EP3679368A1

EP3679368A1 - Method for operating a fractionation collector during chromatography

Info

Publication number: EP3679368A1
Application number: EP18765095.7A
Authority: EP
Inventors: Alexander BOZIC
Original assignee: Individual
Current assignee: Individual
Priority date: 2017-09-05
Filing date: 2018-09-03
Publication date: 2020-07-15
Also published as: US11402359B2; DE102017120421A1; US20200284769A1; WO2019048369A1

Abstract

The present invention relates to a method for operating a fractionation collector during chromatography, wherein a solvent composition is pumped into a chromatography column which contains at least one portion of a first solvent which is liquid under normal conditions and a portion of a second solvent which is gaseous under normal conditions, and at least part of the composition flowing out of the chromatography column is introduced into a detector, wherein a composition containing the first solvent, after leaving the chromatography column, is introduced into a collection container of the fraction collector in accordance with the detector signal, and the amount of liquid which is introduced into a collection container of the fraction collector is selected in accordance with the portion of the first solvent. The present invention also relates to a chromatography system for carrying out the present method.

Description

Method for operating a fractionation collector in a

chromatography

The present invention relates to a method for operating a

Fractionation collector in a chromatography as well as a chromatography unit for carrying out the method.

Many advantages can be achieved by supercritical fluid chromatography (SFC)

Substances can be separated particularly easily and reliably, chemically analyzed, identified and quantified. When using carbon dioxide (C0 ₂ ) as a liquid in SFC applications, the extraction of the substances is generally carried out above the critical temperature of 31 ° C and above a critical pressure of 74 bar.

To C0 ₂ or a C0 ₂ mixture in a liquid state within a

Keep Chromatography column, the entire chromatography system must be kept at a predetermined pressure level. For this purpose is

downstream of the chromatographic column and downstream of each

Detector typically provided a back pressure regulator to maintain the pressure within the chromatography system at a predetermined level.

In practical applications, the SFC technology suffers from the disadvantage that the mobile phase of the chromatographically separated substances can not easily be collected in opened vessels. Once a mixture of liquid C0 ₂ and an additional solvent is exposed to atmospheric pressure, C0 ₂ expands and forms an aerosol with the additional solvent.

In an SFC, however, the composition of the aerosol can vary widely, as many times a solvent gradient is used to separate substances. The mixture of C0 ₂ and an additional solvent, such as methanol, may vary, for example, from 10% to 60% methanol fraction. As a result, the constitution of the aerosol and its volume flow can vary accordingly, resulting in suboptimal separation rates of gaseous and liquid fractions of the aerosol

Aerosols in a cyclone separator leads. Other gas-liquid separation systems use, for example, an impingement deposition, wherein the volume flow of the aerosol is directed to a baffle plate, which may optionally be provided by a test tube. In general, impingement separators and inertial separators require one

comparatively large volume into which the aerosol can expand. Such comparatively large vessels are not optimal in terms of self-cleaning effects, since cross-contamination of aerosols and substances may occur which are sequentially processed by such separators. In particular, the transit time differences of the substances must be very large to one

ensure adequate separation. Basically, the size and the surface of the baffle separator can be minimized when operating at elevated pressure level.

For example, a serving as a baffle test tube may be provided in a pressurized environment. The aerosol may then escape from a bent outlet and may impact the side wall of the test tube at a predetermined angle. With such a shock absorber, it is indeed possible to collect smaller quantities of a substance at a much lower cost. However, shock absorbers operating at an elevated pressure level do not allow large scale automated fractionation to be realized.

Thus, the operating costs and installation costs are relatively high, since only a limited amount of test tubes in the printing area can be processed automatically. In addition, the separation rate is not as good as with settlers operating at atmospheric pressure.

When using gas-liquid separators, such as those in

WO 2014/012962 A1, this problem can be solved. However, the separation of large amounts of gas leads to a change in the volume flow originally introduced into the chromatography column. This can cause problems with fraction collection. For example, in the paper by Rui Chen, "Mass-Directed Preparative SFC with Open-Bed Fraction Collection," Waters Corporation, May 2010, 72000346en states that signal integrity may be compromised as a solution to this problem at room temperature liquid Add solvent before relaxing by a pump to the

To keep volume flow constant.

However, this solution is very time-consuming and error-prone. It should also be noted that any change of a collection vessel during the collection of a fraction results in a relatively high loss. The switching or changing time is about 1 second, during which a volume to be collected is directed into a waste container. At a flow rate of, for example, 150 ml / min there is a loss of 2.5 ml. Larger reservoirs could reduce this problem. However, these are also associated with disadvantages, as this requires a larger area. Furthermore, the losses due to adherence to the vessel surface are relatively high for small volumes. However, multiple rinsing of the collection vessels requires the processing of unnecessarily diluted compositions.

In view of the prior art, it is therefore an object of the present invention to provide a method for operating a fractionation collector in a chromatography, which solves the problems outlined above. Here, the method should be as simple and inexpensive to carry out and lead to further cost and handling advantages. For example, signal integrity should be achieved without the addition of solvent. Furthermore, the degree of filling of the collecting container, even with a change of the

Partialvolumenstroms of a liquid solvent under normal conditions be as high as possible.

Furthermore, the provision of a method for operating a

Fractionation Collector Object of this invention, in which the highest possible yield of the purified substances can be achieved. In this case, this high yield should be achievable for as different gas-liquid mixtures as possible.

Another object is by a chromatography system for

To provide a method that can be operated and manufactured particularly cost and low maintenance. Furthermore, an object of the present invention provide components that allow the simplest possible conversion of a known HPLC system to an SFC system. Here also a conversion of preparative HPLC plants should be made possible.

These and other tasks which are not explicitly mentioned, but which can be readily deduced or deduced from the contexts discussed hereinbelow, are solved by a method for operating a system

Fractionation collector in a chromatography with all features of claim 1.

The present invention accordingly provides a method for operating a fractionation collector in a chromatography, wherein a solvent composition is pumped into a chromatography column containing at least a proportion of a first solvent, the

Normal conditions is liquid, and contains a proportion of a second solvent which is gaseous under normal conditions, and the composition flowing from the chromatography column is at least partially introduced into a detector, which is characterized in that a composition containing the first solvent after leaving the Chromatography column is introduced in response to the detector signal in a collecting vessel of the fraction collector and the amount of liquid which is introduced into a collecting vessel of the fraction collector, depending on the proportion of the first solvent is selected.

The present invention in particular causes the

Profitability is improved, in particular, a high degree of filling of the collecting vessels of the fraction collector can be achieved. Furthermore, the

Signal integrity are maintained without the need for a metered addition of solvent.

Furthermore, the process for operating a fractionation collector can be carried out very inexpensively and simply. In addition, a very achieved high yield of the purified substances. This high yield is achieved for many different gas-liquid mixtures.

Furthermore, very good results are also achieved in chromotography processes using gradient gas-liquid compositions. In addition, the procedure can be very different

Volume flows of the aerosol are carried out without the economic benefits would be severely impaired.

Furthermore, an automated fractionation is possible, which can be scaled according to the needs, without the need for large investments.

Further, the present method as well as the chromatographic equipment for carrying out a process can reduce the complexity and cost of the technical equipment required for the establishment of the SFC analysis. Also, HPLC equipment may be reconstructed for a preparative process Insert are provided.

In a chromatography process according to the invention is a

Solvent composition is pumped into a chromatography column containing at least a proportion of a first solvent, which at

Normal conditions is liquid, and contains a proportion of a second solvent which is gaseous under normal conditions. It is therefore preferably an SFC, as set forth above and below.

Normal conditions mean 273.15 K = 0 ° C and 1, 01325 bar according to DIN 1343.

To perform a separation with a supercritical fluid is a

inorganic or organic solvent used, which is liquid under the usual separation conditions, preferably at 25 ° C and atmospheric pressure (1023mbar). In this case, a polar or nonpolar solvent can be used, depending on the nature of the compounds to be separated or purified. These

In the present case, substances are referred to in particular as the first solvent.

Preferably, the first solvent is selected from alcohol, preferably

Methanol, ethanol or propanol, hexane, mixtures with dichloromethane, Chloroform, water (preferably up to a maximum of 3% by volume, otherwise a miscibility gap may occur), an aldehyde or a ketone, preferably

methyl ethyl ketone; an ester, preferably ethyl acetate; or an ether, preferably tetrahydrofuran, an aliphatic hydrocarbon, preferably hexane, cyclohexane, heptane, octane; an aromatic hydrocarbon,

preferably benzene, toluene, xylene. These compounds can be used singly or as a mixture.

Furthermore, it can be provided that in a method according to the invention preferably a gas is used, which can be relatively easily put into a supercritical state. Among the preferred gases having these properties include carbon dioxide (CO ₂ ), ammonia (NH ₃ ), freon, xenon, with carbon dioxide (CO ₂ ) being particularly preferred. These substances are referred to herein in particular as a second solvent.

It can preferably be provided that the gas-liquid mixture to be brought into the supercritical state comprises a polar solvent and a gas which is selected from the group consisting of C0 ₂ , NH ₃ , freon, xenon, preferably C0 ₂ . Preferably, the polar solvent is an alcohol, preferably methanol, ethanol or propanol, hexane, mixtures with

Dichloromethane, chloroform, water (preferably up to a maximum of 3% by volume, otherwise a miscibility gap may occur), an aldehyde or a ketone, preferably methyl ethyl ketone; an ester, preferably ethyl acetate; or an ether, preferably tetrahydrofuran.

It can further be provided that the gas-liquid mixture to be brought into the supercritical state comprises a non-polar solvent and a gas which is selected from the group consisting of C0 ₂ , NH ₃ , freon, xenon,

preferably C0 ₂ . Preferably, the non-polar solvent is an aliphatic hydrocarbon, preferably hexane, cyclohexane, heptane, octane; on

aromatic hydrocarbon, preferably benzene, toluene, xylene; an ester, preferably ethyl acetate; or an ether, preferably tetrahydrofuran.

The composition flowing from the chromatography column is at least partially introduced into a detector according to the present method.

Accordingly, preferably a check of the corresponding Composition made after the chromatography column. Detectors which can be used for this purpose are well known, in particular spectroscopic methods in which electromagnetic waves are used, such as UV, VIS spectroscopy are performed. In this case, it is also possible to use further detection methods which measure, for example, light scattering, fluorescence or the refractive index. Furthermore, mass spectrometers and / or conductivity detectors, etc. are often used.

These methods can measure, continuously or batchwise, the properties of the composition flowing out of the chromatography column so that these detectors can determine these properties in flow or by sampling, the latter being generally fully automated and

continuously. Details of these techniques are known in the art, including, in particular, detectors used in conventional HPLC methods.

According to the invention, a composition containing the first solvent is introduced after leaving the chromatography column in dependence on the detector signal in a collecting vessel of the fraction collector and the amount of

Liquid, which is introduced into a collecting vessel of the fraction collector, depending on the proportion of the first solvent.

By this measure can be achieved in particular that the volume of a collecting vessel of the fraction collector can be exploited in a surprisingly good manner. As a result, in particular the cost and handling advantages set out above can be achieved.

Further cost and handling benefits can be achieved by not introducing at least a portion of the composition containing the first solvent into a collection vessel after leaving the chromatography column. Preferably, compositions that do not contain valuable substances are discarded, which is generally controlled by a control valve in the

Fraction collector is effected, the shares to be discarded the first

Solvent-containing composition after leaving the

Chromatography column into a waste container or similar directs. In a particular embodiment it can be provided that the solvent composition pumped into a chromatography column is changed in the course of the chromatography.

Preferably, the proportion of the first solvent in the

Increased solvent composition in the course of chromatography and reduced the proportion of second solvent in the course of chromatography, with particular preference, the proportion of the first solvent at the beginning of

Chromatography at least 10 vol .-%, more preferably at least 20 vol .-% below the proportion of the first solvent at the end of the chromatography, based on the solvent composition. Accordingly, the proportion of solvent which is liquid under normal conditions preferably increases, while the proportion of solvent which is gaseous under normal conditions decreases. By this configuration can surprisingly encrustation in one

Gas separator, which is used in a preferred embodiment of the method minimized, preferably completely prevented. As a result, the separation quality of the system or the process can be surprisingly improved.

Furthermore, it can be provided that the proportion of first solvent in the range of 5 to 95 vol .-% and the proportion of the second solvent in the range of 5 to 95 vol .-%, based on the solvent composition.

It can preferably be provided that the chromatography is carried out at a pressure in the range of 50 to 500 bar, preferably 75 to 400 bar.

In a preferred embodiment it can be provided that the

Chromatography at a temperature in the range of 20 ° C to 80 ° C, preferably 35 ° C to 60 ° C is performed.

In a preferred embodiment it can be provided that the second solvent, which is contained in the composition after the chromatography column, is at least partially separated before being introduced into the fraction collector. As a result, a significant improvement in the economy can be achieved surprisingly, since a part of the solvent before the

Fraction collector separated and thereby at the same volume of Collection vessels rarely a change of collection vessels must be made.

Accordingly, it can be provided that a gas-liquid separator is used, wherein the pressure in the gas-liquid separator preferably in the range of 0.1 to 4 bar, preferably 0.5 to 3.5 bar and particularly preferably 1 to 3 bar is located (overpressure). By means of this embodiment, further surprising improvements can be achieved. In particular, the running time between relaxing the

Solvent mixture and the outlet of the gas-liquid separator in

Essentially determined by the pressure drop. The resulting gas in the small volume causes a high pressure, which determines the duration. The relative constancy of this pressure leads to a high signal integrity, since the running time remains substantially constant even after relaxing with the use of a suitable gas-liquid separator. Accordingly, especially when using a gas-liquid separator, which can be operated preferably at an overpressure, dispensed with a metered addition of solvent, as was used in the prior art, to ensure signal integrity. It should be noted that an absolute

Constancy of the gas pressure in the gas-liquid separator is not necessary, since the runtime is very short even at relatively low pressures, so that fluctuations have no significant effect on the signal integrity, as this is subject to a usual fluctuation due to fault tolerances. To lead

Pressure fluctuations in the range of 0.1 bar to 4 bar in the gas-liquid separator to appropriate results. Preferred embodiments of suitable gas-liquid separators will be described later, to which reference is made.

In a preferred embodiment of the method in which the chromatographic system comprises a back pressure regulator, by means of which the pressure in the gas-liquid separator is controllable, it can be provided that the regulation of the pressure is selected as a function of the solvent content of the gas-liquid mixture , Preferably, the control can be designed so that at a high

Solvent content is provided a high pressure in the gas-liquid separator. Preferably, it may be provided that the fractionation is operated at a lower pressure than the gas-liquid separator, wherein preferably the pressure difference in the range of 0.1 to 4, preferably 0.5 to 3.5 bar and particularly preferably 1 to 3 bar is located.

The fractionation is preferably carried out at a pressure in the range from 0 to 1 bar (overpressure), more preferably 0 to 0.5 bar, especially preferably 0 to 0.2 bar. The pressure values set out above refer to overpressure, which pressure is measured relative to atmospheric pressure or air pressure.

The detection of a fraction to be collected can be determined in a conventional manner, which are generally also used in related chromatography methods. This includes, for example, that a fraction at a certain signal level of the detector, for example, a UV / VIS detector is collected. Furthermore, a fraction can also be collected on the basis of a specific form of the signal, for example the predetermined change in the slope of the detector signal or a specific value of the slope of the signal

Detector signal.

Preferably, it may be provided that in determining the time of the collection start for the initiation of the fractions to be collected the

Fluid volume of the lines and devices between the detector to the separation point is taken into account, in which the fractions to be collected is separated from the fractions to be discarded. This period of time through which a

Calibration of the system is effected, thus enabling an optimal collection of the valuable fractions, can be carried out by methods known per se. For example, a trial run can be carried out on the basis of which

Time span can be estimated. In a preferred embodiment, for example, a dye can be used, which is visible on the fraction collector. The time span between the detector signal, for example a UV / VIS signal, and the appearance of the colored fraction on the fraction collector gives these

Period of time. In principle, this period can of course also be determined via the measured volume and the set volume flow (flow rate). These methods are known from the prior art since these Problem with any HPLC system with an automatic fractionation exists and is solved.

In a preferred embodiment, it may be provided that the proportion of the first solvent in the solvent composition is determined by the time at which the fraction collection begins, with a control unit preferably providing the time interval over the course of time

Solvent composition is entered and based on the

Start time of the collection, the proportion of the first solvent is determined and based on this proportion and the flow rate, the collection time for a

Collection is set, wherein in the control unit, the volume of the collecting vessel is deposited.

Accordingly, the partial volume of the first solvent is preferably determined at the time of detection, this value being determined, for example, from the

Pumping capacity of the pump with the first solvent in the

Chromatography column is transported, the gradient of this pump power and the time of detection can be obtained. From these values, the partial volume of the first solvent can be determined at the time of detection, which can preferably be used to determine the appropriate utilization of the volume of the collecting vessel. The real one

Collection of the detected fraction is preferably made taking into account the amount of time it takes for the liquid to be collected to travel the distance between the detector signal and the fraction collection valve.

Furthermore, it can be provided that the degree of filling of the collecting vessel takes place via a summation of the volume, which is introduced into the collecting vessel over a period of time, wherein the individual volume components of the summation on the products of the flow rate, the period over which the

Volume component is collected, and the proportion of the first solvent in the solvent composition at the start time of the respective

Volume component is determined, wherein in the control unit, the volume of the collecting vessel is deposited.

This relationship can be represented, for example, by the following formula (I):

t = t _s

Formula (I) where:

F _G is the total volume flow φ _(t) is the volume fraction of the first solvent at time t

At is the period of time over which the volume of a single volume component is calculated

V _max is the maximum volume of the receiver t _s is the start time at which the detector signal indicates a fraction to be collected t _n is the end time to which a fraction to be collected enters

Collection vessel can be initiated

Accordingly, the total volume is continuously calculated from the sum of the individual volume components and at the time (t _n ) at which this is done

Total volume equal to the maximum volume of the collection vessel, sent to the fraction collector a signal to carry out a change of the collecting vessel. Here, the maximum volume of the collecting vessel (V _max ) is chosen so that overflow of the collecting vessel can be avoided.

Furthermore, it can be provided that the guided into a collecting vessel

Liquid amount is at least 70%, preferably at least 90% of the maximum volume for the collecting volume of the collecting vessel, if the detection unit does not detect a new fraction. As a result, a very good utilization of the provided volume of the collection vessels can be achieved. Upon detection of a new fraction, the collection vessel is generally changed. Preferably, it can be provided that the period over which the volume of a single volume component is calculated, in the range of 0.01 to 10.0 s, preferably 0.1 to 5.0 s and more preferably 0.5 to 2.0 s is.

Furthermore, it can be provided that the chromatography at a flow rate in the range of 10 ml / min to 450 ml / min, more preferably in the range of 50 ml / min to 300 ml / min and especially preferably 100 ml / min to 250 ml / min is carried out. This flow rate represents the total flow rate. The flow rate of the individual solvents, in particular of the first and the second

Solvent, which are each used as a mixture, results from the respective volume fraction.

In a preferred embodiment, the detector comprises a

Mass spectrometers and the time required to determine the mass spectrometry signal is taken into account in the control unit of the fraction collector. Preferably, when using a mass spectrometer, an additional volume may be provided, which is arranged between the mass spectrometer and the fraction collector, so that the flow time of the fraction to be collected is large enough to allow reliable detection.

Furthermore, it can be provided that the fraction collector via a

Controlling unit is controlled and the control unit is in operative connection with the detector, wherein upon detection of a substance by the detector, a control pulse is passed to the fraction collector, a change of the

Collecting vessel causes.

In a further embodiment it can be provided that the

Fraction collector is controlled by a control unit and the

Control unit is in operative connection with the detector, wherein after

Termination of the detection of a substance by the detector a control pulse is passed to the fraction collector, which causes a change of the collecting vessel. This embodiment is preferred over the embodiment in which a change of the collecting vessel is effected at the beginning.

Another object of the present invention is a chromatography plant for performing a method, wherein the chromatography plant a Controlling, which is in operative connection with a detector and a fraction collector and the controller is programmable so that the amount of liquid that can be introduced into a vessel of the fraction collector, depending on the proportion of the first solvent is fixable.

With regard to preferred embodiments of a plant for carrying out a method according to the invention, reference is made to the above-described embodiments, which were set out with regard to the method according to the invention, in order to avoid repetitions.

A Chromatography Plant, which is for a supercritical

Liquid chromatography is designed, for example, using supercritical C0 ₂ together with a solvent such as methanol. Accordingly, a chromatography unit designed for supercritical fluid chromatography has at least one storage tank for the first solvent and a storage tank for the supercritical fluid, for example C0 ₂ . In general, the fluid is out of the store

taken and transferred with a respective at least one pump into which a mixing element which is in flow communication with a chromatography column. The pumps and / or the mixing element and the chromatography column can be provided with a temperature control in order to be able to set a given temperature in each case. For this purpose, in particular, heat exchangers can be provided. The addition of mixtures to be separated, in particular to be purified

Substances can be carried out by known devices, for example injectors, which are preferably provided in the conduit in which the solvent is passed to the mixing element.

It can preferably be provided that the chromatography system a

Injection device comprises, with which automatically samples in the

Chromatographieanlage can be injected.

The fluid leaving the chromatographic column is preferably at least partially supplied to a detection or analysis unit. Examples of one

Detection or analysis unit include UV detectors and / or

Mass spectrometry. After the chromatography column and preferably after the detection or analysis unit is generally a back pressure regulator and preferably after the back pressure regulator, a heat exchanger is provided. The aerosol leaving the heat exchanger is preferably subsequently fed to a gas-liquid separator.

Preferably to be used gas-liquid separator are known from the prior art, for example, the document WO 2014/012962 A1, wherein the

Revelation of this document is incorporated by reference in the present application for disclosure purposes.

An unexpected improvement of an impact separation can be achieved by the arrangement and design of a separation opening. In this way, in particular, the gas volume provided in the impact separation can be reduced, so that the total volume of the gas-liquid separator can be reduced. This can surprisingly improve the separation efficiency of the chromatographic system.

A preferred gas-liquid separator comprises a separation area with an inlet nozzle, a baffle unit and a gas-conducting unit.

Preferably, the deposition area is designed so that an impact separation is effected. Impact separation means that the liquid droplets in the aerosol are directed against a baffle unit, whereby the liquid

Liquid droplets can form a liquid film.

As a baffle unit here can serve any body against which the aerosol stream can be passed. For example, the aerosol stream may be directed against an upper region of the separation region, for example against an upper termination of the deposition region. In this case, a protrusion, for example a spike or the like, may be provided on which the aerosol flow is applied, so that the liquid droplets directed onto the impact unit are not thrown back or rebound from the impact unit, but form a film.

A preferred gas-liquid separator uses gravity in operation, which causes separation of gas and liquid. Accordingly, the term above refers to the orientation of the gas-liquid separator that is in operation so that a gas can flow upwards, while down is the opposite direction, through which a liquid leaves the gas-liquid separator.

In a preferred embodiment, it can be provided that the baffle unit is substantially planar and can be regarded as a baffle plate, wherein this baffle plate preferably forms a wall of the separation region and a

Side wall of the gas guide unit represents. The expression "essentially flat" means that the baffle unit or the baffle plate is not bent, but may have a surface structure, In a specific embodiment, the baffle plate is preferably designed without a surface structure, so that this surface is smooth.

In a preferred embodiment, the baffle preferably comprises a surface structure, this embodiment being preferred over one having a smooth surface. Here, the surface structure preferably

Elevations and depressions, wherein the elevations with respect to the subsidence preferably a height in the range of 0.2 to 10 mm, more preferably in the range of 0.8 to 8 mm and particularly preferably in the range of 1, 5 to 5 mm show ,

Furthermore, it can be provided that the ratio of the height of the elevations (with respect to the subsidence) to the volume of the gas-liquid separator

preferably in the range of 0.01 to 10 mm / ml, more preferably in the range of 0.03 to 5 mm / ml.

In a further embodiment, the surface structure of the baffle unit has grooves, wherein the elevations and depressions of the grooves are preferably aligned in the direction of the, which is formed by the inlet nozzle and the Abscheideöffnung, or parallel to this direction.

By a structured surface of the baffle unit, preferably as

Grooved structure is formed, the volume of the gas-liquid separator can be kept particularly small and so the separation efficiency can be improved. In this case, the substances to be separated may have a relatively small difference in transit time, without thereby nullifying their separation in the gas-liquid separator. Furthermore, the degree of separation of the liquid can the aerosol with respect to the volume of the gas-liquid separator can be improved.

In a further development of the present invention, the baffle preferably has a surface area with a surface energy of at least 10 mN / m, more preferably at least 15 mN / m, and especially preferably at least 20 mN / m. Preferably, it can be provided that the baffle preferably has a surface area with a surface energy in the range of 15 to 120 mN / m, more preferably in the range of 20 to 80 mN / m, and especially preferably in the range of 22 to 60 mN / m preferably at least 80%, more preferably at least 90%, of the surface area of the baffle has a surface energy in the range of 20 to 80 mN / m, more preferably in the range of 22 to 60 mN / m. This surface energy can be achieved by an appropriate choice of material from which the baffle unit is made.

Furthermore, the baffle unit may have a surface area with a coating to adjust the aforementioned surface energy, wherein preferably at least 80%, more preferably at least 90% of the

Surface of the baffle unit has a coating.

The surface energy is determined by the method of Ownes-Wendt-Rabel & Kaelble. For this purpose, test series are carried out with the standard Busscher series, in which the test liquids used were water [SFT 72.1 mN / m], formamide [SFT 56.9 mN / m, diiodomethane [SFT 50.0 mN / m] and alpha Bromonaphthalene [SFT 44.4 mN / m] can be used. The measurement is carried out at 20 ° C. The

Surface energy can be determined with a contact angle measuring system G40 from Krüss, Hamburg, the implementation being described in the user manual of the contact angle measuring system G40, 1993. With regard to the

Calculation methods are to A. W. Neumann, About the measuring methodology for

Determination of Surface Energy Sizes, Part I, Journal of Phys. Chem., Vol. 41, pp. 339-352 (1964), and AW Neumann, About the Measurement Methodology for Determining Interfacial Energy Sizes, Part II, Journal of Phys. Chem., 43, p. 71-83 (1964). In addition to a baffle unit, an inlet nozzle is preferably provided in the separation region of the gas-liquid separator. Through the inlet nozzle, the aerosol is directed into the gas-liquid separator, in particular into the separation region of the gas-liquid separator.

In this case, inlet nozzle is preferably designed such that a through the

Inlet nozzle guided gas-liquid stream is acted upon against the baffle unit, as has already been set out above with regard to the baffle unit.

The shape and type of the inlet nozzle are not critical, so that it can be selected by the skilled person within the scope of his abilities. For example, the inlet nozzle may be designed such that the aerosol is directed onto the baffle unit in the form of a very narrow jet. Further, the inlet nozzle may also be designed to direct a cone-shaped puffing mist onto the baffle unit.

In this case, the nozzle can terminate with the wall of the deposition region or protrude over a projection into the separation region. The embodiment with a projection is advantageous if the baffle unit is provided in the upper end of the separation region.

Particularly preferably, the inlet nozzle is designed in the form of a simple bore or a simple opening. In a development, it can be provided that the inlet nozzle provided in the separation region has an entry surface that is substantially circular.

Furthermore, it can be provided that the space provided in the separation area

Inlet nozzle an entrance surface in the range of 0.05 mm ² to 20 mm ² , preferably in the range of 0.5 mm ² to 15 mm ² , more preferably in the range of 0.5 mm ² to 10 mm ^2, and especially preferably in the range of 0.8 mm ² to 5 mm ² . In a further embodiment it can be provided that the inlet nozzle provided in the separation region has an entry surface in the range from 2 mm ² to 40 mm ² , preferably in the range from 4 mm ² to 20 mm ² and especially preferably in the range from 5 mm ² to 15 mm ² has. This value refers to the size of a single inlet nozzle if multiple inlet nozzles are used.

If the inlet nozzle is formed in the form of a bore, this has

preferably a diameter in the range of 0.3 mm to 5 mm, preferably 0.5 mm to 4 mm, more preferably 0.8 mm to 3 mm, particularly preferably 1 mm to 2 mm and / or especially preferably 2 to 3 mm. This value refers to the size of a single inlet nozzle if multiple inlet nozzles are used.

Furthermore, it can be provided that the ratio of the entrance surface of the im

Abscheidegereich inlet volume to the volume of the gas-liquid separator in the range of 0.01 mm ² / ml to 1 mm ² / ml, preferably in the range of 0.04 mm ² / ml to 0.4 mm ² / ml, more preferably in the range of 0.08 mm ² / ml to 0.25 mm ² / ml, and more preferably in the range of 0.08 mm ² / ml to 0.17 mm ² / ml. This value refers to the sum of the areas of all used

Inlet nozzles, if several inlet nozzles are used.

Abscheidegereich inlet volume to the volume of the deposition range in the range of 1: 3 mm ² / ml to 1: 50 mm ² / ml, preferably in the range of 1: 5 mm ² / ml to 1: 20 mm ² / ml and especially preferably in the range from 1: 7 mm ² / ml to 1: 15 mm ² / ml. In a further embodiment it can be provided that the ratio of the entrance surface of the inlet nozzle provided in the separation region to the volume of the deposition region is in the range from 4: 1 mm ² / ml to 1: 50 mm ² / ml, preferably in the range of 1: 1 mm ² / ml to 1: 20 mm ² / ml and especially preferably in the range of 2: 3 mm ² / ml to 1: 5 mm ² / ml. This value refers to the sum of the areas of all inlet nozzles used, if several inlet nozzles are used.

In the separation area, one or more inlet nozzles can be provided. In the event that a plurality of inlet nozzles are provided, these are preferably aligned parallel. Preferably, the gas-aerosol mixture is passed through exactly one inlet nozzle into the deposition area, preferably to the baffle unit located in the separation area.

Furthermore, it can be provided that the inlet nozzle is configured such that a gas-liquid stream directed through the inlet can be acted upon against the baffle unit and the angle with which a gas-liquid stream conducted through the inlet nozzle can be acted upon by the baffle unit, preferably in the region of 50 to 130 °, more preferably in the range of 70 to 1 10 °. This angle can

be determined in particular by the direction of the inlet nozzle, with which the Inlet nozzle is directed to the baffle unit. This information refers to the angle at which the main jet of the aerosol is directed to the baffle. The shape of the aerosol jet is immaterial per se, as far as an impact separation can be effected. In this case, the liquid droplets of the aerosol should flow together by the impact on the baffle unit and preferably form a film. Therefore, the inlet nozzle should be chosen so that the liquid droplets of the aerosol do not become too small.

The gas-liquid separator preferably has a separation opening, which is arranged between separation region and separation region, so that there is a gas and liquid-open connection between these regions. By the

Separation opening is preferably effected an inertial separation. This means that the liquid flowing downwards at the baffle unit and / or the gas guiding unit in the form of a liquid film is separated from the gas by inertia. In this case, the gas preferably accelerates the liquid so that the liquid is transferred to the separation area at a higher speed than without this gas acceleration. This remains the

Liquid film preferably on a wall of the Abscheidebereichs, the

is preferably designed as part of the baffle unit and / or the gas-conducting unit, in the form of a film and passes directly into the separation area without the liquid film leaving this wall, which merges into the separation area. The

In contrast to the liquid phase, gas phase does not adhere to a wall, but is able to escape upwards and pass into the gas discharge area. In contrast, the liquid is discharged into the separation area and taken out of the gas-liquid separator via the liquid outlet provided in the separation area.

The shape of the separation opening is not critical as far as the previously stated function of the same can be fulfilled. Preferably, however, it can be provided that the deposition opening has an exit surface which is slit-shaped or has a plurality of openings arranged in parallel, which may be for example U-shaped, V-shaped or circular.

Preferably, the distance of the inlet nozzle from the baffle unit is greater than the smallest longitudinal extent of the separation opening. This results in the distance the inlet nozzle from the baffle unit from the path of the aerosol from leaving the inlet nozzle to impinging on the baffle unit. The smallest longitudinal extent of the separation opening relates to the width or length of the separation opening, wherein the extent of the plane up to the edge of the separation opening is related to the plane between separation area and separation area, which leads to a minimum area of the separation opening. In this level, in which the

Separation opening is located, the length of the longest extent of the

Determined separation opening, so that subsequently the shortest length of the

Abscheideöffnung can be measured, which is perpendicular to the longest dimension of the separation opening. This smallest length extension can also be considered herein as the width of the separation opening.

If the separation opening is slit-shaped, this preferably has a gap width in the range from 0.1 mm to 1.5 mm, particularly preferably 0.3 mm to 1.0 mm and especially preferably 0.4 mm to 0.7 mm (smallest linear expansion). The length of the gap is given by the circumference in the case of a circular or elliptical separation opening, these values preferably being in the range of 5 mm to 120 mm, particularly preferably in the range of 10 mm to 60 mm.

In a non-circular or non-elliptical separation opening having a gap shape, which is preferably characterized by two ends, its length is preferably in the range of 3 to 80 mm, preferably in the range of 5 to 50 mm, particularly preferably in the range of 15 to 30 mm.

If the separation opening is realized by a plurality of parallel openings, which may be for example U-shaped, V-shaped or circular, the dimensions set out above apply correspondingly, the openings preferably having a width in the range of 0.1 mm to 1.5 mm, more preferably 0.3 mm to 1, 0 mm and particularly preferably 0.4 mm to 0.7 mm (smallest length). In a further embodiment, the openings may preferably have a width in the range of 0.1 mm to 3.0 mm, particularly preferably 0.3 mm to 2.0 mm and especially preferably 0.4 mm to 1.5 mm (smallest linear expansion).

The gap width is measured perpendicular to the length or circumference of the gap and is the smaller longitudinal extent of the gap opening, which can be considered as a transition plane from the separation area to the separation area. These Transition plane has the smallest two-dimensional extent in the region of the transition from the separation region into the separation region.

Preferably, the deposition opening has an exit area in the range of 10 to 120 mm ² , more preferably in the range of 15 to 60 mm ^2, and particularly preferably in the range of 15 to 40 mm ² . In a further embodiment, the

Abscheideöffnung have an exit area in the range of 10 to 180 mm ² , more preferably in the range of 15 to 120 mm ^2, and more preferably in the range of 30 to 100 mm ² . Furthermore, it can be provided that the ratio of the exit surface of the separation opening to the volume of the gas-liquid separator in the range of 0.05 mm ² / ml to 2 mm ² / ml, more preferably in the range of 0.1 mm ² / ml to 1 mm ² / ml and especially preferably in the range of 0.3 mm ² / ml to 0.8 mm ² / ml. In a further embodiment it can be provided that the ratio of the exit surface of the separation opening to the volume of the gas-liquid separator in the range of 0.05 mm ² / ml to 6 mm ² / ml, particularly preferably in the range of 0.3 mm ² / ml to 3 mm ² / ml and especially preferably in the range of 0.5 mm ² / ml to 2.0 mm ² / ml.

The spatial form of the separation area is not critical and can be the

Be adapted to needs. In this case, a gas-conducting unit is preferably formed in the separation area. The gas guide unit causes a change in the

Flow rate of a gas, so that in the region of the inlet nozzle is a lower gas velocity than in the region of the separation opening. Since the volume flow in the same aerosol composition can be regarded as constant, this means that the aerosol is first passed into a relatively large space, which is then narrowed, so that the

Flow rate increases.

Accordingly, the cross-sectional area of the deposition area may be, for example, circular, for example from the inlet nozzle in the direction of

Separating opening is preferably narrowed in a wedge shape.

In a preferred embodiment, the deposition area has no

circular cross-sectional area in the region of the inlet nozzle, wherein the

Separation region preferably comprises at least three side walls, which define together with an upper termination of a space, which over the Separation opening is connected to the separation area. This embodiment, in which the separation region does not comprise a circular cross-sectional area, but has a cross-sectional area with corners, in particular a triangular, quadrangular, pentagonal or hexagonal cross-sectional area, particularly preferably a rectangular one, is easier to produce to a required precision, whereby the volume of the gas-liquid Separator can be better adapted to the requirements. In particular, gas-liquid separators can be provided which are suitable for particularly small volume flows. In contrast to gas-liquid separators with a circular cross-sectional area, gas-liquid separators with a non-circular, preferably one

Cross-sectional area with corners, exactly one inlet nozzle, without that

Areas of insufficient wetting with gas-liquid mixture would occur.

It can preferably be provided that the gas-conducting unit has at least two substantially planar side walls which can be regarded as gas-conducting plates, these gas-conducting plates preferably forming walls of the separating area. These two substantially planar sidewalls may converge to form a wedge shape.

Furthermore, it can be provided that the gas guide unit has at least two side walls, wherein at least one of the side walls is bent so that a concave shape is provided such that the two side walls can converge, wherein in the upper region of the deposition area, which through the Given the proximity of the inlet nozzle, the distance between the

Side walls is larger than in the lower part of the separation region, which is given by the proximity of the separation opening, wherein the decrease in the distance from the upper region to the lower region decreases.

Preferably, it can be provided that the gas-conducting unit has a

Gas acceleration unit, which together with at least one

Side wall, preferably at least two side walls causes a change in the flow rate of a gas.

In a further embodiment it can be provided that the cross-sectional area of the gas-conducting unit at least extends from the inlet nozzle in the direction of the separation opening partially decreases, preferably in the area facing the separation opening, so that the planes which are perpendicular to the flow direction of the gas-liquid mixture, smaller, this decrease is preferably continuous, so preferably at least two of the side walls of the gas guide unit in longitudinal section form a wedge shape ,

In a further embodiment it can be provided that the separation region comprises an upper termination, wherein this upper termination comprises a bend or an angle, wherein the highest point of the bend or the angle is preferably arranged centrally, and is thus in line with the inlet nozzle which are parallel to the gas flow direction or the direction of flow of the liquid, ie parallel to the direction of gas inlet liquid outlet opening, the upper end preferably merges into two side walls, so that the transition between the side walls and the upper end is curved.

In a further development of the present invention, it can be provided that the separation area comprises at least four side walls which, together with an upper termination, define a space forming the gas conduction unit, wherein one of the side walls is formed as a baffle unit, this space via the separation opening connected to the separation area. In this embodiment, in which the separation region comprises at least four side walls defining a space together with an upper termination, it may be preferred that the distance between two opposite side walls is greater than half the distance of the inlet nozzle from the baffle unit. Preferably, in this embodiment, in the separation area, at least four side walls are provided, which together with an upper termination define a space, the ratio of the distance between two opposite ones

Side walls at a distance of the inlet nozzle of the baffle unit in the range of 0.8 to 8, more preferably in the range of 0.9 to 6, more preferably in the range of 1, 0 to 4 and particularly preferably in the range of 1, 2 to 2 , These values relate in particular to two opposite side walls which have the greatest distance.

Furthermore, it can be provided that the inlet nozzle in the upper region of the

Abscheidebereichs is provided, particularly preferably in the upper third of Abscheidebereich, wherein this direction results from the arrangement of the inlet and the liquid outlet, so that the inlet nozzle is disposed above the liquid outlet.

In addition to the above-described deposition region, a gas-liquid separator according to the invention has a separation region. In the separation area, as previously indicated, the phases are separated, the separation area a

Liquid outlet, via which the liquid phase of the gas-liquid separator can be removed. The gas phase is directed into the gas discharge area. Accordingly, the separation region is connected to the gas discharge region via an opening and is in flow contact therewith.

Preferably, it can be provided that the separation region with a liquid outlet comprises a bottom, which preferably comprises a curvature, a curve, an angle or another shape, which leads to a taper, wherein the

Liquid outlet is provided in the region of the lowest point of the soil.

Furthermore, it can be provided that the liquid outlet is provided in the lower region of the separation region, particularly preferably in the lower third of the separation region

Separation area, wherein this direction results from the arrangement of the inlet nozzle and the liquid outlet, so that the inlet nozzle is arranged above the liquid outlet.

In a further embodiment it can be provided that the inner surface of the separation area has a surface area with a surface energy in the range from 15 to 120 mN / m, more preferably in the range from 20 to 80 mN / m and especially preferably in the range from 22 to 60 mN / m, wherein preferably at least 80%, particularly preferably at least 90% of the surface of the separation region has a surface energy in the range of 20 to 80 mN / m, particularly preferably in the range of 22 to 60 mN / m. Preferably, the difference of the surface energy of the inner surface of the separation area to

Surface energy of the inner surface of the deposition area at least

10mN / m, preferably at least 30mN / m be, these values refer to the respective maxima or minima, so that the difference is maximum. Furthermore, it can be provided that the separation region has a cross-sectional area in the region of the inlet nozzle which is at least 80%, preferably at least 90%, of the maximum cross-sectional area of the separation region, the cross-sectional areas being related to the planes perpendicular to the impaction unit and perpendicular to the main impact point of the separation nozzle Gas-liquid mixture opening,

The Gasausleitbereich serves to divert the gas phase from the gas-liquid separator, so that it comprises a gas outlet.

Preferably, the Gasausleitbereich is designed so that the

Gas velocity at the gas outlet is maximum, preferably the

Gas velocity in the gas flow direction from the separation area towards the gas outlet increases. In this way, a suction effect can be generated, which leads to a safe and low-maintenance operation of the gas-liquid separator. Furthermore, this can reduce the volume of the gas-liquid separator without its performance in other properties, such as the separation properties decreases.

In reverse of the separation area, therefore, the space decreases in direction

Separation area towards the gas outlet. Accordingly, the cross-sectional area preferably tapers from the separation area to the gas outlet.

In a further development of the gas-liquid separator, provision may be made for the gas discharge region to be such that the area of imaginary planes, which are perpendicular to the direction from the separation region to the gas outlet, decreases starting from the separation region towards the gas outlet, this decrease preferably is continuous, wherein preferably the gas-conducting unit forms a side wall of the Gasausleitbereichs and in longitudinal section this side of the gas-conducting unit with a further side wall of the Gasausleitbereichs a wedge shape.

Furthermore, it can be provided that the gas outlet in the upper region of

Gasausleitbereichs is provided, particularly preferably in the upper third of Gasausleitbereichs, this direction results from the arrangement of the inlet nozzle and the liquid outlet, so that the inlet nozzle above the

Liquid outlet is arranged. Furthermore, it can be provided that the inner surface of the gas discharge region has a surface area with a surface energy in the range from 10 to 40 mN / m, preferably at least 80%, particularly preferably at least 90%, of the surface of the gas discharge region having a surface energy in the range from 10 to 30 mN / m has.

In addition, it can be provided that the separation region is arranged above the separation region and the gas discharge region is arranged above the separation region, this direction resulting from the arrangement of the inlet nozzle and the liquid outlet, so that the inlet nozzle is arranged above the liquid outlet.

Furthermore, it can be provided that the deposition area above the

Separation region is arranged and the Gasausleitbereich is arranged above the separation region, this direction results from the arrangement of the inlet nozzle and the liquid outlet, so that the inlet nozzle is arranged above the liquid outlet.

Furthermore, it can be provided that the volume ratio of separation region to separation region is preferably in the range from 4: 1 to 1:10, preferably in the range from 2: 1 to 1: 6 and especially preferably in the range from 1: 1 to 1: 3 ,

In a further embodiment it can be provided that the

Volume ratio of separation region to Gasausleitbereich preferably in the range of 10: 1 to 1:10, preferably in the range of 5: 1 to 1: 5 and more preferably in the range of 2: 1 to 1: 2, is located.

Furthermore, it can be provided that the volume ratio of separation region to gas discharge region is preferably in the range from 10: 1 to 1: 4, preferably in the range from 6: 1 to 1: 2 and especially preferably in the range from 3: 1 to 1: 3 ,

Furthermore, it can be provided that the height of the deposition region is preferably in the range of 1 cm to 100 cm, particularly preferably in the range of 5 cm to 20 cm. Furthermore, it can be provided that the height of the separation region is preferably in the range of 0.5 cm to 20 cm, particularly preferably in the range of 2 cm to 5 cm.

Furthermore, it can be provided that the height of the gas discharge region is preferably in the range from 0.5 cm to 20 cm, particularly preferably in the range from 2 cm to 5 cm.

It can further be provided that the ratio of the height of the separation region to the height of the separation region is preferably in the range from 1: 2 to 10: 1, more preferably in the range from 1: 1 to 7: 1 and especially preferably in the range from 3: 1 to 6: 1 lies.

It can further be provided that the ratio of the height of the separation region to the height of the gas discharge region is in the range from 2: 1 to 1:10, particularly preferably in the range from 1: 1 to 1: 7 and particularly preferably in the range from 1: 3 to 1: 6 lies ..

The above-mentioned preferred properties of the gas-liquid separator require a definition of the various areas, each in a

Flow contact with each other, since the mixture of gas and liquid phase are transferred via the separation region in the separation region, in which the liquid is separated from the gas phase and the gas are transferred to the Gasausleitbereich. In this case, the separation opening forms the separation between separation area and separation area, wherein the plane in which the separation opening ends marks the transition to the separation area.

The transition between separation area and Gasausleitbereich is also marked by an opening, which is relatively large compared to the separation opening. This opening is defined by the plane which is arranged at the level of the separation opening and perpendicular to the direction of the gas flow direction of the gas-liquid mixture in the separation region or parallel to the flow direction of the gas phase, as soon as the separation of the

Separation region coming into the separation area merges or parallel to the liquid level during operation. The plane defined by the extent of the opening is selected to be the minimum area between

Separation area and Gasausleitbereich represents, this level the Abscheidideöffnung touched and is substantially parallel to the bottom of the separation area or during operation parallel to the liquid level.

Abscheidegereich inlet inlet and baffle inlet in the range of 5: 1 mm ² / mm to 1: 10 mm ² / mm, preferably 2: 1 mm ² / mm to 1: 5 mm ² / mm.

A preferred gas-liquid separator may be made of any known material as long as the requirements imposed by the solvents and the physical conditions are satisfied.

Preferably, a transparent material can be used, through which the separation process is visible, so that when a deposit or the like a rapid error analysis is possible.

Preferably, the gas-liquid separator may be made of metals, which are preferably acid and base resistant, from mineral glasses and / or plastics, for example fluoropolymers, polyetheretherketone (PEEK) or similar materials, which are preferably solvent resistant.

The gas-liquid separator preferably has a volume in the range from 20 ml to 100 ml, particularly preferably in the range from 20 ml to 70 ml, especially preferably in the range from 20 ml to 50 ml. In the case of a substantially parallelepiped shape which, for example, in the upper and / or lower region of the gas-liquid separator, which is provided through the inlet nozzle or through the liquid outlet, can be designed as an arc or a dome, the height of the gas-liquid Separator preferably in the range of 8cm to 150cm, more preferably in the range of 10cm to 12cm, the height through the

Length expansion in the gas flow direction, as from the inlet nozzle in the direction

Liquid outlet is given. The width and depth of the gas-liquid separator are each preferably in the range of 15mm to 60mm, more preferably in the range of 15mm to 25mm.

Preferably, it can be provided that the gas-liquid separator is not cylindrical, preferably not circular cylindrical, preferably one in the Substantially cuboidal basic structure having an upper and a lower arcuate cover.

The construction and production of a gas-liquid separator can be done in any way. According to a preferred embodiment, the gas-liquid separator can be configured separable, so that individual components can be assembled and disassembled. As a result, the gas-liquid separator can be easily cleaned when dirty. For example, an im

Substantially cuboid basic body can be produced with a suitable recess, on which a cover, which serves over a side wall

Gland be applied. The serving as a cover side wall can take over the function of the baffle and / or take over as part of the gas guide unit, as described above. A further part of the gas-conducting unit, which furthermore preferably constitutes a side wall of the gas discharge region, can be fastened in the substantially parallelepiped basic body with a suitable cut-out by means of a positive connection, by welding, preferably laser welding, gluing or the like, so that the above-described Areas, in particular at least one separation area, at least one separation area and at least one Gasausleitbereich arise.

The gas-liquid separator can generally be operated at atmospheric pressure. However, an accumulation of larger quantities

To avoid liquid, e.g. of methanol, the gas-liquid separator at a moderate internal counter-pressure of, for example, in the range of 0.1 bar to 4 bar operated by a back pressure regulator. Accordingly, can

be provided that the chromatography system after the gas outlet is provided a back pressure regulator, which is preferably in the range of 1 bar to 4 bar overpressure (absolute pressure 2 bar to 5 bar), preferably 2 bar to 3 bar overpressure adjustable. The collected over the separation area and by the

However, fluid component provided liquid component allows automated fractionation, which can be operated under atmospheric pressure. With the help of the gas-liquid separator and comparable to conventional HPLC analysis, a fully automatic fraction collection can also be realized for the SFC analysis. Since inner walls and components of the gas / liquid separator are substantially permanently moistened in the separation and separation areas, not only can a self-cleaning effect develop, but also a relatively low degree of cross-contamination of samples can be achieved. As a further advantage, the separator induces a relatively low peak broadening in the resulting chromatograms.

The gas phase of the aerosol can be collected and treated depending on the nature of the gas or, for example, when using C0 ₂ are released into the environment.

Furthermore, it can be provided that the chromatography system has a gas-liquid separator and a control valve is provided after the gas outlet of the gas-liquid separator.

The liquid phase of the aerosol is collected in a fraction collector. The collected fractions are particularly preferred automatically as

Main fractions collected while excess solvent one

Treatment or disposal may be subjected.

The connecting line between the liquid outlet of the gas-liquid separator and the fraction collector may preferably be designed so that residues of the gas phase, preferably C0 ₂ -Reste can escape via this compound. In particular, it can be provided that the chromatography plant has a gas-liquid separator and after the gas-liquid separator, a device for further separation of gas is provided, preferably a gas-permeable hose. For this purpose, a semipermeable plastic material may be used, for example Teflon, more preferably AF 2400 (commercially available from DuPont).

The number and size of collection vessels within the fraction collector are not subject to any particular limitation. Preferably, the

Fraction collector at least 5 collection vessels, preferably at least 10

Collection vessels, especially preferred at least 25 collection vessels. Furthermore, it can be provided that the fraction collector comprises collecting vessels, the one Have collection volume in the range of 5 to 500 ml, preferably 10 to 100 ml.

The fraction collector preferably comprises collecting vessels which have substantially the same collecting volume. A substantially the same

Collecting volume means, for example, that the collection volume of the individual collection vessels differs by at most 10 ml, preferably by at most 5 ml.

Furthermore, it can be provided that the collecting vessels of the fraction collector are designed as impact precipitators, wherein preferably the introduction of the fraction to be collected takes place at an angle in the range of 70 to 120 °, relative to the vertical, against a wall of the collecting vessel.

It can preferably be provided that the chromatography system comprises a UV detector. Furthermore, it can be provided that the chromatography system comprises a mass spectrometer as a detector. In a particularly preferred embodiment, the system comprises a UV detector and a mass spectrometer.

The SFC chromatographic system is preferably operable at a flow rate in the range of 10 ml / min to 450 ml / min, more preferably in the range of 50 ml / min to 300 ml / min, and especially preferably 100 ml / min to 250 ml / min. Furthermore, it can be provided that the SFC chromatography system

preferably at a flow rate of at least 10 ml / min, more preferably of at least 50 ml / min and especially preferably of at least 100 ml / min is operable.

Surprisingly, the process according to the invention can also be carried out with conventional HPLC plants after manageable conversions.

In another aspect, a conversion kit is also provided by which a high performance liquid chromatography (HPLC) system can be converted to an SFC system. Such a kit preferably comprises at least one gas-liquid separator as described above. Preferably, the kit contains another component, as described below, to convert a HPLC plant into an SFC system, such as heat exchangers or back pressure regulators.

In many cases, the control unit of a HPLC system is not freely accessible, but on the input of conventional parameters, such as flow rate, volume of

Collecting vessels of the fraction collector, time span, the one to be collected

Liquid is needed to measure the distance between the detector signal and the

Limit fraction collection valve, etc. limited. A variable flow rate, as this results from a preferred separation of the second solvent at the fraction collector, can often not be programmed into the control unit of a HPLC system. To avoid this difficulty, a programmable logic controller (PLC unit) may preferably be provided.

In a preferred embodiment, a second control unit, for example a programmable logic controller (PLC unit), is furthermore provided.

provided in operative connection with the control unit of the HPLC system, the pump, in particular the pump for the first solvent and / or the

Fraction collector of this plant stands. Furthermore, the second control unit, for example the SPS unit, can be connected to the detection unit, for example a mass spectrometer and / or a UV detector, in order, for example, to send a control signal to the control unit of the HPLC system.

For programming a variable flow rate in a designated controller, the flow rate of the first solvent can preferably be determined, which can be done for example by measuring the first solvent flow before or after the solvent pump with a flow sensor. Furthermore, pumps can be used which allow a readout of the signals provided by the chromatography software. The corresponding signals are passed to the second control unit, for example, the PLC unit and processed by the latter.

In a further step, the system dead volume between the pump and the fraction collecting valve is preferably determined. This dead volume creates a

System delay time of incoming solvent composition at Fraction collecting valve in gradient mode. The dead volume consists of the inner volume of all Kappilaren / piping between the pump and

Fraction collection valve, the internal volume of the chromatography column (stationary phase porosity must be taken into account). To determine the dead volume between the pump and the fraction collection valve, a UV-absorbing solvent can be used, for example, as the second solvent

Gradients that can be determined on the UV detector, and can be collected accordingly in the fraction collector, the dead volume or the dead time can be determined. Furthermore, the dead volume can be roughly estimated and based on this estimate a first test run can be carried out, with all fractions being collected (threshold value = 0). Based on the filling level of the individual collecting vessels, the dead volume or the dead time can then be optimized. If the fill level was too low, the dead volume or the dead time was estimated to be too low; if the fill level was too high (in particular if it overflowed), the dead volume became

or the dead time estimated too large.

To carry out this modification of the present method, the second control unit, for example, the PLC unit of the control unit of the HPLC system provides a control signal, this depending on the concrete

Embodiment can be made. For example, the second control unit, for example the PLC unit, can be connected to the control unit of the HPLC system via an input for a further detector or via a corresponding interface. Furthermore, in many cases between the signal of a detector, preferably a UV detector and the control unit of the HPLC system, the second control unit, such as the PLC unit are interposed, so that the signals of the detector first in the second control unit, such as the PLC unit be guided and from the second control unit, for example, the PLC unit then in the control unit of the HPLC system.

Further, in order to practice this variation of the present method, the volume value of a collecting vessel of the fraction collector should be set correspondingly high to prevent premature switching by the control unit of the HPLC plant. For example, for an actual volume of 20 ml, the volume value of a collector of the fraction collector of at least 300 to 400 ml, depending on the proportion of the first solvent.

In a modification of a conventional HPLC system, which does not allow comprehensive access to the control, in particular the control of the fraction collector via a second control unit, which is designed for example as a PLC unit, at least partially, wherein the second control unit, such as the PLC - Unit is in operative connection with the fraction collector and preferably the second control unit, for example, the PLC unit data on the

Pumping or the flow rate and the gradient are provided.

Further, the second control unit, for example, the PLC unit

Detector data, preferably provided signals of a UV detector.

The control of the fraction collection valve is preferably at least partially configured via the control unit of the HPLC system. Here is the

Signal recognition of a signal detected by a detector as a fraction to be collected preferably realized by the control unit of the HPLC system, while a change of the collecting vessel preferably via the second control unit, which is designed for example as a PLC unit, is effected.

The signal recognition can take place, for example, via a threshold value (signal height) and the slope of the signal curve or other factors of the signal detected by the detector and transmitted to the control unit of the HPLC system, whereby it correspondingly switches the fraction collection valve and fills the collecting vessel. In general, the collection vessel is filled until the signal no longer meets the criteria, for example, a detected peak falls below the intended threshold. If the collection time for this peak is longer than the specified volume of the collection vessel, the collection process is interrupted for a short time in order to continue the collection process at the next empty collection vessel.

The problem here is, as already stated, that a conventional HPLC software can not consider a flow gradient, according to the preferred

Embodiments of the fraction collection valve are present, since preferably the second solvent is mainly separated and to be collected

Compositions predominate containing the first solvent. The calculation of the maximum filling level by the control unit of the HPLC system is preferably avoided by the fact that in the software for controlling the HPLC system a fraction collection volume is deposited, which

for example, 10-20 times larger than that of the real fraction glasses. The

However, signal detection (peak detection) is preferably unaffected in this case.

Fraction end still takes place via the software of the control unit of the HPLC system. Since, in preferred embodiments, a steadily increasing flow gradient is applied to the fraction collection valve, the maximum fraction collection time decreases correspondingly. The maximum fraction collection time is therefore preferably provided by a second control unit, for example a PLC unit.

For this purpose, the flow rate is determined, which can be done according to the above method. The gradient of the first solvent is preferably in a

Memory array of the second control unit, such as the PLC unit periodically stored. Depending on the resolution, you can use any number of memory lines. Good results can be achieved, for example, at a resolution of one memory line per second.

This relationship can be represented, for example, by the following formula (II):

Formula (II) where:

F _L i (t) is the volume flow of the first solvent at time t (measured at or calculated for the fraction collection valve)

AA _S is the time period over which the volume of a single volume component is computed, resulting from the resolution of the storage array. The resolution of the memory array results from the number of Memory cells and the dead time of the system (running time between pump outlet and fraction collection valve).

V _max is the maximum volume of the collection vessel t _s is the start time at which the detector signal indicates a fraction to be collected and to which t _n is the end time to which a fraction to be collected enters

Collection vessel can be initiated

Accordingly, the total volume is continuously calculated from the sum of the individual volume components and at the time of this

For explanation, the following numerical example is given, without this being intended to limit it:

With a memory line resolution of 1 s and a system delay time of 60 s, a memory array of 60 lines is required. In this memory array thus always the current Gradientverlauf the last 60s is stored and updated every second. That the memory value of the 60s is "old" is overwritten with the current flow value, however, the overwriting only takes place after this value has been read, for example, for the calculation of the collection volume.

From the moment when the fraction collection valve switches, the PLC (via the control voltage of the valve) receives the signal for integrating the

accumulated volume in the fraction glass. The gradient profile is taken into account. That from the "oldest" memory line, which is the share of the first

Solvent corresponds to the fraction collection valve, is accurate to the second summed up and compared with the deposited fraction volume. If the totalized fraction volume is equal to the deposited fraction volume (stored in the PLC), a signal is sent by the PLC to the chromatography software for changing the fraction collection vessel. The type and length of this signal depends on the HPLC system to be rebuilt. For example, a corresponding signal can be sent via the input of the UV detector to the HPLC control unit, which is for example 500 ms long.

The chromatographic software has previously programmed a corresponding trigger ("trigger") so that the UV detector and "PLC detector" are logically linked.

This results, for example, in the following relationships:

T _sys = system delay time = 60s n _sz = number of memory lines = 60

Memory clock: 1Hz (every second a new memory line is written)

T _SYS 60 s

A _s = resolution memory array = = = ls

_sz 60

F (t) _G = flow at time t in the gradient curve in ml / min

F (t) c

V _s = memory line volume = ^ ^■ A _s V _max = maximum fraction volume can be summarized following formula

If the HPLC system to be rebuilt has a significant delay between switching signal and actual switching on the fraction collection valve, this switching delay must be taken into account when calculating the volume. This is often the case, if the rebuilt HPLC system For example, equipped with a mass spectrometer. The evaluation of the mass spectrometry signal generally requires a relatively large period, which is often in the range of 1 s to 10 s, for example about 4 s to 6 s. In many cases, an additional volume is therefore provided in these systems, as previously and subsequently explained in more detail.

Preferably, it may be provided that the degree of filling of the collecting vessel takes place via a summation of the volume, which over a period in the

Collection vessel is introduced, wherein the individual volume components of the summation on the products of the flow rate, the period over which the volume component is collected, and the proportion of the first solvent in the solvent composition at the start time of the respective

Volume component is determined, taking the time to determine the

Mass spectrometry signal is required by an extra volume in the

Summation is taken into account, wherein the volume of the collecting vessel is stored in the control unit.

This relationship can be represented, for example, by the following formula (III):

Formula (III) where

F _L i _(t) is the volume flow rate of the first solvent at time t (measured at the fraction collection valve

AA _S is the time period over which the volume of a single volume component is computed, resulting from the resolution of the storage array. The resolution of the memory array results from the number of memory cells and the dead time of the system (running time between

Pump outlet and fraction collection valve). V _max is the maximum volume of the collection vessel t _s is the start time at which the detector signal indicates a fraction to be collected and to which t _n is the end time to which a fraction to be collected enters

Collection vessel can be initiated

T _v is the delay period, ie the time interval between the switching signal and the actual circuit at the fraction collection valve

Accordingly, an additional volume for calculation over formula (II) is provided, which is calculated continuously.

This results, for example, in the following relationships:

Assumption: 6s signal delay time mass spectrometer

Today's mass spectrometers have a signal processing time of up to 6s compared to the UV detector signal.

If the HPLC system to be rebuilt has a signal delay time of 6s, the PLC signal must be output 6s earlier, otherwise the fraction glass would overflow the volume of the 6s. At the faction start is at the

Volume calculation adds the volume of the following 6s from the beginning to it. The gradient curve is permanently monitored for the period of the 6s.

With the symbols set out above, this can be summarized to the following formula:

In this case, F _(t ) currently corresponds to the respective flow of the first solvent, where F _(t ) currently corresponds to the end F (t) G end. Hereinafter, preferred embodiments of the present invention will be described by way of example with reference to four figures, without this being intended to limit the invention. Show it:

Figure 1 is a schematic representation of a chromatographic system for

Implementation of the present method.

Figure 2 is a schematic longitudinal sectional view of a preferred

usable gas-liquid separator,

Figure 3 is a schematic cross-sectional view of a preferred

usable gas-liquid separator,

FIG. 4 is a schematic plan view of a preferably usable gas-liquid separator;

Figure 1 shows a schematic representation of a chromatography system 100 with a gas-liquid separator 130 according to the invention, which for a

supercritical fluid chromatography is suitable.

Such a system is exemplified using supercritical CO ₂ , illustrating methanol as an exemplary solvent. Of course, systems in which other solvents, preferably organic solvents are used, or other supercritical fluids are used, similarly constructed.

As shown in Fig. 1, the respective fluids are stored in storage tanks

especially in a supercritical state

used gas in a storage tank 102 and the solvent one

Storage tank 104 is provided, which can be funded via a respective pump 106, 108 from the storage tanks 102, 104 to the other components of the system. In the system 100 described herein, a preparation stage 1 10, 1 12 is preferably provided in each fluid supply line, via which the liquids can be tempered. Furthermore, a leveling of the pressure fluctuations indicated by the pumps can be provided. Accordingly, this preparation stage may be formed, for example, as a heat exchanger or as a pump. In the solvent line, an addition unit may preferably be 14 be provided, for example, an injector over which a aufzutrennende

Mixture be introduced into the system 100 before the C0 ₂ and the

Solvent passed into a mixer 1 1 6 and from this one

Chromatography column 1 18 are supplied.

In the present system 100, the chromatographic column 1 18 are two

Downstream analysis units, for which purpose a Probeausleiteinheit 120 is connected to a mass spectrometer 122 and after the Probeausleiteinheit a UV detector 124 is provided. According to the analysis unit, a device for providing an additional volume 125 is set forth here, which serves in particular to increase the transit time of the liquid in order to be able to evaluate results of, for example, the mass spectrometer 122. The back pressure regulator 126 provided in the conduit downstream of the additional volume provision means 125 maintains the respective pressure necessary for the fluid to remain in a supercritical state. After this

Back pressure regulator 126 is a heat exchanger 128 is provided which prevents freezing of the aerosol during the expansion process. Subsequently, the aerosol is introduced into a gas-liquid separator 130 according to the invention, wherein the gas of the system is discharged via outlet 132.

The liquid is introduced into a fraction collector 134 and fractionated therein. The solvent contained in the fractionated samples can be removed from the samples.

Figure 2 describes a preferably usable gas-liquid separator 10 in a longitudinal sectional view.

The gas-liquid separator 10 comprises a separation region 12 with an inlet nozzle 14, a baffle unit 1 6 and a gas-conducting unit 18. The gas-conducting unit 18 is formed by the baffle plate 16 constructed here as a baffle plate

Gas acceleration plate 20 and not shown in longitudinal section further two side walls formed. Shown is in particular the present wedge-shaped form of the separation region 12, through which a gas is accelerated from the region of the inlet nozzle 14 toward the separation opening 22. The baffle plate 1 6 formed here as a baffle plate may have a structured or smooth surface. The gas acceleration plate 20 may be flat from the direction of the inlet nozzle 14 in the direction of the separation opening 22 or slightly curved concavely, so that the presently apparent decrease in the distance between the baffle plate 1 6 and the gas acceleration plate 20 is reduced. At the top, the separation region 12 is delimited by an upper termination 24.

The gas-liquid separator 10 comprises a separation region 26 with a

Liquid outlet 28, wherein the separation region 26 is connected to the separation region 12 via the separation opening 22, so that the separation region 12 is in flow contact with the separation region 26.

In the present case, the baffle plate 1 6 formed as a baffle plate forms a side wall of the separation region 26. The bottom of the gas-liquid separator 10 is formed by the lower end of the separation region 26. This floor can be designed so that the liquid outlet 28 is provided at the lowest point of the floor.

A side wall 32 of the Gasausleitbereichs 30 and the two side walls not shown in longitudinal section together with an opening 34 which is provided between Gasausleitbereich 30 and the separation region 26 and the Abscheideöffnung 22, the other boundaries of the separation area.

In the separation region 26, the gas phase is separated from the liquid phase, wherein preferably the gas is accelerated by the gas guide unit 18 in the direction of the separation opening 22, so that the liquid is transferred in the direction of the bottom of the separation region 26.

The gas phase is conducted into the gas discharge region 30 via the opening 34, which is provided between gas discharge region 30 and the separation region 26. The

Gas discharge region 30 is in the present case designed such that the gas is accelerated in the direction of gas outlet 35, which is provided in gas discharge region 30.

In the present case forms the rear wall of the previously described gas acceleration plate 20 together with the protruding into the separation area side wall 32 a

corresponding wedge shape, wherein an edge of the gas acceleration plate 20 is connected to the side wall 32. Figure 3 shows a schematic cross-sectional view of a preferably usable gas-liquid separator 10, wherein like reference numerals describe like parts.

In particular, the side walls 36, 38 of the gas-liquid separator 10 (not shown) are shown. Furthermore, the supply line 40 of the aerosol and the discharge line for the gas 42 are shown.

It can also be seen that in this embodiment, the baffle plate

formed baffle unit 1 6 has a groove-shaped surface structure.

Figure 4 shows a schematic plan view of a preferably usable gas-liquid separator 10, wherein like reference numerals describe like parts. In particular, the preferred embodiment of the lower end 44 of the separating region 26 and of the upper end 24 of the separating region 12, which in the present case are configured in an arc shape, is apparent.

The features of the invention disclosed in the foregoing description, as well as the claims, figures and embodiments may be essential both individually and in any combination for the realization of the invention in its various embodiments.

Claims

claims

1 . Method for operating a fractionation collector in a

Chromatography, wherein a solvent composition in a

Chromatography column is pumped, which contains at least a proportion of a first solvent which is liquid under normal conditions, and contains a proportion of a second solvent which is gaseous under normal conditions, and the flowing from the chromatographic column

Composition is at least partially introduced into a detector, characterized in that a composition containing the first solvent is introduced after leaving the chromatography column in response to the detector signal in a collecting vessel of the fraction collector and the amount of liquid which is introduced into a collecting vessel of the fraction collector, is selected depending on the proportion of the first solvent.

2. The method according to claim 1, characterized in that in a

Chromatography column pumped solvent composition is changed during the course of chromatography.

3. The method according to claim 1 or 2, characterized in that the

The proportion of the first solvent in the solvent composition is determined by the time of the start of the fraction collection, wherein preferably a control unit over the course of time provided solvent composition is input and based on the start time of the collection, the proportion of the first solvent is determined and based on this proportion and the Flow rate is set the collection time for a collection vessel, wherein in the control unit the

Volume of the collecting vessel is deposited.

4. The method according to any one of the preceding claims, characterized in that the degree of filling of the collecting vessel via a Summation of the volume that occurs over a period in the

Collection vessel is introduced, wherein the individual volume components of the summation over the products of the flow rate, the period over which the volume component is collected, and the proportion of the first solvent in the solvent composition is determined at the start time of the respective volume component, wherein in the

Control unit, the volume of the collecting vessel is deposited.

Method according to at least one of the preceding claims, characterized in that the second solvent which is used in the

Composition is contained after the chromatographic column, at least partially separated before being introduced into the fraction collector.

Method according to at least one of the preceding claims, characterized in that a gas-liquid separator is used and the pressure in the gas-liquid separator in the range of 0.1 bar to 4 bar,

preferably 0.5 bar to 3.5 bar and particularly preferably 1 bar to 3 bar (overpressure).

Method according to at least one of the preceding claims, characterized in that the fractionation is operated at a lower pressure than the gas-liquid separator, wherein preferably the pressure difference in the range of 0.1 bar to 4 bar, preferably 0.5 to 3 bar , 5 bar and more preferably 1 bar to 3 bar.

Method according to at least one of the preceding claims, characterized in that the fraction collector is controlled by a controller and the controller is in operative connection with the detector, wherein after completion of the detection of a substance by the detector

Control pulse is passed to the fraction collector, which causes a change of the collecting vessel.

9. The method according to any one of the preceding claims, characterized in that in determining the timing of the collection start for the initiation of the fractions to be collected, the liquid volume of the lines and devices between the detector is taken into account until the separation point, wherein the fractions to be collected by the fractions to be discarded.

10. Chromatography system for carrying out a method according to

at least one of the preceding claims, characterized in that the Chromatorgraphie plant comprises a controller that in

Actively connected to a detector and a fraction collector and the controller is programmable so that the amount of liquid that can be introduced into a vessel of the fraction collector, depending on the proportion of the first solvent is fixable.

1 1. Chromatography system according to claim 10, characterized in that the fraction collector comprises collecting vessels which have substantially the same collecting volume.

12. Chromatography system according to claim 10 or 1 1, characterized

characterized in that the collection vessels of the fraction collector as

Prallabscheider are configured, preferably the introduction of the fraction to be collected at an angle in the range of 70 to 120 °, based on the vertical, takes place against a wall of the collecting vessel.

13. Chromatography unit according to at least one of the preceding

Claims 10 to 12, characterized in that the chromatography plant comprises a UV detector.

14. Chromatography plant according to at least one of the preceding claims 10 to 13, characterized in that the chromatography plant has a gas-liquid separator and after the gas outlet of the gas-liquid separator, a control valve is provided.

15. conversion kit for converting a high-performance liquid chromatography system to a chromatography system according to any one of the preceding claims 10 to 14, characterized in that the kit comprises at least one gas-liquid separator and at least one second control unit.