The disclosed technology relates generally to a system for delivering a solvent having a temperature-independent solvent composition. More particularly, the disclosed technology relates to a system that reduces or eliminates the compositional error of the delivered solvent due to temperature-dependent viscosity and density changes of the solvents in a mobile phase of a liquid chromatography system.
Chromatography is a set of techniques for separating a mixture into its constituents. For instance, in a liquid chromatography application, a pump takes in and delivers one or more solvents, referred to as the mobile phase, to a sample manager where a sample is injected into the flow of the mobile phase. In an isocratic chromatography application, the composition of the mobile phase solvents remains unchanged, whereas in a gradient chromatography application, the solvent composition varies over time. The mobile phase, carrying the injected sample, passes through a column of particulate matter referred to as the stationary phase. By passing the mobile phase and sample through the column, the various components in the sample separate from each other at different rates and thus elute from the column at different times. A detector receives the elution from the column and produces an output from which the identity and quantity of analytes in the sample may be determined.
Solvent managers are used to generate and deliver the mobile phase to other components of the liquid chromatography system at precise flow rates, pressures, and solvent compositions. In some systems, solvents are metered and mixed at low pressure (i.e., ambient pressure). The metering can be achieved using a gradient proportioning valve which sequentially provides volume contributions of the different solvents used in the mobile phase. Generally, a solvent has a viscosity that is dependent on temperature. Similarly, the mass density of the solvent is dependent on temperature. A change in the viscosity of a solvent results in a different volume of the solvent contributed by the gradient proportioning valve even though the valve actuation time for the corresponding solvent remains constant. Likewise, a change in the density of a solvent results in a different mass of the solvent contributed by the gradient proportioning valve even if the contributed volume of the corresponding solvent were to be held constant. The relative changes in viscosity are different for the different solvents and the relative changes in density are different for the different solvents. Generally, changes in density have a significantly reduced effect on mass composition as compared to changes in viscosity.
Thus, changes in temperature generally cause a change in the solvent composition and the desired solvent composition may not be realized. Temperature changes may be due to variations in the ambient temperature, especially in environments that do not have accurate temperature control. Such temperature variations and the resulting changes in solvent composition affect the retention times of the analytes eluted from the column. Consequently, chromatography measurement data and data repeatability can be adversely affected by these temperature variations even though the volumes of the solvents may be accurately metered.
In one aspect, a liquid chromatography system includes a solvent manager and a temperature control module. The solvent manager includes a plurality of solvent reservoirs and a gradient proportioning valve in fluid communication with the plurality of solvent reservoirs. The temperature control module is disposed in the solvent manager and is configured to maintain a substantially constant temperature of at least one of the plurality of solvent reservoirs and the gradient proportioning valve.
The solvent manager may further include a degasser disposed between the plurality of solvent reservoirs and the gradient proportioning valve and the temperature control module may be configured to maintain a substantially constant temperature of at least one of the plurality of solvent reservoirs, the gradient proportioning valve, and the degasser. At least a portion of a fluidic path between the degasser and the gradient proportioning valve may be configured to maintain a temperature of the portion at a temperature of the gradient proportioning valve.
The temperature control module may include one or more temperature sensors and may include a thermal chamber. A heater may be disposed inside the thermal chamber and configured to heat an internal environment of the thermal chamber to a temperature greater than an ambient temperature. The thermal chamber may include an enclosure having a thermally controlled internal environment and substantially surrounding at least one of the plurality of solvent reservoirs and the gradient proportioning valve. The enclosure may include a thermally insulated housing.
The temperature control module may include an active thermal element in thermal communication with at least one of the plurality of solvent reservoirs and the gradient proportioning valve. The active thermal element may be a heater or a cooling device.
In another aspect, a solvent manager includes a plurality of solvent reservoirs, a gradient proportioning valve, a degasser, and a thermal chamber. The gradient proportioning valve is in fluid communication with the plurality of solvent reservoirs. The degasser is configured to receive and pass a flow of one or more solvents from the solvent reservoirs to the gradient proportioning valve. The thermal chamber has an enclosure with a thermally controlled internal environment and substantially surrounds at least one of the plurality of solvent reservoirs, gradient proportioning valve and degasser. The thermal chamber is configured to maintain a substantially constant temperature of at least one of the plurality of solvent reservoirs, gradient proportioning valve and degasser.
The solvent manager may further include an active thermal element disposed inside the thermal chamber. The active thermal element may be a heater or a cooling device.
At least one temperature sensor may be disposed inside the enclosure.
The above and further advantages of this invention may be better understood by referring to the following description in conjunction with the accompanying drawings, in which like numerals indicate like structural elements and features in the various figures. For clarity, not every element may be labeled in every figure. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating the principles of the invention.
Reference in the specification to “one embodiment” or “an embodiment” means that a particular, feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the teaching. References to a particular embodiment within the specification do not necessarily all refer to the same embodiment.
The present teaching will now be described in detail with reference to exemplary embodiments thereof as shown in the accompanying drawings. While the present teaching is described in conjunction with various embodiments and examples, it is not intended that the present teaching be limited to such embodiments and examples. On the contrary, the present teaching encompasses various alternatives, modifications, and equivalents, as will be appreciated by those of skill in the art. For example, various embodiments described herein refer to solvents although it should be recognized that other fluids can be used. Those of ordinary skill having access to the teaching herein will recognize additional implementations, modifications, and embodiments, as well as other fields of use, which are within the scope of the present disclosure as described herein.
Referring to
The illustrated system 10 includes a quaternary solvent manager (QSM) that meters and mixes up to four solvents at low (i.e., ambient) pressure. The solvent mixture is pressurized to a system pressure level by the pump system 34 before delivery to the downstream components of the system 10. The GPV 26 includes a plurality of fluid switching valves (e.g., solenoid valves) that are in fluid communication, by tubing or other form of fluidic conduit, with respective solvent component reservoirs 50A, 50B, 50C and 50D. A degasser 46 disposed between the solvent reservoirs 50 and the GPV 26 removes dissolved gases from the individual solvents. The GPV outlet port is coupled to the inlet port of the pump system 34. The solvent mixture is delivered from the pump outlet port to a chromatographic column 54, typically at a substantially higher pressure than the pressure of the solvent mixture exiting the GPV 26.
During operation of the liquid chromatography system 10, the switching valves of the GPV 26 are opened sequentially for each metering cycle so that the pump system 34 draws a volume of fluid from each of the reservoirs 50 contributing to the solvent mixture. The volume proportions of solvents present in the solvent mixture depend on the actuation times for each of the switching valves in relation to the inlet velocity profile during the intake cycle. Thus, the mass composition of the fluid mixture is also determined by the actuation times.
The viscosity and density of a solvent are dependent on the solvent temperature. Moreover, the viscosity change of a solvent for a given temperature change may be different for different solvents. Similarly, the change in the density of a solvent for a given temperature change can differ between the solvents. Generally, if the temperature of the solvents change, the composition of the solvent mixture delivered by the pump system 34 also changes. Thus, ambient temperature changes may cause the solvent mixture composition to differ from the desired solvent composition. A consequence of a temperature change can be a significant increase or decrease in retention times.
An experimental setup was configured to evaluate the effect of temperature variations on the performance of two ACQUITY ARC® liquid chromatography systems. Each system had similar components, including a quaternary solvent manager QSM-R, a flow through needle FTN-R sample manager, a CM-A column heater and a 2489 ultraviolet/visible detector. In addition, each system used a CORTECS® 4.6×50 mm, 2.7 μm C18 BEH column. These systems and components are available from Waters Corporation of Milford, Mass.
The sample used to determine retention time was alkylphenone in a 90:10 water:acetonitrile diluent. The gradient composition was a water and acetonitrile mixture that transitioned linearly from 10% acetonitrile to 60% acetonitrile over 15 minutes and the flow rate was 0.8 mL/minute.
To affect temperature changes on each liquid chromatography system in a controlled manner, the system was placed in a temperature-controlled chamber and the chamber was maintained at different temperatures between approximately 4° C. to approximately 40° C. while the room temperature was maintained at 22° C.±0.5° C. Injections were performed at different temperatures across the temperature range to determine the dependence of retention time on temperature.
To better determine the liquid chromatography system components that are more thermally sensitive with respect to retention time, an evaluation was performed using different configurations of a solvent manager of the liquid chromatography systems. Each configuration included an arrangement in which one or more solvent manager components were temperature-controlled while the remainder of the solvent manager components and the other system components were exposed to a range of temperatures, again between about 4° C. to about 40° C. To achieve this arrangement in an experimental setup, injections were performed with the liquid chromatography system disposed inside a temperature-controlled chamber while one or more components were “temperature controlled,” that is, maintained at ambient temperature by locating the component(s) outside the chamber and coupling the solvent flow between the outside component(s) and the chamber through tubing. In this manner, components outside the chamber were not subject to the range of internal temperatures generated by the chamber.
It can be seen from
In embodiments of a liquid chromatography system having reduced sensitivity to temperature variations, temperature control of the gradient proportioning valve, solvent reservoirs and/or degasser may be accomplished in a variety of ways. For example, a temperature control module may be disposed inside the solvent manager and used to regulate the temperature of one or more of these system components. In one example, the temperature control module may include a thermal chamber configured to maintain a substantially constant temperature of one or more of the components. As used herein, a substantially constant temperature means a temperature that varies only by an amount that does not lead to any measurable change in retention time or at least a change in retention time that does not change the characteristics of a chromatogram as perceived by one of skill in the art. The thermal chamber may include an enclosure that substantially surrounds all components that are temperature regulated and provides a thermally controlled internal environment. The enclosure may include a thermally insulated housing to decrease thermal conductance between the regulated components and the environment external to the housing. Optionally, multiple thermal chambers may be used if more than one system component is temperature controlled. In a different example, the gradient proportioning valve, solvent reservoirs and/or degasser may be thermally controlled via an active thermal element (e.g., a heater) in direct thermal communication with the component through a thermally conductive path. In some implementations the heater is used to maintain a temperature that is above the ambient temperature for the liquid chromatography system.
In some of the examples discussed above, the temperature control module is described as having a heater to establish the desired temperature of the regulated system components although it should be recognized that a cooling device could instead be used. Generally, a heater is preferred due to lower cost. The temperature of the controlled system components should be set at a safe margin above the highest ambient temperature anticipated for the instrument environment. For example, the regulated temperature may be 5° C. or 10° C. above the nominal ambient temperature. Conversely, if regulation is achieved by cooling, the regulated temperature should be 5° C. or 10° C. less than the nominal ambient temperature. The regulated temperature is preferably maintained at a substantially constant temperature. For example, the temperature may be considered to be substantially constant if the minimum and maximum values of the regulated temperature over time differ by no more than 1° C.
The temperature control module may include one or more temperature sensors disposed near or at the solvent reservoirs 50, degasser 46 and/or GPV 26. For example, the temperature sensors may be used as part of a control loop for thermal control. For example, the temperature sensors can be thermocouples or thermistors and may be mounted to measure the air temperature within an enclosure that surrounds the temperature-controlled components.
Regulating the temperature of one or more of the GPV 26 and the other components of the low pressure mixing portion of the chromatography system results in a reduction in retention time thermal dependence. Regulating all the low-pressure components, that is, the GPV 26, solvent reservoirs 50 and degasser 46, so that they are at a substantially constant temperature results in substantially no retention time thermal dependence. In this regard, substantially no thermal dependence of retention time means the retention time may vary but such variations would not render any detrimental effect on retention time data for chromatographic peaks as interpreted by one of skill in the art. Thus, users would not consider the variations in retention time to render the system measurement data unsuitable for typical chromatographic separations.
While the invention has been shown and described with reference to specific embodiments, it should be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention as recited in the accompanying claims.
This application claims the benefit of the earlier filing date of U.S. Provisional Patent Application Ser. No. 63/145,833, filed Feb. 4, 2021, and titled “Thermally-Controlled Low Pressure Mixing System for Liquid Chromatography,” the entirety of which is incorporated herein by reference.
Number | Date | Country | |
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63145833 | Feb 2021 | US |