WO1999017104A1 - Dispositif et procede servant a mesurer la radioactivite en chromatographie liquide - Google Patents

Dispositif et procede servant a mesurer la radioactivite en chromatographie liquide Download PDF

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Publication number
WO1999017104A1
WO1999017104A1 PCT/US1998/020324 US9820324W WO9917104A1 WO 1999017104 A1 WO1999017104 A1 WO 1999017104A1 US 9820324 W US9820324 W US 9820324W WO 9917104 A1 WO9917104 A1 WO 9917104A1
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Prior art keywords
radioactivity
eluate
flow
detector
fraction
Prior art date
Application number
PCT/US1998/020324
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English (en)
Inventor
Dian Y. Lee
Original Assignee
Lee Dian Y
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Lee Dian Y filed Critical Lee Dian Y
Priority to AU13596/99A priority Critical patent/AU1359699A/en
Publication of WO1999017104A1 publication Critical patent/WO1999017104A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • 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
    • G01N30/62—Detectors specially adapted therefor
    • G01N2030/77—Detectors specially adapted therefor detecting radioactive properties
    • 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
    • G01N30/84—Preparation of the fraction to be distributed
    • G01N2030/8411—Intermediate storage of effluent, including condensation on surface
    • 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
    • G01N30/80—Fraction collectors
    • G01N30/82—Automatic means therefor

Definitions

  • This invention generally relates to an apparatus and method for measurement of radioactivity in liquid chromatography.
  • the present invention provides a stop-flow radioactivity measurement apparatus and method designed to satisfy the aforementioned needs.
  • the stop-flow radioactivity measurement is carried out by a stop-flow system that stops and resumes the flow of eluate repeatedly in order to measure the radioactivity in each desired fraction or portion of an eluate in a stationary manner. Since the measurement time devoted for any fractions, peaks, or regions of a chromatogram is controllable, the accuracy and sensitivity of the measurement will increase dramatically. Furthermore, no regions in the chromatogram will be missed for accurate stationary measurement, thus providing a true on-line radioactivity detection method with high accuracy and sensitivity for liquid chromatography. With this invention, the fraction collection/LSC method used in the prior art can be completely eliminated, thus reducing the radioactive wastes and improving the working environments and productivity. Also, the cost associated with the implementation of current invention is much less than that of the prior art procedure.
  • the present invention relates to apparatus and method for measurement of radioactivity in liquid chromatography, which include the operative steps of (a) stopping the flow of the eluate and (b) measuring the radioactivity of the eluate present in the radioactivity detector in a stationary manner while the flow of the eluate stops, whereby the radioactivity in the eluate will be measurable for a longer period of time.
  • the invention may also include the step of resuming the flow of the eluate from liquid chromatography to introduce the following fraction into the radioactivity detector for stationary measurement.
  • the stop-flow operation can be activated by a timed sequence, radioactivity level detected by the radioactivity detector or an external signal.
  • the flow cell can be cleaned, if necessary, by flushing the cell with a solvent or liquid scintillation cocktail prior to the introduction of the following fraction.
  • the current invention also provides a method which includes the operative steps of (a) collecting fractions of an eluate from liquid chromatography in a storage device and (b) feeding the fractions individually back into the radioactivity detector for counting in a stationary manner.
  • the operation as to when and where to collect fractions can be activated by either a timed sequence or other signals such as radioactivity signal observed in the radioactivity detector.
  • FIG. 1 shows the first embodiment of the stop-flow radioactivity counting apparatus pursuant to the present invention
  • FIG. 2 shows the second embodiment of the stop-flow radioactivity counting apparatus of the present invention
  • FIG. 3 shows the detailed sequences for a typical stop-flow radioactivity counting process of the present invention
  • FIG. 4 shows the detailed sequences for another mode of operation for counting only the peaks in the present invention.
  • FIG. 1 there is schematically shown the stop-flow radioactivity counting apparatus for detecting radioactivity from an eluate in liquid chromatograph.
  • the apparatus basically comprises of pump 1 for delivering mobile phase (s), column 10 for fractionation of components of a sample, radioactivity detector 17 for measurement or counting of radioactivity, and control arrangement 35 for data collection and communication with various devices involved in the present invention.
  • Pump 1 can include one or more pumps for delivering a isocratic (one composition of solvent throughout the entire analysis or run) or a gradient of mobile phase.
  • pump 1 is being controlled by control arrangement 35 for the stop-flow operation.
  • Column 10 is usually a liquid chromatography column which comprises of a tubing and a packed stationary phase for fractionation of components of samples.
  • Column 10 can be a analytical, narrow- bore, micro-bore, or semi-preparative LC column (LC equals to liquid chromatography) .
  • Radioactivity detector 17 can be a radioactivity flow-through detector suitable for high pressure liquid chromatography. Radioactivity detector 17 can also be a normal liquid scintillation counter as long as the flow cell is placed between two photomultiplier tubes. Radioactivity detector 17 can have a liquid cell or a solid cell or the combination thereof. The counting data from radioactivity detector 17 can be either digital format or a analog format which will be digitized in the control arrangement.
  • Control arrangement 35 communicates with pump 1 through signal line 34, with radioactivity detector 17 through signal line 14, with sample injector 3 through signal line 4, valve 47 through signal line 6, with pump 26 through signal line 48, with detector 12 through signal line 13, valve 18 through signal line 46, and fraction collector 22 through signal line 16.
  • Valve 47 is preferred in order to stop the flow of mobile phase completely to achieve the best performance in separation under stop-flow operation.
  • Valve 47 can be a on/off valve or switching valve which enables the flow at one position and disables or block the flow of mobile phase (s) at another position, which is preferably disposed immediately after column 10.
  • valve 47 can also be disposed immediately after radioactivity detector 17 if radioactivity detector 17 and other devices between radioactivity detector 17 and column 10 can tolerate the pressure during the stop-flow operation.
  • Pump 26 is used for pumping either scintillation fluid or non- radioactive solvent (s) or a solvent mixture in container 24 through line 25 and to radioactivity detector 17 through line 27 for either liquid scintillation counting purpose or flushing out the residual radioactivity residing inside the flow cell of radioactivity detector 17. No scintillation fluid is needed for a solid cell.
  • Sample injector 3 disposed between pump 1 and column 10 via lines 50 and 37, is either a manual injector (such as a loop on a multiple port switching valve) or automatic injector (auto-sampler) .
  • Detector 12 such as a U (ultraviolet) detector, can be connected on-line between valve 47 and radioactivity detector 17 through lines 42 and 43.
  • the connection lines between the column 10 and valve 47 is 41.
  • Fraction collector 22 and waste container 20 are connected through lines 21 and 19 from valve 18 which is connected to the outlet of radioactivity detector 17 through line 28.
  • the stop-flow radioactivity counting method can be used in several different modes using the embodiment of FIG. 1.
  • the first mode is to measure radioactivity by counting every fraction eluting from an entire LC run.
  • the second mode is to count only the peaks which exceed predefined criteria (either by a threshold or other algorithms) .
  • the third mode is to count only the regions of interest of the LC run. Any combination of these three modes can be used as well. Here are shown only the details of those common modes of operation. However, many variations of this stop-flow counting method are possible without departing from the spirit of the present invention.
  • FIG. 3 shows the beginning part of the operation sequences for a typical stop-flow radioactivity counting method with a predefined time interval for fractions.
  • FIG. 3A shows the change of the mobile phase composition in pump 1 under a linear gradient. If a isocratic mobile phase is used, the line in FIG. 3A becomes a horizontal line parallel to the time (horizontal) axis.
  • FIG. 3B shows the flowrate of mobile phase delivered by pump 1.
  • FIG. 3C shows the flowrate of pump 26.
  • FIG. 3D shows the radioactivity counting operation in radioactivity detector 17.
  • FIG. 3E shows the results of the stop-flow radioactivity counting representing for the radioactivity in fractions FI through F5.
  • valve 47 Before the sample is injected, valve 47 is at the open position and the pump 1 is at the initial conditions of the liquid chromatography.
  • the gradient is divided into equal or different time intervals or fractions for counting radioactivity by radioactivity detector 17 in a stationary manner.
  • the first five fractions (FI through F5) are shown in FIG. 3B.
  • Fraction FI contains the eluate collected in the flow cell (either a solid or a liquid cell) of radioactivity detector 17 between time points TO and Tl .
  • Fraction F2 is the eluate collected from time points Tl through T2, etc.
  • a signal is generated and sent through signal line 4 to control arrangement 35 which triggered pump 1.
  • the injection signal can be sent directly from sample injector 3 to pump 1 through signal line 49 for triggering of the run.
  • pump 1 is triggered, the solvent elution or gradient will start.
  • the sample is pushed onto column 10 for fractionation and the eluate is collected in the flow cell (not shown) of radioactivity detector 17. If a liquid cell is used, pump 26 also starts pumping liquid scintillation fluid which is mixed with eluate from line 43 before the liquid cell. If a solid cell is used, pump 26 does not pump anything into radioactivity detector 17.
  • control arrangement 35 When time point Tl is encountered, control arrangement 35 will send a stop-flow signal to pump 1 to stop the flow of the mobile phase through signal line 34, and both flow and gradient are stopped or paused. At the same time, if valve 47 is used, control arrangement 35 also sends a signal through line 6 to turn valve 47 to the close or off position in order to stop the flow of mobile phase completely.
  • radioactivity detector 17 starts measuring (or counting) the radioactivity contained in the flow cell (from time points Tl to a) ) .
  • the radioactivity is measured by detecting the flash of photons generated by the interaction of radiation particles (such as beta particles from carbon-14 isotopes, etc.) with either liquid or solid scintillator by using usually a pair of photomultiplier tubes inside radioactivity detector 17.
  • This process is also called counting radioactivity because the measurement process is actually counting the radiation events associated with the decay of radioactive isotopes. Since the counting time can be controlled by the user, any desired levels of accuracy on the radioactivity counting or measurement can be achieved. The more counts are accumulated or the longer the fraction is counted, the more accurate results one will obtain from radioactivity detector 17 for the fraction. This is due to the nature of the radiation which is a random process and follows the distribution curve of the Poison distribution statistically. The counts accumulated from this entire counting period is calculated as the mean to represent the radioactivity of fraction FI . The percent error of the counting, together with other parameters, can be calculated as well to indicate the performance of the counting process .
  • time point a After accumulating the counts for either a predefined period of time (e.g. 2 min) or based on predefined criteria for counting accuracy or counting errors, time point a) is encountered.
  • pump 26 In order to remove the residual radioactivity residing inside the flow cell and eliminate any possible cross contamination for the next incoming fraction, pump 26 starts pumping at the same or different flowrate. Pump 26 will pump liquid scintillation fluid if a liquid cell is used. If a solid cell is used, pump 26 will pump a non-radioactive solvent or solvent mixture.
  • the pumping operation of pump 26 between points a) and b) can be eliminated if the volume of the fraction (for a solid cell) or the combined volume of the fraction and the liquid scintillation fluid (for a liquid cell) approximately equals to the cell volume of radioactivity detector 17.
  • This pumping operation is designed to remove any residual radioactivity out of the either solid or liquid cells before the introduction of the next new fraction (e.g. fraction F2) .
  • the flow and gradient (if any) , of pump 1 is resumed for another cycle. If a solid cell is used, pump 26 stops pumping any solvent.
  • this stop-flow radioactivity counting method cycle includes the following steps: collecting the eluate into the flow cell; stopping the flow and stopping (pausing) the gradient (if any) ; counting the radioactivity inside the flow cell in a stationary manner; removing the residual radioactivity from the flow cell; and finally resuming the flow and gradient (if any) of pump 1 and pump 26 (if a liquid cell is used) .
  • the gradient is resumed, it starts from the point where it was stopped previously. This will ensures the best performance in separation and resolution comparable to the prior art continuous run or analysis.
  • next cycle continues for next fraction F2, etc.
  • FIG. 3 we observed five counting cycles (from time points TO through n) ) , each of which counts the corresponding fraction (FI through F5) .
  • the results of the counting cycles are shown in the radio-chromatogram in FIG. 3E, which has five data points, each of which representing the counting results from corresponding fractions (FI through F5) .
  • a peak is accurately detected in fraction F4 at time point T4 of FIG. 3E .
  • the time scale on FIG. 3A is the time scale for a continuous run.
  • the time scales in FIG. 3B through FIG. 3D are the actual time scales during the stop-flow counting process.
  • 3E is the reconstructed time scale by eliminating the time spent during the stop-flow counting cycles.
  • the reconstructed radio-chromatogram represents the radioactivity distribution of fractions in FIG. 3A.
  • FIG. 4 shows the sequences for counting only the peaks which exceed the predefined criteria (either by a threshold or other algorithms) .
  • the threshold is used to detect the peaks.
  • FIG. 4A shows the radio-chromatogram of a sample containing two radioactive peaks if the sample were run under normal method in the prior art (i.e. continuous run with a on- line radioactivity flow-through detector) .
  • the threshold is preset to recognize any peaks for stop-flow counting.
  • FIG. 4B shows the flowrate of pump 1.
  • FIG. 4C shows the flowrate of pump 26 for delivering liquid scintillation fluid. If a solid cell is used, pump 26 only delivers the non-radioactive flushing solvent (s) after counting of each fraction and before introduction of next new fraction.
  • FIG. 4D shows the counting operation in radioactivity detector 17. Basically, all the data from the entire run are being used to generate the radio-chromatogram as shown in FIG. 4E .
  • the data acquired while the pump 1 stops is calculated as a single data point (as the mean) to represent the corresponding fraction inside the flow cell.
  • FIG. 4E shows the reconstructed radio-chromatogram obtained from this counting method.
  • a signal is generated and sent to control arrangement 35 which in turn sends a signal to pump 1 to start the flow and gradient (if . any) .
  • pump 26 starts pumping liquid scintillation fluid to radioactivity detector 17 if a liquid cell is used. If a solid cell is used, pump 26 does not pump anything.
  • radioactivity detector 17 detects Peak 1 (based on either a higher radioactivity than the preset threshold or other algorithms such as changes in slope, etc.), a fraction of eluate, the size of which is determined either by the predefined time intervals or levels of radioactivity detected or detection of the end of the peak.
  • the control arrangement will stop the flow of pump 1 and the gradient (if any) and count the radioactivity inside the flow cell for a predefined period of time or to accumulate enough counts to reach certain levels of counting accuracy.
  • fraction F6, containing eluate from time points TO' through Tl' is counted first (from time points Tl' to p) ) .
  • the accumulated counting data are calculated as the mean which represents the radioactivity level of fraction F6, which is shown at time point x) in FIG. 4E .
  • the content of the flow cell is flushed out by solvent (s) delivered by pump 26 between time points p) and q) .
  • solvent (s) delivered by pump 26 between time points p) and q) .
  • the flow and gradient (if any) of pump 1 is resumed until time point r) , where fraction F7 (eluate from time points Tl' to T2' ) is ready for counting.
  • Fraction F7 is subjected to a similar counting cycle as shown for fraction F6 from time points r) to t) , resulting the data at time point y) calculated as the mean.
  • the flow of pump 1 continues until the next peak (Peak 2) is encountered.
  • Peak 2 is narrow enough so that only one fraction (fraction F8) containing eluate from time points T3' through T4' is needed to count the entire peak.
  • the entire Peak 2 is counted in a stationary manner in the counting and flushing cycle between time points of u) and w) .
  • the obtained data is presented at time point z) in the radio-chromatogram of FIG. 4E .
  • the accumulated counts for each fraction is calculated as the mean to represent the radioactivity of that fraction and positioned in the radio-chromatogram at the center position of that fraction on the time axis.
  • radioactivity data from fraction F6 is positioned at the center (time point x) ) in the fraction between time points TO' and Tl' on the time axis and data from fraction F7 is positioned at the center between time points T3' and T4', etc.
  • the counting data which are obtained by radioactivity detector 17 during the regions where no peaks are detected and counted, will be treated as raw data in the normal manner as used in the prior art.
  • This counting cycle can be applied to all the peaks detected in a LC run.
  • the accurate quantitation of radioactive peaks are thus possible by my stop-flow counting method.
  • a broad peak can be counted in several consecutive fractions.
  • a sharp or narrow peak can be counted in a single fraction. Since the counting time can be controlled, any desired levels of counting accuracy and sensitivity can be obtained. Furthermore, since the fraction size is controllable, the desired resolution of radio-chromatogram can be achieved for any peaks .
  • a region or regions of the interest during a LC chromatogram can be measured accurately for radioactivity.
  • the control arrangement will send signals to stop the flow and pause the gradient through signal lines 34 and 6 and radioactivity detector 17 starts the counting process in a stationary manner for a period of time.
  • each region can be counted as more than one fraction.
  • fraction collector 22 and waste container 20 can be used to collect useful radioactive components detected by radioactivity detector 17.
  • Valve 18 is a three- way valve which is controlled by control arrangement 35 through signal line 46.
  • Fraction collector 22 is controlled also by control arrangement 35 through signal line 16.
  • the components of interest in the eluate can be collected using fraction collector 22.
  • the fraction collection can have several modes of operation. The first one is to collect peak(s). When a peak is detected in radioactivity detector 17 based on predefined criteria, the eluate is directed to the fraction collector through the valve after a delay time which compensates the time needed for eluate traveling from the outlet of radioactivity detector 17 to valve 18. When the end of the peak is detected, the eluate is directed to the waste container through valve 18 after the delay time.
  • the second mode is collection of predefined regions of interest. Again, when the beginning of region (s) of interest is encountered, the control arrangement will send signals through line 46 which turns the valve toward the fraction collector.
  • valve When the end of the regions of interest is encountered, the valve is turned back to the waste container by control arrangement 35 through signal line 46 for collection of waste.
  • This is an excellent tool to collect the needed peaks or components for further analysis by other means such as mass spectrometry or NMR (nuclear magnetic resonance) .
  • This method is also good for waste management where separation of high level of radioactive wastes from low level or non-radioactive wastes are desired, thus saving cost in waste disposal.
  • Coupling of line 21 in FIG. 2 to a LC/MS (liquid chromatography-mass spectrometry) is also possible and will provide a unique way of analyzing peaks more efficiently.
  • This method is especially useful for a LC/MS/MS experiment where several modes of operation are needed for a single peak which is usually eluting from a column too fast and several runs of the same sample are needed to obtain the desired results.
  • stop-flow counting method the peak(s) can be stopped allowing MS to have enough time to conduct a series of experiments on a single peak detected by the method of present invention.
  • FIG. 2 shows another embodiment of the present invention.
  • valves 9, 2 , 44 and storage devices 31 and 33 are incorporated.
  • Valves 2 and 44 are two position switching valves (such as the four-port switching valves from Valco Instruments Co. Inc., P.O. Box 55603, Houston, TX 77255) .
  • Valve 9 is a multiple position valve (such as the one similar to Valco stream selection valve for selection of up to six LC columns, part number: DCST6UW) for column selection.
  • Storage devices 31 and 33 are multiple position valves (such as the one similar to the multiposition stream selection valve from Valco Instruments Co. Inc., part number: DCST16MWEY) .
  • Storage device 31 or 33 comprises of multiple capillary tubing for storage of eluate fractions. Depending on the desired number of capillary tubing used, one such device can be sufficient. If more than one storage devices are used, they can be connected in series or parallel to each other.
  • Valve 2 is disposed through line 37 immediately before valve 9 where multiple LC columns can be connected.
  • column 10 is connected through lines 41 and 40 and column 11 is connected through lines 38 and 39.
  • Valve 44 is disposed between radioactivity detector 17 through line 28 and valve 18 through line 45. Valves 2 and 44 is connected through line 8.
  • Valve 2 is connected with control arrangement 35 through signal line 5.
  • Storage devices 31 and 33 are connected in series and connected to valve 2 and 44 through lines 23, 32 and 7.
  • the stop-flow radioactivity counting method using embodiment of FIG. 2 is shown as follows. Since storage devices 31 and 33 can store the fractions of the eluate into the capillary tubing, the eluate can be collected and stored in the storage devices and counted after the LC run is finished. The size and length of the capillary tubing on storage devices 31 and 33 can vary depending on the fraction size and total number of fractions per LC run.
  • valve 2 is at the position connecting lines 36 and 37 and valve 44 connecting lines 28 and 23.
  • the eluate flows through the following lines and devices sequentially: 1, 50, 3, 36, 2, 37, 9, 40, 10, 41, 42, 12 (if any), 43, 17, 28, 23, 33, 32, 31, 7, 8, 45, 18, 19, and finally arrives waste container 20.
  • a sample injection signal is generated and sent through line 4 to the control arrangement.
  • Control arrangement 35 then sends a signal through line 34 to trigger pump 1 to start the run.
  • the control arrangement 35 will turn storage device 33 to the first position to collect eluate which fills the first capillary tubing through signal line 29.
  • the control arrangement 35 will turn the storage device 33 into its second position to collect the second fraction, etc.
  • the control arrangement 35 will turn storage device 31 through signal line 30 to its first position and so on until all the desired fractions are collected.
  • each of the fractions collected and stored in storage devices 33 and 31 is individually pushed back to radioactivity detector 17 for radioactivity counting in a stationary manner.
  • the control arrangement 35 will first turn valves 2, 44 (through signal line 15) and 9 such way that the flow path of the solvent from pump 1 is going through the following lines and devices: 1, 50, 3, 36, 2, 7, 31, 32, 33, 23, 8, 2, 37, 9 (without going through any columns) , 42, 12, 43, 17, 28, 44, 45, 18, 19, and waste container 20. Then the control arrangement will trigger pump 1 to deliver mobile phase at certain flowrate.
  • the control arrangement 35 will turn storage device 33 to its first position to feed the first fraction back to radioactivity detector 17.
  • a delay time is calculated by the control arrangement based on the flowrate of pump 1 and volume of the flowpath from storage device 33 through lines or devices of 23, 44, 8, 2, 37, 9, 42, 12, and 43. If a liquid cell is used, the LSC pump will start pumping the liquid scintillation fluid preferably just before the fraction arrives the flow cell. After the delay time, the flow is stopped by control arrangement 35 and the fraction is counted in radioactivity detector 17 in a stationary manner for a period of time and the data is collected through line 14.
  • the counting time is predefined or judged by the control arrangement based on, for example, predefined intervals, the counting accuracy or magnitudes of counting errors.
  • pump 26 will flush out any residual radioactivity inside the liquid cell at preferably a higher flowrate in order to save time.
  • pump 26 will be pumping a non-radioactive solvent or a solvent mixture instead of liquid scintillation fluid which is not usually comparable with a solid cell. If the volume of the flow cell equals to the volume of the fraction in case of a liquid cell or the combined volume of the fraction and the liquid scintillation fluid in case of a liquid cell, this flushing step can be eliminated since the following fraction will replace the previous fraction inside the flow cell. This is the counting cycle for counting one fraction stored in storage device 33.
  • control arrangement 35 will send a signal to pump 1 to resume the flow of the mobile phase and repeat the cycle described above to count the radioactivity of the subsequent fractions one by one. After all the fractions are counted, a radio-chromatogram containing all the counting results, each of the data points representing the counting results of each fraction, can be obtained.
  • the desired radioactive peaks or regions of interest can be collected into fraction collector 22 which is controlled by control arrangement 35 through line 16.
  • control arrangement 35 When peak(s) is detected in radioactivity detector 17, control arrangement 35 will turn valves 44 and 18 such way that the eluate will go through: 28, 44, 45, 18, 21 and finally to fraction collector 22. If more than one peaks are being collected, the control arrangement will advance the fraction collector to the next tube or vial by sending a signal through line 16. Same procedures can be used for collecting region (s) of interest. Moreover, more than one fraction can be collected from one region of interest.
  • control arrangement 35 will turn valves 44, 2, 9, storage devices 33 and 31 in such way that before the run the mobile phase will use the following flow path going sequentially through parts of 1, 50, 3, 36, 2, 37, 9, 40, 10, 41, 42, 12, 43, 17, 28, 44, 45, 18, 19, and 20.
  • control arrangement 35 When the beginning of the radioactive peaks or regions of interest is encountered, control arrangement 35 turns storage devices 33 and 31 to their first positions and will turn valve 44 to connect lines 28 and 23, the peak or the region of interest is collected into the first capillary tubing of storage device 33. If the first capillary tubing is full and the peak or region of interest is not finished, control arrangement 35 will turn storage device 33 into the next position to continue the collection, etc. When the peak or the region of interest is finished, control arrangement 35 will turn valve 44 back to connect line 28 to line 45 until the next coming peak or region of the interest, if any. This process is repeated until all the peaks or regions of interest is collected and stored in the capillary tubings on storage device 33 and/or 31.
  • the collected peaks or regions of interest on storage devices 33 or 31 can be counted for radioactivity in a stationary manner in a similar way as described above for counting fractions stored on storage devices 33 or 31.
  • the content of each capillary tubing can be counted once or divided into many portions and counted separately.
  • the flow cell can be flushed with a organic solvent or solvent mixture delivered by pump 26. This will ensure the elimination of the possible cross contamination resulting from the residual radioactivity from the previously counted sample.
  • Part Name No. Part Name No. Part Name

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Abstract

Procédé et dispositif d'arrêt d'écoulement servant à mesurer la radioactivité dans un éluat chromatographique liquide, ce qui consiste à arrêter l'écoulement et à le laisser reprendre, de façon à pouvoir mesurer des parties souhaitées de l'éluat de manière précise et fixe. On fait passer le spécimen à travers une colonne chromatographique (10) vers un détecteur de radioactivité (17) par l'intermédiaire d'un clapet (47) sous la commande d'une unité de commande (35). Ce clapet (47) est fermé en réaction à des signaux émis par une minuterie, par le détecteur de radioactivité (17) ou par un autre détecteur (12) afin d'arrêter l'écoulement de l'éluat et de permettre la mesure fixe. Dans un deuxième mode de réalisation, des fractions d'éluat peuvent être accumulées dans un accumulateur capillaire préalablement à leur passage vers le détecteur de radioactivité (17) afin d'effectuer une mesure fixe.
PCT/US1998/020324 1997-10-01 1998-09-25 Dispositif et procede servant a mesurer la radioactivite en chromatographie liquide WO1999017104A1 (fr)

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AU13596/99A AU1359699A (en) 1997-10-01 1998-09-25 Apparatus and method for radioactivity measurement in liquid chromatography

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US6063797P 1997-10-01 1997-10-01
US60/060,637 1997-10-01
US4424398A 1998-03-19 1998-03-19
US09/044,243 1998-03-19

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6546786B2 (en) 2000-07-27 2003-04-15 Dian Y. Lee Methods and apparatus for detection of radioactivity in liquid samples
EP1467204A2 (fr) * 2003-04-09 2004-10-13 ISCO, Inc. Procédé et dispositif d'amélioration du rapport signal-bruit en chromatographie
WO2023080870A1 (fr) * 2021-11-05 2023-05-11 Eczacibaşi Monrol Nükleer Ürünler Sanayi̇ Ve Ti̇caret Anoni̇m Şi̇rketi̇ Colonne de séparation à compartiment séparé

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US4704531A (en) * 1983-08-12 1987-11-03 Laboratorium Prof. Dr. Rudolph Berthold Method and apparatus for measuring the radioactivity in column eluates
US5099129A (en) * 1984-11-22 1992-03-24 Pullan Brian R Multiple sample radioactivity detector
US5283036A (en) * 1991-02-11 1994-02-01 Bruker Analytische Messtechnik Gmbh Apparatus for coupled liquid chromatography and nuclear magnetic resonance spectroscopy measurements
US5591644A (en) * 1990-05-11 1997-01-07 Albert Einstein College Of Medicine Of Yeshiva University Sensitive on line assay of chromatographic effluent

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US4704531A (en) * 1983-08-12 1987-11-03 Laboratorium Prof. Dr. Rudolph Berthold Method and apparatus for measuring the radioactivity in column eluates
US5099129A (en) * 1984-11-22 1992-03-24 Pullan Brian R Multiple sample radioactivity detector
US5591644A (en) * 1990-05-11 1997-01-07 Albert Einstein College Of Medicine Of Yeshiva University Sensitive on line assay of chromatographic effluent
US5283036A (en) * 1991-02-11 1994-02-01 Bruker Analytische Messtechnik Gmbh Apparatus for coupled liquid chromatography and nuclear magnetic resonance spectroscopy measurements

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6546786B2 (en) 2000-07-27 2003-04-15 Dian Y. Lee Methods and apparatus for detection of radioactivity in liquid samples
EP1467204A2 (fr) * 2003-04-09 2004-10-13 ISCO, Inc. Procédé et dispositif d'amélioration du rapport signal-bruit en chromatographie
EP1467204A3 (fr) * 2003-04-09 2005-02-09 ISCO, Inc. Procédé et dispositif d'amélioration du rapport signal-bruit en chromatographie
WO2023080870A1 (fr) * 2021-11-05 2023-05-11 Eczacibaşi Monrol Nükleer Ürünler Sanayi̇ Ve Ti̇caret Anoni̇m Şi̇rketi̇ Colonne de séparation à compartiment séparé

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