CN104007219B - A kind of liquid chromatography-enzyme electrode combined apparatus and application thereof - Google Patents
A kind of liquid chromatography-enzyme electrode combined apparatus and application thereof Download PDFInfo
- Publication number
- CN104007219B CN104007219B CN201410248217.4A CN201410248217A CN104007219B CN 104007219 B CN104007219 B CN 104007219B CN 201410248217 A CN201410248217 A CN 201410248217A CN 104007219 B CN104007219 B CN 104007219B
- Authority
- CN
- China
- Prior art keywords
- enzyme electrode
- organic solvent
- liquid chromatography
- mixer
- 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.)
- Active
Links
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention relates to 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, 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 is connected by pipeline; Organic solvent removal unit for removing the organic solvent in sample, the front end connection detector of organic solvent removal unit; 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; Computing machine is electrically connected described detecting device and enzyme electrode respectively.The present invention can provide the biologically active analyzing each composition in sample online, in real time, and the one-tenth in Realization analysis testing sample is grouped into the biologically active that also each component of on-line evaluation is corresponding.
Description
Technical field
The present invention relates to one and belong to drug world, particularly relate 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 achieves very big progress, and the object of Analysis of Chinese Traditional Medicine more concentrates on for the key scientific problems in TCM Modernization research; Analysis of Chinese Traditional Medicine needs various new method, new technology integration to make it to become the new system solving key scientific problems simultaneously, and is no longer confined to the material composition analyzing Chinese medicine, and what more focus on solution is the composition of material and the correlativity of function thereof.The development of total system biology (GlobalSystemsBiology), system disclose that the change of chemical matterome and biosystem reply time null response correlativity; And imagine for the research of the Chinese medicine compound prescription metabolism of marker-drug effect target three dimensional pattern based on the square medicine fingerprint-medicine of metabolism group, all embody and improve 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 chromatogram-nuclear-magnetism coupling are developed, abundant structural information is provided while centering prodrug complex composition carries out compartment analysis, achieve qualitative and quantitative analysis that is known in Chinese medicine or non-principal component, greatly advance traditional Chinese medicine ingredients chemical research, but existing analytical technology is all for the purpose of chemical constitution study, cannot react online or the biologically active of evaluative component.Therefore, select the zymoprotein molecule relevant to herbal function, fixed enzyme membrane is made by physical chemistry crosslinked action immobilization, then enzyme electrode (enzymeelectrode) is bonded with electrochemical sensor, utilize the selectivity of enzyme reaction, stability and electrochemical analysis quick, sensitive, the biologically active analyzing each composition in sample is provided online, in real time.Enzyme electrode associating liquid phase analysis, can breaking through the disappearance of Ingredients Active information in existing analytical technology, providing thinking and technical scheme for setting up new medicine quality evaluated pattern.
Electrode analysis method (comprising enzyme electrode), as a kind of electrochemical means of detection, successfully realizes 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, then can realize the coupling of liquid chromatography and enzyme electrode.
In stratographic analysis, enrichment method is the conventional pre-treatment means of sample, namely enters before chromatographic column is separated to carry out enrichment method at sample.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, permission organic solvent permeates, removal of solvents can be realized, and reach the concentrated object of sample component, for further activity rating provides basis.Organic solvent removal device designs: when mobile phase is by porous permeable film pipe, flows through in tube wall periphery the dry gas (such as air or nitrogen) heated, and organic solvent is heated volatilization for gaseous state, removes through permeable membrane.
Traditional Chinese medicine ingredients composition is complicated, and conventional analytical technology has played Chinese medicine effect in the qualitative and quantitative analysis of Chinese medicine, but has not yet to see traditional Chinese medicine ingredients can be provided simultaneously to form the analytical approach with component activated information one to one.Patent of the present invention with miniaturization fixed enzyme electrode for activity rating target, the integration of each parts is detected by chromatographic resolution-detecting device-online removal of solvents-enzyme electrode, form Chinese medicine online composition-active appraisal experiment device, and utilize this device, carry out the traditional Chinese medicine ingredients-active on-line evaluation application of the bighead atractylodes rhizome.Analysis result forms information and the activated information with enzyme electrode effect by providing traditional Chinese medicine ingredients simultaneously, this on-line analysis result, in conjunction with chemical composition qualification result, bighead atractylodes rhizome composition-activated information being provided comprehensively, new technical method can be provided for setting forth bighead atractylodes rhizome drug action mechanism.This technology can be applied to other Chinese medicine simultaneously, the effective constituent research combined with theory of traditional Chinese medical science can be strengthened further, finally set up a kind of medicine quality evaluated system being overall target with composition, activity newly, 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 be obtained in testing sample by this device is grouped into information, and the enzyme binding activities that clearly each component is corresponding further, obtain composition-active on-line evaluation result.Another object of the present invention is to provide the method adopting above-mentioned device to carry out traditional Chinese medicine ingredients-active on-line analysis.
In order to realize first above-mentioned object, present invention employs 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 is connected by pipeline; Described organic solvent removal unit for removing the organic solvent in 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 described detecting device and enzyme electrode respectively.
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 selects Single wavelength UV-detector, 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, and be micropore permeation film between upper interlayer and lower interlayer, mobile phase pipeline rear end is connected with mixer.
As preferred again, described micropore permeation film is made up of voided polytetrafluoroethylene film and hollow-fibre membrane.
As preferred again, described nitrogen cylinder adopts conventional nitrogen cylinder, nitrogen pump discharge range of control: 0.1-2L/h, pressure: 5-50PSI, heating tube temperature range of control: 50-100 DEG C.
In order to realize second above-mentioned object, present invention employs following technical scheme:
The active on-line analysis of a kind of liquid chromatography-enzyme electrode coupling traditional Chinese medicine ingredients, the method adopts above-mentioned liquid chromatography-enzyme electrode combined apparatus to carry out the operation comprised the following steps:
1) preparation of mobile phase
Employing organic solvent is organic phase, with water or buffer solution for aqueous phase, 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 is arranged
Connecting on complete liquid chromatography-enzyme electrode combined apparatus, opening computing machine, flow rate of mobile phase is set, selecting gradient elution to need to arrange gradient elution program; 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; Component separating is carried out in the separation condition of step 4); Open detection device power supply, arranges detecting device detected parameters simultaneously; Need testing solution is by chromatography column, and complicated ingredient realizes post and is separated; Composition after separation obtains chromatographic signal by detection, and flows out detecting device and become separated component solution;
6) organic solvent is removed
Chromatographic resolution unit opens organic solvent removal unit while opening, and removed by the organic solvent in the purified fractions solution of step 5), purified fractions is retained in aqueous phase becomes purified fractions aqueous solution;
7) enzyme electrode is installed
Buy commercially available small-sized enzyme electrode, or select suitable enzyme, prepare fixed enzyme electrode voluntarily, 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, mix in injecting mixer, mixed flow is through the enzyme electrode 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
According to the chromatographic signal that step 5) obtains, and the electrochemical signals that step 8) obtains, by data analysis, the one-tenth obtained in testing sample is grouped into information, and the enzyme binding activities that clearly each component is corresponding further, obtain composition-active on-line evaluation result.
As preferably, described step 1) organic solvent selects methyl alcohol or acetonitrile.
The present invention, owing to have employed above-mentioned experimental provision and applying step, has following feature:
1, design innovation
Existing analytical technology, all for the purpose of chemical constitution study, cannot provide instant biologically active information.Patent of the present invention selects the zymoprotein molecule relevant to herbal function, prepare enzyme electrode, as the target of activity rating, can provide the biologically active analyzing each composition in sample online, in real time, the one-tenth in Realization analysis testing sample is grouped into the biologically active that also each component of on-line evaluation is corresponding.Enzyme electrode associating liquid phase analysis, can breaking through the disappearance of Ingredients Active information in existing analytical technology, providing 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, the effective constituent research combined with theory of traditional Chinese medical science can being strengthened further, finally setting up a kind of medicine quality evaluated system being overall target with composition, activity newly, by strengthening Quality Control, improve Chinese crude drug quality, promote Development of Traditional Chinese Medicine.
Accompanying drawing explanation
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 schemes.
Fig. 4 bighead atractylodes rhizome composition enzyme electrode detection signal figure, a, b, c, d be 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 selects Single wavelength UV-detector, 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 is connected 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 scope: 0.1-2L/h, pressure: 5-50PSI, 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, and be micropore permeation film between upper interlayer and lower interlayer, 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 described detecting device 5 and enzyme electrode 11 respectively.
embodiment 2
The active on-line analysis of a kind of liquid chromatography-enzyme electrode coupling traditional Chinese medicine ingredients, the method adopts the liquid chromatography-enzyme electrode combined apparatus of embodiment 1 to carry out the operation comprised the following steps:
1) preparation of mobile phase
Take water as aqueous phase, methyl alcohol is organic phase.Remove ion pure water and chromatogram methyl alcohol, cross the miillpore filter in 0.45 μ aperture respectively, filtrate is placed in aqueous phase and organic phase solvent bottle respectively, for subsequent use.
2) pre-service of testing sample
Get dry Rhizoma Atractylodis Macrocephalae, pulverize, take 0.6g, be placed in 25mL volumetric flask, add 20mL methyl alcohol, ultrasonic extraction 20min, takes out, puts to room temperature, add chromatogram methyl alcohol to scale, shake up, extract filters through 0.22 μ miillpore filter, and 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 damping fluid that PH and simulated intestinal fluid are close, take potassium dihydrogen phosphate 6.8g, be placed in 1000mL volumetric flask, the 500mL that adds water makes dissolving, with 0.1mol/L sodium hydroxide solution adjustment pH value to 6.8, adds water to scale, shakes up, obtain damping fluid, for subsequent use.
4) chromatographic separation condition is arranged
Liquid chromatographic system selects LC-20A-VP liquid chromatograph (Japanese Shimadzu), and CAPCELLPAKC18 analyzes chromatographic column (250mm × 4.6mm, 5 μm), connects each parts of experimental provision; Open computer power supply, arranging flow rate of mobile phase is 1.0mL/min, and gradient elution program is in Table; Get the mobile phase of step 1), chromatogram pump aqueous phase and organic phase pump head are placed in respective solvent bottle, open chromatogram pump power supply, drive mobile phase with gradient elution program original ratio, after system balancing 20min, run gradient elution program.
Gradient elution program
5) sample introduction
Sample introduction when gradient elution program brings into operation, accurate aspiration step 2) need testing solution 10 μ L, inject chromatographic fractionation system through sampling valve; Component separating is carried out in the separation condition of step 4); Open detection device 5 power supply simultaneously, arranging detecting device 5 determined wavelength is 220nm; Need testing solution is by chromatography column, and complicated ingredient realizes post and is separated; Composition after separation is detected by detecting device 5 and obtains chromatographic signal (see Figure of description 3), and each component flows out detecting device 5 successively becomes separated component solution;
6) organic solvent is removed
Chromatographic resolution unit opens nitrogen pump 7 while opening, and arranging nitrogen flow rate is 0.6L/h, reduction valve pressure 30PSI; Open heating tube power supply, arranging heating-up temperature is 85 DEG C; The separated component of step 5) is 4 kinds of one-components, and flow through evaporation tube 6 successively, organic solvent evaporates into gaseous state through permeable membrane through removal, and separated component is retained in aqueous phase becomes purified fractions aqueous solution;
7) enzyme electrode 11 is installed
Get the small-sized enzyme electrode 11 of commercially available acetylcholine esterification enzyme, be installed on activity rating unit; Enzyme electrode 11 is active detector 5, and selection potential range is 0.45 ~ 0.75V, the research amp current signal of purified fractions on electrode and the graph of a relation of operating potential.When current potential is higher than 0.65V, peak current increasess slowly and substrate current still has increase, and best effort current potential is selected at 0.65V, to ensure higher signal to noise ratio (S/N ratio);
8) activity rating
The purified fractions aqueous solution of step 6) flows into mixer 9, get step 2 simultaneously) damping fluid, mix in injecting mixer 9, mixed flow is through the enzyme electrode 11 of step 7), composition and enzyme association reaction, with the change generation current change that operating potential is relevant, COMPUTER DETECTION signal obtains the graph of a relation of amp current signal and operating potential, sees Figure of description 4;
9) data analysis
According to the chromatographic signal that step 5) obtains, in extract, 4 kinds of single components are followed successively by: atractylenolide, III, I and atractylone; The electrochemical signals that step 8) obtains, the most by force, atractylenolide, I and III activity reduce successively for display atractylone and acetylcholine esterification enzyme binding activities.
Claims (7)
1. the active on-line analysis of liquid chromatography-enzyme electrode coupling traditional Chinese medicine ingredients, the method adopts 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 (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) is connected by pipeline; Described organic solvent removal unit for removing the organic solvent in 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) 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 described detecting device (5) and enzyme electrode (11) respectively;
The operation that the method comprises the following steps:
1) preparation of mobile phase adopts organic solvent to be organic phase, with water or buffer solution for aqueous phase, after miillpore filter filters, for subsequent use;
2) testing sample is got in the pre-service of testing sample, through extraction, solvent extraction, concentrated, constant volume, miillpore filter filtration operating process, is prepared into need testing solution, for subsequent use;
3) preparation of activity rating unit damping fluid as required, selects suitable buffer solution system, preparation buffer solution, for subsequent use;
4) chromatographic separation condition is arranged on and connects on complete liquid chromatography-enzyme electrode combined apparatus, opens computing machine, arranges flow rate of mobile phase, selects gradient elution to need to arrange gradient elution program; Get the mobile phase of step 1), open chromatogram pump and drive mobile phase, enter separable programming;
5) sample introduction gets step 2) need testing solution, through sampling valve inject chromatographic fractionation system; Component separating is carried out in the separation condition of step 4); Open detection device (5) power supply, arranges detecting device (5) detected parameters simultaneously; Need testing solution is by chromatography column, and complicated ingredient realizes post and is 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 while chromatographic resolution unit is opened and is opened organic solvent removal unit, and removed by the organic solvent in the separated component solution of step 5), purified fractions is retained in aqueous phase becomes purified fractions aqueous solution;
7) enzyme electrode (11) is installed and is bought commercially available small-sized enzyme electrode (11), or selects suitable enzyme, prepares fixed enzyme electrode (11) voluntarily, is installed on activity rating unit, for subsequent use;
8) the purified fractions aqueous solution of activity rating step 6) flows into mixer (9), get the damping fluid of step 3) simultaneously, mixing in injecting mixer (9), mixed flow is through the enzyme electrode (11) of step 7), composition and enzyme association reaction, generation current or electric potential signal change, and COMPUTER DETECTION signal obtains electrochemical signals;
9) chromatographic signal that obtains according to step 5) of data analysis, and the electrochemical signals that step 8) obtains, by data analysis, the one-tenth obtained in testing sample is grouped into information, and the enzyme binding activities that clearly each component is corresponding further, obtain composition-active on-line evaluation result.
2. the active on-line analysis of a kind of liquid chromatography according to claim 1-enzyme electrode coupling traditional Chinese medicine ingredients, 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. the active on-line analysis of a kind of liquid chromatography according to claim 1-enzyme electrode coupling traditional Chinese medicine ingredients, is characterized in that: detecting device (5) selects Single wavelength UV-detector (5), diode array detector (5) or fluorescence detector (5).
4. the active on-line analysis of a kind of liquid chromatography according to claim 1 or 2 or 3-enzyme electrode coupling traditional Chinese medicine ingredients, 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), be micropore permeation film between upper interlayer and lower interlayer, mobile phase pipeline rear end is connected with mixer (9).
5. the active on-line analysis of a kind of liquid chromatography according to claim 4-enzyme electrode coupling traditional Chinese medicine ingredients, is characterized in that: micropore permeation film is made up of voided polytetrafluoroethylene film and hollow-fibre membrane.
6. the active on-line analysis of a kind of liquid chromatography according to claim 4-enzyme electrode coupling traditional Chinese medicine ingredients, 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 a kind of liquid chromatography according to claim 1-enzyme electrode coupling traditional Chinese medicine ingredients, is characterized in that: step 1) organic solvent selects 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 CN104007219A (en) | 2014-08-27 |
| CN104007219B true 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) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US10228355B2 (en) * | 2016-05-06 | 2019-03-12 | Board Of Regents, The University Of Texas System | 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 |
Citations (4)
| 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 |
| WO1994020841A1 (en) * | 1993-03-05 | 1994-09-15 | University Of Wollongong | Pulsed electrochemical detection using polymer electroactive electrodes |
| 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 |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0593714A (en) * | 1991-09-30 | 1993-04-16 | Shimadzu Corp | Lc-ms interface |
| JP2003172727A (en) * | 2001-09-28 | 2003-06-20 | Jasco Corp | High-performance liquid chromatograph |
-
2014
- 2014-06-05 CN CN201410248217.4A patent/CN104007219B/en active Active
Patent Citations (4)
| 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 |
| WO1994020841A1 (en) * | 1993-03-05 | 1994-09-15 | University Of Wollongong | Pulsed electrochemical detection using polymer electroactive electrodes |
| 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 |
|---|
| Liquid chromatographic separation of phenolic drugs using catalytic detection: comparison of an enzyme reactor and enzyme electrode;F.ORTEGA et al.;《Journal of Pharmaceutical & Biomedical Analysis》;19921231;第10卷;第790-791页"Equipment"节，Figure 1 * |
| Specific detection of acetyl-coenzyme A by reversed-phase ion-pair high-performance liquid chromatography with an immobilized enzyme reactor;Susumu Yamato et al.;《Journal of Chromatography》;19921231;第590卷;Page 241-245 * |
| 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;干宁 等;《农药学学报》;20081231;第10卷(第3期);第332页2.3节 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104007219A (en) | 2014-08-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Venkatramani et al. | Simultaneous achiral-chiral analysis of pharmaceutical compounds using two-dimensional reversed phase liquid chromatography-supercritical fluid chromatography | |
| JP2020091294A (en) | System and method for two-dimensional RPLC-SFC chromatography | |
| Li et al. | Two‐dimensional liquid chromatography and its application in traditional Chinese medicine analysis and metabonomic investigation | |
| CA2685358C (en) | Method of detecting blood plasma danshensu and salvianolic acid b after administration of fuzheng huayu (fzhy) | |
| Chen et al. | A single-column procedure on Bond Elut Certify for systematic toxicological analysis of drugs in plasma and urine | |
| Xie et al. | Extraction and isolation of flavonoid glycosides from Flos Sophorae Immaturus using ultrasonic‐assisted extraction followed by high‐speed countercurrent chromatography | |
| Luo et al. | Molecularly imprinted polymer solid‐phase microextraction coupled with ultra high performance liquid chromatography and tandem mass spectrometry for rapid analysis of pyrrolizidine alkaloids in herbal medicine | |
| Yin et al. | Determination of hyperoside and isoquercitrin in rat plasma by membrane‐protected micro‐solid‐phase extraction with high‐performance liquid chromatography | |
| CN109856278B (en) | Method for screening active ingredients of traditional Chinese medicine based on three-phase laminar flow microfluidic chip | |
| CA2685359C (en) | Method of detecting blood plasma amygdalin after administration of fuzheng huayu(fzhy) | |
| CN106267897A (en) | Ginsenoside and the method for residual pesticide in sharp separation Radix Ginseng | |
| Shehzad et al. | Rational development of a selection model for solvent gradients in single‐step separation of ginsenosides from P anax ginseng using high‐speed counter‐current chromatography | |
| CN104007219B (en) | A kind of liquid chromatography-enzyme electrode combined apparatus and application thereof | |
| Nováková | Advances in sample preparation for biological fluids | |
| Tang et al. | Screening and isolating potential α-glucosidase inhibitors from Rhizoma Coptidis by ultrafiltration LC-PDA-ESI/MS combined with high-speed countercurrent chromatography and reverse-phase medium-pressure liquid chromatography | |
| Cui et al. | Purification of flavonolignan diastereoisomers from Arenaria kansuensis by two-dimensional liquid chromatography combined with solid-phase extraction | |
| Zhou et al. | Analysis of lignans in Magnoliae Flos by turbulent flow chromatography with online solid‐phase extraction and high‐performance liquid chromatography with tandem mass spectrometry | |
| CN102507794A (en) | Method for rapidly detecting poisoned substances by adopting urine | |
| Sawant et al. | Novel extraction and characterization methods for phytochemicals | |
| CN102507793A (en) | Method for detecting poison quickly by using blood | |
| Rodríguez-Palma et al. | A modified micro-solid phase extraction device for in-port elution and injection into portable liquid chromatography: A proof-of-concept study | |
| Majors | Advanced topics in solid-phase extraction: chemistries | |
| Zhang et al. | Coupling of micro solid-phase extraction with electrospray ionization and its potential for complex sample analyses using a miniature mass spectrometer | |
| CN104897815B (en) | Method for analyzing active ingredients of PCPs (personal care products) in indoor dust | |
| Liu et al. | Quantitative Determination of 7 Saikosaponins in Xiaochaihu Granules Using High‐Performance Liquid Chromatography with Charged Aerosol Detection |
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 |