High-pressure fluid processing apparatus for liquid chromatography

Abstract

The present invention provides a high-pressure fluid apparatus. The high-pressure fluid apparatus comprises a housing, a tube, and a stationary phase medium. The housing includes a first component having a first contact surface and a second component having a second contact surface. The first and second components are fixable to each other by their opposing contact surfaces, forming a contact portion between them. The housing includes a channel extending along the contact portion through the first component to the contact portion. The tube extends along the channel. The tube defines a lumen having a substantially circular cross-section with a lumen diameter of 1000 μm or less. The stationary phase medium is positioned in the lumen. The stationary phase medium is suitable for chromatographic separation of compounds present in a liquid sample.

Description

Technical Field

[0001] This application claims the benefit and priority of U.S. Provisional Application No. 63 / 463,413, filed May 2, 2023, entitled "High Pressure Fluid Handling Device for Liquid Chromatography," which is hereby incorporated by reference in its entirety.

[0002] The subject matter disclosed herein generally relates to liquid chromatography systems, and more specifically, to fluid handling devices for transporting fluids in high pressure and ultra-high pressure liquid chromatography applications. The fluid handling device may be particularly adapted for microscale and / or nanoscale chromatography applications to provide a liquid chromatography column having an inner diameter of 1000 μm or less.

Background Art

[0003] High-performance liquid chromatography (HPLC) is a known technique for separating the components of a sample. The liquid used to carry the sample through the chromatography system is called the "mobile phase." The components of the sample may be separated in a "column," which is typically a tube filled with particulate material having varying degrees of affinity for the various components. The packing material is called the "stationary phase" and is usually a silica or polymer-based medium that may be coated with chemical functional groups to promote the desired chemical affinity with the components of the sample.

[0004] Conventional HPLC instruments typically use columns made from hard stainless steel conduits. Other inert materials, such as various polymers, are also used in the construction of chromatography columns. Typical HPLC columns have an inner diameter in the range of approximately 1–10 mm and a stationary phase packing length in the range of approximately 5–25 cm. Recently, ultra-high performance liquid chromatography (UHPLC) has been found to have important applications in analyzing smaller sample volumes, with shorter test durations, and with higher sensitivity, but at considerably higher pressures (>10,000 psi) than conventional HPLC. UHPLC systems are often operated as microscale, capillary-scale, or nanoscale HPLC, with mobile phase flow rates ranging from approximately 100 nL / min to 100 μL / min and column inner diameters between 25–1000 μm. The advantages resulting from this reduction in size, though not all, include reduced solvent consumption (both environmental and economic benefits), improved sensitivity, reduced stationary phase usage, improved compatibility with mass spectrometry, and analysis of small sample volumes. Because sufficiently high-quality stainless steel conduits are typically unavailable in these dimensions, alternative materials such as fused silica may be used in the fabrication of the columns. However, such materials are brittle, flexible, and require special handling care.

[0005] One proposed solution is a patterned substrate, or "chip"-based design, in which channels of desired dimensions are created within a substrate similar to an electrical equivalent such as a computer chip. In chip-based designs, the channels are typically formed on the substrate by chemical etching, lithography, or CNC machining and are usually provided in square, rectangular, D-shaped, trapezoidal, or other polygonal cross-sectional shapes. These non-circular cross-sectional shapes perform worse than those expected in circular cross-sectional shapes, particularly at the "corners" or cracks of the non-circular cross-section, due to flow field asymmetry and the non-uniform density of the packed stationary phase, resulting in the spread of the sample plug and dispersion of the relevant components. Attempts to create circular cross-sectional channels have so far required aligning and joining two different substrate layers, minimizing or eliminating displacement and ensuring gap- or crack-free layers. This approach has not consistently proven feasible.

[0006] Conventional patterned channels have the drawback of abrupt changes in direction within the substrate, such as 90° bends, which tends to hinder consistent stationary phase column packing and cause band broadening in chromatographic applications. These abrupt changes in direction often occur at the contact points between substrate layers where the mobile phase and sample are received from the tip inlet and / or delivered to the tip outlet. [Overview of the project] [Means for solving the problem]

[0007] According to one embodiment, a high-pressure fluid apparatus is provided. The high-pressure fluid apparatus includes a housing, a tube, and a stationary phase medium. The housing includes a first component having a first contact surface and a second component having a second contact surface. The first component and the second component are fixed to each other by the first and second contact surfaces, which are in a relationship to each other so as to form a contact portion between them. The housing includes a channel extending along the contact portion through the first component to the contact portion. The tube extends along the channel. The tube defines a lumen having a substantially circular cross-section with a lumen diameter of 1000 μm or less. The stationary phase medium is positioned in the lumen. The stationary phase medium is suitable for chromatographic separation of compounds present in a liquid sample.

[0008] In another embodiment, a high-pressure fluid apparatus is provided. The inlet is configured to receive a liquid sample. The housing includes a first component having a first contact surface and a channel extending through the first contact surface. The housing includes a second component having a second contact surface. The first and second components are fixed to each other by the first and second contact surfaces, which are in a relationship of opposing relations, so as to form a contact portion between them. A tube is positioned within the channel of the first component. The tube defines a lumen having a substantially circular cross-section. A stationary phase medium is disposed within the lumen of the tube. The stationary phase medium is suitable for chromatographic separation of compounds present in the liquid sample. An outlet port is fluidically coupled to the inlet port via the tube.

[0009] In another embodiment, a method for forming a high-pressure fluid apparatus is provided. A first component having a first contact surface is provided. A channel is etched onto the first contact surface of the first component. A tube is positioned within the channel. The tube includes a lumen having a substantially circular cross-section with a diameter of 1000 μm or less. A second component having a second contact surface is fixed to the first component. The tube is filled with a stationary phase medium. The stationary phase medium is suitable for separating compounds present in a liquid sample by chromatography. [Brief explanation of the drawing]

[0010] [Figure 1] These are isometric views of high-pressure fluid processing devices according to several embodiments. [Figure 2] This is a schematic diagram of a liquid chromatography system including a high-pressure fluid processing apparatus, according to several embodiments. [Figure 3] These are isometric views of the first and second components of a high-pressure fluid processing apparatus according to several embodiments. [Figure 4] These are isometric bottom views of a first and second component of a high-pressure fluid processing apparatus according to several embodiments. [Figure 5] This is an isometric cross-sectional view of a high-pressure fluid processing apparatus according to several embodiments. [Figure 6] This is an enlarged isometric cross-sectional view of a rounded bend in a pipe in a high-pressure fluid processing apparatus according to several embodiments. [Figure 7] This is a cross-sectional view of pipes and channels extending through a high-pressure fluid processing apparatus according to several embodiments. [Figure 8] This is an isometric bottom view of a first component of a high-pressure fluid processing device according to several embodiments. [Figure 9] This is a cross-sectional side view of a high-pressure fluid apparatus having a pipe terminating at an outlet on the outer surface of the high-pressure fluid apparatus, according to several embodiments. [Figure 10] This is a cross-sectional side view of a multilayer tube of a high-pressure fluid processing device according to several embodiments. [Figure 11A] This is a cross-sectional view of a high-pressure fluid processing apparatus having a membrane positioned above an outlet to form a cavity, according to several embodiments. [Figure 11B] This is a cross-sectional view of a high-pressure fluid processing apparatus having a stationary phase held in a cavity, according to several embodiments. [Figure 11C] This is a cross-sectional view of a high-pressure fluid processing apparatus in which a membrane has been removed to expose a cavity, according to several embodiments. [Figure 11D]This is a cross-sectional view of a high-pressure fluid processing apparatus having porous frit installed in a cavity to hold a stationary phase inside a pipe, according to several embodiments. [Figure 12A] These are chromatograms of fluid processing devices filled with a stationary phase at 1,500 psi, according to several embodiments. [Figure 12B] These are chromatograms of fluid processing devices filled with a stationary phase at 16,500 psi, according to several embodiments. [Figure 13] This is a flowchart of a method for forming a high-pressure fluid processing apparatus according to several embodiments. [Figure 14] This is a flowchart of a method for filling the tubes of a high-pressure fluid processing device, according to several embodiments. [Modes for carrying out the invention]

[0011] This disclosure describes a fluid apparatus for use in liquid chromatography. The high-pressure fluid apparatus comprises a housing having a small-diameter fluid passage extending through the housing. The small-diameter fluid passage is configured to withstand high pressure (i.e., includes thick side walls and / or is trapped by the housing to support the high-pressure load). The small-diameter fluid passage is configured for use in microscale, capillary-scale, and / or nanoscale liquid chromatography, and its cross-sectional diameter is 1000 μm or less. The small-diameter fluid passage extends along the length of the high-pressure fluid apparatus, and the length of the small-diameter fluid passage is at least 100 times its diameter. Because the fluid passage has a high aspect ratio (length to diameter) and / or its diameter is microscale / nanoscale, it is not practical to drill holes in the housing.

[0012] To provide a small-diameter fluid passage extending along the length of a high-pressure fluid apparatus configured to withstand high-pressure loads, the housing includes a first component having a first contact surface and a second component having a second contact surface. The first and second components are fixed together at their opposing contact surfaces so as to form a contact portion between them. The housing includes a channel extending through the first component along the contact portion. A tube defining a small-diameter fluid passage having a substantially circular cross-section is positioned in the channel. The fluid passage is filled with a stationary phase medium suitable for chromatographic separation of liquid samples.

[0013] For example, if a channel extends through both a first and a second component (e.g., the first component contains a semicircular channel and the second component contains a semicircular channel), then near-perfect alignment between the first and second components would be required. If the alignment is off by even a micrometer, sharp edges will form along the fluid passage, impairing the liquid chromatography process.

[0014] FIG. 1 is an isometric view of a high-pressure fluid processing apparatus 10 according to some embodiments. The high-pressure fluid processing apparatus 10 includes a housing 12 having a first component 14 and a second component 16 that form a contact portion 38 therebetween. The high-pressure fluid processing apparatus 10 includes an inlet block 18 fixed to the first component 14 and / or the second component 16 via one or more fasteners 24. The inlet block 18 includes an inlet tube receptacle 19 configured to receive a liquid sample and / or configured to be fixed to an inlet tube (or conduit) for transporting the liquid sample. The high-pressure fluid processing apparatus 10 includes an outlet block 22 fixed to the first component 14 and / or the second component 16 via one or more fasteners 24. The outlet block 22 includes an outlet tube receptacle 23 configured to discharge a liquid sample and / or configured to be fixed to an outlet tube (or conduit) for transporting the liquid sample. In some embodiments, the inlet tube receptacle 19 and the outlet tube receptacle 23 are configured to engage and receive pipe fittings that can effectively establish a fluid connection. The inlet tube receptacle and the outlet tube receptacle may be provided with threads for threadably engaging with the pipe fittings.

[0015] The high-pressure fluid processing apparatus 10 may be used for various fluid transport applications. In some embodiments, the high-pressure fluid processing apparatus 10 may be particularly well-suited for liquid chromatography applications such as high-performance liquid chromatography (“HPLC”) and / or ultra-high performance liquid chromatography (“UHPLC”). For example, the high-pressure fluid processing apparatus 10 is configured to receive a liquid sample through the inlet block 18. The liquid sample passes through the high-pressure fluid processing apparatus 10 and exits the high-pressure fluid processing apparatus 10 through the outlet block 22. The high-pressure fluid processing apparatus 10 may be referred to as a “chip” (for chip-based chromatography applications) or, more generally, a “manifold”.

[0016] FIG. 2 is a schematic diagram of a high-pressure liquid chromatography system 102 including a high-pressure fluid treatment device 10 according to some embodiments. The high-pressure liquid chromatography system 102 includes a solvent reservoir 104 (and / or an eluent reservoir), a degasser 106 for degassing the liquid solvent / eluent, a pump such as a high-pressure pump 108 that pumps the degassed liquid solvent / eluent to an injection valve 110, and a sample from a sample container or an automatic sample injector 112 is injected into the solvent stream / eluent stream and delivered to a column 114 for chromatographic separation of the sample. Next, the plug of the sample separated by chromatography may be analyzed by a detection device 116 such as a mass spectrometer or an ultraviolet / visible spectrum detector. Data analysis is performed by a data acquisition device 118 such as a computer, and the waste liquid may be collected in a waste reservoir 120.

[0017] In some embodiments, the high-pressure liquid chromatography system 102 optionally includes additional or different components, such as a flow cell for optical analysis, an in-line degasser, a sample loop, etc., as are well known in the art. The high-pressure fluid treatment device S10, described with reference to the figures, includes, according to some embodiments, a portion of the chromatography system 102 between the injection valve 110 and the analyzer 116. The high-pressure fluid treatment device 10 may include, but is not limited to, a flow cell for optical detection, a heating or cooling element for controlling temperature, and / or an emitter tip for use in mass spectrometry, does not require an external tube, includes additional elements, consists of additional elements, or may be directly integrated with additional elements.

[0018] Figure 3 is an isometric view of a first component 14 and a second component 16 of a high-pressure fluid processing apparatus 10 according to several embodiments. The first component 14, according to some embodiments, includes an outer surface 15, an inlet opening 30, an outlet opening 32, one or more mounting containers 26, and one or more alignment features 28. The high-pressure fluid processing apparatus 10 is configured to facilitate fluid sealing between an inlet pipe (not shown) and the inlet opening 30, and between an outlet pipe (not shown) and the outlet opening 32. In some embodiments, an inlet block 18 and an outlet block 22 are fixed to the first component 14 to prevent fluid from escaping between the respective inlet block 18 and outlet block 22 and the first component 14. Thus, the inlet and outlet pipes, fixed to each inlet block 18 and outlet block 22 with couplings that prevent fluid leakage from the containers 19, 23, transport the fluid (i.e., mobile phase) to the inlet opening 30 and out of the outlet opening 32.

[0019] Figure 4 is an isometric view of a first component 14 and a second component 16 of a high-pressure fluid processing apparatus 10 according to several embodiments. The second component 16, according to some embodiments, includes an outer surface 17, one or more mounting containers 26, and one or more alignment features 28. One or more mounting containers 26 positioned on the first component 14 align with one or more mounting containers 26 positioned on the second component 16 (as shown in Figure 3). Similarly, one or more alignment features 28 positioned on the first component 14 (as shown in Figure 3) align with one or more alignment features 28 positioned on the second component 16. In some embodiments, one or more mounting containers 26 and / or one or more alignment features 28 extend through the first component 14 and the second component 16 of the housing 12 to align and fix the first component 14 and the second component 16 together. For example, a fastener 24 extends through one or more mounting receivers 26 to connect the first component 14 and the second component 16 together, and a stud / pin (not shown) extends through one or more alignment features 28 to align the first component 14 with respect to the second component 16.

[0020] In some embodiments, the first component 14 and the second component 16 can be aligned and fastened together without the mounting receiver 26 and / or alignment feature 28. For example, the force required to maintain fluid sealing is provided by external means (not shown), such as a clamping mechanism. In this embodiment or other embodiments, the high-pressure fluid processing apparatus 10 may be configured to facilitate fluid sealing directly between adjacent components of the chromatography system 102, for example, between the high-pressure fluid processing apparatus 10 and the injection valve 110, and / or between the high-pressure fluid processing apparatus 10 and the detection device 116, with or without the use of one or more inlet and outlet pipes.

[0021] Figure 4, an isometric bottom view, shows the outer surface 17 of the second component 16. Figures 3 and 4 show a housing 12 including the first component 14 and the second component 16, the housing 12 may comprise multiple layers of the same or different materials to form the overall “tip” or “manifold” of the high-pressure fluid processing apparatus 10. For the purposes of this application, the housing 12 may be configured to define a contact portion 38 between two contact surfaces. In the illustrated embodiment, each of the first component 14 and the second component 16 includes a contact surface, the first component 14 includes a first contact surface 34, and the second component 16 includes a second contact surface 36. The first contact surface 34 and the second contact surface 36 engage (or abut) with each other at the contact portion 38.

[0022] In some embodiments, the first component 14 and the second component 16 can be fixed to each other at a first contact surface 34 and a second contact surface 36 that are in a mutually opposing relationship, forming a contact portion 38. For example, the first component 14 and the second component 16 can be fixed to each other at a first contact surface 34 that engages with the second contact surface 36 in order to hold a pipe 40 between them and maintain the overall integrity of the fluid processing apparatus 10. The engagement between the first contact surface 34 and the second contact surface 36 is configured to avoid misalignment between the first component 14 and the second component 16. In some embodiments, an alignment feature 28 runs throughout the housing 12 and minimizes / eliminates a gap or sharp feature between the first contact surface 34 and the second contact surface 36. In some embodiments, the first contact surface 34 and the second contact surface 36 are fixed together by brazing, laser welding, adhesive, mechanical fastening, etc.

[0023] Figure 5 is an isometric cross-sectional view of a high-pressure fluid processing apparatus 10 according to several embodiments. According to some embodiments, the high-pressure fluid processing apparatus 10 includes a channel 20 extending through a first component 14 between an inlet opening 30 and an outlet opening 32. The channel 20 is located on the outer surface 15 of the first component 14 at the inlet port 30 and the outlet port 32. In some embodiments, the channel 20 extends from the outer surface 15 through the first component 14 to a contact surface 34. The channel 20 is located along the contact surface 34, and in some embodiments, the channel 20 is formed by milling, cutting, laser etching, chemical etching, lithography, CNC machining, etc. of the contact surface 34. In other embodiments, the first housing 14 is formed by a mold or extruder, with the channel 20 formed in the mold / extruder. In the illustrated embodiment, the channel 20 is formed only in the first component 14 (but not through the second component 16), although it is assumed that the channel 20 may be formed in one or more layers of the high-pressure fluid processing apparatus 10. The channel 20 may form one or more through holes passing through at least one of the first component 14 and the second component 16, and / or recesses or grooves on one or more surfaces of the first component 14 and the second component 16.

[0024] Figure 6 is an enlarged isometric cross-sectional view of a high-pressure fluid processing apparatus 10 according to several embodiments. The high-pressure fluid processing apparatus 10 includes a tube 40 positioned within a channel 20 and a rounded bend 21 of the channel 20. The tube 40 is positioned within the channel 20 of the high-pressure fluid processing apparatus 10 and extends through and / or along one or more of the first components 14 and the second components 16. In some embodiments, the tube 40 extends from an inlet opening 30 to an outlet opening 32. In doing so, the tube 40 extends along the channel 20, through the first components 14, and along the contact portion 38.

[0025] The rounded bend 21 of the channel 20 is configured to transition the position of the channel 20 from the outer surface 15 to the contact surface 34 without sharp edges or abrupt changes in direction that would disrupt the fluid flow characteristics. In some embodiments, the rounded bend 21 may be between 5° and 180°, and the illustrated embodiment has a 90° rounded bend 21. In some embodiments, the rounded bend 21 may be between 60° and 120°.

[0026] In some embodiments, the tube 40 is manufactured from a material and formed to be able to establish and maintain the lumen diameter described below while following one or more rounded bends 21. In some embodiments, the tube 40 is manufactured from biocompatible materials such as stainless steel, titanium, polyetheretherketone, and combinations thereof, for example, a stainless steel conduit internally coated with polyetheretherketone. Other metals, metal alloys, polymer materials, and combination materials are considered useful for manufacturing the tube 40, and furthermore, coated versions of the tube 40 are also envisioned, in which one or more surfaces of the tube 40 are coated, thereby imparting beneficial properties such as chemical compatibility and bioinertness.

[0027] Figure 7 is a cross-sectional view of a tube 40 and a channel 20 extending through a high-pressure fluid processing apparatus 10 according to several embodiments. The tube 40 defines a lumen 42. In some embodiments, the lumen 42 has a substantially circular cross-sectional shape. In some embodiments, the lumen 42 has a circular cross-section, and the diameter dimensions along the orthogonal radial axes in the lumen 42 are within 5% of each other, preferably within 3% of each other, and more preferably within 2% of each other. Thus, the precision of the configuration of the channel 20 is not important, as long as the tube 40 is positioned in the channel 20 to provide the desired fluid connection. As mentioned above, the arrangement of the fluid passage to have a circular cross-section is most desirable to facilitate the expected flow pattern and its influence on the eluent containing the sample. Therefore, the tube is configured to maintain a substantially circular lumen cross-section when positioned in the channel 20.

[0028] In some embodiments, the lumen 42 defines microscale, capillary scale, and / or nanoscale diameters configured for use in HPLC and / or UHPLC. According to some embodiments, the lumen 42 has a lumen diameter of 1000 μm or less. In some embodiments, the lumen 42 has a lumen diameter between 25 and 1000 μm, in some embodiments, the lumen 42 has a lumen diameter between 25 and 500 μm, in some embodiments, the lumen 42 has a lumen diameter between 25 and 250 μm, and / or in some embodiments, the lumen 42 has a lumen diameter between 25 and 150 μm.

[0029] Figure 8 is an isometric view of a first component 14 of a high-pressure fluid processing apparatus 10 according to several embodiments. The first component 14 includes a contact surface 34 with a channel 20 formed therein. A tube 40 is positioned within the channel 20. In some embodiments, the tube 40 is brazed to the channel 20. For example, the tube 40 is brazed into the channel 20 such that the gap between the tube 40 and the channel 20 is filled with brazing metal. In other embodiments, the gap between the tube 40 and the channel 20 is filled with adhesive by fixing the tube 40 to the channel 20 with adhesive.

[0030] In some embodiments, the channel 20 is entirely formed in the first component 14. That is, the contact surface 36 of the second component 16 does not include any recesses or channels that form part of the channel 20. For example, as shown in Figure 7, the contact surface 36 of the second component 16 is flat. In some embodiments, the tube 40 is entirely disposed within the channel 20 of the first component 14. That is, no portion of the tube 40 extends into any recess, channel, or other concave feature of the second component 16.

[0031] In some embodiments, positioning the channel 20 only in the first component 14 (and not in the second component 16) is beneficial because it does not require perfect alignment between the first component 14 and the second component 16. For example, if the channel 20 extends through both the first component 14 and the second component 16 (e.g., the first component 14 contains a semicircular channel and the second component 16 contains a semicircular channel), then near-perfect alignment is required between the first component 14 and the second component 16. If the alignment is off by even a micrometer, a sharp edge will be formed along the fluid passage, which will interfere with the liquid chromatography process. Therefore, positioning the tube 40 only in the first component 14 provides a substantially circular flow path without requiring near-perfect alignment along the entire length of the high-pressure fluid processing apparatus 10.

[0032] In some embodiments, positioning the tube 40 between the first component 14 and the second component 16 is beneficial because the first component 14 and the second component 16 structurally support the side walls of the tube 40. The flow path of the high-pressure fluid processing apparatus 10 may need to withstand high pressure (e.g., pressures between 1,500 psi and 16,500 psi and / or pressures in the range of 20,000 psi). The first component 14 and the second component 16 abut the tube 40 and provide the structural support necessary to withstand the high pressure (e.g., in the range of 20,000 psi) of the HPLC and UHPLC systems.

[0033] Figure 9 is a cross-sectional side view of a high-pressure fluid processing apparatus 10 having a pipe 40 terminating at an outlet 70 on the outer surface 25 of the high-pressure fluid processing apparatus 10, according to several embodiments. The high-pressure fluid processing apparatus 10 includes an outlet 70 located on the outer surface 25 of the housing 12. Furthermore, it will be readily apparent to those skilled in the art that the channel 20 may begin and / or terminate on any surface of the housing 12, and that in some embodiments, the beginning and ending surfaces may be the same surface.

[0034] In some embodiments, the tube 40 has a diameter d, and the channel 20 includes a height (or depth) h. The height h of the channel 20 is greater than or equal to the diameter d of the tube 40. This configuration is advantageous because the tube 40 is received within the channel 20, which is located only within the first component 14. That is, the channel 20 does not extend beyond the contact surface 36 of the second component 16, and / or no portion of the tube 40 is received within the features of the second component 16.

[0035] Figure 10 is a cross-sectional side view of a multilayer tube 80 of a high-pressure fluid processing apparatus 11 according to several embodiments. According to several embodiments, the high-pressure fluid processing apparatus 11 includes any and / or all of the features of the high-pressure fluid processing apparatus 10 described above. According to several embodiments, the multilayer tube 80 includes an inner wall 72 and an outer wall 74. In some embodiments, the multilayer tube 80 is manufactured from biocompatible materials such as stainless steel, titanium, polyetheretherketone, and combinations thereof, for example, a stainless steel conduit internally coated with polyetheretherketone. Other metals, metal alloys, polymer materials, and combination materials are considered useful for manufacturing the multilayer tube 80, and furthermore, coated versions of the tube 80 are also envisioned, in which one or more surfaces of the multilayer tube 80 are coated, thereby imparting beneficial properties such as chemical compatibility and bioinertness. Furthermore, those skilled in the art will readily understand that the multilayer tube 80 may include any number of layers and / or materials. In some embodiments, an outlet frit 76 is located within the multilayer tube 40 to prevent the stationary phase material from leaving the high-pressure processing apparatus 10. In some embodiments, the outer wall 74 is formed of brazed metal that fills the gap between the pipe 40 and the channel 20. According to some embodiments, the multilayer pipe 80 is configured to be positioned on a rounded bend 81.

[0036] Figures 11A to 11D are cross-sectional views illustrating exemplary embodiments of filling (or "filling") a lumen 92 with a stationary phase 60, according to several embodiments. For example, Figure 11A is a cross-sectional view showing a filter 50 positioned above the outlet opening 32 and fixed to the housing 12 for the purpose of filling the lumen 92. The filter 50 is impermeable to the stationary phase (not shown in Figure 11A), i.e., the filter 50 contains pores smaller than the particle size of the stationary phase. The filter 50 is configured to allow fluid (i.e., liquid-filled slurry) to pass through the filter 50 once the stationary phase 60 is filled and held in the tube 90. The high-pressure fluid processing apparatus 10 includes a cavity 52 located adjacent to the outlet 32 ​​and between the ends 96 of the tube 90. In some embodiments, the tube 90 does not extend to the outer surface 15 of the first housing 14; instead, the cavity 52 is formed in the space between the tube 40 and the outer surface 15 (within the channel 20).

[0037] Figure 11B shows the lumen 92 of a tube 90 filled with a stationary phase 60 in some embodiments. The stationary phase 60 is pressure-driven into the lumen 92 of the tube 90 through an inlet 30 (not shown in Figure 11B) and fills the lumen 42. The stationary phase 60 is biased in the flow direction from the inlet 30 (not shown in Figure 11B) to the outlet 32, i.e., as indicated by the arrow 94. In some embodiments, a filter 50 and / or block 54 located on the outer surface 15 of the first housing 14 allows for high filling pressure by preventing the escape of the stationary phase through the outlet 32, even under high pressure. The cavity 52 is filled with the stationary phase 60.

[0038] In some embodiments, the stationary phase 60 is packed into the lumen 92 at a pressure between 1,500 psi and 20,000 psi. As recognized by those skilled in the art, improved performance in HPLC and / or UHPLC can result from higher packing pressures that require a correspondingly higher pressure capacity. Conventional stationary phase packing can be performed between 1,500 psi and 2,000 psi to pack the stationary phase into the lumen 92. However, stationary phase packing performed between 10,000 psi and 20,000 psi may yield better separation results. In some embodiments, when the stationary phase 60 is packed at a high pressure (e.g., between 5,000 psi and 20,000 psi), a semipermeable filter 50 holds the stationary phase 60 in the lumen 92, and positioning the filter 50 at the outlet allows for higher packing pressures of the stationary phase 60.

[0039] For example, Figures 12A and 12B show chromatograms obtained under similar conditions through a fluid processing apparatus 10 having a tube length of 150 mm and a lumen diameter of 200 μm. The chromatograms show the separation of thiourea, acetophenone, propiophenone, and butyrophenone in 45% ACN:55% H2O at a rate of 2 μl / min using a 2.7 μm surface porous 160 Å C18 stationary phase. Figure 12A shows the results for a stationary phase packed at 1,500 psi to 2,000 psi, while Figure 12B shows the results for a stationary phase packed at 16,500 psi. The significantly higher theoretical plate count in Figure 12A compared to Figure 12B indicates improved chromatographic performance achievable by higher stationary phase packing pressures, which are facilitated by the packing techniques described herein.

[0040] Figure 11C shows the removal of the filter 50 from the outer surface 15 in some embodiments. Removing the filter 50 (and block 54) exposes the cavity 52, allowing for visual inspection of the outlet 32 ​​and confirmation of whether the stationary phase 60 is present at the outlet 32. In some embodiments, the stationary phase 60 is removed from the cavity 52 after the filter 50 has been removed.

[0041] Figure 11D shows a porous frit 62 (i.e., a porous plug) placed in a cavity 52 to hold the stationary phase 60 in a tube 90, according to several embodiments. The porous frit 62 is impermeable to the stationary phase but permeable to a liquid sample and / or liquid solvent (or eluent). In some embodiments, the porous frit 62 is made from stainless steel, titanium, stainless steel-nickel alloy and / or polyetheretherketone. It will be readily apparent to those skilled in the art that the arrangements shown in Figures 11A to 11D can be used in conjunction with an inlet, e.g., inlet 30, or applied separately from the inlet.

[0042] Figure 13 is a flowchart of a method 1300 for forming a high-pressure fluid processing apparatus according to several embodiments. In step 1310, the method 1300 includes providing a first component. The first component includes any and / or all of the features of the first component 14 described above in any of Figures 1 to 11D.

[0043] Step 1320 of Method 1300 includes forming a channel within the first contact surface of the first component. The channel includes any and / or all of the features of the channel 20 described above. In some embodiments, etching the channel 20 includes any of the various known techniques, including laser etching, chemical etching, lithography, CNC machining, etc.

[0044] Step 1330 of Method 1300 includes positioning the tube within the channel. The tube includes any and / or all of the features of the tube 40 described above. In some embodiments, the tube 40 is fully positioned within the channel 20. That is, the depth of the channel 20 is greater than or equal to the outer diameter of the tube 40 so that the tube 40 is fully positioned within the first component 14. In some embodiments, the second contact surface 36 is substantially planar, and the tube 40 does not intersect with the substantially planar second contact surface 36. In some embodiments, step 1330 includes brazing the tube 40 within the channel 20 such that the gap between the tube 40 and the channel 20 is filled with brazing metal. In some embodiments, step 1330 includes filling the gap between the tube 40 and the channel 20 with epoxy resin and / or other adhesive or brazing material.

[0045] Step 1340 of the method 1300 includes fixing a second component having a second contact surface to a first component. The second component includes any and / or all of the features of the second component 16 described above. In some embodiments, the first component 14 is fixed to the second component 16 via fasteners 24 and / or alignment features 28. In some embodiments, the first component 14 is fixed to the second component 16 by brazing, welding, fastening, etc.

[0046] Step 1350 of Method 1300 includes filling the tube with a stationary phase medium. The stationary phase medium includes any and / or all of the features of the stationary phase 60 described above. In some embodiments, the stationary phase 60 is filled into the lumen 42 at a filling pressure of at least 1,500 psi. In some embodiments, the stationary phase 60 is filled into the lumen 42 at a filling pressure of at least 2,000 psi. In some embodiments, the stationary phase 60 is filled into the lumen 42 at a filling pressure of at least 5,000 psi. In some embodiments, the stationary phase 60 is filled into the lumen 42 at a filling pressure of at least 10,000 psi. In some embodiments, the stationary phase 60 is filled into the lumen 42 at a filling pressure between 10,000 psi and 20,000 psi. For the purposes of this specification, the term “filling pressure” means the fluid pressure that drives the stationary phase 60 into the lumen 42. In some embodiments, the fluid pressure is the pressure of the liquid filling slurry. In some embodiments, the liquid-filled slurry may be pressurized via a pressurized gas volume. As will be recognized by those skilled in the art, performance improvements in HPLC and / or UHPLC may result from higher filling pressures that require a correspondingly higher pressure capacity.

[0047] Figure 14 is a flowchart of Method 1400 for filling the tubes of a high-pressure fluid processing apparatus according to several embodiments. In some embodiments, Method 1300 includes any and / or all of the steps, features, or processes shown and described with reference to Figures 11A to 11D. Step 1410 includes Method 1400 providing a high-pressure fluid processing apparatus. The high-pressure fluid processing apparatus includes any and / or all of the features of the high-pressure fluid processing apparatus 10 described above in Figures 1 to 11D.

[0048] Step 1420 of Method 1400 includes positioning filters at the inlet and / or outlet. The inlet / outlet includes any and / or all of the features of the inlet 30 and outlet 32 ​​described above. The filter (e.g., filter 50) can be positioned on the outer surface 15 to provide an impermeable film to the stationary phase medium. Step 1430 of Method 1400 includes filling the tube with the stationary phase medium. In some embodiments, the stationary phase 60 is filled into the lumen 42 of the tube 40 at a pressure between 1,500 psi and 20,000 psi. As will be recognized by those skilled in the art, improved performance of HPLC and / or UHPLC may result from higher filling pressures that require a correspondingly higher pressure capacity.

[0049] In step 1440, method 1400 includes removing the filter from the inlet / outlet. The filter 50 can be removed from the outer surface 15, thereby exposing the inlet 30 and / or outlet 32 ​​so that the presence of the stationary phase 60 can be visually confirmed. In step 1450, the stationary phase medium is removed from the cavity. In some embodiments, the stationary phase 60 is removed from the cavity 52 after the removal of the filter 50. In step 1460, a porous frit (or filter) is inserted into the cavity. In some embodiments, a porous frit 62 is placed in the cavity 52 to hold the stationary phase 60 inside the tube 40. The porous frit 62 is impermeable to the stationary phase 60 but permeable to the liquid sample and / or liquid solvent (or eluent).

[0050] The present invention is described with reference to exemplary embodiments, but it will be understood by those skilled in the art that various modifications can be made and equivalents can be substituted for elements without departing from the scope of the invention. Furthermore, many modifications can be made to adapt the teachings of the invention to specific situations or materials without departing from the essential scope of the invention. Thus, the invention is not limited to the specific embodiments disclosed, but is intended to include all embodiments that fall within the scope of the appended claims.

Claims

1. A housing having a first component having a first contact surface and a second component having a second contact surface, wherein the first component and the second component are fixable to each other by the first and second contact surfaces which are in a relationship to each other so as to form a contact portion between them, and the housing has a channel extending along the contact portion through the first component to the contact portion, A tube extending along the channel, defining a lumen having a substantially circular cross-section with a lumen diameter of 1000 μm or less, A high-pressure fluid apparatus comprising a stationary phase medium in the lumen, which is suitable for separating compounds present in a liquid sample by chromatography.

2. The high-pressure fluid apparatus according to claim 1, wherein the first component and the second component are fixed to each other by the first and second contact surfaces which are sealed and engaged.

3. The high-pressure fluid processing apparatus according to claim 1, wherein the channel has an inlet opening and an outlet opening formed in the first component.

4. The high-pressure fluid processing apparatus according to claim 3, wherein the pipe extends from the inlet opening to the outlet opening.

5. The high-pressure fluid apparatus according to claim 1, wherein the tube is sealed and engaged with the wall of the first component defining the channel to prevent liquid from passing between the tube and the wall.

6. The high-pressure fluid apparatus according to claim 1, wherein the channel includes a rounded bend between 5° and 180°.

7. The high-pressure fluid apparatus according to claim 6, wherein the pipe follows the rounded bend.

8. The high-pressure fluid apparatus according to claim 1, wherein the pipe is made of a biocompatible material.

9. The high-pressure fluid apparatus according to claim 8, wherein the pipe is selected from stainless steel, titanium, stainless steel-nickel alloy, polyetheretherketone, and combinations thereof.

10. The high-pressure fluid apparatus according to claim 3, comprising a porous plug disposed adjacent to at least one of the inlet opening and the outlet opening, wherein the porous plug is located between the pipe and at least one of the inlet opening and the outlet opening, and the porous plug comprises one or more of stainless steel, titanium, stainless steel-nickel alloy, and polyetheretherketone.

11. An inlet configured to receive a liquid sample, It is a casing, A first component having a first contact surface and a channel extending through the first contact surface, A second component having a second contact surface, wherein the first component and the second component are fixed to each other by the first and second contact surfaces which are in a relationship of opposing relationships such that a contact portion is formed between them, and the second component and A housing including, A tube positioned within the channel of the first component, the tube defining a lumen having a substantially circular cross-section, A stationary phase medium disposed within the lumen of the tube, which is suitable for separating compounds present in the liquid sample by chromatography, A high-pressure fluid processing apparatus comprising an outlet port that is fluidly coupled to the inlet port via the aforementioned pipe.

12. The high-pressure fluid apparatus according to claim 11, wherein the depth of the channel is greater than or equal to the outer diameter of the pipe.

13. The high-pressure fluid apparatus according to claim 12, wherein the pipe is completely housed within the channel of the first component.

14. The high-pressure fluid apparatus according to claim 13, wherein the second contact surface is substantially planar, and the pipe does not intersect with the substantially planar second contact surface.

15. The high-pressure fluid apparatus according to claim 13, wherein the substantially circular cross-section of the pipe has a diameter of 1000 μm or less.

16. The high-pressure fluid apparatus according to claim 15, wherein the pipe includes its length, and the ratio of the length to the diameter is 100:1 or more.

17. The high-pressure fluid apparatus according to claim 11, wherein the gap between the channel and the pipe is filled with one or more epoxy resins, adhesives, and brazing materials.

18. To provide a first component having a first contact surface, The method involves forming a channel on the first contact surface of the first component, wherein the channel is formed to fluidly connect the inlet opening of the first component to the outlet opening of the first component. Positioning a tube containing a lumen having a substantially circular cross-section with a diameter of 1000 μm or less within the channel, The second component having a second contact surface is fixed to the first component, Positioning a filter at one of the aforementioned inlet openings and outlet openings, A method for forming a high-pressure fluid apparatus, comprising: filling the lumen with a stationary phase medium at a packing pressure of at least 5,000 psi, wherein the stationary phase medium is suitable for chromatographic separation of compounds present in a liquid sample, and the filter is impermeable to the stationary phase medium.

19. The method according to claim 18, comprising positioning a porous plug between the pipe and one of the inlet opening and the outlet opening.

20. The method according to claim 19, comprising positioning the porous plug before filling the lumen with the stationary phase medium.

21. The method according to claim 19, comprising positioning the porous plug after filling the lumen with the stationary phase medium.

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