Apparatus for preparative supercritical fluid chromatography

[0001] This application claims the right to benefit from the application of preliminary supercritical flow chromatography according to application number 60 / 154,038, filed on September 16, 1999.

PROBLEM TO BE SOLVED: To recover a highly purified component of interest from a sample composed of a simple or complex mixture of a plurality of components, there is a basic industrial demand. Many techniques have been developed to meet this demand. The technique of choice for the non-volatile components that can be eluted has been liquid elution chromatography.

[0003] Analytical engineers have several goals in adopting preliminary elution chromatography (a method of separating a mixture into components). First, they want each ingredient of interest to be as pure as possible. Second, they want to recover the maximum amount of each feature of interest. Third, they want to process continuously, and if possible, remove samples unrelated to the first sample as quickly as possible to get one that is free of impurities. Finally, it is often desirable to have a solution that can be rapidly purified, as a solvent-free component, or in a well-structured solution that contains or contains the first collection solvent. Collecting the sample as a non-solvent.

[0004] In normal phase chromatography, only an organic solvent or a mixture thereof is used as the eluent, and a typical volume ratio of several tens of milliliters out of 100 milliliters is common. To recover the residue component of interest, the portion must then be evaporated for a considerable length of time.

[0005] In reverse phase chromatography, a mixture of an organic solvent and water is used as the elution mobile phase, which causes a secondary problem. After removing the low boiling solvent, the recovered portion must be dewatered overnight or several days. In this way, even if the separation process is completed, the recovery of the component of interest is delayed by several hours to several days. The latter problem can cause significant obstacles (bottlenecks) in the overall purification process when many samples are in a row.

[0006] When there are conditions that are difficult to separate, or when the separation rate is important, what is known as high performance liquid chromatography (HPLC), which is a type of elution chromatography, is selected. This HPLC technique is used for both of the following purposes: That is, it is used for both purposes as an analytical means for identifying each component and as a preliminary means for purifying and collecting these components.

[0007] In HPLC as an analytical instrument, component levels in a sample typically range from nanograms to micrograms. HPLC systems as a preliminary measure typically handle amounts of micrograms to several grams of component per separation. The HPLC system as a preliminary measure also requires a means of collecting and storing each part. This is usually done manually or automatically by simply switching the fluid flow of the system for a series of open containers.

[0008] There are drawbacks to the current HPLC as a preliminary measure. Each sample requires an elution cycle in the range of minutes to hours. Moreover, even under optimal conditions, only a small proportion of the mobile phase contains the component of interest. This can lead to a very large amount of wasted mobile phase even if the system operates normally.

An alternative separation method, called supercritical fluid chromatography (SFC), has progressed over the last decade. Carbon dioxide (CO) is typically the main component of SFC₂) Is adopted, and a highly compressible mobile phase is used. CO₂In addition, the mobile phase often contains an organic solvent modifier to adjust the polarity of the mobile phase to optimal chromatographic (separating the mixture into components) performance.

[0010] Since different levels of organic modifiers are required for rapid elution of different sample components, a common technique is to increase the organic modifier linearly to allow the mobile phase to elute. The composition is continuously changed. This technique is called the gradient elution method.

[0011] For application as analytical means, SFCs are being demonstrated to be superior in velocity and dissolving power over traditional HPLC. This is a result of the dramatic improvement in the diffusivity of the eluate in the SFC mobile phase compared to the HPLC mobile phase.

[0012] With the SFC device, separation continued to be achieved faster on the magnitude order (about 2.5 times for each rank up) compared to an HPLC device using the same separation tube (chromatographic column). The main factor (key factor) for optimizing SFC separation is the ability to independently control the flow, concentration and composition of the mobile phase throughout the separation process.

The SFC apparatus used with the gradient elution method also rebalances much more quickly than the corresponding HPLC apparatus. As a result, the device is ready to process the next sample after only a short time. The general slope range in the slope SFC technique is preferably in the range of 2% to 60% of the composition of the organic modifier.

[0014] CO₂Although the SFC device was designed to operate in the temperature and pressure range above the critical point, operating the SFC device far below the critical point is typically suppressed. It is worth noting that this is not the case. In this low range, especially when using organic modifiers, the chromatographic behavior remains superior to traditional HPLC and is often indistinguishable from true supercritical operation.

[0015] In SFC as an analytical means, once separation is performed, the highly compressed mobile phase, once detected, directs to the fluid flow through a decompression step. CO in the mobile phase during decompression₂The components are allowed to expand dramatically and return to the gas phase. Its expansion and subsequent CO₂Phase changes tend to have a dramatic cooling effect, perhaps on wastewater elements. If you're not careful, solid CO known as dry ice₂May occur and clog the wastewater. Heat is typically applied to the fluid stream to prevent this from occurring. At low flow rates in a typical analytical system, only a small amount of heat is needed.

[0016] CO in the SFC mobile phase₂Moderately heated liquid organic modifiers typically remain in the liquid phase, although the components immediately convert to a gaseous state. In general, the dissolved sample carried within the SFC system also remains in the dissolved liquid organic modifier phase.

The principle that the mere decompression of the mobile phase in the SFC separates the flow into two parts is of great importance with respect to the use of this technique in a preliminary manner. CO, which accounts for 50% to 95% of the mobile phase during normal operation₂Eliminating the gas phase of the material will significantly reduce the liquid collection volume for each component, resulting in a reduction in the amount of post-chromatographic processing to recover the separated components.

[0018] A purification technique as a second analytical means, similar to SFC, is supercritical flow extraction (SFE). Generally, in this technique, the goal is to separate one or more components of interest from the solid matrix.

[0019] SFE is a mass separation technique that does not necessarily attempt to separate the components extracted from the solid matrix individually. Secondary separation steps are typically required to determine the individual components. Nevertheless, the SFE shares a common goal with the preliminary SFC to collect and recover the dissolved components of interest from the supercritical fluid stream. As a result, a suitable collector for the preliminary SFC is also suitable for the SFE technique.

[0020] In order to extend the SFC technique as an analytical tool to serve as a preliminary SFC, some adaptation of the device is required. First, the system requires increased flow. Flow rates from 20 mL / min to 200 mL / min are suitable for separating amounts of material ranging from a few milligrams to grams. Also, a larger separation tube (column) is required. Finally, a collection system must be developed that allows at least a single portion of the fluid flow to be collected, including a sufficiently purified component of interest.

[0021] In addition, there is often an irresistible economic motivation to make it possible to collect multiple portions from a single sample. The modified system must also be able to re-initialize, either manually or automatically, quickly so that the next sample injection can be made following partial collection.

[0022] Several commercial examples of preliminary SFC equipment have been attempted that employ different levels of technology to solve collection problems. Representative samplings of these products include those proposed by Gilson, Thar, Novasep, and Pro Chrome and others.

[0023] However, none of the current instruments have succeeded in providing instruments with high recovery, high purity, and low carry-over from sample to sample. For example, some systems use the naive method of simply spraying the collection stream directly into a large bottle, the result of which is a large amount of sample loss, probably due to the formation of aerosols.

[0024] Another system uses a cyclone-like separator to separate the two streams, but does not provide rapid or automatic cleaning means to clean the separator to prevent carry-over. Such devices are typically employed for the purpose of separating large amounts of material by repetitive injections to eliminate the need for sample-to-sample cleaning steps.

[0025] In another system, a collecting solvent is used to take the sample portion into a certain volume of solvent contained in the collecting container. This technique uses a relatively large amount of dangerous preparation to collect the sample and tends to result in loss of enrichment or quality deterioration of the sample portion, subdivided sample and collection solvent composition. There may be matrix interference with the object.

An example of an SFC system is shown on the outside surrounded by frame 10 in FIG. The schematic flow path diagram is a supercritical flow chromatography (SFC) system in which separation tubes are bundled from the regulator supply to the detector. The system has a carbon dioxide supply tank 200, a line refrigerator 220, a pump 202, a regulator tank 204, and a pump 206, leading to an attenuator and pressure converter 208, a stirring column 210, and at least one bundled chromatograph. Connects to the injection valve 212 and detector 216, which connects to the graph separation tube 214.

[0027] In the SFC system, liquefied carbon dioxide gas is supplied from the cylinder 200. High-pressure piping 218 connects the carbon dioxide storage tank 200 with the carbon dioxide pump 202. This pipe may be cooled 220 before connecting to the pump 202. This system uses two HPLC type piston pumps 202 and 206. One pump 202 delivers carbon dioxide and the other pump 206 delivers a regulator 204 such as methanol. This carbon dioxide and the modifier are combined to form a modifier mixture dissolved in a supercritical flow.

[0028] The combined supercritical flow is pumped from the stirring column 210 through the transfer tube to the fixed loop injector 212 at a controlled flow rate, where the sample of interest is injected into the flow path system. To. This sample combines with the regulator compressed in the infusion valve 212 and is expelled into at least one bundled chromatographic separation tube 214. After the sample is subdivided within the separation tube 214, the elution mixture moves from the exit of the separation tube into the detector 216.

[Outline of the Invention] One of the objects of the present invention is to provide a partial collecting device for supercritical fluid diversion.

[0030] A further object of the present invention is to provide an apparatus for collecting a component obtained by subdividing a sample solution into one or more collection containers.

[0031] The present invention relates to supercritical flow chromatography, or supercritical flow extraction, and sample recovery after separation by an improved version thereof.

[0032] More specifically, the present invention expands or depressurizes a liquid phase containing a sample of a component of interest, controlling a mobile phase for separating one component from a high working pressure to an unstable low pressure. It concerns the separation under optimal conditions from the much larger gas phase after it has been done. Its controlled expansion causes phase separation between the liquid and gas phases, while at the same time strongly suppressing aerosol formation in the transfer tube.

A further object of the present invention is a single-phase fluid that is a mixture of a highly compressible or liquefied gas and a liquid organic modifier before collecting the liquid phase moiety in one or more unique collection chambers. It is to provide an apparatus and a method for separating a gas phase and a liquid phase in a transfer pipe. Collecting the liquid phase portion in this collection chamber minimizes the use and waste of liquid solvent through efficient separation of the gas and liquid phases before entering the collection chamber. This collection technique does not use additional solvents for partial collection.

[0034] The present invention provides a series of cassettes consisting of a large number of chambers for collecting and storing separated or extracted portions. Each collection cassette contains one or more collection chambers, each of which can receive a purified liquid portion.

Each chamber can accommodate a removable sample collection liner (lining). This collection liner can be individually removed, replaced, stored, cleaned, reused or discarded. One of the purposes of this collection liner is to provide a simplified means for transferring the liquid portion collected from the cassette. A second purpose of this collection liner is to provide a means of removing subsequent mutual turbidity between samples by providing an easily replaceable, clean liner for each sample in each collection room. It is to be.

[0036] In the present invention, one for a collection chamber can be used to collect a plurality of liquid phase portions from one sample in one or more collection chambers without mechanically adjusting the collection chamber seal. The above valve and sealing mechanism are controlled manually or automatically. This method allows for rapid switching between collection chambers in the event of close separation peaks in a fluid stream that separates the mixture into components.

[0037] A further object of the present invention is to facilitate manual or automatic reset of the collection system so that subsequent samples can be processed quickly. There are technical difficulties in realizing a collection system that satisfies all the goals of the analytical engineers mentioned above. The main problem is the supercritical CO of compressed liquids or mobile phases that gasify radically under atmospheric pressure.₂Concentrate around a significant expansion (typically 500 times) of the part. This change has four major negative effects on liquid phase sample collection.

[0038] First, as described above, the expanding CO₂Causes a significant temperature drop, which can form dry ice and block the system. Preliminary SFCs are much faster than their corresponding analytical systems, so a significant amount of heat must be applied to compensate for this temperature drop.

However, care must be taken: do not raise the actual temperature in the flow path system. Because the temperature rise may damage the thermally unstable component of interest. Increasing the composition of the organic regulator increases the heat capacity and CO₂The severity of this problem is alleviated with both effects of dissolving and thus preventing the formation of dry ice.

Second, CO₂As it expands, the dissolving power it had in its compressed state quickly disappears. If the component of interest is soluble, CO₂If it relies heavily on, the component of interest will lose the means of transport through the flow path system it originally had. Solid components accumulate and eventually clog the flow path, causing system failure. Again, the organic modifier component is an important factor. Because the liquid continues to dissolve the components of interest and transfer them to the collector. Care must be taken not to introduce excessive heat into the fluid stream, as organically tuned ones are also driven into the gas phase. Otherwise, the beneficial effect of transferring the solution is lost.

Third, liquid to gaseous CO in the shortest possible time after the first decompression stage.₂It is beneficial to complete the conversion to. CO while in liquid state₂Can disperse an organic modifier containing a component of interest, even if the concentration is not sufficient to have outstanding dissolving power. This dispersion can have the effect of stirring the components efficiently separated by SFC treatment prior to depressurization. CO₂The faster the conversion, the less the deterioration of the function of separating into components.

[0042] Liquid CO₂Two factors seem to dominate in controlling the evaporation capacity of. : a) Sufficient heat transfer between the heat source and the flowing liquid, and b) CO in the heated section₂Staying time.

[0043] The first factor can be influenced in a positive direction by selecting a highly conductive material such as copper when manufacturing the heater. Ensuring good thermal contact between the heater and the transfer tube made of thin walls also facilitates heat transfer to the fluid stream.

[0044] The residence time of the depressurized fluid can be controlled by stepwise reducing the pressure by connecting one or more suppressors in series with the transfer tube. Increasing the back pressure reduces the linear velocity of the two-phase fluid in the heater. As long as back pressure is generated by these suppressors, it does not interfere with the SFC concentration control in the high-pressure separation region, and a large adjustment allowance for heat transfer optimization is possible.

Fourth, due to expansion, the linear velocity of the decompressed fluid in the transfer tube increases dramatically. The residual liquid in this system is moved along the flow path, largely by the shearing force of the expanding gas. This spiral environment is ideal for the formation of aerosols, where the microbubbles of the modifier liquid are carried away as "mist" into the gas phase. It is a finding from this study that aerosol formation in transfer tubes can be almost completely controlled by proper temperature control of the expanding two-phase system. At lower temperatures, aerosol formation is a greater problem. Corresponding low level CO₂It is a surprising finding from this study that higher levels of organic modifiers containing ingredients require higher temperature levels to prevent the formation of visible aerosols.

[0046] As a preferred exemplary embodiment, the SFC collection system consists of a moderately restrained, thermally controlled stainless steel transfer tube. This transfer tube extends from the pressure control element of the SFC chromatograph to the multi-port distribution valve, which leads to a variety of streams that connect directly to a separate collection chamber row or a common effluent container with vents. It extends to the road.

[0047] CO₂The first separation of the liquid phase sample from the gas takes place immediately at the first decompression site within the back pressure controller of the SFC or SFE device. Solid CO by performing downstream suppression₂Liquid CO while maintaining a minimum back pressure sufficient to prevent the formation of₂Is present in the transfer tube.

[0048] Remaining CO from the organic modifier₂Evaporation and separation of is carried out in a stainless steel transfer tube before entering the cassette. This is accomplished by contacting the transfer tube with a series of heaters designed to optimize heat transfer to the fluid. Ideally, this series of heaters will generate enough energy in the liquid CO of the fluid.₂Transfer to the part and its liquid CO₂The part is completely evaporated and the temperature of the fluid is raised sufficiently to prevent external freezing of the transfer tube. Since the rate of heat transfer is time dependent, it is beneficial to slow down the speed of the fluid in a series of heaters.

CO in the first heating zone₂During the evaporation process of₂There is considerable separation between the liquid and the liquid regulator. But for some reason, pure CO₂And separation into liquid modifiers are not realized.

[0050] First, typically small amounts of organic modifiers also evaporate into a gaseous state. The degree of evaporation largely depends on the absolute temperature of the fluid in the transfer tube. Although the recovery of the liquid phase decreases as the organic modifier evaporates, the recovery of the dissolved component of interest does not necessarily decrease because the organic modifier converts to vapor and the boiling point is typically not very low.

Second, a part of CO₂Remains dissolved in the organic liquid. Both temperature and pressure are residual CO₂Determine the amount of. CO at high temperature₂Solubility is reduced, but at high pressure CO₂Increases the solubility of.

Aerosol formation in the liquid phase is a common problem in SFC sample collection and is a major cause of wear on the organic liquid phase, including dissolved components of interest. Aerosol production is reduced at high temperatures. The composition of the separated phases is also a factor. Higher temperatures are required to remove aerosols in fluids containing higher concentrations of organic liquid phase. An additional heating zone is used to regulate the fluid temperature for aerosol control. In addition, the heater allows precise levels of fluid temperature control prior to collection in a pressurized collection chamber.

[0053] As described above, the secondary effects are as follows. High level of temperature control, melted CO₂Can reduce the concentration of CO, resulting in uncontrolled or explosive CO when pressure is removed from the collection chamber.₂The possibility of gas generation can be reduced.

[0054] Following the adjusting heater, a valve system is used to continuously divert the two-phase fluid flow to abandoned flow or to one of the collection chambers in the collection cassette. This valve system consists of one or more valves and an electrical controller.

The system is designed for rapid response to manual or automatic start / stop signals. Typically, this signal is based on the results of detecting a component of interest coming out of a high pressure fluid system.

The start signal is emitted when the first component is detected, while the stop signal is emitted when the detection is lost. The effect of the start signal is to divert the fluid flow to one of the first unused collection chambers in the cassette. The effect of the stop signal is to switch the course of the fluid flow to abandoned flow.

Another possible form of start / stop signal is based on the timetable rather than the physical detection of the components. The controller may also have a form that limits the access time or the permissible flow rate to each room. In addition, if more parts are desired than there are currently available collection chambers, this controller can allow or prevent the system from returning to the first chamber.

A collection cassette is a device that can be resealed, including one or more hollow collection chambers with an open top. As a preferred representative embodiment according to the present invention, each chamber can hold a removable inert liner. The liner collects a portion of the initial sample dissolved in a liquid solvent base. A cassette as a preferred representative embodiment has four chambers as a case for accommodating four glass test tubes, which functions as a chamber liner. The number of chambers in a cassette can be increased or decreased regardless of performance. Each glass test tube can hold a sample portion separated from the high pressure fluid stream up to its capacity.

[0059] As a preferred embodiment, the sample portion is collected in one chamber of the cassette at one time. Two-phase fluid enters a chamber from the valve system via a transfer tube. The tip of the transfer tube is preferably located at right angles to the inner wall of the collection tube and slightly downwards, usually 45 degrees or less from the horizontal.

[0060] A spring wire for a guide is wound around a transfer tube and suspended inside the test tube. This spring wire is bent so as to protrude from the transfer tube and acts as a guide when the transfer tube descends into the glass tube. When the transfer tube is successfully inserted into the glass test tube, the bent portion of the spring wire hits the outer peripheral edge of the open end of the glass test tube. As the transfer tube continues to descend into the glass tube, the spring wire exerts pressure on the inner surface of the glass test tube, pushing the transfer tube toward the opposite side of the glass tube. As a result, the tip of the angled transfer tube is pressed against the inner surface of the glass test tube.

[0061] Organic liquid and CO₂Both gases descend a spiral passage along the inner wall to the bottom of the collecting liner. The liquid is collected at this point and begins to fill the liner. That CO₂The gas continues to rise through the passageway in the center of the liner towards the exhaust holes in the collection room. Suppressive transfer pipes attached to the exhaust holes, both inside and outside the collection liner, CO₂Gas causes high pressure in the collection room. The degree to which back pressure is applied to the collection room is the CO of the initial mobile phase.₂It is roughly proportional to the composition.

[0062] Higher pressure in the collection room is the CO that enters the collection room.₂Helps slow down the speed of gas. In other words, CO at the bottom of the liner₂It reduces the magnitude of the shear force that occurs between the gas and the collected liquid. The smaller the shear force, the less likely the collected liquid will be an aerosol and the less likely it will be removed from the collection tube with the active gas. A similar effect can be obtained by properly angling the injection transfer tube relative to the wall of the collection tube. The closer and smaller the angle of the transfer tube is, the less swirls are observed on the liquid surface. However, sufficient angle must be provided to ensure that most of the effluent points the liner wall downwards rather than upwards.

[0063] The suppression of aerosol formation in the collected liquid portion is a combination of two effects: back pressure and injection angle. Successful optimization of these effects depends on how close the infusion tube can be placed to the collection fluid and, as a result, how high the liner is filled before sample loss becomes an issue. It's done. When the flow to the collecting room stops, the room is decompressed. Once the sample chamber is decompressed, the liner can be removed by opening the top lid of the cassette.

[0064] Collecting parts into a disposable liner in a collection room can be automated using a robot. In the automated system, glass test tubes can be quickly replaced into and out of the collection chamber, and a large number of glass tubes for replacement can be prepared for long-term unmanned operation. A programmable robot automatically arranges the cassettes in a fixed order at sample injection intervals, resulting in faster processing and less room for error. This automated system can collect thousands of orders per month.

The automation system is housed in a laboratory grade case. The system includes a robotic arm, an inventory of glass test tubes placed upwards in the rack, and an automated version of the cassette assembly. In addition, the system may include sufficient probes, valves, and sample containers to achieve automatic injection of unseparated samples into the chromatography or extraction system.

The collection cassette and its automation mechanism are designed for rapid sample collection and the shortest downtime of the chamber liner replacement interval. The cassette in the preferred embodiment has two rows, each with four collection chambers. A lid is placed above the row of collection chambers in the cassette. The lid has four partially recessed holes corresponding to the four collection chambers in the cassette. The lid is mounted on the base of the case and is raised and lowered by the movement of pneumatic actuators located at both ends of the lid in the longitudinal direction. As each actuator simultaneously lowers the lid onto the collection cassette, the top edge of each chamber engages the bottom edge of the lid, which corresponds to the respective rim of the partially recessed hole.

[0067] The lid and the collection chamber are engaged to form a pressure-sealed seal in each chamber in the standby state for partial sample collection. The lid has a transfer tube and a drainage tube that penetrate each recessed hole, corresponding to each collection chamber. When the lid is lowered onto the cassette, each tube pair (transfer tube and drainage tube) enters the test tube.

A spring wire wound around the injection transfer tube guides the injection transfer tube into the glass test tube. The tip of the angled transfer tube is pressed against the inner surface of the test tube. When the lid seals the rows of the collection chamber, the valve system distributes the flow convection, including the gas and liquid phases, from the sample subdivision process to the chamber liner.

[0069] When all the test tubes in the pressured cassette row are filled and depressurized, the lid lifts away from the cassette. The cassette then moves laterally or reciprocates, and the empty collection room liner moves under the lid, replacing the previous row. The cassette is constrained to reciprocate laterally along the passageway on the case base.

[0070] The lid is lowered and engaged with a new collection chamber row, resulting in the preparation of a test tube to receive the sample portion. On the other hand, the previous row of collection chamber liner glass test tubes, including the liquid portion, is removed from the collection chamber and moved by a robotic arm to an empty space in the storage tray.

[0071] In summary, in a preferred embodiment, the sample is dissolved in a minimal volume of modifier solution and collected in a liner that can be removed and reused. Excellent separation of near-supercritical elution fluids was obtained by adjusting the flow rate, velocity, temperature, and pressure in the system. A collection efficiency of 98% of the injected sample components was achieved.

In this process, a cassette with a pressurized collection chamber and a disposable liner minimizes the amount of additional solvent used for collection and cleaning in the laboratory, which is economical and environmentally friendly. .. Laboratories and laboratories that require sample purity while maximizing yield and minimizing waste will benefit from this invention. Thousands of large-scale sample subdivisions and collections per month can be achieved in this representative example.

[0073] In order to better understand the properties of the present invention, the following drawings will be referred to and described in detail. Here, similar elements are given similar reference numbers.

[Example] In FIG. 1, the peripheral line 10 shows a preferred embodiment of the present invention. Unless otherwise specified below, in the system of the present invention, a highly compressed fluid flow (CO) from the pump system.₂A positively adjusted flow) shall be received at a flow rate of 20 to 100 mL / min. Depending on the embodiment, the flow velocity may be less than or greater than this by adjusting the system hardware and flow parameters.

[0075] In one embodiment, the SFC collection system has a temperature controlled transfer tube 12, which extends from the back pressure regulator 14 to the multi-port distribution valve 22 for further collection. It extends into various channels through the chamber 32 or directly into the common waste liquid container 26.

[0076] The expansion waste liquid leaving the regulator 14 has a flow velocity of about 2 to 5 times that of the upstream side, and the back pressure is about 20 to 40 bar. Fluctuations in swelling occur by changing the concentration of the adjusting solvent over the process of 2.5-50% separation.

[0077] The initial separation of the liquid phase from carbon dioxide occurs at the time of initial decomposition within the back pressure regulator 14 of the SFC or SFE system. Liquid CO by suppressing the downstream side₂Is present in the transfer tube 12, but the individual CO₂Sufficient minimum back pressure is maintained to prevent the formation of. CO₂The degree of evaporation is a function of the heat transfer and downstream flow suppression that can be adopted, which limits the amount of expansion of the decompressed fluid. CO released due to pressure drop in regulator 14₂A portion of the fluid evaporates, typically lowering the temperature of the ejected fluid.

CO from organic modifiers₂Further separation and evaporation occur in the transfer tube 12 extending between the regulator 14 and the cassette 24. Transfer pipe 12 is a two-phase CO₂It contains a fluid flow with the regulator and is heated by a series of heaters 18 and 20, which optimize the heat transfer to the two-phase fluid in the fluid flow. These heaters are the liquid CO of the discharging fluid₂Ideally, the fluid should be heated to a temperature sufficient to transfer sufficient energy to the portion to allow its complete evaporation and prevent freezing to the outside of the transfer tube 12.

CO in the first heating region₂Gas phase CO during evaporation process₂And a significant separation between the liquid phase modifiers. But pure CO₂And separation into pure modifiers is never observed. The degree of evaporation largely depends on the absolute temperature of the fluid in the transfer tube 12.

[0080] Evaporation of the organic modifier results in reduced liquid phase circulation when it reaches the collection cassette, but does not necessarily have a component of interest (generally not having a sufficiently low evaporation point to transform into the above). ) Is not reduced. Also CO₂A part of is dissolved and remains in the organic modifier. Temperature and pressure are residual CO₂Determine the amount of. The higher the temperature, the more CO₂Solubility is reduced, but the higher the pressure, the more CO₂Increases the solubility of.

[0081] Gas phase CO in a narrow transfer pipe₂Turbulence creates a strong shear force that propels the fluid down the wall of transfer tube 12. This turbulence often produces droplets on the surface of the fluid that break up from the liquid and are trapped in the gas phase of the fluid rapidly descending the transfer tube 12. Such an effect is called aerosol formation or atomization.

[0082] A plurality of heaters may be connected in succession to heat the waste liquid fluid. In the embodiment of FIG. 1, the evaporation heater 18 and the trim heater 20 are continuously arranged after the regulator 14. The heater 18 is heated by a cartridge heater of an appropriate size and controlled by an appropriate controller. The transfer tube 12 is tightly wound around the heating mechanism to optimize thermal contact.

[0083] The waste liquid fluid is heated within the control temperature (about 5 to 50 ° C.) of the heater 18 to prevent damage to the heat-sensitive composite. The purpose is to pass the heater 18 through the vortex to C0₂Is to be boiled from the waste fluid. To complete the required heat transfer, the two-phase fluid in the transfer tube 12 enters the collection heat exchanger, the trim heater 20. It is generally desirable to set this trim heater above the set point of heater 18. This heater 20 not only suppresses aerosol formation in the transfer tube 12, but also CO to the liquid phase.₂It also controls the solubility of.

[0084] The velocity of the fluid passing through the heaters 18 and 20 in the transfer pipe 12 should be low. For this purpose, a constraint orifice or a small diameter portion may be provided immediately downstream of the first heater group. The waste fluid leaving the heater 18 enters the current controller 16. This current control device applies high back pressure to the heater 18 to slow down the flow and liquid phase CO.₂Increases contact time.

[0085] In addition, the back pressure is sufficiently high due to the current control device 16, and the liquid phase CO₂Is a solid phase CO known as dry ice₂Is prevented from forming. This flow control increases the back pressure in the heating region and reduces the amount of gas expansion. The fluid may be slowed down after all heating, but CO causes uncontrolled cooling of the fluid in the transfer tube.₂Unable to control the final expansion of. As a result, the ability to positively suppress the formation of aerosols is impaired.

[0086] After leaving the trim heater 20, the transfer pipe 12 is connected to the common port of the distribution valve 22. It is desirable that the distribution valve 22 is a multi-port selection valve. As the waste fluid passes through the distribution valve 22, the gas and liquid phases are directed to the collection cassette 24 or the waste container 26. The outlet port of the distribution valve 22 is connected to a plurality of transfer pipes 28.

[0087] The transfer tube 28 is connected to the collection chamber 32 in the cassette 24 through the cassette lid 30. The transfer pipe 28 is airtightly and pressure-resistant to the cassette lid 30. The other ports of the distribution valve 22 are connected to the transfer pipe 34. The distribution valve may be provided with a plurality of sorting valves and connected to the transfer pipe 12. Each valve port is connected to an individual collection chamber 32, and the sorting valve is connected to a waste liquid transfer pipe 34.

The transfer tube 28 that has entered the collection chamber 32 is inserted into the bin 36. The liquid phase is trapped in this bottle 36, while the gas phase escapes from the collection chamber 32 through the discharge pipe 42. The gas in the discharge pipe 40 flows at high pressure. The drain pipe 40 from the cassette 24 flows through the pressure relief switch 42 to prevent the cassette and upstream components from damage due to overpressure due to system malfunction.

[0089] Also, in the embodiment of FIG. 2, the cassette 24 has four different collection chambers 32. However, the cassette 24 may include one or more individual collection chambers 32. Each collection chamber 32 may be a closed system as a final separation point between the liquid phase and the gas phase. The collection chamber 32 is a hollow cylinder made of high-strength transparent plastic that allows the separation and collection process to be visible.

The collection chamber 32 is made of stainless steel or a suitable laboratory grade material. The collection chamber 32 is arranged in a parallel and upright position in the cassette 24. Each collection room 32 is closed at the upper and lower ends in frames 44 and 46. Each collection chamber 32 has an open end surrounded by the upper frame 44, and the lower end is partially embedded in the lower frame 46. Communication between the liquid phase and the gas phase between the collection chambers 32 is prohibited by the seal 48, which is loaded into the groove 50 at the upper opening end of each collection chamber 32.

Each collection chamber 32 houses a replaceable liner. The bottle 36 functions as a liner and is arranged upright in each collection room 32. The closed lower end of the bottle 36 rests on the base of the collecting room 32 and can be easily replaced. Once inserted, the top edge of the bin 36 should be lower than the height of the collection chamber 32 and the internal recess 60 (FIG. 3) when the lid and cassette 24 are engaged. The bin 36 and the collection chamber 32 are a single pressurization system that communicates with each other through the upper end of the collection chamber 32.

The bin 36 acts as a disposable liner for the collection chamber 32 and captures the liquid phase 38 separated from the fluid stream. The inside of the bottle 36 and the annular space of the collection chamber 32 surrounding the bottle are balanced at the same pressure and are in the range of about 20-100 psig during the separation process up to a flow rate of 50 ml / min. With such a configuration, it is possible to partially collect a sample at high pressure in the collection chamber 32 without the risk of the bottle 36 being destroyed.

[0093] The cassette 24 shown in FIG. 2 has a square frame for fixing four upright collecting chambers 32. The frames 44 and 46 properly position the collecting chamber 32. Two square end pieces 52 are attached to the upper and lower ends of the frame. Each end piece 52 is a metal plate and is fixed by a screw 54. A latch 56 provided at the upper end of the end piece 54 fixes the cassette lid 30 to the upper end of the cassette 24. The lid 30 can be manually removed during sample injection, which allows quick access to and removal of bin 36.

[0094] As shown in FIG. 1, a transfer pipe or an orifice 33 is provided at the bottom of each collection chamber 32, and extends through the bottom and the frame 46. The orifice 33 allows the liquid phase to be removed without the need to open or decompress the collection chamber 32. The orifice 33 also facilitates drainage cleaning of the collection chamber 32 during maintenance of the cassette 24.

[0095] FIGS. 3A and 3B show the head and bottom of the cassette lid 30. The lid 30 has four sets of three holes 58 in a triangular shape, and each hole is located directly above each collection chamber 32 when the lid 30 engages with the cassette 24. On the bottom surface of the lid 30, there is a hole 60 located directly above the collection chamber 32 when the lid 30 and the cassette 24 are engaged.

[0096] The diameter of the hole 60 is designed to be slightly smaller than the diameter of the collection chamber 32. When the lid 30 is secured to the cassette as shown in FIG. 2, the perimeter of the hole 60 is located completely inside the seal 48. The holes 60 allow the bin 36 to stand higher than the top surface of the frame 44 so that the bin 36 can be removed without access to the collection chamber 32. This avoids contamination of the next sample.

[0097] A pin 62 shown in FIG. 3 is formed on the upper surface of the cassette 24 so as to guide the lid 30 and the cassette 24 together at the time of engagement. The hole 63 in the lid 30 receives the pin 62 from the cassette 24. A catch 64 that catches the latch 56 is attached to the long end of the lid.

[0098] The transfer pipe 28 conveys the liquid phase and the gas phase to the bottle 36. Each transfer pipe 28 is fitted into the hole 58 of the lid 30 and inserted into the bottle 36. This mating provides an airtight connection that can withstand the pressure in the SFC system. The transfer pipe 66 directs the waste liquid into the bottle 36, and the transfer pipe 68 constitutes an escape route for the pressurized gas, whereby the gas exits the collection chamber 32 and heads for the waste liquid container 26.

[0099] In a preferred embodiment, a small amount is collected at one time in the collection chamber 32 of the cassette 24. In the process of reducing the amount, both the liquid phase and the gas phase are discharged to the bottle 36, where the final separation process occurs. CO in the room by pressurizing the collecting room 32₂Slows down. This will result in CO₂Shear force between the and the liquid collected at the bottom of the bottle 36 is reduced. When the shear force is low, the collected liquid is less likely to become an aerosol and is removed from the bottle 36 together with the exhaust gas.

[0100] A similar effect can be obtained by angling the transfer pipe on the inflow side to the bin 36. The closer the angle of the tube 66 is to the horizontal, the lower the turbulence observed on the liquid surface. However, it must be angled enough to point the bottle 36 over the wall downward rather than upward.

[0101] The two-phase effluent fluid enters the collection chamber 32 from the distribution valve 22 via the transfer pipe 28. As shown in FIGS. 8A, 8B, the tip of the transfer tube 66 is preferably a probe tangential to the bin 36 liner wall and is slightly inclined downwards (usually less than 45 degrees to horizontal). is there). A guide spring 70 is attached to the transfer pipe 66, and the transfer pipe 66 is curved so as to be separated from the transfer pipe 66. The spring 70 functions as a guide when the transfer pipe 66 descends into the bin 36.

[0102] When the transfer tube 66 is properly inserted into the bin 36, the curved portion of the spring 70 comes into contact with the peripheral edge of the bin 36. When the transfer tube 66 extends into the bin 36, the spring 70 presses on the inner surface of the bin 36, pushing the probe toward the other end of the bin 36. As a result, the inclined tip of the transfer pipe 66 is pressed against the inner wall of the bin 36.

The spring 70 is extruded from an inert material and does not chemically interfere with the sample collected in the bottle 36. It is desirable that the transfer pipe 66 of the transfer pipe 28 is a stainless steel probe attached to the lid 30. A larger OD Teflon® tube is fitted at the end of the metal portion of the transfer tube 66 to prevent damage to the inner wall of the bin 36.

[0104] Organic liquids and CO₂Flows to the bottom of bin 36 along a spiral descending path. The liquid phase gathers at this point and begins to fill bin 36. CO₂The gas flows through the upper end of the collection chamber 32 to the center of the bin 36 and goes to the exhaust section. CO by the flow control pipe 72 attached to the exhaust part₂The gas pressurizes the collection chamber 32 inside and around the bottle 36. Indoor back pressure is the initial movable phase CO₂It is almost proportional to the composition of.

Back pressure and delivery angle synergistically reduce aerosol formation in the collected liquid portion. By optimizing these effects, it is possible to determine how close the transfer tube 66 is to the collected liquid. This also determines how high the bottle 36 will be filled before sample loss becomes a problem. When the flow to the collecting room 32 is stopped, the room is decompressed. Once the collection chamber 32 is decompressed, the bottle 36 containing the liquid phase is removed by opening the lid 30 of the cassette 24.

[0106] The transfer pipe 72 from the collecting chamber 32 is connected to the switch 42 to hold the pressure in the collecting chamber 32. This switch 42 is CO₂Increase the pressure on the upstream side to 20 to 100 psig according to the flow velocity of. Each transfer tube 72 passes through a pressure switch 78 to avoid overpressure and breakage. The pressure in each room is visually monitored by a pressure gauge 76 screwed into the lid. Transfer pipe 72 points to waste liquid container 26, where CO₂Is exhausted. For safety, waste liquid, sample and exhaust CO₂Must not leak into the surrounding air. In addition, the liquid and gas in the system are kept tightly closed and guided to the waste liquid container 26 or the like to maximize safety.

[0107] The volume of liquid phase 38 captured in bin 36 is controlled manually or automatically. In a preferred embodiment of the distribution valve 22, this control is performed by one or more valves and an electronic controller. The distribution valve 22 is designed to respond quickly to manual or automatic start / end signals.

This signal is obtained from the detection of components coming out of the high pressure flow system. The start signal is generated by the initial detection of the component, and the end signal is generated when the detection is lost. When the termination signal is given, the fluid flow is directed to the waste container 26 or the collection chamber 32. Also, the start / end signals may be based on the timetable rather than the physical detection of the components.

[0109] The controller may constrain the permissible access time and the ruins that have flowed to the individual collection chambers 32. In addition, if more than is present in the available collection room 32 is desired, the controller can also prevent the system from returning to the original collection room 32 and circulating.

[0110] Figures 4 to 7 show examples of changes in cassettes and systems. This is an automatic system using a robot arm 80, in which the bottle 36 is replaced by the arm after filling the sample. This allows the bottle 36 to be quickly filled and replaced over a long period of time. The supply tray 86 of the clean bin 36, which acts as a liner, is located in the collection chamber 32.

The robot arm 80 is controlled to replace one or more bins 36 from the row of collection chambers 32 in the cassette 84 with the bins 36 from the new supply tray 86. Further, the arm 80 replaces the bin 36 in the first row of the cassette 84, and the bin 36 in the second row is automatically positioned. By being robotized in this way, it is possible to minimize the replacement time of the bottle 36, speed up the collection of samples, and collect more samples.

[0112] Figures 4 and 5 show other examples of automated SFC sample collection systems. The elements of the system are partially located on the base 88 in the case 82. The pedestal 88 is supported by adjustable legs 90 distributed around it, which adjusts the height of the pedestal.

The supply of clean bins 36 is housed in rack 86 on base 88. Each bin 36 is held upright and secured to the rack 86 by a molded support. Each rack 86 is composed of a circular portion tangentially attached to an adjacent portion, and a molded support loosely secures the bin 36.

The bins 36 are held equidistant from each other, providing a space in which the gripping jaws 92 on the arm 80 can grip the individual bins 36 without interference. In addition, this space prevents damage when moving and replacing the rack 86. Only two racks 86 are shown, but the number can be expanded as appropriate.

[0115] The cassette 84 may be provided on a raised base 88. The plurality of chambers of this cassette are designed to move laterally, and this movement is controlled by a pneumatic actuator 96. This cassette 84 is called a "shuttle cassette" and is illustrated in FIGS. 6 and 7.

[0116] The shuttle cassette 84 is configured in the same manner as in the embodiment including the rows of collection chambers 102. The cassette 84 has upper and lower square frames 98, 100, which support a plurality of rows of upright collection chambers 102. Two rows of collection rooms are supported, with four collection rooms in each row. The size of the cassette 84 can be changed as appropriate to increase or decrease the number of rows and the number of collection rooms per row. In FIG. 10, there are three rows of collecting rooms 102.

[0117] Square plates 104 are provided at both ends of the cassette 84, and the plates 104 are fixed to the upper and lower frames 98 and 100 by screws 106. The cassette may have a fixed structure, but the disassembled type facilitates cleaning and replacement of damaged parts.

[0118] The collection chamber 102 is made of high-strength transparent plastic so that the collection process in the collection chamber 102 can be visually monitored. The collection chamber 102 may be formed of a high-strength material such as stainless steel that is compatible with the SFC parameters. Each collection room 102 is located within the lower frame 100.

[0119] The upper frame 98 is fixed near the upper end of the opening of each collection chamber 102. Each collection chamber 102 extends over the top surface of the cassette 84, but is separated by a distance enough to seal the collection chamber 102 with the lid 108. The bottle 36 is inserted into each collection chamber 102 to function as a liner.

[0120] The cassette 84 is configured to be movable laterally on the base. As shown in FIG. 7, the lower frame 100 or the base has a horizontal hole 110, which extends orthogonally to the opening side of the cassette. An actuator 96 is provided on the base so as to be abutted against the cassette 84, to which a rod 94 or a control arm is attached.

[0121] The rod 94 is made of a rigid material such as stainless steel, is inserted into the hole 110 of the cassette 84, and is fixed to the frame 100. Actuator 96 laterally moves cassette 84 in response to commands from a programmable control system. As shown in FIG. 7, the roller 112 may be attached to the frame 100 and guided by the groove of the base 88 of the case.

[0122] Since the track not only causes the cassette 84 to move, but also removes from the arm 94 and actuator 96 the tension during the alignment movement of the cassette 84 due to the uneven friction of the rollers or the unevenness of the surface of the base 88. is there. In addition, a guide track in the track mounted on the ball bearing can be used to cause lateral motion.

[0123] In FIGS. 6 and 7, the lid 108 of the cassette 84 is automatically controlled, and after the cassette is moved directly under the lid 108 by the actuator 96, it engages with the row of the collection chamber 102. The lid 108 may be made of stainless steel.

However, high density plastics or materials of similar composition and composition are also effective. The lid 108 has a hole 114 parallel to the vertical axis. Two nuts 118 on the top and bottom of the lid secure the lid to the rod 116. The lid 108 can only be moved up and down.

[0125] The movement of each rod 116 is controlled by an actuator 120 attached to a base 88 of the case. The equator 120 synchronously moves the rod 116 beyond the castle gen, and raises and lowers the lid 108 with respect to the collection chamber 102 in the cassette 84.

[0126] In FIG. 7, the lid 108 is raised above the cassette 84 before engaging. The bottom surface of the lid 108 is perforated with four holes 122 corresponding to the four collection chambers 102, and when the lid 108 is lowered onto the cassette 84 by an actuator, each collection chamber 102 is partially inserted into the holes 122. Will be inserted. The lid 108 stops at a programmed point and the circular edge of each hole 122 engages and seals on the flat top surface of the frame 98.

The diameter of each hole 122 in the lid 108 is larger than the diameter of the collecting chamber 102. As the lid 108 descends onto the cassette 84, the holes 122 align with the top of the opening in the collection chamber 102. The large diameter of the hole 122 completely surrounds the upper end of the opening of the collection chamber 102. An appropriate O-ring or the like is placed around the collection chamber 102 between the upper end of the cassette 84 and the lid 108 to provide an airtight and pressure-resistant seal when both elements engage.

Positioning pins 124 are arranged on both sides of the row of collection chambers 102 on the top surface 98 of the cassette 84. These pins 124 are hemispherical on the top surface of the cassette 84, thereby preventing misalignment of the cassette 84 with respect to the lid 108. When the lid 108 engages the cassette 84, these positioning pins engage the holes 126 in the lid.

The collection chamber 102 is the place where the final separation of the liquid and gas phases takes place. Communication between the two phases between the collection chambers 102 is prohibited by a lid 108, which automatically seals each collection chamber airtightly as it descends into a row of collection chambers in the transfer tube 84. As with the cassette, each collection chamber in the cassette 84 holds a bin 36 for capturing the liquid phase.

This bin 36 is a standard laboratory test tube. The closed bottom of the bottle 36 rests on the base of each collection chamber 102, which rests on the lower frame. Bin 36 and collection room 102 communicate as a single pressurization system. After the lid 108 engages the cassette 84 as shown in FIG. 8B, the open end of the bin 36 is located at the upper end of the hole 122.

[0131] The internal pressure of the parts surrounding the bottles of the bottle 36 and the collection chamber 102 is in the range of 20 to 100 psig during the collection process. This balances the pressure inside and outside the bottle 36 and enables collection in the collection room at high pressure using standard low pressure glass or plastic bottles.

As shown in FIG. 7, the closed lower end of each collection chamber 102 has a sample discharge port 128, which penetrates the lower frame 100. A plug is inserted in each port 128 while the cassette 84 is in use. This port 128 makes it possible to remove the liquid phase collected directly in the collection chamber 102 without using a liner. By discharging the liquid phase through the port, the liquid phase can be collected without detaching the lid 104 from the cassette 84. The liquid phase may be evaporated from the collection chamber 102 under pressure, or may be delivered by gravity after the collection chamber is decompressed.

[0133] The transfer pipe 66 and the transfer pipe 68 transfer the liquid phase and the gas phase between the cassette 84 and the external transfer pipe, and are shown in FIGS. 6 to 7. These penetrate the lid 108 and are made of high pressure stainless steel or the like. The transfer pipe 66 carries the gas phase and the liquid phase to the collection chamber 102 under high pressure.

[0134] The transfer pipe 68 carries the separated gas phase to the waste liquid container 26 and discharges it. The lid 108 has four sets of triangular three holes 134 penetrating the lid and corresponds to the collection chamber 102 when the lid engages the cassette 84.

[0135] The pressure in the collection chamber can be measured by a pressure gauge 76 screwed from the top of the lid 108 into any of the holes 134 in addition to the transfer tube. Transfer tubes 66, 68 and pressure gauge 76 are all airtightly fitted to withstand the pressure generated within the SFC system. When the lid 108 engages the cassette 84, the transfer tubes 66, 68 are inserted into the bin 36 below the lid 108.

The tip of the transfer tube 66 is set at an angle of less than 45 degrees and is wound with a spring 70. This spring is curved along the vertical section. The spring 70 applies pressure to the inner wall of the bin 36 to incline the transfer pipe 66 in the bin 36. As a result, the open end of the transfer pipe 66 is tangential to the opposite inner wall of the bin 36.

[0137] It is desirable that the transfer pipe 66 has such a structure because the liquid phase leaving the transfer pipe 66 comes into contact with the side wall of the bin 36 and descends in a spiral shape along the inner wall of the bin 36. This swirling motion causes the final separation from the gas phase to the liquid phase, and prevents the sample from being re-engaged from the liquid phase to the gas phase and carried to the waste liquid container 26 for loss.

[0138] A Cartesian or three-dimensional robot arm may be programmed to move the bin between the supply rack and the collection room. In FIGS. 4 and 5, the robot arm 80 is attached to the wall of the case 82 near the cassette 84. The host PC or microprocessor issues a positioning command and moves the arm to control the automatic function. Joe 92 on the arm 80 grabs bin 36 and places it from rack 86 to cassette 84.

[0139] This jaw 92 holds a bottle 36 of a specific diameter at a specific position in the case 82. In the embodiment shown in FIG. 5, the robot arm 80 causes the jaw 92 to hold the bin 36 and move it between the cassette 84 and the rack 86. To increase the volume of the exchanged bins 36, the jaws 92 may hold multiple bins 36, may have multiple jaws on one arm 80, and may have multiple jaws. An arm may be used.

[0140] The movement of the arm 80 is matched with the movement of the cassette 84. When the row of collection chambers 102 in the cassette 84 engages the lid 108, the robot arm 80 replaces the bin 36 filled with the collected samples in the cassette with a new bin 36 from the rack 86. When one row of bins 36 in the cassette 84 is replaced and the row of bins 36 below the lid 108 captures the liquid phase, the controller signals the pneumatic actuator 120 that controls the lid to pull the lid 108 from the cassette 84. Let go. The lateral control 96 of the cassette 84 is then signaled to move the cassette so that the collection chamber 102 containing the fresh bottle 36 is placed directly under the lid 108 prior to engagement.

The actuator 120 is then signaled to engage the lid 108 again with the cassette 84, which prepares the collection chamber to pick up the liquid phase. The robot arm 80 then grabs the bin 36 containing the liquid phase from the collection chamber 102 and places it in the rack 86. The arm 80 then replaces the new bin 36 until each collection room 102 is full. This process continues across sample lengths or until there are no more bins from rack 86.

[0142] In the case of the cassette shown in FIG. 9, the integrated cassette 140 is composed of a plurality of rows of grid-shaped wells 144 formed in the same manner as the dropping tray. Since the foot of the cassette 140 is small, the number of collection chambers above the cassette 84 can be increased. The cassette 140 also functions as a storage tray for the collected liquid phase.

[0143] Thus, the time and cost of the replacement process is saved by eliminating the hassle of replacing the bin 36 or bringing the liner from a separate storage area. By modifying the mechanism of the lid 108 and the automatic collection system, the integrated cassette 140 acts as a simple collection cassette and storage tray by itself, allowing quick collection of samples without changing bottles 36 between each sample injection. ..

The robot arm 80 can replace the entire integrated cassette 140 after the collection process is complete or after the well 144 contains the desired amount of liquid phase. When multiple cassettes 140 are stored in an automated collection system, many harvested samples can be given during the automated process. In the preferred collection cassette, collection chamber rows housed in deep wells are used in the pharmaceutical industry.

[0145] According to such a type, not only more collection chambers are included per domain, but also those collection chambers are easy to accumulate, so that the storage density is improved and the sample storage capacity is improved. The integration cassette 140 is made of a high-strength material such as plastic, resin or stainless steel.

[0146] The integration cassette 140 can also include replaceable bins 36 in wells 144 or can be liner-free, allowing collection samples to be placed directly in wells 144 for rapid sample collection. It is advantageous. After the well 144 is filled with the liquid phase, the cassette can be replaced as a whole.

[0147] When a cassette tray is used, it is the same as that in FIG. 4, but there are some changes. That is, instead of the rack 86, a space for the cassette tray is provided, the dimensions of the lid 108 and the controller 120 and the transfer tubes 66 and 68 are devised, and the dimensions of the lateral controller 96 for the tray 140 are devised and collected. The collection chambers 144 are switched inside, and the robot arm 80 is changed to replace the filled tray 140 with a new tray from the supply area. Also, the movable lid 108 is connected to the robot arm 80 so that it engages with each row of collection chambers in the supply rack without moving the tray.

[Effects of the Invention] As described above, the sample collection system of the present invention has several advantages. For example, simplified pre-SFC sample collection is possible, only the part of interest can be collected from the injected sample, and the washed sample can be collected in a replaceable, inexpensive and disposable bottle. Efficiently separates the liquid and gas phases, increasing the sample recovery rate by as much as 98%, and since no solvent is required for sample collection and capture, it is environmentally friendly and economical, and separate separation. It enables high-speed, high-volume, high-purity collection room collection without the need for facilities.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a collection system including a schematic flow path diagram showing a supercritical flow chromatography system and a sample cassette as an embodiment of the present invention.

FIG. 2 is an exploded perspective view showing a sample collection cassette.

FIG. 3 is a plan view showing the upper surface and the lower surface of the cassette lid.

FIG. 4 is a plan view showing another representative embodiment of an automated partial collection system.

FIG. 5 is a side view showing another typical embodiment of an automated partial collection system.

FIG. 6 is an exploded perspective view showing a reciprocating sample collection cassette, lid and mechanized motion control system.

FIG. 7 is a detailed side view showing a cassette that reciprocates and a mechanical control device.

FIG. 8 (A) is a detailed cross-sectional view showing a state before inserting a transfer tube into a glass test tube. (B) It is a detailed cross-sectional view which shows the state after inserting a transfer tube into a glass test tube.

FIG. 9 shows another embodiment as an integrated collection cassette with multiple rows of collection chambers.

FIG. 10 shows another additional embodiment as a reciprocating collection cassette for an automated system.