CN104007219A - Liquid chromatography and enzyme electrode combining device and application thereof - Google Patents
Liquid chromatography and enzyme electrode combining device and application thereof Download PDFInfo
- Publication number
- CN104007219A CN104007219A CN201410248217.4A CN201410248217A CN104007219A CN 104007219 A CN104007219 A CN 104007219A CN 201410248217 A CN201410248217 A CN 201410248217A CN 104007219 A CN104007219 A CN 104007219A
- Authority
- CN
- China
- Prior art keywords
- enzyme electrode
- liquid chromatography
- organic solvent
- chromatographic
- detecting device
- 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
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a liquid chromatography and enzyme electrode combining device. The device comprises a chromatographic separation unit, a detector, an organic-solvent removing unit, an activity evaluation unit and a computer, wherein the chromatographic separation unit comprises a mobile-phase storage tank, a chromatographic pump, a sampling valve and a liquid chromatography column which are connected in sequence by pipelines; the liquid chromatography column is connected with the detector by the pipeline; the organic-solvent removing unit is used for removing an organic solvent in a sample, and the front end of the organic-solvent removing unit is connected with the detector; the activity evaluation unit comprises a mixer, a buffer-solution storage tank and an enzyme electrode; the mixer is connected with the rear end of the organic-solvent removing unit; the buffer-solution storage tank is connected to the mixer which is connected to the enzyme electrode; the computer is respectively and electrically connected with the detector and the enzyme electrode. The liquid chromatography and enzyme electrode combining device has the advantages that the biological activity of all components in the sample can be provided and analyzed in an online and real-time manner, the analysis on the components in the sample to be detected can be realized and the corresponding biological activity of all the components can be evaluated in an online manner.
Description
Technical field
The present invention relates to one and belong to drug world, relate in particular to a kind of liquid chromatography-enzyme electrode combined apparatus and traditional Chinese medicine ingredients-active on-line analysis application.
Background technology
Along with the expansion of Chinese Medicine Industry scale and the development of traditional Chinese medicine research achievement, Analysis of Chinese Traditional Medicine technology has obtained very big progress, and the object of Analysis of Chinese Traditional Medicine more concentrates on for the key scientific problems in traditional Chinese medicine Study on Modernization; Analysis of Chinese Traditional Medicine need to be integrated various new methods, new technology to make it to become the new system that solves key scientific problems simultaneously, and is no longer confined to analyze the material composition of Chinese medicine, and what more focus on solving is the composition of material and the correlativity of function thereof.The development of total system biology (GlobalSystems Biology), system disclose that the variation of chemical matterome and biosystem reply time null response correlativity; And square medicine fingerprint-medicine based on metabolism group is for the research imagination of the Chinese medicine compound prescription metabolism of marker-drug effect target three dimensional pattern, has all embodied and has improved existing Analysis of Chinese Traditional Medicine new technology, the development trend of confluence analysis method application.
Traditional Chinese medicine ingredients analytical technology is on the basis of liquid chromatography in the past, gas chromatography, thin-layer chromatography, the technology such as gas phase-mass spectrometry, liquid phase-mass spectrometry and even the coupling of chromatogram-nuclear-magnetism are developed, when carrying out compartment analysis, centering prodrug complex composition provides abundant structural information, known in Chinese medicine or the qualitative and quantitative analysis of principal component are not realized, greatly advance traditional Chinese medicine ingredients chemical research, but existing analytical technology is all taking chemical constitution study as object, cannot react online or the biologically active of evaluative component.Therefore, select the zymoprotein molecule relevant to herbal function, make fixed enzyme membrane by physical chemistry crosslinked action immobilization, then be combined with electrochemical sensor and form enzyme electrode (enzyme electrode), utilize selectivity, stability and electrochemical analysis quick, sensitive of enzyme reaction, the biologically active of each composition in analytic sample is provided online, in real time.Enzyme electrode is combined liquid phase analysis, can break through the disappearance of composition activated information in existing analytical technology, provides thinking and technical scheme for setting up new medicine quality evaluated pattern.
Electrode analysis method (comprising enzyme electrode), as a kind of Electrochemical Detection means, has successfully realized the on-line coupling with chromatography of ions.But due to the existence of organic solvent in liquid chromatogram mobile phase, limit the coupling of enzyme electrode and liquid chromatography.If organic solvent removal device is connected between liquid chromatography and enzyme electrode, organic solvent in mobile phase is removed, can realize the coupling of liquid chromatography and enzyme electrode.
In stratographic analysis, enrichment method is the conventional pre-treatment means of sample, before sample enters chromatographic column separation, carries out enrichment method.Also have enrichment method after post, such as Liquid Chromatography-Mass Spectrometry (LC-MS) and liquid chromatography-evaporative light-scattering method (LC-ELSD), but they are all mass flow rate sensitive detector but not concentration detector.If connect a removal of solvents device after normal concentration type detecting device, employing has poriness and hydrophobic voided polytetrafluoroethylene film and hollow-fibre membrane as permeable membrane, the infiltration of permission organic solvent, can realize removal of solvents, and reach the concentrated object of sample component, for further activity rating provides basis.Organic solvent removal device design: when flow communication is crossed porous permeable membrane pipe, cross the dry gas (such as air or nitrogen) having heated at tube wall circumferential flux, organic solvent is heated volatilization for gaseous state, sees through permeable membrane and removes.
Traditional Chinese medicine ingredients composition is complicated, and conventional analytical technology has been brought into play Chinese medicine effect in the qualitative and quantitative analysis of Chinese medicine, can provide traditional Chinese medicine ingredients composition and the component analytical approach of activated information one to one but have not yet to see simultaneously.Patent of the present invention is taking miniaturization fixed enzyme electrode as activity rating target, detect the integration of each parts by chromatographic resolution-detecting device-online removal of solvents-enzyme electrode, form the online composition-active appraisal experiment of Chinese medicine device, and utilize this device, carry out the traditional Chinese medicine ingredients-active on-line evaluation application of the bighead atractylodes rhizome.Analysis result will provide traditional Chinese medicine ingredients composition information and the activated information with enzyme electrode effect simultaneously, this on-line analysis result, in conjunction with chemical composition qualification result, bighead atractylodes rhizome composition-activated information will be provided comprehensively, can provide new technical method for setting forth bighead atractylodes rhizome drug action mechanism.This technology can be applied to other Chinese medicine simultaneously, can further strengthen the effective constituent research combining with theory of traditional Chinese medical science, finally set up a kind of new medicine quality evaluated system taking composition, activity as overall target, by strengthening Quality Control, improve Chinese crude drug quality, promote Development of Traditional Chinese Medicine.
Summary of the invention
In order to solve above-mentioned technical matters, an object of the present invention is to provide a kind of liquid chromatography-enzyme electrode combined apparatus, the one-tenth that can obtain in testing sample by this device is grouped into information, and further enzyme corresponding to clearly each component, in conjunction with activity, obtains composition-active on-line evaluation result.Another object of the present invention is to provide the method that adopts above-mentioned device to carry out traditional Chinese medicine ingredients-active on-line analysis.
In order to realize first above-mentioned object, the present invention has adopted following technical scheme:
A kind of liquid chromatography-enzyme electrode combined apparatus, this device comprises chromatographic resolution unit, detecting device, organic solvent removal unit, activity rating unit and computing machine, described chromatographic resolution unit comprises successively by the interconnective mobile phase storage tank of pipeline, chromatogram pump, sampling valve and liquid-phase chromatographic column, the detecting device described in liquid-phase chromatographic column connects by pipeline; Described organic solvent removal unit is for removing the organic solvent of sample, the front end connection detector of organic solvent removal unit; Described activity rating unit comprises mixer, damping fluid storage tank and enzyme electrode, and mixer is connected with the rear end of organic solvent removal unit, and damping fluid storage tank is connected to mixer, and mixer is connected to enzyme electrode; Described computing machine is electrically connected respectively described detecting device and enzyme electrode.
As preferably, described chromatographic resolution unit adopts high performance liquid chromatography piece-rate system or ultrahigh pressure liquid phase chromatographic fractionation system.
As preferably, described detecting device is selected single length ultraviolet detecting device, diode permutation detecting device or fluorescence detector.
As preferably, described organic solvent removal unit comprises nitrogen cylinder, nitrogen pump, heating tube and evaporation tube, nitrogen cylinder connects heating tube by nitrogen pump, described evaporation tube comprises interlayer and lower interlayer, and lower interlayer is mobile phase pipeline, and the front end of mobile phase pipeline is connected with detecting device, upper strata is heated nitrogen pipeline, heated nitrogen pipeline is connected with heating tube, between upper interlayer and lower interlayer, is micropore permeation film, and mobile phase pipeline rear end is connected with mixer.
Formed by voided polytetrafluoroethylene film and hollow-fibre membrane as preferred again, described micropore permeation film.
Adopt conventional nitrogen cylinder as preferred again, described nitrogen cylinder, nitrogen pump discharge range of control: 0.1-2 L/h, pressure: 5-50 PSI, heating tube temperature controlling range: 50-100 DEG C.
In order to realize second above-mentioned object, the present invention has adopted following technical scheme:
The active on-line analysis of a kind of liquid chromatography-enzyme electrode coupling traditional Chinese medicine ingredients, the operation that the method adopts above-mentioned liquid chromatography-enzyme electrode combined apparatus to comprise the following steps:
1) preparation of mobile phase
Employing organic solvent is organic phase, taking water or buffer solution as water, after miillpore filter filters, for subsequent use;
2) pre-service of testing sample
Get testing sample, through operating process such as extraction, solvent extraction, concentrated, constant volume, miillpore filter filtrations, be prepared into need testing solution, for subsequent use;
3) preparation of activity rating unit damping fluid
As required, select suitable buffer solution system, preparation buffer solution, for subsequent use;
4) chromatographic separation condition setting
Connecting on complete liquid chromatography-enzyme electrode combined apparatus, opening computing machine, flow rate of mobile phase is being set, selecting gradient elution that gradient elution program need to be set; Get the mobile phase of step 1), open chromatogram pump and drive mobile phase, enter separable programming;
5) sample introduction
Get step 2) need testing solution, through sampling valve inject chromatographic fractionation system; Separation condition in step 4) is carried out component separating; Open detection device power supply, arranges detecting device detected parameters simultaneously; Need testing solution is by chromatography column, and complicated ingredient is realized post and separated; Composition after separation obtains chromatographic signal by detection, and flows out detecting device and become separated component solution;
6) organic solvent is removed
When chromatographic resolution unit is opened, open organic solvent removal unit, the organic solvent in the purifying composition solution of step 5) is removed, purifying composition is retained in becomes purifying composition aqueous solution in water;
7) enzyme electrode is installed
Buy commercially available small-sized enzyme electrode, or select suitable enzyme, prepare voluntarily fixed enzyme electrode, be installed on activity rating unit, for subsequent use;
8) activity rating
The purification of aqueous solutions of step 6) flows into mixer, gets step 2 simultaneously) damping fluid, in injecting mixer, mix, mixed flow is through the enzyme electrode of step 7), composition and enzyme association reaction, the signals such as generation current or current potential change, and COMPUTER DETECTION signal obtains electrochemical signals;
9) data analysis
The chromatographic signal obtaining according to step 5), and the electrochemical signals that obtains of step 8), by data analysis, the one-tenth obtaining in testing sample is grouped into information, and further enzyme corresponding to clearly each component, in conjunction with activity, obtains composition-active on-line evaluation result.
As preferably, described step 1) organic solvent is selected methyl alcohol or acetonitrile.
The present invention, owing to having adopted above-mentioned experimental provision and applying step, has following feature:
1, design innovation
Existing analytical technology, all taking chemical constitution study as object, cannot provide instant biologically active information.Patent of the present invention is selected the zymoprotein molecule relevant to herbal function, prepare enzyme electrode, as the target of activity rating, the biologically active of each composition in analytic sample can be provided online, in real time, the one-tenth in Realization analysis testing sample is grouped into and biologically active corresponding to the each component of on-line evaluation.Enzyme electrode is combined liquid phase analysis, can break through the disappearance of composition activated information in existing analytical technology, provides thinking and technical scheme for setting up new medicine quality evaluated pattern.
2, application prospect and function well
This technology can be applied to other Chinese medicine, can further strengthen the effective constituent research combining with theory of traditional Chinese medical science, finally set up a kind of new medicine quality evaluated system taking composition, activity as overall target, by strengthening Quality Control, improve Chinese crude drug quality, promote Development of Traditional Chinese Medicine.
Brief description of the drawings
Fig. 1 device schematic diagram of the present invention.
Fig. 2 composition-active on-line evaluation pattern diagram.
Fig. 3 bighead atractylodes rhizome alcohol extract HPLC figure.
Fig. 4 bighead atractylodes rhizome composition enzyme electrode detection signal figure, a, b, c, d is followed successively by atractylenolide, III, I and atractylone.
Embodiment
embodiment 1
A kind of liquid chromatography-enzyme electrode combined apparatus as shown in Figure 1, this device comprises chromatographic resolution unit, detecting device, organic solvent removal unit, activity rating unit and computing machine 12, chromatographic resolution unit adopts high performance liquid chromatography piece-rate system or ultrahigh pressure liquid phase chromatographic fractionation system, and detecting device is selected single length ultraviolet detecting device, diode permutation detecting device or fluorescence detector.Described chromatographic resolution unit comprises successively by the interconnective mobile phase storage tank 1 of pipeline, chromatogram pump 2, sampling valve 3 and liquid-phase chromatographic column 4, the detecting device 5 described in liquid-phase chromatographic column 4 connects by pipeline.Described organic solvent removal unit comprises nitrogen cylinder, nitrogen pump 7, heating tube 8 and evaporation tube 6, nitrogen cylinder adopts conventional nitrogen cylinder, nitrogen pump 7 flow control scopes: 0.1-2 L/h, pressure: 5-50 PSI, heating tube 8 temperature controlling range: 50-100 DEG C.Nitrogen cylinder connects heating tube 8 by nitrogen pump 7, described evaporation tube 6 comprises interlayer and lower interlayer, lower interlayer is mobile phase pipeline, the front end of mobile phase pipeline is connected with detecting device 5, upper strata is heated nitrogen pipeline, heated nitrogen pipeline is connected with heating tube 8, between upper interlayer and lower interlayer, is micropore permeation film, and micropore permeation film is made up of voided polytetrafluoroethylene film and hollow-fibre membrane.Described activity rating unit comprises mixer 9, damping fluid storage tank 10 and enzyme electrode 11, and the rear end that mixer 9 connects mobile phase pipeline is connected, and damping fluid storage tank 10 is connected to mixer 9, and mixer 9 is connected to enzyme electrode 11; Described computing machine 12 is electrically connected respectively described detecting device 5 and enzyme electrode 11.
embodiment 2
The active on-line analysis of a kind of liquid chromatography-enzyme electrode coupling traditional Chinese medicine ingredients, the operation that the method adopts liquid chromatography-enzyme electrode combined apparatus of embodiment 1 to comprise the following steps:
1) preparation of mobile phase
Taking water as water, methyl alcohol is organic phase.Remove ion pure water and chromatogram methyl alcohol, cross respectively the miillpore filter in 0.45 μ aperture, filtrate is placed in respectively water and organic phase solvent bottle, for subsequent use.
2) pre-service of testing sample
Get dry Rhizoma Atractylodis Macrocephalae, pulverize, take 0.6 g, be placed in 25 mL volumetric flasks, add 20 mL methyl alcohol, ultrasonic extraction 20min, take out, put to room temperature, add chromatogram methyl alcohol to scale, shake up, extract filters through 0.22 μ miillpore filter, subsequent filtrate is placed in sample injection bottle, is prepared into need testing solution, for subsequent use.
3) preparation of activity rating unit damping fluid
Select the approaching damping fluid of PH and simulated intestinal fluid, take potassium dihydrogen phosphate 6.8 g, be placed in 1000 mL volumetric flasks, 500 mL that add water make to dissolve, and with 0.1 mol/L sodium hydroxide solution adjusting pH value to 6.8, add water to scale, shake up, and obtain damping fluid, for subsequent use.
4) chromatographic separation condition setting
Liquid chromatographic system is selected LC-20A-VP liquid chromatograph (Japanese Shimadzu), and CAPCELL PAK C18 analysis chromatographic column (250 mm × 4.6 mm, 5 μ m), connect the each parts of experimental provision; Open computer power supply, it is 1.0 mL/min that flow rate of mobile phase is set, and gradient elution program is in Table; Get the mobile phase of step 1), chromatogram pump water and organic phase pump head are placed in to solvent bottle separately, open chromatogram pump power supply, drive mobile phase with the initial ratio of gradient elution program, after system balancing 20 min, move gradient elution program.
Gradient elution program
5) sample introduction
Sample introduction when gradient elution program brings into operation, the accurate step 2 of drawing) need testing solution 10 μ L, inject chromatographic fractionation system through sampling valve; Separation condition in step 4) is carried out component separating; Simultaneously open detection device 5 power supplys, detecting device 5 is set, and to detect wavelength be 220 nm; Need testing solution is by chromatography column, and complicated ingredient is realized post and separated; Composition after separation is detected and is obtained chromatographic signal (seeing Figure of description 3) by detecting device 5, and each component flows out detecting device 5 successively becomes separated component solution;
6) organic solvent is removed
When chromatographic resolution unit is opened, open nitrogen pump 7, it is 0.6 L/h that nitrogen flow rate is set, reduction valve pressure 30 PSI; Open heating tube power supply, it is 85 DEG C that heating-up temperature is set; The separated component of step 5) is 4 kinds of one-components, the evaporation tube 6 of flowing through successively, and organic solvent evaporates into gaseous state and sees through and remove through permeable membrane, and separated component is retained in becomes purifying composition aqueous solution in water;
7) enzyme electrode 11 is installed
Get the small-sized enzyme electrode 11 of commercially available acetylcholine Esterified Enzyme, be installed on activity rating unit; Enzyme electrode 11 is active detector 5, and selection potential range is 0.45 ~ 0.75V, the Ampere currents signal of research purifying composition on electrode and the graph of a relation of operating potential.When current potential is during higher than 0.65 V, peak current increasess slowly and substrate current still has increase, and best effort current potential is chosen in 0.65 V, to ensure higher signal to noise ratio (S/N ratio);
8) activity rating
The purifying composition aqueous solution of step 6) flows into mixer 9, get step 2 simultaneously) damping fluid, in injecting mixer 9, mix, mixed flow is through the enzyme electrode 11 of step 7), composition and enzyme association reaction, the variation generation current relevant with operating potential changes, and COMPUTER DETECTION signal obtains the graph of a relation of Ampere currents signal and operating potential, sees Figure of description 4;
9) data analysis
The chromatographic signal obtaining according to step 5), in extract, 4 kinds of single components are followed successively by: atractylenolide, III, I and atractylone; The electrochemical signals that step 8) obtains, shows that atractylone is combined activity with acetylcholine Esterified Enzyme the strongest, and atractylenolide, I and III activity reduce successively.
Claims (8)
1. liquid chromatography-enzyme electrode combined apparatus, this device comprises chromatographic resolution unit, detecting device, organic solvent removal unit, activity rating unit and computing machine (12), it is characterized in that: chromatographic resolution unit comprises successively by the interconnective mobile phase storage tank of pipeline (1), chromatogram pump (2), sampling valve (3) and liquid-phase chromatographic column (4), the detecting device (5) described in liquid-phase chromatographic column (4) connects by pipeline; Described organic solvent removal unit is for removing the organic solvent of sample, the front end connection detector (5) of organic solvent removal unit; Described activity rating unit comprises mixer (9), damping fluid storage tank (10) and enzyme electrode (11), mixer (9) connects and is connected with the rear end of organic solvent removal unit, damping fluid storage tank (10) is connected to mixer (9), and mixer (9) is connected to enzyme electrode (11); Described computing machine (12) is electrically connected respectively described detecting device (5) and enzyme electrode (11).
2. a kind of liquid chromatography-enzyme electrode combined apparatus according to claim 1, is characterized in that: described chromatographic resolution unit adopts high performance liquid chromatography piece-rate system or ultrahigh pressure liquid phase chromatographic fractionation system.
3. a kind of liquid chromatography-enzyme electrode combined apparatus according to claim 1, is characterized in that: detecting device (5) is selected single length ultraviolet detecting device (5), diode permutation detecting device (5) or fluorescence detector (5).
4. according to a kind of liquid chromatography-enzyme electrode combined apparatus described in claim 1 or 2 or 3, it is characterized in that: described organic solvent removal unit comprises nitrogen cylinder, nitrogen pump (7), heating tube (8) and evaporation tube (6), nitrogen cylinder connects heating tube (8) by nitrogen pump, described evaporation tube (6) comprises interlayer and lower interlayer, lower interlayer is mobile phase pipeline, the front end of mobile phase pipeline is connected with detecting device (5), upper strata is heated nitrogen pipeline, heated nitrogen pipeline is connected with heating tube (8), between upper interlayer and lower interlayer, it is micropore permeation film, mobile phase pipeline rear end is connected with mixer (9).
5. a kind of liquid chromatography-enzyme electrode combined apparatus according to claim 4, is characterized in that: micropore permeation film is made up of voided polytetrafluoroethylene film and hollow-fibre membrane.
6. a kind of liquid chromatography-enzyme electrode combined apparatus according to claim 4, it is characterized in that: nitrogen cylinder adopts conventional nitrogen cylinder, nitrogen pump discharge range of control: 0.1-2L/h, pressure: 5-50PSI, heating tube (8) temperature controlling range: 50-100 DEG C.
7. the active on-line analysis of liquid chromatography-enzyme electrode coupling traditional Chinese medicine ingredients, is characterized in that: the operation that the method adopts the liquid chromatography-enzyme electrode combined apparatus described in claim 1 ~ 6 any one claim to comprise the following steps:
1) preparation of mobile phase
Employing organic solvent is organic phase, taking water or buffer solution as water, after miillpore filter filters, for subsequent use;
2) pre-service of testing sample
Get testing sample, through operating process such as extraction, solvent extraction, concentrated, constant volume, miillpore filter filtrations, be prepared into need testing solution, for subsequent use;
3) preparation of activity rating unit damping fluid
As required, select suitable buffer solution system, preparation buffer solution, for subsequent use;
4) chromatographic separation condition setting
Connecting on complete liquid chromatography-enzyme electrode combined apparatus, opening computing machine, flow rate of mobile phase is being set, selecting gradient elution that gradient elution program need to be set; Get the mobile phase of step 1), open chromatogram pump and drive mobile phase, enter separable programming;
5) sample introduction
Get step 2) need testing solution, through sampling valve inject chromatographic fractionation system; Separation condition in step 4) is carried out component separating; Open detection device (5) power supply, arranges detecting device (5) detected parameters simultaneously; Need testing solution is by chromatography column, and complicated ingredient is realized post and separated; Composition after separation obtains chromatographic signal by detection, and flows out detecting device (5) and become separated component solution;
6) organic solvent is removed
When chromatographic resolution unit is opened, open organic solvent removal unit, the organic solvent in the purifying composition solution of step 5) is removed, purifying composition is retained in becomes purifying composition aqueous solution in water;
7) enzyme electrode (11) is installed
Buy commercially available small-sized enzyme electrode (11), or select suitable enzyme, prepare voluntarily fixed enzyme electrode (11), be installed on activity rating unit, for subsequent use;
8) activity rating
The purification of aqueous solutions of step 6) flows into mixer (9), get step 2 simultaneously) damping fluid, in injecting mixer (9), mix, mixed flow is through the enzyme electrode (11) of step 7), composition and enzyme association reaction, the signal such as generation current or current potential changes, and COMPUTER DETECTION signal obtains electrochemical signals;
9) data analysis
The chromatographic signal obtaining according to step 5), and the electrochemical signals that obtains of step 8), by data analysis, the one-tenth obtaining in testing sample is grouped into information, and further enzyme corresponding to clearly each component, in conjunction with activity, obtains composition-active on-line evaluation result.
8. the active on-line analysis of a kind of liquid chromatography-enzyme electrode coupling traditional Chinese medicine ingredients according to claim 7, is characterized in that: step 1) organic solvent is selected methyl alcohol or acetonitrile.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410248217.4A CN104007219B (en) | 2014-06-05 | 2014-06-05 | A kind of liquid chromatography-enzyme electrode combined apparatus and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410248217.4A CN104007219B (en) | 2014-06-05 | 2014-06-05 | A kind of liquid chromatography-enzyme electrode combined apparatus and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104007219A true CN104007219A (en) | 2014-08-27 |
| CN104007219B CN104007219B (en) | 2016-03-30 |
Family
ID=51367973
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410248217.4A Active CN104007219B (en) | 2014-06-05 | 2014-06-05 | A kind of liquid chromatography-enzyme electrode combined apparatus and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104007219B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109313169A (en) * | 2016-05-06 | 2019-02-05 | 德克萨斯系统大学董事会 | The preparation of volatility eluent |
| CN109580857A (en) * | 2018-12-29 | 2019-04-05 | 北京中研同仁堂医药研发有限公司 | Rhizoma Atractylodis Macrocephalae characteristic spectrum construction method and quality determining method |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0104935A2 (en) * | 1982-09-28 | 1984-04-04 | The Yellow Springs Instrument Company, Inc. | Liquid chromatograph enzyme electrode detector |
| JPH0593714A (en) * | 1991-09-30 | 1993-04-16 | Shimadzu Corp | Lc-ms interface |
| WO1994020841A1 (en) * | 1993-03-05 | 1994-09-15 | University Of Wollongong | Pulsed electrochemical detection using polymer electroactive electrodes |
| JP2003172727A (en) * | 2001-09-28 | 2003-06-20 | Jasco Corp | High speed liquid chromatograph device |
| CN1764493A (en) * | 2003-01-30 | 2006-04-26 | 迪奥尼克斯公司 | Capacity chemical suppressors and method of use |
| CN101542292A (en) * | 2006-09-22 | 2009-09-23 | 爱科来株式会社 | Blood analysis apparatus |
-
2014
- 2014-06-05 CN CN201410248217.4A patent/CN104007219B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0104935A2 (en) * | 1982-09-28 | 1984-04-04 | The Yellow Springs Instrument Company, Inc. | Liquid chromatograph enzyme electrode detector |
| JPH0593714A (en) * | 1991-09-30 | 1993-04-16 | Shimadzu Corp | Lc-ms interface |
| WO1994020841A1 (en) * | 1993-03-05 | 1994-09-15 | University Of Wollongong | Pulsed electrochemical detection using polymer electroactive electrodes |
| JP2003172727A (en) * | 2001-09-28 | 2003-06-20 | Jasco Corp | High speed liquid chromatograph device |
| CN1764493A (en) * | 2003-01-30 | 2006-04-26 | 迪奥尼克斯公司 | Capacity chemical suppressors and method of use |
| CN101542292A (en) * | 2006-09-22 | 2009-09-23 | 爱科来株式会社 | Blood analysis apparatus |
Non-Patent Citations (3)
| Title |
|---|
| F.ORTEGA ET AL.: "Liquid chromatographic separation of phenolic drugs using catalytic detection: comparison of an enzyme reactor and enzyme electrode", 《JOURNAL OF PHARMACEUTICAL & BIOMEDICAL ANALYSIS》, vol. 10, 31 December 1992 (1992-12-31) * |
| SUSUMU YAMATO ET AL.: "Specific detection of acetyl-coenzyme A by reversed-phase ion-pair high-performance liquid chromatography with an immobilized enzyme reactor", 《JOURNAL OF CHROMATOGRAPHY》, vol. 590, 31 December 1992 (1992-12-31), pages 241 - 245, XP026538908, DOI: 10.1016/0021-9673(92)85387-9 * |
| 干宁 等: "采用纳米修饰双酶电极生物传感器检测有机膦与氨基甲酸酯类农药", 《农药学学报》, vol. 10, no. 3, 31 December 2008 (2008-12-31) * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109313169A (en) * | 2016-05-06 | 2019-02-05 | 德克萨斯系统大学董事会 | The preparation of volatility eluent |
| CN109313169B (en) * | 2016-05-06 | 2020-12-01 | 德克萨斯系统大学董事会 | Volatile eluent preparation |
| CN109580857A (en) * | 2018-12-29 | 2019-04-05 | 北京中研同仁堂医药研发有限公司 | Rhizoma Atractylodis Macrocephalae characteristic spectrum construction method and quality determining method |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104007219B (en) | 2016-03-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP7177103B2 (en) | System and method for two-dimensional RPLC-SFC chromatography | |
| Fu et al. | Screening techniques for the identification of bioactive compounds in natural products | |
| Venkatramani et al. | Simultaneous achiral-chiral analysis of pharmaceutical compounds using two-dimensional reversed phase liquid chromatography-supercritical fluid chromatography | |
| Kataoka | New trends in sample preparation for clinical and pharmaceutical analysis | |
| Ali et al. | Hyphenation in sample preparation: advancement from the micro to the nano world | |
| Chen et al. | A single-column procedure on Bond Elut Certify for systematic toxicological analysis of drugs in plasma and urine | |
| CA2685358C (en) | Method of detecting blood plasma danshensu and salvianolic acid b after administration of fuzheng huayu (fzhy) | |
| CN102621244A (en) | Construction method for HPLC (high performance liquid chromatography) finger-print chromatogram of ginseng and astragalus strengthening injection and application of finger-print | |
| Tang et al. | Sample preparation for analyzing traditional Chinese medicines | |
| CN106267897A (en) | Ginsenoside and the method for residual pesticide in sharp separation Radix Ginseng | |
| CN110208401A (en) | Solid phase is dehydrated extraction-supercritical fluid chromatography-mass spectrum on-line analysis system and method | |
| Fu et al. | Chemical separation and characterization of complex samples with herbal medicine | |
| US20100173345A1 (en) | Detection of blood plasma amygdalin of dissipating blood stasis botanical | |
| Yan et al. | Electromembrane extraction of aristolochic acids: New insights in separation of bioactive ingredients of traditional Chinese medicines | |
| Shen et al. | Hyphenation of optimized microfluidic sample preparation with nano liquid chromatography for faster and greener alkaloid analysis | |
| CN104007219B (en) | A kind of liquid chromatography-enzyme electrode combined apparatus and application thereof | |
| Zhang et al. | On‐line hyphenation of supercritical fluid extraction and two‐dimensional high performance liquid chromatography‐atmospheric pressure chemical ionization tandem mass spectrometer for the analysis of Ganoderma lucidum | |
| Zhao et al. | Quantitative determination of amphetamine-type stimulants in sewage and urine by hybrid monolithic column solid-phase microextraction coupled with UPLC-QTRAP MS/MS | |
| Liu et al. | Quantitative Determination of 7 Saikosaponins in Xiaochaihu Granules Using High‐Performance Liquid Chromatography with Charged Aerosol Detection | |
| Yao et al. | Ten years of research into phytomedicines analysis-an era in new technologies and methods | |
| Zhang et al. | Coupling of micro solid-phase extraction with electrospray ionization and its potential for complex sample analyses using a miniature mass spectrometer | |
| Majors | Application of existing sample preparation technologies to new areas | |
| Shen | New analytical approaches for faster or greener phytochemical analyses | |
| CN113834879A (en) | Establishment and application of epimedium herb flavone component fingerprint spectrum based on SPE technology | |
| acids Acetone | CUMULATIVE COMPOUND INDEX J. CHROMATOGR. A VOLS. 952–1000 |
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 |