The present invention relates to automated sample preparation apparatus for chemical analysis and, more particularly, relates to systems and methods for solid phase extraction for chromatographic analysis.
Sample handling apparatus based on solid phase extraction (SPE) are used for cleanup and enrichment of samples prior to chromatographic analysis. Typical operating procedures of such apparatus includes: 1) eluting a SPE column packed with a sorbent using a solvent to wet the sorbent bed, 2) loading a sample fluid to the column, 3) eluting the column again with a solvent to remove the interfering components from the sample matrices, and 4) washing down the target components in the sample from the column using a solvent and collecting the target components fraction for further chromatographic analysis. The collected fractions may be transferred to a liquid chromatograph (LC) for final analysis manually as in the case of a standalone SPE apparatus or automatically as in the case of a online SPE apparatus.
An online SPE apparatus can transfer the collected fraction to a LC automatically. It can reduce the workload of laboratory staff and also improve the reliability of the analytical results by reducing human errors. Currently available online SPE apparatus may use either one of the two approaches for integration with liquid chromatographic analysis: direct coupling and indirect coupling.
Using the direct approach can achieve high sensitivity since the whole portion of sample loaded to SPE column C2 is used for final LC quantitation. A disadvantage with the direct approach is limited selection of suitable SPE columns. This is because the SPE column needs to have properties similar to the LC column to avoid adverse effects on the chromatographic separation. Also, the SPE column needs to have good lifetime, since it is not easy to change a SPE column that is fixed to a high pressure switching valve and such SPE columns are much more expensive than those used in standalone SPEs. Because of these limits, the direct approach is normally used for samples of simple matrices, such as drinking water.
Some online SPE apparatus based on direct integration approach may use more than one SPE column and switching valve for different sample preparation needs. Examples are presented in U.S. Pat. No. 7,588,725 B2 (Can Ozbal, Donald Green), U.S. application Ser. No. 15/129,543 (Sang-won Lee, Hang yeore LEE), and U.S. Pat. No. 5,468,643A (Syang Y. Su, Gerald K. Shiu). Their common feature is that the SPE columns are fixed on the switching valves and thus have the same limits indicated above for direct integration approach.
In case of the indirect approach, the online SPE apparatus is the same as a standalone SPE apparatus plus a switching valve for injecting the collected fraction to a LC. As demonstrated in
The indirect integration approach can overcome the limits with the direct approach in finding suitable SPE columns. This is because each SPE column is only used for one sample such that the lifetime of the SPE column and the matrices of the sample are not an issue. Also, since only a very small portion of collected fraction from the SPE column is transferred to the LC column, the properties of the SPE column have much less effect on LC separation. Therefore, users have more choices of suitable SPE columns for cleanup and enrichment of their samples. However, the indirect approach does have the disadvantage of lower sensitivity than the direct approach as only a very small portion of a sample is used for quantitation by LC.
Rotary valves are used in sample handling apparatus to divert solvents and sample fluids (e.g., multi-position valves manufactured by VICI Co., Houston, Tex., USA). These rotary valves normally have one inlet port (typically placed in the rotary axis of the valve) and a number of outlet ports that are placed around the inlet port. The rotor inside the valve has a single, radially extending groove that has one end in the rotary centre, thereby connecting the inlet with any one of the outlet ports.
In some applications of sample handling, the flow direction through a SPE column or other components needs to be reversed to accelerate regeneration of the column or to minimize peak broadening of the targeted components during valve switching. The above described common type rotary valves cannot fulfil this task without using additional means, such as a flow redirecting valve or a second pump.
A prior art rotary valve that can reverse the flow direction without using additional means was described in U.S. Pat. No. 8,186,381 B2 (Anders Wilen). This rotary valve can divert fluid to any of the components connected to the valve and can also reverse the flow through a component by rotating the rotor to a certain angle. However, as shown in
Another prior art rotary valve that can reverse the flow direction without using additional means was described in U.S. Pat. No. 8,813,785 B2 (Haibin Wan). The connection ports on this valve are arranged into groups. Each group has at least three ports with one acting as a common port and the others acting as non-common ports. Each port group can be used to connect one SPE column, using the common port as the fluid outlet. This rotary valve can reverse the flow through the SPE columns connected to it by rotating the rotor to a certain angle. As shown in
The object of the present invention is to provide systems and methods for online SPE for LC analysis that can overcome the disadvantages and maintain the advantages of the known online SPE apparatus.
Aspects of the invention include systems and methods that use a multifunctional rotary valve that can divert fluid through the SPE columns in forward and reversed directions.
In some aspects, the system comprises a pump, a multifunctional rotary valve, a 6-port switching valve, and a three way valve. Other components are trays for carrying and moving samples, SPE columns, and the fraction containers, as well as linear actuators for moving the needles and adapters for connecting the SPE columns. With the present 2-stage online SPE apparatus, a sample is first processed using a SPE column as in a standalone SPE. The collected fraction is then loaded onto the second SPE column fixed on a switching valve and directly coupled with a LC instrument. The targeted components from the sample fraction is enriched and further cleaned on the second SPE column before being transferred to LC for final analysis.
The first SPE column is used to remove most interfering materials from the sample matrices. Since it uses conventional columns for standalone SPE, it is much easier to find suitable SPE columns and achieve good cleanup results. The fraction loaded to the second SPE column contains much less interfering materials and thus the lifetime of the second SPE column is extended considerably. Besides, a much larger volume of the collected fraction from the first SPE column is used for LC quantitation thanks to the enrichment effect of the second SPE column and thus achieve good sensitivity.
The core component of the online SPE apparatus is a multi functional rotary valve that can connect the syringe pump with sample, elution solvents, SPE columns, and collected fractions, and can also elute the two SPE columns in two directions. The multi functional valve comprises a stator and a rotor. The stator comprises: a stator interface and a plurality of ports. One of the ports is located in the rotary axis of the valve and designated as central port. The other ports are arranged into a plurality of port groups surrounding the central ports. Each port group comprises at least three ports of which one is designated as a common port and the others are designated as non-common ports. The non-common ports comprise a first non-common port and a second non-common port. Each of the common ports, first non-common ports and the second non-common ports are circularly arranged at the stator interface.
The rotor comprises a rotor interface abutting said stator interface. The rotor also comprises a first channel extending from an axial center of the rotor to at least a point on the rotor interface alignable with the surrounding ports. The rotor also comprises at least two second channels, each second channel extending from at least a point on the rotor interface alignable with a surrounding port to at least a point on the rotor interface alignable with a neighboured surrounding port.
The rotor is coaxially rotatable relative to the stator to configure the fluid selection valve in at least three different connection status; wherein in a first status, the common port of a port group is in fluid communication with the central port via the first channel; wherein in a second status, the common port of a port group is in fluid communication with the first non-common port in the same group via a second channel and the second non-common port is in fluid communication with the central port; and wherein in a third status, the common port of a port group is in fluid communication with the second non-common port in the same group via another second channel and the first non-common port is in fluid communication with the central port.
In one aspect the central port of the fluid selection valve may located in the axial center of the stator. The first channel and the second channels may extend in a plane generally orthogonal to the axis of the rotor. The rotor and the stator may each comprise a substantially disc-shaped body. The first channel and the second channels may comprise generally linear grooves on a surface of the rotor interface, or bores within the rotor below a surface of the rotor interface.
The circular arrangements of the common ports, first non-common ports and the second non-common ports at the stator interface may overlap. The plurality of ports may comprise four port groups, and each port group may comprise three ports of which one is designated as the common port and the other two are designated as the non-common ports, and wherein the rotor comprises one first channel and two second channels, wherein the length of each first channel is equal to a distance of a non-common port to the central port, and wherein the length of each second channel is equal to a distance of the common port to the non-common ports within the same port group.
The common ports may be located on a first imaginary circle, the non-common ports may be located on a second imaginary circle, and the first circle may be concentric but non-overlapping with the second circle.
In drawings which show non-limiting embodiments of the invention:
Throughout the following description, specific details are set forth in order to provide a more thorough understanding of the invention. However, the invention may be practiced without these particulars. In other instances, well known elements have not been shown or described in detail to avoid unnecessarily obscuring the invention. Accordingly, the specification and drawings are to be regarded in an illustrative, rather than a restrictive, sense.
According to aspects of the invention, the two-stage online SPE apparatus comprises a syringe pump, a multifunctional rotary valve, a 6-port switching valve, and a three-way valve. The multi functional rotary valve comprises a stator having a plurality of ports for connecting fluid lines thereto, and a rotor coaxially rotatable with respect to the stator. The rotor comprises channels. The ports on the stator are arranged in a plurality of groups. The stator has at least one group of ports. Each group has at least three ports, among which one of the ports is used as common port. The common port may be used as an outlet or inlet for fluid flowing through a component connected to the two non-common ports in the same group, for example. The stator also comprises a central port. The central port may be used to connect with a pump to deliver solvents to other ports on the stator or may be used as a common outlet for other ports on the stator. The channels on the rotor are arranged such that by rotating the rotor the common port in each group of the ports of the stator can be connected with one of the other two ports in the group or to the port located in the center of the stator and that when the central port is connected to one non-common port, the other non-common port in the same group is always connected to the common port in that group.
Stator 11 and rotor 12 are received in a housing 14. Stator 11 is rigidly fixed to upper annular sidewalls 27 of housing 14 by a plurality of fasteners 15. Fasteners 15 may for example be screws which are received in corresponding threaded holes 28 in upper annular sidewalls 27. In other embodiments, stator 11 may be fixed to housing 14 in any other manner known in the art.
Rotor 12 is held within housing 14 by stator 11. Rotor extension 12E extends through an opening 29 at the bottom of housing 14. Rotor 12 is coaxially rotatable relative to stator 11 about axis 16 of stator 11. Some embodiments may include a washer 17 to facilitate a sealed connection and relative rotation between rotor upper face 12F and stator lower face 4. Some embodiments may also include a washer 13 to facilitate a sealed connection and relative rotation between rotor 12 and housing 14.
Channels on rotor 2 are formed with two types on interface 260 of rotor 2. The first channel radiates from the axial center of rotor 2 or center of interface 260. In the illustrate embodiment, the first channel is linear groove 251. As shown in
Two second channels are arranged with the first channel in between and in a concentric manner around the center of interface 260 of rotor 2 along a path corresponding to imaginary circle 261 when rotor 2 and stator 1 are fitted together (see
In
Accordingly, by selectively rotating rotor 2 coaxially with respect to stator 1, each port can have one of four statuses: 1) in fluid communication with a first one of their adjacent ports in the same group; 2) in fluid communication with a second one of their adjacent ports in the same group; 3) in fluid communication with central port 100, or 4) blocked from all the other ports.
Herewith a typical sample preparation procedure for two-stage online SPE is used as an example to explain the working principle of the present apparatus.
The first step is eluting SPE column C1 with solvent S1 to wet the sorbent bed. The working steps are 1) rotating valve 200 to connect syringe pump SP with solvent S1 and drawing solvent into the syringe pump SP (
The second step is loading sample SA to SPE column C1 in reversed direction. The working steps are 1) rotating valve 200 to connect syringe pump SP with sample SA via adapter P1 and drawing sample fluid into the syringe pump (
The third step is washing away interfering components from sample matrices from SPE column C1 using solvent S2. The working steps are 1) rotating valve 200 to connect syringe pump with solvent S2 and drawing solvent to the syringe pump; 2) rotating valve 200 to connect syringe pump SP with SPE column 01 via adapter P2 and pushing solvent S2 to SPE column C1 in a normal direction (
The fourth step is washing down the targeted components from SPE column C1 using solvent S3 and collecting the fraction F. The working steps are 1) rotating valve 200 to connect syringe pump SP with solvent S3 and drawing solvent into the syringe pump; 2) rotating valve 200 to connect syringe pump SP with SPE column C1 via adapter P2, switching valve 700 to connect adapter P3 with the effluent of SPE column C1 via adapter CS and pushing solvent S3 to SPE column C1 in a normal direction (
The fifth step is transferring the collected fraction to SPE column C2 for further cleanup and enrichment. The working steps are: 1) rotating valve 200 and valve 700 to connect syringe pump SP with collected fraction F via adapter P3 and drawing fraction fluid into syringe pump (
The sixth step is washing the SPE column C2 to remove more interfering components using solvent S2. The working steps are: 1) rotating valve 200 to connect syringe pump SP with solvent S2 and drawing solvent S2 into syringe pump SP; 2) rotating valve 200 again to connect syringe pump SP with SPE column C2 and pushing the solvent to SPE column C2 in a normal direction (
The seventh step is transferring targeted components from SPE column C2 to LC column C3 for LC analysis. The working step is rotating valve 500 to connect SPE column C2 with LC pump LP and LC column C3 (
While a number of exemplary aspects and embodiments of the invention have been discussed above, those of skill in the art will recognize certain modifications, permutations, additions and sub-combinations thereof. The scope of the claims should not be limited by the preferred embodiments set forth in the examples, but should be given the broadest interpretation consistent with the description as a whole.