CN1289922A - Biochemical Process for preparing liquid-phase chromatograph - Google Patents
Biochemical Process for preparing liquid-phase chromatograph Download PDFInfo
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Abstract
A liquid-phase chromatograph for purifying bioengineering products is composed of flow-phase tank, ordinary-temp chromatographic peristaltic pump, middle-pressure chromatographic titanium pump, 10-way valve, Ti-alloy chromatographic column, detector, automatic fraction collector, and chromatographic workstation for controlling data acquisition and processing. Its advantages include small size and low cost.
Description
The present invention relates to the separation equipment in a kind of Bio-engineering Products purifying, be specifically related to a kind of liquid chromatograph for preparing usefulness.
Biological preparative chromatography has been subjected to people's great attention and has obtained widespread use as an important branch of separation science.In bio-pharmaceuticals and bioengineering field, be used for preparative-scale purifying biological engineering product, it has become separation and Extraction albumen, enzyme, antibody and nucleic acid preparation or the essential separation and purification means of other biochemical components.In the Bio-engineering Products purge process, fermentation liquor often need carry out the roughing out of component in the normal pressure chromatogram behind centrifugal, ultrafiltration and concentration, tells to press that chromatogram is smart separates in carrying out after the fraction that needs, in the hope of making specification product again.
At present, the liquid chromatograph of domestic production is analytic type, biochemical preparative chromatography main dependence on import.But, these import equipments mainly are some common apparatus, be exclusively used in the biochemical preparation not really desirable, not high as automaticity, work such as particularly data acquisition, analysis, processing, report and printing, the import equipment control time generally has only 30-40 minute, and in the gene engineering product production technology, purge process often continues tens hours even several days; Simultaneously, be subjected to the long restriction of disengaging time of preparative chromatography, import equipment can't be controlled in real time and handle with microcomputer, can not carry out the collection control that gradient elution control, single-pump flow control and chromatogram flow out component with microcomputer; In addition, the import equipment normal pressure separates needs two complete equipments with middle pressure, not only in use make troubles, and more can increase engineering cost, and the preparative liquid chromatograph of normal pressure, middle pressure coupling is not arranged at present as yet; Also have, import equipment is bulky, and adopts case structure, though comparatively attractive in appearance in appearance, the operation and maintenance trouble.
Purpose of the present invention is to provide a kind of purifying that can be advantageously used in Bio-engineering Products, integrates normal pressure and middle pressure chromatographic resolution, has suitable chromatographic work station control, the biochemical Process for preparing liquid-phase chromatograph that volume is less, preparation cost is lower.
For achieving the above object, technical scheme below adopting:
The invention discloses a kind of biochemical Process for preparing liquid-phase chromatograph, comprise moving phase storage narrow-necked earthen jar, inject the chromatographic work station etc. of pump, valve, chromatographic column, detecting device, gatherer and the control data collection and the processing of moving phase, it is characterized in that, described pump is normal pressure chromatogram peristaltic pump and middle pressure chromatogram titanium pump, described chromatographic column is the titanium alloy chromatographic column, described gatherer is an automatic fraction collector, or ten channel valve collect as press the peak, and described chromatographic work station realization is to the control of pump, valve.
Above-mentioned biochemical Process for preparing liquid-phase chromatograph, described chromatographic work station are the intelligent chromatographic work station of TL-2000; Described sampling valve is seven logical valves; Adopt brick pattern on the structures such as chromatographic work station of pump, valve, chromatographic column, detecting device, gatherer and the control data collection and the processing of described moving phase basin, injection moving phase; Described detecting device list wavelength detects, and wavelength is 280nm, changes optical filter and also can change wavelength.
Adopt above-mentioned design and structure, have following advantage:
1, owing to realize that peristaltic pump and titanium pump switch and use, make an instrument can be successively as the normal pressure chromatogram and
The middle chromatogram of pressing is used, and is economical and practical, thereby can reduce the cost of biochemical preparation;
2, adopt titanium alloy pump and titanium alloy chromatographic column, have good bio-compatibility, make the equipment can be special-purpose
In bioengineering, be applicable to and separate activated biochemical component;
3, select for use suitable also successful operation can carry out the chromatographic work station of external control pump, gradient and gatherer, worker
Do the flow process full-automation;
4, adopt ten passages to collect valve, avoided waiting to collect the component volume restrictions and making whole as the component gatherer
Machine is more coordinated, and volume is littler;
5, single injected sampling can prepare hundreds of milligram to tens gram sample, and is alternative big, satisfies bioengineering
Needs.
Below in conjunction with accompanying drawing in detail the present invention is described in detail:
Fig. 1 is a structural representation of the present invention;
Fig. 2 is a system construction drawing of the present invention;
Fig. 3 examines as light source MP-D type ultraviolet for using electrodeless discharge lamp in the instrument of the present invention
Survey device long term stability tests curve map;
Fig. 4 is for examining as the 8823A type ultraviolet of light source with hollow cathode lamp in the instrument of the present invention
Survey device long term stability tests curve map;
Fig. 5 is chromatographic work station of the present invention system connection layout;
Fig. 6 is that instrument application of the present invention is in separating Chinese cobra snake venom chromatogram;
Fig. 7 is instrument application of the present invention effective constituent chromatogram in separated plasma;
Fig. 8 is that instrument application of the present invention is in separating blood vessel endothelium chalone chromatogram;
Fig. 9 separates the reappearance chromatogram of Chinese cobra snake venom for instrument of the present invention.
Referring to Fig. 1 and Fig. 2, chromatograph of the present invention comprises moving phase storage narrow-necked earthen jar 1, injects the parts such as pump 2, valve 3, chromatographic column 5, detecting device 6, gatherer 7 and chromatographic work station 8 of moving phase, and chromatographic work station 8 is finished data acquisition, processing and to the control of pump, valve.
Pump described in the present invention 2 adopts peristaltic pump during the normal pressure chromatographic resolution, be used to drive liquid by chromatographic column and other liquid road.These peristaltic pump characteristics are:
(1) peristaltic pump is by moving 5-8 the roller bearing of a centre wheel string, and extruding elastic hose when rotating again makes liquid reach the effect of carrying liquid to a direction extruding.Its roller bearing of general peristaltic pump all is passive, i.e. itself not rotation, and frictionally rotates and also pressed flexible pipe fully.Flexible pipe is squeezed and can extends, elongate toward a direction, can rupture because of wearing and tearing at last.The roller bearing of the peristaltic pump of selecting for use among the present invention is initiatively, and when centre wheel rotated, roller bearing press reverse direction and rotated, and flexible pipe only is squeezed and does not stretch.Therefore, prolonged the serviceable life of flexible pipe.The wearing and tearing of flexible pipe and replacing can make flow velocity change, so use this pump can obtain long-time constant flow stream velocity.
(2) convenient for changing, peristaltic pump can be furnished with general flow (0.01 μ l/ branch-30ml/ branch) and two kinds of pump heads of big flow (1ml/ branch-70ml/ branch).General flow pump head has 8 roller bearings, and the high-capacity pump head has 5 roller bearings.Roller bearing is many more, and the pulsation of pumping solution is more little.
(3) peristaltic pump need be furnished with RS-232 interface, is connected to chromatographic work station 8 by this interface, controls it by workstation and opens, stops and flow velocity (method of operating is seen chromatographic work station).
(4) flexible pipe of peristaltic pump is generally silicone rubber tube, has various internal diameters available, but pipe thickness is answered consistent (1.5mm), otherwise can not use.Silicone rubber tube can only tolerate water, methyl alcohol, ethanol and ethylene glycol isopolarity solvent, meets non-polar solvents such as acetone, chloroform, ethyl acetate and benzene and promptly can not be used by swelling.Can use extraordinary flexible material pipes such as Viton, fluororubber or fluorosioloxane rubber this moment instead, or use the silicone rubber tube of liner tetrafluoro.The tearing toughness of silicone rubber tube is low, can not meet sharp instrument and rupture in case cut out wound.Silicone rubber tube can tolerate 110 autoclavings.Can unload when needing sterile working, wrap, load onto use after the sterilization with cloth.
The peristaltic pump that the present invention selects for use, can be with reference to table 1 parameter:
Table 1: flow range when peristaltic pump of the present invention is unloaded
The present invention is during as middle pressure chromatographic resolution, and described pump 2 adopts titanium alloy pumps.This titanium pump should have following characteristics:
(1) flow is the titanium pump of 0.1-40ml/min or 0.1-80ml/min, sees table 2 for details.
(2) parts of titanium pump contact liq are made by materials such as titanium alloy, fluoroplastic and jewels.
(3) titanium pump can be used manual operation, and keyboard starts, and also can be connected with chromatographic work station by RS-232 interface is set, and controls it by workstation and opens, stops and flow velocity (method of operating is seen chromatographic work station).
Titanium pump of the present invention is that the constant volume mode is worked, so should be furnished with pressure demonstration and overvoltage and under-voltage protection function.
Table 2: flow range when titanium pump of the present invention is unloaded
The present invention uses chromatographic column 5 to be stainless steel analytical column, normal pressure glass column and middle pressure chromatogram titanium alloy post and macromolecule material chromatographic column.
The present invention uses gradient valve 3 as 24V DC three tunnel two direction changeover valves, is driven by chromatographic work station.
Simultaneously, use seven logical valves 31 to be used for quantitative sample injection, with injection annulus or manage 32 and is connected, between pump 2 and post 5, be provided with and adorn sample, sample introduction and three positions of washing.
The eluent that flows out from chromatographic column 5 among the present invention has carried post and has separated eluted each composition in back, and the UV-detector 6 of flowing through one by one produces absorption.Just constitute the chromatogram Figure 81 that forms by a series of chromatographic peaks with effluent volume and optical density mapping.
UV-detector 6 can be selected two kinds, promptly with electrodeless discharge lamp as the MP-D type UV-detector of light source with the 8823A type UV-detector of hollow cathode lamp as light source.These two kinds of UV-detector long-time stability are tested: be placed in the chromatography refrigerator, constant temperature is in 4 ~ 8 ℃, in detection cell, be full of 2% isopropyl alcohol, open uviol lamp, signal output part is connected to central detection system, simultaneously the temperature variation of two positions about detection system, room temperature and the chromatography refrigerator, test duration is 00,000 minute, write down its situation of change, see Fig. 3, Fig. 4.
According to test result, the temperature in the chromatography refrigerator does not influence the stability of UV-detector substantially; From the long term drift situation, the MP-D long-time stability are better than the 8823A type slightly, are applicable to the chromatographic resolution that the working time is long; And after lighting use for a long time, 8823A type stability makes moderate progress, because its wavelength replacing is comparatively convenient, applicable to short chromatographic resolution of working time.
When light source is selected the electrodless discharge iodine lamp for use, select monochromatic light from the top bright dipping by interference filter, wavelength is generally used 280nm, can choose 260nm, 206nm etc.Protein isolate is generally used 280nm, also can obtain higher sensitivity (be about the light absorption of 280nm place 50 times) at the 206nm place.
This detecting device can use in the chromatography refrigerator for a long time among the present invention, and the build-up of luminance of turning on light when being preferably in the outer room temperature of cabinet moves into refrigerator with a whole set of instrument then, just no longer turns off the light in case move into refrigerator.Lamp life-span＞10,000 hour, (being not related to round the clock) use was more than 1 year continuously.If have the long period need not, then a whole set of instrument can be moved to from refrigerator under the room temperature, placing after 2 hours outage again and turning off the light, in order to avoid there is cold air to coagulate in circuit.This detecting device can manually use (joining registering instrument), also can with the chromatographic work station coupling.
Gatherer 7 among the present invention, can use Beijing new technology application research or the automatic fraction collector produced of Shanghai Xin Bo radio factory, this gatherer has regularly, meter drips a function, is used in the collection of component in a small amount; Can also be equipped with 1 * 10 more advanced valve and collect usefulness as pressing the peak.Gatherer and chromatographic work station coupling.
The chromatographic work station that the present invention adopts, be the commercial chromatographic work station of the TL-2000 type of Thailand uprightization Electron Technology Co., Ltd exploitation, this chromatographic work station is applicable to the needs of preparative liquid chromatograph of the present invention, except that having data acquisition and processing capacity, also have control pump, valve formula gradient, double pump inside gradient and by functions such as peak collections.
Above-mentioned chromatographic work station is nursed one's health control module, main frame, three major parts of workstation software by intelligence and is formed, and sees also accompanying drawing 2,5.
The TL-CSITU-8 type intelligence conditioning unit of intelligence conditioning control module (being called for short the regulation and control unit) safe uprightization of employing Electron Technology Co., Ltd.It is made up of control module 82 and conditioning unit 83.Control module 82 is used for to the control of gradient elution solenoid valve 3, unit flow pump 2 and component gatherer 7; Conditioning unit 83 is used for converting the peak-to-peak signal (simulating signal) of chromatographic detector 6 outputs to digital signal that computing machine can be discerned;
Main frame 84 is the carrier of workstation;
Workstation software is the intelligent chromatographic work station system software of the safe uprightization exploitation TL-2000 of Electron Technology Co., Ltd type, and the control of finishing control module is reached identification, processing, integration, calculating to peak-to-peak signal, finally obtain the experimenter need data result.
Workstation software compiles according to following major control flow process:
For finishing the present invention, need by the part operation chromatographic work station that cooks noodle:
Prepare chromatograph (CE or biochemical preparative chromatograph can be controlled four cover chromatographic systems at most simultaneously for GC, LC); One of microcomputer: 80586 (or higher) processor need be arranged, internal memory 32M, the above idle hard disk of 120M, SVGA display (display resolution 800 * 600), CPU frequency 100M, Win95 or Win98 operating platform, one of QWERTY keyboard, one of Genius mouse, one of printer; A computing machine can be joined four intelligence conditioning control modules at most, controls the quadruplet chromatographic system simultaneously.
The installation of chromatographic work station:
The installation of chromatographic work station is divided into hardware and software two parts:
Chromatographic work station of the present invention is mainly by connecting to form system hardware with the lower part:
1 in main frame, 1 of display, 1 on keyboard, 1 of mouse, 1 of printer, 6 of floppy disks (workstation software), chromatogram intelligence conditioning control module is looked demand configuration (1-4 platform), cable, signal wire, data line, teleswitch, 1 in every passage, 1 of softdog.
Wherein, conditioning unit rear panel is established:
RS-232: standard digital communication serial ports, by data line and corresponding host communication mouth (com) connection.RS-232 interface is a kind of interface that passes elm data and control command between conditioning unit and the chromatographic work station main frame.Its shape can be for having the rectangle in 25 cushion holes.
The a plurality of conditionings of hyperchannel workstation configuration unit.The RS-232 communication port of each conditioning unit connects a communication port (com) of workstation main frame by data line.
REMOTE: remote control starting switch jack.
When the workstation main frame was far away apart from chromatograph, for guaranteeing retention time comparatively accurately, inconvenience started the workstation sampling routine with mouse and keyboard behind the sample introduction, in this case, can start sampling with teleswitch, and method is the button that clicks the switch top.
The shape of REMOTE jack can be the circle hole shape that has five pins.
INPUT: input end of analog signal.This terminal is connected with the chromatograph signal output part by signal wire, accepts the analog voltage signal of chromatographic detector output.
The shape of INPUT terminal can be the circle hole shape that has five pins.
FUSE: protective tube, 0.2 ampere of ratings.
AC-LINE: AC Power supply socket connects the alternating current 220V power supply by power lead.
Wherein nursing one's health the unit front panel establishes:
POWER: power switch.
Display screen: the electric signal that shows chromatograph output with the eight digit number code tracking.The digital is-symbol position of Far Left that shows, polarity "+[just] " or " [the bearing] " of expression chromatogram electric signal.Middle six decimal number word bits show chromatographic signal instantaneous voltage value, and the unit millivolt upgrades with per second speed once.One of rightmost is the range position.
ZERO: zero-setting button, its function is: press this button, workstation promptly with current magnitude of voltage as relative zero.
This button generally is used for following several situation:
When the chromatograph output signal can not be continued furnishing zero again, its baseline polarization or negative partially, and the operator wishes this baseline value as reference zero point (relative zero), later signal is during as benchmark, can click this zero-setting button, the conditioning unit can be automatically with current demand signal as reference zero point, digitally show vanishing.
The zeroing of conditioning unit self calibrating.Because long-term use, season or variation of ambient temperature, zero point drift also can take place in conditioning unit itself, at this moment also can carry out zero point if desired and adjust.
When doing high accuracy analysis, for example do the high duplication experiment, each sample analysis time surpasses one hour, and relative drift may take place baseline before and after the experiment, adjusts if carry out zero point before each sample introduction, can guarantee the high precision of analytical structure.
F: range selection button, pressing this button continuously can switching range, digital last position that shows of display screen can show successively " H " (20mV～+ 1180mV), " L ", (10mV～+ 140mV)
“E”(-1．25mV～+1．25mV)。
Wherein, the control module rear panel mainly comprises 24 volts of tributary power inputs (solenoid valve), solenoid valve binding post, component gatherer binding post, component gatherer power input, communication port (controlling two peristaltic pumps), PORT COM (controlling two titanium pumps).
The control module front panel mainly comprises:
Rs-232 standard digital communication serial ports is by data line and the corresponding communication port connection that is subjected to sky parts (as pump and gatherer).Rs-232 standard digital communication serial ports is a kind of interface that transmits data and control command between control module and the chromatographic work station main frame, and its shape can be for having the rectangle in 5 cushion holes.
Also comprise control module power switch and control module supply socket
With conditioning recited above unit and control module, and mouse, printer and display and main frame couple together.Softdog directly is inserted on the LPT, print cable is inserted in the other end of softdog again.
System connect finish check errorless after, according to the software document install software, the debugging software and hardware carries out separation test.
Use the good chromatograph of the present invention of above-mentioned Installation and Debugging, carry out following biological sample discrete testing:
One, the Chinese cobra snake venom separates
Contain 6% α-Cobratoxin in the Chinese cobra snake venom, this is a kind of peptide constituents that very strong analgesic effect is arranged, molecular weight 6960.Might oral administration, the threshold of pain obviously raises during rat dosage 1.25-5 μ g/kg, and analgesic activity is stronger than morphine, and duration of efficacy is also long than morphine, does not have a habituation.
Ion-exchange chromatography separate mode: Triacryl-CM is stationary phase (5ml), and ammonium acetate is a moving phase, valve formula gradient (0.05M, pH5.0～0.5M, pH7.0; 0.5M, pH7.0～1M, pH7.0), applied sample amount 50mg (2ml), flow velocity 1.0ml/min, λ=280nm.
Separating resulting is referring to Fig. 6.A chromatographic resolution 50 milligrams of snake venom, the peak that wherein indicates " ↓ " is the Cobratoxin peak, repeatedly repeated isolation all obtains satisfactory result, repeatability fine.
Two, the separation of effective constituent in the blood product
Contain many useful components in the human plasma, Antithrombin III is a kind of anticoagulant protein in the human plasma, and it can be used for bypass surgery and make anti-coagulants, and no antigen is nontoxic, and content is per hundred milliliters 29 milligrams in human plasma.
The affinity chromatography separate mode: heparin is stationary phase (4ml), and Tris-HCL is a moving phase, valve formula gradient, applied sample amount 75ml, flow velocity 7.0 ml/min, λ=280nm.
Separating resulting has the peak of arrow to represent to have the peak of anticoagulating active referring to Fig. 7, just can obtain more Antithrombin III as long as amplify chromatographic column.Minute can finish surplus in the of 20 successively and separate, obtain satisfied separating resulting.
Three, the separation of bioengineering product-blood vessel endothelium chalone
The blood vessel endothelium chalone of genetic recombination is a kind of recombinant protein of molecular weight about 30000, can stop tumor tissues microvascular endothelial growth on every side, suppresses tumor growth and disperse thereby reach.
The affinity chromatography separate mode: heparin is stationary phase (25ml), and Tris-HCL is a moving phase, valve formula gradient, applied sample amount 10mg (15ml), flow velocity 1.0ml/min, λ=218nm.
Separation chromatography obtains satisfied separating resulting referring to Fig. 8.
Four, utilize chromatograph of the present invention to separate the reproducible mensuration of Chinese cobra snake venom:
With the Chinese cobra snake venom is sample, and difference sample introduction 50mg and 40mg separate with identical condition of gradient elution on 30 * 20mm Triacryl-CM chromatographic column, and the record separating resulting is seen Fig. 9.
From test result as can be seen, preparative liquid chromatograph of the present invention can obtain reappearance separating resulting preferably when the separate complex protein component, can be applied to the preparation separation and purification of actual sample.
Five, preparative liquid chromatograph recovery experiment of the present invention
1. experiment material:
Sample: 1%IgG (people) normal saline solution;
Eluent A:0.01mol/L Tris-HCL, pH 7.4;
Eluent B:0.0lmol/L Tris-HCL+0.05mol/L NaCl, pH7.4;
Chromatographic column: the DEAE-Trisacryl ion exchange column (20 * 32mm, 5ml);
1 * 5ml volumetric flask, 5 * 10ml volumetric flask, 1 * 100ml volumetric flask, 500 μ l quantitative sample injection pipes, 10ml syringe;
Biochemical Process for preparing liquid-phase chromatograph of the present invention, Spectrumlab 22 type ultraviolet-visible pectrophotometers.
2. experiment condition:
Sample introduction: 500 μ l; Detect wavelength: 280nm; Flow velocity: 1.2ml/min; Parallel five tests.
Valve formula gradient elution:
| Time (min) | B％ |
| 0．0 | 0 |
| 19．0 | 100 |
| 19．2 | 100 |
| 20．5 | 100 |
| 21．0 | 0 |
| 25．0 | 0 |
3. experimental result:
Flow out component with 10.00ml volumetric flask quantitative collection chromatographic column, be equipped with and survey.Parallel five tests are collected with five 10.00ml volumetric flasks respectively.
At Speictrumlab 22 type ultraviolet-visible pectrophotometer 280nm places, as blank absorbent solution, measure the absorbance A that chromatographic column that the 10.00ml volumetric flask collects flows out component respectively with the mixed liquor (respectively accounting for 50%) of eluent A and eluent B
i(I=1,2,3,4,5).
Get 1%IgG normal saline solution 5ml and also be settled in the 100ml volumetric flask, measure its absorbance A as stated above with blank absorbent solution dilution
s
The calculating of the recovery (R):
Because, A=abc; So R=A
i/ A
s
Result of calculation sees the following form.Table: recovery test result
As seen from the above table, average recovery rate is 101.46%.
From above test, as can be seen, preparative chromatograph provided by the invention, separating effect is obvious, and the recovery is higher, and in the operating process, the chromatogram station is working properly.
Claims (5)
1, a kind of biochemical Process for preparing liquid-phase chromatograph, comprise the moving phase basin, inject moving phase pump, valve, chromatographic column,
The chromatographic work stations of detecting device, gatherer and control data collection and processing etc. is characterized in that institute
Stating pump is normal pressure chromatogram peristaltic pump and middle pressure chromatogram titanium pump, and described chromatographic column is the titanium alloy chromatographic column, and is described
Gatherer is an automatic fraction collector, or ten channel valve are collected described chromatographic work station realization as pressing the peak
Control to pump, valve.
2, biochemical system according to claim 1 is put forth energy liquid chromatograph, it is characterized in that described chromatographic work station
Be the intelligent chromatographic work station of TL-2000.
3, biochemical Process for preparing liquid-phase chromatograph according to claim 1 is characterized in that, described sampling valve is seven
Logical valve.
4, biochemical Process for preparing liquid-phase chromatograph according to claim 1 is characterized in that, described moving phase basin,
Pump, valve, chromatographic column, detecting device, gatherer and the control data collection and the processing of injection moving phase
Adopt brick pattern on the structures such as chromatographic work station.
5, biochemical Process for preparing liquid-phase chromatograph according to claim 1 is characterized in that, described detecting device list ripple
The long detection, wavelength is 280nm, changes optical filter and also can change wavelength.
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|---|---|---|---|
| CN 00133303 CN1119654C (en) | 2000-11-21 | 2000-11-21 | Biochemical process for preparing liquid-phase chromatograph |
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|---|---|---|---|
| CN 00133303 CN1119654C (en) | 2000-11-21 | 2000-11-21 | Biochemical process for preparing liquid-phase chromatograph |
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| CN1119654C CN1119654C (en) | 2003-08-27 |
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| CN101747407A (en) * | 2008-12-02 | 2010-06-23 | 杭州中肽生化有限公司 | High-efficient integrated system for separation and purification |
| CN102124329A (en) * | 2008-08-19 | 2011-07-13 | 株式会社岛津制作所 | Split liquid chromatographic apparatus |
| CN101294975B (en) * | 2008-06-19 | 2013-06-05 | 中国科学院生态环境研究中心 | Grouping toxicology analyzer |
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| CN102124329B (en) * | 2008-08-19 | 2014-07-30 | 株式会社岛津制作所 | Split liquid chromatographic apparatus |
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| CN104749309B (en) * | 2015-03-30 | 2018-11-13 | 中国科学院青岛生物能源与过程研究所 | It is a kind of to prepare ion chromatograph |
| CN105374265A (en) * | 2015-12-15 | 2016-03-02 | 徐州医学院 | High efficiency liquid chromatograph experiment teaching model and experiment method |
| CN105374265B (en) * | 2015-12-15 | 2018-05-22 | 徐州医学院 | A kind of high performance liquid chromatograph experimental teaching model of cobalt and experimental method |
| CN106731006A (en) * | 2016-12-07 | 2017-05-31 | 中国农业科学院茶叶研究所 | The method for separating and preparing of one Lepidopterous epoxy olefins isomer |
| CN108459120A (en) * | 2018-01-29 | 2018-08-28 | 天津博纳艾杰尔科技有限公司 | A kind of purifying preparing chromatography system |
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