Cartridge, pre-purification and injection device, and pre-purification and injection method for pre-purifying a radiolabeled compound sample for purification by high pressure chromatography - Patents.com

Abstract

The present invention relates to a cartridge for the pre-purification of liquid samples containing radiolabeled compounds intended for purification by high pressure chromatography, comprising a casing (2), a filtering means (3), a chamber (4) containing a solid stationary phase and a fluid communication means (5), the casing (2) and the fluid communication means (5) being mechanically capable of withstanding fluid pressures of more than 50 bar. The present invention also relates to a pre-purification and injection device incorporating the cartridge, as well as a pre-purification and injection method using the cartridge.

Description

[Technical field]

[0001] The present invention relates to the field of synthesis and purification of radiolabeled compounds, e.g. as injectable radiotracers for positron emission tomography (PET) analysis. In particular, the present invention relates to a cartridge for prepurifying a liquid feed sample resulting from the synthesis of radiolabeled compounds for analysis / separation / purification 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. [Background technology]

[0002] The synthesis (or radiosynthesis) of radiolabeled compounds is generally based on "wet" chemistry, where a radioactive synthon is reacted with a non-radioactive precursor in solution in a synthesis module or radiosynthesizer. Since the radioactive compound is intended to be injected into the patient's body, this approach necessarily requires steps that involve purification of the synthesis (raw sample containing the radiolabeled compound).

[0003] A well-known and commonly used method for purifying biological samples from radioactive synthesis is high performance or high pressure liquid chromatography (HPLC). This purification step must be followed by a re-purification step with a biocompatible solvent, since the usual HPLC solvents (such as acetonitrile) are not suitable for injection into the human or animal body.

[0004] When purifying biological samples from the synthesis of radiolabeled compounds, the use of supercritical fluid chromatography (or SFC) is also known, but to a lesser extent, and is mainly used for the purification of carbon-11 labeled compounds or compounds with low levels of radioactivity. This method uses a supercritical fluid (commonly carbon dioxide) as the mobile phase, usually in combination with a co-solvent or "modifier" that changes the polarity (e.g., ethanol). SFC can speed up the separation step compared to HPLC (at least 2-3 times), which is a major advantage in the case of short-lived radioisotopes, since it improves the final synthesis yield. Moreover, with SFC, the purified sample does not need to be reformulated, since after simple dilution (e.g., saline or aqueous buffer), it can be recovered in ethanol, which is suitable for injection into the body.

[0005] In any of these purification methods, the raw sample from the radioactive synthesis is generally injected into the chromatographic column by an injection system consisting of an injection loop and a valve. The loop allows the injection into the column of a maximum volume equal to the volume of the loop (allowing reproducibility of the injection volume when the full volume of the loop is used), and the valve allows the fluidic connection of the different elements (the "outlet" of the synthesizer, the loop, the pump, the "waste" container), etc. The disadvantage of such a loop is that the delivered volume is at most limited to the volume of the loop, unless the loop is filled and injected (automatically) several times, which is very time-consuming and should be absolutely avoided in the case of radioactive synthesis. Moreover, in this case there is always a risk of loss of yield, for example by overflow.

[0006] In particular, HPLC methods can be used to manage high injection volumes (e.g. up to 10 ml), thus allowing constant speed since the loop is loaded only once, and also limiting yield losses (due to loss of activity in the radiosynthesizer) since one or more rinse steps can be performed to maximize the recovery of biosamples from the synthesis of radiolabeled compounds. On the other hand, the dilution that occurs 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 (1 ml or less) because larger injection volumes result in severe decompression when the injection valve is reset to the load position. Therefore, the use of this separation method is not suitable for the synthesis of large volumes, i.e., volumes greater than 1 ml.

[0008] However, conventional radiosynthesizer configurations do not allow automatic transfer of volumes less than 1 ml without loss of radioactivity. For raw sample volumes greater than 1 ml, (pre)concentration, for example by evaporation of the solvent, is conceivable in order to use SFC and enjoy the aforementioned advantages, but injection of small amounts into the loop must be done manually to limit losses, which is not conceivable for the amounts of radioactivity used in the production. Moreover, for example in the case of short-lived radioisotopes, the addition of an evaporation step automatically affects the synthesis yield.

[0009] More specifically, although there are separation systems using SFC (industrial fractionation and filtration) with larger injection loops (e.g., 5 ml), these systems are bulky and very expensive, and therefore have rarely been applied to the synthesis of radiolabeled compounds.

[0010] Furthermore, due to the risk of encountering problems with clogging of the chromatographic columns, the raw samples obtained from the radioactive synthesis often need to be filtered first, again an additional step that extends the time of the complete synthesis-purification process. Moreover, this filtration step requires a strict selection of filters to minimize losses at the filter membrane (e.g. due to chemical interactions). One object of the present invention is to address the drawbacks of the prior art mentioned above. Summary of the Invention [Problem to be solved by the invention]

[0011] In particular, one object of the present invention is to provide an apparatus and method for pre-purifying and injecting into a high-pressure chromatography system a raw sample resulting from the synthesis of a radiolabeled compound, which reduces the time required to obtain a purified radiolabeled compound compared to prior art systems, thus allowing access to a higher final yield.

[0012] It is also an object of the present invention to provide an apparatus and method for pre-purifying and injecting a biosample resulting from the synthesis of a radiolabeled compound into a high pressure chromatography system, regardless of the initial volume of the biosample.

[0013] Another object of the present invention is to provide an apparatus and method for pre-purifying and automatically injecting raw samples resulting from the synthesis of radiolabeled compounds into a high pressure chromatography system without manual manipulation.

[0014] Another object of the present invention is to provide an apparatus and a method for the pre-purification and injection of a biosample resulting from the synthesis of a radiolabeled compound, which allows one or more washings of the radiosynthesizer without increasing the volume injected into the column (thus optimizing the yield of the radiosynthesis by minimizing losses in the radiosynthesizer).

[0015] Another object of the present invention is to provide an apparatus and method for pre-purifying and injecting into a high pressure chromatography system a biosample resulting from the synthesis of a radiolabeled compound, which allows the injection volume to be controlled independently of the initial amount of biosample, thus allowing better chromatographic reproducibility.

[0016] Yet another object of the present invention is to provide an apparatus and a method for the pre-purification and injection of a raw sample from the synthesis of a radiolabeled compound, which allows a single (automatic) injection into a chromatography system, regardless of the initial amount of the sample, thus saving time. Another object of the present invention is to provide an apparatus and a method for the pre-purification and injection of a raw sample from the synthesis of a radiolabeled compound, which also allows filtration, thus preventing subsequent clogging of the chromatography column. [Means for solving the problem]

[0017] To achieve these aims, the invention provides a cartridge for the pre-purification of a liquid sample containing a radiolabelled compound intended to be purified or analysed by high pressure chromatography.

[0018] In particular, the cartridge of the present invention comprises a casing, a filtering means, a chamber whose volume is defined by the casing and the filtering means, the chamber containing a solid stationary phase, and fluid communication means upstream and downstream of the chamber and the filtering means, the casing and the fluid communication means being mechanically capable of withstanding a fluid pressure of 50 bar or more.

[0019] The present invention also provides an apparatus for pre-purifying and injecting a liquid sample containing a radiolabeled compound into a high pressure chromatography system.

[0020] In particular, the device of the present invention comprises a multi-way valve, a supply means for a liquid sample containing a radiolabeled compound, an elution phase pump, a cartridge according to the present invention, and an exhaust means, the valve being in fluid communication with the supply means, the pump, the cartridge, the exhaust means, and the high pressure chromatography system.

[0021] Finally, the present invention also provides a method for pre-purifying a liquid sample containing a radiolabeled compound and injecting it into a high pressure chromatography system.

[0022] In particular, the method of the present invention comprises (i) a loading step and (ii) an injection step, (i) the loading step comprising the steps of providing a cartridge according to the present invention with a liquid sample comprising a radiolabelled compound through a multi-way valve in a loading position, retaining the radiolabelled compound by the cartridge and removing through the multi-way valve to an outlet means elements not retained by the cartridge, and (ii) the injection step comprising the steps of aspirating the elution phase by a pump, releasing the radiolabelled compound retained by the cartridge and introducing the elution phase comprising the radiolabelled compound through the multi-way valve into the high pressure chromatography system.

[0023] The present invention is therefore based on a novel and original approach. In particular, the inventors have surprisingly found that by replacing the injection loop in a conventional injection device of a high pressure chromatography system (of the HPLC or SFC type) with a cartridge of the characteristics of the present invention, it is possible to (i) significantly shorten the separation time (up to 4 times) and (ii) significantly improve the synthesis yield by allowing cleaning of the radiosynthesizer (thus minimizing the loss of compounds in the radiosynthesizer) without increasing the injection volume into the column, regardless of the initial volume of the sample containing the radiolabeled compound. Therefore, for the sake of clarity, it is understood that the conventional injection loop in a high pressure chromatography system is not included in the device of the present invention, in other words, the device of the present invention does not include an injection loop.

[0024] Furthermore, the present invention also makes it possible to automate the injection, avoiding manual procedures.

[0025] Further features, details and advantages of the invention are contained in the following description and figures, without being limiting.

[0026] In the present specification and claims, it is clearly understood that the term "one" as used herein means "at least one" and should not be limited to "a single one" unless expressly indicated otherwise. The terms "comprise", "have", "include" and "have" have an open meaning and do not exclude the presence of additional elements. Furthermore, when a range of values ​​is given, the lowest and highest numerical values ​​are included. Finally, all integral and subdomain values ​​within a numerical range are expressly included as if expressly stated.

[0027] According to the invention, the cartridge allows the pre-purification of a liquid sample containing a radiolabelled compound intended to be purified or analysed by high pressure chromatography.

[0028] According to the invention, the pre-purification of the liquid sample may also correspond to or include the concentration of the sample, in particular the concentration of the radiolabeled compound.

[0029] In particular, a sample according to the invention may consist of one radiolabeled compound or several radiolabeled compounds, for example (i) organic molecules such as sugars (e.g. glucose) or amino acids, bearing radioisotopes such as carbon-11, fluorine-18, iodine-131, yttrium-90 or actinium-225.

[0030] According to the invention, the liquid sample consists of radiolabeled compounds intended to be purified or analyzed by high pressure chromatography. The cartridge of the invention is in fact suitable for analytical chromatography (making it possible, for example, to detect the presence or production of one or more compounds) or for preparative or semi-preparative chromatography (making it possible, for example, to take one or more purified compounds from a raw sample obtained from a synthesis).

[0031] According to the invention, the liquid sample may be a sample in an aqueous phase, a sample in an organic phase, a sample in two phases (aqueous phase / organic phase) or an emulsion. Preferably, the liquid sample is an aqueous single-phase sample. The aqueous phase according to the invention consists of at least 40% (by volume) water, preferably at least 50% water, or even at least 70% water.

[0032] According to the invention, the cartridge comprises a filtering means which allows the passage of fluids and allows (by the casing) the retention / capture of the solid stationary phase within the chamber by filtering potential solid particles from the raw sample (thus preventing clogging of the chromatographic column during purification by high pressure chromatography).

[0033] Preferably, the filtration means is selected from a filtration membrane and a frit.

[0034] According to the present invention, the cartridge includes a chamber whose volume is defined by a casing and a filtering means. 3 Most preferably, the chamber has a volume of 50 to 1000 mm. 3 These volumes are particularly suitable for the volumes of liquid samples obtained from radioactive synthesis.

[0035] According to the invention, the chamber consists of a solid stationary phase. The term "solid stationary phase" refers to the generally accepted definition in the field of separation / chromatography of chemical components in a mixture, where "solid stationary phase" refers to a compound in solid form used to hold / adsorb the components of the mixture to be separated. This phase is immobile in contrast to the mobile phase (liquid or gas) which moves through the stationary phase. There are various types of adsorption interactions that occur between the solid stationary phase and the components, such as hydrogen bonding, hydrophobic / hydrophilic interactions, dipole-dipole interactions, ionic interactions, etc.

[0036] The solid stationary phase according to the invention advantageously consists of particles with a size ranging from 20 to 100 microns, preferably ranging from 30 to 80 microns.

[0037] The solid stationary phase according to the invention may be of reversed phase, normal phase, ion exchange or mixed mode type. The solid stationary phase according to the invention is selected according to the radiolabeled component to be purified (and therefore retained). It may therefore be highly or less polar.

[0038] According to a preferred embodiment of the present invention, the solid stationary phase is of the reversed phase type. According to this embodiment, preferably, the solid stationary phase is of the C8 type (a silica-based phase grafted with octane chains), C18 type (a silica-based phase grafted with octadecyl chains) or HLB. Most preferably, the solid stationary phase is of the C18 type.

[0039] According to another embodiment of the invention, the solid stationary phase is of the "mixed mode" type. A mixed mode type solid stationary phase refers to a combination of a reversed phase type and an ion exchange type, such as the stationary phases called MCX ("strong cation exchange"), MAX ("strong anion exchange"), WCX ("weak cation exchange") or WAX ("weak anion exchange").

[0040] The solid stationary phase according to the present invention is captured / retained within the chamber by a filtration means which defines the volume of the casing and the chamber.

[0041] According to one embodiment, the solid stationary phase occupies at least 70% of the chamber volume (not taking into account possible microscopic porosity of the solid stationary phase). Preferably, it occupies at least 80% or even at least 90% of the chamber volume. This allows the cartridge of the invention to have a low "dead volume", which is advantageous, on the one hand, for controlling (for example reducing) the injection volume and thus improving the chromatographic purification, and, on the other hand, allows the maximum injection of the radiolabeled compound into the chromatographic system (thus increasing the yield).

[0042] Advantageously, one or more chemical reactions involving the radiolabeled components retained on the solid stationary phase can be carried out in the cartridge of the invention. Examples of chemical reactions that can be carried out are basic or acidic hydrolysis, reduction, oxidation and halogenation.

[0043] According to the invention, the cartridge comprises fluid communication means upstream and downstream of the chamber and the filtering means. In particular, the fluid communication means according to the invention is adjacent to the filtering means. In this way, the fluid communication means and the filtering means are arranged to allow liquid to pass from upstream to downstream through the chamber.

[0044] According to the invention, the cartridge comprises fluid communication means upstream and downstream of the chamber and the filtering means. In particular, the fluid communication means according to the invention is adjacent to the filtering means. In this way, the fluid communication means and the filtering means are arranged to allow liquid to pass from upstream to downstream through the chamber.

[0045] According to the invention, the casing and the fluid communication means are mechanically capable of withstanding fluid pressures of at least 50 bar, preferably at least 100 bar, most preferably at least 150 bar.

[0046] According to one embodiment, the casing and / or the communication means are made of a material selected from PEEK or stainless steel or Teflon or polyoxymethylene. Preferably, the casing and the communication means are made of a material selected from PEEK or stainless steel or Teflon or polyoxymethylene (POM). More preferably, the casing and / or the communication means are made of a material selected from PEEK or stainless steel. Most preferably, the casing and the communication means are made of a material selected from PEEK or stainless steel. The materials of the casing and the communication means can be selected separately.

[0047] According to one embodiment, when the cartridge casing is made of PEEK, said casing has a minimum thickness of 0.6 mm, preferably a minimum thickness of 1 mm, more preferably a minimum thickness of 2.5 mm, which allows the cartridge of the invention to mechanically withstand the high fluid pressures required by the chromatography systems of interest, i.e., 50 bar or more, 100 bar or more, 150 bar or more, respectively.

[0048] According to another embodiment, when the cartridge casing is made of stainless steel, said casing has a minimum thickness of 0.25 mm, preferably a minimum thickness of 0.75 mm, more preferably a minimum thickness of 1 mm, which allows the cartridge of the invention to mechanically withstand the high pressures required by the chromatography systems of interest, i.e., fluid pressures of 50 bar or more, 100 bar or more, and 150 bar or more, respectively.

[0049] According to another embodiment, when the cartridge casing is made of Teflon, said casing has a minimum thickness of 5 mm, preferably a minimum thickness of 8 mm, more preferably a minimum thickness of 12 mm, which allows the cartridge of the invention to mechanically withstand the high pressures required by the chromatography systems of interest, i.e., fluid pressures of 50 bar or more, 100 bar or more, and 150 bar or more, respectively.

[0050] According to another embodiment, when the cartridge casing is made of polyoxymethylene, said casing has a minimum thickness of 2 mm, preferably a minimum thickness of 4 mm, more preferably a minimum thickness of 5 mm, which allows the cartridge of the invention to mechanically withstand the high pressures required by the chromatography systems of interest, i.e. fluid pressures of 50 bar or more, 100 bar or more, 150 bar or more, respectively.

[0051] If the casing has a thickness that varies across the width and / or length of the cartridge, the minimum thickness of the casing according to the invention must be achieved along its entire width and length.

[0052] According to the present invention, an apparatus for pre-purifying a liquid sample containing a radiolabeled compound and injecting it into a high-pressure chromatography system comprises a multi-way valve, a supply means for the liquid sample containing the radiolabeled compound, an elution phase pump, a cartridge according to the present invention, and an ejection means, the valve being in fluid communication with the supply means, the pump, the cartridge, the ejection means and the high-pressure chromatography system.

[0053] For clarity, and as already explained above, the usual injection loop in a high pressure chromatography system is not included in the device of the present invention, but is replaced by a cartridge.

[0054] 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 be, for example, a 4-way valve, a 6-way valve, an 8-way valve or a 10-way valve.

[0055] According to one embodiment of the device of the invention, the liquid sample supply means is the outlet of a radiosynthesis module.

[0056] According to one embodiment of the device of the present invention, the pump is in fluid communication with the elution phase reservoir.

[0057] According to one embodiment of the device of the present invention, the drainage means is in fluid communication with a "waste" reservoir.

[0058] Advantageously, the device of the invention may also comprise means for heating the cartridge, in particular the chamber of the cartridge. This allows the temperature of the stationary phase to be raised above ambient temperature, which may aid in adsorption / desorption onto the stationary phase, or may be beneficial or even necessary if one or more chemical reactions are carried out within the cartridge and require a temperature higher than ambient to take place effectively. For example, a heating means compatible with the invention may consist of a heating sleeve surrounding the cartridge.

[0059] Advantageously, the device of the invention may also comprise means for heating the cartridge, in particular the chamber of the cartridge. This allows the temperature of the stationary phase to be raised above ambient temperature, which may aid in adsorption / desorption onto the stationary phase, or may be beneficial or even necessary if one or more chemical reactions are carried out within the cartridge and require a temperature higher than ambient to take place effectively. For example, a heating means compatible with the invention may consist of a heating sleeve surrounding the cartridge.

[0060] Preferably, the chromatographic system is of the SFC type, which has the advantage that it allows rapid separation and allows the purified compounds to be recovered in a biocompatible solvent (e.g. ethanol), thus making it possible to avoid reworking steps.

[0061] When the chromatography system is of the SFC type, the mixing chamber is provided 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, such as a CO2 pump.

[0062] Also advantageously, when the chromatographic system is of the SFC type, the pump of the device of the invention can be used to supply a co-solvent or modifier making it possible, for example, to modify the polarity of the supercritical fluid. In this case, the elution phase constitutes said co-solvent. Preferably, according to this embodiment, the elution phase is ethanol.

[0063] Alternatively, the chromatography system is of the HPLC type. When the chromatography system is of the HPLC type, a chromatography column is placed in fluid communication with the multi-way valve of the device of the invention.

[0064] According to the present invention, a method for pre-purification and injection of a sample containing a radiolabeled compound into a high pressure chromatography system comprises, in sequence, a loading step and an injection step.

[0065] According to the present invention, the loading step includes the steps of supplying a liquid sample containing a radiolabeled compound to a cartridge according to the present invention through a multi-way valve in a loading position, retaining the radiolabeled compound by the cartridge, and discharging elements not retained in the cartridge to a discharge means through the multi-way valve.

[0066] According to the invention, in the loading step the multi-way valve is in the loading position.

[0067] According to the invention, advantageously, the liquid sample is provided directly from the radiosynthesis module.

[0068] According to a preferred embodiment, the amount of the sample containing the radiolabeled compound is between 0.5 ml and 20 ml. Preferably, the volume of the sample consisting of the radiolabeled compound is between 1 ml and 15 ml, even more preferably between 5 ml and 10 ml.

[0069] According to the invention, in the loading step, retention of the radiolabeled compounds by the cartridge is achieved thanks to a solid stationary phase that allows in particular to capture the radiolabeled compounds (e.g. by adsorption). Other components of the sample can be retained on the stationary phase by their affinity for the stationary phase (e.g. synthesis precursors). Components not retained on the cartridge, e.g. synthesis solvents, are discharged into the discharge means.

[0070] According to a particular embodiment of the loading step, the retention of the radiolabeled compound by the cartridge can involve a chemical reaction of the radiolabeled compound in the cartridge, in particular on the solid stationary phase. Examples of chemical reactions that can be carried out according to this embodiment are basic or acid hydrolysis, reduction, oxidation and halogenation. The reaction can be carried out at ambient temperature or above ambient temperature. In cases where temperatures above ambient temperature are necessary or useful, a heating sleeve can be provided surrounding the cartridge, in particular its casing.

[0071] According to the present invention, the injection step includes the steps of pumping the elution phase towards the cartridge through a multi-way valve in the injection position, releasing the radiolabeled compound retained in the cartridge, and introducing the elution phase containing the radiolabeled compound through the multi-way valve into the high pressure chromatography system.

[0072] The release of the radiolabeled compound may be accompanied by the release of other organic compounds (synthetic residues, precursors, etc.) retained in the cartridge.

[0073] According to the invention, in the injecting step the multi-way valve is in the injecting position.

[0074] According to the invention, in the injection step, an elution phase is drawn towards the cartridge by a pump and the radiolabelled compound retained in the cartridge is released (eg by desorption) in the elution phase.

[0075] The elution phase containing the radiolabeled compound is then introduced into a high pressure chromatography system, preferably of the HPLC or SFC type, most preferably of the SFC type for the reasons mentioned above.

[0076] According to certain embodiments, the elution phase is, for example, methanol, ethanol, acetonitrile, water or an aqueous solution. Also, additives may be added, for example to affect the pH of the elution phase (for example, triethylamine, acetic acid, etc.).

[0077] When the high pressure chromatography system is of the SFC type, the elution phase is advantageously ethanol, which is a suitable co-solvent (or modifier) ​​for SFC and is a biocompatible solvent, making it possible to facilitate the modification step (for example by simple dilution with water or saline or other biocompatible buffer solutions).

[0078] According to another embodiment, when the high pressure chromatography system is of the SFC type, the elution phase is a supercritical phase, preferably CO2. Advantageously, the elution phase may be a mixture of a supercritical phase, for example CO2, and a modifier, for example ethanol.

[0079] According to the method of the present invention, an elution phase containing the radiolabeled compound is introduced into a high pressure chromatography system, the system including a chromatography column.

[0080] The other elements of the chromatography systems, in particular those of the HPLC or SFC type, and the purification or analysis steps are known from the prior art and need not be detailed here further.

[0081] According to a particular embodiment, the method of the invention comprises at least one secondary loading step after the loading step and before the injection step, the secondary loading step comprising the steps of: supplying a secondary liquid fraction comprising the radiolabeled compound to the cartridge through the multi-way valve in the loading position, retaining the radiolabeled compound in the cartridge, and discharging elements not retained in the cartridge through the multi-way valve to the discharge means.

[0082] According to this embodiment, the multiport valve is in the load position.

[0083] "Secondary liquid fraction" refers to the fraction resulting from the rinsing / washing by a washing step of the sample supply means and / or the radiosynthesis module located upstream of the supply means, or a secondary liquid sample containing radiolabeled compound (this step is then a repeat of the loading step).

[0084] According to this method, the second liquid fraction thus further "loads" the cartridge that has already been loaded with radiolabelled compound from the liquid sample in a loading step according to the invention.

[0085] Where the secondary liquid fraction is derived from one or more rinse / wash operations, the components not retained in the cartridge and discharged into the discharge means will consist primarily of the wash phase.

[0086] The device and method of the invention therefore very advantageously allow several washings of the elements located upstream of the pre-purification and injection (synthesis module, supply means, etc.), thus increasing the synthesis yield without increasing the injection volume, since the washing phase is removed / discharged (thus avoiding dilution detrimental to the chromatographic performance and purity of the final product).

[0087] According to this embodiment, the washing phase is selected from water, an aqueous buffer and an organic solvent. Preferably, the washing phase is water.

[0088] According to another particular embodiment, the method of the invention comprises, after the injection step, a cartridge regeneration step, wherein in the regeneration step the multiport valve is in the loading position.

[0089] The regeneration step comprises, in turn, (i) the step of supplying the cartridge with a regeneration phase and (ii) the step of discharging the phase into a discharging means. This step makes it possible to thoroughly wash the cartridge, and in particular the solid stationary phase, from residues and to return it to an initial state to allow new "loading-injecting" cycles. This is particularly advantageous since the cartridge of the invention can be reused a certain number of times.

[0090] According to this embodiment, the regeneration phase is selected from water, an aqueous buffer, and an organic solvent. Preferably, the regeneration phase is an elution phase.

[0091] Exemplary embodiments of the invention are illustrated in the figures, it being understood that these figures are merely examples of how the invention may be practiced, and are not intended to be construed in any way as limiting the scope of the invention and claims. [Brief description of the drawings]

[0092] [Figure 1] FIG. 1 is a side cross-sectional view of a possible embodiment of a pre-purification cartridge according to the present invention. [Diagram 2] FIG. 2 shows a possible embodiment of a pre-purification and injection device according to the invention. [Figure 3a] FIG. 3a shows a possible embodiment of the device according to the invention in the loading position. [Figure 3b] FIG. 3b shows a possible embodiment of the device according to the invention in the injection position. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0093] In the drawings, the same reference numbers refer to the same or similar elements. Reference numbers in the claims should not be construed as limiting the invention.

[0094] FIG. 1 shows a cartridge (1) for the pre-purification of samples containing radiolabeled compounds intended for purification or analysis by high pressure chromatography.

[0095] The cartridge (1) comprises a casing (2) made of PEEK (a material capable of withstanding high pressures such as those in SFC or HPLC type chromatography) and having a thickness of 4.5 mm.

[0096] The cartridge (1) also comprises a filtering means (3).

[0097] The cartridge (1) comprises a chamber (4), the volume of which is defined by a casing (2) and a filtering means (3).

[0098] The cartridge (1) also comprises fluid communication means (5) made of PEEK, upstream and downstream of the chamber (4) and the filtering means (3). These communication means (5) allow the cartridge (1) to be fluidly connected to a device in which it is to be incorporated (e.g. a device according to the invention as described below). The fluid communication means (5) is adjacent to the filtering means. The fluid communication means (5) and the filtering means (3) allow the liquid sample to pass through the chamber from upstream to downstream.

[0099] The chamber (4) consists of a solid stationary phase of the reversed-phase C18 type for retaining (for example by adsorption) the radiolabeled compound. The solid stationary phase thus advantageously allows a preliminary purification of the sample. Filtering means (3) (by the casing) keep the solid stationary phase within the volume of the chamber, whilst filtering out potential solid particles to allow the passage of the liquid sample (thus advantageously preventing clogging of the column during subsequent purification by high pressure chromatography).

[0100] FIG. 2 shows a device (6) for the pre-purification and injection of a liquid sample containing a radiolabeled compound in a high pressure chromatography system (7). This device (6) comprises a 6-way multiway valve (8). This multiway valve (8) allows for the fluidic connection (12) of all the elements of the device, in particular the supply means (9), the elution phase pump (10), the cartridge (1) and the discharge means (11). The valve also allows for the fluidic connection (12) of the high pressure chromatography system (7) to the device of the invention. The supply means (9) allows for the supply of the device (6) with a liquid sample of a radiolabeled compound, for example from a radiosynthesis module. The elution phase pump (10) is used to aspirate the elution phase in the device (6). The cartridge (1) allows for the pre-purification of the liquid sample containing the radiolabeled compound.

[0101] 3a and 3b illustrate the operation of the device (6) of FIG. 2 and an embodiment of the method of the present invention.

[0102] Figure 3a shows the loading step. In this first step, the multi-way valve (6-port valve) (8) is in the loading position (13). The loading position (13) allows the supply means (9) to supply a liquid sample containing the radiolabelled compound to the cartridge (1). As the sample passes through the cartridge (1), the radiolabelled compound is retained on the solid stationary phase. This advantageously allows the radiolabelled compound to be concentrated and / or pre-purified at this stage of the process. Components (14) that are not retained by the cartridge (1) (i.e. those that have little affinity for the stationary phase) flow past the cartridge through the multi-way valve (8) to the discharge means (11).

[0103] The loading position can also be used to carry out a second loading step by feeding the cartridge (1) via the feeding means (9) with a second liquid fraction comprising radiolabeled compounds (e.g. a fraction resulting from rinsing / washing with a washing phase for the radiosynthesis module located upstream of the feeding means (9)). In this case, the second liquid fraction thus further "loads" the cartridge already loaded with radiolabeled compounds from the liquid sample fed in the first loading step. In this second loading phase (which can be repeated), radiolabeled compounds, for example originating from washing the synthesis module, are recovered in the cartridge, helping to improve the radiosynthesis yield. Meanwhile, the washing phase and other elements not retained in the cartridge are removed in the discharge means (11). This allows washing of the radiosynthesis module and the feeding means in order to recover as much radiolabeled compounds as possible without affecting the final amount of sample injected into the chromatography system (7). In fact, this amount can be selected / controlled in the following injection step.

[0104] Furthermore, in this loading step, the elution phase (15) can be sent directly to the chromatography system (7) via the multi-way valve (8) in the usual way, which allows balancing of the chromatography column during loading of the cartridge (1).

[0105] FIG. 3b shows the injection step. In the second step of the process, the multi-way valve (8) is in the injection position (16). The injection position (16) allows the radiolabeled compounds retained in the cartridge (1) to be eluted and the enriched and / or pre-purified sample to be injected into the chromatography system (7) for purification (or analysis). This also allows the sample to be pre-filtered through the cartridge (1), which prevents clogging of the chromatography column. In this injection step, the pump (10) draws the elution phase (15) through the multi-way valve (8) towards the cartridge (1), which releases and elutes the radiolabeled compounds retained in the cartridge (1). The elution phase consisting of the radiolabeled compounds (17) is then injected into the high pressure chromatography system (7) through the multi-way valve (8). The elution phase consisting of the radiolabeled compounds (17) thus constitutes an enriched and / or pre-purified sample compared to the initial sample (before passing through the cartridge). In particular, the initial volume can be adapted (eg reduced), the solvent can be changed, certain elements (elements not retained by the cartridge) can be removed, and solid particles present can be filtered out.

[0106] Furthermore, during this injection step, the supply means (9) are in fluid communication with the discharge means (11), which allows the supply means (9) to be cleaned and rinsed before the next loading step, which has the advantage of reducing the risk of subsequent contamination and / or the formation of agglomerates that may block the system and require maintenance.

[0107] It is to be understood that the invention is in no way limited to the embodiments described above, but modifications can be made without departing from the scope of the claims. It is further understood that the invention also encompasses all possible combinations of the features and preferred features described herein and claimed.

[0108] Additionally, the following examples are provided for illustrative purposes and are not intended to limit the scope of the present invention.

[0109] This example illustrates the use of the compound [ 18 F]-AV-45 by automated purification using the cartridges, devices and methods of the present invention.

[0110] Made of PEEK, volume 785mm 2 The cartridge according to the invention was manually packed with 90 mg of stationary phase of HLB type (hydrophilic-lipophilic balance, Waters HLB Plus Short cartridge reference number 186000132) with a particle size of 50-65 microns.

[0111] Automated radiosynthesis (capture of fluorine-18 onto a QMA cartridge, elution of fluorine-18, drying of fluorine-18 in the reactor, labeling, hydrolysis, neutralization) was performed in the first module (NEPTIS®×Seed™) with 2.0 mg of precursor in 2.0 mL of DMSO.

[0112] The neutralized reaction stock solution was manually diluted to 10 mL with water in a 10 mL Luer lock syringe. This was manually placed into a syringe pump which automatically loaded the cartridge according to the invention. The syringe pump speed was 4 mm / 6 sec and a relative vacuum of -0.9 bar was applied. Once loading was complete, without rinsing the syringe, the operator activated the injection valve (8) in injection mode (according to FIG. 2) via the NEPTIS® software.

[0113] The UV trace was recorded using the SFC module software with simultaneous manual acquisition initiation.

[0114] To monitor the fluorine-18 labeled product, a radio detector was attached to the cartridge and a second detector was placed at the outlet of the SFC purification system, which allowed the operator to activate a collection valve to collect the radioactive peak in a collection vessel.

[0115] In this example, the radioactivity contained in the 10 mL syringe before loading into the cartridge was 11.4 mCi (420 Mbq).

[0116] The "semi-prep" column used was a Phenomenex Polar RP 10 μm, 250×10 mm column.

[0117] The SFC conditions were as follows: -Elution: CO2 / EtOH95 / 5; -Flow rate: 20mL / min; -150 bar, pressure regulator heated to 60°C; -UV detection at 254nm; -SFC system temperature: 25~30℃

[0118] Measurements showed that 90% of the activity on the cartridge was eluted in 1 minute with an eluent of CO2 / EtOH 95 / 5.

[0119] Compound [ 18 The radioactive peak associated with [F]-AV-45 appears between 6'20'' and 7'30''. For comparison, the HPLC purification showed 18 F]-AV-45 usually elutes near the 15′ position.

[0120] At the end of harvest, the last container was manually removed from the harvest module and inserted into the activity meter to measure its activity.

[0121] The results of this example are shown in the table below (measurements of residual radioactivity and synthesis yield during the process (AO: initial radioactivity, dc: "decay-corrected value" corrected for the decay of fluorine-18).

[0122]

Table 1

Claims

1. A cartridge (1) for the pre-purification of a liquid sample containing a radiolabeled compound intended to be purified or analyzed by high-pressure chromatography, said cartridge (1) comprising: a casing (2); a filtering means (3); a chamber (4) whose volume is limited by said casing (2) and said filtering means (3) and which contains a solid stationary phase; and fluid communication means (5) upstream and downstream of said chamber (3) and said filtering means (3), The cartridge (1), wherein said casing (2) and said fluid communication means (5) are mechanically capable of withstanding fluid pressures of 50 bar or more.

2. 2. A cartridge (1) according to claim 1, characterized in that the casing (2) and the communication means (5) are mechanically capable of withstanding fluid pressures of 100 bar or more.

3. 3. Cartridge according to claim 1 or 2, characterized in that the casing (2) and / or the communication means (5) are made of a material selected from PEEK or stainless steel.

4. 2. The cartridge of claim 1, wherein the solid stationary phase is of the reversed phase type.

5. 2. The cartridge according to claim 1, wherein the volume of the chamber (4) is between 50 and 2000 mm ３ A cartridge characterized by:

6. 1. An apparatus for pre-purifying and injecting (6) a liquid sample containing a radiolabeled compound into a high-pressure chromatography system (7), said apparatus comprising: a multi-way valve (8); a supply means (9) for a liquid sample containing a radiolabeled compound; an elution phase pump (10); A cartridge (1) according to claim 1, and a discharge means (11), The valve is in fluid communication (12) with the supply means (9), the pump (10), the cartridge (1), the discharge means (11) and the high-pressure chromatography system (7).

7. 7. Device according to claim 6, characterized in that the chromatography system (7) is of the HPLC or SFC type.

8. 8. Device according to claim 6 or 7, characterized in that the chromatography system (7) is of the SFC type.

9. 7. A method for pre-purifying and injecting a liquid sample containing a radiolabeled compound into a high-pressure chromatography system (7) using the device according to claim 6.

10. 1. A method for pre-purifying and injecting a liquid sample containing a radiolabeled compound into a high-pressure chromatography system (7), said method comprising the steps of: (i) a loading step; (ii) an injection step, The loading step comprises supplying a liquid sample containing a radiolabeled compound to the cartridge (1) according to claim 1 via a multi-way valve (8) at a loading position (13); retaining said radiolabeled compound by said cartridge (1); and discharging elements (14) not held by the cartridge (1) via the multi-way valve (8) to a discharging means (11), The injecting step aspirating the elution phase (15) towards the cartridge (1) through the multi-way valve (8) in the injection position (16) by means of a pump (10); Releasing the radiolabeled compound held in the cartridge (1); and injecting the elution phase containing the radiolabeled compound (17) through the multi-way valve (8) into a high-pressure chromatography system (7).

11. 10. The method according to claim 9, characterized in that the high pressure chromatography system (7) is of the HPLC or SFC type.

12. 10. The method according to claim 9, characterized in that the high pressure chromatography system (7) is of the SFC type.

13. 10. The method according to claim 9, characterized in that the elution phase (15) is ethanol.

14. 11. A method according to claim 10, comprising, after the step of injecting, a step of regenerating the cartridge (1), said steps being, in the following order: supplying a regeneration phase through a multi-way valve (8) in the loading position (13) of the cartridge (1); Discharging said phases through said multi-way valve (8) to a discharge means (11).

15. 10. The method of claim 9, wherein the regeneration phase is selected from water, an aqueous buffer, and an organic solvent. A method characterized by:

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