Molecular sensors, methods and kits for detecting and quantifying carbon dioxide - Patents.com

Abstract

The present invention relates to a sensing method for specifically detecting, measuring and / or monitoring carbon dioxide. It further relates to a probe configured to specifically sense carbon dioxide. The probe is of formula (I'), where Dy is a dye moiety, Preac (Preact) is a moiety reactive to carbon dioxide, and L is a spacer. The present invention also relates to a kit specifically configured for sensing, detecting and / or measuring carbon dioxide.

Description

[Technical field]

[0001] Technical area The present invention relates to a means for determining the presence of carbon dioxide (CO2) in a test sample. It further relates to reagents that enable the specific detection of carbon dioxide, as well as test kits specifically configured for detecting, measuring and monitoring the presence of carbon dioxide in a sample. [Background technology]

[0002] Related Technology Carbon dioxide is currently detected by a wide variety of methods, including non-dispersive infrared (NDIR) sensors, positron emission tomography (PET) imaging, Severinghaus electrodes, paper-based sensors, hydrogels, polymers, and MOFs. Most of these methods are not well suited for measuring carbon dioxide at the scale of laboratory experiments with high selectivity and sensitivity, thus hindering rapid, accurate, and reliable analysis of biological or chemical samples and reactions.

[0003] Activity-based sensing (ABS) has been used to sense low molecular weight reactive molecules, making it possible to measure their contribution to several biological and chemical reactions. According to this principle, the analyzed low molecular weight molecule is involved in a chemical reaction such that it can be identified by a unique spectral measurement. Reactive nitrogen and oxygen species are successfully identified and monitored using ABS. Reactive carbonyl species such as carbon monoxide, phosgene and formaldehyde are also detected by this means. However, a suitably specific, sensitive and accurate sensor for carbon dioxide has not been developed so far.

[0004] Thus, there is scope for the development of new and improved sensors specifically configured to identify and measure carbon dioxide. Summary of the Invention

[0005] BRIEF DISCLOSURE OF THE PRESENT ART It is therefore an object of the present invention to provide a method suitable for specifically sensing, monitoring and / or detecting the presence of carbon dioxide in a sample. In particular, the method is for selectively detecting, sensing or measuring carbon dioxide in the presence of other reactive carbon species (RCS), such as carbon monoxide, carbon disulfide and formaldehyde. The object is to achieve a selectivity of carbon dioxide detection over other reactive carbon species of more than 10, preferably more than 100, more preferably more than 1000. Furthermore, the object is to achieve a sensitivity of carbon dioxide detection of a few ppm, such as 10 ppm.

[0006] Another object of the present invention is to provide functional molecules that are highly reactive towards carbon dioxide as well as highly specific, while at the same time allowing its easy detection by direct means such as visual or spectral parameters.

[0007] Another object of the present invention is to provide a tool and / or kit or test arrangement specially adapted for measuring the presence of carbon dioxide in a sample, in particular during a biological or chemical process or reaction. It is a further object to accurately monitor the production of carbon dioxide and to measure the concentration variations over time in such a sample. The object is to provide such a kit or test arrangement which requires a limited number of analytical tools and steps.

[0008] Another object of the method is to provide a ready-to-use diagnostic method that allows to identify and / or characterize the amount of carbon dioxide, or its occurrence, in a biological sample.

[0009] According to the invention, these objects are achieved by means of the objects of the independent claims (further detailed in the dependent claims).

[0010] In accordance with what is known in the art, the present invention provides precise means and products specifically configured to sense, measure and monitor the presence of carbon dioxide. [Brief description of the drawings]

[0011] Exemplary embodiments of the invention are disclosed in the specification and illustrated by the following drawings: [Figure 1] FIG. 2 is a graph of absorbance and fluorescence of a compound according to the invention with and without carbon dioxide. [Diagram 2] FIG. 2 is a graph showing the correlation between signal intensity and carbon dioxide concentration for the compounds according to the present invention. [Diagram 3] FIG. 1 is a diagram of the comparative reactivity of compounds according to the invention towards carbon dioxide and other reactive carbon species. [Figure 4a] FIG. 2 shows the effect of reactive moieties in the compounds according to the present invention. [Figure 4b] FIG. 2 shows the effect of reactive moieties in the compounds according to the present invention. [Figure 4c] FIG. 2 shows the effect of reactive moieties in the compounds according to the present invention. [Figure 5a] FIG. 1 shows the effect of the dye portion of the compound according to the present invention. [Figure 5b] FIG. 1 shows the effect of the dye portion of the compound according to the present invention. [Figure 6] An example of detecting the inhibitory effect of an enzyme reaction using the compound according to the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Examples of embodiments of the present invention The present disclosure provides a method or process specifically adapted for the detection of carbon dioxide in a sample. A sample is understood herein as any part or subpart of a material under test. The sample can be under various physical conditions such as gas, liquid, gel, suspension, emulsion or related conditions. It can be, for example, an air or gas sample, a liquid solution or a liquid suspension. The sample can be an aliquot or part of an environment such as seawater or air, or a part of a reaction mixture, in which a certain amount of carbon dioxide is sensed and measured. In addition, a sample can also refer to any evolving environment, such as a reaction mixture, in which the variation of carbon dioxide is sensed and monitored over time, especially as a continuous detection. A sample can also encompass a production means in which the creation of carbon dioxide needs to be detected and / or monitored, either continuously or for a scheduled period of time.

[0013] The sensing process involves a chemical reaction of carbon dioxide with a reagent used as a sensing molecule. For this reason, it is considered an activity-based sensing (ABS) method or process. In particular, the reaction rate between carbon dioxide and such a reagent should be as high as possible, so that very small amounts of carbon dioxide and / or small amounts of reagent can be used. Furthermore, it is preferred that the reaction is carried out at ambient temperature, so that no heating or activation step is required. Thereby, the reaction preferably occurs at temperatures of 20°C, 10°C, 5°C or lower. In a preferred embodiment, the reaction occurs efficiently throughout a wide temperature range, such as from -10°C to 80°C or from about 0°C to about 50°C. According to another preferred embodiment, the sensitivity to carbon dioxide does not change significantly over the above temperature range. In other words, the sensitivity changes by no more than 5% or no more than 10%. Additionally or alternatively, the specificity or selectivity to other reactive carbon species does not change significantly throughout the previously mentioned temperature range. In particular, it changes by no more than 10% or no more than 5%.

[0014] According to an embodiment, the sensing process or method requires only one reagent to be used as the sensing molecule, meaning that the reagent itself reacts spontaneously in situ with carbon dioxide to provide a ready-to-read result. Preferably, no co-reactant or catalyst is required. Thus, the sensing method does not require the addition of other chemicals.

[0015] The reagent, preferably used as a single reagent, may be commercially available and supplied as such, so that it can be used as is for carbon dioxide measurement, detection or monitoring operations. Thus, the reagent is sufficiently stable at room temperature or at a controllable temperature comprised between 5° C. and 30° C., or 10° C. and 30° C. Under such circumstances, the method does not include any preparation steps allocated to prepare the functional reagent.

[0016] Alternatively, the reagent may be provided as two or more different separate subparts. This may be required for stability reasons, for safety reasons, or for other reasons such as regulatory constraints. In particular, each separate subpart of the reagent may correspond to a dye moiety or dye moiety precursor, a reactive moiety or reactive moiety precursor, or a linker or a linking precursor configured to adsorb the reagent to a surface, or any other suitable subpart of the reagent. Preferably, all these subparts are combined together to create a functional reagent. Preferably, the combination of the subparts is easily handled and can be achieved simply by mixing the subparts in a common solvent or by solubilizing one subpart in the other. Under these conditions, the sensing method may include a preliminary step of preparing a functional reagent based on the predetermined subparts of the reagent. According to a specific embodiment, several subparts are provided as a kit, where different dye moieties can be selected to react with different reactive moieties, so that a diversity of reagents is generated upon carbon dioxide measurement. For example, depending on the specific needs, the user may select one dye moiety or one dye precursor through the selection of several dye moieties or dye precursors and combine it with a predetermined reactive moiety or reactive precursor. Furthermore, the reactive moiety or reactive precursor may be selected through the selection of several possible reactive moieties or reactive precursors. Thus, the method may include a step of selecting one or both of the dye and reactive moiety of the reagent, followed by a step of combining them and generating the corresponding functional reagent. If other subparts are available, they may be equally subject to a preliminary selection through several options. Thus, the subparts associated with the spacer that connects two different subparts of the reagent are selected through several options. The linker configured to bind the reagent to the surface may be selected through several options, for example, depending on the nature of the support. This allows many variations that are not specifically described herein to be available.Thus, it will be appreciated that the sensing method or process may be capable of adapting or adjusting the reagents upon measurement or detection.

[0017] The step of combining the subparts may, if appropriate, include the addition of one or more additives that allow for easy reaction between the subparts. Such additives may relate to catalysts, buffers or other suitable adjuvants. Preferably, the preparation of the reagent based on the subparts is carried out at ambient temperature, such as comprised between 5° C. and 30° C. or 10° C. and 25° C., by simple manual stirring in a vial. It preferably occurs instantly or within a time frame of a few minutes, such as less than 5 minutes or less than 15 minutes.

[0018] Preparation of the reagent may be carried out prior to the carbon dioxide measurement or detection operation, and advantageously it can be done on-site, in which case the sensing method or process includes adding a sub-part of the reagent to or in contact with the sample, instead of the reagent itself, so that carbon dioxide detection or measurement can commence as soon as the reagent is produced.

[0019] The reagent used as the sensing molecule is selective or specific for carbon dioxide. This means that the reaction rate is clearly statistically significantly different compared to other reactive carbon species such as carbon monoxide, carbon disulfide, and formaldehyde. The reaction rate may be, for example, at least 10, 100, 1000, or 10,000 times higher for carbon dioxide compared to other reactive carbon species. In a preferred embodiment, since it is specific for carbon dioxide, it does not cause the desired fluorescence with other reactive carbon species. This makes the sensing method or process applicable to samples that contain several reactive carbon species, in addition to the carbon dioxide sensed, in particular one or more of carbon monoxide, formaldehyde, and carbon disulfide. Furthermore, the sensing method is also applicable to reaction mixtures with reactive species other than carbon dioxide, without disturbing the reaction itself. Thus, the method allows to follow up a reaction, such as a chemical, enzymatic, or biological reaction, without disturbing it.

[0020] The reagent of the present invention comprises a reactive moiety configured to react spontaneously with carbon dioxide. According to a preferred embodiment, the reactive moiety is designed to react irreversibly with carbon dioxide so as to ensure its capture under reactive conditions. By reactive conditions is meant herein the conditions under which the detection or measurement of carbon dioxide is carried out. This offers the advantage of limiting or avoiding any exchange or equilibrium with other reactive carbon species potentially present in the sample. The sensing method or process comprises a step of contacting such a reagent or one of its subparts with a sample such that the reagent reacts with the carbon dioxide present in the sample.

[0021] According to a preferred embodiment, the reactive moiety of the reagent is a phosphorus-based reactive moiety Preac. Preferably, the reactive moiety is a phosphine, such as a triarylphosphine, such as triphenylphosphine, an alkyldiarylphosphine, such as methyldiphenylphosphine or ethyldiphenylphosphine, or a dialkylarylphosphine, such as dimethylphenylphosphine, and related chemical reagents. By alkyl is meant herein any linear or branched alkyl containing 1 to 10 carbon atoms. Thus, they include the groups methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, cyclohexyl, adamantyl, etc. The phosphorus-based reactive moiety Preac equally means a phosphine that does not contain an aromatic group and carries only alkyl groups, such as those mentioned above. In the phosphorus-based reactive moiety Preac, each one of the mentioned groups, either the alkyl or the aromatic, may be independently substituted by one or more groups, such as halogen, NO2, CF3, NH2, SO2, COH, COOH, or related groups.

[0022] More preferably, the phosphorus-based reactive moiety Preac is attached to the amine of the reagent to form an iminophosphorane. Thus, the phosphorus-based reactive moiety Preac can be easily displaced by the group CO by reaction with carbon dioxide. The reactivity of the phosphorus-based reactive moiety Preac towards carbon dioxide, including its reaction rate and its selectivity with respect to other reactive carbon species, can be adjusted or tuned by the substituents of the phosphorus-based reactive moiety Preac. For example, one or more of the phenyl groups of the phosphines mentioned above can independently contain one, two, three or four substituents such as nitro, halogen, CF3, carbonyl, sulfoxide or related chemical groups.

[0023] Upon reaction of carbon dioxide with the reagent, an intramolecular reaction occurs, leading to the substitution of the phosphorus-based reactive moiety Preac by the group CO. The intramolecular reaction is designed to provide a visual change in the reagent molecule, as described in more detail below. Preferably, the intramolecular reaction is carried out under reaction conditions between the CO group originating from the ambient carbon dioxide and a reactive group or heteroatom present in the reagent molecule. Such heteroatoms or reactive groups can be selected from among amines, such as secondary amines, alcohols, thiols, or any suitable reactive chemical group. The sensing process or method thus comprises modifying the reagent in situ by reaction with carbon dioxide to reveal its presence in the sample. The intramolecular reaction can mean any suitable intramolecular reaction configured to structurally modify the core structure of the reagent. It can also mean, for example, a ring closure or rearrangement, preferably a ring closure. Although intermolecular reactions are not completely excluded, they are currently considered to be unfavourable.

[0024] The reagent used in the sensing process or method further comprises a dye moiety. A dye moiety Dy is understood herein as a part of a reagent that exhibits optical and / or spectral activity. Any optically related activity may be considered with respect to the properties of the dye moiety Dy. Such optical and / or spectral properties may relate, for example, to light absorption, emission, phosphorescence, and fluorescence in a particular spectral range, a change in a particular wavelength, or a color in the visible, IR, or UV range. Other optically related properties such as polarization shift or Raman effect may also be considered. Although optical and / or spectral parameters are preferred, the method may be based on other measurable parameters as well, such as magnetic related parameters. It is understood herein that it is preferable that the properties of the dye moiety Dy can be measured directly from the sample. This thus eliminates any separate analytical steps with chemical separation, chemical purification, and associated analytical additional steps.

[0025] The properties of the dye Dy of the reagent are designed to change upon reaction of the reagent with the surrounding carbon dioxide. More particularly, the properties of the dye moiety Dy change upon the intramolecular reaction mentioned above, leading to a change in the core molecular structure and affecting the properties of the dye moiety. According to a preferred embodiment, the dye moiety Dy is directly bonded to the reactive group or heteroatom involved in the intramolecular reaction. The term "direct bond" means herein that the optical and / or spectral properties of the dye moiety Dy remain localized as close as possible to the reactive group or heteroatom involved in the intramolecular reaction so as to be modified by the structural change resulting from the intramolecular reaction. This excludes, for example, any spacer between the dye moiety Dy and the reactive group or heteroatom involved in the intramolecular reaction.

[0026] In a preferred embodiment, the reactive group or heteroatom involved in the intramolecular reaction is an amine, and the dye moiety Dy contains at least one double bond in the alpha position relative to the amine to which it is attached. Under such a configuration, the double bond is attached to the free lone pair of electrons of the nitrogen atom of the amine. The dye moiety Dy preferably contains other unsaturations, such as delocalized double or triple bonds, which are responsible for the optical and / or spectral properties, etc. In particular, the dye moiety Dy preferably contains one or more aromatic rings, such as phenyl, naphthyl, pyridine, quinoline, quinoxaline, phenanthrene, and related aromatic elements.

[0027] The sensing process or method thus comprises the step of contacting a reagent as described above, where the dye moiety Dy has a first optical and / or spectral property for a sample containing carbon dioxide, where the reagent specifically or selectively reacts with carbon dioxide and further engages in an intramolecular reaction resulting in a modified reagent, where the dye moiety Dy has a second optical and / or spectral property. The first optical and / or spectral property and the second optical and / or spectral property are different from each other such that the modified reagent can be easily and immediately sensed.

[0028] According to a preferred embodiment, the optical and / or spectral characteristic is related to fluorescence. The first optical and / or spectral characteristic may consist of non-fluorescence or fluorescence at a first wavelength. The second optical and / or spectral characteristic may consist of the presence of fluorescence or stronger fluorescence or fluorescence at a second wavelength different from the first wavelength. Preferably, the dye moiety Dy of the reagent does not show fluorescence at a predefined wavelength, such as 455 nm or another specific wavelength, and shows measurable fluorescence at this specific wavelength when the reagent reacts with carbon dioxide. According to such an arrangement, the presence of carbon dioxide in the sample switches ON the fluorescence of the reagent, which can be detected. Strictly speaking, one or more wavelengths can be considered as determining "switch ON" when fluorescence appears at said specific wavelength.

[0029] The reverse arrangement can also be envisaged. For example, the first optical and / or spectral characteristic can consist of fluorescence at a predetermined wavelength, which is switched off upon reaction with carbon dioxide. In this configuration, when all the reagents have reacted, the fluorescence is not visible. This makes it possible, for example, to detect that a predetermined amount of carbon dioxide has been consumed based on a predetermined amount of reagent.

[0030] Thus, the sensing process or method comprises detecting one or both of the first and second optical properties of the dye moiety Dy. For this purpose, a suitable spectral or optical measuring device may be used. If only the presence of carbon dioxide is intended to be detected in the sample, no specific optical device is required when carbon dioxide switches the reagent ON. A simple visual control is sufficient to determine the presence of carbon dioxide. Also, a color shift may occur, which is visually identified. Such a method is conveniently configured to easily detect the appearance of carbon dioxide, either for safety reasons or for monitoring the course of a chemical process. When the first and second optical and / or spectral parameters consist of two different wavelengths or two different spectral profiles, then a detection device may be required.

[0031] According to another embodiment, the first and / or second optical and / or spectral properties are correlated to the amount of carbon dioxide in the sample. For example, the intensity of the corresponding optical or spectral signal is proportional to or related to the concentration of carbon dioxide. Thus, the sensing process or method includes measuring the amount of carbon dioxide or continuously monitoring its occurrence in the sample. Continuous monitoring means that continuous measurements are performed without interruption. Alternatively, the monitoring is performed stepwise by sensing and measuring carbon dioxide at scheduled time periods and comparing the values ​​obtained over time.

[0032] Thus, the step of detecting either or both of the first and second optical properties of the dye moiety Dy means either simply detecting the presence of carbon dioxide in the sample or detecting its presence and measuring its concentration over time. Carbon dioxide is detected when it reaches a certain amount or a certain concentration. The method allows for the detection of very small amounts of carbon dioxide percentages, below 10 ppm or as low as a few ppm, such as 5 ppm or 1 ppm.

[0033] The detection step is carried out using any suitable detection device, such as a confocal microscope or a plate reader.

[0034] The sensing method or process according to the invention may use two or more of the reagents described herein. All of the steps described above are equally applicable to one reagent or two different reagents used simultaneously. For example, a first reagent may be selected for its improved sensitivity to carbon dioxide, thereby allowing detection of low concentrations of carbon dioxide. If monitoring must be performed over a wide range of carbon dioxide concentrations, a second reagent that is more accurate for higher carbon dioxide concentrations may be used in combination. Similarly, detecting or measuring carbon dioxide over a wide range of pressures and / or temperatures may require the use of different reagents that show a specific response in a particular subrange with respect to temperature or pressure. This can result in a more accurate detection and / or measurement of carbon dioxide. In that case, the two different reagents may be selected based on various parameters such as their stability, sensitivity, selectivity, solubility, etc.

[0035] According to another embodiment, the above described method or process may be repeated sequentially using different reagents.

[0036] Depending on the nature of the experiment, the sensing process may include one or more additional steps, such as: - Dissolving a reagent in a solvent to prepare a solution with a predetermined concentration of the reagent. The reagent may be supplied as a powder or as a liquid. As a result, it may not be possible to use it to detect or monitor carbon dioxide. Solubilizing it in a suitable solvent makes it possible to prepare a calibration solution whose optical and / or spectral properties can be directly sensed and monitored. When the concentration of the reagent is known, the amount of carbon dioxide sensed can be determined. The solvent may be water, distilled water, a dedicated solvent such as dimethyl sulfoxide, ethanol, or other dedicated solvents and their mixtures. - Adsorption of reagents onto a solid surface such as a glass or paper surface. For this purpose, a reagent solution can be sprayed onto the solid support or the surface can be immersed in the reagent solution. Other means such as capillary action can be used to bring the reagents from the solid support to react with the sample.

[0037] According to a preferred embodiment, the process for the detection and / or monitoring of carbon dioxide in a sample comprises treating said sample with a reagent of formula (I'): [ka] When reacted with ambient carbon dioxide, a compound of formula (II'): [ka] contacting the mixture to obtain During the ceremony: -Dy means a dye moiety having first optical and / or spectral properties in a compound of formula (I) and second optical and / or spectral properties in a compound of formula (II); -Preac refers to a phosphorus-based reactive moiety, where CO2 reacts in an irreversible manner, -L denotes an internal spacer selected from a linear or branched carbon chain, which is either saturated or mono-, di- or tri-unsaturated, containing 1 to 10, preferably 1 to 6, carbon atoms, or a cyclic chain, which is either saturated or mono- or di-unsaturated, containing 4 to 7 or 5 to 6 carbon atoms, or an aromatic group such as phenyl, or a heteroaromatic group such as pyridine, (benzo)furan, (benzo)thiophene or pyrrole, where in the flexible linear or branched carbon chain and in the non-aromatic cyclic chain, where applicable, 1 to 3 carbon atoms may be optionally replaced, independently of one another, by a heteroatom selected from -O-, -S-, -NH-, -CO-, -NHCO- or -COO-, or by the group -OH, -NH2, halogen, NO2 or alkyl containing 1 to 3 carbon atoms, -R1 represents 1 to 4 groups independently selected from H, C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, or represents 2 groups which together with the spacer L to which they are linked form a fused saturated, unsaturated or aromatic C4-C8 ring, in which 1 to 3 carbon atoms, if applicable, are optionally replaced, independently of one another, by groups selected from -COO-, -CO-, NH-, -O-, -S-, or by groups selected from -OH, SH, NH2, NO2, halogen, or represents a macromolecule such as DNA, RNA, peptides, proteins, enzymes, and -n means an integer selected from 0, 1, 2 or 3.

[0038] According to a more preferred embodiment, the process for detecting and / or monitoring carbon dioxide in a sample comprises treating said sample with a compound of formula (I') {wherein the spacer L is a phenyl group. That is, a compound of formula (I): [ka] When the reagent of formula (II) reacts with ambient carbon dioxide, a compound of formula (II): [ka] contacting the mixture to obtain In the formula, Dy, Preac, R1, and n are as defined above. Preferably, in formulas (I) and (II), R1 represents 1 to 4 groups independently selected from H, C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, or represents two groups which together with the phenyl group to which they are linked form a fused saturated, unsaturated or aromatic C4-C8 ring. Preferably, in formulae (I) and (II), n means 0, 1 or 2, more preferably 0 or 1.

[0039] According to a specific embodiment, R1 in any compound of formula (I) and (I') can mean or include a linker configured to attach the reagent to a solid surface, such as a glass, metal or cellulose surface. Such a linker can include a group at its free end that is reactive to -SiOH, -SH, -OH, or related groups present on the solid surface. For this purpose, the linker can carry a group at its free end, such as a carbonyl, an aldehyde, an amine, or any suitable reactive group such that a covalent bond can be established.

[0040] Alternatively, one of the R1 groups in the compounds of formula (I) and (I') represents a macromolecule such as DNA, RNA, a peptide, a protein, an enzyme or any biochemical macromolecule.

[0041] Alternatively, one or more of the R groups may be configured to improve the solubility of the reagent in some specialized solvents. Thus, the C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, and the previously mentioned fused saturated, unsaturated or aromatic C4-C8 rings may be substituted with polar groups such as -OH, SH, NH2, NO2, halogens, and related groups to improve solubility in water or polar solvents.

[0042] The dye moiety Dy contains at least one double bond alpha to the nitrogen atom to which it is attached, such that said double bond is bonded to the free lone pair of electrons of the nitrogen atom in the compound of formula (I), (I').

[0043] The dye moiety Dy may be any of the following moieties Dy1, Dy2 and Dy3: [ka] is preferably selected from one of the following:

[0044] The group Preac means any phosphorus-based reactive moiety mentioned above, in particular triarylphosphine, alkyldiarylphosphine, dialkylarylphosphine or trialkylphosphine, where alkyl and aryl are as defined herein. According to a preferred embodiment, for the reactive moiety, the group Preac means a phosphine selected from PMePh2, PEtPh2, PMe2Ph and PPh3, where "Ph" independently means an unsubstituted phenyl group or a phenyl group substituted with 1 to 4 substituents selected from linear or branched C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, NO2, halogen and CF3.

[0045] In the reagent, the reactive moiety Preac and the reactive group or heteroatom involved in the intramolecular reaction are linked through a spacer L configured to allow the intramolecular reaction. Such a spacer may be a flexible linear or branched carbon chain containing 1 to 10 carbon atoms, as disclosed in formulae (I), (II) and (III), or a cyclic chain of 2 to 6 carbon atoms, preferably containing 4 to 6 carbon atoms, more preferably containing an aromatic group such as a phenyl or pyridine group. In the flexible linear or branched carbon chain and in the non-aromatic cyclic chain, where applicable, 1 to 3 carbon atoms may be replaced, independently of each other, optionally by a heteroatom selected from -O-, -S-, -NH-, -CO-, -NHCO- or -COO-, or by the group -OH, -NH2, halogen, NO2 or alkyl containing 1 to 3 carbon atoms.

[0046] When reacted with ambient carbon dioxide, the compound of formula (I) forms an intermediate compound of formula (III): [ka] wherein Dy, R1, and n are as defined above, which immediately provides an intramolecular reaction leading to a compound of formula (II). The intramolecular reaction contributes to an optical and / or spectral change of the dye moiety Dy. Preferably, both the reaction with carbon dioxide and the intramolecular reaction are irreversible.

[0047] Of course, the reaction is equally applicable to linkers L other than phenyl groups, so long as intramolecular reaction is possible. Thus, compounds of formula (I') can be prepared by reacting with intermediates of formula (III'): [ka] wherein L, Dy, R1, and n are as defined above.

[0048] The present invention also includes a reagent as described above, which is specialized to react specifically with carbon dioxide and provide a visible structural change in situ upon reaction with carbon dioxide. To this end, it comprises a reactive moiety and a dye moiety whose properties are modified upon reaction with carbon dioxide. It further comprises a reactive group or heteroatom configured to provide an intramolecular reaction upon reaction of said reactive moiety with carbon dioxide. The reactive group or heteroatom is linked to the reactive moiety through a suitable spacer L. The reactive moiety is preferably a phosphorus-based reactive moiety Preac (Preact) as defined above. The dye moiety is preferably one of the dye moieties Dy as defined above.

[0049] As mentioned above, any combination of dye moiety Dy and reactive moiety Preac can be envisaged to tailor the reagent according to needs.

[0050] In a preferred embodiment, the reagent of the present invention is the following (I1), (I2), (I3), (I4), (I5) and (I6): [ka] is selected from among:

[0051] The reagents can be prepared by reacting a precursor of the dye moiety with a precursor of the reactive moiety, linked together by a suitable spacer L, and a building block containing a reactive group or a heteroatom.

[0052] For example, for a reagent related to a compound of formula (I), a compound of formula (Ia): [ka] is a compound of formula (Ib): [ka] to produce a compound of formula (Ic): [ka] wherein Dy, R1 and n are as defined above and wherein X represents a suitable electrophile such as a halogen, preferably a Cl or Br atom, a carbonyl, a thiocarbonyl, a triflate, a mesylate or a group O-SO2-R, wherein R represents any suitable substituent such as linear, branched or cyclic alkyl, which contains 1 to 10 carbon atoms and is optionally substituted, or an aryl group.

[0053] Such reactions may be carried out in a suitable solvent such as ethanol, DMSO, DMF, ethyl acetate, water, etc. Suitable additives such as bases may be used. For example, the above reaction may be carried out in the presence of triethylamine in ethanol at temperatures between 60°C and 120°C for 10 minutes to 20 hours.

[0054] The resulting compound contains a precursor of a reactive moiety, which needs to be transformed into a suitable reactive moiety by one or more steps. For the reagent related to the compound of formula (I), the process comprises reacting the compound of formula (Ic) with the phosphorus-based moiety Preac, as defined above, in DMSO at a temperature between 20° C. and 80° C. to obtain the corresponding compound of formula (I) mentioned above.

[0055] The present invention further relates to a kit specially adapted for the detection, measurement and / or monitoring of carbon dioxide. The kit according to the invention comprises at least one reagent as described above. Preferably, the kit comprises two or more reagents, each adapted to different test conditions or having a different response. For example, a reagent may be considered unsuitable if its optical or spectral properties overlap with those of the sample itself such that the optical or spectral change is not easily detectable. The kit may comprise at least some instructions regarding the optical and / or spectral properties of the reagent(s). It may additionally comprise information related to the correlation of the optical or spectral signal with the carbon dioxide concentration. The kit may comprise, for one or more reagents, corresponding reagent subparts supplied separately to enable their combination.

[0056] The reagent or its subparts may be provided using any suitable form, such as a liquid in a vial, or a solid, powder or gel in a suitable receiving solid. The reagent or its subparts may be provided in a predetermined amount corresponding to a particular use. For example, it may be provided as a compressed solid having a predetermined weight and easily dispersible in the sample. Alternatively or additionally, the kit may include one or more solvents or may include one or more vials configured to receive solvents, each having a predetermined size, so that the reagent may be prepared in a predetermined concentration. Alternatively, the reagent or some of the reagents of the kit may be pre-adsorbed on a solid surface, such as a glass or paper-based surface. Thus, contacting the solid surface with the sample may provide a visual change of the surface, such as the appearance of a color, and thus indicate the presence of carbon dioxide.

[0057] The kit may additionally include a light emitting device, such as a UV light emitting device, configured to illuminate the sample and reveal the responsive signal of the reagent. EXAMPLES

[0058] Example 1: Spectral measurements Compound (I1) was dissolved in DMSO and subjected to a carbon dioxide detection experiment, and the compound of formula (I1b): [ka] (I1) and (I1b) exhibited different absorbance and emission maxima, making them suitable for CO2 detection conditions. Figure 1 shows the signals observed for both (I1) and (I1b) at 455 nm in the presence of high concentrations of carbon dioxide. Compound (I1) was placed in the presence or absence of CO2 (Figure 2). In the absence of carbon dioxide, negligible fluorescence response was observed, whereas upon bubbling of carbon dioxide, a strong fluorescence response that further enhanced over time was detected with an emission maximum at 455 nm, matching the spectrum of (I1b). The turn-on response saturated at a signal-to-noise ratio of 260. This high value demonstrates the high sensitivity of the fluorescence approach. Example 2: Kinetics Experiments The reaction kinetics was investigated with the same compounds of formulae (I1) and (I1b) by measuring the fluorescence at different time points using different concentrations of either (I1) or carbon dioxide. The reaction showed a first-order dependence on both (I1) and carbon dioxide, pointing towards the initial intermolecular isocyanate formation as the rate-limiting step. A gas mixture with various ratios of CO2 / N2 was bubbled into a solution containing (I1) at a constant volume and rate (controlled by a mass flow controller). (I1) showed a CO2 dose-dependent response. It could sense a mixture of 1.5% CO2 with a signal-to-noise of ~20. Both polar and non-polar solvents, including water, were used without any dispersion in the results, making it sufficiently specific for a wide range of interdisciplinary applications. The compound of formula (I2) was used to sense atmospheric CO2 levels. (I2) was exposed to an air / N2 mixture. The detection limit (background + 3σ) was 1.5% air (98.5% N2) mixture. Since air contains ~0.04% CO2, this indicates that (I2) can detect carbon dioxide in the gas mixture at a concentration of 0.0006% (~6.4 ppm). Example 3: Selectivity Experiments The selectivity of (I1) was compared to a panel of relevant reactive carbon species shown in Figure 3. Compound (I1) did not produce any significant fluorescence response to most of the reactive carbon species tested other than CO2, thus demonstrating selectivity for CO2 over other more reactive compounds. The reactive, potentially competing small molecules mentioned in Figure 3 were: [Table 1] After 60 min, CO2 was added to all samples and luminescence was measured again. The successful activation of the sensor by CO2 in the presence of the tested reactive carbon species indicates that, apart from trisphosgene, which led to a complex mixture of products, these compounds did not adversely affect and interfere with the sensor. Example 4: Effect of the reactive moiety Preac The ability to tune the reactivity of the sensor to carbon dioxide was investigated. The compound of formula (I1) mentioned above was mixed with the compounds of formulae (I6) and (I3) below: [ka] Figures 4a, 4b and 4c provide comparative data regarding their reactivity towards carbon dioxide. In these figures, the corresponding compounds (I2b) and (I3b) correspond to compound (I1b), in which the PPh3 group is replaced by a PPhMe2 and a PPh2Me group, respectively. Example 5: Effect of dye moiety Dy Two molecules of formula (I2) and (I4) below: [ka] was added to a carbon dioxide sensing experiment, and the results are shown in Figures 5a and 5b. Example 6: Detection of Carbon Dioxide in Liquid Samples The compound of formula (I1) was added to a microscale experiment of catalytic decarboxylation reaction. The benzylic decarboxylation process was investigated using a standard protocol in which the solvent contained 100 μM (I1). During the course of the reaction, the released carbon dioxide was captured by the sensor, generating a fluorescent signal. Fluorescence measurements showed an almost instantaneous reaction. In the presence of catalytic amounts of base, para-nitrophenylacetic acid was converted to the corresponding decarboxylation product. In the absence of base, no product was obtained. Reactions carried out in the presence of base resulted in decarboxylation and showed a significant increase in the fluorescent signal compared to those in the absence of base. In addition, time-dependent experiments demonstrated a correlation between the generated fluorescent signal and the reaction progress, as monitored by HPLC. Example 7: Enzymatic monitoring Fluorescence real-time monitoring of enzyme activity was investigated for screening of inhibitors and drugs. Urease, a nickel-based metalloenzyme that converts urea to ammonia and CO2, was used in the presence of the compound of formula (I2) mentioned above (water-soluble and brightly fluorescent in water). The enzyme assay was carried out in PBS at pH=7.4 with varying concentrations of both urea and urease (from Canavalia ensiformis (Jack bean)). 96-well plates were used for the setup and the fluorescence response was measured over time using a plate reader. FIG. 6 shows that the sensor was able to distinguish all of the altered levels of both urea and urease, while only negligible signal was observed in the absence of either urea or urease. Example 8: Cell Imaging The selectivity of compound (I4) was evaluated against common analytes of cells. HeLa cells were treated with compound (I4) [5 μM] for 10 min, followed by washing three times to remove excess sensor. The cells were then incubated under ambient air or 5% CO2 as an exogenous CO2 model, and imaged at various time points. A significant time-dependent increase in fluorescence was observed in the red channel and a decrease in the green channel. Measurement of the fluorescence signal occurring within the cells was found to exactly match the spectral fingerprint of the activated sensor, confirming the reaction of compound (I4) with CO2. In addition, the time-dependent response of cells incubated under ambient conditions was attributed to basal levels of CO2 production by metabolic processes. Experimental details

[0059] All reagents used were purchased from commercial suppliers and used as generally accepted unless otherwise noted. Solvents used in synthesis were of puriss grade and technical grade solvents were used for column chromatography.

[0060] All reactions were carried out in oven- or flame-dried glassware equipped with Teflon-coated magnetic stir bars unless otherwise noted. Reactions were monitored by thin layer chromatography (TLC) unless otherwise noted. TLC was performed on Merck silica gel 60 F254 TLC aluminum plates and visualized using UV fluorescence quenching and / or potassium permanganate (KMnO4) staining.

[0061] Column chromatographic purification was performed as flash column chromatography (FCC) using silica gel (SiliCycle, 40-63 μm or Sigma Aldrich 40-60 μm) and an applied air pressure of 0.3-0.5 bar. The yields obtained refer to the chromatographically purified compounds unless otherwise indicated.

[0062] NMR spectra were recorded at room temperature on a Bruker Avance III 300 MHz, a Bruker Avance III HD 400 MHz, a Bruker Neo 400 MHz or a Bruker Neo 500 MHz spectrometer. 1 H and 13 The C spectrum was used as a reference for residual solvent peaks. Multiplicities are abbreviated as s (singlet), d (doublet), t (triplet), q (quartet), p (quintet), hepto (septet), m (multiplet) and combinations thereof. High resolution mass spectrometry (HRMS) data were obtained using electrospray ionization (ESI) on a Bruker maXis-ESI-Qq-TOF-MS and are reported in m / z units. Coumarin-based probes: [ka] AMC-N3 [ka] AMC-Cl (200.0 mg, 795 μmol, 1.0 equiv) and (2-azidophenyl)methanamine 2,3 (147 mg, 993 μmol, 1.25 equiv) were suspended in EtOH (2 ml) in an 8 ml vial, and NEt3 (0.554 ml, 3.97 mmol, 5.0 equiv) was added. The mixture was heated to 100° C. for 3 h and monitored by TLC (EtOAc:hex 30:70). Upon complete conversion of the starting material, the resulting precipitate was filtered, washed with EtOH, and dried under high vacuum to give AMC-N3 (286 mg, 787 μmol) as a white solid in 99% yield. 1 H NMR (400 MHz, DMSO-d6) δ 7.91 (t, J = 6.0 Hz, 1H), 7.83 (d, J = 9.2 Hz, 1H), 7.45 - 7.30 (m, 2H), 7.30 - 7.22 (m, 1H), 7.22 - 7.11 (m, 1H), 6.66 (dd, J = 9.1, 2.6 Hz, 1H), 6.39 (d, J = 2.6 Hz, 1H), 4.68 (s, 1H), 4.36 (d, J = 5.7 Hz, 2H), 3.41 (q, J = 6.9 Hz, 4H), 1.11 (t, J = 6.9 Hz, 6H). 13 C NMR (101 MHz, DMSO-d6) δ 162.17, 155.40, 153.93, 150.18, 137.12, 128.73, 128.53, 128.12, 125.04, 123.43, 118.73, 107.79, 102.09, 96.92, 78.72, 43.84, 41.12, 12.33. AMC-NH2 [ka] AMC-Cl (50 mg, 0.2 mmol, 1.0 equiv) and 2-(aminomethyl)aniline (97 mg, 0.8 mmol, 4.0 equiv) were suspended in EtOH (2 ml) in an 8 ml vial and NEt3 (111 μL, 0.8 mmol, 4.0 equiv) was added. The mixture was heated to 100° C. for 3 h and monitored by TLC (EtOAc:hex 50:50). After completion, the reaction mixture was diluted with EtOAc (100 ml) and washed with 0.1 M HCl (50 ml) and brine (50 ml). The organic layer was separated, dried over Na2SO4, and evaporated under reduced pressure. The crude product was purified by column chromatography (EtOAc:hex 50:50) to give AMC-NH2 (31 mg, 0.1 mmol) as a white solid in 46% yield. 1 H NMR (400 MHz, chloroform-d) δ 7.19 (q, J = 9.2, 8.3 Hz, 3H), 6.80 (td, J = 7.4, 1.2 Hz, 1H), 6.78 - 6.75 (m, 1H), 6.49 (s, 2H), 5.29 (s, 1H), 5.01 (t, J = 4.7 Hz, 1H), 4.31 (d, J = 4.5 Hz, 2H), 3.95 (s, 2H), 3.38 (q, J = 7.1 Hz, 4H), 1.18 (t, J = 7.1 Hz, 6H). 13 C NMR (101 MHz, CDCl3) δ 164.43, 162.91, 155.92, 154.14, 150.78, 145.26, 130.65, 129.92, 121.31, 120.44, 119.11, 116.54, 108.10, 102.30, 98.36, 81.36, 77.16, 45.14, 44.80, 12.57. AMC-PPh3 [ka] AMC-N3 (50 mg, 0.14 mmol, 1.0 equiv) and triphenylphosphine (40 mg, 0.15 mmol, 1.1 equiv) were suspended in DMSO (2 mL) in an 8 mL vial. The mixture was heated to 50° C. and monitored by TLC (EtOAc:hex 50:50). After completion, the reaction mixture was diluted with EtOAc (50 ml) and washed with brine (25 mL). The organic layer was separated, dried over Na2SO4, and evaporated under reduced pressure. The crude product was purified by column chromatography (EtOAc:hex 50:50) to give AMC-PPh3 (72 mg, 0.12 mmol) as an off-white solid in 87% yield. 1 H NMR (400 MHz, chloroform-d) δ 7.74 - 7.66 (m, 6H), 7.56 - 7.49 (m, 3H), 7.46 - 7.39 (m, 6H), 7.20 (ddd, J = 7.4, 2.7, 1.7 Hz, 1H), 6.95 (s, 1H), 6.87 (td, J = 7.6, 1.8 Hz, 1H), 6.71 - 6.62 (m, 2H), 6.47 (dt, J = 8.0, 1.2 Hz, 1H), 6.43 (d, J = 2.6 Hz, 1H), 6.04 (dd, J = 9.0, 2.6 Hz, 1H), 5.31 (d, J = 7.1 Hz, 1H), 4.56 (d, J = 4.5 Hz, 2H), 3.31 (q, J = 7.1 Hz, 4H), 1.13 (t, J = 7.1 Hz, 6H). 13 C NMR (101 MHz, CDCl3) δ 164.71, 155.91, 154.14, 150.14, 149.81, 132.49, 132.10, 131.33, 130.54, 130.32, 129.94, 129.03, 128.46, 121.52, 121.27, 117.64, 107.50, 103.37, 98.28, 79.86, 46.88, 44.68, 12.58. 31P NMR (162 MHz, CDCl3) δ 4.68. AMC-PPh2Me [ka] AMC-N3 (50 mg, 0.14 mmol, 1.0 equiv) and methyldiphenylphosphine (28 μL, 0.15 mmol, 1.1 equiv) were suspended in DMSO (2 ml) in an 8 mL vial. The mixture was heated to 50° C. and monitored by TLC (EtOAc). After completion, the reaction mixture was diluted with EtOAc (50 ml) and washed with brine (25 mL). The organic layer was separated, dried over Na2SO4, and evaporated under reduced pressure. The crude product was purified by column chromatography (EtOAc) to give AMC-PPh2Me (31 mg, 0.06 mmol) as an off-white solid in 43% yield. 1 H NMR (400 MHz, DMSO-d6) δ 8.00 (t, J = 5.9 Hz, 1H), 7.91 (ddt, J = 12.0, 6.8, 1.5 Hz, 4H), 7.76 (d, J = 9.1 Hz, 1H), 7.63 - 7.47 (m, 6H), 7.05 (dt, J = 7.6, 2.2 Hz, 1H), 6.77 (td, J = 7.6, 1.8 Hz, 1H), 6.61 (dd, J = 9.1, 2.6 Hz, 1H), 6.49 (td, J = 7.3, 1.1 Hz, 1H), 6.38 (d, J = 2.5 Hz, 1H), 6.32 (dt, J = 7.9, 1.2 Hz, 1H), 5.00 (s, 1H), 4.62 (d, J = 5.8 Hz, 2H), 3.40 (q, J = 7.0 Hz, 4H), 2.32 (d, J = 12.7 Hz, 3H), 1.11 (t, J = 7.0 Hz, 6H). 13C NMR (101 MHz, DMSO-d6) δ 162.44, 155.42, 154.07, 149.99, 149.47, 132.90, 131.90, 131.63, 130.99, 130.89, 130.44, 130.22, 128.88, 128.76, 127.14, 127.05, 123.20, 119.84, 119.72, 116.13, 107.64, 102.46, 96.93, 78.60, 59.75, 43.82, 42.70, 39.52, 20.76, 14.08, 13.18, 12.56, 12.35. 31 P NMR (162 MHz, DMSO-d6) δ 3.93. AMC-PPhMe2 [ka] AMC-N3 (7.8 mg, 1.000 Eq, 21 μmol) and dimethylphenylphosphine (4.3 mg, 4.0 μL, 1.00 Eq, 21 μmol) were suspended in DMSO (1 ml) in an 8 ml vial and shaken at room temperature to obtain a homogeneous solution. The resulting product, AMC-PPhMe2, was used as is without further purification. 11H NMR (400 MHz, DMSO-d6) δ 8.01 (t, J = 6.0 Hz, 1H), 7.88 (ddt, J = 11.6, 6.7, 1.5 Hz, 2H), 7.75 (d, J = 9.1 Hz, 1H), 7.62 - 7.50 (m, 3H), 7.03 (dt, J = 7.7, 2.1 Hz, 1H), 6.81 (td, J = 7.6, 1.8 Hz, 1H), 6.64 (dd, J = 9.1, 2.6 Hz, 1H), 6.48 (td, J = 7.3, 1.1 Hz, 1H), 6.38 (tt, J = 3.2, 1.3 Hz, 2H), 5.03 (s, 1H), 4.48 (d, J = 5.9 Hz, 2H), 3.40 (q, J = 7.0 Hz, 4H), 1.94 (d, J = 13.0 Hz, 6H), 1.11 (t, J = 6.9 Hz, 6H). 13 13C NMR (101 MHz, DMSO-d6) δ 162.48, 155.41, 153.96, 150.17, 149.98, 133.53, 132.65, 131.45, 131.42, 130.41, 130.31, 130.15, 129.98, 129.75, 128.79, 128.68, 128.31, 128.26, 127.78, 127.31, 127.08, 123.13, 119.54, 119.41, 115.71, 107.66, 102.50, 96.96, 78.63, 43.81, 42.64, 39.52, 15.54, 14.84, 13.95, 13.82, 12.34. 31 31P NMR (162 MHz, DMSO-d6) δ 8.49. AMC-Dye

Chem.

Chem.

Claims

1. CO in the sample 2 1. A method for the detection of providing a reagent comprising a moiety reactive towards carbon dioxide and a dye moiety Dy, wherein the dye moiety Dy has first optical and / or spectral properties; - contacting the sample with the reagent so that the reagent reacts with carbon dioxide to provide a reacted agent, wherein the dye moiety Dy has second optical and / or spectral properties that differ from the first optical and / or spectral properties; and - detecting one or both of the first and second optical and / or spectral properties of the dye moiety Dy; Including, wherein the reagent is a compound of formula (I'): 【Chemistry 1】 and the reacted reagent is a compound of formula (II'): 【Chemistry 2】 And, During the ceremony: -Dy means a dye moiety having a first optical property in a compound of formula (I') and a second optical property in a compound of formula (II'); - Preac (Preact) means a phosphorus-based reactive moiety, and where CO 2 reacts in an irreversible manner, -R1 is H, C 1 -C 6 Alkyl, C 1 -C 6 Alkenyl, C 1 -C 6 alkynyl, or together with the spacer L to which they are connected, a fused saturated, unsaturated or aromatic C 4 -C 8 means two groups forming a ring, in which 1 to 3 carbon atoms are optionally substituted, if applicable, independently of one another, by a group selected from -COO-, -CO-, NH-, -O-, -S-, or by -OH, SH, NH 2 , NO 2 , substituted with a group selected from halogen), or macromolecules such as DNA, RNA, peptides, proteins, enzymes, etc. -L denotes an internal spacer selected from a linear or branched carbon chain, which is either saturated or mono-, di- or tri-unsaturated, containing 1 to 10, preferably 1 to 6, carbon atoms, or a cyclic chain, which is either saturated or mono- or di-unsaturated, containing 4 to 7 or 5 to 6 carbon atoms, or an aromatic group such as phenyl, or a heteroaromatic group such as pyridine, (benzo)furan, (benzo)thiophene, or pyrrole, where, where applicable, in the flexible linear or branched carbon chain and in the non-aromatic cyclic chain, 1 to 3 carbon atoms may optionally be replaced, independently of one another, by a heteroatom selected from the group -O-, -S-, -NH-, -CO-, -NHCO- or -COO-, or 2 , halogen, NO 2 or alkyl containing 1 to 3 carbon atoms, and - n means an integer selected from 0, 1, 2 or 3, The method.

2. 2. The method of claim 1, wherein the dye moiety Dy comprises at least one double bond alpha to the nitrogen atom to which it is attached, such that the double bond is bonded to the free lone electron pair of the nitrogen atom in formula (I').

3. The dye moiety Dy may be any of the following moieties Dy1, Dy2 and Dy3: 【Transformation 3】 2. The method of claim 1, wherein the ion exchange reaction is selected from one of the following:

4. The dye moiety Dy is selected from the group consisting of the following moieties: Dy1, Dy2 and Dy3: 【Chemistry 4】 3. The method of claim 2, wherein the ion exchange reaction is selected from one of the following:

5. The group Preac (Preact) is PMePh 2 , PMe 2 Ph and PPh 3 where "Ph" is an unsubstituted phenyl group or C 1 -C 6 Alkyl, C 2 -C 6 Alkenyl, C 2 -C 6 Alkynyl, NO 2 , halogens and CF 3 The method of claim 1, wherein each independently means a phenyl group substituted with 1 to 4 substituents selected from:

6. The method of claim 2, wherein the group Preac (Preact) means a phosphine selected from PMePh 2 , PMe 2 Ph and PPh 3 , where "Ph" independently means an unsubstituted phenyl group or a phenyl group substituted with 1 to 4 substituents selected from C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, NO 2 , halogen and CF 3 .

7. The method of claim 3, wherein the group Preac (Preact) means a phosphine selected from PMePh 2 , PMe 2 Ph and PPh 3 , where "Ph" independently means an unsubstituted phenyl group or a phenyl group substituted with 1 to 4 substituents selected from C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, NO 2 , halogen and CF 3 .

8. The method of claim 4, wherein the group Preac (Preact) means a phosphine selected from PMePh 2 , PMe 2 Ph and PPh 3 , where "Ph" independently means an unsubstituted phenyl group or a phenyl group substituted with 1 to 4 substituents selected from C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, NO 2 , halogen and CF 3 .

9. The aforementioned CO 2 with a compound of formula (I) to produce an intermediate compound of formula (III'): 【Transformation 5】 wherein Dy, L, R1, and n are as defined in any one of claims 1 to 5, and which immediately provides an intramolecular reaction leading to a compound of formula (II').

10. 2. The method of claim 1, wherein the first and second optical properties comprise one or more of fluorescence, luminescence, and light absorption at specific wavelengths, and the wavelengths differ between the first and second optical properties.

11. At least one of the first and second optical properties is 2 The method of claim 1, wherein the amount of

12. The method of claim 1 , wherein the sample is selected from a liquid sample, an air sample, or a gas sample.

13. 10. The method of claim 1, wherein the sample is a biological, biochemical, or chemical sample, and the method allows for sensing variations in carbon dioxide over time.

14. Compounds of formula (I'): 【Transformation 6】 And, During the ceremony: -Dy means a dye moiety having a first optical property in a compound of formula (I) and a second optical property in a compound of formula (II); - Preac (Preact) means a phosphorus-based reactive moiety, and where CO 2 reacts in an irreversible manner, -R1 is H, C 1 -C 6 Alkyl, C 1 -C 6 Alkenyl, C 1 -C 6 alkynyl, or together with the phenyl group to which they are attached, a fused saturated, unsaturated or aromatic C 4 -C 8 means two groups forming a ring, and -L denotes an internal spacer selected from a linear, branched or cyclic carbon chain containing 1 to 10, preferably 1 to 6 carbon atoms, which is either saturated or mono-, di- or tri-unsaturated, or an aromatic group selected from phenyl, or a pyridine group; and - n means an integer selected from 0, 1, 2 or 3, The compound.

15. The compound is selected from the group consisting of: 【Transformation 6】 15. The compound of claim 14, selected from:

16. A method for preparing a compound of formula (I') according to any one of claims 14 or 15, comprising the steps of: In the presence of triethylamine in ethanol at a temperature of 60° C. to 120° C. for 10 minutes to 20 hours, a compound of formula (Ia′): 【Transformation 8】 with a compound of formula (Ib): 【Chemistry 9】 to produce a compound of formula (Ic'): 【Chemistry 10】 To provide Including, wherein Dy, R1, L and n are as defined in claim 14, and wherein X represents a suitable electrophile such as halogen, carbonyl, thiocarbonyl, triflate, mesylate and related electrophiles. The method.

17. 17. The method of claim 16, further comprising reacting the compound of formula (Ic') of claim 15 with a phosphorus-based moiety Preac in DMSO at a temperature between 20°C and 80°C to obtain the compound of formula (I') above, wherein Dy, R1, L and Preac are as defined in claim 14.

18. 16. A kit for testing for the presence and / or amount of carbon dioxide in a sample, said kit comprising at least one compound of formula (I') according to claim 14 or 15.

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