JP2003526087A - Multi-channel high-throughput purification apparatus and method - Google Patents

Abstract

(57) Abstract: A multi-channel purification system for purifying multiple samples from a chemical library (10). The system (10) uses chromatography, preferably supercritical fluid chromatography. Four parallel channels are coupled to a common analyzer (16), such as a mass analyzer, and a computer (18). The sample in each channel includes a supercritical fluid chromatography column (32) to separate the sample, an ultraviolet detector (34) to detect sample peaks inside the sample stream, and an analyzer It passes through a micro sample valve (38) diverting to (16) and a fraction collection valve (43). The remainder of the sample goes to the fraction collection valve (43), while the mass analyzer (16) analyzes the diversion to determine if the target compound is inside the sample portion. Depending on whether the target compound is present, the fraction collection valve (43) sends the sample portion to one of the two fraction collectors (22, 24).

Description

Detailed Description of the Invention

TECHNICAL FIELD The present invention relates to sample purification devices and methods, and more particularly to high throughput purification devices and methods for samples from chemical libraries.

BACKGROUND OF THE INVENTION The relationship between molecular structure and function is a fundamental part of the study of biology and other chemical systems. Structure-function relationships are important for understanding, for example, enzyme functions, cell transmission, cell regulation, and feedback mechanisms. It is known that certain macromolecules interact and bind with other molecules that have a specific three-dimensional space and electron distribution. Macromolecules with such specificity are
A protein, glycoprotein or antibody, or an oligonucleotide sequence of DNA or RNA, etc., can be regarded as a receptor. Various molecules that bind to receptors are known as ligands.

A common method of producing ligands is to synthesize molecules in a liquid phase or on a solid phase resin in a stepwise fashion. Since the introduction of liquid-phase or solid-phase synthesis methods for peptides, oligonucleotides and small organic molecules, new methods have been developed using liquid-phase or solid-phase technology, using automation or technique techniques, thousands of species, In some cases millions of individual compounds can be produced. A collection of compounds is commonly referred to as a chemical library. In the pharmaceutical industry, chemical libraries of compounds are formatted in 96-well microtiter plates. This 96-well format has become standard in nature and provides a convenient way to screen these compounds to identify novel ligands for biological receptors.

In the recently developed synthetic method, it is possible to construct a large chemical library in a relatively short time as compared with the conventional synthetic method. For example, automated synthetic methods in sample production have allowed the production of up to 4,000 compounds per week. However, samples containing these compounds were not
% To 60% of impurities are typically included. When samples containing these impurities are screened against selected targets such as novel ligands or biological receptors, these impurities can give erroneous screening results. As a result, samples that give a positive result in the first screen must be further analyzed and screened to verify the accuracy of the first screening result. This verification process requires that additional samples be available. The verification process also increases the cost and time required to accurately verify the presence of the target compound.

Samples can be purified to achieve purities of 85% or higher. Screening of purified samples provides more accurate and meaningful biological results. However, conventional purification methods are extremely slow and expensive. For example, conventional purification methods using high pressure liquid chromatography (HPLC) require approximately 30 minutes to purify each sample. Therefore, purification of 4,000 samples produced in a week requires at least 2000 hours (ie 83.3 days or 2.77 months).

[0006] Furthermore, a conventional purification method such as HPLC requires a large amount of solvent and generates a large amount of waste solvent. The disposal of solvents, especially halogenated solvents, must be carefully controlled for legal and environmental reasons, and thus the disposal process can be laborious and extremely costly. Disposal of non-halogenated solvent is somewhat less severe. Therefore, when using a halogenated solvent and a non-halogenated solvent, the waste solvent is separated. However, separation of large volumes of solvent can be a difficult method to perform efficiently and inexpensively. Therefore, purification of large chemical libraries can be economically prohibitive. Therefore, there is a need for a faster and more economical method of purifying each sample of a large chemical library.

Supercritical fluid chromatography (SFC) offers a faster purification method than HPLC. SFC uses a multiphase fluid stream containing a gas such as carbon dioxide in the supercritical state, a carrier solvent and a selected sample. The fluid stream is analyzed through a chromatographic column, then in a bulk sought probe for the target compound. SFC
The solvent and sample are loaded onto the gas, which is advantageous because the amount of solvent required during the purification procedure is substantially less than the solvent volume used in the HPLC. In addition, the amount of waste solvent at the end of the operation is substantially small, and therefore, the waste solvent to be treated can be small. However, SFCs require pressure and temperature regulation that are difficult to control accurately and consistently over time. There are many different types of refiners. These devices typically have commonality in that the sample is sent to a chromatography device that separates the compounds in a timely manner, and the fraction collector collects the target compound. These devices must be able to process large numbers of samples, and also large numbers of substances and solvents, in order to maintain high throughput processes. Traditionally, semi-preliminary or preparative scale chromatography systems have been specialized for typical milligram synthesis. While this is achievable, it is less feasible under high throughput conditions because several problems have become apparent in its practice: use of large amounts of solvent, Large amount of waste solvent generated, large costly perforated column, and relatively large collection capacity of target compounds. Sufficient chromatographic purification will not be achieved without the proper flow rate or column size.

A further disadvantage experienced in high throughput purification techniques is the durability of equipment components that regulate the high pressure, volume or flow rate of the sample through the purification equipment. The purifier is
Extreme accuracy and extremely high tolerance are required to ensure cross-contamination and to obtain purified compounds. Thus, each component of the device must be sufficiently durable to accept the aggressive environment while producing the exact results required. If each component of the device is not durable enough and needs to be destroyed or repaired too soon, the component must be shut down for replacement or repair. A further drawback experienced in the routine purification process of large chemical libraries is sample management during the purification process. For example, chemical libraries are typically maintained as multiple sets of 96-well microtiter plates, each well containing an individual sample. Each sample is tracked by the "well address" of the microtiter plate. When removing a sample or a portion of a sample from selected wells of a microtiter plate for purification, the purified sample is collected and processed in a separate container and finally used for receiving similar microtiter plates. Returned to the well. The receiving well is preferably
Corresponding well addresses in the microtiter plate so that the accuracy of the library record for the sample position in each microtiter plate can be maintained.

A conventional purification method is that the large collection volume of fluid (eg, solvent containing purified sample) is greater than the volume of the receiving wells of a typical microtiter plate,
Reformatting of purified samples is typically required. The large collection volume should be reduced to fit the wells of the receiving microtiter plate. The depleted purified sample-containing fluid is also tracked and placed in the appropriate wells of the receiving microtiter plate that accurately maps the sample to the well locations taken at the beginning of the purification procedure. Reformatting such purified samples into receiving microtiter plates increases the time and cost of the purification method. Therefore, there is a need for a purification method that enables rapid and economical sample purification by collecting the purified sample directly on a microtiter plate that is directly mapped to the original microtiter plate.

DISCLOSURE OF THE INVENTION The present invention relates to a multichannel high throughput purification apparatus and method for samples from chemical libraries that overcomes the deficiencies experienced in the prior art. In one exemplary embodiment of the invention, the multi-channel high throughput purification method of the invention simultaneously purifies multiple, eg, four, samples from a chemical library.

The method of the above exemplary embodiment simultaneously purifies all four samples by supercritical fluid chromatography (SFC) in four channels of the purifier.
The method is to pass a first sample along the SFC channel of the first channel, separate the first sample into sample subdivisions, each sample subdivision along at least a portion of the first fluid flow path. And isolating each other. The supercritical fluid pressure in the fluid flow is
Adjust with back pressure regulator and pressure relief valve. Also, this method
Moving the separated sample aliquots along the fluid flow path and detecting at least one sample aliquot flowing along the fluid flow path. In addition, the method redirects the sample from the sample aliquot, directs the sample to the analyzer while the rest of the sample aliquot is flowing along the fluid flow path, and directs the sample to the analyzer. Analyzing and determining if one sample aliquot has selected sample characteristics. The method may also collect one sample aliquot into a first receiver, such as a well of a first microtiter plate, only if the sample aliquot has a selected sample characteristic. Including. If the sample aliquot does not have the selected sample characteristic, then the sample aliquot is collected in a second receiver, such as a corresponding well in the second microtiter plate.

The exemplary multi-channel high throughput purification method above further comprises purifying the second sample along the second channel concurrently with the purification of the first sample. Purification of the second sample is performed by passing the second sample along the second channel of the second channel, separating the second sample into sample subdivisions, and subdividing each sample subdivision into at least a part of the second fluid flow channel. Along with isolating each other. The method also includes moving the separated sample subdivisions along the second fluid flow path and detecting at least one sample subdivision flowing along the second fluid flow path. The method includes adjusting a second sample pressure along the flow path. In addition, the method involves taking a sample from one sample aliquot and directing this sample to the same analyzer used in the first channel. The rest of the sample subdivision
2 Continues to flow along the flow path.

The method also includes analyzing with an analyzer, wherein each of the first and second samplings is analyzed separately according to a selected analysis order protocol. First
Analysis of the two samples determines whether the sample aliquot has the selected sample characteristics. The method involves collecting the sample aliquot in another receiver, such as another well in the first microtiter plate, only if the sample aliquot has a selected sample characteristic. . If the sample subdivision does not have the selected sample characteristic, the sample subdivision will
Collect in another receiver, such as another well in the second microtiter plate.

In one embodiment of the invention, the high throughput purification method comprises the steps of:
Purifying the third and fourth samples along the corresponding third and fourth channels in a manner similar to the purification described above in connection with the sample of. In this embodiment, the same analyzer is used to analyze samples from all four samples. All samples are analyzed separately according to the selected analysis order protocol.

The present invention also relates to a multi-channel high throughput purification device for purifying multiple samples from a chemical library at substantially the same time. In one exemplary embodiment, the apparatus includes a controller and a sample analyzer coupled to the controller, the analyzer constructed to determine whether each sample has a selected sample characteristic. Has been done. 1st, 2nd, 3rd
And a fourth purification channel is connected to the sample analyzer. The first purification channel includes a separation device that receives the sample stream and separates the first sample into sample aliquots, with each sample aliquot positioned to isolate one another within the sample stream. The detector is positioned to receive the sample stream from the separator and detect at least one sample aliquot within the first sample. An adjustable back pressure regulator receives the fluid flow from the detector and regulates the pressure of the fluid flow in the first channel.

The microsampling device is positioned to receive the sample flow from the back pressure regulator and is moveable between open and closed positions while passing a substantially continuous fluid flow through the device. In the closed position, the microsampling device blocks the fluid flow from passing through the analyzer, allowing the fluid flow to continue to flow into the microsampling device. In the closed position, the microsampling device also passes a substantially continuous stream of carrier fluid through the sampling device to the analyzer. In the open position, the microsampling device directs the sample from at least one sample subdivision to the analyzer for analysis, while the rest of the sample subdivision is substantially uninterrupted through the microsampling device. To move.

A pressure relief valve receives the remaining sample stream from the microsampling device and maintains a selected pressure in the sample stream downstream of the microsampling device. A flow directing valve is connected to the first flow path and is positioned to receive the sample flow downstream of the pressure relief valve. The flow directing valve is moveable to a first position to direct the sample aliquot in one direction if the analyzer determines that the sample aliquot has a selected sample characteristic. .
The flow directing valve can be moved to a second position to point the sample sub-portion in another direction if the analyzer determines that one sample sub-portion does not have the selected sample characteristic. Is. A first receiver, such as a well of a microtiter plate, has one sample aliquot having selected characteristics so that when the flow directing device is in the first position, the sample aliquot from the flow directing device is Position it to receive the section. A second receiver, such as a well in a second microtiter plate, has one sample aliquot that does not have the selected characteristics, so that when the flow directing device is in the second position, Position to receive sample aliquot.

The second purification channel of the purification device includes a separation device positioned to receive the second sample stream and separate the second sample into sample aliquots. Another detector is connected to the separator and positioned to receive the second sample from the separator. The detector is constructed to detect at least one sample aliquot in the sample stream. A microsampling device is positioned to receive the sample flow from the detector and is movable between open and closed positions. The microsampling device, when in the closed position, passes the second sample stream blocking the passage to the analyzer.
In the open position, the microsampling device directs the sample from the sample subdivision toward the analytical analyzer, while the rest of the sample subdivision continues to flow along the second flow path substantially uninterrupted. The back pressure regulator and the pressure relief valve are respectively connected to the second upstream and downstream of the micro sampling device.
Receive a sample stream and selectively control the pressure of the second sample stream along the second purification channel. A flow directing valve is connected to the second flow path and is positioned to receive the sample flow through the second flow path. The flow directing valve is movable toward a first position to direct the sample sub-portion in one direction if the analyzer determines that one sample sub-portion has the selected sample characteristic. . The flow directing valve can be moved to a second position to point the sample sub-portion in another direction if the analyzer determines that one sample sub-portion does not have the selected sample characteristic. Is.

A receiver, such as another well of the first microtiter plate, has a flow directing device when the flow directing device is in the first position because one sample aliquot has selected characteristics. Position to receive the sample aliquot from. Another receiver, like another well of the second microtiter plate,
One sample aliquot does not have the selected characteristic, and is positioned to receive its aliquot sample from the flow directing device when the flow directing device is in the second position.

In one embodiment of the present invention, the purification apparatus of the present invention comprises a third and fourth purification method for purifying the third and fourth samples substantially simultaneously with the first and second samples. Including channels. Each of the third and fourth purification channels is connected to the same analyzer and each sample aliquot is directed to a respective receiver, such as a well of the first and second microtiter plates, as described above.

One aspect of the present invention provides a micro-sample or flow divider valve for use in a high-throughput purifier for purifying selected samples from a chemical library. The purification apparatus of the present invention has a sample channel, a carrier fluid channel, and an analyzer connected to these sample channel and carrier channel.
The micro sample valve includes an internal chamber and a valve body having a sample flow path and a sample inlet and an outlet connected to the internal chamber. The valve body has a carrier fluid inlet connected to the carrier fluid flow path and an external inlet channel connected to the analyzer. The stem is slidably mounted in the inner chamber and is movable between the first and second positions inside the chamber. The stem has a fluid bypass channel that connects the sample inlet and the external inlet when in the sample inlet first position to pass a selected portion of the sample through the analyzer. The stem blocks the carrier inlet when in the first position to prevent fluid from the carrier fluid flow path from migrating to the outer inlet.

When in the second position, the fluid bypass channel in the stem is connected to the carrier inlet and the external inlet to allow the selected carrier fluid to flow from the valve body to the analyzer. The stem, when in the second position, blocks the sample inlet from connecting to the outer inlet to prevent the sample stream from flowing into the outer flow channel.

[0023] One aspect of the invention also includes an automated fraction collection assembly that holds the microtiter plate in a fixed position and dispenses sample components into selected wells in the microtiter plate. The fraction collection assembly includes a dispensing needle that dispenses sample components into a disposable telescoping chamber and then into a microtiter plate. The dispensing needle is mounted on a dispensing head adapted to extend into the disposable telescoping chamber and the sample components are concentrated in the telescoping chamber and then dispensed into the microtiter plate.

The dispensing head is moveable from a pick-up position that picks up the expansion chamber to a collection position on the microtiter plate that dispenses the sample into selected wells of the microtiter plate. The dispensing head is also moveable from the dispensing position to a telescoping chamber dumping position where the telescoping chamber is discharged into the waste container, thereby exposing the dispensing needle. In addition, the dispense head can also be moved to a wash position at the wash station on the fraction collection assembly, where the dispense needle is washed to avoid cross-contamination between samples.

BEST MODE FOR CARRYING OUT THE INVENTION The structure and function of each exemplary embodiment of the present invention can best be understood by referring to the drawings. The same reference numbers are shown in multiple figures. These same reference numbers indicate the same or corresponding structures in each drawing.

A multi-channel high throughput purification apparatus 10 according to one exemplary embodiment is shown in FIGS.
And their respective components are shown in FIGS. Purification device 10 is constructed to simultaneously purify four samples 12 from a chemical library, purifying each sample along each purification channel 14 in the device. Purification in this exemplary embodiment is accomplished by chromatography, and more specifically by supercritical fluid chromatography (SFC), described in more detail below.

Each channel 14 receives a selected sample from the feeding microtiter plate 20. Each channel 14 is connected to a conventional analyzer, such as a mass spectrometer 16, which analyzes selected aliquots of each sample according to a predetermined analysis order protocol. In one embodiment, the analyzer is
Includes multiple compound identification devices. In this exemplary embodiment, each feeding microtiter plate 20 includes a bar code or other selected display or tracking mechanism that provides information specific to the feeding microtiter plate.
The purification device 10 is a specific supply microtiter plate used in each purification operation.
It includes a bar code reader 15 for identifying 20 and the like.

Each component of each channel 14, such as the mass spectrometer 16 and bar code reader 15, is coupled to a computer controller 18, which monitors and controls the operation of each component during the purification operation. Mass spectrometer 16
Is also coupled to computer 17, which provides the user with additional control or monitoring capabilities during the refining operation.

After each sample 16 has been analyzed by the mass spectrometer 16, the substantially purified sample aliquots are placed in corresponding wells of the receiving microtiter plate 22 (FIG. 2) or other selected sample collector. Distribute directly to. The remaining portion of the sample, detected by the detector as a crude product, was transferred to the first microtiter plate 24 (
This is also illustrated in Figure 2). Therefore, four samples 12 are removed from the feeding microtiter plate 20 and purified, each sample being placed directly into the corresponding well positions of the two receiving microtiter plates 22 and 24,
One contains the purified target compound and the other contains the crude. In one embodiment, four samples are sequentially withdrawn from the feeding microtiter plate with the same withdrawal needle assembly. In another embodiment,
The four samples are removed at substantially the same time by a withdrawal assembly having four withdrawal needles.

The receiving microtiter plates 22 and 24 have a bar code or the like thereon, and a bar code reader 25 (FIG. 2) is provided adjacent to each receiving microtiter plate. This second barcode reader 25 is also a computer controller 18 (see FIG. 1).
) To identify and track sample dispensed into selected wells of each microtiter plate. Purified target compounds in microtiter plates 22 and 24 can then be screened by a method selected to locate specific target compounds.

The microtiter plate 22 is rigidly held within an automated fraction collection assembly 23 connected to a computer controller 18 (FIG. 1). The fraction collection assembly 23 condenses the vaporous sample components and then the microtiter plate 22.
Or automated and built to pick up a clean disposable or reusable telescoping chamber that feeds into 24. Fraction collection assembly 23 includes a wash station in which the sample dispense needle is fed to each microtiter plate and the next set of clean telescoping chambers is moved to the next sample dispenser. Clean before picking up for shipment.

In the above exemplary embodiment, selected microtiter plates
20 is specified by the bar code reader 15 and is located on the auto sampler 21 (FIG. 1). In one embodiment, autosampler 21 is a Gilson 215 autosampler manufactured by Gilson, Inc., Middleton, Wisconsin. As best seen in the block diagram of FIG. 3, each sample was removed by the autosampler 21 from selected wells of the feeding microtiter plate 20 into one sample channel 30 of each of the four channels 14. Supplied. 4 samples 12
It is introduced into each purification channel 14 at substantially the same time. The illustrated embodiment is 4
Although individual samples 12 are purified at substantially the same time, other numbers of samples can be simultaneously purified by the apparatus according to the present invention.

As best seen in FIG. 3, sample 12 is mixed with carbon dioxide from a CO ₂ source 29 and modifier solvent from a solvent source 33 and flows into each channel 14 at a selected flow rate. To generate a carrier flow. The carbon dioxide stream from heat exchanger 36 is cooled by recirculating cooling bath 35 and drawn into mixer 39 by CO ₂ pump 37. The CO ₂ flow also passes through the pulse damper to minimize pulsations that could have been produced by pump 37. The modifier solvent flows from solvent pump 41 into mixer 39 where it mixes with carbon dioxide. The carbon dioxide / solvent mixture then flows to the sample injection valve 43, where the sample 12 is received from the autosampler 21 and mixed with the carrier stream to produce the sample stream 31.

The sample stream 31 is passed through a heat exchanger 45 (at which point the fluid is supercritical) and then through a separation medium such as the SFC column 32, which allows each sample in the sample stream 31 to flow. Separate the ingredients. Thus, each sample component separates from the other components as the sample stream exits the SFC column and passes through the purification channel 14.

In one embodiment of the invention, the column 32 has a column, as shown in FIGS.
It is a two-component column used in supercritical fluid chromatography. As best seen in FIG. 4, each component of column 32 includes an upper dilution body 400 that forms a dilution chamber 408. The top of the dilution body 400 is connected to the inlet tube 410 through which the sample stream 31 travels to the column 32. The upper dilution body 400 is connected to the charging body 402 and is held securely by a top end cap 401 that is externally wound onto a thread of the charging body 402. In another embodiment for use in liquid chromatography, no diluting chamber is needed and thus column 32 does not include a diluting body connected to the load.

The dilution chamber body 400, top end cap 401, and load 402 of the above exemplary embodiment
Is manufactured from an inert material such as stainless steel. In another embodiment, other inert materials can be used in the construction of each component of the column. The separation body 403 at the upper end is connected to the lower portion of the charging body 402.
The lower end of the separation body has an outlet tube 412 where the separated sample stream 31 exits the column 32.
Is rigidly connected to the bottom end cap 404 which is connected to.

As best understood in the cross-sectional view of FIG.
From the top threaded inlet 505 to the inlet 505 where the inlet tube 410
Sealed by an outer ferrule seated on the inner top ferrule sealing point 506. The sample stream is directed radially from the inlet tube 410 into the upper dilution chamber 408 by the inverted top funnel 507. The top funnel portion 507 is substantially conical in shape and forms the top of the dilution chamber 408. The body of dilution chamber 408 is substantially cylindrical, but may be constructed with other geometric shapes in alternative embodiments. Dilution room 40
The bottom of 8 has an inverted funnel portion 509 that projects radially outward from the dilution chamber body.
Thus, the bottom funnel 509 overhangs the lower opening, which has a larger diameter than the dilution chamber body. The bottom opening of the bottom funnel 509 has a top frit 51 below the dilution chamber 408.
Positioned above 0.

The total volume of the dilution chamber is free of stationary phase material. Dilution of the sample in the sample stream occurs as the sample stream travels downwardly from the body to the bottom funnel 509, from where it passes to the top frit 510. The top frit 510 distributes the sample on the column bed 512 in the loading area 520 just below the top frit 510. The sealing of the diluting chamber 408 is achieved at the top frit 510 where the diluting chamber body 400 internally fits into the charging body 402.

The loading body 402 has a loading area 520 below the top frit 510 and a transition area 522 below this loading area. The charging region 520 and the transition region 522 in the charging body are filled with a stationary phase material such as cyano forming a column bed 512 within the column 32. In other embodiments, other stationary phase materials may be used to form the column bed 512. The charging region 520 has an inner diameter that is approximately twice or more the inner diameter of the separation region 524 and a length that is approximately half the length of the separation region or less. In the charging zone 520, the sample stream traverses downwards from the column bed 512 towards the transition zone 522 having the conical shape formed by the charging body 402. The transition region 522 directs the sample stream into the separation region of the column bed 512.

The top of the separation body 403 is screwed and joined to the bottom of the charging body 402 by a screw joint, and is sealed by a joint frit 511 sandwiched therebetween. Separation body 403 in the illustrated embodiment is made of stainless steel and is formed such that the interior chamber containing separation region 524 of column bed 512 has a slanted cylindrical shape with a wide upper end and a narrow lower end. The inner chamber of the separation area 524 of the column 512 is filled with the stationary phase material. The sample stream travels downwardly from the column bed 524 in the separation region 524 through the bottom frit 513 and onto the bottom fluid funnel 514 formed in the bottom end cap 404. The bottom of the isolation region 524 is sealed by a bottom end cap 404 that is externally screwed onto the isolation body 403. The bottom frit 513 has a bottom end cap
It is sandwiched between 404 and separation body 403. The bottom fluid funnel 514 is conical and directs fluid into a bottom threaded port 516 formed in the bottom end cap 404; the threaded port 516 can be threaded with an outlet tube 412. Outlet tube 412 is outlet 5
If screwed into 16, use an outer ferrule to seal to the bottom end cap 404 at the bottom ferrule closure point 515.

In another embodiment, shown in FIG. 6, column 32 is a “configured” column.
In terms of similarity between the two embodiments, the same components in the two embodiments are shown in the drawings with the same reference numbers for clarity. The one-component column is substantially the same as the two-component column described above, except that the loading body 602 and the separation body 603 are integrally constructed from a single stainless steel unit to provide one-piece loading and separation.
= OPLAS) structure 617 is formed. Therefore, the upper frit 511 used in the two component column is not needed and is omitted.

As best seen in the cross-sectional view of FIG. 7, the dilution chamber body 400 fits internally into the OPLAS structure 617, with a top end cap 401 externally threaded onto the OPLAS body. It is fixed. The lower end of the OPLAS structure 617 is internally threaded into the bottom end cap 404. Therefore, the loading region 520 formed in the OPLAS structure 617 has a diameter that is about twice or more the inner diameter of the separation region 524 and about half the length of the separation region or less. 8A-C graphically show the results of three chromatographic runs showing the improvement over the prior art obtained by column 32 according to the present invention. All three chromatographic operations were carried out under the same chromatographic conditions by injecting the same charge of the three compound mixture. Run 200 (Figure 8A) shows the separation results using a single prior art column injected with a small volume of solvent mixture. Run 201 (FIG. 8B) shows the separation results using the same prior art single column as run 200 and injecting this column with a large volume of solvent mixture. Operation 202 (FIG. 8C) shows the separation results using the two-component column 32 according to the embodiments of the invention described above. Operation 202 was injected with the same large volume solvent mixture as operation 201.

The first portion of column 32 (eg, the loading and transition portion) has a larger inner diameter and shorter length than the second portion of the column (separation region). Thus, the column 32 according to the present invention can process large volume solvent mixtures containing multiple compounds and can provide highly accurate separation and detection of various compounds such as by use of mass spectrometers and the like. This accuracy, coupled with the corresponding speed of processing large volume solvent mixtures containing multiple compounds, provides faster and more efficient throughput.

Referring again to FIG. 3, the sample stream 31 exits the SFC column 32 and another heat exchanger 4
It flows into the detector 34 through 7. Detector 34 is adapted to detect various compounds or peaks in sample stream 31 separated from each other by SFC column 32. In the illustrated embodiment, the detector 34 is an ultraviolet (UV) detector. Although a UV detector is used in the illustrated embodiment, other detectors can be used, such as an infrared (IR) detector or any other suitable detector capable of identifying peaks in the sample stream 31.

Each detector 34 connects to a conventional computer controller 18. When the detector 34 identifies a peak, it signals the computer controller 18 to indicate the peak. Since the sample flow rate is known in each channel, the computer controller 18 controls each channel 14 as the sample flow 31 passes through the channel.
The position of each peak within can be calculated. For example, if two peaks are detected in the same sample stream 31, computer controller 18 calculates and monitors where each peak is in channel 14. Computer controller 18 also calculates where each peak is relative to each other during the overall purification process.

The sample stream 31 is in a vapor state as it moves through the purification channel. After the sample stream 31 exits the detector 34, additional solvent, referred to as make-up solvent 49, is added to the sample stream when necessary to increase the liquid volume in the sample stream and the fractional collection of the sample. Facilitates transportation to collection assembly (discussed below). Makeup solvent 49 is pumped from the solvent container by solvent pump 51 into each purification channel 14. The solvent container and solvent pump 51 are each connected to a computer controller 18 so that the computer controller can monitor the solvent volume used and control the solvent pump as needed in the selected purification operation.
The computer controller 18 can also monitor the amount of make-up solvent 49 required in the purifying channel during operation to detect if a potential problem has occurred and to give an alarm or other warning to the operator of the device. To be able to

After optionally adding make-up solvent 49 to sample stream 31, the sample stream is
Pass through the back pressure regulator module 53 in the back pressure regulator assembly 55. The back pressure regulator module 53 senses and controls the back pressure in the channel 14 to maintain the desired pressure in the channel.

As best seen in FIG. 9, the backpressure regulator assembly 55.
Includes a housing 900 movably holding four back pressure regulator modules 53 for each purification channel. Assembly 55 also includes a connection panel 902 to which back pressure regulator module 53 is coupled for connection with computer controller 18 (FIG. 3). Module 53 is connected to housing 900 again with connection panel 90
2 is plugged on. Thus, if new or replacement modules 53 are needed in the refinery, they can be installed quickly and easily when no one module is plugged and when plugged into the replacement module.

As best seen in FIG. 10, the back pressure regulator module 53 includes a housing 1002 that houses and protects the cooling grater assembly 1004. The regulator assembly 1004 controls the back pressure within the sample stream as the sample stream passes through each purification channel 14. The regulator assembly 1004 is
It is electrically connected to a stepper motor controller 1006 which activates and regulates the regulator assembly as needed during the refining operation. The stepper motor controller 1006 is a printed circuit board that is also connected to the housing 1002.
It is connected to 1008. The printed circuit board 1008 includes a plurality of connectors 1010 releasably plugged into the connection panel 902 (FIG. 9) of the regulator assembly. Therefore, the computer controller 18 uses the reverse pressure regulator module 53.
To the regulator assembly 1004 by the stepper motor controller 1006.

The back pressure regulator module 53 also includes a front plate 1012 attached to the housing 1002. The front plate 1012 has an inlet 1014 in which the tubing of the purification channel extends to allow the sample stream 31 to pass into the back pressure regulator module 53. The sample stream passes through pressure sensor 1013, which is connected to printed circuit board 1008 to identify the pressure of the sample stream. After the sample stream 31 enters the regulator assembly 1004 and corrects the pressure of the sample stream as needed, as described in more detail below, the sample stream flows from the backpressure regulator module 53 to the outlet 1018 on the faceplate 1012. Exit through.

As best seen in FIGS. 11 and 12, regulator assembly 1004 includes stepper motor 1100 with defeat 1102 connected to stepper motor controller 1006 (FIG. 10). The stepper motor 1100 is a motor mount interconnecting this stepper motor to a back pressure regulator 1106.
It is connected to 1104. The back pressure regulator 1106 has multiple mounting screws that extend through the motor mount 1104 and are screwed into the housing of the step motor 1100.
Firmly held by 1108.

The regulator assembly 1004 also heats the sample stream in the tube of the purification channel to prevent the formation of ice crystals or the like that may occur as a result of the pressure differential across the pressure regulator. Also includes. Heater 11
10 includes a heat conductor 1112 extending over the back pressure regulator 1106, and a heater band 1114 clamped onto the heat conductor by a band clamp 1116. The heater band 1114 is connected to the computer controller 18 so that the heater band regulates its temperature to provide various heating conditions to the back pressure regulator during the refining operation. The thermal conductor 1112 includes a temperature sensor 1118 that monitors the temperature of the thermal conductor during the refining operation. The temperature sensor 1118 is connected to the computer controller 18 (FIG. 3) to allow the computer controller to regulate heat from the heater band 1114 when needed during operation of the regulator assembly 1004.

As best seen in FIG. 12, the regulator 1106 is a
It has an inlet 1200 that receives a purification tube 1201 carrying 31. The inlet 1200 has an inlet channel 1202 that connects with a nozzle 1204 below the inlet. Nozzle 1204 in the illustrated embodiment is a ceramic component that has a diamond coating to provide a very hard and durable nozzle in the regulator. Nozzle 1204 is exposed to caustic solvents and pressures of 2000 psi (140.6 kg / cm ² ) and above, which are extremely harsh. The inlet 1200 is threadedly connected to the nozzle holder 1205 such that the inlet 1200 is easily movable to provide acceptance to the nozzle 1204 if nozzle replacement is required.

Nozzle 1204 includes an inlet channel 1211 extending therein, which is in communication with a microchamber that receives sample stream 31 from the channel. The lower end of the inlet channel 1211 forms a nozzle orifice through which the sample flow passes. The stem 1208 below the nozzle 1204 extends through the seal 1210 into the compartment 1206 and terminates immediately below the nozzle orifice at the lower end of the inlet channel 1211. The stem 1208 is moveable with respect to the nozzle orifice and the regulator 12
Adjustably close the flow path through 06. In the illustrated embodiment, the stem 1208
Is a sapphire stem. In another embodiment, the stem 1208 may be made of other ultra-hard materials such as diamond, ruby and the like. The stem 1208 has a nozzle 12
It is movable relative to 04 and adjusts the size of the opening to adjust the pressure of the sample stream 31.

The sample stream 31 travels from the nozzle 1204, through the orifice, and into the outlet channel 1212, which is in communication with the chamber 1206. The outlet channel 1212 receives the outlet tube 1201 therein and outputs the sample flow 31 from the regulator 1106.
Extends through 14. The outlet tube 1201 extends around the outlet 1214 and surrounds the thermal conductor 1112 approximately twice as to heat the outlet tube, thereby preventing the formation of ice crystals within the purification tube and condensation outside the outlet tube. is doing. A purification tube 1201 extends from the heat conductor 1112 to an outlet 1018 on the front plate 1012 (FIG. 10) of the regulator module as described above, away from the regulator assembly.

In the illustrated embodiment, the stem 1208 includes a regulator assembly 10
It is a sapphire stem with hardness characteristics suitable for use in the high pressure and harsh environment of 04. The sapphire stem 1208 is connected at its lower end to a rod movably positioned within a retaining member 1220 having a threaded lower end. Holding member 12
20 is a Belleville (B) that biases rod 1218 and stem 1208 with respect to nozzle 1204.
ellville) Biasing members 1222 such as washers, wave washers, etc.
including. When the stem 1208 is in direct contact with the nozzle 1204 or is exposed to extremely high pressure pulses, the biasing member 1222 causes the operating sapphire stem 1208 to operate.
Alternatively, the nozzle 1204 is contracted so as to avoid deterioration. However, the biasing member
1222 has sufficient spring stiffness that it does not contract at normal sample flow pressure within the tubes of purification channel 14 during the purification operation.

Adjustment of the regulator assembly 1106 is provided by a double coaxial screw that moves the stem 1208 relative to the nozzle 1204. As best seen in FIG. 12, retaining member 1220 is threaded into an internal thread 1230 formed within shaft 1224 of adjusting screw 1226. In the illustrated embodiment, the internal threads 1230 have a 28 thread pitch per inch (2.54 cm), or a pitch of 28 tpi. The adjusting screw shaft 1224 also has external threads 1232 that thread into the threaded holes in the regulator body 1106. In the illustrated embodiment, the external threads 1232 have a pitch of 27 tpi. Therefore, the external threads 1232 of the adjusting screw 1226 have a thread pitch different from the number of pitches of the internal threads 1230. The internal thread 1230 and the external thread 1232 are both oriented in opposite directions to form a double coaxial screw shape for the damped movement of the stem 1208 relative to the nozzle 1204 with each revolution of the adjusting screw. It is a right-handed type pitch screw thread.

The adjusting screw 1226 has an internal drive spline 1234 that tightly interlocks with the drive spline 1236 on the stepper motor 1100. The drive splines 1236 fit within the internal drive splines 1234. When the stepper motor 1100 is activated by the computer controller 18 (not shown), the drive spline 1236 rotates, thereby rotating the adjusting screw 1226. 1 adjusting screw 1226
When rotated, the double coaxial screw geometry counteracts the range of motion of the retaining member 1228, or stem 1208. For example, the stepper motor 1100 has one adjusting screw
When fully rotated, the retaining member 1220 moves only one pitch number due to the pitch difference between the internal threads 1230 and the external threads 1232.

In one embodiment, one revolution of the adjusting screw along the external threads 1232 will move the adjusting screw 1226 and the retaining member 1220 approximately 0.0373 inches (0.9474 cm). However, the internal threads 1230 are approximately 0.03571 inches (0.
9070 cm) movement, producing a net movement of approximately 0.0013 inches (0.0330 cm). Therefore, the double coaxial screw geometry within the regulator 1106 provides extremely precise and fine adjustment of the stem 1208 to the nozzle 1204, which in turn creates a counter pressure regulator assembly 10.
Tightly control the pressure regulation in sample stream 31 as it passes through 04.

The back pressure regulator 1004 is constructed to have a minimal amount of dead and unscavenged volume in the production channel extending through it, thus risking cross-contamination in different sample purification operations. To prevent or minimize. The back pressure regulator assembly is extremely durable to withstand the harsh environment encountered during ultra high pressure refining operations, while providing sufficient safety to avoid back pressure regulator degradation in events such as pressure spikes. It is built by a component.

In one embodiment, the stepper motor includes a drive spline 1236, a rotation stop 1226 that prevents movement of the adjusting screw 1226 after a selected position relative to the regulator. The travel stop 1238 is positioned to prevent the stepper motor from driving the sapphire stem 1208 too far relative to the nozzle 1204, thereby preventing degradation from overdriving the stepper motor and collapsing the sapphire stem against the nozzle. .

The illustrated embodiment of the refiner uses a regulator assembly having a dual coaxial screw geometry controlled by a computer controller 18. In another embodiment, the counter pressure regulator assembly 53 may be a standard single regulator with a selected control mechanism.

As best seen in FIG. 3, the sample flow 31 travels from the back pressure regulator assembly 55 to the micro sample valve 38. The micro sample valve 38 is operatively connected to the computer controller 18 and is activated by the computer controller when the peak of the sample stream 31 moves from the micro sample valve. Upon activation, microvalve 38 diverts the sampled portion of sample stream 31 toward loss analyzer 16 for analysis. Sample flow 31
The remaining portion of is substantially unobstructed and continues to flow along the flow path of each channel 14. When each micro sample valve 38 is activated,
It contains the correct peaked components. The mass spectrometer 16 analyzes the sampled portion to determine whether the peak is the target compound.

As the four sample streams 31 move through their respective channels 14 and detectors 34 simultaneously, peaks from the four channels will occur at different times during sample operation. Therefore, the mass analyzer 16 typically accepts samples from the four channels somewhat over time between samples. However, in some cases, more than one detector 34 may detect peaks in the sample stream simultaneously or at overlapping times during sample operation. The computer controller 18 is programmed with an analytical ranking protocol that controls the activation sequence of the micro sample valve 38 when the peaks in different channels 14 occur simultaneously or at overlapping times.
Therefore, according to the ranking protocol, the sampled peaks are
The timing at which the peaks are redirected is controlled so that each peak can be analyzed separately by the same analyzer. In one embodiment, if the peaks from different channels 14 are detected simultaneously, the computer controller 18 activates the micro sample valve 38 at different times to direct each peak to the mass spectrometer 16 sequentially. To do so. Activation of each micro-sample valve can be controlled by modifying the analysis order protocol of the computer controller to allow for sequential sampling.

As best seen in FIG. 13, four micro sample valves 38 are parts of micro sample valve assembly 1300 having four valve modules 1302. Each valve module 1302 includes a micro sample valve 38 for each purification channel 14. Each valve module 1302 is movably accommodated in the housing 1304 and plugged into each connector connected to the connection panel 1306. Connection
1306 is connected to computer controller 18 (not shown) to enable the computer controller to activate each micro sample valve 38.

As best seen in FIGS. 14A and 14B, each valve module 1302 includes a front plate 1400 and an opposite plate 1402 rigidly associated with the micro sample valve 38. The full face plate 1400 has inlets 1404 and outlets 1406 that receive the purification channel tubes and direct the sample flow into and out of the valve module 38.

The micro sample valve 38 includes a valve body 1408 located between a pair of electromagnetic solenoids 1410. Each solenoid 1410 has a computer controller 18 (not shown).
The activation of the micro sample valve is controlled as will be described later. Each solenoid 1410 is sandwiched between a valve body 1408 and an outer mounting plate 1414, and a mounting screw 1416 secures each outer mounting plate to the valve body.

As best seen in FIGS. 15-17, the valve body 1408 includes a sample inlet
It has a 1502, a sample outlet 1504 (FIG. 15), a solvent inlet 1506, and a fluid outlet 1508.
The solvent inlet 1506 is not axially aligned with the fluid outlet 1508. The fluid outlet 1508 connects the mass analyzer 16 to the mass analyzer 16 and delivers fluid exiting the micro sample valve 38 through the fluid outlet to the mass analyzer 16 (FIG. 3). The micro sample valve 38 has a handle 1510 slidably inserted in the internal material 512 in the valve body 1408. Stem 1
510 extends slidably in the valve body 1408 and is connected to the electromagnetic solenoid 1410 at the opposite end. The solenoid 1410 controls the axial position of the stem within the valve body. Solenoid 1410 connects to computer controller 18 (FIG. 3) to allow the computer controller to control or adjust stem axial position. The upper and lower seals 1514 are located on the valve body 1408 adjacent to the solenoid 1410.
Located within, a central plastic sleeve 1516 extends between the upper and lower seals. Stem 1510 extends through upper and lower seals 1514 and plastic sleeve 1516 such that a fluid tight seal is formed therebetween. In the illustrated embodiment, the stem 1510 is pressed into a plastic sleeve, thereby preventing dead space around the stem.

As best seen in FIGS. 16 and 17, the stem 1510 includes a fluid outlet 15
It has a through hole 1518 connected to 08 and further connected to the mass spectrometer 16. The stem 1510 also has an axial groove 1520 on the outer flow surface of the valve body 1408 that is connected to the fluid outlet 1508. The axial groove 1520 extends upward from the through hole 1518 along the stem surface, and is formed to direct the fluid flow upward from the through hole along the stem surface and the axial groove of the central plastic sleeve 1516. The through-hole 1518 is formed to allow either the carrier solvent flow or the peak sampling from the sample flow to pass through the mass spectrometer 16.

With reference now to FIGS. 3, 15 and 16, the solvent inlet 1506 (FIGS. 15 and 16) is
It is connected to carrier solvent line 1602 which is connected to carrier solvent source 1604 (FIG. 3) and carrier solvent pump 1606. The carrier solvent pump 1606 is also connected to the computer controller 18 to control the flow of carrier solvent to the micro sample valve 38. A substantially continuous stream of carrier solvent is provided to the microsample valve 38 during the purification operation. In the illustrated embodiment, a carrier solvent line
The 1602 is connected to a series of four micro sample valves 38, allowing carrier solvent to flow through all of the micro sample valves to the mass spectrometer. Therefore, the carrier solvent flows through the solvent inlet 1506 to the first micro sample valve 38.
Enter (FIGS. 15 and 16) and exit through fluid outlet 1508 (FIG. 16), carrier solvent line 1602.
Return to and enter the next micro sample valve through its solvent inlet. The flow passes through each micro sample valve 38 and then to the mass analyzer 16.

The micro sample valve 38 in each purification channel 14 also has a sample flow 31 therethrough.
Has a continuous flow of. Sample stream 31 enters micro sample valve 38 through sample inlet 1502 (FIGS. 15 and 16) and exits sample outlet 1504 through sample line 1522 extending through valve body 1408 in the immediate vicinity of stem 1510. . Thus, the sample stream 31 in the illustrated embodiment is transverse to the carrier solvent stream.

When the micro sample valve 38 is in the lowered normal position, as shown in FIG. 16, the through hole 1518 is below the sample stream 31 and is disconnected from the sample stream 31. Stem 1510 blocks sample stream 31 from passing from fluid outlet 1508 to mass analyzer 16 (FIG. 3). When the stem 1510 is in the lowered position, a continuous flow of carrier solvent enters the valve body 1408 through the solvent inlet 1506, through the through hole 1518, and the axial groove 1520.
And exits the valve body towards the mass spectrometer 16.

During normal use, when no peak is identified, the micro sample valve 38 remains in this lowered normal position, so only carrier solvent flows through the micro sample valve valve to the mass analyzer 16. When detector 34 (FIG. 3) detects a peak in the sample stream and computer controller 18 activates micro sample valve 38, solenoid 1410 raises stem 1510 from its lowered position, as shown in FIG. The axis is immediately moved to the sampling position. In the raised sampling position, the through hole 1518 in the stem 1510 is connected to the sample line 1522,
Through this sample line, the sample stream 31 is fed into the sample inlet 1502 and the sample outlet.
Move between 1504. Therefore, the flow of the carrier solvent is temporarily blocked, and a small sampling portion of the peak moving in the sample line 1522 is diverted from the sample line to pass from the through hole 1518 to the flow outlet 1508 to block the carrier solvent. Enter the carrier line at the entry position. The sampled portion then flows into the mass spectrometer 16 (FIG. 3) for analysis.

As the peak moves from through-hole 1518 for a selected time as determined by computer controller 18, stem 15210 switches back to the down position (FIG. 16). The solenoid 1410 is activated, thereby immediately moving the stem 1510 to the axially lowered position, so that only the portion of the sample stream 31 received by the mass analyzer 16 for analysis is the peak sampling. When the stem 1510 is returned to the lowered position, the flow of carrier solvent to the mass spectrometer 16 is restarted. Therefore, the mass spectrometer 16 receives a continuous flow of fluid, and the sampled portion is a minute sample valve 38.
When activated, it is efficiently inserted as a segment of its continuous flow.

Axial movement of the stem between the lowered position and the raised sampling position allows for extremely rapid switching between positions, thereby enabling small but very accurate sampling of selected portions of the sample stream. . In the illustrated embodiment,
The minute sample valve 38 is formed so as to switch from the normally lowered position to the raised sampling position and further to the normally lowered position in a time of about 15 to 100 milliseconds. In one embodiment, the time is less than 20 milliseconds, allowing a small sample volume of approximately 2 picoliters or less to be directed at the mass spectrometer 16. In another embodiment, the micro sample valve 38 is configured to be movable from the normally lowered position to the raised sampling position and further to the normally lowered position in less than 1 second. This extremely rapid changeover also minimizes the chance of cross-contamination within the body of the sample stream during sampling intervals of multiple peaks.

The micro sample valve 38 is such that the flow path through the valve body 1408 and the stem 1510 provide substantially no dead space and unscavenged volume that can result in cross-contamination between different samples through the micro sample valve. Designed and built. Therefore, the micro sample valve 38 gives extremely accurate results during the purification process. The micro sample valve 38 is also constructed to rapidly take a small sample portion from the sample flow, thereby minimizing the pressure drop in the sample flow across the micro sample valve 38, in the illustrated embodiment, the micro sample valve 38. Approximately 50 pressure drop across sample valve
less than psi (3.515 kg / cm ² ).

As best seen in FIG. 3, the sample flow 31 in each channel 14 travels from the micro sample valve 38 to a pressure relief valve assembly 41 that controls the flow pressure downstream of the micro sample valve. In the illustrated embodiment, the pressure relief valve assembly 41 has the same structure as the back pressure regulator assembly 55 described above, but without the heater on the back pressure regulator assembly.
In another embodiment, a heater may be used if required as a result of ice formation or a large pressure drop encountered in the device. In yet another embodiment, other back pressure regulators can be used as long as they are sufficiently durable and provide sufficient pressure control at the production valve.

The use of pressure relief valve 41 makes the fluid volume to the analyzer very small, either due to the use of stoma capillaries in the analyzer or the active back pressure regulator. Therefore, the pressure differential is reduced and the flow capacity to the mass analyzer 16 is also reduced.

The sample stream 31 exits the pressure relief valve assembly 41 and enters a flow directing valve referred to as the fraction collection valve 40. Each fraction collection valve 40 has one inlet 42
It has two outlets 44 and 46 and a waste outlet 47. Each fraction collection valve 40 is also operably connected to the computer controller 18. When the portion of the sample stream 31 containing the peaks, as identified by the computer controller, enters the fraction collection valve 40 from the inlet 42, the computer controller directs the fraction collection valve to the first peak in the sample stream. It is activated to control whether it is directed from the outlet 44 or the second outlet 46.

If the mass spectrometer 16 determines that the peak is the target compound, the computer controller 18 activates the fraction collection valve 40 and the fraction collection valve is turned on.
Move to position 1. At this position, the sample portion containing the peak exits the fraction collection valve 40 through the first outlet valve 44. The sample portion is directed toward the fraction collection assembly 43 and collected directly in place in selected wells of the first receiving microtiter plate 22.

If the portion of the sample stream containing the peak passes through fraction collection valve 40 and the peak is crude rather than the target compound, the fraction collection valve is switched to the second position. To direct a portion of the sample stream to the second outlet 46. This sample flow
Portion 31 exits the second outlet 46, passes through the fraction collection assembly 43, and is collected directly into the selected well of the second receiving microtiter plate 24. Sample flow
If part 31 passes through the fraction collection valve and the part does not contain any peaks, the sample stream is sent through waste 47 to waste container 52.

The generator 10 of the illustrated embodiment may automatically dispense a purified sample into selected wells of the receiving microtiter plate 22 or 24, with each sample removing that sample first. Are dispensed into the wells of the receiving microtiter plate having the same relative position as the feeding microtiter plate that initiated Therefore, the purified target compound is placed directly into the well having a well address that corresponds one-to-one with the original sample well. Similarly, the crude is also placed directly into the well of the second microtiter plate with the corresponding well address, thus the crude is collected separately from the purified target compound. Direct collection of this target compound into selected microtiter plate wells avoids further processing and formatting of the purified target compound before placing it in the microtiter plate. Therefore, the efficiency of the purification method is increased and the time and cost requirements are reduced.

Purification apparatus 10 of this illustrated embodiment provides for collection of purified compounds having a purity of 85% or greater. Of course, it is preferable to obtain a sample with a purity as close to 100% as possible. Upon collection of the purified target compounds in the receiving microtiter plate 22, these compounds are ready for use in screening processes or other selected processes.

As best seen in FIG. 20, the fraction collection assembly 43 includes a frame 2000 that supports a docking station 2002 that movably receives receiving microtiter plates 22 and 24. The fraction collection assembly 43 also includes a dispensing head 2004 that moves laterally along a rail 2006 attached to the frame 2000 between several operating positions as described below.

The fraction collection assembly 43 includes a hopper 20 for containing a clean disposable expansion chamber 2010.
Including 08. Fraction collection assembly 43 is constructed to provide expansion chamber 2010 from hopper 2008 to pickup station 2012. The pick-up station 2012 holds the telescopic chamber 2010 in a substantially vertical direction with the open top end 2020 of the telescopic chamber facing upwards. The dispensing head 2004 is movable to a position on the pick-up station, and further, the dispensing needle 2014 on the dispensing head 2004 is movable downward so as to extend into the telescoping chamber. The dispensing head 2004 then captures the expansion chamber 2010 and lifts the expansion chamber 2010 from the pickup station.

As best seen in FIG. 21, dispensing head 2004 moves telescoping chamber 2010 from pick-up station 2012 to a dispensing position on selected wells 2024 in microtiter plates 22 and 24. The dispensing head 2004 is connected to a computer controller 18 which controls the position of the expansion chamber 2010 on the well 2024 to correspond to the well position where the sample was first taken. Dispensing head 2004 includes at least part of expansion chamber 2010 selected well 20
Move down to extend into 24. At the same time lowering the expansion chamber 2010, the sample portion containing either the target or the crude material is dispensed from the dispensing needle 2014 to the expansion chamber 2010
And further into selected wells 2024 in microtiter plates 22 or 24.

As best seen in FIG. 18, dispensing head 2004 of the illustrated embodiment.
Holds two telescopic chambers 2010 in a tubular retaining member 2011 in a releasable manner. The air gripping assembly 2015 includes tubular holding members at positions where the expansion chamber 2010 can be abandoned.
It is linked to 2011. Gripping assembly 2015 linked to pneumatic cylinder 2019
Includes a pair of grippers 2017. The air cylinder 2019 moves the gripper 2017 with respect to the tubular holding member 2011 between a holding position and an abandoned position. In the holding position, each gripper 2017 presses the telescopic chamber 2010 against the tubular holding member 2011 to frictionally hold the telescopic chamber within the tubular holding member. In the abandoned position, each gripper 2017 is positioned to freely move its respective expansion chamber 2010 into and out of the tubular retaining member 2011.

Telescopic chamber 2010 is a tubular member having an open top end 2020 and a tapered open bottom end 2022 releasably interlocked by gripping assembly 2015 of dispensing head 2004. The open bottom end 2022 may be partially located within a selected well of the microtiter plate 22 or 24. The open top end 2020 of the telescoping chamber is positioned so that the dispensing needle 2014 extends into the interior region 2028 of the telescopic chamber. Dispensing needle 2014 is located near the side wall of the expansion chamber, so the distribution needle is not co-located with the expansion chamber. Dispensing needle
The end 2013 of 2014 is angled to point towards the side wall of each expansion chamber.

When the sample component is dispensed from the dispensing needle into the interior region 2028 of the expansion chamber 2010, the sample component is in a nebulized state. The nebulized sample component is the squared end of the dispensing needle.
Through 2013 enters the expansion chamber 2010, the end 2013 directs the sample flow towards the side wall of the expansion chamber. The nebulized sample component condenses as a liquid on the sidewall of the expansion chamber, causing the condensate to move in a downward swirling direction along the sidewall.

Liquid sample components that have condensed and are not nebulized flow from the bottom end 2022 of the open telescopic chamber into selected wells of the microtiter plate 22 or 24. When the nebulized sample components are distributed to the expansion chamber 2010, CO ₂ vapor exits the expansion chamber through its modified top end. In the illustrated embodiment, the expansion chamber is vacuum aspirated to draw CO ₂ vapor from the open top end 2020 of the expansion chamber, thereby avoiding cross-contamination between the channels.

When the sp component is condensed in the stretch chamber 2010, some liquid sample components may remain at the bottom of the stretch chamber due to capillary action at the narrow open bottom end 2022. At this point, the fraction collection valve sends the selected solvent into the expansion chamber to wash away any residual sample components and carry the residual sample into the microtiter plate 22 or 24. After adequately dispensing the sample components, the dispensing head can deliver a blow of low pressure air into the telescoping chamber 2010. This air removes the residual liquid sample from the expansion chamber 2010 to the well 20
Extrude into 24.

As best seen in FIG. 21, after dispensing the sample into the microtiter plate 22 or 24, the dispensing head 2004 moves to the chamber drop position and positions the telescoping chamber rearward of the end of the frame 2000. I will let you. The gripping assembly 2015 of the dispensing head 2004 moves to the abandoned position and the telescoping chamber 2010 falls into a suitable waste container. In one embodiment, the expansion chamber 2010 is discarded. In another embodiment, the expansion chamber 2010 is recycled for reuse.

After the dispensing head 2004 has dropped the telescoping chamber, the dispensing head moves to the needle wash position, as shown in FIG. At this needle wash position, the dispensing head 2004
Located on the paired cleaning station 2030. As can be seen in FIGS. 9-21, the wash station 2030 includes wash tubes 20 for dispensing wash solvent or other solutions, respectively.
Including 31. Wash tube 2031 is dispense head 2004 dispense needle 2014 wash tube 20
Form and position it so that it drops into 31. Then, the washing station 1230 is activated to send the washing liquid to the outside of the dispensing needle 2014. The dispensing head 2004 then stands up and cleans the dispensing needle 2014 when pulled up from the washing tube 2031 from top to bottom. The dispensing head 2004 then returns to the expansion chamber pickup position, as shown in FIG. 20, where it picks up a new expansion chamber and prepares the other sample components for distribution in the microtiter plates 22 and 24.

(Industrial Applicability) The high-throughput purification apparatus 10 of the illustrated embodiment enables relatively rapid purification as compared with a conventional purification method. Purification procedures for selected samples can be performed in approximately 6-8 minutes or faster. Therefore, purification of samples contained in 96-well microtiter plates would be approximately 144-192 minutes. Purification of 4,000 samples produced in one week using the sample production method described above would only require a range of 250-330.3 hours. In contrast, it took 2,000 hours to purify 4,000 samples using the usual purification method. Therefore, the groove throughput refining apparatus according to the present invention allows a significant increase in refining rate. The apparatus is also capable of collecting purified samples directly into the wells of a microtiter plate having a location address corresponding to that of the well in the microtiter plate from which the sample was originally removed. That is, the purified sample is ready for screening or other processing. The result is a significant increase in purification capacity that provides a less expensive purification method.

While particular embodiments of the present invention have been described for purposes of illustration, it can be appreciated from the above description that various modifications can be readily made without departing from the spirit and scope of the invention. Will. Accordingly, the invention is not limited except by the claims.

[Brief description of drawings]

FIG. 1 is a block diagram of a part of a multi-channel high throughput purification device according to one embodiment of the present invention.

[Fig. 2]   FIG. 3 is a configuration diagram of another part of the multi-channel high throughput refiner of FIG. 1.

FIG. 3 is a block diagram of the multi-channel high throughput purification apparatus of FIGS. 1 and 2 having four channels.

FIG. 4 shows an enlarged side view of a two-component column of the purifier of FIG. 3 according to one embodiment of the present invention.

[Figure 5]   Figure 5 shows a cross-sectional view of the bi-component column substantially along line 5-5 in Figure 4.

[Figure 6]   FIG. 6 shows a side enlarged view of a component column according to another embodiment of the present invention.

[Figure 7]   7 is a cross-sectional view of one constituent column substantially along line 7-7 of FIG.

[FIGS. 8A to 8C]   3 shows three chromatographic runs showing an improvement over the prior art.

9 is an enlarged exploded isometric view of the back pressure regulator from the purifier of FIG. 3. FIG.

10 is an enlarged exploded isometric view of the back pressure regulator module from the assembly of FIG. 9. FIG.

FIG. 11 is an enlarged isometric view of the regulator / motor assembly of the back pressure regulator module of FIG.

12 is an enlarged cross-sectional view of the regulator assembly taken substantially along line 12-12 of FIG.

FIG. 13 is an enlarged isometric view of a micro sample valve assembly from the purifier of FIG.

FIG. 14A   14 is an isometric view of a micro sample valve from the assembly of FIG. 13. FIG.

14B is an enlarged exploded isometric view of the micro-sample valve from the assembly of FIG. 3. FIG.

FIG. 15   FIG. 15 is a plan view of a valve body of the minute sample valve of FIG.

16 is substantially line 16 of FIG. 14 with the micro sample valve shown in the unsampling position.
FIG. 16 is an enlarged cross-sectional view taken along line -16.

FIG. 17 is substantially line 17-1 of FIG. 14 with the micro sample valve shown in the sampling position.
FIG. 7 is an enlarged sectional view taken along line 7.

FIG. 18 is an enlarged cross-sectional view of the dispensing head and telescoping chamber from the purifier of FIG. 3 with the dispensing head shown in the dispensing position.

FIG. 19 is an isometric view of the automated fraction collection assembly of the purifier of FIG. 3, shown in the chamber pickup position.

FIG. 20   Figure 20 is an isometric view of the fraction collection assembly of Figure 19 shown in a collection position.

FIG. 21   FIG. 20 is an isometric view of the fraction collection assembly of FIG. 19 shown in the dump position.

FIG. 22   Figure 20 is an isometric view of the fraction collection assembly of Figure 19 shown in a wash position.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) G01N 30/72 G01N 30/72 C 30/74 30/74 E 37/00 101 37/00 101 ZCC ZCC ( 81) Designated countries EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE), AU, CA, JP , US (72) Inventor Lipca William Sea United States California 92131 San Diego Red Rock Drive 10819 (72) Inventor Cracovar Jonathan Dee USA California 92083 Vista Wellington Lane 1926 Apartment 151 F Term (Reference) 2G059 AA01 BB01 BB04 CC16 EE01 HH01 HH03 KK01 MM04 4D017 AA03 AA13 BA03 CB01 DA03 EB01 EB03

Claims (85)

[Claims] 1. A substantially continuous multi-channel high throughput supercritical fluid chromatography purification of at least a first and a second sample from a chemical library to provide corresponding array-to-array mapping of cleaning components. In the method, the first sample is chromatographically separated into a first and a second sample portion, the first and second sample portions are separated from each other along at least a portion of the first fluid passage, and the first and second sample portions are separated from each other. The sample portion is moved along the first fluid passage, the first sample portion is detected along the first fluid passage, the first sampling is removed from the first sample portion, and the rest of the first sample portion is the first portion. The first sampling is sent to the analyzer while staying along the fluid passage, the first sampling is analyzed by the analyzer, and the first sampling portion to the first sampling portion have the first selected sample characteristic. If the first sample part has the selected sample characteristic, the first sample part is collected in the first collector, and if the first sample part does not have the selected sample characteristic, the first sample part is collected. Purifying the first sample along the first channel by collecting in the second collector and chromatographically separating the second sample into third and fourth sample sections, along at least a portion of the second fluid passageway. Separating the third sample part and the fourth sample part from each other, moving the third and fourth sample parts along the second fluid passage, detecting the third sample part along the second fluid passage, 2 samplings are removed from the 3 rd sample section, the 2 nd sampling is sent to the analyzer with the remainder of the 3 rd sample section remaining along the second fluid path, the 2 nd sampling is analyzed by the analyzer, From the sampling, it is determined whether the third sample part has the second selected sample characteristic, and if the third sample part has the selected sample characteristic, the third sample part is collected in the third collector and the third sample part is selected. A method comprising purifying a second sample along a second channel at substantially the same time as purifying the first sample by collecting the third sample portion in the fourth collector if it does not have sample characteristics. 2. The microtiter plate further comprising: removing the first sample from the first well of the feed microtiter plate, having a position where the first well is relative to the feed microtiter plate, and receiving the first sample portion. Collecting the first sample, including directly in the second well of the second well, the second well having a position relative to the receiving microtiter plate that corresponds to the position of the first well in the feed microtiter plate. The method of claim 1, wherein 3. The method of claim 1, further comprising placing a first sample in the first channel, the first sample containing up to about 25 milligrams of selected sample components in combination with up to about 2 milliliters of loading solvent. Method. 4. The method of claim 1, comprising separating the first sample into first and second sample portions and passing the first sample through a chromatography column. 5. The method of claim 1, comprising separating the first sample into first and second sample sections and passing the first sample through a supercritical fluid chromatography column. 6. The method of claim 1, wherein the detecting comprises detecting the first sample portion with an ultraviolet detector. 7. The method of claim 1, wherein the detecting comprises detecting the first sample with an infrared detector. 8. The method of claim 1, wherein the first and second samplings are separately analyzed according to a selective analysis priority protocol. 9. Removing the first sampling of the first sample portion and actuating the valve if the first sample portion is adjacent to a valve in communication with the first fluid passage, and activating the first sampling valve. The method according to claim 1, characterized in that it is sent from the analyzer to the analyzer. 10. The method of claim 1, wherein the analyzing comprises analyzing the first sampling by a mass spectrometer, and the first retentate remains simultaneously along the first fluid path. 11. The method further comprises passing the first sample in the vapor state through the first fluid passage, condensing the vapor first sample portion in the first expansion chamber, and collecting the first sample portion as a liquid in the first collector. The method of claim 1. 12. A second signal is sent from the detector for detecting the first sample portion along the first fluid passage to the controller, and when the first sample is adjacent to the valve, the controller sends the valve to the second. The method of claim 1, comprising sending a signal, actuating a valve in response to the second signal, and sending a first sampling from the first sample portion to the analyzer. 13. The second sample portion is further detected along the first fluid passage,
When the second sample portion is detected, the detector sends a second signal to the controller,
Sending a second signal from the controller to the valve when the second sample section is adjacent to the valve, actuating the valve in response to the second signal, and sending a second sampling from the first sample section to the analyzer; 13. The method of claim 12, wherein 14. The method of claim 1, further comprising collecting the first and third sample portions in a single collector assembly. 15. The pressure of the first sample in the first channel is detected at a position before the first sampling is removed, and the pressure is adjusted to the selected value when the pressure is not the selected value before the adjustment. The method of claim 1, comprising: 16. The method of claim 15, further comprising adjusting the pressure of the first sample in the first channel at a location after the first sampling has been removed. 17. A method of separating a first sample into first and second sample parts and passing the first sample part through a column having first and second column parts, wherein the first column part is the second The method of claim 1, wherein the method has a diameter greater than the diameter of the column portion and the first column portion has a length less than the length of the second column portion. 18. The method of claim 1 further comprising moving the carrier fluid stream to the analyzer and applying a first sampling to a portion of the carrier fluid stream moving toward the analyzer.
The method described in. 19. A high throughput supercritical fluid chromatographic purification of a third sample from a chemical library, further comprising chromatographic separation of the third sample into a fifth and a sixth sample section, the third fluid passageway comprising: The fifth and sixth sample portions are spaced apart from each other along at least a portion of the fifth fluid passage and substantially simultaneously with the movement of the first and second sample portions along the first fluid passage.
And a sixth sample portion are moved along the third fluid passage, the fifth sample portion is detected along the third fluid passage, the third sampling is removed from the fifth sample portion, and the fifth sample portion is removed. Sending a third sampling to the analyzer with three residues remaining along the third fluid path, analyzing the third sampling by the analyzer, and analyzing the first, second and third samplings according to the selected analysis priority protocol Then, it is determined whether the fifth sample portion has the third selected sample characteristic based on the third sampling, and if the fifth sample portion has the third selected sample characteristic, the fifth sample portion is collected in the fifth collector. , Collecting the fifth sample portion in the sixth collector if the fifth sample portion does not have the third selected sample characteristic, thereby substantially simultaneously cleaning the first and second samples. The method of claim 1, comprising purifying 3 samples along the third channel. 20. The method of claim 19, comprising separating the second sample into third and fourth sample parts and passing the second sample through a chromatography column. 21. The method of claim 19, wherein the detecting comprises detecting the second sample portion with an ultraviolet detector. 22. Removing the third sampling of the fifth sample section and actuating the second valve when the fifth sample section is adjacent to the second valve in communication with the third fluid passage, 20. The method of claim 19, wherein the third sampling is sent to the analyzer from the second valve. 23. The method of claim 19, wherein the analyzing comprises analyzing the third sampling by a mass spectrometer, wherein the third retentate simultaneously remains along the third fluid path. 24. The method further comprising passing the third sample in the vapor state through the third fluid passage, condensing the vapor fifth sample portion in the expansion chamber, and collecting the fifth sample portion as a liquid in the third collector. The method according to 19. 25. A detector for detecting the fifth sample portion along the third fluid passage sends a first signal to the controller, and when the fifth sample portion is adjacent to the second valve, the controller outputs the first signal. 20. The method of claim 19, comprising sending a second signal to the two valves, actuating the valves in response to the second signal, and sending a third sampling from the fifth sample section to the analyzer. 26. Further, the sixth sample portion is detected along the third fluid passage,
When the sixth sample portion is detected, the detector sends a third signal to the controller,
If the sixth sample section is adjacent to the second valve, the controller sends a fourth signal to the second valve, actuates the valve in response to the fourth signal, and the fourth sampling from the sixth sample section to the analyzer. 13. The method of claim 12, comprising sending to. 27. Further, a first signal is sent from the first detector for detecting the first sample portion along the first fluid passage to the controller, and if the first sample is adjacent to the first valve, the controller outputs from the controller. Sending a second signal to the first valve and actuating the first valve in response to the second signal to send a first sampling from the first sample section to the analyzer and a fifth sample section along the third fluid path. The second detector for detecting sends a third signal to the controller, and when the fifth sample section is adjacent to the second valve, the controller sends a fourth signal to the second valve, and in response to the fourth signal, 20. The method of claim 19, comprising actuating two valves and sending a third sampling to the analyzer. 28. The method of claim 27, further comprising passing the carrier fluid stream through an analyzer and applying a first sampling to a portion of the carrier fluid stream moving toward the analyzer. 29. The method of claim 28, wherein the carrier fluid stream passes through the first and second valves. 30. The method of claim 19, further comprising collecting the first, third and fifth sample portions in a single collector assembly. 31. A high throughput supercritical fluid chromatographic purification of third and fourth samples from a chemical library, further comprising separating the third sample into fifth and sixth sample parts, a third fluid The fifth and sixth sample portions are separated from each other along at least a part of the passage, and the movement of the first and second sample portions along the first fluid passage or the third and the third along the second fluid passage. The fifth and sixth sample portions are moved along the third fluid passage substantially at the same time as the movement with the fourth sample portion, the fifth sample portion is detected along the third fluid passage, and the fifth sample portion is detected. Move the third sampling to the analyzer with the third residue remaining along the third fluid path, analyze the third sampling by the analyzer, select first, second and third sampling analysis priority protocol Analyzed according to the third Based on the sampling, it is determined whether the fifth sample portion has the third selected sample characteristic, and if the fifth sample portion has the third selected sample characteristic, the fifth sample portion is collected in the fifth collector, Collecting the fifth sample portion in the sixth collector if the portion does not have the third selected sample characteristic, separating the fourth sample into the seventh and eighth sample portions, and at least a portion of the fourth fluid passage. The seventh and eighth sample parts from each other along the first fluid path and the movement of the first and second sample parts along the first fluid passage or the third and fourth sample parts along the second fluid passage. Moving or moving the fifth and sixth sample portions along the third fluid passage substantially simultaneously with moving the seventh and eighth sample portions along the fourth fluid passage, and changing the seventh sample portion to the fourth. 7th sample detected along the fluid path Moving the fourth sampling of the seventh sample portion to the analyzer with the fourth residue of the sample remaining along the fourth fluid passage, and analyzing the fourth sampling by the analyzer, the first, second, third and The fourth sampling is analyzed separately according to the selected analysis priority protocol, and based on the fourth sampling, it is determined whether the seventh sample section has the fourth selected sample characteristic, and the seventh sample section has the fourth selected sample characteristic. Collecting the seventh sample portion in the seventh collector in the case, and collecting the seventh sample portion in the eighth collector if the seventh sample portion does not have the fourth selected sample characteristic. The method of claim 1, comprising purifying the third sample along the third channel and the fourth sample along the fourth channel substantially simultaneously with purifying the sample. 32. Removing the first sample from a first receptacle in a sample holder, each having a plurality of receptacles in position relative to each other, before the first sample is separated. A first sample in the first fluid passageway, the first collector having a plurality of receptacles each at a position corresponding to the relative position of the receptacles in the sample holder, and collecting the first sample portion. Including collecting a first sample portion in a receiving receptacle in the first collector, the receiving receptacle being at a first position relative to the other receptacles in the first collector corresponding to the first receptacle in the sample holder. The method of claim 1 having in a collector. 33. Removing the first sample from a first receptacle in a sample holder and removing the first sample from one well of a 96-well microtiter plate, the first sample portion being of a first collector. 33. The method of claim 32, wherein collecting in the receiving receptacle comprises collecting the first sample in one well of a 96 well microtiter plate. 34. further comprising removing the first sample from a first receptacle within a sample holder having a plurality of receptacles therein, each of which is in place relative to one another, before the first sample is separated. And a second collector having a plurality of receptacles at respective positions corresponding to the relative positions of the receptacles within the sample holder, and collecting the second sample portion. Including collecting a second sample portion in a receiving receptacle in the second collector, the receiving receptacle having a second position relative to the other receptacles in the second collector corresponding to the first receptacle in the sample holder. The method of claim 1 having in a collector. 35. Further, when the micro sample valve is in the first position, the fluid flow is sent to the analyzer through the micro sample valve, and the micro sample valve is in fluid communication with the first fluid passage and is in the first position. Block the movement of the first sample to the analyzer and also remove the first sampling of the first sample section to move the microsample valve from the first position to the second position, blocking the fluid flow through the valve. 2. The method of claim 1, comprising allowing the first sampling to enter the fluid flow block and moving the first sampling in the fluid flow block to the analyzer. 36. The method of claim 35, wherein the first sampling selector further comprises moving the valve from the first position to the second position after reaching the analyzer through the microsample valve. 37. The method of claim 35, further comprising moving the microsample valve from the first position to the second position and back to the first position within a time period of about 15-100 microseconds. 38. The method of claim 35, further comprising moving the microsample valve from the first position to the second position and back to the first position within less than about 1 second. 39. The method of claim 1 including analyzing the second sampling with an analyzer after the third sample portion is detected after the first sample portion and the first sampling portion is analyzed. 40. Substantially simultaneously with the detection of the first sample portion, the third sample portion is detected, the first and second sampling are analyzed according to an analysis priority protocol, and the second sampling is analyzed by an analyzer. The method of claim 1, wherein the first sampling is analyzed by an analyzer before being performed. 41. In a multi-channel high throughput purification method for a large number of samples from a chemical library, a first sample to be separated into components is moved along a first fluid passage for each different component of the first sample. Passing through a separation medium that will have different transit times, detecting individual components of the first sample as the first sample travels along the first fluid path and identifying the components in relation to the first channel; Sampling the components, sending the sample with its channel identifier to a common analyzer, determining whether the analysis sample has a first selected sample characteristic, and if the detected component of this first sample has a selected sample characteristic, Activating the control valve to allow this first component to collect at the first collector associated with the first channel, If the output component does not have the selected sample characteristic, the control valve is actuated to collect these components in the second collector associated with the first channel and the second sample to be separated into the components in the first fluid passage. The individual components of the first sample as they move along the first fluid path, passing through a separation medium that has different transit times for different components of the first sample. , Identifying the components in relation to the first channel, sampling each component, and sending the sample with its channel identifier to a common analyzer to determine whether the analysis sample has a first selected sample characteristic, If the detected component of one sample has the selected sample characteristic, the control valve is actuated to direct this first component to the first collector associated with the first channel. If the detected components of this first sample do not have the selected sample characteristic, it is possible to collect them and activate the control valve to collect these components in the second collector associated with the first channel, The second sample to be separated is moved along the second fluid passage and is passed through a separation medium whose passage time differs depending on various components of the second sample, and the second sample is moved along the second fluid passage. As it detects the individual components of the second sample, identifies the components in relation to the second channel, samples each component, and sends such a sample along with its channel identifier to a common analyzer where Determining whether or not the detected component of the second sample has the selected sample characteristic, and activating the control valve when the detected component of the second sample has the selected sample characteristic, To collect the second component of the second collector on the first collector associated with the second channel, and if the detected component of this second sample does not have the selected sample characteristic, actuate the control valve to To a second collector associated with the second channel to move a third sample to be separated into components along a third fluid path, the separation medium having different transit times for different components of the third sample. Through, detecting individual components of the third sample as the third sample travels along the third fluid path, identifying the components in relation to the third channel, sampling each component, and It is sent to a common analyzer together with the channel identifier, and it is determined whether the analysis sample has the characteristic of the third selected sample, and the detected component of this third sample is the selected sample If so, the control valve is actuated to allow this third component to collect at the first collector associated with the third channel such that the detected component of this third sample does not have the selected sample characteristic. In some cases, the control valve is actuated to collect these components in the second collector associated with the third channel, move the fourth sample to be separated into components along the fourth fluid passage, and Detecting individual components of the fourth sample as they move along the fourth fluid path through a separation medium that will have different transit times for each component and identify the components in relation to the fourth channel. Then, each component is sampled and such a sample is sent along with its channel identifier to a common analyzer to determine whether the analyzed sample has a fourth selected sample characteristic. If the detected component of this fourth sample has a selected sample characteristic, the control valve is actuated to allow this fourth component to be collected at the first collector associated with the fourth channel. If the detected components of the sample do not have the selected sample characteristic, actuate the control valve and collect these components in the second collector associated with the fourth channel to cause the components of each sample to flow along the first channel. A method consisting of separating and purifying. 42. In a multi-channel high throughput supercritical fluid chromatography purification system for the purification of multiple samples from a chemical library at substantially the same time, each of the samples being separable into a first and a second component. A sample analyzer for analyzing selected sample properties, a first chromatographic sample separation device arranged to separate the first sample stream into first and second sample components, and a first sample in the first sample stream. A first detector arranged to detect the first and second sample components, and movable to send a sampling of the first sample component to the analyzer, the remainder of the first sample portion being in the first flow path. A first diverter along which it flows away from, and a minority of the first or second sample components of the first sample identified by the sample analyzer as having selected sample characteristics. And a second collector arranged to receive a portion and a second collector arranged to receive the residual component of the first sample that does not have selected sample characteristics. A clarification channel, a second chromatographic sample separation device arranged to receive the second sample stream and separate the second sample stream into first and second sample components, and a first sample stream of the second sample stream. A second detector arranged to detect a second sample component, movable to send a sampling of the first or second sample component to the analyzer, the remainder of the second sample part being the second flow path. A second flow diverter along and away from the third flow collector, and a third collector arranged to receive at least a portion of the first or second sample component of the second sample having selected sample characteristics. , The system comprising a second purifying channel coupled to a sample analyzer and a fourth collector arranged to receive the remaining components of the second sample having no selective sample characteristics. 43. A third chromatographic sample separation device arranged to receive a third sample stream and separate the third sample stream into first and second sample components, and a third sample stream of the third sample stream. A third detector arranged to detect the first and second sample components, and movable to send a sampling of the first or second sample components to the analyzer, the remainder of the third sample part being the third A third flow diverter along the flow path therefrom, the first collector being arranged to receive at least a portion of the first or second sample component of the third sample having selected sample characteristics; A third purification channel coupled to the sample analyzer, the second collector being arranged to receive the residual components of the second sample which do not have the selected sample characteristic; and the fourth sample stream. And a fourth chromatographic sample separation device arranged to separate the fourth sample stream into first and second sample components, and to detect the first and second sample components of the fourth sample stream. A fourth detector located at, and a fourth detector portion movable for delivering a sampling of the first or second sample component to the analyzer, and a remainder of the fourth sample portion flowing along the fourth flow path therefrom. A shunt device, the first collector being arranged to receive at least a portion of the first or second sample component of the fourth sample having the selected sample characteristic, and the second collector having the selected sample characteristic. 43. The system of claim 42, comprising a fourth purification channel coupled to a common sample analyzer arranged to receive the residual component of the fourth sample that is not present. 44. The purification system according to claim 42, wherein the sample analyzer is a mass analyzer. 45. The purification system according to claim 42, wherein at least one of the four separation devices is a chromatography column. 46. The purification system according to claim 42, wherein at least one of the four detectors is an ultraviolet detector. 47. At least one of the first and second flow diverters is in fluid communication with the analyzer and is movable between an open position and a closed position to enable sampling of each sample to be sent to the analyzer. 43. The purification system of claim 42, which is a micro sample valve that 48. The expansion chamber of claim 42, wherein the sample is vapor as it passes through the first purification channel, and further the first or second collector receives the vapor sample and condenses it into a liquid prior to receiving the sample. Purification system described. 49. The pressure regulator of claim 42, further comprising a pressure regulator arranged to receive the first sample stream and to regulate the pressure of the first sample stream in the first purification channel upstream of the first diversion device. Purification system. 50. The purification system of claim 49, wherein the pressure regulator has a heater portion that heats at least a portion of the first sample stream. 51. The purification system according to claim 42, further comprising a pressure relief valve position downstream of the first flow divider. 52. Each sample is vapor as it passes through each purification channel and further comprises an expansion chamber for receiving each vapor sample and condensing it into a liquid before the first or second collector receives the sample. The purification system according to claim 42. 53. A sample holder further comprising a plurality of sample receptacles including one sample receptacle, each sample receptacle being in a predetermined position relative to the other sample receptacle, and the collector having a plurality of collecting wells. 43. The method of claim 42, wherein each collection well has a predetermined position with respect to the other collection wells, and the predetermined position of each sample receptacle corresponds to a predetermined position of each of the collection wells. Purification system. 54. A multi-channel high throughput purification system for purifying at least two samples from a chemical library at substantially the same time, and a controller and whether the first sample has selected sample characteristics coupled to the controller. A sample analyzer configured to determine a flow rate, and a separation device arranged to receive a sample stream containing the sample and separate the sample into first and second sample components separated from each other in the first sample. A detector coupled to the separator and arranged to receive the sample stream from the separator and configured to detect at least a sample component in the first sample stream; and to receive the sample stream from the detector. Is positioned and movable between an open position and a closed position, where the open position divides the sampling of sample components. A flow sampling device that directs the remainder of the first sample stream through the flow divider along the flow path, blocking the sample flow from the analyzer in the closed position, and is in fluid communication with the flow path, Is arranged to receive a sample and is movable to a first position to direct the sample portion in a first direction when the analyzer determines that the sample component has a selected sample characteristic, the sample component being the first selected sample At least two purification channels each including a flow directing valve movable to a second position for delivering the sample portion in the second direction if determined by an uncharacterized analyzer; A first collector positioned to receive a sample component having selected sample characteristics from the flow directing device when in the one position, and the flow directing device in the second position If consists of a second collector disposed to receive the sample components that do not have the selected sample characteristics purification system. 55. The purification system of claim 54, wherein the common sample analyzer is a mass spectrometer. 56. The purification system of claim 54, wherein at least one of the two separation devices is a chromatography column. 57. The purification system according to claim 54, wherein at least one of the two separation devices is a supercritical fluid chromatography column. 58. The purification system of claim 54, wherein at least one of the two detectors is a UV detector. 59. The purification system of claim 54, wherein at least one of the two flow sampling devices is a micro sample valve. 60. The sample flow is vapor as it passes through the purification channel and further comprises an expansion chamber for receiving the vapor sample and condensing it into a liquid before the first or second collector receives the sample. The purification system according to claim 54. 61. The purification system of claim 60, wherein the expansion chamber is sized to receive a vapor sample component therein and condense a vapor first sample component along a sidewall of the expansion chamber. 62. The second sample stream is vapor as it passes through the second purification channel, and the vapor second stream is received before the third or fourth collector receives the second sample.
55. An expansion chamber for receiving a sample and condensing it into a liquid.
Purification system described in. 63. A sample holder further comprising a plurality of sample receptacles including a first sample receptacle, each sample receptacle being in a predetermined position with respect to the other sample receptacles, and the first collector being internal to the plurality of collectors. The collector assembly has a predetermined position relative to the other collectors, and the predetermined position of the first sample receptacle in the sample holder corresponds to the predetermined position of the first collector. 55. The method according to claim 54, wherein
Purification system described in. 64. A controller further operatively coupled to the sample analyzer,
55. The purification system of claim 54, comprising four shunt valves and four flow directing valves for controlling sample flow in response to analytical results from the analyzer for components from each channel. 65. In a micro sample valve for use in a high throughput purification system for purifying a sample from a chemical library, the purification system has a plurality of channels, each channel having a sample flow path and a carrier fluid. A sample analyzer in fluid communication with a flow furnace, a sample channel and a carrier fluid channel, a through stem chamber, a sample channel in fluid communication with the sample channel and the stem chamber, a carrier fluid channel and a stem chamber A valve body having a carrier flow channel in fluid communication with the stem chamber and an outlet channel in communication with the analyzer; and a stern slidably disposed within the stem chamber, wherein the stem is between a first position and a second position. Is movable in the sample flow path and allows the selected sample to pass through the valve body without substantially interrupting the sample flow in the sample flow path to the analyzer. Has a fluid bypass in communication with the sample flow path and the outflow channel when the stem is in the first position to allow flow and blocks the carrier fluid flow path when the stem is in the first position for selection A fluid bypass communicates with the carrier fluid flow path and the outflow channel when in the second position to allow the carrier fluid to flow through the valve body to the analyzer, and the sample when the stem is in the second position. A stern engaged with the stem, characterized by blocking the flow channel and having a fluid bypass dimensioned to deliver a small amount of fluid to the analyzer without causing a substantial pressure drop across the microsample valve. , An actuator operable to move the stem between the first position and the second position substantially instantaneously. Npurubarubu. 66. A carrier comprising an inlet and an outlet, and two columns, the first column being connected to the inlet of the carrier tube and having a length less than or equal to 1/2 of the length of the second column. A liquid chromatography system wherein the inner diameter is about twice or more the inner diameter of the second column, and the second column is connected to the outlet of the carrier tube. 67. The system of claim 66, further comprising a dilution chamber located adjacent and in front of the first column, the dilution chamber having a selected geometric interior volume. 68. The system of claim 67, wherein the first and second columns are integrally connected to each other. 69. The system of claim 66, wherein the first and second columns are integrally connected to each other. 70. The system of claim 66, further comprising a Teflon coating on the first and second columns in contact with the fluid. 71. The system of claim 66, further comprising a bonded phase coating covering the portions of the first and second columns that contact the fluid. 72. First and second column parts, wherein the first column part has a first chamber having a first inner diameter and a first length, and the second column part has a second inner diameter and a second length. A chromatography column having a second chamber, the first inner diameter being larger than the second inner diameter and the first length being shorter than the second length. 73. The column of claim 72, wherein the first inner diameter is at least twice the second inner diameter. 74. The column of claim 73, wherein the first length is less than or equal to about half the second length. 75. The column of claim 72, wherein the first length is less than or equal to about half the second length. 76. The column of claim 72, wherein the first and second column sections include a solid phase material. 77. The column according to claim 72, further comprising a dilution chamber adjacent to the first chamber, the first chamber being between the dilution chamber and the second material. 78. The column of claim 72, wherein the first column section has a funnel-shaped transition section connected to the second column section. 79. The column according to claim 72, wherein the first column portion is detachably connected to the second column portion. 80. The column of claim 72, wherein the first column portion is integrally connected to the second column portion. 81. The column of claim 72, wherein the second chamber section is a tapered chamber that tapers inwardly as the second column section moves away from the first column section. 82. In an expansion chamber for collecting a sample in vapor state, allowing expanding gas to escape and collecting condensed liquid, an open tube having a tapered end and means for admitting the vapor into the chamber. An expansion chamber, the means being eccentric within the chamber and facing the proximal tube wall of the open tube. 83. The expansion chamber of claim 82, comprising introducing the vapor at an angle such that a downward spiral of condensed liquid is formed with respect to the chamber. 84. A fraction collector assembly that can be used to collect a portion of a selected sample, comprising a frame and at least one distribution tube movably coupled to the frame and configured to dispense a portion of the selected sample. A dispensing head movable between the pick-up position, the cleaning position and the dispensing position, and a pick-up station formed to detachably hold the selection chamber at a position for engaging the dispensing head when in the pick-up position. And a wash station configured to removably receive the dispensing tube when the dispense head is in the wash position and a receiving container configured to receive a portion of the selected sample when the dispense head is in the dispense position. Fraction collector assembly consisting of docking station. 85. The fraction collector assembly of claim 84, wherein the selection chamber further comprises a storage vessel containing the selection chamber prior to being placed in the pickup station.