CA2346358A1 - Method and device for the rapid liquid chromatographic separation of substance mixtures and for the identification of substances - Google Patents
Method and device for the rapid liquid chromatographic separation of substance mixtures and for the identification of substances Download PDFInfo
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- CA2346358A1 CA2346358A1 CA002346358A CA2346358A CA2346358A1 CA 2346358 A1 CA2346358 A1 CA 2346358A1 CA 002346358 A CA002346358 A CA 002346358A CA 2346358 A CA2346358 A CA 2346358A CA 2346358 A1 CA2346358 A1 CA 2346358A1
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- 239000000126 substance Substances 0.000 title claims abstract description 53
- 239000000203 mixture Substances 0.000 title claims abstract description 29
- 238000000034 method Methods 0.000 title claims abstract description 18
- 238000013375 chromatographic separation Methods 0.000 title claims abstract description 9
- 239000007788 liquid Substances 0.000 title abstract description 5
- 238000000926 separation method Methods 0.000 claims abstract description 95
- 238000011144 upstream manufacturing Methods 0.000 claims description 2
- 230000000274 adsorptive effect Effects 0.000 claims 1
- 230000003993 interaction Effects 0.000 claims 1
- 238000000746 purification Methods 0.000 claims 1
- 238000012360 testing method Methods 0.000 abstract description 11
- 230000000694 effects Effects 0.000 abstract description 8
- 238000002955 isolation Methods 0.000 abstract description 5
- 238000007445 Chromatographic isolation Methods 0.000 abstract description 2
- 238000001179 sorption measurement Methods 0.000 description 8
- 239000013543 active substance Substances 0.000 description 7
- 239000000306 component Substances 0.000 description 7
- 238000011067 equilibration Methods 0.000 description 7
- 239000000523 sample Substances 0.000 description 7
- 239000002904 solvent Substances 0.000 description 5
- 239000002699 waste material Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 239000003463 adsorbent Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 239000003480 eluent Substances 0.000 description 3
- 239000000727 fraction Substances 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 2
- 238000004566 IR spectroscopy Methods 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 230000003334 potential effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000009877 rendering Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 238000013022 venting Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/38—Flow patterns
- G01N30/46—Flow patterns using more than one column
- G01N30/461—Flow patterns using more than one column with serial coupling of separation columns
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/18—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns
- B01D15/1864—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns using two or more columns
- B01D15/1871—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns using two or more columns placed in series
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/18—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns
- B01D15/1864—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns using two or more columns
- B01D15/1885—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns using two or more columns placed in parallel
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/16—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the conditioning of the fluid carrier
- B01D15/163—Pressure or speed conditioning
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8804—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 automated systems
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/38—Flow patterns
- G01N30/46—Flow patterns using more than one column
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/38—Flow patterns
- G01N30/46—Flow patterns using more than one column
- G01N30/466—Flow patterns using more than one column with separation columns in parallel
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/26—Conditioning of the fluid carrier; Flow patterns
- G01N30/38—Flow patterns
- G01N30/46—Flow patterns using more than one column
- G01N30/468—Flow patterns using more than one column involving switching between different column configurations
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- Chemical & Material Sciences (AREA)
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- Chemical Kinetics & Catalysis (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
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- Pathology (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention relates to a method and device for the rapid liquid chromatographic separation of substance mixtures and for the identification of substances. The aim of the invention is to provide a device and a method for the liquid chromatographic separation, isolation, and identification of substances in analytic and semipreparative areas, with which a test for determining the activity of substance mixtures is dispensed with. In additio n, the inventive method and device carry out the separation of substance mixtures, and the isolation and identification of the individual substances more quickly than prior art methods and devices. To these ends, a method and device are used with which substance mixtures, in a software-controlled rapi d liquid chromatographic two-step separation, are subjected to a preliminary separation in a first step and, in the second step, the fractions which were subjected to the preliminary separation and which are deposited in collectio n columns are parallelly separated into at least two separation lines in a fin e manner. The finely separated fractions are parallelly identified and parallelly isolated.
Description
Method and Device for the Rapid Liquid Chromatographic Separation of Substance Mixtures and for the Identification of Substances Specification The invention relates to a process and a device far the rapid liquid-chromatographic separation of mixtures of sub-stances and identification of substances according t:o the preambles of claims 1 and 5.
For example, the pharrnaceutical research frequently encoun-ters the problem of isolating pharmaceutically active sub-stances from mixture~~ of substances. Thus, extracts of natural products or mixtures of substances produced by com-binatorial chemistry a:re being rested for potential activ-ity. Using mixtures of: substances found to have activity, attempts are then made to isolate the active substances by means of complex sepax-ation procedures. Thereafter, the in-dividual substances of the mixture isolated in this way are subjected to an activity test once more. The active indi-vidual substances being found are investigated for their structure so as to exclude active substances which might be already well-known. On.e drawback in this process is that when testing mixtures of substances, the activity of indi-vidual substances may be suppressed by superposition ef-fects, leaving these substances undetected. Another draw-back is possible simulation of activity by superposition effects, followed by a cost-intensive and useless search for these supposed active substances in the mixture of_ sub-stances. Ultimately, it is disadvantageous that substances already well-known are. excluded only after performing at least two tests for biological activity and after complex isolation procedures, which is very costly. As a rule, large quantities of material are required to perform these tests, i.e., separations have to be carried out on a prepa-rative scale. With respect to capital investment, however, preparative units are more expensive compared to analytical units. Also, preparative units use considerably higher amounts of solvents and buffer substances during separa-tion, rendering their operation costly and, in addition, giving rise to major problems with disposal and to environ-mental pollution.
The invention is based on the obj ect of providing a device and a process for tree liquid-chromatographic separation, isolation and identification of substances in an analytical and semi-preparative :range, by means of which testing for activity of mixtures of: substances is rendered unnecessary, and which enable separating mixtures of substances and iso-lating and identifying the individual substances more rap-idly than is possible up to now.
Said object is accomplished through the characterizing parts of claims 1 and 5.
Advantageous developments are specified in the subclaims.
The invention has various advantages. Double testing of substances, namely, beforehand in the mixture of substances and after isolation, is no longer necessary. According to the invention, the complex and costly, and in part faulty first activity test of the mixtures of substances can be omitted. Instead, fol7_owing combined isolation and identi-fication, only potentially new active substances are sub-jected to further tests. One can do without costly treat-ment of substances already well-known, as is common prac-tice up to now. The input of time and cost for detecting a new active substance c:an be reduced substantially. In addi-tion, this procedure is more reliable because the test re-sults on unknown individual substances are unequivocal and also, all of the active substances present in the mixture will be detected.
The mixtures of substances to be investigated are processed in a two-stage separation wherein, because the separating columns and solid-pha:~e extraction columns (collecting col-umns) are connected with the pump unit in a fashion accord-ing to the invention, parallel separation of multiple frac-tions from the first separation step is possible in the second chromatographic: separation stage. Consequently, this apparatus works substantially more rapid and thus, more economical as compared. to well-known two-stage apparatus.
The individual substances are identified using per se known direct computer-controlled comparison of chromatograms and spectra obtained from detectors, and of the retention range from the first separation step and the retention time from the second separation step with information on well-known substances in a data. base. Ultraviolet absorption, mass spectrometry, light scattering, fluorescence, infrared spectroscopy, and nuclear magnetic resonance spectroscopy are possible as principles of detection and identification.
It is also possible to use additional identificatian pa-rameters such as source and origin of the sample. Because a smaller number of tests are required to identify the sub-stances in the mixture and exclude substances already well-known, the plant can be dimensioned for analytical and semi-preparative scales. Analytical and semi-preparative units are much more economical with respect to initial cost and operation compared to preparative units commonly used so far. Owing to the reduced consumption of solvent and buffer substances, the process and device according to the invention are environmentally beneficial as a result of re-duced amounts of waste.
The invention will be :illustrated in more detail with ref-erence to the drawings and embodiments:
Fig. 1 is a schematic: representation of the operating se-quence of equilibrating in the first separation step and washing of the feed column battery;
Fig. 2 is a schematic; representation of the separation of a mixture of substances in the first separation step and the .adsorption of fractions on the first collecting column battery;
Fig. 3 is a schematic' representation of the separation of a mixture of substances in the first separation step and the adsorption of fractions on the second collecting column battery;
Fig. 4 is a schematic: representation of the separation of a mixture of substances in the first separation step and the ,adsorption of fractions on the third collecting column battery;
Fig. 5 is a schematic: representation of the equilibration of the separation column batteries of the second separation step;
Fig. 6 is a schematic representation of a parallel separa-tion of adsorbed fractions in the second separation step; and Fig. 7 is a schematic: representation of the equilibration of a collecting column battery.
For example, the pharrnaceutical research frequently encoun-ters the problem of isolating pharmaceutically active sub-stances from mixture~~ of substances. Thus, extracts of natural products or mixtures of substances produced by com-binatorial chemistry a:re being rested for potential activ-ity. Using mixtures of: substances found to have activity, attempts are then made to isolate the active substances by means of complex sepax-ation procedures. Thereafter, the in-dividual substances of the mixture isolated in this way are subjected to an activity test once more. The active indi-vidual substances being found are investigated for their structure so as to exclude active substances which might be already well-known. On.e drawback in this process is that when testing mixtures of substances, the activity of indi-vidual substances may be suppressed by superposition ef-fects, leaving these substances undetected. Another draw-back is possible simulation of activity by superposition effects, followed by a cost-intensive and useless search for these supposed active substances in the mixture of_ sub-stances. Ultimately, it is disadvantageous that substances already well-known are. excluded only after performing at least two tests for biological activity and after complex isolation procedures, which is very costly. As a rule, large quantities of material are required to perform these tests, i.e., separations have to be carried out on a prepa-rative scale. With respect to capital investment, however, preparative units are more expensive compared to analytical units. Also, preparative units use considerably higher amounts of solvents and buffer substances during separa-tion, rendering their operation costly and, in addition, giving rise to major problems with disposal and to environ-mental pollution.
The invention is based on the obj ect of providing a device and a process for tree liquid-chromatographic separation, isolation and identification of substances in an analytical and semi-preparative :range, by means of which testing for activity of mixtures of: substances is rendered unnecessary, and which enable separating mixtures of substances and iso-lating and identifying the individual substances more rap-idly than is possible up to now.
Said object is accomplished through the characterizing parts of claims 1 and 5.
Advantageous developments are specified in the subclaims.
The invention has various advantages. Double testing of substances, namely, beforehand in the mixture of substances and after isolation, is no longer necessary. According to the invention, the complex and costly, and in part faulty first activity test of the mixtures of substances can be omitted. Instead, fol7_owing combined isolation and identi-fication, only potentially new active substances are sub-jected to further tests. One can do without costly treat-ment of substances already well-known, as is common prac-tice up to now. The input of time and cost for detecting a new active substance c:an be reduced substantially. In addi-tion, this procedure is more reliable because the test re-sults on unknown individual substances are unequivocal and also, all of the active substances present in the mixture will be detected.
The mixtures of substances to be investigated are processed in a two-stage separation wherein, because the separating columns and solid-pha:~e extraction columns (collecting col-umns) are connected with the pump unit in a fashion accord-ing to the invention, parallel separation of multiple frac-tions from the first separation step is possible in the second chromatographic: separation stage. Consequently, this apparatus works substantially more rapid and thus, more economical as compared. to well-known two-stage apparatus.
The individual substances are identified using per se known direct computer-controlled comparison of chromatograms and spectra obtained from detectors, and of the retention range from the first separation step and the retention time from the second separation step with information on well-known substances in a data. base. Ultraviolet absorption, mass spectrometry, light scattering, fluorescence, infrared spectroscopy, and nuclear magnetic resonance spectroscopy are possible as principles of detection and identification.
It is also possible to use additional identificatian pa-rameters such as source and origin of the sample. Because a smaller number of tests are required to identify the sub-stances in the mixture and exclude substances already well-known, the plant can be dimensioned for analytical and semi-preparative scales. Analytical and semi-preparative units are much more economical with respect to initial cost and operation compared to preparative units commonly used so far. Owing to the reduced consumption of solvent and buffer substances, the process and device according to the invention are environmentally beneficial as a result of re-duced amounts of waste.
The invention will be :illustrated in more detail with ref-erence to the drawings and embodiments:
Fig. 1 is a schematic: representation of the operating se-quence of equilibrating in the first separation step and washing of the feed column battery;
Fig. 2 is a schematic; representation of the separation of a mixture of substances in the first separation step and the .adsorption of fractions on the first collecting column battery;
Fig. 3 is a schematic' representation of the separation of a mixture of substances in the first separation step and the adsorption of fractions on the second collecting column battery;
Fig. 4 is a schematic: representation of the separation of a mixture of substances in the first separation step and the ,adsorption of fractions on the third collecting column battery;
Fig. 5 is a schematic: representation of the equilibration of the separation column batteries of the second separation step;
Fig. 6 is a schematic representation of a parallel separa-tion of adsorbed fractions in the second separation step; and Fig. 7 is a schematic: representation of the equilibration of a collecting column battery.
Fig. 1 through Fig. 7 representatively illustrate the con-figuration and flow scheme of a device according to the in-vention, including a ~~eparation column and three downstream separation lines.
A pump unit. 2 comprised of three pumps 2.1 through 2.3 is connected via 6-way 2-position valves 3.1 and 3.3 and 3-way 2-position 'valve 5.7 l..a a feed column battery 6, a separa-tion column 10 for the first separation stage, and a second separation stage comprised of three separation lines oper-able in parallel, each of which having an upstream 6-way 2-position valve :3.5, 3.6 and 3.7, respectively. In this way, it is possible to convey the mobile phase in any desired composition to any region of the device in a consecutive as well as a parallel fashion.
Each separation line h.as a collecting column battery 7, 8 and 9 and a separation column battery 11, 12 and 13. Repre-sentatively, the collecting column battery 7 includes the collecting columns 7.1. through 7.6, and the separation col-umn battery 11 includes the separation columns 11.1 and 11.2. The other two illustrated separation lines are iden-tical in configuration. Other variants including more feed columns 6.1 through 6.6 in the feed column battery 6, mul-tiple separation columns 10, more than three collecting column batteries 7, 8 and 9, each having more than six col-lecting columns, and more than three separation column bat-teries il, 12 and 13 having more than six separation col-umns per battery are also possible.
The operating sequence of the process according to the in-vention will be described in an exemplary fashion below.
Samples of mixtures of substances are dissolved in a sol-vent and added with an adsorbent each time. Subsequently, the solvent is removed using a rotary evaporator so that the adsorbent covered with sample material achieves flow-ability properties. Th.e adsorbents covered with the mixture of substances are fil7_ed into the feed columns 6.1 through 6.6 of feed column battery 6 and secured in feed column battery 6. The subsequent program sequence steps are con-trolled by a software.
The separation column 10 is equilibrated according to Fig.
1. In a parallel operation, the air is removed from the feed column battery 6. Via pump 2.3, 3-way 2-position valve 5.7 and 6-way 2-position valves 3.1 and 3.3, the air is re-moved with water from one of the dry-filled feed columns 6.1 through 6.6 to be injected next. At the same time, the separation column 10 is equilibrated with a suitable mo-bile solvent via pump 2.1 and 6-way 2-position valves 3.1 and 3.3.
Fig. 2 illustrates the separation of the mixture of sub-stances in the first separation stage on separation column and the subsequent adsorption of the fractions in a separation line inc:Luding the collecting columns 7.1 through 7.6 of collecting column battery 7.
Once the air has been removed from one of the feed columns 6.1 through 6.6, the separation program is started. Ini-tially, the 6-way ~;-position valves 3.3 and 3.5 are switched in position. Via a low-pressure valve unit 1 in-cluding the low-pressure valves 1.1 through 1.3, the compo-nents of the mobile ~ohase can be fed into the system by means of pump unit 2. The mobile phase is conveyed via the low-pressure valve 1..1 of pump 2.1 and via pump 2.1, and this system can be run both in an isocratic fashion and with a gradient. Via 6-way 2-position valve 3.3 and 7-way 6-position valves 4.1; ~4.2, the mobile phase is conveyed by pump 2.1 to that particular feed column 6.1 through 6.6 from which sample material is to be processed. The sample to be separated is transferred from one of the feed columns _ 7 -6.1 through 6.6 to the separation column 10. Via a 6-way 2-position valve 3.4 and. detector 14.1, the separated sample components discharging from separation column 10 arrive at T-piece 17, where water is mixed into the mobile phase via pump 2.2 and 6-way 2-position valve 3.1. The amount of ad-mixed water depends on the polarity of the substances to be separated. Now, as the polarity of the mobile is increased by the water, adsorption on collecting columns 7.1 through 7.6 of collecting column battery 7 is possible. Initially, adsorption is effected on the collecting column battery 7 via 6-way 2-position valve 3.5, the collecting columns 7.1 through 7.6 being loadE=_d with fractions one by one.
Fig. 3 illustrates the adsorption of additional fractions on the collecting co7_umns 8.1 through 8.6 of collecting column battery 8. Once all of the collecting columns of collecting column battery 7 are loaded with fractions, the 6-way 2-position valves 3.5 and 3.6 switch the collecting column battery 8 into the eluent stream. Now, the collect-ing columns 8.1 through 8.6 are loaded with fractions one by one.
Fig. 4 illustrates the adsorption of fractions on the col-lecting columns 9.1 through 9.6 of collecting column bat-tery 9. Once all of the collecting columns 8.1 through 8.6 of collecting column ibattery 8 are loaded with fractions, the 6-way 2-position valves 3.6 and 3.7 switch the collect-ing column battery 9 ~_nto the eluent stream. Now, the col-lecting columns 9.1 through 9.6 are loaded with fractions one by one. In the next. operating sequence step, the frac-tions adsorbed on the three collecting column batteries 7, 8 and 9 are eluted in parallel and separated further on the appropriately assigned separation column batteries 11, 12 and 13.
The separation column batteries 11, 12 and 13 are equili-brated prior to each separation. Fig. 5 illustrates the equilibration of separation column batteries 11, 12 and 13.
For equilibration, the mobile phase is conveyed to separa-tion columns 11.1 and 11.2 of separation column battery 11 via pump 2.1 and 6-waxy 2-position valves 3.1 and 3.5, re-spectively. From there, the mobile phase is conveyed to the waste via 6-way 2-posit: ion valve 3.4, detector 14.1, and a fraction collector 15.1. In a parallel operation, the sepa-ration columns 12.1 and 12.2 of the separation column bat-tery are equilibrated via pump 2.2, the 6-way 2-position valves 3.1 and 3.6, a detector 14.2, and fraction collector 15.2. Likewise, the separation columns 13.1 and 13.2 are equilibrated via pump 2.3, the 6-way 2-position valve 3.7 and 3-way 2-position valve 5.7, a detector 14.3, and a fraction collector 15.3 i.n a parallel operation.
Fig. 6 illustrates the parallel separation of fractions ad-sorbed on collecting column batteries 7, 8 and 9, using the separation column batteries 11, 12 and 13. To initiate the separation step, the mobile phase is conveyed to the col-lecting column battery 7 via pump 2.1 of pump unit 2 and 6-way 2-position valves 3.1 and 3.5. The first eluted frac-tion from collecting column battery 7 (e. g. from collecting column 7.1) is passed to separation column battery 11 via 6-way 2-position valve 3.5. There, one of the separation columns 11.1 or 11.2 c:an be switched in, optionally in a software-controlled f<~shion. Subsequently, the separated components are passed to the detector 14.1 via 6-way 2-po-sition valves 3.5 and 3.4. The software in the electronic control unit assesses the signals by peak detection, di-recting the separated components into the appropriate vials of fraction collector 15.1. Simultaneously, time control of the fraction collector 15.1 is also possible. Such time control can be activated automatically in case no peak passes the detector.
- g _ In a parallel operation, the mobile phase is conveyed to the collecting columr.~ battery 8 via pump 2.2 and 6-way 2-position valves 3.1 and 3.6. The first eluted fraction from collecting column. battery 8 (e. g. from collecting col-umn 8.1) is passed to separation column battery 12 via 6-way 2-position valve 3.6. There, one of the separation col-umns 12.1 or 12.2 can be switched in, optionally in a soft-ware-controlled fashion. The separated components are passed to the detector 14.2. In this case as well, the software assesses the signals by peak detection, subse-quently directing the separated components into the appro-priate vials of fraction collector 15.2. Similarly, the fraction collector 15.2 can be operated in a time-con-trolled fashion. Such time control can be activated auto-matically in case no peak passes the detector.
In parallel to the operations in two separation lines, the third separation line is activated with respect to initiat-ing the separation step. To this end, the mobile phase is passed to collecting column battery 9 via pump 2.3 and 3-way 2-position valve 5.7 and 6-way 2-position valve 3.7.
The first eluted fraci=ion from collecting column battery 9 (e. g. from collecting column 9.1) is passed to separation column battery 13 via valve 3.7. There, one of the separa-tion columns 13.1 or 1.3.2 can be switched in, optionally in a software-controlled fashion. The separated components are passed to the detector 14.3. The downstream fraction col-lector 15.3 is controlled as described above. After each one of the first fractions has been processed in parallel, the separation columzi batteries 11, 12 and 13 are re-equilibrated (cf . , FicT,. 5) for preparation and in order to separate the next fractions. Subsequently, the 7-way 6-po-sition valves 4.3/4.4, 4.5/4.6 and 4.7/4.8 on the collect-ing column batteries 7, 8 and 9 are switched over, so as to enable processing of the second fractions as illustrated in Fig. 6. These operations are continued until all of the fractions have been processed.
Fig. 7 illustrates the equilibration of the collecting col-umns 7.1 through 7.6 of collecting column battery 7. In this program sequence step, the collecting columns 7.1 through 7.6 are washed with water, thereby being prepared for the next cycle. 'This is done in a sequential fashion via pump 2.2, 6-way 2-position valves 3.1, 3.5, 3.6, 3.7, and 7-way 6-position valves 4.3/4.4 of collecting column battery 7. Equilibration of the collecting column batteries 8 and 9 is performed in an analogous manner. The 6-way 2-position valves 3.5 and 3.6 are switched in position, and the collecting columns 8.1 through 8.6 are equilibrated via pump 2.2, 6-way 2-position valves 3.1, 3.5, 3.6, 3.7, and 7-way 6-position valves 4.5/4.6 of collecting column bat-tery 8. Subsequently, the 6-way 2-position valves 3.6 and 3.7 are switched in position, and the collecting columns 9.1 through 9.6 are equilibrated via pump 2.2, 6-way 2-po-sition valves 3.1, 3.5, 3.6, 3.7, and 7-way 6-position valves 4.7/4.8 of co~.lecting column battery 9. Following this program sequence, the 7-way 6-position valves 4.1/4.2 of feed battery 6 are switched over to the next feed col-umns (e. g. 6.2), and t=lze entire program cycle is restarted (Sequence step 1: Equilibration of separation column 10 and venting of feed column 6.2, illustrated in Fig. 1, etc.).
After processing the second sample, the next feed column 6.3 can be switched 7_I1 the eluent stream. Because sample feed columns having undergone processing can be replaced by new ones anytime, continuous operation with an unlimited number of samples is possible.
During the first and second separation steps, chroma-tograms, retention data and spectra are collected via de-tectors 14.1, 14.2 and 14.3, processed directly in a. com-puter, and compared with data of known substances. Thus, known substances can be identified and sorted out already in on-line mode. In doubtful cases, additional data ob-tained off-line following separation and isolation can be used for identification.
Reference list 1 Low-pressure valve unit 1.1 Low-pressure valve 1.2 Low-pressure valve 1.3 Low-pressure valve 2 Pump unit 2.1 Pump 2.2 Pump 2.3 Pump 3 6-way 2-position valve 3.1 6-way 2-positi.anvalve 3.3 6-way 2-positian valve 3.4 6-way 2-position valve 3.5 6-way 2-position valve 3.6 6-way 2-position valve 3.7 6-way 2-position valve 4 7-way 6-position valve 4.1 7-way 6-position valve 4.2 7-way 6-position valve 4.3 7-way 6-position valve 4.4 7-way 6-position valve 4.5 7-way 6-position valve 4.6 7-way 6-position valve 4.7 7-way 6-position valve 4.8 7-way 6-position valve 3-way 2-position valve 5.1 3-way 2-position valve 5.2 3-way 2-position valve 5.3 3-way 2-position valve 5.4 3-way 2-position valve 5.5 3-way 2-position valve 5.6 3-way 2-position valve 5.7 3-way 2-position valve 6 Feed column battery 5.1 Feed column 5.2 Feed column 5.3 Feed column 5.4 Feed column 5.5 Feed column 5.6 Feed column 7 Collecting column battery 7.1 Collecting column 7.2 Collecting column 7.3 Collecting column 7.4 Collecting column 7.5 Collecting column 7.6 Collecting column 8 Collecting column battery 8.1 Collecting column 8.2 Collecting column 8.3 Collecting column 8.4 Collecting column 8.5 Collecting column 8.6 Collecting column 9 Collecting column battery 9.1 Collecting column 9.1 Collecting column 9.2 Collecting column 9.3 Collecting column 9.4 Collecting column 9.5 Collecting column 9.6 Collecting column Separation column il Separation columm battery 11.1 Separation column 11.2 Separation column 12 Separation column battery 12.1 Separation column 12.2 Separation column 13 Separation column battery 13.1 Separation column 13.2 Separation column 14 Detectors 14.1 Detector 14.2 Detector 14.3 Detector 15 Fraction colle,r.tor 15.1 Fraction collector 15.2 Fraction collector 15.3 Fraction collector 16 Waste 16.1 Waste 16.2 Waste 17 T-piece
A pump unit. 2 comprised of three pumps 2.1 through 2.3 is connected via 6-way 2-position valves 3.1 and 3.3 and 3-way 2-position 'valve 5.7 l..a a feed column battery 6, a separa-tion column 10 for the first separation stage, and a second separation stage comprised of three separation lines oper-able in parallel, each of which having an upstream 6-way 2-position valve :3.5, 3.6 and 3.7, respectively. In this way, it is possible to convey the mobile phase in any desired composition to any region of the device in a consecutive as well as a parallel fashion.
Each separation line h.as a collecting column battery 7, 8 and 9 and a separation column battery 11, 12 and 13. Repre-sentatively, the collecting column battery 7 includes the collecting columns 7.1. through 7.6, and the separation col-umn battery 11 includes the separation columns 11.1 and 11.2. The other two illustrated separation lines are iden-tical in configuration. Other variants including more feed columns 6.1 through 6.6 in the feed column battery 6, mul-tiple separation columns 10, more than three collecting column batteries 7, 8 and 9, each having more than six col-lecting columns, and more than three separation column bat-teries il, 12 and 13 having more than six separation col-umns per battery are also possible.
The operating sequence of the process according to the in-vention will be described in an exemplary fashion below.
Samples of mixtures of substances are dissolved in a sol-vent and added with an adsorbent each time. Subsequently, the solvent is removed using a rotary evaporator so that the adsorbent covered with sample material achieves flow-ability properties. Th.e adsorbents covered with the mixture of substances are fil7_ed into the feed columns 6.1 through 6.6 of feed column battery 6 and secured in feed column battery 6. The subsequent program sequence steps are con-trolled by a software.
The separation column 10 is equilibrated according to Fig.
1. In a parallel operation, the air is removed from the feed column battery 6. Via pump 2.3, 3-way 2-position valve 5.7 and 6-way 2-position valves 3.1 and 3.3, the air is re-moved with water from one of the dry-filled feed columns 6.1 through 6.6 to be injected next. At the same time, the separation column 10 is equilibrated with a suitable mo-bile solvent via pump 2.1 and 6-way 2-position valves 3.1 and 3.3.
Fig. 2 illustrates the separation of the mixture of sub-stances in the first separation stage on separation column and the subsequent adsorption of the fractions in a separation line inc:Luding the collecting columns 7.1 through 7.6 of collecting column battery 7.
Once the air has been removed from one of the feed columns 6.1 through 6.6, the separation program is started. Ini-tially, the 6-way ~;-position valves 3.3 and 3.5 are switched in position. Via a low-pressure valve unit 1 in-cluding the low-pressure valves 1.1 through 1.3, the compo-nents of the mobile ~ohase can be fed into the system by means of pump unit 2. The mobile phase is conveyed via the low-pressure valve 1..1 of pump 2.1 and via pump 2.1, and this system can be run both in an isocratic fashion and with a gradient. Via 6-way 2-position valve 3.3 and 7-way 6-position valves 4.1; ~4.2, the mobile phase is conveyed by pump 2.1 to that particular feed column 6.1 through 6.6 from which sample material is to be processed. The sample to be separated is transferred from one of the feed columns _ 7 -6.1 through 6.6 to the separation column 10. Via a 6-way 2-position valve 3.4 and. detector 14.1, the separated sample components discharging from separation column 10 arrive at T-piece 17, where water is mixed into the mobile phase via pump 2.2 and 6-way 2-position valve 3.1. The amount of ad-mixed water depends on the polarity of the substances to be separated. Now, as the polarity of the mobile is increased by the water, adsorption on collecting columns 7.1 through 7.6 of collecting column battery 7 is possible. Initially, adsorption is effected on the collecting column battery 7 via 6-way 2-position valve 3.5, the collecting columns 7.1 through 7.6 being loadE=_d with fractions one by one.
Fig. 3 illustrates the adsorption of additional fractions on the collecting co7_umns 8.1 through 8.6 of collecting column battery 8. Once all of the collecting columns of collecting column battery 7 are loaded with fractions, the 6-way 2-position valves 3.5 and 3.6 switch the collecting column battery 8 into the eluent stream. Now, the collect-ing columns 8.1 through 8.6 are loaded with fractions one by one.
Fig. 4 illustrates the adsorption of fractions on the col-lecting columns 9.1 through 9.6 of collecting column bat-tery 9. Once all of the collecting columns 8.1 through 8.6 of collecting column ibattery 8 are loaded with fractions, the 6-way 2-position valves 3.6 and 3.7 switch the collect-ing column battery 9 ~_nto the eluent stream. Now, the col-lecting columns 9.1 through 9.6 are loaded with fractions one by one. In the next. operating sequence step, the frac-tions adsorbed on the three collecting column batteries 7, 8 and 9 are eluted in parallel and separated further on the appropriately assigned separation column batteries 11, 12 and 13.
The separation column batteries 11, 12 and 13 are equili-brated prior to each separation. Fig. 5 illustrates the equilibration of separation column batteries 11, 12 and 13.
For equilibration, the mobile phase is conveyed to separa-tion columns 11.1 and 11.2 of separation column battery 11 via pump 2.1 and 6-waxy 2-position valves 3.1 and 3.5, re-spectively. From there, the mobile phase is conveyed to the waste via 6-way 2-posit: ion valve 3.4, detector 14.1, and a fraction collector 15.1. In a parallel operation, the sepa-ration columns 12.1 and 12.2 of the separation column bat-tery are equilibrated via pump 2.2, the 6-way 2-position valves 3.1 and 3.6, a detector 14.2, and fraction collector 15.2. Likewise, the separation columns 13.1 and 13.2 are equilibrated via pump 2.3, the 6-way 2-position valve 3.7 and 3-way 2-position valve 5.7, a detector 14.3, and a fraction collector 15.3 i.n a parallel operation.
Fig. 6 illustrates the parallel separation of fractions ad-sorbed on collecting column batteries 7, 8 and 9, using the separation column batteries 11, 12 and 13. To initiate the separation step, the mobile phase is conveyed to the col-lecting column battery 7 via pump 2.1 of pump unit 2 and 6-way 2-position valves 3.1 and 3.5. The first eluted frac-tion from collecting column battery 7 (e. g. from collecting column 7.1) is passed to separation column battery 11 via 6-way 2-position valve 3.5. There, one of the separation columns 11.1 or 11.2 c:an be switched in, optionally in a software-controlled f<~shion. Subsequently, the separated components are passed to the detector 14.1 via 6-way 2-po-sition valves 3.5 and 3.4. The software in the electronic control unit assesses the signals by peak detection, di-recting the separated components into the appropriate vials of fraction collector 15.1. Simultaneously, time control of the fraction collector 15.1 is also possible. Such time control can be activated automatically in case no peak passes the detector.
- g _ In a parallel operation, the mobile phase is conveyed to the collecting columr.~ battery 8 via pump 2.2 and 6-way 2-position valves 3.1 and 3.6. The first eluted fraction from collecting column. battery 8 (e. g. from collecting col-umn 8.1) is passed to separation column battery 12 via 6-way 2-position valve 3.6. There, one of the separation col-umns 12.1 or 12.2 can be switched in, optionally in a soft-ware-controlled fashion. The separated components are passed to the detector 14.2. In this case as well, the software assesses the signals by peak detection, subse-quently directing the separated components into the appro-priate vials of fraction collector 15.2. Similarly, the fraction collector 15.2 can be operated in a time-con-trolled fashion. Such time control can be activated auto-matically in case no peak passes the detector.
In parallel to the operations in two separation lines, the third separation line is activated with respect to initiat-ing the separation step. To this end, the mobile phase is passed to collecting column battery 9 via pump 2.3 and 3-way 2-position valve 5.7 and 6-way 2-position valve 3.7.
The first eluted fraci=ion from collecting column battery 9 (e. g. from collecting column 9.1) is passed to separation column battery 13 via valve 3.7. There, one of the separa-tion columns 13.1 or 1.3.2 can be switched in, optionally in a software-controlled fashion. The separated components are passed to the detector 14.3. The downstream fraction col-lector 15.3 is controlled as described above. After each one of the first fractions has been processed in parallel, the separation columzi batteries 11, 12 and 13 are re-equilibrated (cf . , FicT,. 5) for preparation and in order to separate the next fractions. Subsequently, the 7-way 6-po-sition valves 4.3/4.4, 4.5/4.6 and 4.7/4.8 on the collect-ing column batteries 7, 8 and 9 are switched over, so as to enable processing of the second fractions as illustrated in Fig. 6. These operations are continued until all of the fractions have been processed.
Fig. 7 illustrates the equilibration of the collecting col-umns 7.1 through 7.6 of collecting column battery 7. In this program sequence step, the collecting columns 7.1 through 7.6 are washed with water, thereby being prepared for the next cycle. 'This is done in a sequential fashion via pump 2.2, 6-way 2-position valves 3.1, 3.5, 3.6, 3.7, and 7-way 6-position valves 4.3/4.4 of collecting column battery 7. Equilibration of the collecting column batteries 8 and 9 is performed in an analogous manner. The 6-way 2-position valves 3.5 and 3.6 are switched in position, and the collecting columns 8.1 through 8.6 are equilibrated via pump 2.2, 6-way 2-position valves 3.1, 3.5, 3.6, 3.7, and 7-way 6-position valves 4.5/4.6 of collecting column bat-tery 8. Subsequently, the 6-way 2-position valves 3.6 and 3.7 are switched in position, and the collecting columns 9.1 through 9.6 are equilibrated via pump 2.2, 6-way 2-po-sition valves 3.1, 3.5, 3.6, 3.7, and 7-way 6-position valves 4.7/4.8 of co~.lecting column battery 9. Following this program sequence, the 7-way 6-position valves 4.1/4.2 of feed battery 6 are switched over to the next feed col-umns (e. g. 6.2), and t=lze entire program cycle is restarted (Sequence step 1: Equilibration of separation column 10 and venting of feed column 6.2, illustrated in Fig. 1, etc.).
After processing the second sample, the next feed column 6.3 can be switched 7_I1 the eluent stream. Because sample feed columns having undergone processing can be replaced by new ones anytime, continuous operation with an unlimited number of samples is possible.
During the first and second separation steps, chroma-tograms, retention data and spectra are collected via de-tectors 14.1, 14.2 and 14.3, processed directly in a. com-puter, and compared with data of known substances. Thus, known substances can be identified and sorted out already in on-line mode. In doubtful cases, additional data ob-tained off-line following separation and isolation can be used for identification.
Reference list 1 Low-pressure valve unit 1.1 Low-pressure valve 1.2 Low-pressure valve 1.3 Low-pressure valve 2 Pump unit 2.1 Pump 2.2 Pump 2.3 Pump 3 6-way 2-position valve 3.1 6-way 2-positi.anvalve 3.3 6-way 2-positian valve 3.4 6-way 2-position valve 3.5 6-way 2-position valve 3.6 6-way 2-position valve 3.7 6-way 2-position valve 4 7-way 6-position valve 4.1 7-way 6-position valve 4.2 7-way 6-position valve 4.3 7-way 6-position valve 4.4 7-way 6-position valve 4.5 7-way 6-position valve 4.6 7-way 6-position valve 4.7 7-way 6-position valve 4.8 7-way 6-position valve 3-way 2-position valve 5.1 3-way 2-position valve 5.2 3-way 2-position valve 5.3 3-way 2-position valve 5.4 3-way 2-position valve 5.5 3-way 2-position valve 5.6 3-way 2-position valve 5.7 3-way 2-position valve 6 Feed column battery 5.1 Feed column 5.2 Feed column 5.3 Feed column 5.4 Feed column 5.5 Feed column 5.6 Feed column 7 Collecting column battery 7.1 Collecting column 7.2 Collecting column 7.3 Collecting column 7.4 Collecting column 7.5 Collecting column 7.6 Collecting column 8 Collecting column battery 8.1 Collecting column 8.2 Collecting column 8.3 Collecting column 8.4 Collecting column 8.5 Collecting column 8.6 Collecting column 9 Collecting column battery 9.1 Collecting column 9.1 Collecting column 9.2 Collecting column 9.3 Collecting column 9.4 Collecting column 9.5 Collecting column 9.6 Collecting column Separation column il Separation columm battery 11.1 Separation column 11.2 Separation column 12 Separation column battery 12.1 Separation column 12.2 Separation column 13 Separation column battery 13.1 Separation column 13.2 Separation column 14 Detectors 14.1 Detector 14.2 Detector 14.3 Detector 15 Fraction colle,r.tor 15.1 Fraction collector 15.2 Fraction collector 15.3 Fraction collector 16 Waste 16.1 Waste 16.2 Waste 17 T-piece
Claims (7)
1. A process for the rapid liquid-chromatographic separa-tion and identification of substances, characterized in that mixtures of substances are preseparated in the first stage of a software-controlled rapid liquid-chro-matographic two-stage separation, the preseparated fractions deposited in collecting columns are subjected to a parallel final separation in at least two separa-tion lines in the second stage, and the finely sepa-rated fractions are identified and isolated in parallel operations each time.
2. The process according to claim 1, characterized in that the preseparation of mixtures of substances is per-formed consecutively in the first separation stage, and fine separation in the second stage is performed con-secutively and/or in parallel.
3. The process according to claim 1 or 2, characterized in that at least one detector (14.1) is used both after the first separation stage and after the second separa-tion stage.
4. The process according to any of claims 1 to 3, charac-terized in that the substances separated and isolated in the separation lines are subjected to an additional purification procedure, particularly an adsorptive pu-rification.
5. A device for the rapid liquid-chromatographic separa-tion and identification of substances, said device con-sisting of a plurality of separation columns and col-lecting columns, as well as feed systems, detectors, and fraction collectors, the interaction of which can be controlled through a central control unit, charac-terized in that a plurality of parallel liquid-chroma-tographic separation lines, each one consisting of a combination of separation column batteries (11, 12, 13) and collecting column batteries (7, 8, 9), detector units (14), and fraction collector units (15), are ar-ranged downstream o.f at least one separation column (10), that a pump unit (2) consisting of three pumps (2.1, 2.2, 2.3) to convey the mobile phase is function-ally connected both with the separation column (10) and the separation lines, and that software-switchable multi-way valves are arranged between the individual functional units.
6. The device according to claim 5, characterized in that each separation line has a multi-way valve (3.5, 3.6, 3.7) arranged upstream thereof.
7. The device according to claim 5 or 6, characterized in that additional collecting columns are arranged down-stream of the separation lines.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE19847439.3 | 1998-10-08 | ||
DE1998147439 DE19847439C2 (en) | 1998-10-08 | 1998-10-08 | Method and device for liquid chromatographic separation of substance mixtures and identification of substances |
PCT/EP1999/007542 WO2000022429A1 (en) | 1998-10-08 | 1999-10-08 | Method and device for the rapid liquid chromatographic separation of substance mixtures and for the identification of substances |
Publications (1)
Publication Number | Publication Date |
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CA2346358A1 true CA2346358A1 (en) | 2000-04-20 |
Family
ID=7884498
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA002346358A Abandoned CA2346358A1 (en) | 1998-10-08 | 1999-10-08 | Method and device for the rapid liquid chromatographic separation of substance mixtures and for the identification of substances |
Country Status (6)
Country | Link |
---|---|
EP (1) | EP1119767A1 (en) |
JP (1) | JP2002527748A (en) |
AU (1) | AU6469599A (en) |
CA (1) | CA2346358A1 (en) |
DE (1) | DE19847439C2 (en) |
WO (1) | WO2000022429A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102803948A (en) * | 2009-06-09 | 2012-11-28 | 通用电气健康护理生物科学股份公司 | Automated fluid handling system |
Families Citing this family (7)
Publication number | Priority date | Publication date | Assignee | Title |
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WO2001068216A2 (en) * | 2000-03-13 | 2001-09-20 | Eli Lilly And Company | Parallel preparative automated isolation system |
JP4615101B2 (en) * | 2000-08-08 | 2011-01-19 | 昭光サイエンティフィック株式会社 | Purification preparative equipment |
DE20020555U1 (en) | 2000-12-05 | 2001-05-17 | LC Tech GmbH, 84405 Dorfen | Device for generating a predetermined volume of pre-cleaned components of solution mixtures |
JP2002350412A (en) * | 2001-05-23 | 2002-12-04 | Nobuo Tanaka | Multidimensional high performance liquid chromatograph |
CA2536809A1 (en) * | 2003-09-05 | 2005-03-17 | Sumitomo Chemical Company, Limited | Liquid chromatographic apparatus |
US9608929B2 (en) | 2005-03-22 | 2017-03-28 | Live Nation Entertainment, Inc. | System and method for dynamic queue management using queue protocols |
CN102441294A (en) * | 2010-09-30 | 2012-05-09 | 中国科学院昆明植物研究所 | Separation and preparation chromatograph containing series static axial compression (SAC) preparative chromatographic columns |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
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US5198115A (en) * | 1991-07-09 | 1993-03-30 | Analytical Bio-Chemistry Laboratories, Inc. | Integrated instrument for supercritical fluid sample extraction, sample separation and concentration |
US5670054A (en) * | 1996-04-04 | 1997-09-23 | Warner Lambert Company | Method and system for identification, purification, and quantitation of reaction components |
DE19641210A1 (en) * | 1996-09-25 | 1998-04-02 | Analyticon Ag Biotechnologie P | Separation of complex mixtures, e.g. of natural products |
EP0946236B1 (en) * | 1996-09-25 | 2002-05-15 | SEPIAtec GmbH | Hplc-based device and method for separating high complex substance mixtures |
-
1998
- 1998-10-08 DE DE1998147439 patent/DE19847439C2/en not_active Expired - Fee Related
-
1999
- 1999-10-08 AU AU64695/99A patent/AU6469599A/en not_active Abandoned
- 1999-10-08 EP EP99952538A patent/EP1119767A1/en not_active Withdrawn
- 1999-10-08 WO PCT/EP1999/007542 patent/WO2000022429A1/en not_active Application Discontinuation
- 1999-10-08 JP JP2000576276A patent/JP2002527748A/en not_active Withdrawn
- 1999-10-08 CA CA002346358A patent/CA2346358A1/en not_active Abandoned
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102803948A (en) * | 2009-06-09 | 2012-11-28 | 通用电气健康护理生物科学股份公司 | Automated fluid handling system |
CN102803948B (en) * | 2009-06-09 | 2015-11-25 | 通用电气健康护理生物科学股份公司 | Robotization fluid handling system |
Also Published As
Publication number | Publication date |
---|---|
WO2000022429A1 (en) | 2000-04-20 |
DE19847439C2 (en) | 2001-10-18 |
EP1119767A1 (en) | 2001-08-01 |
AU6469599A (en) | 2000-05-01 |
JP2002527748A (en) | 2002-08-27 |
DE19847439A1 (en) | 2000-04-20 |
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