Single unit device for viscosity and light scattering and method thereof

Abstract

A single cell device for viscosity and light scattering and methods thereof are disclosed. The single unit device may include an inlet line, first and second fluid flow lines in fluid communication with the inlet line, a light scattering detector (LSD), a pressure transducer line, a pressure transducer disposed in fluid coupling with the pressure transducer line, and a discharge stream. The first fluid flow line may include first and second capillary tubes and a first three-way connector interposed between the first and second capillary tubes. The second fluid flow line may include first and second capillary tubes and a second three-way connector interposed between the first and second capillary tubes. The LSD may be disposed downstream of the second three-way connector and upstream of the second capillary of the second fluid flow line. The discharge flow may be in fluid communication with respective second capillaries of the first fluid flow line and the second fluid flow line.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 358,348, filed on July 5, 2022, the contents of which are incorporated herein by reference to the extent consistent with the present disclosure. Background Art

[0003] In liquid chromatography, such as in size exclusion chromatography, the experimental throughput is usually determined or limited by the rate of sample preparation and separation rather than the detection time. Therefore, multiple detectors are conventionally placed after sample preparation and separation process to efficiently and simultaneously obtain more property information from a single chromatographic run. Common detectors for measuring concentration and chemical identification can include detectors for refractive index, ultraviolet absorption, infrared absorption, etc. Common detectors for measuring macromolecular microstructures can include viscosity and light scattering detectors, which usually require supporting concentration detectors to refer to each chromatographic data point. These detectors can be arranged in series or in parallel with each other. Since the jet of liquid chromatography remains below the laminar flow, the corresponding signal of each continuous detector is widened according to the pipeline or flow path (for example, pipeline) or the detector arranged upstream thereof. It is also necessary to adjust the relative signal time for the jet delay between detectors. In order to solve the phenomenon of signal broadening and jet delay between detectors, it is usually necessary (for example, via an algorithm) to utilize a method for mathematical correction.

[0004] While these mathematical corrections are well established in the field of liquid chromatography, it is also well known that these mathematical corrections result in reduced chromatographic resolution. For example, mathematical corrections can often include convolution and offset of upstream detectors to match the most widespread and subsequent or latest eluting detectors, which results in reduced chromatographic resolution. In contrast, avoiding detector deconvolution in the field of liquid chromatography is not encouraged or taught due to the amplification of noise during conventional deconvolution.

[0005] In addition to the foregoing, it should be understood that the detectors are arranged in series or in a series configuration with the disadvantage of causing the expansion of additive detectors due to downstream or subsequent detectors. The use of a series configuration may also result in difficulty in generating excessive back pressure on an upstream detector. In addition, some detectors, such as a viscometer, may dilute the sample concentration, and some detectors, such as a conventional refractometer, may have a relatively large waste liquid pipeline, which may aggravate the widening. The detectors are arranged in parallel or in a parallel configuration with the disadvantage of additional widening over time due to the reduced flow rate in the parallel configuration. In addition, due to defects in one of the pipelines (e.g., scaling), the flow through the parallel pipelines may change over time. The parallel configuration also typically requires the corresponding waste liquid pipelines of each detector to be combined to avoid gravity effects.

[0006] Therefore, improved viscosity and light scattering detectors are needed to solve the above technical problems. Summary of the invention

[0007] The present invention content is only intended to introduce a simplified overview of some aspects of one or more embodiments of the present disclosure. Based on the specific embodiments provided below, other application areas of the present disclosure will become apparent. The present invention content is not an exhaustive overview, nor is it intended to determine the key or important elements of this teaching, nor is it intended to delimit the scope of the present disclosure. On the contrary, its purpose is only to present one or more concepts in a simplified form as a preface to the specific embodiments below.

[0008] The foregoing and / or other aspects and utilities embodied in the present disclosure can be achieved by providing a single unit device. The single unit device may include: an inlet pipeline, a first fluid flow pipeline, a second fluid flow pipeline, a light scattering detector, a pressure transducer pipeline, a pressure transducer and / or an exhaust stream. The first fluid flow pipeline may include a first capillary in direct fluid communication with the inlet pipeline; a second capillary arranged in series with the first capillary; and a first three-way connector inserted between the first capillary and the second capillary of the first fluid flow line. The second fluid flow may be in fluid communication with the inlet. The second fluid flow line may include a first capillary in direct fluid communication with the inlet pipeline; a second capillary arranged downstream of the first capillary; and a second three-way connector inserted between the first and second capillaries of the second fluid flow line. The light scattering detector may be arranged downstream of the second three-way connector and upstream of the second capillary of the second fluid flow line. The pressure transducer line may fluidically couple the first three-way connector with the second three-way connector. The pressure transducer may be arranged and fluidly coupled to the pressure transducer pipeline. The exhaust stream may be in fluid communication with the second capillary of the first fluid flow line and the second capillary of the second fluid flow line.

[0009] In at least one embodiment, the single unit device may further include a dilution reservoir disposed downstream of the light scattering detector and upstream of the second capillary of the second fluid flow line.

[0010] In at least one embodiment, the light scattering detector may include a sample cell, which may include: an inlet coupled to the first capillary fluid of the second fluid flow line and disposed downstream of the first capillary; and first and second outlets coupled to the second capillary fluid of the second fluid flow line and disposed upstream of the second capillary.

[0011] In at least one embodiment, the sample pool may also include: a body defining a flow path extending axially therethrough, the flow path may include a cylindrical inner section interposed between a first outer section and a second outer section, wherein the first outer section is a truncated cone, and a first end of the first outer section is directly fluidically connected to the inner section and has a cross-sectional area relatively smaller than a cross-sectional area at its second end, wherein the body also defines an inlet of the inner section that is directly fluidically connected to the inner section and is configured to direct a sample to the flow path, and wherein the body also defines a first and a second outlet, wherein the first outlet and the second outlet are configured to couple the first and second outer sections to an exhaust flow via a second capillary fluid of a second fluid flow line.

[0012] In at least one embodiment, the second outer section of the sample cell may be frustoconical, and the first end of the second outer section may be in direct fluid communication with the inner section and may have a relatively smaller cross-sectional area than the second end thereof.

[0013] In at least one embodiment, the body can define a first recess extending axially therethrough, and the first recess can be in fluid communication with the first outer section and configured to receive a first lens of the light scatter detector.

[0014] In at least one embodiment, the body can define a second recess extending axially therethrough, and the second recess can be in fluid communication with the second outer section and configured to receive a second lens of the light scatter detector.

[0015] In at least one embodiment, the body of the sample cell can define a bore extending radially therethrough, wherein the bore can be in direct fluid communication with the inner section of the flow path.

[0016] In at least one embodiment, the single unit device may further include an optically transparent material disposed in the hole.

[0017] In at least one embodiment, the single unit device can also include one or more purge lines fluidly coupled to the pressure transducer and configured to purge the pressure transducer.

[0018] In at least one embodiment, the single unit device may further include a respective purge valve disposed in each of the one or more purge lines, each of which may optionally be fluidly coupled to the exhaust stream.

[0019] In at least one embodiment, the light scattering detector may further include a laser for emitting a light beam, wherein the flow path of the sample cell may have a centerline aligned with the light beam.

[0020] In at least one embodiment, the light scattering detector may further include at least one detector operably coupled to the sample cell and configured to receive scattered light emitted from the sample cell.

[0021] The foregoing and / or other aspects and utilities embodied in the present disclosure may be achieved by providing a system comprising: a single unit device disclosed herein or according to any of the preceding paragraphs, and a refractometer operably coupled to the single unit device.

[0022] In at least one embodiment, the single unit device and the refractometer may be operably coupled in series with each other.

[0023] In at least one embodiment, the refractometer may be positioned upstream of the single unit device.

[0024] The foregoing and / or other aspects and utilities embodied in the present disclosure can be achieved by providing a method of using any of the systems disclosed herein, which can include the single unit device and a refractometer operably coupled to the single unit device. The method can include: flowing a sample through the refractometer; and flowing a sample through the single unit device.

[0025] In at least one embodiment, flowing the sample through the single unit device may include flowing the sample from an inlet line via a first fluid flow line and a second fluid flow line to an outlet stream.

[0026] In at least one embodiment, flowing the sample through the second fluid flow line may include flowing the sample through a first capillary and a second capillary of the second fluid flow line, and flowing the sample through a light scattering detector interposed between the first capillary and the second capillary of the second fluid flow line.

[0027] In at least one embodiment, the method may further include flowing the sample from the refractometer to the single unit device. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The accompanying drawings, which are incorporated into and constitute a part of this specification, illustrate different embodiments of the present disclosure. These and / or other aspects and advantages in the embodiments of the present disclosure will become apparent and more easily understood from the following description of the embodiments in conjunction with the accompanying drawings. It should be noted that some details of the accompanying drawings have been simplified and drawn to facilitate understanding of the present disclosure rather than to maintain strict structural accuracy, details and proportions. These drawings / diagrams are intended to be illustrative and not restrictive.

[0029] FIG. 1 shows a schematic diagram of a conventional viscometer according to the prior art.

[0030] FIG. 2A shows a schematic diagram of a conventional sample cell for a light scattering detector according to the prior art.

[0031] 2B shows an enlarged view of a portion of a sample cell indicated by the box labeled 2B of FIG. 2A according to the prior art.

[0032] FIG. 2C shows a schematic diagram of a light scattering detector (LSD) including a sample cell according to the prior art.

[0033] Figure 3 A schematic diagram of an exemplary combination or single unit device incorporating a viscometer and a light scattering detector is shown according to one or more disclosed embodiments.

[0034] Figure 4 A schematic diagram of another exemplary single unit device incorporating a viscometer and a light scattering detector is shown in accordance with one or more disclosed embodiments.

[0035] Figure 5 A graph of the observed light scattering angle and viscometer signal for Example 1 is shown.

[0036] Figure 6 A schematic diagram of a comparative single unit device incorporating a viscometer and a light scattering detector is shown.

[0037] Figure 7 Shows Figure 6 A plot comparing the observed signals of a right angle light scattering (RALS) detector and a viscometer for a single unit device.

[0038] Figure 8 Shows Figure 4 Plot of the observed signals of the right angle light scattering (RALS) detector and viscometer of a single unit device.

[0039] Fig. 9 Shown is a schematic diagram of a viscometer and a light scattering detector in a parallel configuration.

[0040] Fig.10 A schematic diagram of a viscometer and a light scattering detector in series or in a tandem configuration is shown.

[0041] Fig.11 A schematic diagram of a viscometer and light scattering detector in an exemplary single unit device configuration is shown.

[0042] Fig.12 Shown in Fig. 9 Color peak diagram observed in parallel configuration.

[0043] Fig.13 Shown in Fig.10 Color peak diagram observed in tandem configuration.

[0044] Fig.14 Shows the use of Fig.11 Color peak diagram observed for a single unit device configuration. DETAILED DESCRIPTION

[0045] The following description of various exemplary aspects is merely exemplary in nature and is in no way intended to limit the present disclosure, its application, or uses.

[0046] As used throughout this disclosure, ranges are used as shorthand for describing each value within the range. It should be recognized and understood that the description in range format is only for convenience and brevity, and should not be interpreted as a hard limit to the scope of any embodiment or implementation disclosed herein. Therefore, the disclosed range should be interpreted as having specifically disclosed all possible sub-ranges and individual numerical values ​​within the range. Therefore, any value within the range can be selected as the endpoint of the range. For example, the description of a range such as from 1 to 5 should be considered to have clearly disclosed sub-ranges, such as 1.5 to 3, 1 to 4.5, 2 to 5, 3.1 to 5, etc.; and individual numbers within the range, for example, 1, 2, 3, 3.2, 4, 5, etc. This applies regardless of how wide the range is.

[0047] In addition, all numerical values ​​are "about" or "approximately" indicated values, and experimental errors and variations that would be expected by a person of ordinary skill in the art are taken into account. It should be understood that all numerical values ​​and ranges disclosed herein are approximate values ​​and ranges, whether or not "about" is used in conjunction with it. It should also be understood that the term "about" as used herein in conjunction with a number refers to ±0.01% (inclusive), ±0.1% (inclusive), ±0.5% (inclusive), ±1% (inclusive), ±2% (inclusive) of the number, ±3% (inclusive) of the number, ±5% (inclusive) of the number, ±10% (inclusive) of the number, or ±15% (inclusive) of the number. It should also be understood that when a numerical range is disclosed herein, any numerical value falling within the range is also specifically disclosed.

[0048] All references cited herein are incorporated by reference in their entirety. In the event of a conflict in a definition in the present disclosure and that of a cited reference, the present disclosure controls.

[0049] Fig. 1 shows a schematic diagram of a conventional viscometer 100 according to the prior art. The conventional viscometer 100 shown in Fig. 1 may also be referred to as a Wheatstone bridge viscometer 100, which is further described in detail in the following documents: Haney, MA (1985). The Differential Viscometer. IA New Approach to the Measurement of Specific Viscosities of Polymer Solutions. Journal of Applied Polymer Science, Vol. 30, 3023-3036., Haney, MA (1985). The Differential Viscometer. II. On-Line Viscosity Detector for Size-Exclusion Chromatography. Journal of Applied Polymer Science, Vol. 30, 3037-3049, and U.S. Pat. No. 4,463,598 issued on Aug. 7, 1984, the contents of which are incorporated herein in the context of the present disclosure.

[0050] As shown in FIG1 , a conventional viscometer 100 may include an inlet 101 fluidly coupled to a plurality of capillaries R1-R4 (four 102, 104, 106, 108 are shown). In at least one embodiment, the capillaries 102, 106 may form a first fluid flow line 105, and the capillaries 104, 108 may form a second fluid flow line 103. The first fluid flow line 105 may include a "tee" connection 113 interposed between the capillaries 102, 106. The second fluid flow line 103 may include a "tee" connection 112 interposed between the capillaries 104, 108.

[0051] Any one or more of the capillaries 102, 104, 106, 108 may have a resistance that is equally matched to any one or more of the remaining capillaries 102, 104, 106, 108 or may have a resistance that is known not to match it. The conventional viscometer 100 of FIG. 1 may include a delay reservoir or delay column 110 disposed downstream of the "tee" connection 112. The delay reservoir 110 may be capable of or configured to delay the sample flowing through the viscometer 100 from entering the reference capillary 108, thereby causing a measured resistance mismatch across or via a bridge or pressure transducer line 111 and / or a pressure transducer coupled to the line 111. It should be understood that the discharge stream or outlet line 114 of the viscometer 100 may be diluted by about 50% concentration. It will also be appreciated that a sample broadening effect is observed in conventional viscometers 100 due to the length of the capillaries 102, 104, 106, 108 and the split flow involved, which will generally broaden the elution flow exiting the detector via the exhaust flow 114. For the foregoing reasons, when the conventional viscometer 100 shown in FIG. 1 is used in series or in a serial configuration, it is typically configured toward or at the end of the serial configuration to avoid dilution and sample broadening effects.

[0052] FIG. 2A shows a schematic diagram of a conventional sample pool 200 for a light scattering detector 242 according to the prior art. FIG. 2B shows an enlarged view of a portion of the sample pool 200 indicated by the box marked 2B of FIG. 2A according to the prior art. The conventional sample pool 200 and the light scattering detector 242 are further described in detail in the following document: Haney, Max. "Light Scattering Detectors and Sample Cells for the Same." Patent Cooperation Treaty (PCT) PCT / US2019 / 012090, filed on January 2, 2019, the contents of which are incorporated herein by reference within the scope consistent with the present disclosure. As shown in FIGS. 2A and 2B, the sample pool 200 may include a body 202 that defines a flow path 204 extending axially therethrough. The flow path 204 may define the volume of the sample pool 200. The flow path 204 may include a central axis or centerline 282 extending therethrough and configured to define the overall orientation of the flow path 204. As shown in FIG. 2B , the flow path 204 and its central axis 282 can be aligned or coaxial with the light beam 280 emitted from the laser 256 of the light scattering detector 242. The flow path 204 can include a cylindrical inner section 206 interposed between a first outer section 208 and a second outer section 210. The first outer section 208 can be frustoconical, and the first end 212 of the first outer section 208 can be in direct fluid communication with the inner section 206 and can have a cross-sectional area that is relatively smaller than the cross-sectional area at its second end 214. The body 202 can also define an inlet 216 that is in direct fluid communication with the inner section 206 and is configured to direct the sample to the inner section 206 of the flow path 204, such as the middle of the inner section 206. The second outer section 210 can be frustoconical, and the first end 218 of the second outer section 210 can be in direct fluid communication with the inner section 206 and can have a cross-sectional area that is relatively smaller than the cross-sectional area at its second end 220.

[0053] The body 202 may also define a first outlet 222 and a second outlet 224 extending therethrough. The first outlet 222 and the second outlet 224 may be configured to fluidly couple the respective second ends 214, 220 of the first outer section 208 and the second outer section 210 to a waste or outlet line 213 via a first outlet line 226 and a second outlet line 228, respectively. As shown in FIG. 2A , the body 202 may define a first recess 230 extending axially therethrough. The first recess 230 may be in fluid communication with the first outer section 208 and configured to receive a first lens 232 of a light scattering detector 242. The body 202 may define a second recess 234 extending axially therethrough. The second recess 234 may be in fluid communication with the second outer section 210 and configured to receive a second lens 236 of the light scattering detector 242. As shown in FIG. 2B , the body 202 may define a hole 238 extending radially therethrough. The well 238 can be in direct fluid communication with the inner section 206 of the flow path 204, such as a middle portion of the inner section 206. The sample cell 200 can also include an optically transparent material 240 disposed in the well 238.

[0054] Typically, the sample cell 200 shown in Fig. 2A and Fig. 2B can include a single inlet 216 and two outlets 226, 228. The two outlets can have matching flow or flow velocity. The sample cell 200 in Fig. 2A and Fig. 2B at least maximizes sensitivity while minimizing the instant band broadening at the measuring point. It should be understood that the conventional sample cell 200 shown in Fig. 2A and Fig. 2B is not ideally used in a series configuration. For example, sample broadening may occur in the waste liquid line or the outlet line 213 leaving the sample cell 200. In view of the above, when the light scattering detector 242 including the conventional sample cell 200 shown in Fig. 2A and Fig. 2B is used in series, it is usually arranged toward the end of the series configuration or at its end.

[0055] Fig. 2C shows a schematic diagram of an exemplary light scattering detector (LSD) 242 including the sample pool 200 shown in Fig. 2A and Fig. 2B according to the prior art. It should be understood that both static and dynamic light scattering detectors are considered. LSD 242 can be operably coupled with a sample source or device 244, and can or be configured to receive a sample or effluent therefrom. For example, as shown in Fig. 2C, LSD 242 can be coupled with a sample source or device 244 fluid via pipeline 246 and be configured to receive effluent therefrom. Exemplary sample source or device 244 can include but is not limited to a chromatographic instrument that can or be configured to separate one or more analytes of a sample or eluent from each other. For example, a sample source or device 244 can be a liquid chromatography instrument, as known in the art, which can or be configured to separate them from each other based on the charge (e.g., ion exchange chromatography) of the analyte of the eluent, size (e.g., size exclusion or gel permeation chromatography), etc. In an exemplary embodiment, LSD 242 is operably coupled to a liquid chromatography instrument that is configured to separate analytes from one another based on their respective sizes. For example, LSD 242 is operably coupled to a liquid chromatography instrument that includes a gel permeation chromatography column.

[0056] The LSD 242 may include an exemplary sample cell 200, a collimated light beam such as a laser 256, and one or more detectors 258, 260, 262 (three are shown) operably coupled to each other. The detectors 258, 260, 262 may be any suitable detector capable of or configured to receive analyte scattered light. For example, any one or more of the detectors 258, 260, 262 may be a photodetector, such as a silicon photodetector. The LSD 242 may include one or more lenses 232, 236, 264, 266, 268 (five are shown) capable of or configured to refract, focus, attenuate and / or collect light transmitted through the LSD 242, and one or more mirrors 270, 272 (two are shown) capable of or configured to reflect or redirect light transmitted through the LSD 242.

[0057] In at least one embodiment, the first lens 232 and the second lens 236 can be disposed on opposite sides of the sample cell 200 and configured to refract, focus, attenuate and / or collect light transmitted therethrough. In another embodiment shown in FIGS. 2A and 2C , the body 202 of the sample cell 200 can define a first recess 230 and a second recess 234 extending longitudinally or axially therethrough and configured to receive the first lens 232 and the second lens 236, respectively. As shown in FIGS. 2A and 2C , each of the first and second lenses 232, 236 can define a convex surface along its respective first or outer end 248, 250. Although the first ends 248, 250 of the first and second lenses 232, 236 are shown as defining a convex surface, it should be understood that any of the respective first ends 248, 250 of the first and second lenses 232, 236 can alternatively define a flat surface. As further shown in FIG. 2A , each of the first and second lenses 232, 236 can define a flat surface along its respective second or inner end 252, 254. The respective second ends 252 , 254 of the first and second lenses 232 , 236 may seal and / or at least partially define a flow path 204 extending through the sample cell 200 .

[0058] The laser 256 can be any suitable laser capable of or configured to provide a light beam 280 having a sufficient wavelength and / or power. For example, the laser 256 can be a diode laser, a solid-state laser, etc. The laser 256 can be configured to emit a light beam 280 that passes through the sample cell 200. For example, as shown in FIG2C , the laser 256 can be arranged or disposed around the LSD 242 so that the light beam 280 emitted therefrom is transmitted through the sample cell 200. As further shown in FIG2C , a third lens 264 can be interposed between the sample cell 200 and the laser 256 and configured to focus the light beam 280 directed to the sample cell 200 and passing through it.

[0059] In at least one embodiment, at least one of the reflectors 270, 272 can be associated with a respective detector 258, 260 and configured to reflect or redirect light (e.g., scattered light or analyte scattered light) toward the respective detector 258, 260. For example, as shown in FIG. 2C , a first reflector 270 can be disposed near the first lens 232 and configured to reflect at least a portion of the light from the first lens 232 toward the first detector 258. In another example, a second reflector 272 can be disposed near the second lens 236 and / or interposed between the second lens 236 and the third lens 264 and configured to reflect at least a portion of the light from the second lens 236 toward the second detector 260. In at least one embodiment, one or more lenses 266, 268 can be interposed between the first and second reflectors 270, 272 and the first and second detectors 258, 260 to focus, refract, or otherwise direct light from the reflectors 270, 272 to the detectors 258, 260. For example, as shown in FIG. 2C , the fourth lens 266 may be interposed between the first detector 258 and the first reflector 270 , and the fifth lens 268 may be interposed between the second detector 260 and the second reflector 272 .

[0060] In at least one embodiment, at least one of the detectors 258, 260, 262 can be configured to receive light (e.g., scattered light or analyte scattered light) from the sample pool 200 without the aid or reflection of one of the mirrors 270, 272. For example, as shown in FIGS. 2B and 2C, the third detector 262 can be disposed adjacent to or coupled to the sample pool 200 and configured to receive light (e.g., scattered light) from the sample pool 200 at an angle of approximately 90° relative to the light beam 280. As further discussed herein, the optically transparent material or sixth lens 240 can be configured to refract or direct the scattered light toward the third detector 262.

[0061] As shown in FIG. 2C , at least one of the sample pool 200, the first, second and third lenses 232, 236, 264 and the first and second reflectors 270, 272 can be arranged parallel to each other, coaxially or otherwise aligned along the direction of the light beam 280 emitted by the laser 256. As further shown in FIG. 2C , each of the first and second detectors 258, 260 can be arranged or positioned to receive light (e.g., scattered light or analyte scattered light) from the corresponding reflector 270, 272 in a direction substantially perpendicular to the light beam 280 emitted by the laser 256. Each of the first reflector 270 and the second reflector 272 can define a corresponding hole or path 274, 276 extending therethrough. For example, the first reflector 270 can define a hole 274 extending therethrough in a direction parallel, coaxial or otherwise aligned with the light beam 280. Similarly, the second reflector 272 can define a hole 276 extending therethrough in the direction parallel, coaxial or otherwise aligned with the light beam 280. It should be appreciated that the holes 274 , 276 extending through the respective mirrors 270 , 272 may allow the light beam 280 emitted from the laser 256 to be transmitted through the first and second mirrors 270 , 272 , thereby preventing the light beam 280 from being reflected toward the first and second detectors 258 , 260 .

[0062] In at least one embodiment, the LSD 242 can include one or more screens or apertures 284, 286. For example, as shown in FIG2C, a first aperture can be interposed between the first lens 232 and the first reflector 270, and a second aperture can be interposed between the second reflector 272 and the lens 264. The apertures 284, 286 can be configured to "clean up", isolate, or otherwise filter stray light (e.g., halos) from the light beam 280. For example, the apertures 284, 286 can define a hole or aperture (e.g., an adjustable aperture / diaphragm) that can or is configured to filter out stray light from the light beam 280.

[0063] It should be understood that a viscometer such as conventional viscometer 100 shown in FIG. 1 and a light scattering detector such as light scattering detector 242 in FIG. 2C are often used in conjunction with refractometer 900 ( Fig. 9 and Fig.10 For example, viscometer 100 and light scattering detector 242 are often fluidically and / or operatively coupled to refractometer 900. Conventional methods of coupling viscometer 100 and light scattering detector 242 to refractometer 900 often require a 3-way parallel splitter (e.g., Fig. 9 as shown) or in series configuration (as Fig.10As shown in FIG. 1 ). As is known in the art, a 3-way parallel splitter is relatively difficult to maintain and introduces technical problems that increase the spread of all three detectors (e.g., viscometer, light scattering detector, and refractometer). In addition, as is known in the art, a series configuration of three detectors will dilute or spread the downstream or subsequent detectors.

[0064] In view of the foregoing, the inventors surprisingly and unexpectedly discovered that combining the viscometer 100 and the light scattering detector 242 into a combination or single unit 300, 400 or "unit" solves the aforementioned technical problems. Specifically, the inventors surprisingly and unexpectedly discovered that combining the viscometer 100 and the light scattering detector 242 into a single unit 300, 400 eliminates the need or requirement for a 3-way parallel splitter ( Fig. 9 ). The inventors have also surprisingly and unexpectedly discovered that when the viscometer 100 and the light scattering detector 242 are combined into a single unit 300, 400 as disclosed herein, the delay volumes and peak shapes of the viscometer 100 and the light scattering detector 242 substantially match or exhibit a relatively closer match than conventional methods utilizing a serial or parallel configuration. The combined or single unit 300, 400 combining the viscometer 100 and the light scattering detector 242 can be easily used in a serial configuration with another detector, such as a UV detector (not shown). For example, the single unit 300, 400 can be used downstream of the UV detector in a serial configuration. The single unit 300, 400 combining the viscometer 100 and the light scattering detector 242 can also be easily used in a parallel configuration with another detector, such as a refractometer, to determine concentration.

[0065] Figure 3 A schematic diagram of an exemplary single unit device 300 incorporating a viscometer 100, a light scattering detector 242, and a sample cell 200 thereof is shown in accordance with one or more disclosed embodiments. The viscometer 100, the light scattering detector 242, and the sample cell 200 thereof may be similar in some respects to the viscometer 100, the light scattering detector 242, and the sample cell 200 described above; therefore, they may be best understood with reference to the description of FIG. 1 and FIG. 2A to FIG. 2C, where like reference numbers indicate like components and will not be described in detail again. Figure 3 As shown, the light scattering detector 242 is disposed downstream or after the "tee" connection 112, which couples the sample flowing through the capillary 104 to the pressure transducer 302 and the capillary 108. The "tee" connection 112 may also be referred to as a "tee" connector or joint. Exemplary "tee" connectors may be or include, but are not limited to, joints, tees, and crosses for high pressure HPLC connections, which may be obtained from Thermo Fisher Scientific TM(Catalog numbers: 03-052-437, 03-052-438, 03-170-306, etc.) are commercially available. Figure 3 As further shown, the light scattering detector 242 is disposed upstream of the delay column or dilution reservoir 110. For simplicity, Figure 3 The schematic diagram of FIG. 242 also uses a sample cell 200 to represent the LSD 242 and its sample cell 200. Surprisingly and unexpectedly, it was found that Figure 3 The light scattering detector 242, and in particular the specific location of its sample cell 200, is shown to significantly affect the observed match width of the viscometer 100. The surprising and unexpected results are demonstrated in the following example.

[0066] Figure 4 A schematic diagram of another exemplary single unit device 400 incorporating a viscometer 100, a light scattering detector 242, and a sample cell 200 thereof is shown in accordance with one or more disclosed embodiments. The viscometer 100, the light scattering detector 242, and the sample cell 200 thereof may be similar in some respects to the viscometer 100, the light scattering detector 242, and the sample cell 200 described above; therefore, they may be best understood with reference to the description of FIG. 1 and FIG. 2A to FIG. 2C , where like reference numbers indicate like components and will not be described in detail again. Figure 4 As shown, the single unit device 400 may include one or more wash lines (two wash lines 402, 404 are shown), each of the wash lines 402, 404 having at least one wash valve 406, 408 coupled thereto. The wash lines 402, 404 and the wash valves 406, 408 coupled thereto may be capable of or configured to wash the pressure transducer 302 of the viscometer 100 without introducing any stagnant solvent into the sample cell 200 of the light scattering detector 242. In addition, the wash lines 402, 404 and the wash valves 406, 408 coupled thereto allow liquid or sample to bypass the light scattering detector 242 and its sample cell 200, thereby providing a means for changing solvents or adjusting columns. The wash lines 402, 404 and the wash valves 406, 408 coupled thereto may also allow for changes in solvents and / or adjustments to the column while keeping the light scattering detector 242 free of air, fouling materials, and particles. Although Figure 4 Rinse lines 402, 404 are shown coupled to outlet 114, but it should be understood that rinse lines 402, 404 may be fluidly coupled to any waste or outlet line capable or configured to rinse transducer 302. Rinse lines 402, 404 need not be in fluid communication with common outlet 114.

[0067] The single unit devices 300, 400 described herein combine the viscometer 100, the light scattering detector 242 and / or the sample cell 20 thereof and are Figure 3 and Figure 4 It is shown in the figure, providing one or more of the following technical effects:

[0068] (1) Minimizing peak broadening and shift between the viscometer 100 and the light scattering detector 242 compared to conventional series and parallel configurations of the viscometer and light scattering detector;

[0069] (2) minimizing conventional mathematical corrections, which are known to degrade resolution or introduce extraneous noise or artifacts, especially when multi-detector dosing is required;

[0070] (3) resulting in minimal or relatively small shape (e.g., peak) differences compared to conventional means and methods;

[0071] (4) resulting in minimal or relatively small offset between the viscometer 100 and the light scattering detector 242 as compared to conventional configurations or methods utilizing viscometers and light scattering detectors (e.g., series or parallel configurations);

[0072] (5) minimizing sample dilution in the viscometer 100 or light scattering detector 242;

[0073] (6) minimizing the possibility or opportunity of introducing certain substances into the light scattering detector 242, including when the viscometer detector 100 is cleaning its transducer 302;

[0074] (7) Maintaining a constant or substantially constant back pressure on the light scattering detector 242 to minimize solvent degassing from its sample cell 200 while not relying on adding more external tubing to other detectors for split flow balance or back pressure as in a parallel or parallel configuration.

[0075] With respect to technical effect (3), it will be understood by those skilled in the art that the "shape" of a chromatographic peak is defined as or refers to the peak width at half height (50% height). Further with respect to the technical effect of (3), i.e., resulting in minimal or relatively small shape differences compared to conventional methods, such as compared to standard and tandem configurations, it is noted that this effect is more pronounced or more easily observed when the sample is a monodisperse standard injected into a liquid chromatograph.

[0076] Although Figure 3 and Figure 4A corresponding single unit device 300, 400 utilizing a combination of an exemplary viscometer 100 and an exemplary light scattering detector 242 is shown, but it should be understood that the light scattering detector 242 may be replaced and / or supplemented with any other suitable detector. For example, the light scattering detector 242 may be replaced and / or supplemented with any other suitable detector capable of or configured to provide any one or more of the technical effects disclosed herein. Contemplated exemplary detectors may be or include, but are not limited to, a refractive index (RI) detector, an ultraviolet (UV) detector, an infrared (IR) detector, a fluorescence detector, a conductivity detector, the like, or a combination thereof.

[0077] The following numbered paragraphs disclose one or more exemplary variations of the subject matter of the present application:

[0078] 1. A single unit device, comprising: an inlet pipeline; a first fluid flow pipeline in fluid communication with the inlet pipeline, the first fluid flow pipeline comprising: a first capillary in direct fluid communication with the inlet pipeline; a second capillary arranged in series with the first capillary; and a first three-way connector inserted between the first capillary and the second capillary of the first fluid flow line; a second fluid flow line in fluid communication with the inlet pipeline, the second fluid flow line comprising: a first capillary in direct fluid communication with the inlet pipeline; a second capillary arranged downstream of the first capillary; a second three-way connector inserted between the first capillary and the second capillary of the second flow line; a light scattering detector arranged downstream of the second three-way connector and upstream of the second capillary of the second fluid flow line; a pressure transducer pipeline fluidically coupling the first three-way connector and the second three-way connector; a pressure transducer arranged in the pressure transducer pipeline; an exhaust stream in fluid communication with the second capillary of the first fluid flow line and the second capillary of the second fluid flow line.

[0079] 2. The single unit device of paragraph 1, further comprising a dilution reservoir disposed downstream of the light scattering detector and upstream of the second capillary of the second fluid flow line.

[0080] 3. A single unit device according to paragraph 1 or 2, wherein the light scattering detector includes a sample cell, the sample cell comprising: an inlet coupled to the first capillary fluid of the second fluid flow line and arranged downstream of the first capillary; and a first and second outlet coupled to the second capillary fluid of the second fluid flow line and arranged upstream of the second capillary.

[0081] 4. A single unit device according to paragraph 3, wherein the sample pool further comprises: a body defining a flow path extending axially therethrough, the flow path comprising a cylindrical inner section interposed between a first outer section and a second outer section, wherein the first outer section is truncated conical and a first end of the first outer section is directly fluidically connected to the inner section and has a cross-sectional area relatively smaller than a cross-sectional area at a second end thereof, wherein the body further defines an inlet of the inner section that is directly fluidically connected to the inner section and is configured to direct a sample to the flow path, and wherein the body further defines the first and second outlets, wherein the first outlet and the second outlet are configured to couple the first and second outer sections to the discharge flow via the second capillary fluid of the second fluid flow line.

[0082] 5. A single unit device according to paragraph 4, wherein the second outer section of the sample pool is frustoconical, and the first end of the second outer section is in direct fluid communication with the inner section and has a cross-sectional area that is relatively smaller than the cross-sectional area at its second end.

[0083] 6. A single unit device according to paragraph 4 or 5, wherein the body defines a first recess extending axially therethrough, the first recess being in fluid communication with the first outer segment and being configured to receive a first lens of the light scattering detector.

[0084] 7. A single unit device according to paragraph 6, wherein the body defines a second recessed portion extending axially therethrough, the second recessed portion being in fluid communication with the second outer section and configured to receive a second lens of the light scatter detector.

[0085] 8. The single unit device of any of paragraphs 4 to 7, wherein the body of the sample cell defines an aperture extending radially therethrough, wherein the aperture is in direct fluid communication with the inner section of the flow path.

[0086] 9. The single unit device of paragraph 8, further comprising an optically transparent material disposed in the aperture.

[0087] 10. The single unit device of any of paragraphs 1 to 9, further comprising one or more cleaning lines fluidly coupled to the pressure transducer and configured to clean the pressure transducer.

[0088] 11. The single unit device of paragraph 10, further comprising a respective purge valve disposed in each of the one or more purge lines, optionally each of the one or more purge lines being fluidly coupled to the exhaust stream.

[0089] 12. A single unit device according to any of paragraphs 4 to 11, wherein the light scattering detector further comprises a laser for emitting a light beam, wherein the flow path of the sample cell has a centerline aligned with the light beam.

[0090] 13. The single unit device of paragraph 12, wherein the light scattering detector further comprises at least one detector operably coupled to the sample cell and configured to receive scattered light emitted from the sample cell.

[0091] 14. A system comprising: a single unit device according to any of the preceding paragraphs; and a refractometer operably coupled to the single unit device.

[0092] 15. The system of paragraph 15, wherein the single unit device and the refractometer are operably coupled in series with each other.

[0093] 16. The system of paragraphs 14 or 15, wherein the refractometer is positioned upstream of the single unit device.

[0094] 17. A method of using the system of any of paragraphs 14 to 16, the method comprising: flowing a sample through the refractometer; and flowing the sample through a single unit device.

[0095] 18. The method of paragraph 17, wherein flowing the sample through the single unit device comprises flowing the sample from the inlet line to the outlet stream via the first fluid flow line and the second fluid flow line.

[0096] 19. A method according to paragraph 18, wherein causing the sample to flow through the second fluid flow line includes causing the sample to flow through the first capillary and the second capillary of the second fluid flow line, and causing the sample to flow through the light scattering detector inserted between the first capillary and the second capillary of the second fluid flow line.

[0097] 20. The method of any of paragraphs 17 to 19, further comprising flowing the sample from the refractometer to the single unit device.

[0098] Examples

[0099] The examples and other embodiments described herein are exemplary and are not intended to be limiting in describing the full scope of the compositions and methods of the present disclosure. Equivalent changes, modifications and variations can be made to specific embodiments, materials, compositions and methods within the scope of the present disclosure, and have substantially similar results.

[0100] Example 1

[0101] Described in detail above and by Figure 4 The single unit device 400 shown is used to measure three light scattering angles and viscometer signals. Specifically, low angle light scattering (LALS), right angle light scattering (RALS) and high angle light scattering (HALS) are measured at about 10 °, about 90 ° and about 170 °, respectively. The sample used is a narrow polystyrene standard with a nominal molecular weight of about 96,100 Da, which can be purchased from Tosoh Bioscience, LLC. (Tosoh Bioscience, PA). The sample uses tetrahydrofuran (THF) as a solvent at a concentration of about 1.05 mg / mL. The conditions for operating the single unit device 400 are as follows: an injection volume of about 100 μL, a flow rate of about 1 mL / min, and a chromatographic column GMHHR-H is used, which can be purchased from Tosoh Bioscience, LLC. Figure 5 The measurement diagram is shown in Figure 2. Figure 5 As shown, all three observed light scattering angles and observed viscometer signals exhibit simultaneous or substantially simultaneous measurements having the same or substantially the same shape. It should be understood that Figure 5 The observed light scattering angles and viscometer signals shown in are raw signals. In other words, the observed light scattering angles and viscometer signals are raw signals that have not been smoothed, deconvolved, shifted, or otherwise manipulated.

[0102] Example 2

[0103] It should be understood that one of ordinary skill in the art can adequately numerically define the chromatographic peak shape of a monodisperse component by describing the peak width at half height (50% height). It should also be understood that one of ordinary skill in the art can adequately measure tailing by measuring the peak width at 1 / 5 height (20% height).

[0104] A comparative single-unit device having a different structure from the single-unit device 400 of Example 1 was evaluated. Specifically, Figure 6 A comparative single unit device 600 incorporating a viscometer 100 and a light scattering detector 242 having configuration A was tested and compared to the exemplary single unit device 400 of Example 1 and was shown in FIG. Figure 4 Shown in. Figure 6 The viscometer 100, light scattering detector 242 and sample cell 200 of the present invention may be similar to the viscometer 100, light scattering detector 242 and sample cell 200 described above in some aspects; therefore, reference may be made to FIGS. 1, 2A to 2C, Figure 3 and Figure 4They are best understood by referring to the following description, in which like reference numerals refer to like parts and will not be described in detail again. Figure 6 As shown, the light scattering detector 242 and its sample cell 200 are placed after or downstream of the capillary 104 and before or upstream of the T-fitting 112. The viscometer detector 100 and the right angle light scattering (RALS) detector 262 are monitored simultaneously and the baseline is subtracted. Figure 7 and Figure 8 The results of the comparative single unit device 600 and the exemplary single unit device 400 of Example 1 are shown respectively. The results of the comparative single unit device 600 and the exemplary single unit device 400 of Example 1 are summarized in the following Table 1 ( Figure 4 ) The corresponding differences or differences (Δ) in peak width at 50% (e.g., shape), 20% (e.g., tailing), and the difference in peak retention volume.

[0105] Table 1

[0106]

[0107] As shown in Table 1 and Figure 7 and Figure 8 As shown, numerically and visually, the exemplary single unit device 400 ( Figure 4 ) in the peak elution shape and the synchronization of the peak time, thereby facilitating the placement of the light scattering detector 242 and its sample cell 200 in the Figure 3 and Figure 4 Configuration shown.

[0108] Example 3

[0109] To compare the series / serial, parallel, and combined configurations, a refractometer was placed in series before the parallel and series configurations to serve as a reference detector. The comparison was made using the same light scattering detector 242 and viscometer 100, which were configured as follows: (1) a parallel detector (e.g. Fig. 9 (2) a series detector, with the viscometer 100 in front (as shown); Fig.10 and (3) utilizing a "combined" configuration of an exemplary single unit device 400 of the present disclosure (such as Fig.11 The results are summarized in Table 2 below. Fig.12 , Fig.13 and Fig.14 The corresponding graphs of the observed / measured color peaks in the parallel configuration, the series configuration, and the combined configuration are shown, respectively.

[0110] Table 2

[0111]

[0112] It should be noted that the refractometer detector cell volume and outlet tubing is approximately 150 μL, which accounts for the majority of the observed delay volume shown in Table 2. It is observed that the “combined” configuration (e.g. Fig.14 The average detector shape in the combined configuration (shown in Figure 4) is significantly, surprisingly and unexpectedly improved. The average hold-up volume shift is similar to the series configuration, but with the advantage that the RALS area is not reduced. The combined configuration is more advantageous than the parallel configuration, which has approximately half the restored viscometer area due to the reduced parallel flow. Therefore, using the "combined configuration" or exemplary single unit device 400 provides the maximum signal and restores the best or closest peak shape relative to the refractive index detector.

[0113] The present disclosure has been described with reference to exemplary embodiments. Although a limited number of embodiments have been shown and described, those skilled in the art will appreciate that changes may be made in these embodiments without departing from the principles and spirit of the foregoing specific embodiments. The present disclosure is intended to be interpreted as including all such modifications and variations as long as they fall within the scope of the appended claims or their equivalents.

Claims

1. A single unit device comprising: Inlet pipeline; a first fluid flow line in fluid communication with the inlet line, the first fluid flow line comprising: a first capillary tube in direct fluid communication with the inlet line; a second capillary tube arranged in series with the first capillary tube; and a first three-way connector interposed between the first capillary tube and the second capillary tube of the first fluid flow line; a second fluid flow line in fluid communication with the inlet line, the second fluid flow line comprising: a first capillary tube in direct fluid communication with the inlet line; a second capillary tube disposed downstream of the first capillary tube; a second three-way connector interposed between the first capillary tube and the second capillary tube of the second flow line; a light scattering detector disposed downstream of the second three-way connector and upstream of the second capillary of the second fluid flow line; a pressure transducer line fluidly coupling the first three-way connector and the second three-way connector; a pressure transducer disposed in the pressure transducer pipeline; An exhaust stream is in fluid communication with the second capillary of the first fluid flow line and the second capillary of the second fluid flow line.

2. The single unit device of claim 1, further comprising a dilution reservoir disposed downstream of the light scattering detector and upstream of the second capillary of the second fluid flow line.

3. A single unit device according to claim 1 or 2, wherein: The light scattering detector comprises a sample cell, and the sample cell comprises: an inlet fluidly coupled to the first capillary of the second fluid flow line and disposed downstream of the first capillary; and First and second outlets are fluidly coupled to the second capillary tube of the second fluid flow line and disposed upstream of the second capillary tube.

4. A single unit device according to claim 3, wherein: The sample pool also includes: a body defining a flow path extending axially therethrough, the flow path including a cylindrical inner section interposed between a first outer section and a second outer section, wherein the first outer section is truncated-conical, and a first end of the first outer section is in direct fluid communication with the inner section and has a cross-sectional area that is relatively smaller than a cross-sectional area at a second end thereof, wherein the body further defines an inlet in direct fluid communication with the inner section and configured to direct a sample to the inner section of the flow path, and Wherein the body further defines the first and second outlets, wherein the first outlet and the second outlet are configured to fluidly couple the first and second outer segments to the exhaust stream via the second capillary of the second fluid flow line.

5. A single unit device according to claim 4, wherein: The second outer section of the sample cell is frustoconical, and a first end of the second outer section is in direct fluid communication with the inner section and has a cross-sectional area that is relatively smaller than a cross-sectional area at a second end thereof.

6. A single unit device according to claim 4 or 5, wherein: The body defines a first recess extending axially therethrough, the first recess being in fluid communication with the first outer section and configured to receive a first lens of the light scatter detector.

7. A single unit device according to claim 6, wherein: The body defines a second recess extending axially therethrough, the second recess being in fluid communication with the second outer section and configured to receive a second lens of the light scatter detector.

8. A single unit device according to any one of claims 4 to 7, wherein: The body of the sample cell defines a bore extending radially therethrough, wherein the bore is in direct fluid communication with the inner section of the flow path.

9. The single unit device of claim 8, further comprising an optically transparent material disposed in the aperture.

10. The single unit device of any preceding claim, further comprising one or more purge lines fluidly coupled to the pressure transducer and configured to purge the pressure transducer.

11. The single unit device of claim 10, further comprising a respective purge valve disposed in each of the one or more purge lines, optionally each of the one or more purge lines being fluidly coupled to the exhaust stream.

12. A single unit device according to any one of claims 4 to 11, wherein: The light scattering detector further comprises a laser for emitting a light beam, wherein the flow path of the sample cell has a centerline aligned with the light beam.

13. The single unit device of claim 12, wherein: The light scattering detector also includes at least one detector operably coupled to the sample cell and configured to receive scattered light emitted from the sample cell.

14. A system comprising: A single unit device according to any one of the preceding claims; as well as A refractometer is operably coupled to the single unit device.

15. The system according to claim 15, wherein: The single unit device and the refractometer are operably coupled in series with each other.

16. The system according to claim 14 or 15, wherein: The refractometer is disposed upstream of the single unit device.

17. A method of using the system according to any one of claims 14 to 16, the method comprising: flowing a sample through the refractometer; as well as The sample is flowed through the single unit device.

18. The method according to claim 17, wherein: Flowing the sample through the single unit device includes flowing the sample from the inlet line to the exhaust stream via the first fluid flow line and the second fluid flow line.

19. The method according to claim 18, wherein: Flowing the sample through the second fluid flow line includes flowing the sample through the first capillary and the second capillary of the second fluid flow line, and flowing the sample through the light scattering detector interposed between the first capillary and the second capillary of the second fluid flow line.

20. The method of any one of claims 17 to 19, further comprising flowing the sample from the refractometer to the single unit device.

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