A group for the detection of analytes

Abstract

A group (1) for the detection of analytes comprising a containment chamber (2), a separation device (3) housed inside the containment chamber (2) and a collection screen (4) also housed inside the containment chamber (2). The separation device (3) comprises (a) a plurality of circular filters (5) arranged together in a coaxial position along a vertically arranged X axis in use and (b) a cylindrical side wall (6), within which the plurality of circular filters (5) is housed and in which at least one window (8) with controlled opening is obtained and arranged at one or more of the circular filters (5). Each of the circular filters (5) has a filtering ability lower than that of the circular filter below and is adapted to rotate around the X axis. The collection screen (4) comprises a collection surface (9) facing the plurality of windows.

Description

[0001] "A GROUP FOR THE DETECTION OF ANALYTES"

[0002] Cross-Reference to Related Applications

[0003] This Patent Appl ication claims priority from Italian Patent Applications No . 102023000022761 filed on October 30 , 2023 , the entire disclosure of which is incorporated herein by reference .

[0004] Technical Field

[0005] The present invention relates to a group for the detection of analytes , the use o f which is advantageously but not solely applied to the analysis of metabolites and active ingredients in biological fluids for clinical and pre-clinical studies of new drugs .

[0006] State of the Art

[0007] Clinical and pre-clinical studies of new drugs and medical devices provide for the quantitative analysis of active ingredients and metabolites thereof present in the biological fluid following administration .

[0008] The main techniques used hitherto relate to the chromatographic analysis with mass detector and a technique referred to as LBA, an English acronym which stands for Ligand Binding Assay .

[0009] For these techniques , there is the guideline ICHM10 , which defines the criteria which must be met in order for the obtained data to be taken into account and thus be signi ficant .

[0010] Both the above techniques are limited to only researching and quanti fying the substances under study, for which the standards thereof must necessarily be available .

[0011] In particular, the LBS technique provides for the use of enzymes or reagents capable o f selectively binding the molecule of interest and make it visible with chemical and physical procedures , such as , for example , fluorescence and gelation . The need for using a selective bind necessarily implies that a speci fic reagent is sought and available , as well as the creation of a calibration curve .

[0012] The chromatographic technique (HPLC, GC or UPLC ) with mass detector is a more precise technique , but it suf fers from the issue related to the selectivity of the analyte sought with respect to multiple variables due to matrix ef fects and proper detection technique .

[0013] The variability of the biological matrix, as well as the variability of the sample , requires that it is subj ected to a series of treatments before analysis , with apparent disadvantages in terms of productivity and loss of part of the sample that this entails .

[0014] In addition, the analysis techniques used hitherto must be generally adapted depending on the fluid of analysis , living species being analyzed, and also the presence of diseases ( e . g . , glucose or cholesterol ) which could interfere with the analysis .

[0015] Finally, it should be considered that since the analysis techniques of the prior art analyze a single analyte at a time , i f more than one analyte from the same sample is to be studied, one is forced to have to repeat the analysis technique for each of them .

[0016] Description of the Invention

[0017] A need was thus felt for an analysis technique whose technical features could overcome the drawbacks of the prior art .

[0018] An obj ect of the present invention is a group for the detection of analytes comprising a containment chamber, a separation device housed inside the containment chamber and a collection screen also housed inside the containment chamber ; said separation device comprising ( a ) a plurality of circular filters arranged together in a coaxial position along a vertical ly arranged X axis in use ; each of the circular filters being structured to retain larger particles than those retained by the circular filter immediately below and being able to rotate around the X axis ; and (b ) a cylindrical side wall , within which the said plurality of circular filters is housed and in which at least one window with controlled opening and arranged at one or more of said circular filters ; said collection screen comprising a collection surface facing said plurality of windows .

[0019] Preferably, the collection surface has a curvature parallel to that of said cylindrical side wall .

[0020] Preferably, in the cylindrical side wall a plurality of windows with controlled opening are obtained, each of which is arranged at a respective circular filter .

[0021] Preferably, in the side wall a single window with controlled opening is obtained and extending along all the circular filters .

[0022] Preferably, the collection screen is made of a material adapted to allow a spectroscopic analysis of the collected analytes .

[0023] Preferably, the separation device comprises means adapted to creating an axial and unidirectional flow of an inert gas through the said plurality of circular filters .

[0024] Preferably, each of the said circular filters comprises a concave peripheral portion adapted to house the molecules pushed by the centri fugal force towards the outside of the filter itsel f . Preferably, in the containment chamber there is an inert atmosphere at a pressure equal to or lower than that present inside said cylindrical side wall .

[0025] Preferably, the distance between the collection screen and the cylindrical side wall is adj ustable .

[0026] Brief Description of the Drawings

[0027] For a better understanding of the present invention, here below is described an implementation example for illustrative and non-limiting purposes together with the accompanying figures , in which :

[0028] - Figure 1 is an extremely schematic perspective view with parts removed for clarity of a part of the group according to the present invention;

[0029] - Figure 2 is an extremely schematic top view with parts removed for clarity of the group according to the present invention and in which operating principles are depicted; and

[0030] - Figure 3 is an extremely schematic perspective view with parts removed for clarity of the embodiment and in which operating principles are depicted .

[0031] Preferred Embodiment of the Invention

[0032] In the accompanying figures , numeral 1 indicates , as a whole , a detection group according to the present invention .

[0033] The group 1 comprises a ( schematically illustrated) containment chamber 2 , a separation device 3 housed inside the containment chamber 2 and a collection screen 4 also housed inside the containment chamber 2 .

[0034] The separation device 3 compri ses a plurality of circular filters 5 arranged one on top of the other equally spaced apart in a coaxial position along a vertically arranged X axis in use and a cylindrical side wall 6 arranged in order to contain the plurality of circular filters 5 .

[0035] The circular filters 5 exhibit an increasing filtering ability from top to bottom . This means that a circular filter 5 retains particles larger than those retained by the next circular filter 5 . Preferably, a steady flow of inert gas is created inside the cylindrical side wall 6 which pushes the biological sample downwards . The creation of such a flow is achieved by inert gas insuf flation means arranged at an upper cylindrical portion 6 , and suction means arranged at a lower cylindrical portion 6 .

[0036] Each of the circular filters 5 comprises a concave peripheral portion 7 , which has the function of housing the molecules ( analytes ) subj ected to the centri fugal force as it will be described below .

[0037] In fact , each of the circular filters 5 is arranged to rotate about the X axis and, consequently, subj ects the molecules it retained to a centri fugal force which moves them towards its concave peripheral portion 7 . In this manner, the molecules move until they come in contact with the concave peripheral portion 7 by arranging themselves inside it .

[0038] In the cylindrical side wall 6 , a longitudinal window 8 is obtained extending at all the circular filters 5 . The window 8 is equipped with controlled opening .

[0039] As an alternative to what described above , a plurality of windows can be obtained in the cylindrical side wall 6 , each of which is arranged at a single circular filter 5 . In this manner it wi ll be possible to discriminate the opening of one or more windows while keeping the others closed .

[0040] As illustrated in Figure 2 , taking a single circular filter 5 into consideration, when all the molecules will be arranged in the concave peripheral portion 7 and will all have the same angular velocity previously set, the sudden opening of the window 8, together with the blocking of the rotation of the circular filters 5, causes the molecules to leak through the window 8. Each molecule leaks out of the window 8 with a trajectory which will result from the angular velocity (or) and centrifugal force (mrw2) indicated with two respective arrows in Figure 2.

[0041] The tangential velocity will be the same for all molecules and depends on the angular velocity of the circular filter 5; while the centrifugal force depends on the molecule mass and is a function of the square of the angular velocity, which increases the difference between the masses of the molecules. Since the radius of the circular filters cannot be changed during the analysis, the instrumental variable changing the selectivity of the instrument is the angular velocity .

[0042] Based on what reported above, it is evident that the molecules retained by a single circular filter 5 (of the same size) leak out of the relative window of the cylindrical side wall with different trajectories depending on their mass. As depicted in Figure 2, at the same tangential velocity, the molecules with higher mass (schematically illustrated with a solid circle) will have a trajectory which is more affected by the centrifugal force compared to a molecule with lower mass (schematically illustrated with an empty circle) . The molecules of the same size (retained by the same circular filter 5) are placed on top of a collection surface 9 of the collection screen 4 in different positions depending solely on mass.

[0043] As will be readily apparent, the distance between the cylindrical side wall 6 and the collection screen 4 is the second variable affecting the instrument selectivity.

[0044] Another aspect relevant for the molecules passing from the separator device 3 to the detection screen 4 refers to the pressure and atmosphere found in the containment chamber 2. Specifically, the containment chamber 2 must have an inert atmosphere and must be at an equal to or lower pressure than that present inside the separation device 3. As is readily apparent to a person skilled in the art, the inert atmosphere can be created by using an inert gas or by vacuum.

[0045] As illustrated in Figure 3, repeating the same pattern above for all circular filters 5 results in a selection of molecules by size and mass.

[0046] If the surface 9 of the collection screen 4 is plotted in a two-dimensional graph, a diagonal distribution of molecules is obtained. Molecules having equal size but different mass (differing in atoms bound together but occupying the same dimensional space) will be placed on the same Y ordinate, as they are retained by the same circular filter, but on a different X axis, as they have different atomic masses.

[0047] Likewise, it occurs that molecules of equal masses may be retained by different filters depending on the position they take when they come in contact with the filter. In this case, the molecules will be placed on the same X axis (mass) but on different Y axis (size) . Such an evidence allows to further characterize the molecule.

[0048] Based on what described above, it is evident that the analysis performed by the group of the present invention does not suffer from any matrix effects and, consequently, it does not require any prior treatments. Furthermore, the molecule analysis does not provide for having any internal standards, and molecules with different chemical and physical characteristics can be analyzed simultaneously .

[0049] A further characterization occurs if the collection screen 4 is made of a clear, UV-visible diode array detector (DAD) material. In this manner, the entire UV spectrum of each particle collected by the collection screen 4 is detected. In this manner, with a good database, each molecule would be easily identified by mass, size and UV spectrum. The use of UV, thanks to Beer-Lambert law, also enables to quantify the entire composition of the biological sample.

[0050] The absorption measurement of the molecules having equal mass and UV spectrum could also measure the amount of molecules present in the sample without any interference.

[0051] For substances which are not visible in the UV-visible spectrum, such as some inorganic salts, one could change the type of collection screen 4 by allowing the use of other techniques such as fluorescence, RAMAN, NIR, or IR.

[0052] It may also be assumed to place a layer on the collection screen 4 which promotes selective growth for biological tests.

[0053] Since the analysis performed by the group of the present invention is non-destructive, the treated molecules may also be subsequently subjected to other chemical / physical (e.g., NMR, IR) or biological analysis techniques.

[0054] In summary, the detection group of the present invention, in addition to being unaffected by the matrix effect, is capable of simultaneously analyzing the (polarizable and non-polarizable ) substances present in a sample of biological liquid in a short time, without any loss of materials . In this manner, potential correlations between substances and metabolites that were not selectively searched for before can be detected . The kinetics of side reactions to the drug can also be detected on metabolites not previously searched for, thus being able to anticipate potential side ef fects of new drugs by reducing the number of patients involved .

[0055] Further advantages from the use of the detection group according to the present invention refer to the possibility of searching for the analytes on di f ferent body fluids ( e . g . , urine and plasma ) without changing the procedure ; and the possibility of searching for the analytes on samples of di f ferent living species and comparing the data obtained; they can be compared with each other since the procedure is the same .

[0056] Finally, with the detection group of the present invention, the times for sample development and analysis are greatly reduced compared to the prior art , and the creation of a database with the data acquired each time would allow an immediate analysis of all analytes on emergency patients , extending the use of such instruments from clinical research to routine clinical analysis .

Claims

C L A I M S1. A group (1) for the detection of analytes comprising a containment chamber (2) , a separation device (3) housed inside the containment chamber (2) and a collection screen (4) also housed inside the containment chamber (2) ; said separation device (3) comprising (a) a plurality of circular filters (5) arranged together in a coaxial position along a vertically arranged X axis in use; each of the circular filters (5) being structured to retain particles larger than those retained by the circular filter (5) immediately below and adapted to rotate around the X axis; and (b) a cylindrical side wall (6) , within which the said plurality of circular filters (5) is housed and in which at least one window (8) with controlled opening and arranged at one or more of said circular filters (5) ; said collection screen (4) comprising a collection surface (9) facing said plurality of windows.

2. The group for the detection of analytes according to claim 1, characterized in that said collection surface has a curvature parallel to that of said cylindrical side wall (6) .

3. The group for the detection of analytes according to claim 1 or 2, characterized in that in said cylindrical side wall (6) a plurality of windows with controlled opening are obtained, each of which is arranged at a respective circular filter ( 5 ) .

4. The group for the detection of analytes according to claim 1 or 2, characterized in that in said side wall a single window (8) with controlled opening is obtained and extending along all the circular filters (5) .

5. The group for the detection of analytes according toone of the previous claims, characterized in that said collection screen (4) is made of a material adapted to allow a spectroscopic analysis of the collected analytes.

6. The group for the detection of analytes according to one of the previous claims, characterized in that said separation device (3) comprises means adapted to create an axial and unidirectional flow of an inert gas through the said plurality of circular filters (5) .

7. The group for the detection of analytes according to one of the previous claims, characterized in that each of the said circular filters (5) comprises a concave peripheral portion (7) adapted to house the molecules pushed by the centrifugal force towards the outside of the filter (5) .

8. The group for the detection of analytes according to one of the previous claims, characterized in that in said containment chamber (2) there is an inert atmosphere at a pressure equal to or lower than that present inside said cylindrical side wall (6) .

9. The group for the detection of analytes according to one of the previous claims, characterized in that the distance between said collection screen (4) and said cylindrical side wall (6) is adjustable.

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