is H behind the post
2O
2(concentration is 100mmol/L) transfers to pH12.30 with NaOH; The luminescence reagent flow velocity is 1.3 μ L/min behind the post; Separation voltage is-4kv; Testing result: the luminol range of linearity is 2.0 * 10
-9～2.0 * 10
-6Mol/L (r=0.9960); Detection is limited to 2.0 * 10
-10Mol/L.
Utilize pressurized capillary electrochromatographchemiluminescence chemiluminescence detecting method, measure threonine and tyrosine, the results are shown in Figure 3.Experiment condition is: polymerized (IBMA-EDMA-AMPS) integral post; Moving phase is methyl alcohol: phosphate=20: 80 (%v/v; PH8.0; 5mmol/L; Contain luminol:1.0 * 10
-4Mol/L; Cu
2+: 1.0 * 10
-5Mol/L); Flow velocity is 0.01ml/min; Luminescence reagent is H behind the post
2O
2(concentration is 100mmol/L) transfers to pH12.30 with NaOH; The luminescence reagent flow velocity is 1.3 μ L/min behind the post; Sample: 1-threonine (1.68 * 10
-4Mol/L); 2-tyrosine (1.2 * 10
-4Mol/L).
Claims (5)
1. pressurized capillary electrochromatographchemiluminescence chemiluminescence detecting method, the liquid storage container (1) that comprises moving phase, in order to transmit the high-pressure pump (2) of moving phase, high-voltage power supply (16), it is characterized in that: the microfluidic control instrument of described high-pressure pump through importing sample is to separation capillary electric chromatographic column (9) feed flow that inserts detection cell (10), the two ends of high-voltage power supply put on the liquid feeding end and the detection cell (10) of described separation capillary electric chromatographic column (9) respectively, described detection cell below is placed with the photomultiplier (12) that is used to gather the determinand light signal, and the output terminal of described photomultiplier (12) is connected with computing machine (14) through signal amplifier (13).
2. pressurized capillary electrochromatographchemiluminescence chemiluminescence detecting method according to claim 1, it is characterized in that: described microfluidic control instrument comprises six-way injection valve (3), four-way diverting valve (6) and backpressure valve (8), the delivery outlet of described six-way injection valve is connected to the liquid feeding end of separation capillary electric chromatographic column (9) through the four-way diverting valve, and the another port of four-way diverting valve is connected with the input end of the backpressure valve that leads to waste liquid pool.
3. pressurized capillary electrochromatographchemiluminescence chemiluminescence detecting method according to claim 1 and 2 is characterized in that: described separation capillary electric chromatographic column is a packed column, a kind of in integral post and the open tubular column.
4. pressurized capillary electrochromatographchemiluminescence chemiluminescence detecting method according to claim 1 and 2 is characterized in that: described chemiluminescence detection pond is the styletable formula, a kind of in post or post rear sleeve formula chemiluminescence detection pond.
5. pressurized capillary electrochromatographchemiluminescence chemiluminescence detecting method according to claim 1 and 2 is characterized in that: described four-way diverting valve, backpressure valve, waste liquid pool, high-voltage power supply, detection cell, photomultiplier and signal amplifier are positioned in the anti-electronic box (7).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200710008810 CN101038274A (en) | 2007-04-10 | 2007-04-10 | Pressurized capillary electrochromatography chemiluminescence detecting method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200710008810 CN101038274A (en) | 2007-04-10 | 2007-04-10 | Pressurized capillary electrochromatography chemiluminescence detecting method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101038274A true CN101038274A (en) | 2007-09-19 |
Family
ID=38889317
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200710008810 Pending CN101038274A (en) | 2007-04-10 | 2007-04-10 | Pressurized capillary electrochromatography chemiluminescence detecting method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101038274A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101561425B (en) * | 2009-06-02 | 2012-03-14 | 福州大学 | Electrochemiluminescence detection device of pressurized-capillary electrochromatography |
| CN110672761A (en) * | 2019-09-26 | 2020-01-10 | 上海通微分析技术有限公司 | A pressurization capillary electrochromatography device for quantitative and qualitative analysis |
-
2007
- 2007-04-10 CN CN 200710008810 patent/CN101038274A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101561425B (en) * | 2009-06-02 | 2012-03-14 | 福州大学 | Electrochemiluminescence detection device of pressurized-capillary electrochromatography |
| CN110672761A (en) * | 2019-09-26 | 2020-01-10 | 上海通微分析技术有限公司 | A pressurization capillary electrochromatography device for quantitative and qualitative analysis |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101871856B (en) | Solid phase extraction and concentration system | |
| US8343774B2 (en) | Chromatography-based monitoring and control of multiple process streams | |
| US9354209B2 (en) | Miniature HPLC device | |
| CN101782587B (en) | Instrument system suitable for synchronous on-line detection of adsorption spectrum and fluorescence spectrum | |
| CN107589190B (en) | Large-volume sample introduction-double solid phase extraction-high performance liquid chromatography online combined equipment | |
| CN1740779A (en) | Analyzing system and analysing method based on integrated micro-flow control chip | |
| CN101561425B (en) | Electrochemiluminescence detection device of pressurized-capillary electrochromatography | |
| Hsu et al. | Multifunctional microchip-based distillation apparatus I-Steam distillation for formaldehyde detection | |
| US7977114B2 (en) | Method and apparatus for monitoring processes using a single detector | |
| CN1828290A (en) | Method for analyzing trace organic substance in water using on-line combined solid phase extraction and liquid chromatography | |
| CN101038271A (en) | Pressurized capillary electrochromatography electrochemistry detecting method | |
| CN101038274A (en) | Pressurized capillary electrochromatography chemiluminescence detecting method | |
| CN101038255A (en) | Capillary electrophoresis chemiluminescence detector of acridiniumester, acridine sulfonamide and marker thereof, and method thereof | |
| CN102565255B (en) | PCEC (Pressurized Capillary Electrochromatography) ELSD (Evaporative Light-Scattering Detector) | |
| CN102116762A (en) | Multi-point sampling sample injector applied to ion mobility spectrometry | |
| Duong et al. | Inexpensive and versatile measurement tools using purpose-made capillary electrophoresis devices coupled with contactless conductivity detection: A view from the case study in Vietnam | |
| CN103884789B (en) | Method for rapidly determining polysaccharide peptide in lucid ganoderma product | |
| CN102680590A (en) | Two-dimensional liquid chromatogram (LC)-tandem mass spectrometry method for detecting four amine substances in water | |
| Llorent-Martínez et al. | Sequential injection multi-optosensor based on a dual-luminescence system using two sensing zones: application to multivitamin determination | |
| CN214252161U (en) | Liquid chromatogram-flow stopping system and circular dichroism spectrometer combined device | |
| CN106979985B (en) | Liquid chromatogram atomic spectrum combined system | |
| Cerdà et al. | Continuous-Flow Extraction | |
| Kissinger et al. | High performance liquid chromatography experiments for undergraduate laboratories | |
| CN202083681U (en) | Pressurized capillary electrochromatography evaporative light scattering detector | |
| CN215641050U (en) | Two-dimensional liquid chromatography detection device based on LSPR technology |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Open date: 20070919 |