Kit, pre-purification and injection device and pre-purification and injection method for pre-purifying sample of radiolabeled compound in order to purify sample by high pressure chromatography

Abstract

The invention relates to a kit for pre-purifying a liquid sample comprising a radiolabelled compound intended to be purified by high pressure chromatography, comprising: a housing (2), filtering means (3), a chamber (4) comprising a solid stationary phase, and fluid communication means (5), the housing (2) and the fluid communication device (5) can mechanically withstand a fluid pressure greater than or equal to 50 bar. The invention also relates to a pre-purification and injection device comprising said kit, and to a pre-purification and injection method using said kit.

Description

[0001] The present invention relates to the field of synthesis and purification of radiolabeled compounds, such as injectable radiolabeled tracers for positron emission tomography (PET) analysis. In particular, the present invention relates to a kit for prepurifying a raw liquid sample obtained by synthesis of a radiolabeled compound in order to analyze / separate / purify it by high pressure chromatography. The present invention also relates to an apparatus and a method for prepurifying and injecting a liquid sample of a radiolabeled compound into a high pressure chromatography system.

[0002] The synthesis (or radiosynthesis) of radiolabeled compounds is usually based on "wet" chemistry, in which radiosynthetic units react with non-radioactive precursors in solution in a synthesis module or radiosynthesizer. Since the radioactive compounds are intended to be injected into patients, this method necessarily requires steps involving purification of the synthesis product (the original sample containing the radiolabeled compound).

[0003] To purify the original sample from the radiosynthesis, a known and commonly used method is high performance or high pressure liquid chromatography (HPLC). The purification step must then be followed by a reconstitution step in a biocompatible solvent, since commonly used HPLC solvents (such as acetonitrile) are not suitable for injection into humans or animals.

[0004] Supercritical fluid chromatography (or SFC) is also a well-known method when purifying raw samples of synthetic radiolabeled compounds, but its application is less extensive and is mainly used for compounds labeled with carbon-11 and compounds with low levels of radioactivity. This method uses a supercritical fluid as the mobile phase (usually carbon dioxide), usually in combination with a co-solvent or "modifier" (such as ethanol) that changes its polarity. Compared to HPLC, the separation step of SFC is faster (at least two to three times that of HPLC), which is a significant advantage for short-lived radioisotopes because it increases the final synthetic yield. In addition, SFC eliminates the need for reconstitution because the purified sample can be recovered in ethanol, which is suitable for injection into the body after simple dilution (for example with saline or aqueous buffer).

[0005] In both purification methods, the raw sample of the radiosynthesis is usually injected into the chromatographic column through an injection system consisting of an injection loop, which allows a maximum volume equal to the volume of the loop to be injected into the column (in the case of using the entire volume of the loop, it is also possible to guarantee the reproducibility of the injection volume), while the valve allows the fluidic communication of the different elements ("outlet" of the synthesizer, the loop, the pump, the "waste" container, etc.). The disadvantage of such a loop is that it limits the volume delivered to at most the volume of the loop, unless the filling and injection of the loop takes place several times (automatically), which is very time-consuming and should therefore be absolutely avoided in the case of radiosynthesis. Moreover, in this case there is always the risk of yield losses due to overflowing, etc.

[0006] In particular, HPLC methods can be used to manage high injection volumes (e.g. up to 10 mL), thus enabling a certain speed to be achieved, since the loop is only loaded once, and limiting yield losses (due to loss of activity in the radiosynthesizer), since one or more flushing operations can be performed to maximize the recovery of the original sample from the synthesis of the radiolabeled compound. On the other hand, the resulting dilution can reduce the chromatographic performance and affect the purity of the final product.

[0007] Furthermore, under standard conditions, the SFC method requires a low-volume injection loop (less than 1 mL) because larger injection volumes induce a dramatic decompression when the injection valve is reset to the loading position. Therefore, this separation method is not suitable for large-scale (i.e., volumes greater than 1 mL) syntheses.

[0008] However, the configuration of conventional radiosynthesizers does not allow automatic transfer without loss of radioactivity in volumes less than 1 mL. For original sample volumes greater than 1 mL, in order to be able to use SFC and benefit from its above-mentioned advantages, (pre-)concentration, for example by evaporation of the solvent, can be considered, but in order to limit losses, manual injection of small volumes of original sample into the loop is required, which is unthinkable for the amounts of radioactivity used in production. Moreover, in the case of short-lived radioisotopes, for example, an additional evaporation step will automatically affect the synthesis yield.

[0009] More specifically, there are also separation systems using the SFC (industrial preparation) method with larger injection loops (e.g. 5 mL), but these systems are bulkier and very expensive. Therefore, they have little application in the synthesis of radiolabeled compounds.

[0010] Furthermore, due to the risk of encountering chromatographic column clogging problems, it is usually necessary to first filter the raw sample obtained by radiosynthesis, which again constitutes an additional step that prolongs the time of the entire synthesis-purification process. In addition, this filtration step requires a strict selection of the filter to minimize losses on the filter membrane (e.g., through chemical interactions). An object of the present invention is to address the above-mentioned disadvantages of the prior art.

[0011] In particular, it is an object of the present invention to provide a device and a method for prepurifying and injecting a raw sample obtained from the synthesis of a radiolabeled compound into a high pressure chromatography system, which reduces the time required to obtain a purified radiolabeled compound compared to prior art systems, thereby obtaining a higher final yield.

[0012] It is also an object of the present invention to provide an apparatus and a method for prepurifying and injecting a raw sample obtained by synthesis of a radiolabeled compound into a high pressure chromatography system, regardless of the initial volume of the sample.

[0013] Another object of the present invention is to provide an apparatus and a method for prepurifying and automatically injecting a raw sample obtained by synthesis of a radiolabeled compound into a high pressure chromatography system without manual operation.

[0014] Another object of the present invention is to provide a device and a method for prepurification and injection of raw samples obtained by synthesis of radiolabeled compounds, which device and method allow one or more flushing of the radiosynthesizer (thus optimizing the yield of the radiosynthesis by minimizing losses in the radiosynthesizer) without increasing the volume injected onto the column.

[0015] Another object of the present invention is to provide a device and a method for pre-purifying and injecting a raw sample obtained by synthesis of a radiolabeled compound into a high pressure chromatography system, which device and method can control the injection volume regardless of the initial volume of the sample, thereby allowing better chromatographic reproducibility.

[0016] Another object of the invention is to provide a device and a method for prepurification and injection of raw samples synthesized from radiolabeled compounds, which allow a single (automatic) injection into the chromatographic system regardless of the initial volume of the sample, thus saving time. Another object of the invention is to provide a device and a method for prepurification and injection of raw samples synthesized from radiolabeled compounds, which also allow filtration, thus preventing any subsequent clogging of the chromatographic column.

[0017] To achieve these objects, the present invention provides a kit for prepurifying a liquid sample comprising a radiolabeled compound intended to be purified or analyzed by high pressure chromatography.

[0018] In particular, the kit of the present invention comprises:

[0019] -shell,

[0020] - filtration devices,

[0021] - a chamber, the volume of which is defined by the housing and the filtering device, said chamber comprising a solid stationary phase, and

[0022] - fluid communication means located upstream and downstream of the chamber and the filtering means,

[0023] The housing and the fluid communication arrangement are mechanically capable of withstanding a fluid pressure greater than or equal to 50 bar.

[0024] The present invention also provides a device for prepurifying and injecting a liquid sample including a radiolabeled compound into a high pressure chromatography system.

[0025] In particular, the device of the present invention comprises:

[0026] -Multi-way valve,

[0027] - means for supplying a liquid sample comprising a radiolabeled compound,

[0028] - elution phase pump,

[0029] - a kit according to the invention,

[0030] - discharge device,

[0031] The valve is in fluid communication with the supply, pump, reagent cartridge, drain, and high pressure chromatography system.

[0032] Finally, the present invention also provides a method for prepurifying and injecting a liquid sample comprising a radiolabeled compound into a high pressure chromatography system.

[0033] Specifically, the method of the present invention comprises in sequence:

[0034] (i) a loading step, comprising:

[0035] - supplying the reagent kit according to the invention with a liquid sample comprising a radiolabeled compound via a multi-way valve located in a loading position,

[0036] -retention of the radiolabeled compound by the kit,

[0037] - removing the components not retained by the reagent kit to the discharge device through the multi-way valve;

[0038] (ii) an injection step, comprising:

[0039] - the elution phase is pumped by a pump through the multi-way valve in the injection position towards the reagent cartridge,

[0040] - releasing the radiolabeled compound retained by the kit,

[0041] - The elution phase comprising the radiolabeled compound is introduced into the high pressure chromatography system via a multi-way valve.

[0042] The present invention is therefore based on a new and inventive approach. In particular, the inventors surprisingly found that by replacing the injection circuit in a conventional injection device in a high pressure chromatography system (HPLC type or SFC type) with a kit comprising the features of the present invention, it is possible, whatever the initial volume of the sample comprising the radiolabeled compound, to significantly increase the synthesis yield by (i) significantly reducing the separation time (up to 4 times) and (ii) allowing flushing of the radiosynthesizer (thus minimizing the loss of compound in the radiosynthesizer) without increasing the volume injected onto the column. For the sake of clarity, it is therefore understood that the conventional injection circuit in a high pressure chromatography system is not included in the device of the present invention, or in other words, the device of the present invention does not include an injection circuit.

[0043] In addition, the present invention can also avoid manual operation and realize automatic injection.

[0044] Other features, details and advantages of the present invention are contained in the description and drawings given below, but are not limited thereto.

[0045] In the present specification and claims, it is clearly understood that, unless explicitly stated otherwise, the terms "a", "an" or "the" used herein mean "at least one" and should not be limited to "single". The terms "comprise", "include", "contain" and "have" have an open meaning and do not exclude the presence of other components. In addition, when a numerical range is indicated, the lowest and highest values ​​are included. Finally, all integer values ​​and sub-domain values ​​in the numerical range are explicitly included, just as if they were explicitly written out.

[0046] According to the invention, the kit allows the pre-purification of a liquid sample comprising a radiolabeled compound intended to be purified or analyzed by high pressure chromatography.

[0047] According to the invention, the pre-purification of the liquid sample may also correspond to or comprise the concentration of the sample, in particular the concentration of the radiolabeled compound.

[0048] In particular, the sample according to the invention may comprise one type of radiolabelled compound or several types of radiolabelled compounds. Radiolabelled compounds according to the invention are, for example, organic molecules carrying radioisotopes (e.g. carbon-11, fluorine-18, iodine-131, yttrium-90 or actinium-225), for example (i) sugars (e.g. glucose) or amino acids.

[0049] According to the present invention, the liquid sample comprises a radiolabeled compound, which is intended to be purified or analyzed by high pressure chromatography. The kit of the present invention is in fact suitable for analytical chromatography (e.g., the presence or formation of one or more compounds can be detected) or preparative or semi-preparative chromatography (one or more purified compounds can be collected from the original sample obtained by synthesis).

[0050] According to the present invention, the liquid sample can be a sample in an aqueous phase or an organic phase or two phases (aqueous phase / organic phase) or an emulsion. Preferably, the liquid sample is a single-phase sample in an aqueous phase. The aqueous phase according to the present invention comprises at least 40% (by volume) of water, preferably at least 50% of water, or even at least 70% of water.

[0051] According to the invention, the kit comprises a filtration device. The filtration device according to the invention can (through the housing) retain / capture the solid stationary phase in the chamber by allowing the fluid to pass through and filtering the original sample of potential solid particles (thereby preventing any clogging of the chromatographic column during purification by high pressure chromatography).

[0052] Preferably, the filtering means is selected from a filter membrane and a glass frit.

[0053] According to the present invention, the kit comprises a chamber whose volume is defined by the housing and the filtering device. Preferably, the volume of the chamber is 50 mm 3 Up to 2000mm 3 Most preferably, the volume of the chamber is between 50 mm 3 Up to 1000mm 3 These volumes are particularly suitable for liquid samples obtained by radiosynthesis.

[0054] According to the invention, the chamber comprises a solid stationary phase. "Solid stationary phase" refers to the generally accepted definition in the field of separation / chromatography of chemical components in a mixture: "solid stationary phase" is a compound in solid form used to retain / adsorb the components of the mixture to be separated. This phase is immobile, in contrast to the mobile phase (liquid or gas) that migrates through the stationary phase. The adsorptive interactions that occur between the solid stationary phase and the components can be of various types: hydrogen bonding, hydrophobic / hydrophilic interactions, dipole-dipole interactions, ionic interactions, etc.

[0055] The solid stationary phase according to the invention advantageously comprises particles having a size between 20 and 100 microns, preferably between 30 and 80 microns.

[0056] The solid stationary phase according to the invention can be of reverse phase type or normal phase type or ion exchange type or mixed mode type. The solid stationary phase of the invention is selected according to the radiolabeled component to be purified (and therefore retained). Therefore, it can be more or less polar.

[0057] According to a preferred embodiment of the invention, the solid stationary phase is of reverse phase type. According to this embodiment, preferably, the solid stationary phase is of C8 type (phase based on silica grafted with octane chains), C18 (phase based on silica grafted with octadecyl chains) or HLB. Most preferably, the solid stationary phase is of C18 type.

[0058] According to another embodiment of the invention, the solid stationary phase is of the "mixed mode" type. A mixed mode solid stationary phase refers to a combination of a reverse phase and an ion exchange type. For example, a stationary phase called MCX ("strong cation exchange"), MAX ("strong anion exchange"), WCX ("weak cation exchange") or WAX ("weak anion exchange").

[0059] The solid stationary phase according to the present invention is captured / retained in the chamber by the housing and the chamber volume defined by the filtration device.

[0060] According to one embodiment, the solid stationary phase occupies at least 70% of the volume of the chamber (not taking into account possible microporosity of the solid stationary phase). Preferably, it occupies at least 80% of the volume of the chamber, or even at least 90% of the volume of the chamber. This makes it possible to have less "dead volume" in the kit of the invention, which is advantageous on the one hand for controlling (e.g. reducing) the injection volume, thereby improving purification by chromatography, and on the other hand for injecting the maximum amount of radiolabeled compound into the chromatography system (thereby increasing the yield).

[0061] Advantageously, one or more chemical reactions involving the radiolabeled component retained on the solid stationary phase can be carried out in the kit of the invention. Examples of chemical reactions that can be carried out are: alkaline or acidic hydrolysis, reduction, oxidation and halogenation.

[0062] According to the present invention, the test kit comprises a fluid communication device upstream and downstream of the chamber and the filtering device. In particular, the fluid communication device according to the present invention is adjacent to the filtering device. Therefore, the fluid communication device and the filtering device are arranged to allow liquid to pass through the chamber from upstream to downstream.

[0063] The fluid communication means upstream and downstream of the chamber and the filtering device according to the invention may be of the type conventionally used in the field of high pressure chromatography.

[0064] According to the invention, the housing and the fluid communication means are mechanically capable of withstanding a fluid pressure greater than or equal to 50 bar, preferably greater than or equal to 100 bar, most preferably greater than or equal to 150 bar.

[0065] According to one embodiment, the housing and / or the communication means are made of a material selected from PEEK or stainless steel or Teflon or polyoxymethylene. Preferably, the housing and the communication means are made of a material selected from PEEK or stainless steel or Teflon or polyoxymethylene (POM). More preferably, the housing and / or the communication means are made of a material selected from PEEK or stainless steel. Most preferably, the housing and the communication means are made of a material selected from PEEK or stainless steel. The materials of the housing and the communication means can be selected separately.

[0066] According to one embodiment, when the test kit housing is made of PEEK, the minimum thickness of the housing is 0.6 mm, preferably, the minimum thickness is 1 mm, more preferably, the minimum thickness is 2.5 mm. These thicknesses allow the test kit of the present invention to mechanically withstand the high pressures required by the target chromatography system, i.e., fluid pressures greater than or equal to 50 bar, greater than or equal to 100 bar, and greater than or equal to 150 bar, respectively.

[0067] According to another embodiment, when the test kit housing is made of stainless steel, the minimum thickness of the housing is 0.25 mm, preferably, the minimum thickness is 0.75 mm, more preferably, the minimum thickness is 1 mm. These thicknesses allow the test kit of the present invention to mechanically withstand the high pressure required by the target chromatography system, i.e., fluid pressures greater than or equal to 50 bar, greater than or equal to 100 bar, and greater than or equal to 150 bar, respectively.

[0068] According to another embodiment, when the test kit housing is made of Teflon, the minimum thickness of the housing is 5 mm, preferably, the minimum thickness is 8 mm, more preferably, the minimum thickness is 12 mm. These thicknesses allow the test kit of the present invention to mechanically withstand the high pressure required by the target chromatography system, i.e., fluid pressures greater than or equal to 50 bar, greater than or equal to 100 bar, and greater than or equal to 150 bar, respectively.

[0069] According to another embodiment, when the test kit housing is made of polyoxymethylene, the minimum thickness of the housing is 2 mm, preferably, the minimum thickness is 4 mm, more preferably, the minimum thickness is 5 mm. These thicknesses allow the test kit of the present invention to mechanically withstand the high pressures required by the target chromatography system, i.e., fluid pressures greater than or equal to 50 bar, greater than or equal to 100 bar, and greater than or equal to 150 bar, respectively.

[0070] In the case of a housing whose thickness varies over the width and / or length of the reagent cartridge, a minimum thickness of the housing according to the invention must be achieved over its entire width and its entire length.

[0071] According to the present invention, a device for prepurifying and injecting a liquid sample including a radiolabeled compound into a high pressure chromatography system comprises:

[0072] -Multi-way valve,

[0073] - means for supplying a liquid sample comprising a radiolabeled compound,

[0074] - elution phase pump,

[0075] - a kit according to the invention,

[0076] - discharge device,

[0077] The valve is in fluid communication with the supply, the pump, the reagent cartridge, the exhaust, and the high pressure chromatography system.

[0078] For the sake of clarity, as mentioned above, the usual injection circuit in the high pressure chromatography system is not included in the device of the present invention but is replaced by the kit, or in other words, the device of the present invention does not include an injection circuit.

[0079] According to one embodiment of the device of the present invention, the multi-way valve is a rotary valve, a solenoid valve or a pneumatic valve. The multi-way valve of the present invention can, for example, include 4-way, 6-way, 8-way or 10-way.

[0080] According to one embodiment of the device according to the invention, the means for supplying the liquid sample is an outlet of the radiosynthesis module.

[0081] According to one embodiment of the device according to the invention, the pump is in fluid communication with the elution phase reservoir.

[0082] According to one embodiment of the device according to the invention, the drain means is in fluid communication with a "waste" reservoir.

[0083] Advantageously, the device of the invention may also include means for heating the test kit, in particular the chamber of the test kit. This makes it possible to raise the temperature of the stationary phase above room temperature, which may facilitate adsorption / desorption on the stationary phase, or be useful or even necessary in the case where one or more chemical reactions are carried out in the test kit and require a temperature above room temperature to effectively occur. For example, a heating device suitable for the present invention consists of a heating sleeve surrounding the test kit.

[0084] According to one embodiment of the device according to the invention, the high pressure chromatography system is of HPLC type or SFC type.

[0085] Preferably, the chromatography system is of SFC type. The advantages of SFC type chromatography are that it allows rapid separations and that reconstitution steps can be omitted since the purified compound can be recovered in a biocompatible solvent such as ethanol.

[0086] When the chromatography system is of SFC type, the mixing chamber is arranged to be in fluid communication with (i) the multi-way valve of the apparatus of the present invention, (ii) the chromatography column and (iii) a supercritical fluid pump (eg, a CO2 pump).

[0087] It is also advantageous that, when the chromatography system is of the SFC type, the pump of the device of the invention can be used to provide a cosolvent or a modifier, which makes it possible, for example, to change the polarity of the supercritical fluid. In this case, the elution phase constitutes the cosolvent. Preferably, according to this embodiment, the elution phase is ethanol.

[0088] Alternatively, the chromatography system is of HPLC type. When the chromatography system is of HPLC type, the chromatography column is arranged to be in fluid communication with the multi-way valve of the device of the present invention.

[0089] According to the present invention, a method for prepurifying and injecting a sample including a radiolabeled compound into a high pressure chromatography system sequentially comprises a loading step and an injection step.

[0090] According to the present invention, the loading step comprises:

[0091] - providing a liquid sample comprising a radiolabeled compound to the kit according to the invention via a multi-way valve located in a loading position,

[0092] -retention of the radiolabeled compound by the kit,

[0093] - Discharge the components not retained by the reagent cartridge to a discharge device via a multi-way valve.

[0094] According to the present invention, in the loading step, the multi-way valve is located at the loading position.

[0095] According to the invention, advantageously, the liquid sample can come directly from the radiosynthesis module.

[0096] According to a preferred embodiment, the volume of the sample comprising the radiolabeled compound is between 0.5 mL and 20 mL. Preferably, the volume of the sample comprising the radiolabeled compound is between 1 mL and 15 mL, or even between 5 mL and 10 mL.

[0097] According to the present invention, during the loading step, the radiolabeled compound can be captured (e.g., by adsorption) due to the fixed solid phase, thereby achieving retention of the radiolabeled compound by the kit. Other components of the sample can be retained on the fixed phase, depending on their affinity for the phase (e.g., synthetic precursors). On the other hand, components not retained by the kit, such as synthetic solvents, are discharged to a discharge device, such as in fluid communication with a "waste" reservoir.

[0098] According to a specific embodiment of the loading step, the retention of the radiolabeled compound by the test kit can be accompanied by a chemical reaction of the radiolabeled compound in the test kit, particularly a chemical reaction on the solid stationary phase. Examples of chemical reactions that can be performed according to this embodiment are alkaline or acidic hydrolysis, reduction, oxidation and halogenation. The reaction can be performed at room temperature or at a temperature higher than room temperature. In the case where a temperature higher than room temperature is necessary or useful, a heating sleeve around the test kit, particularly its housing, can be provided.

[0099] According to the present invention, the injecting step comprises:

[0100] - pumping the elution phase towards the reagent cartridge by means of a pump through a multi-way valve in the injection position,

[0101] - releasing the radiolabeled compound retained by the kit,

[0102] - The elution phase comprising the radiolabeled compound is introduced into the high pressure chromatography system via a multi-way valve.

[0103] The release of the radiolabeled compound may be accompanied by the release of other organic compounds (synthesis residues, precursors, etc.) also retained on the kit.

[0104] According to the present invention, during the injection step, the multi-way valve is in the injection position.

[0105] According to the present invention, during the injection step, the elution phase is pumped toward the cartridge by a pump, resulting in the release (eg, by desorption) of the radiolabeled compound retained by the cartridge in the elution phase.

[0106] Said eluted phase comprising the radiolabelled compound is then introduced into a high pressure chromatography system.Preferably, the high pressure chromatography system is of HPLC type or SFC type, most preferably of SFC type, for the reasons given above.

[0107] According to a specific embodiment, the elution phase is, for example, methanol, ethanol, acetonitrile, water or an aqueous solution.Additives may also be added, for example, to influence the pH of the elution phase (eg, triethylamine, acetic acid, etc.).

[0108] When the high pressure chromatography system is of the SFC type, the elution phase is advantageously ethanol. Ethanol is a cosolvent (or modifier) ​​suitable for SFC, being a biocompatible solvent, making it possible to facilitate the reconstitution step (e.g. by reducing it to a simple dilution with water or saline or any other biocompatible buffer solution).

[0109] When the high pressure chromatography system is of the SFC type, according to an alternative embodiment, the elution phase is a supercritical phase, preferably CO 2. Advantageously, the elution phase may be a mixture of a supercritical phase (such as CO 2) and a modifier (such as ethanol).

[0110] According to the method of the invention, an elution phase comprising a radiolabeled compound is introduced into a high pressure chromatography system, said system comprising a chromatography column.

[0111] The other elements of the chromatography system, in particular of the HPLC type or SFC type, and the purification or analysis steps are known in the prior art and do not need to be described in further detail here.

[0112] According to a particular embodiment, the method of the invention comprises, after the loading step and before the injection step, at least one secondary loading step, comprising:

[0113] - providing the reagent cartridge with a secondary liquid portion comprising a radiolabeled compound via a multi-way valve located in a loading position,

[0114] -retention of the radiolabeled compound by the kit,

[0115] - Discharge the components not retained by the reagent cartridge to a discharge device via a multi-way valve.

[0116] According to this embodiment, the multiport valve is in the loading position.

[0117] The "secondary liquid fraction" refers to the fraction obtained by flushing / cleaning through the sample supply means and / or a cleaning stage of a radiosynthesis module located upstream of the supply means, or a second liquid sample comprising a radiolabeled compound (this step is a repetition of the loading step).

[0118] According to the method, the secondary liquid portion is thus further "loaded" with the cartridge, which has been loaded with the radiolabeled compound from the liquid sample during a loading step according to the invention.

[0119] In case the secondary liquid fraction comes from one or more flushing / cleaning operations, the component not retained by the cartridge and discharged to the drain consists mainly of the cleaning phase.

[0120] The device and method of the invention therefore very advantageously allow multiple flushing of the elements located upstream of pre-purification and injection (synthesis modules, supply means, etc.), thereby increasing the synthesis yield without increasing the injection volume, since the cleaning phase is eliminated / discharged (thus avoiding dilution, which is detrimental to the chromatographic performance and the purity of the final product).

[0121] According to this embodiment, the cleaning phase is selected from water, aqueous buffers and organic solvents. Preferably, the cleaning phase is water.

[0122] According to another particular embodiment, the method of the invention comprises, after the injection step, a step of regenerating the reagent cartridge. According to this embodiment, during the regeneration step, the multiway valve is in the loading position.

[0123] The regeneration step comprises in turn (i) providing the kit with a regeneration phase and (ii) discharging said phase into a discharge device. This step allows the kit to be completely cleaned, in particular any residues of the solid stationary phase, and to be restored to its initial state for a new "load-inject" cycle. This is particularly advantageous, since the kit according to the invention can thus be reused a certain number of times.

[0124] According to this embodiment, the regeneration phase is selected from water, an aqueous buffer and an organic solvent. Preferably, the regeneration phase is the elution phase.

[0125] Exemplary embodiments of the present invention are described with the aid of the accompanying drawings. It should be understood that these drawings are merely examples of how the present invention may be implemented and are in no way intended to be interpreted as limiting the scope of the present invention and the claims. In the accompanying drawings:

[0126] Figure 1 is a side cross-sectional view of a possible embodiment of a pre-purification kit according to the present invention.

[0127] Figure 2 One possible embodiment of the prepurification and injection device according to the invention is shown.

[0128] Figure 3a One possible embodiment of the device according to the invention is shown in the loading position.

[0129] Figure 3b One possible embodiment of the device according to the invention is shown in the injection position.

[0130] In the drawings, identical or similar elements have the same reference signs. Reference signs in the claims should not be construed as limiting the invention.

[0131] Figure 1 A kit for pre-purification of a sample comprising a radiolabeled compound intended for purification or analysis by high pressure chromatography is shown.

[0132] The test kit (1) comprises a housing (2) made of PEEK (a material capable of withstanding high pressures, such as in the case of SFC or HPLC type chromatography) and having a thickness of 4.5 mm.

[0133] The kit (1) further comprises a filtering device (3).

[0134] The reagent kit (1) comprises a chamber (4), the volume of which is defined by a housing (2) and a filtering device (3).

[0135] The test kit (1) also includes PEEK fluid communication devices (5) located upstream and downstream of the chamber (4) and the filtering device (3). These communication devices (5) make it possible to connect the test kit (1) fluidically to a device (such as the device according to the present invention described below) into which it is integrated. The fluid communication device (5) is adjacent to the filtering device. The fluid communication device (5) and the filtering device (3) allow the liquid sample to pass through the chamber from upstream to downstream.

[0136] The chamber (4) comprises a reverse phase C18 type solid stationary phase for retaining the radiolabeled compound (e.g. by adsorption). The solid stationary phase thus advantageously allows a pre-purification of the sample. The filtering means (3) make it possible (together with the housing) to retain the solid stationary phase in the volume of the chamber while allowing the liquid sample to pass by filtering potential solid particles (thus advantageously preventing any clogging of the column during subsequent purification by high pressure chromatography).

[0137] Figure 2 A device (6) for the pre-purification and injection of a liquid sample comprising a radiolabeled compound in a high pressure chromatography system (7) is shown. The device (6) comprises a multi-way valve (8). The multi-way valve (8) makes it possible to fluidically connect (12) all elements of the device together, in particular a supply device (9), an elution phase pump (10), a reagent kit (1), and an exhaust device (11). The valve also makes it possible to fluidically connect (12) the high pressure chromatography system (7) to the device of the invention. The supply device (9) makes it possible to supply the device (6) with a liquid sample of a radiolabeled compound (e.g. from a radiosynthesis module). The elution phase pump (10) is used to pump the elution phase in the device (6). The reagent kit (1) allows the pre-purification of a liquid sample comprising a radiolabeled compound.

[0138] Figure 3a and 3b Shows the operation Figure 2 Device (6) and implementation scheme of the method of the present invention.

[0139] Figure 3a The loading step is shown. In this first step, the multi-way valve (6) (8) is in the loading position (13). The loading position (13) makes it possible to supply a liquid sample including a radiolabeled compound to the test kit (1) via the supply device (9). As the sample passes through the test kit (1), the radiolabeled compound is retained by the solid stationary phase. Therefore, this advantageously makes it possible to concentrate and / or pre-purify the radiolabeled compound at this stage of the process. The components (14) not retained by the test kit (1) (i.e., components with little or no affinity for the stationary phase) flow out of the test kit and reach the discharge device (11) through the multi-way valve (8).

[0140] The loading position can also be used to perform a secondary loading step by supplying a secondary liquid portion (e.g., a portion rinsed / cleaned with a cleaning stage of a radioactive synthesis module located upstream of the supply device (9)) to the test kit (1) via a supply device (9). In this case, the secondary liquid portion is therefore further "loaded" with the test kit, which has already been loaded with the radioactive labeled compound of the liquid sample supplied in the first loading step. In this secondary loading step (which can be repeated), the radioactive labeled compound from, for example, a cleaning synthesis module is therefore recovered on the test kit, which will help to improve the radioactive synthesis yield. On the other hand, the cleaning phase and other components not retained on the test kit will be moved to the discharge device (11). Therefore, this allows the cleaning of the radioactive synthesis module and the supply device to recover as much radioactive labeled compound as possible without affecting the final volume of the sample injected into the chromatographic system (7). In fact, this volume can be selected / controlled in the next injection step.

[0141] Furthermore, during this loading step, the elution phase (15) can be fed directly to the chromatography system (7) via a multi-way valve (8) in a conventional manner. This allows the chromatography column to be equilibrated during the loading of the reagent cartridge (1).

[0142] Figure 3bThe injection step is shown. In the second step of the process, the multi-way valve (8) is in the injection position (16). The injection position (16) makes it possible to elute the kit (1) of the radiolabeled compound retained therein and inject the concentrated and / or pre-purified sample into the chromatographic system (7) for purification (or analysis). This also makes it possible to filter the sample through the kit (1) in advance and thus prevent clogging of the chromatographic column. In this injection step, the pump (10) pumps the elution phase (15) into the kit (1) through the multi-way valve (8), which releases the radiolabeled compound retained by the kit (1) and thus elutes it. The elution phase comprising the radiolabeled compound (17) is then injected into the high pressure chromatography system (7) through the multi-way valve (8). Therefore, compared with the initial sample (before passing through the kit), the elution phase comprising the radiolabeled compound (17) constitutes a concentrated and / or pre-purified sample. In particular, the initial volume can be adjusted (e.g., reduced), the solvent can be changed, certain components (components not retained by the kit) can be removed, and any solid particles present can be filtered.

[0143] Furthermore, during this injection step, the supply device (9) is fluidically connected to the discharge device (11). This makes it possible to clean the supply device (9) and flush it before the next loading phase. This has the advantage of reducing the risk of subsequent contamination and / or the risk of forming aggregates that could clog the system, which needs to be maintained.

[0144] It should be understood that the present invention is by no means limited to the above-described embodiments and can be modified without departing from the scope of the claims. It should also be understood that the present invention also comprises all possible combinations of features and preferred features described herein and mentioned in the claims.

[0145] Furthermore, the following examples are provided for illustrative purposes and are not intended to limit the scope of the present invention. Example

[0146] This example corresponds to the automated purification of a radioactive tracer labeled with fluorine-18, namely compound [ 18 F]-AV-45.

[0147] The invention is made of PEEK and has a volume of 785 mm 2 The kit was manually filled with 90 mg of HLB type stationary phase with a particle size of 50-65 microns (hydrophilic-lipophilic balance, Waters HLB Plus Short kit, reference number 186000132).

[0148] Automated radiosynthesis (capture of fluorine-18 on the QMA kit, elution of fluorine-18, drying of fluorine-18 in the reactor, labeling, hydrolysis, neutralization) in the first module ( xSeed™) with 2.0 mg of precursor in 2.0 mL of DMSO.

[0149] The neutralized reaction material was manually diluted to 10 mL with water using a 10 mL luer-lock syringe. It was manually placed in the syringe pump and then automatically loaded with the kit according to the present invention. The speed of the syringe pump was 4 mm / 6 seconds and a relative vacuum of -0.9 bar was applied. After loading was completed, the operator did not rinse the syringe by The software is in injection mode (according to Figure 2 ) activates the injection valve (8).

[0150] The UV trajectory was recorded by manually starting the acquisition simultaneously through the SFC module software.

[0151] To monitor the fluorine-18 labeled product, one radioactivity detector is placed in contact with the cartridge and a second detector is placed at the outlet of the SFC purification system. The second radioactivity detector allows the operator to actuate the collection valve so that the radioactivity peak is collected in the collection vessel.

[0152] For this example, the radioactivity in the 10 ml syringe prior to loading onto the cartridge was 11.4 mCi (420 Mbq).

[0153] The "semi-preparative" column used was a Phenomenex Polar RP 10 μm, 250 x 10 mm column.

[0154] The conditions for SFC are as follows:

[0155] - Eluent: CO2 / EtOH 95 / 5;

[0156] - Flow rate: 20 mL / min;

[0157] - Pressure regulator at 150 bar, heated to 60°C;

[0158] -UV detection wavelength 254nm;

[0159] - Temperature of SFC system: between 25°C and 30°C.

[0160] According to the measurements performed, 90% of the activity on the kit was eluted within 1 minute with the eluent CO2 / EtOH 95 / 5.

[0161] The radioactive peak associated with compound [18F]-AV-45 appeared between 6'20" and 7'30". For comparison, in the HPLC purification [ 18 F]-AV-45 usually elutes around 15'.

[0162] At the end of the collection, the final container is manually unscrewed from the collection module and inserted into the activity meter to measure its activity.

[0163] The following table gives the results of this example (residual activity during the process and synthesis yield measurements (A0: initial activity; dc: "decay correction", correction for the decay of fluorine-18).

[0164]

[0165]

[0166] * At the end of the synthesis, t0+35 minutes, the activity was 4.57 mCi, i.e. a decay-uncorrected yield of 33%.

[0167] **4% activity of liquid waste associated with kit loading is labeled product.

Claims

1. A kit (1) for prepurifying a liquid sample comprising a radiolabeled compound to be purified or analyzed by high pressure chromatography, the kit (1) comprising: - a housing (2), - a filtering device (3), - a chamber (4) whose volume is defined by the housing (2) and the filtering device (3), the chamber comprising a solid stationary phase, and - fluid communication means (5) located upstream and downstream of the chamber (3) and the filtering means (3), The housing (2) and the fluid communication device (5) are mechanically capable of withstanding a fluid pressure greater than or equal to 50 bar.

2. The kit according to claim 1, characterized in that The housing (2) and the communication device (5) are capable of mechanically withstanding a fluid pressure greater than or equal to 100 bar.

3. The kit according to claim 1 and 2, characterized in that The housing (2) and / or the connecting device (5) are made of a material selected from PEEK or stainless steel.

4. The kit according to any one of the preceding claims, characterized in that The solid stationary phase is of reverse phase type.

5. The kit according to any one of the preceding claims, characterized in that The volume of the chamber (4) is 50 mm 3 Up to 2000mm 3 between.

6. A device (6) for prepurifying and injecting a liquid sample comprising a radiolabeled compound into a high pressure chromatography system (7), the device comprising: -Multi-way valve (8), - means (9) for supplying a liquid sample comprising a radiolabeled compound, - an elution phase pump (10), - a kit (1) according to claims 1 to 5, - a discharge device (11), The valve is in fluid communication (12) with the supply device (9), the pump (10), the reagent cartridge (1), the discharge device (11) and the high pressure chromatography system (7).

7. Device according to the preceding claim, characterized in that The chromatography system (7) is of HPLC type or SFC type.

8. Device according to the preceding claim, characterized in that The chromatography system (7) is of SFC type.

9. A method for prepurifying and injecting a liquid sample comprising a radiolabeled compound into a high pressure chromatography system (7), said method using a device according to any one of claims 6 to 8.

10. A method for prepurifying a liquid sample comprising a radiolabeled compound and injecting it into a high pressure chromatography system (7), the method comprising: (i) a loading step, comprising: - supplying a liquid sample comprising a radiolabeled compound to a reagent kit (1) according to claims 1 to 5 via a multi-way valve (8) in a loading position (13), - retaining the radiolabeled compound by the kit (1), - discharging the components (14) not retained by the reagent kit (1) to the discharge device (11) through the multi-way valve (8); (ii) an injection step, comprising: - pumping the elution phase (15) by means of a pump (10) through a multi-way valve (8) in the injection position (16) towards the reagent cartridge (1), - releasing the radiolabeled compound retained by the kit (1), - The elution phase (17) comprising the radiolabeled compound is introduced into the high pressure chromatography system (7) via the multi-way valve (8).

11. The method according to the preceding claim, characterized in that The high pressure chromatography system (7) is of HPLC type or SFC type.

12. The method according to the preceding claim, characterized in that The high pressure chromatography system (7) is of SFC type.

13. The method according to the preceding claim, characterized in that The elution phase (15) is ethanol.

14. The method according to any one of claims 10 to 13, characterized in that After the injection step, the method comprises a step of regenerating the kit (1), the steps comprising, in order: - supplying the regeneration phase via a multi-way valve (8) located in the loading position (13) of the reagent cartridge (1), and - Discharge of the phase to a discharge device (11) via the multi-way valve (8).

15. The method according to the preceding claim, characterized in that The regeneration phase is selected from water, aqueous buffer and organic solvent.

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