CA1142841A - Process and a reagent for the determination of ions and of polar and/or lipophilic substances in fluids - Google Patents
Process and a reagent for the determination of ions and of polar and/or lipophilic substances in fluidsInfo
- Publication number
- CA1142841A CA1142841A CA000336828A CA336828A CA1142841A CA 1142841 A CA1142841 A CA 1142841A CA 000336828 A CA000336828 A CA 000336828A CA 336828 A CA336828 A CA 336828A CA 1142841 A CA1142841 A CA 1142841A
- Authority
- CA
- Canada
- Prior art keywords
- process according
- entity
- complex
- chromophore
- host molecule
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C10—PETROLEUM, GAS OR COKE INDUSTRIES; TECHNICAL GASES CONTAINING CARBON MONOXIDE; FUELS; LUBRICANTS; PEAT
- C10M—LUBRICATING COMPOSITIONS; USE OF CHEMICAL SUBSTANCES EITHER ALONE OR AS LUBRICATING INGREDIENTS IN A LUBRICATING COMPOSITION
- C10M161/00—Lubricating compositions characterised by the additive being a mixture of a macromolecular compound and a non-macromolecular compound, each of these compounds being essential
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/84—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving inorganic compounds or pH
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/04—Endocrine or metabolic disorders
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/805—Optical property
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/807—Apparatus included in process claim, e.g. physical support structures
- Y10S436/808—Automated or kit
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S436/00—Chemistry: analytical and immunological testing
- Y10S436/815—Test for named compound or class of compounds
Abstract
ABSTRACT OF THE DISCLOSURE
The present invention provides a process for the determination of ions or of polar and/or lipophilic substances in liquids, wherein the ion or the polar and/or lipophilic substance to be determined is allowed to act upon a complex ligand or host molecule which is selective towards the ion or the polar and/or lipophilic substance to be determined, said complex liquid or host molecule being attached by a direct covalent bond or heteropolar bond or hydrogen bridge or hydrophobic bond to a chormophore or containing a chromo-phore in the form of an inclusion complex, whereafter the change of extinction or the wavelength displacement is measured; the present invention also provides a reagent for carrying out this process, as well as a test kit containing the reagent.
The present invention provides a process for the determination of ions or of polar and/or lipophilic substances in liquids, wherein the ion or the polar and/or lipophilic substance to be determined is allowed to act upon a complex ligand or host molecule which is selective towards the ion or the polar and/or lipophilic substance to be determined, said complex liquid or host molecule being attached by a direct covalent bond or heteropolar bond or hydrogen bridge or hydrophobic bond to a chormophore or containing a chromo-phore in the form of an inclusion complex, whereafter the change of extinction or the wavelength displacement is measured; the present invention also provides a reagent for carrying out this process, as well as a test kit containing the reagent.
Description
The present invention is concerned with a process and a reagent for the determination of ions and of polar and/or lipophilic substances in fluids.
The qualitative and/or quantitative determination of individual ions in physiological fluids is of great importance for diagnosis, therapy and monitoring in the field of medicine, as well as in the control of chemical and micro-biological processes. In clinical diagnosis, chemical processes have hitherto been described with dyestuffs, as well as spectrophotometric processes and also methods using ion-selective electrodes.
In the case of flame emission analysis (flame photo-metry), the intensity of the light emitted by excited atoms is measured photoelectrically at the wavelength corresponding to the element.
In contradistinction thereto, according to the process of atom absorption spectroscopy, non-e~cited atoms are used, the concentration of which lies some factors of ten higher~ The advantages of these methods are the greater sensitivity, as well as the absence of spectral interference, Disadvantages in comparison with emission analysis include the relatively high expense of the apparatus, the very large sample volumes needed (2 ml.), the low sample frequency, as well as the separate and sequential determination of both parameters. Although this process is regarded as being a reference method for the determination of sodium and potassium nevertheless, it remains limited to medical-scientific laboratories.
For the simple and rapid determination of ion con-centrations or activities in aqueous solutions, ion-sensitive electrodes (ISE) can be used which, in comparison with the !
-- 1 -- : ~, 2B~
spectrophotometric process, require less expensive apparatus.
Disadvantages of these ion-selective electrodes include the relatively long and, in some cases, varying response time of the electrodes and the blocking up of the electrode sensers by high and low molecular weight substances present in the sample so that these only have relatively low stabilities.
The present invention seeks to provide a rapid and dependable process, which can be carried out not only in a liquid test but also by means of test strips, for the deter-mination of ions and of polar and/or lipophilic substances in fluids and especially in biological fluids, for ex~mple, blood serum.
Thus, according to the present invention, there is provided a process for the determination of an entity selected from the group consisting of ions, polar substances and lipo-philic substances, in fluids, wherein the entity to be deter-mined is allowed to act upon a complex ~igand or host molecule which is selective towards the entity to be determined, the complex ligand or host molecule being associated with at least one chromophore, whereafter a change of extinction or wave-length displacement is measured.
The present invention also provides a reagent for the determination of an entity selected from the group con-~
sisting of ions-, polar substances and lipophilic substances, in liquids, which contain at least one complex ligand or host molecule selective towards the entity, and associated with at least one chromophore.
Furthermore, the present invention provides a test kit comprising a reagent of the invention.
The entity to be determined is an ion, molecule or particle and may be herein referred to as a guest ion, guest molecule or guest particle.
34~
The complex ligand or host molecule may be associated with the at least one chromophore in a number of ways, includ-ing attachment to the chromophore by a direct covalent bond or heteropolar bond or hydrogen bridge or hydrophobic bond:
or the complex ligand or host molecula may contain the chromo-phore in the form of an inclusion complex.
There may be one or more selective complex ligands or host molecules, and these may be of the same or different type. Similarly there may be one or more chromophores of the same or different type.
The entity i9 allowed to act upon the complex ligand or host molecule in the sense that the liquid contain-ing the entity for determination is brought into contact with the complex ligand or host molecule and its associated at least one chromophore such that the absorption spectrum of the chromophore is modified by the interaction of the entity and the complex ligand or host molecule.
The selective complex ligand or selective host molecule is suitably a cyclic or acyclic medio- or macro-molecular compound which, with regard to the ion to bedetected or with regard to the polar and/or lipophilic substance to be detected is already present as a complex former or as a host molecule or, in the presence thereof, assumes the structure necessary for the complex formation or adduct formation in the form of a host-guest exchange action.
The polar ranges are hereby extended towards this in the presence of the ion.
The ring size and structure of the host molecule provide the selectivity in dependence upon the effective dia-meter of the specific polar or hydrophobic character of thecomplexing guest ion or guest molecule.
25~
In the case of complex formation, the ligand frequently changes i~s conformation and, as a rule, it is fixed, complex formation and comple~ dissociation thereby being influenced.
As complex ligands or host molecules there can be used compounds of the oligoether, polyether, oligo ester, polyester, oligoamide and polyamide types. Examples of suit-able compounds include crown ethers, cryptands, podands and derivatives thereof, as well as cyclic peptides and peptides which, in the presence of the ion to be determined or of the polar substance, assume the secondary, tertiary or quaternary structure necessary for the complex formation. Furthermore, there can be used tetrahydrofuran-containing, ester-bonded`
macrolides and analogous compounds which regulate or can regulate the transport in biological systems. There can also be used pure hydrocarbon structures, for example, lipo-philic host molecules ("lipophilic hollow spaces"), for example, cyclodextrins and cyclophanes, particularly where the entity to be determined is a lipophilic substance. Com-binations of the above functions are also possible.
The derivatives of the complex ligands or hostmolecules can possess bridges or chains which can contain oligo- or polyethylene glycol groupings or other hetero atom-containing groupings.
The chromophore attached to the complex ligand or host molecule as by a covalent, heteropolar or hydrophobic bond or by a hydrogen bridge is suitably a dyestuff or fluore-scent dyestuff or a chromogen, the absorption spectrum of which changes due to a reciprocal action, such as charge displace-ment and disturbance of the mesomerism either in the base and/or excited state by the guest ion or molecule. The dyestuffs used can be, for example, those of the polyene, meri~uinoid, quinone, azo (for example, methyl orange and methyl red), pyr-role, merocyanine, indigo, indophenol, stilbene, azomethine, anthraquinone, naphthoquinone, cyanine, phthalein, polymethine and alizarine types.
The chromophore contained in the complex ligand or host molecule in the form o~ an inclusion complex is ,suitably an acid dyestuff or a salt thereof, for example, a lithium, sodium, potassium, ammonium, calcium, alkylammonium or magnesium salt. The acid dyestuff may suitably contain a carboxylate, sulphonate, phenolate or thiophenolate grouping, as the acid grouping. The ions to be detected may be cations or anions. Cations which may be detected include, in parti-cular, alkali metal ions, for example, lithium, sodium or potassium ions, ammonium ions, alkaline earth metal ions, for example, magnesium or calcium ions; or other metal ions, including hea~y metal ions, for example, iron, zinc, copper, cobalt, nickel, molybdenum and chromium ions, In addition, there can be detected organic ions, for example, oligoalkyl-ammonium ions, phosphonium ions, guanidine ions and choline ions.
Anions which can be detected include, in particular,chloride, bromide, iodide, sulphate, nitrate, nitrite, phosphate, diphosphate, triphosphate, hydrogen phosphate and hydrogen carbonate ions.
~ eutral polar substances that may be detected include, by way of example, urea, thiourea, guanine, guanidine, uric acid, choline, creatinine, am~no acids and sugars.
Lipophilic guest molecules that may be detected include steroids, for example, cholesterol, and lipids, for example, triglycerides and lecithins.
The cation and anion concentration, as well as the concentrations of neutral guest particles can, in accordance with the invention be recognised qualitatively and quantit-atively by colour effects, i.e., photometrically, in a simple manner.
The selectivity of the complex ligand or host molecule can refer not only to a single, quite definite ion or substance to be detected but also to a group of ions or a group of substances. For this purpose, use can be made, for example, of chromophores, for example, dyestuffs, with several identical or different complex ligands or host molecules, for example, crown ethers of different hollow space size, such as are illustrated in the following formula:-~`0~
~o~\?~\c~ ~~~lm ~ ~ 1 N ~ ~
wherein 1, m and _ may be the same or different and are suit-ably 0, 1, 2, 3 or 4.
Furthermorej on to a complex ligand or a host molecule, such as the crown ether structure, there can be attached one or more identical or different chromophores, for example, dyestuffs.
The colour change in the determination of the invention may proceed, for example, according to the following mechanismu ~w~w=w~wl 3 ~ ~ w~ ~ Me ~ (II) .;
wherein Me ~ can be, for example, an alkali metal, alkaline earth metal, ammonium or heavy metal ion, and wherein the-mesomeric system is based upon the following e~uation (III), the weight and thus the energy content of which are usually different in the base and excited state:
O ~ ' ~`O O~~ ' - ~ N~3N=N~ ) O
O ~- 0~0~ , (III) N~D~,)= N-N ~,~= N~ (~) ~ , --m wherein m, is 0, 1, 2, 3 or 4.
In the chemistry of crown ethers, cryptands and podands there are many methods ~or linking comparatively large and average rings, as well as bi- and tricyclic systems with (CH2)n-, aryl- and hetero atom-containing structural elements.
Crown ethers, cryptands and podand molecules have the ability of forming stoichiometrical, crystalline complexes, as well as the selective or specific complexing of, for example, alkali metal, alkaline earth metal, ammonium and heavy metal ions, and also of neutral molecules. Numerous experimental results are available regardlng the phase transfer behaviour of the mentioned compounds.
~%~
Crown ethers, cryptands and podands are more particularly described in the review by F. Vogtle et al.
in Kontakte (E. Merck) 1/77, page 11; 2/77, page 16, 3/77, page 36; 2/78, page 16, 3/78, page 32, and 1/79, page 3.
In spite of the extensive investigations in the chemistry of the crown ethers, cryptands and podands, with regard, on the one hand, to syntheses and complexings, and, on the other hand, to the ion influencing of the colour, for example, in the case of mordant dyestuffs, complexometric indicators and porphyrin dyestuffs, hitherto nothing has been known regarding the combination of these two fields of investigation.
There is a series of publications regarding relevant bridgings in the case of cyclophanes and crown ethers, with the utilisation of the dilution principle and template effect.
Extensive knowledge is available regarding the synthesis and complexing behaviour of cyclic crown ethers and cryptands.
In addition, in particular, reactions have been successfully carried out on crown ether systems. Previous experience in this field enables, on the one hand, the synthesis of approp-riate crown ethers and cryptand structural units and, on the other hand, the introduction of oligoethylene glycol or other hetero atom-containing bridges.
A series of azo dyestuffs, as well as of di- and triphenylmethane dyestuffs, have been prepared from con-structional units of the following general formula (IV), in which Rl is a hydrogen atom and n is 1, 2 or 3, and of general formula (IV'). On these azo dyestuffs, there can be observed, depending upon the ring size, a selective displace-ment of the visible absorption maximum in a certain directionwhich is specific for each ion and which frequently involves a ~L2~34~
change of the molar extinction. The greatest of the spectral changes, such as AmaX, usually brings about the optimum appropriate cation in a particular crown ether ring within one Group of the Mendeleef Periodic System with regard to the radius.
Rl ~ ~ ~ (IV) f-o~o - - ..
n 0~ (IVI) (~ 0~ '' ' Various dyest~ffs systems have been provided with crown ether and cryptand units of variable ring size and structure. This hereby also involves the variation in the lipophilic properties or of the hydrophilic properties within the hollow space and on the periphery of the crown ether ring.
Both structural elements are to be joined to one another in such a manner that one or more donor hetero atoms of the ligand are simultaneously essential components of the chromo-phore. A comple~ed cation attacks a sensitive part of the chromophore more or less intensively and this specifically influences its absorption. This action on the absorbed system depends upon factors such as the size of the ion, its charge or charge density and solvation not only of the dye-stuff but also of the cation or anion, as well as the solvent.
~2~34~
The phenylaza-crown ethers of the general formula (IV) type are especially suitable because of their analogy with N,N-dimethylaniline and similar anilines which are often used in dyestuff chemistry. By means of azo coupling, dyestuffs of the general formula (IV) type can be prepared, the non-crowned analogues of which are known as light-fast textile dyestuffs, wherein, for example, _ is 1 and Rl is one of the following radicals:
02N- ~ CH=CH- , ~ N
The qualitative and/or quantitative determination of individual ions in physiological fluids is of great importance for diagnosis, therapy and monitoring in the field of medicine, as well as in the control of chemical and micro-biological processes. In clinical diagnosis, chemical processes have hitherto been described with dyestuffs, as well as spectrophotometric processes and also methods using ion-selective electrodes.
In the case of flame emission analysis (flame photo-metry), the intensity of the light emitted by excited atoms is measured photoelectrically at the wavelength corresponding to the element.
In contradistinction thereto, according to the process of atom absorption spectroscopy, non-e~cited atoms are used, the concentration of which lies some factors of ten higher~ The advantages of these methods are the greater sensitivity, as well as the absence of spectral interference, Disadvantages in comparison with emission analysis include the relatively high expense of the apparatus, the very large sample volumes needed (2 ml.), the low sample frequency, as well as the separate and sequential determination of both parameters. Although this process is regarded as being a reference method for the determination of sodium and potassium nevertheless, it remains limited to medical-scientific laboratories.
For the simple and rapid determination of ion con-centrations or activities in aqueous solutions, ion-sensitive electrodes (ISE) can be used which, in comparison with the !
-- 1 -- : ~, 2B~
spectrophotometric process, require less expensive apparatus.
Disadvantages of these ion-selective electrodes include the relatively long and, in some cases, varying response time of the electrodes and the blocking up of the electrode sensers by high and low molecular weight substances present in the sample so that these only have relatively low stabilities.
The present invention seeks to provide a rapid and dependable process, which can be carried out not only in a liquid test but also by means of test strips, for the deter-mination of ions and of polar and/or lipophilic substances in fluids and especially in biological fluids, for ex~mple, blood serum.
Thus, according to the present invention, there is provided a process for the determination of an entity selected from the group consisting of ions, polar substances and lipo-philic substances, in fluids, wherein the entity to be deter-mined is allowed to act upon a complex ~igand or host molecule which is selective towards the entity to be determined, the complex ligand or host molecule being associated with at least one chromophore, whereafter a change of extinction or wave-length displacement is measured.
The present invention also provides a reagent for the determination of an entity selected from the group con-~
sisting of ions-, polar substances and lipophilic substances, in liquids, which contain at least one complex ligand or host molecule selective towards the entity, and associated with at least one chromophore.
Furthermore, the present invention provides a test kit comprising a reagent of the invention.
The entity to be determined is an ion, molecule or particle and may be herein referred to as a guest ion, guest molecule or guest particle.
34~
The complex ligand or host molecule may be associated with the at least one chromophore in a number of ways, includ-ing attachment to the chromophore by a direct covalent bond or heteropolar bond or hydrogen bridge or hydrophobic bond:
or the complex ligand or host molecula may contain the chromo-phore in the form of an inclusion complex.
There may be one or more selective complex ligands or host molecules, and these may be of the same or different type. Similarly there may be one or more chromophores of the same or different type.
The entity i9 allowed to act upon the complex ligand or host molecule in the sense that the liquid contain-ing the entity for determination is brought into contact with the complex ligand or host molecule and its associated at least one chromophore such that the absorption spectrum of the chromophore is modified by the interaction of the entity and the complex ligand or host molecule.
The selective complex ligand or selective host molecule is suitably a cyclic or acyclic medio- or macro-molecular compound which, with regard to the ion to bedetected or with regard to the polar and/or lipophilic substance to be detected is already present as a complex former or as a host molecule or, in the presence thereof, assumes the structure necessary for the complex formation or adduct formation in the form of a host-guest exchange action.
The polar ranges are hereby extended towards this in the presence of the ion.
The ring size and structure of the host molecule provide the selectivity in dependence upon the effective dia-meter of the specific polar or hydrophobic character of thecomplexing guest ion or guest molecule.
25~
In the case of complex formation, the ligand frequently changes i~s conformation and, as a rule, it is fixed, complex formation and comple~ dissociation thereby being influenced.
As complex ligands or host molecules there can be used compounds of the oligoether, polyether, oligo ester, polyester, oligoamide and polyamide types. Examples of suit-able compounds include crown ethers, cryptands, podands and derivatives thereof, as well as cyclic peptides and peptides which, in the presence of the ion to be determined or of the polar substance, assume the secondary, tertiary or quaternary structure necessary for the complex formation. Furthermore, there can be used tetrahydrofuran-containing, ester-bonded`
macrolides and analogous compounds which regulate or can regulate the transport in biological systems. There can also be used pure hydrocarbon structures, for example, lipo-philic host molecules ("lipophilic hollow spaces"), for example, cyclodextrins and cyclophanes, particularly where the entity to be determined is a lipophilic substance. Com-binations of the above functions are also possible.
The derivatives of the complex ligands or hostmolecules can possess bridges or chains which can contain oligo- or polyethylene glycol groupings or other hetero atom-containing groupings.
The chromophore attached to the complex ligand or host molecule as by a covalent, heteropolar or hydrophobic bond or by a hydrogen bridge is suitably a dyestuff or fluore-scent dyestuff or a chromogen, the absorption spectrum of which changes due to a reciprocal action, such as charge displace-ment and disturbance of the mesomerism either in the base and/or excited state by the guest ion or molecule. The dyestuffs used can be, for example, those of the polyene, meri~uinoid, quinone, azo (for example, methyl orange and methyl red), pyr-role, merocyanine, indigo, indophenol, stilbene, azomethine, anthraquinone, naphthoquinone, cyanine, phthalein, polymethine and alizarine types.
The chromophore contained in the complex ligand or host molecule in the form o~ an inclusion complex is ,suitably an acid dyestuff or a salt thereof, for example, a lithium, sodium, potassium, ammonium, calcium, alkylammonium or magnesium salt. The acid dyestuff may suitably contain a carboxylate, sulphonate, phenolate or thiophenolate grouping, as the acid grouping. The ions to be detected may be cations or anions. Cations which may be detected include, in parti-cular, alkali metal ions, for example, lithium, sodium or potassium ions, ammonium ions, alkaline earth metal ions, for example, magnesium or calcium ions; or other metal ions, including hea~y metal ions, for example, iron, zinc, copper, cobalt, nickel, molybdenum and chromium ions, In addition, there can be detected organic ions, for example, oligoalkyl-ammonium ions, phosphonium ions, guanidine ions and choline ions.
Anions which can be detected include, in particular,chloride, bromide, iodide, sulphate, nitrate, nitrite, phosphate, diphosphate, triphosphate, hydrogen phosphate and hydrogen carbonate ions.
~ eutral polar substances that may be detected include, by way of example, urea, thiourea, guanine, guanidine, uric acid, choline, creatinine, am~no acids and sugars.
Lipophilic guest molecules that may be detected include steroids, for example, cholesterol, and lipids, for example, triglycerides and lecithins.
The cation and anion concentration, as well as the concentrations of neutral guest particles can, in accordance with the invention be recognised qualitatively and quantit-atively by colour effects, i.e., photometrically, in a simple manner.
The selectivity of the complex ligand or host molecule can refer not only to a single, quite definite ion or substance to be detected but also to a group of ions or a group of substances. For this purpose, use can be made, for example, of chromophores, for example, dyestuffs, with several identical or different complex ligands or host molecules, for example, crown ethers of different hollow space size, such as are illustrated in the following formula:-~`0~
~o~\?~\c~ ~~~lm ~ ~ 1 N ~ ~
wherein 1, m and _ may be the same or different and are suit-ably 0, 1, 2, 3 or 4.
Furthermorej on to a complex ligand or a host molecule, such as the crown ether structure, there can be attached one or more identical or different chromophores, for example, dyestuffs.
The colour change in the determination of the invention may proceed, for example, according to the following mechanismu ~w~w=w~wl 3 ~ ~ w~ ~ Me ~ (II) .;
wherein Me ~ can be, for example, an alkali metal, alkaline earth metal, ammonium or heavy metal ion, and wherein the-mesomeric system is based upon the following e~uation (III), the weight and thus the energy content of which are usually different in the base and excited state:
O ~ ' ~`O O~~ ' - ~ N~3N=N~ ) O
O ~- 0~0~ , (III) N~D~,)= N-N ~,~= N~ (~) ~ , --m wherein m, is 0, 1, 2, 3 or 4.
In the chemistry of crown ethers, cryptands and podands there are many methods ~or linking comparatively large and average rings, as well as bi- and tricyclic systems with (CH2)n-, aryl- and hetero atom-containing structural elements.
Crown ethers, cryptands and podand molecules have the ability of forming stoichiometrical, crystalline complexes, as well as the selective or specific complexing of, for example, alkali metal, alkaline earth metal, ammonium and heavy metal ions, and also of neutral molecules. Numerous experimental results are available regardlng the phase transfer behaviour of the mentioned compounds.
~%~
Crown ethers, cryptands and podands are more particularly described in the review by F. Vogtle et al.
in Kontakte (E. Merck) 1/77, page 11; 2/77, page 16, 3/77, page 36; 2/78, page 16, 3/78, page 32, and 1/79, page 3.
In spite of the extensive investigations in the chemistry of the crown ethers, cryptands and podands, with regard, on the one hand, to syntheses and complexings, and, on the other hand, to the ion influencing of the colour, for example, in the case of mordant dyestuffs, complexometric indicators and porphyrin dyestuffs, hitherto nothing has been known regarding the combination of these two fields of investigation.
There is a series of publications regarding relevant bridgings in the case of cyclophanes and crown ethers, with the utilisation of the dilution principle and template effect.
Extensive knowledge is available regarding the synthesis and complexing behaviour of cyclic crown ethers and cryptands.
In addition, in particular, reactions have been successfully carried out on crown ether systems. Previous experience in this field enables, on the one hand, the synthesis of approp-riate crown ethers and cryptand structural units and, on the other hand, the introduction of oligoethylene glycol or other hetero atom-containing bridges.
A series of azo dyestuffs, as well as of di- and triphenylmethane dyestuffs, have been prepared from con-structional units of the following general formula (IV), in which Rl is a hydrogen atom and n is 1, 2 or 3, and of general formula (IV'). On these azo dyestuffs, there can be observed, depending upon the ring size, a selective displace-ment of the visible absorption maximum in a certain directionwhich is specific for each ion and which frequently involves a ~L2~34~
change of the molar extinction. The greatest of the spectral changes, such as AmaX, usually brings about the optimum appropriate cation in a particular crown ether ring within one Group of the Mendeleef Periodic System with regard to the radius.
Rl ~ ~ ~ (IV) f-o~o - - ..
n 0~ (IVI) (~ 0~ '' ' Various dyest~ffs systems have been provided with crown ether and cryptand units of variable ring size and structure. This hereby also involves the variation in the lipophilic properties or of the hydrophilic properties within the hollow space and on the periphery of the crown ether ring.
Both structural elements are to be joined to one another in such a manner that one or more donor hetero atoms of the ligand are simultaneously essential components of the chromo-phore. A comple~ed cation attacks a sensitive part of the chromophore more or less intensively and this specifically influences its absorption. This action on the absorbed system depends upon factors such as the size of the ion, its charge or charge density and solvation not only of the dye-stuff but also of the cation or anion, as well as the solvent.
~2~34~
The phenylaza-crown ethers of the general formula (IV) type are especially suitable because of their analogy with N,N-dimethylaniline and similar anilines which are often used in dyestuff chemistry. By means of azo coupling, dyestuffs of the general formula (IV) type can be prepared, the non-crowned analogues of which are known as light-fast textile dyestuffs, wherein, for example, _ is 1 and Rl is one of the following radicals:
02N- ~ CH=CH- , ~ N
2 ~1 ~ ~=N-`S
or NO~
The auxochromic crown ether nitrogen here permits a hypsochromic displacement of the absorption maximum in the case of the exchange reaction with the guest ion since its free electron pair, after complexing has taken place, is not available or is only partly available for mesometry, depending upon the nature of the complexed ion, for example, the charge~
A further increase of the ion selectivity can be achieved by the partial reinforcement of the crown ether ring, for example;
in the phenylaza-benzo-crown ethers of the general formula (V) type, in which n is 1, ~ or 3, which can be used, for example, as azo coupling partners and for similar reactions.
11~2~34~.
~ ~1 ~ ~_ O ~ n There are hereby especially preferred the chromo~
phore systems in which an easy influencability by a complexed ion is to be expected, for example, those with a high solvatochromy, for example, indophenols of the general formula (IV) type, in which Rl is a radical of the formula:
~ ~r . , .
which`can be prepared via compounds of general formula (IV), in which Rl is a nitro or primary amino group. The last : step includes an oxidative coupling with phenols. The analogous CH compounds of general formula (IV), in which Rl -is the radical:
CH
4~L
can be prepared from the corresponding aldehydes, in which Rl is -CHO, whlch can be obtained, for example, vla a Vilsmeier reaction.
Furthermore, stilbenes and merocyanines and possibly aza analogues of general formula (IV) are suitable, wherein n is, for example, l and Rl is, for example, one of the following radicals:
, ~ CH=CH-CH
4 ~ CH=CH- or R4- ~ N=CH-R4 being an alkyl radical containing, for example, up to 5 carbon atoms and preferably being a methyl or ethyl radical.
The attachment of dyestufs in the above manner to cryptand systems leads to the achievement of a comparatively high complex stability, especially in an aqueous medium, and to a comparatively high selectivity. In this case, it is especially advantageous to couple reactive diazonium salts with benzo-cryptade~ of the general formula:
~-, wherein R2 can be, for exhmple, a radical of the formula:
~2 ~ ~ or 2~ ~ N=~-2 S ~=N- N2 Benzocrown ethers of the general formula:
~~O ~
~VII) <_O
have proved to be model substances for the synthesis which also give conclusions regarding the behavi.ollr to be expected with regard to ions. The variation of the hollow space size requires benzocryptands, such as are n~3eded for compounds of general formula (VI), but which, however, are shortened 10 : or lengthened by one or more oxyethylene units.
Furthermore, cryptands of the following type:
N
~- N
\/
are also suitable, In a similar manner, there can also be prepared those of the following type:
~ N
`~
O ~ .
which, similarly to the simple phenylaza-crown ethers, can, by reaction in the 4-position of the aniline moiety, be reacted to give dyestuff cryptands of, for example, the following general formula:
N = N ~
(IXa) ~O O~
N ~
W ~ R3
or NO~
The auxochromic crown ether nitrogen here permits a hypsochromic displacement of the absorption maximum in the case of the exchange reaction with the guest ion since its free electron pair, after complexing has taken place, is not available or is only partly available for mesometry, depending upon the nature of the complexed ion, for example, the charge~
A further increase of the ion selectivity can be achieved by the partial reinforcement of the crown ether ring, for example;
in the phenylaza-benzo-crown ethers of the general formula (V) type, in which n is 1, ~ or 3, which can be used, for example, as azo coupling partners and for similar reactions.
11~2~34~.
~ ~1 ~ ~_ O ~ n There are hereby especially preferred the chromo~
phore systems in which an easy influencability by a complexed ion is to be expected, for example, those with a high solvatochromy, for example, indophenols of the general formula (IV) type, in which Rl is a radical of the formula:
~ ~r . , .
which`can be prepared via compounds of general formula (IV), in which Rl is a nitro or primary amino group. The last : step includes an oxidative coupling with phenols. The analogous CH compounds of general formula (IV), in which Rl -is the radical:
CH
4~L
can be prepared from the corresponding aldehydes, in which Rl is -CHO, whlch can be obtained, for example, vla a Vilsmeier reaction.
Furthermore, stilbenes and merocyanines and possibly aza analogues of general formula (IV) are suitable, wherein n is, for example, l and Rl is, for example, one of the following radicals:
, ~ CH=CH-CH
4 ~ CH=CH- or R4- ~ N=CH-R4 being an alkyl radical containing, for example, up to 5 carbon atoms and preferably being a methyl or ethyl radical.
The attachment of dyestufs in the above manner to cryptand systems leads to the achievement of a comparatively high complex stability, especially in an aqueous medium, and to a comparatively high selectivity. In this case, it is especially advantageous to couple reactive diazonium salts with benzo-cryptade~ of the general formula:
~-, wherein R2 can be, for exhmple, a radical of the formula:
~2 ~ ~ or 2~ ~ N=~-2 S ~=N- N2 Benzocrown ethers of the general formula:
~~O ~
~VII) <_O
have proved to be model substances for the synthesis which also give conclusions regarding the behavi.ollr to be expected with regard to ions. The variation of the hollow space size requires benzocryptands, such as are n~3eded for compounds of general formula (VI), but which, however, are shortened 10 : or lengthened by one or more oxyethylene units.
Furthermore, cryptands of the following type:
N
~- N
\/
are also suitable, In a similar manner, there can also be prepared those of the following type:
~ N
`~
O ~ .
which, similarly to the simple phenylaza-crown ethers, can, by reaction in the 4-position of the aniline moiety, be reacted to give dyestuff cryptands of, for example, the following general formula:
N = N ~
(IXa) ~O O~
N ~
W ~ R3
3 (IXb) ~,J
~2~
wherein R3 is a nitro, cyano, sulphonium or like group.
Besides the previously mentioned types, in which a mesomery influencing takes place, for example, by a dis-turbance due to incorporated ions on a hetero atom, an attack can also take place on the antiauxochrome or on an azo bridge according to the following general formulae:
~/~ 0~ ~,N
R6~ G~C~C~L Me~ (X~
~~
wherèin each R6 is a lower alkyl group of 1 to 6, preferably 1 to 4 carbon atoms, or the two R6 groups together with the nitrogen atom to which they are attached form a ring, and R7 and R8, which can be the same or dif:Eerent, are hydrogen atoms or nitro, cyano or dialkylamino groups in which the alkyl groups contain 1 to 6 and preferably 1 to 4 carbon atoms.
Examples of fluorescent dyestuff crown ethers are those of the formula:-~ C ~ (XIa) ~C02~
Further examples of dyestuff crown ethers are thoseof the general formula (XIb) (wherein X is a nitrogen atom or -CH=, Y is -CH2-, -NH- or an oxygen or sulphur atom and n is 1, 2 or 3), such as methylene blue; crown ethers of the general formulae (XIc) and (XId) (azomethine dyestuff crown ethers), wherein Rg is a lower alkyl radical of l to 6 and preferably 1 to 4 carbon atoms, and A ~3 is a conventional anion' crown ethers of the formula (XIe) (anthracilin crown ethers), crown ethers of the formula (XIf) (alizarin crown ethers) and crown ethers of the formula (XIg) (cyanine type). An example of - a lipophilic hollow space is the compound of general formula (XII), in which n is a whole number of from 1 to 6 and Rlo is a hydrogen atom or a lower alkyl of L to 6 and preferably 1 to 4 carbon atoms or an aryl radical suitably a phenyl or naphthyl radical.
o~ (~Ib) ~n ~E r .
N ~`~ - ~ N ~
(XIc) .. ~ O
\J \ /
~O 0~ ~XId ~
~ + D\D ~ ~3 RgA Rg _>~ ' '~
. O
R ~ ~ ~ ~ ~ ~ 3 ~l2)n As fluorescent dyestuffs, those of the phthalein group (general formula (XIa)) are especially preferred, the crown ether rings thereby being applied to the chromophore at the places which are especially sensitively influenced in the case of complexing. In the case of such fluorescent dye-stuffs, a stronger disturbance of the colour or of the fluore-scence (phosphorescence) of the chromophore is present due to the complexing. By adjustment of the crown ether hollow space to certain ions, such exchange actions and effects can be selectively adjusted to certain kinds of ions. Apart from the alkali metal and alkaline earth metal ions, the heavy metal ions, as well as ammonium ions and other organic onium ions, for example, the phosphonium ions, play a part. Thus, for example, in this case amntonium group-containing disinfection agents can be detected.
By the introduction of dyestuff crown ethers (or, vice versa, of ions) into fibres or synthetic resin films or the like, by means of a dyeing process, which can correspond to mordanting, not only can dyestuffs be fixed on to textiles and the like but also modified dyeing processes and colour nuances can be achieved. The binding is thereby not ionic and not covalent but rather takes place via a crown ether host-guest reciprocal action, i.e., ion-dlpole reciprocal action, it can be made reversible and thus can be analytically evaluated.
In particular, use can also be made of anthra-quinones with functional groups, -for example, sulphonic acid groups, or of naphthoquinone-sulphonic acids which have previously been used or are even today still used in dyeing.
According to the present invention, they can be ion-selectively modified and thus serve for the detection of particular ions~ They can also be applied or bound to paper or fibres, as well as to polymers.
In a further aspect the invention provides a test device in which the reagent of the invention is applied to an inert substrate. r~he reagent may be formed as a film on the substrate or impregnated in a porous substrate. Suit-able substrates include solid carriers, for example, paper, synthetic resin films, glass, aluminium oxide, silicon oxide, natural and synthetic fibres and textile materials, and metals.
In the case of all dyestuffs and especially of the cyanine dyestuffs, reference is made to the appropriate organic chemical textbooks and dyestuff reference books A
very suitable cyanine dyestuff is Konig's salt (formula (XIg)) which can easily be prepared from pyridine and N-methylaniline.
~2~
wherein R3 is a nitro, cyano, sulphonium or like group.
Besides the previously mentioned types, in which a mesomery influencing takes place, for example, by a dis-turbance due to incorporated ions on a hetero atom, an attack can also take place on the antiauxochrome or on an azo bridge according to the following general formulae:
~/~ 0~ ~,N
R6~ G~C~C~L Me~ (X~
~~
wherèin each R6 is a lower alkyl group of 1 to 6, preferably 1 to 4 carbon atoms, or the two R6 groups together with the nitrogen atom to which they are attached form a ring, and R7 and R8, which can be the same or dif:Eerent, are hydrogen atoms or nitro, cyano or dialkylamino groups in which the alkyl groups contain 1 to 6 and preferably 1 to 4 carbon atoms.
Examples of fluorescent dyestuff crown ethers are those of the formula:-~ C ~ (XIa) ~C02~
Further examples of dyestuff crown ethers are thoseof the general formula (XIb) (wherein X is a nitrogen atom or -CH=, Y is -CH2-, -NH- or an oxygen or sulphur atom and n is 1, 2 or 3), such as methylene blue; crown ethers of the general formulae (XIc) and (XId) (azomethine dyestuff crown ethers), wherein Rg is a lower alkyl radical of l to 6 and preferably 1 to 4 carbon atoms, and A ~3 is a conventional anion' crown ethers of the formula (XIe) (anthracilin crown ethers), crown ethers of the formula (XIf) (alizarin crown ethers) and crown ethers of the formula (XIg) (cyanine type). An example of - a lipophilic hollow space is the compound of general formula (XII), in which n is a whole number of from 1 to 6 and Rlo is a hydrogen atom or a lower alkyl of L to 6 and preferably 1 to 4 carbon atoms or an aryl radical suitably a phenyl or naphthyl radical.
o~ (~Ib) ~n ~E r .
N ~`~ - ~ N ~
(XIc) .. ~ O
\J \ /
~O 0~ ~XId ~
~ + D\D ~ ~3 RgA Rg _>~ ' '~
. O
R ~ ~ ~ ~ ~ ~ 3 ~l2)n As fluorescent dyestuffs, those of the phthalein group (general formula (XIa)) are especially preferred, the crown ether rings thereby being applied to the chromophore at the places which are especially sensitively influenced in the case of complexing. In the case of such fluorescent dye-stuffs, a stronger disturbance of the colour or of the fluore-scence (phosphorescence) of the chromophore is present due to the complexing. By adjustment of the crown ether hollow space to certain ions, such exchange actions and effects can be selectively adjusted to certain kinds of ions. Apart from the alkali metal and alkaline earth metal ions, the heavy metal ions, as well as ammonium ions and other organic onium ions, for example, the phosphonium ions, play a part. Thus, for example, in this case amntonium group-containing disinfection agents can be detected.
By the introduction of dyestuff crown ethers (or, vice versa, of ions) into fibres or synthetic resin films or the like, by means of a dyeing process, which can correspond to mordanting, not only can dyestuffs be fixed on to textiles and the like but also modified dyeing processes and colour nuances can be achieved. The binding is thereby not ionic and not covalent but rather takes place via a crown ether host-guest reciprocal action, i.e., ion-dlpole reciprocal action, it can be made reversible and thus can be analytically evaluated.
In particular, use can also be made of anthra-quinones with functional groups, -for example, sulphonic acid groups, or of naphthoquinone-sulphonic acids which have previously been used or are even today still used in dyeing.
According to the present invention, they can be ion-selectively modified and thus serve for the detection of particular ions~ They can also be applied or bound to paper or fibres, as well as to polymers.
In a further aspect the invention provides a test device in which the reagent of the invention is applied to an inert substrate. r~he reagent may be formed as a film on the substrate or impregnated in a porous substrate. Suit-able substrates include solid carriers, for example, paper, synthetic resin films, glass, aluminium oxide, silicon oxide, natural and synthetic fibres and textile materials, and metals.
In the case of all dyestuffs and especially of the cyanine dyestuffs, reference is made to the appropriate organic chemical textbooks and dyestuff reference books A
very suitable cyanine dyestuff is Konig's salt (formula (XIg)) which can easily be prepared from pyridine and N-methylaniline.
4~
Konig's salt can be modified by the attachment of crown ethers in such a manner that, in the case of complex formation with alkali metal or alkaline earth metal or heavy metal ions, colour changes or colour effects occur which can be used for the detection or concentration determination of these ions, even in the case of the simultaneous presence of o-ther ions.
One possibility for the modification of cyanine dyestuffs of the Konig's salt type consists in the bridging of the two nitrogen centres by a crown ether-like cyclic or open-chained crown ether unit with donor groups according to the following general formula:- -A~ o 0 0 ~ (XIII), L J m wherein m is 1, 2 or 3 and A ~ is an anion.
In this ~ay, the chromophore is additionally dis-turbed in the case of the host-guest reciprocal action, i.e., in the case of nesting in of the cation, due to the positive charge of the cation. Due to the crown ether structure attached directly on to the sensitive part (electron cloud~
of the chromophore, the cation is attracted and-fi~ed and is able to change this chromophore system by the attraction of electrons from the loose ~-electron cloud, which brings about the colour effect. By means of shorter or longer crown ether bridges, which can be modified by various hetero atoms or by ~'#.
.
, .
the incorporation of rigid aromatic structural elements, for example, pyridine rings or the like, these dyestuffs can be influenced in the desired manner in the case of the com-plexing.
Another manner of preparing crown ethers with a cyanine structure of the Konig's salt type consists in start- -ing from an aniline provided in the ~-position with an aza crown ether ring and, in the case of the complexing of the lone electron pair on the ~-amino nitrogen of the crown ether ring, sensitively to disturb by the nesting in of the ion so that this lone electron pair is only available to a limited extent for mesomerism, which forces a colour change.
The present invention also includes the use of complex ligands in the form of a cyclic peptide or of a peptide which, in the presence of the :ion or of the polar substance, assumes the necessary secondary, tertiary or quaternary structure. Thus, for examp:Le, natural ionophores, such as valinomycin, nonactin, gramacidin and similar peptidès;
can also be employed for the ion-selective colour reactions or colour tests according to the present invention. In this case, the procedure can be as follows: into valinomycin there is incorporated a dyestuff salt, for example, 2,4-dinitro-phenylhydrazonium chloride, sodium picrate or a similar dye-stuff~ The valinomycin-dyestuff complex generally has a different colour or absorption maximum from that of the free dyestuff. In a second step, to this valinomycin-dyestuff complex there is added the salt solution, the concentration of which is to be tested, the potassium-specific valinomycin thereby complexing the potassium ions present in the solution, the dyestuff being forced out of the valinomycin hollow spaces and another colour or absorption spectrum now again being displayed in the solution.
In this way, the potassium specificity of valino-mycin can be utilised ~or colour tests. As a simple dyestuff system which can be used for incorporation into or adding on to valinomycin and subsequent expulsion with potassium ions, there can be used, in particular, hydrazinium salts with various attached mesomeric systems, for example, azo functions. Dyestuff cations are used which are sterically as little demanding as possible and which form with valinomycin a weak but still sufficiently stable complex which n~st also not be too strong so that it can thereafter again be quantitatively broked down with potassium ions, even in low concentration.
Analogous experiments with crown ethers, for example, the crown ether [18] crown-6, which thus has a structure which is comparable with that of valinomycin and nonactin, permits the recognition of marked colour ef~ects or changes of the long wavelength absorption of the dye-- stuff in the state of being bound on to the crown ether and in the free state, The use not only of cationic dyestuff salts, for example, 2,4-dinitrophenylhydrazonium ions, as well as of anionic colour carriers, for example picrate ions, offers a plenitude of variation and application possibilities, the choice of which can be exactly "tailored" from case to case, iOe., to the components employed~
Another dyestuff which is preferred is the azo compound of the formula:-2 ~ N = N ~ N02.HCl (XI~) 2~
As examples of neutral guest molecules which, according to the present invention, can be selectively detected, there may be mentioned, in particular, urea, thiourea, ammonium salts, for example, choline, guanine, guanidine, uric acid, creatinine, amino acids and sugars. These substances may he determined on the basis of their colour effects, not only qualitatively but also quantitatively. For the quantitative determination of urea in aqueous solutions, a quinoline ligand can be synthesised which can be used not only for urea but also for thiourea ~see Tetrahedron Letters, 3, 309-312/1978).
The process of the invention with the use of the novel dyestuff systems is of great interest since it readily permits a plenitude of measurements and calculations which extend, for example, to the ion selectivity, caused by the complex constants with regard to various ions and the influence of ions of particular charge density on the chromo-phore system of the most va~ied types of dyestuffs with regard to extinction ànd absorption properties. In many cases, pre-liminary calculations with molecule orbi~al methods arepossible.
Another structural variant of the intramolecular ionophore/chromophore combination of the (XV) or (XVI) type is shown in another manner influencing selectivity: the cations fixed in the crown ether hollow space are additionally co-ordinated by the donor centre present on a comparatively short or comparatively long side chain, which can be a phenolic or CH20 ~ group or C00 ~3 or S03 ~3. Two influences are here combined: the negative charge of the side chain bonds the cation electrostatically more strongly than would be possible by a crown ether ring because of the ion-dipole reciprocal action. However, the cation must, nevertheless, pass into the crown ether ring, i.e., it is there tested and selected for itssize. Overall, higher complex constants are experimentally observed on the basis of the stronger bonding of the cation to the ligands, a higher selectivity of the cation embedded in the crown ether ring, as well as, as a result thereoE, a strong influencing of the chromophore system because of the favourable position of the equilibrium in comparison with the previously described ligands and a special selectivity towards cations with high charge density, for example, calcium ions. Such ligand systems, after optimisation to certain cations, can possibly be used for the speci-Eic differentiation of simple/plural charged cations:-H3C ~` ~ (XV) -m ~C~ - '~
2 ~ ~ N ~ (XVI) [CH2 ~COO (~7 .
~ - 24 -H3C 0 /~\ A ~ CH3 E3C ~ ~ ~ r-~ ~ ~ N ~ N
CH -~ ~ 2 The process according to the present invention provides a series of novel and practical uses:
a) With the help of the crown ether dyestuffs, indicators and probes can be developed, for example, for the detection of the phase trans~er of salts, for the study of ion trans-port by lipophilic media, for example, synthetic and biological membranes, i.e., for investigations in connection with membranological research from a physiological or pathological 0 ~point of view.
b) Another use is the possibility of ion-selectively colouring tissue sections. A possibly quantitative evaluation hereby permits conclusions regarding the nature and concent-ration of ions which are made visible by the displacement of AmaX and extinction.
c) The use of chromophore cryptand systems ensures, especially in aqueous media, not only a substantially higher complex stability but also an improved selectivity. Besides the qualitative and quantitative photometric detection o-f the alkali metals and alkaline earth metals, as well as of the ammonium ions, with the help of host chromophores of various hollow space size in solutions, for example, in blood or sera and other body fluids, these can also be identified on carrier materials. The use as spray reagents for ion chromatograms or silicone prints in the field of medical diagnosis is hereby possible. In the manner of thermography, either by spraying areas or tissue or their imprints with ion-selective dye-stuffs and fluorescent dyestuffs, physiological or pathological salt concentrations can be made visible. From this there follows the possibility of differentiating disease.
d) From the knowledge of the complexing of organic ammonium and guanidinium salts, as well as of neutral mole-cules, for example, urea and CH-acid compounds, it is, according to the present invention, now possible to carry out detection with appropriate host chromophores. The useful-ness from a medicinal or biochemical point o~ view hereby lies in the specific photometric detection of certain sub-stances which are peculiar to the body and the kinetic monitoring of enzymatic processes, Having regard thereto, there can be considered the attachment of lipophilic hollow spaces in the form of endolipiphilic or endohydrophobic macrocyclic compounds with exohydrophilic or exopolarophilic molecular peripheries on sensitive i.e~ chromophore-influencing, positions of one of the dyestuffs which permit the enveloping of an appropriate neutral particle and, probably due to hydrogen bridges, bring about a change of the absorption or extinction.
The invention is further illustrated by reference to the accompanying drawings, referred to more particularly in the examples in which:
FIGURE 1 shows graphically the absorption spectra of 2,4-dinitro-4'-(4,7,10,13-tetraoxa-1-azocyclopentadec-l-yl)-stilbene with various metal ions, according to Example 3, FIGURE 2 shows graphically the absorption spectra of 2,4-dinitro-4'-(4,7,10,13,16-pentaoxa-1-azacyclooctadec-l-yl)-stilbene and its com-plexes with ammonium ions and various metal ions, according to Example 6, FIGURE 3 illustrates graphically the absorption spectra of a blue dyestuff and its com-plexes with various metal ions, according to Example 7, FIGURE 4 illustrates graphically the absorption spectra of a dyestuff and its complexes with various metal ions, according to Example 8, FIGURE S illustrates graphically the absorption spectra of 1,8-bis[l-(4-dimethylamino-phenylimino)-~-benzoquin-3-oxy~-3,6-dioxaoctane and its complexes with various metal ions, according to Example 9, and FIGURE 6 illustrates graphically the absorption spectra of a dyestuff and its complexes .-with various metal ions, according to .
. Example 10.
: The following Examples are given for the purpose of ~:~ 20 illustrating the present invention:
: Example 1 .
Photometric determination of potassium with an ion-selective dyestuff which contains the chromophore covalently bound via a mesomeric system An ion-selective dye.stuff of general formula (IV')`
(n = 1) is dissolved in chloroform and shaken up with a solution which contains potassium ions. The dyestuff com-plexed with potassium passes over into the aqueous phase and can be determined quantitatively at 600nm in a photometer, depending upon the activity of the potassium ions present.
i~
The dyestuff (5-nitro-1,3-thiazole-2-azo-[3-(12- -hydroxy-1,4,7,10-tetraoxadodecyl]-4-hydroxybenzene) can be prepared by reacting 2-amino~5-nitrothiazole in 85% phosphoric acid with sodium nitrite and subsequently adding benzo[l5] crown-
Konig's salt can be modified by the attachment of crown ethers in such a manner that, in the case of complex formation with alkali metal or alkaline earth metal or heavy metal ions, colour changes or colour effects occur which can be used for the detection or concentration determination of these ions, even in the case of the simultaneous presence of o-ther ions.
One possibility for the modification of cyanine dyestuffs of the Konig's salt type consists in the bridging of the two nitrogen centres by a crown ether-like cyclic or open-chained crown ether unit with donor groups according to the following general formula:- -A~ o 0 0 ~ (XIII), L J m wherein m is 1, 2 or 3 and A ~ is an anion.
In this ~ay, the chromophore is additionally dis-turbed in the case of the host-guest reciprocal action, i.e., in the case of nesting in of the cation, due to the positive charge of the cation. Due to the crown ether structure attached directly on to the sensitive part (electron cloud~
of the chromophore, the cation is attracted and-fi~ed and is able to change this chromophore system by the attraction of electrons from the loose ~-electron cloud, which brings about the colour effect. By means of shorter or longer crown ether bridges, which can be modified by various hetero atoms or by ~'#.
.
, .
the incorporation of rigid aromatic structural elements, for example, pyridine rings or the like, these dyestuffs can be influenced in the desired manner in the case of the com-plexing.
Another manner of preparing crown ethers with a cyanine structure of the Konig's salt type consists in start- -ing from an aniline provided in the ~-position with an aza crown ether ring and, in the case of the complexing of the lone electron pair on the ~-amino nitrogen of the crown ether ring, sensitively to disturb by the nesting in of the ion so that this lone electron pair is only available to a limited extent for mesomerism, which forces a colour change.
The present invention also includes the use of complex ligands in the form of a cyclic peptide or of a peptide which, in the presence of the :ion or of the polar substance, assumes the necessary secondary, tertiary or quaternary structure. Thus, for examp:Le, natural ionophores, such as valinomycin, nonactin, gramacidin and similar peptidès;
can also be employed for the ion-selective colour reactions or colour tests according to the present invention. In this case, the procedure can be as follows: into valinomycin there is incorporated a dyestuff salt, for example, 2,4-dinitro-phenylhydrazonium chloride, sodium picrate or a similar dye-stuff~ The valinomycin-dyestuff complex generally has a different colour or absorption maximum from that of the free dyestuff. In a second step, to this valinomycin-dyestuff complex there is added the salt solution, the concentration of which is to be tested, the potassium-specific valinomycin thereby complexing the potassium ions present in the solution, the dyestuff being forced out of the valinomycin hollow spaces and another colour or absorption spectrum now again being displayed in the solution.
In this way, the potassium specificity of valino-mycin can be utilised ~or colour tests. As a simple dyestuff system which can be used for incorporation into or adding on to valinomycin and subsequent expulsion with potassium ions, there can be used, in particular, hydrazinium salts with various attached mesomeric systems, for example, azo functions. Dyestuff cations are used which are sterically as little demanding as possible and which form with valinomycin a weak but still sufficiently stable complex which n~st also not be too strong so that it can thereafter again be quantitatively broked down with potassium ions, even in low concentration.
Analogous experiments with crown ethers, for example, the crown ether [18] crown-6, which thus has a structure which is comparable with that of valinomycin and nonactin, permits the recognition of marked colour ef~ects or changes of the long wavelength absorption of the dye-- stuff in the state of being bound on to the crown ether and in the free state, The use not only of cationic dyestuff salts, for example, 2,4-dinitrophenylhydrazonium ions, as well as of anionic colour carriers, for example picrate ions, offers a plenitude of variation and application possibilities, the choice of which can be exactly "tailored" from case to case, iOe., to the components employed~
Another dyestuff which is preferred is the azo compound of the formula:-2 ~ N = N ~ N02.HCl (XI~) 2~
As examples of neutral guest molecules which, according to the present invention, can be selectively detected, there may be mentioned, in particular, urea, thiourea, ammonium salts, for example, choline, guanine, guanidine, uric acid, creatinine, amino acids and sugars. These substances may he determined on the basis of their colour effects, not only qualitatively but also quantitatively. For the quantitative determination of urea in aqueous solutions, a quinoline ligand can be synthesised which can be used not only for urea but also for thiourea ~see Tetrahedron Letters, 3, 309-312/1978).
The process of the invention with the use of the novel dyestuff systems is of great interest since it readily permits a plenitude of measurements and calculations which extend, for example, to the ion selectivity, caused by the complex constants with regard to various ions and the influence of ions of particular charge density on the chromo-phore system of the most va~ied types of dyestuffs with regard to extinction ànd absorption properties. In many cases, pre-liminary calculations with molecule orbi~al methods arepossible.
Another structural variant of the intramolecular ionophore/chromophore combination of the (XV) or (XVI) type is shown in another manner influencing selectivity: the cations fixed in the crown ether hollow space are additionally co-ordinated by the donor centre present on a comparatively short or comparatively long side chain, which can be a phenolic or CH20 ~ group or C00 ~3 or S03 ~3. Two influences are here combined: the negative charge of the side chain bonds the cation electrostatically more strongly than would be possible by a crown ether ring because of the ion-dipole reciprocal action. However, the cation must, nevertheless, pass into the crown ether ring, i.e., it is there tested and selected for itssize. Overall, higher complex constants are experimentally observed on the basis of the stronger bonding of the cation to the ligands, a higher selectivity of the cation embedded in the crown ether ring, as well as, as a result thereoE, a strong influencing of the chromophore system because of the favourable position of the equilibrium in comparison with the previously described ligands and a special selectivity towards cations with high charge density, for example, calcium ions. Such ligand systems, after optimisation to certain cations, can possibly be used for the speci-Eic differentiation of simple/plural charged cations:-H3C ~` ~ (XV) -m ~C~ - '~
2 ~ ~ N ~ (XVI) [CH2 ~COO (~7 .
~ - 24 -H3C 0 /~\ A ~ CH3 E3C ~ ~ ~ r-~ ~ ~ N ~ N
CH -~ ~ 2 The process according to the present invention provides a series of novel and practical uses:
a) With the help of the crown ether dyestuffs, indicators and probes can be developed, for example, for the detection of the phase trans~er of salts, for the study of ion trans-port by lipophilic media, for example, synthetic and biological membranes, i.e., for investigations in connection with membranological research from a physiological or pathological 0 ~point of view.
b) Another use is the possibility of ion-selectively colouring tissue sections. A possibly quantitative evaluation hereby permits conclusions regarding the nature and concent-ration of ions which are made visible by the displacement of AmaX and extinction.
c) The use of chromophore cryptand systems ensures, especially in aqueous media, not only a substantially higher complex stability but also an improved selectivity. Besides the qualitative and quantitative photometric detection o-f the alkali metals and alkaline earth metals, as well as of the ammonium ions, with the help of host chromophores of various hollow space size in solutions, for example, in blood or sera and other body fluids, these can also be identified on carrier materials. The use as spray reagents for ion chromatograms or silicone prints in the field of medical diagnosis is hereby possible. In the manner of thermography, either by spraying areas or tissue or their imprints with ion-selective dye-stuffs and fluorescent dyestuffs, physiological or pathological salt concentrations can be made visible. From this there follows the possibility of differentiating disease.
d) From the knowledge of the complexing of organic ammonium and guanidinium salts, as well as of neutral mole-cules, for example, urea and CH-acid compounds, it is, according to the present invention, now possible to carry out detection with appropriate host chromophores. The useful-ness from a medicinal or biochemical point o~ view hereby lies in the specific photometric detection of certain sub-stances which are peculiar to the body and the kinetic monitoring of enzymatic processes, Having regard thereto, there can be considered the attachment of lipophilic hollow spaces in the form of endolipiphilic or endohydrophobic macrocyclic compounds with exohydrophilic or exopolarophilic molecular peripheries on sensitive i.e~ chromophore-influencing, positions of one of the dyestuffs which permit the enveloping of an appropriate neutral particle and, probably due to hydrogen bridges, bring about a change of the absorption or extinction.
The invention is further illustrated by reference to the accompanying drawings, referred to more particularly in the examples in which:
FIGURE 1 shows graphically the absorption spectra of 2,4-dinitro-4'-(4,7,10,13-tetraoxa-1-azocyclopentadec-l-yl)-stilbene with various metal ions, according to Example 3, FIGURE 2 shows graphically the absorption spectra of 2,4-dinitro-4'-(4,7,10,13,16-pentaoxa-1-azacyclooctadec-l-yl)-stilbene and its com-plexes with ammonium ions and various metal ions, according to Example 6, FIGURE 3 illustrates graphically the absorption spectra of a blue dyestuff and its com-plexes with various metal ions, according to Example 7, FIGURE 4 illustrates graphically the absorption spectra of a dyestuff and its complexes with various metal ions, according to Example 8, FIGURE S illustrates graphically the absorption spectra of 1,8-bis[l-(4-dimethylamino-phenylimino)-~-benzoquin-3-oxy~-3,6-dioxaoctane and its complexes with various metal ions, according to Example 9, and FIGURE 6 illustrates graphically the absorption spectra of a dyestuff and its complexes .-with various metal ions, according to .
. Example 10.
: The following Examples are given for the purpose of ~:~ 20 illustrating the present invention:
: Example 1 .
Photometric determination of potassium with an ion-selective dyestuff which contains the chromophore covalently bound via a mesomeric system An ion-selective dye.stuff of general formula (IV')`
(n = 1) is dissolved in chloroform and shaken up with a solution which contains potassium ions. The dyestuff com-plexed with potassium passes over into the aqueous phase and can be determined quantitatively at 600nm in a photometer, depending upon the activity of the potassium ions present.
i~
The dyestuff (5-nitro-1,3-thiazole-2-azo-[3-(12- -hydroxy-1,4,7,10-tetraoxadodecyl]-4-hydroxybenzene) can be prepared by reacting 2-amino~5-nitrothiazole in 85% phosphoric acid with sodium nitrite and subsequently adding benzo[l5] crown-
5. ~he intensively red suspension is mixed with water and extracted with chLoroform. After drying with anhydrous sodium sulphate and removing the solvent in a vacuum, it is separated by column chromatography on silica gel. With the use of chloroform, there is first obtained the ring-closed dyestuff 5-nitro-1,3-thiazole-2-azo[1,4,7,10,13-pentaoxa[13]-(3,4)-benzophane (VII). By the addition of 5% ethanol to the eluent, there is obtained the ring-open dyestuff (IV') (n = 1).
Both dyestuffs can be obtained in crystalline form by dis-solving in ethyl acetate and adding petroleum ether (b. p.
60 - 90C.), The melting points are 169 - 171C. (VII) and 112 - 115C. (IV').
Example 2 For the investigation of the properties of the ion-selective dyestuffs mentioned in Example 1 for test strips, a solution of the dyestuff is applied dropwise to a strip of filter paper and the strip then dried. The paper strip treated in this manner becomes deep red coloured upon the application thereto of sample solutions which contain potassium ions.
Besides this qualitative method of detection, the coloured strip can also be quantitatively evaluated in a reflection photometer.
Exal~le 3 Selective photometric determination of potassium with a ligand which carries a covalently-bound chromophore.
A compound of general formula (IV), in which n is 1 and Rl is the radical:-L
02N ~ CH=CH-~2 is obtained by reacting 2,4-dinitrotoluene for 3 hours with ~-(~-formylphenyl)-aza[15] crown-5 and a few drops of piperidine at lOO~C. The solidified melt is dissolved in a little ethyl acetate, filtered and the filtrate evaporated in a vacuum.
The residue is purified by column chromatography on silica gel, using ethyl acetate/5-1~/O ethanol. Upon concentrating the eluate, 2,4-dinitro-~'-t4,7,10,13-tetraoxa-1-azocyclo-pentadec-l-yl)-stilbene crystallises out; m~p. 143 - 146C.
7.5 mg. Dyestuff are dissolved in 250 ml. methanol.
Furthermore, O.lM solutions are prepared of calcium chloride, sodium chloride, potassium chloride, lithium chloride and magnesium chloride, in each case in O.lM triethanolamine hydrochloride/sodium hydroxide buffer (pH 7.0).
There is also prepared a series of solutions with 0.001 to O.lM potassium chloride, again in each case in O.lM
triethanolamine hydrochloride/sodium hydroxide buffer (pH 7.0).
3ml. Amounts of dyestuff solution are mixed with 0.5 ml. of the particular salt solution to be investigated and the absorption behaviour determined in a photometer at 366 nm. This dyestuff shows a spectrum towards potassium which is different from that in the presence of lithium, sodium, calcium, barium and magnesium. The concentration of the potassium ions is directly proportional to the extinction.
The absorption spectra of some of these metal ions are given in Figure 1 of the accompanying drawings.
_ 29 -Example 4 Photometric determination of calcium and lithium with a heterocyclic crown ether of formula (IV) in which n is 1 and Rl is 0 ~ N- and thus the chromophore is covalently attached.
1,4-Benzoquinone-(4,7,10,13-tetraoxa~l-azacyclo-pentadec-l-yl)-phenylimine is obtained by the oxidative coupling of N-(4-aminophenyl)-aza[15] crown-5 with phenol.
Furthermore, silver nitrate is reacted with sodium chloride and some starch, sodium carbonate and phenol are added there-to and a solution of N-(4-aminophenyl)aza[15]-crown-5 in con-centrated hydrochloric acid added thereto dropwise. After the reaction, the reaction mixture is stirred with ethyl acetate, dried with anhydrous sodium sulphate and evaporated in a vacuum. The residue crystallises from tetrahydrofuran with diethyl ether at 0C.; m.p. 55 - 56C.; ~max = 583, log ~ =
4.77.
The dyestuff is dissolved in chloroform (E578 0.8) and mixed with an equal amount by volume of an a~ueous salt solution. With lithium and calcium, a colour change was measured at 578 nm which is directly proportional to the ions to be determined.
By the oxidative coupling of ~-naphthol with N-(4-aminophenyl)-aza[15]-crown-5, there is obtained 1,4-naphth-quinone-4-(4,7,10,13-tetr~oxa-1-azacyclopentadec-1-yl)-phenyl-amine; m.p. 124 - 125C.; ~max = 577~ log ~ = 4.41. The colour displacement by metal ions corresponds to that of the benzoquinone derivative.
.
4~
Exam~le 5 Orange-coloured 2,4-dinitrophenylhydrazinium hydrochlorlde (~max 395 nm) is dissolved in methanol/water and mixed with molar amounts of crown ether [18] crown-6 A change of the absorption (brightening) is observed between the free dyestuff and the dyestuff bound in the ionophore compleY~ (~maY ~384 nm). To this is added an aqueous methanolic solution containing potassium or sodium ions, for exam~le, a solution of sodium perchlorate or potassium thiocyanate, a deepening of the colour being observed since the liberated dyestuff again displays the initial absorption maximum.
If the crown ether in the above Example is replaced by another ionophore, such as valinomycin or nonactin, then, depending upon the ionophore, more or less strongly marked eolour effects or absorption changes are observed.
Example 6 In a manner analogous to that: described in Example 3, 2,4-dinitrotoluene is heated with N-(E~formylphenyl)-aza[18]-erown-6 and a few drops of piperidine for 4 to 5 hours at 100 - 110C. The reaction mixture is dissolved in dichloro-methane and ehromatographed on silica gel with ethyl aeetate/
ehloroform (1:1 v/v). After recrystallisation from ethyl aeetate/ethanol (1:1 v/v), there is obtained the dyestuff 2,4-dinitro-4'-(4,7,10,13,16-pentaoxa-1-aza-eyclooctadec-1-yl)-stilbene; m.p. 90 - 91C. The absorption spectra of the dye-stuff and of its complexes w th barium, ammonium, calcium, potassium, sodium, rubidium and lithium ions are given in Figure 2 of the aceompanying drawings.
Example 7 In a manner analogous to that described in Example ~, by the oxidative coupling of N-(2-hydroxybenzyl)-monoaza[15]~
crown-5 with 4-a~ no-N,N-dimethylaniline, there is obtained a blue dyestuff which can be extracted with ethyl acetate.
After drying the extract with anhydrous magnesium sulphate, filtering and evaporating the filtrate in a vacuum, the residue obtained is chromatographed on silica gel with ethanol/ethyl acetate (1:9 v/v). The absorption spectra of the dyestuff and of its complexes with sodium, potassium, rubidium, magnesium, calcium, barium, lithium and nickel ions are given in Figure 3 of the accompanying drawings.
Example 8 In a manner analogous to that described in Example 7, from N-(2-hydroxybenzyl)-monoaza~18]crown-6, there is obtained a dyestuff, the absorption spectra of the free and complexed state of which are given in Figure 4 of the accompanying drawings.
Example 9 The spectra of 1,8-bis-[1-(4-dimethylaminophenylimino)-~-benzoquin-3-oxy]-3,6-dioxaoctane in the free form and in the complexed forms with potassium, sodium, lithium, rubidium, barium and calcium ions are given in Figure 5 of the accompany-ing drawings. This compound can be prepared by reducing E~
nitroso-N-N-dimethylaniline with hydrochloric acid and zinc dust and reacting with 1,8-bis-(2-hydroxyphenoxy)-3,8-dioxa-octane in the presence of sodium hydroxide and potassium dichromate. Upon acidification with glacial acetic acid, the blue dyestuf precipitates out. After drying out with acetone, it is cooled, filtered off with suction and washed with a little acetone. It is then extracted with acetone in a hot extractor and chromatographed on silica gel with ethyl acetate.
The dyestuff is obtained by adding 5% ethanol to the ethyl acetate solution. After crystallisation from acetone/r-heptane, the dyestuff melts a-t 112 - 115C.; ~ n x = 572 log ~ =
~.73.
Exam~le 10 The spectrum of the dyestuff (IV), in which Rl is ~ and n is 1, as well as the spectra of the metal ion complexes with calcium, barium, sodium, lithium and potassium, are given in Figure 6 of the accompanying drawings. This dyestuff is obtained by suspending N-(4-aminophenyl)-aza[15]-crown-5 and 1,4-naphthoquinone with copper acetate monohydrate in ethanol, boiling for 1 hour and passing through air. After removing the solvent in a vacuum, the residue is mixed with water, extracted with di-chloromethane, dried with anhydrous magnesium sulphate and evaporated in a vacuum. Tha oily residue is purified on silica gel with dichloromethane. The eluate initially contains excess naphthoquinone and, after the addition of 5% ethanol, contains the desired dyestuff, which can be recrystallised from ethyl acetate.
Both dyestuffs can be obtained in crystalline form by dis-solving in ethyl acetate and adding petroleum ether (b. p.
60 - 90C.), The melting points are 169 - 171C. (VII) and 112 - 115C. (IV').
Example 2 For the investigation of the properties of the ion-selective dyestuffs mentioned in Example 1 for test strips, a solution of the dyestuff is applied dropwise to a strip of filter paper and the strip then dried. The paper strip treated in this manner becomes deep red coloured upon the application thereto of sample solutions which contain potassium ions.
Besides this qualitative method of detection, the coloured strip can also be quantitatively evaluated in a reflection photometer.
Exal~le 3 Selective photometric determination of potassium with a ligand which carries a covalently-bound chromophore.
A compound of general formula (IV), in which n is 1 and Rl is the radical:-L
02N ~ CH=CH-~2 is obtained by reacting 2,4-dinitrotoluene for 3 hours with ~-(~-formylphenyl)-aza[15] crown-5 and a few drops of piperidine at lOO~C. The solidified melt is dissolved in a little ethyl acetate, filtered and the filtrate evaporated in a vacuum.
The residue is purified by column chromatography on silica gel, using ethyl acetate/5-1~/O ethanol. Upon concentrating the eluate, 2,4-dinitro-~'-t4,7,10,13-tetraoxa-1-azocyclo-pentadec-l-yl)-stilbene crystallises out; m~p. 143 - 146C.
7.5 mg. Dyestuff are dissolved in 250 ml. methanol.
Furthermore, O.lM solutions are prepared of calcium chloride, sodium chloride, potassium chloride, lithium chloride and magnesium chloride, in each case in O.lM triethanolamine hydrochloride/sodium hydroxide buffer (pH 7.0).
There is also prepared a series of solutions with 0.001 to O.lM potassium chloride, again in each case in O.lM
triethanolamine hydrochloride/sodium hydroxide buffer (pH 7.0).
3ml. Amounts of dyestuff solution are mixed with 0.5 ml. of the particular salt solution to be investigated and the absorption behaviour determined in a photometer at 366 nm. This dyestuff shows a spectrum towards potassium which is different from that in the presence of lithium, sodium, calcium, barium and magnesium. The concentration of the potassium ions is directly proportional to the extinction.
The absorption spectra of some of these metal ions are given in Figure 1 of the accompanying drawings.
_ 29 -Example 4 Photometric determination of calcium and lithium with a heterocyclic crown ether of formula (IV) in which n is 1 and Rl is 0 ~ N- and thus the chromophore is covalently attached.
1,4-Benzoquinone-(4,7,10,13-tetraoxa~l-azacyclo-pentadec-l-yl)-phenylimine is obtained by the oxidative coupling of N-(4-aminophenyl)-aza[15] crown-5 with phenol.
Furthermore, silver nitrate is reacted with sodium chloride and some starch, sodium carbonate and phenol are added there-to and a solution of N-(4-aminophenyl)aza[15]-crown-5 in con-centrated hydrochloric acid added thereto dropwise. After the reaction, the reaction mixture is stirred with ethyl acetate, dried with anhydrous sodium sulphate and evaporated in a vacuum. The residue crystallises from tetrahydrofuran with diethyl ether at 0C.; m.p. 55 - 56C.; ~max = 583, log ~ =
4.77.
The dyestuff is dissolved in chloroform (E578 0.8) and mixed with an equal amount by volume of an a~ueous salt solution. With lithium and calcium, a colour change was measured at 578 nm which is directly proportional to the ions to be determined.
By the oxidative coupling of ~-naphthol with N-(4-aminophenyl)-aza[15]-crown-5, there is obtained 1,4-naphth-quinone-4-(4,7,10,13-tetr~oxa-1-azacyclopentadec-1-yl)-phenyl-amine; m.p. 124 - 125C.; ~max = 577~ log ~ = 4.41. The colour displacement by metal ions corresponds to that of the benzoquinone derivative.
.
4~
Exam~le 5 Orange-coloured 2,4-dinitrophenylhydrazinium hydrochlorlde (~max 395 nm) is dissolved in methanol/water and mixed with molar amounts of crown ether [18] crown-6 A change of the absorption (brightening) is observed between the free dyestuff and the dyestuff bound in the ionophore compleY~ (~maY ~384 nm). To this is added an aqueous methanolic solution containing potassium or sodium ions, for exam~le, a solution of sodium perchlorate or potassium thiocyanate, a deepening of the colour being observed since the liberated dyestuff again displays the initial absorption maximum.
If the crown ether in the above Example is replaced by another ionophore, such as valinomycin or nonactin, then, depending upon the ionophore, more or less strongly marked eolour effects or absorption changes are observed.
Example 6 In a manner analogous to that: described in Example 3, 2,4-dinitrotoluene is heated with N-(E~formylphenyl)-aza[18]-erown-6 and a few drops of piperidine for 4 to 5 hours at 100 - 110C. The reaction mixture is dissolved in dichloro-methane and ehromatographed on silica gel with ethyl aeetate/
ehloroform (1:1 v/v). After recrystallisation from ethyl aeetate/ethanol (1:1 v/v), there is obtained the dyestuff 2,4-dinitro-4'-(4,7,10,13,16-pentaoxa-1-aza-eyclooctadec-1-yl)-stilbene; m.p. 90 - 91C. The absorption spectra of the dye-stuff and of its complexes w th barium, ammonium, calcium, potassium, sodium, rubidium and lithium ions are given in Figure 2 of the aceompanying drawings.
Example 7 In a manner analogous to that described in Example ~, by the oxidative coupling of N-(2-hydroxybenzyl)-monoaza[15]~
crown-5 with 4-a~ no-N,N-dimethylaniline, there is obtained a blue dyestuff which can be extracted with ethyl acetate.
After drying the extract with anhydrous magnesium sulphate, filtering and evaporating the filtrate in a vacuum, the residue obtained is chromatographed on silica gel with ethanol/ethyl acetate (1:9 v/v). The absorption spectra of the dyestuff and of its complexes with sodium, potassium, rubidium, magnesium, calcium, barium, lithium and nickel ions are given in Figure 3 of the accompanying drawings.
Example 8 In a manner analogous to that described in Example 7, from N-(2-hydroxybenzyl)-monoaza~18]crown-6, there is obtained a dyestuff, the absorption spectra of the free and complexed state of which are given in Figure 4 of the accompanying drawings.
Example 9 The spectra of 1,8-bis-[1-(4-dimethylaminophenylimino)-~-benzoquin-3-oxy]-3,6-dioxaoctane in the free form and in the complexed forms with potassium, sodium, lithium, rubidium, barium and calcium ions are given in Figure 5 of the accompany-ing drawings. This compound can be prepared by reducing E~
nitroso-N-N-dimethylaniline with hydrochloric acid and zinc dust and reacting with 1,8-bis-(2-hydroxyphenoxy)-3,8-dioxa-octane in the presence of sodium hydroxide and potassium dichromate. Upon acidification with glacial acetic acid, the blue dyestuf precipitates out. After drying out with acetone, it is cooled, filtered off with suction and washed with a little acetone. It is then extracted with acetone in a hot extractor and chromatographed on silica gel with ethyl acetate.
The dyestuff is obtained by adding 5% ethanol to the ethyl acetate solution. After crystallisation from acetone/r-heptane, the dyestuff melts a-t 112 - 115C.; ~ n x = 572 log ~ =
~.73.
Exam~le 10 The spectrum of the dyestuff (IV), in which Rl is ~ and n is 1, as well as the spectra of the metal ion complexes with calcium, barium, sodium, lithium and potassium, are given in Figure 6 of the accompanying drawings. This dyestuff is obtained by suspending N-(4-aminophenyl)-aza[15]-crown-5 and 1,4-naphthoquinone with copper acetate monohydrate in ethanol, boiling for 1 hour and passing through air. After removing the solvent in a vacuum, the residue is mixed with water, extracted with di-chloromethane, dried with anhydrous magnesium sulphate and evaporated in a vacuum. Tha oily residue is purified on silica gel with dichloromethane. The eluate initially contains excess naphthoquinone and, after the addition of 5% ethanol, contains the desired dyestuff, which can be recrystallised from ethyl acetate.
Claims (35)
1. A process for the determination of an entity selected from the group consisting of ions, polar substances and lipophilic substances in liquids, comprising allowing the entity to be determined to act upon a complex ligand or host molecule which is selective towards the entity to be determined, said complex ligand or host molecule being associated with at least one chromophore and, thereafter, measuring the change of extinction or wavelength displace-ment of the chromophore.
2. A process according to claim 1, wherein said entity is allowed to act upon a complex ligand.
3. A process according to claim 1, wherein said entity is allowed to act upon a host molecule.
4. A process according to claim 1, wherein said complex ligand ox host molecule is attached by a direct covalent bond, heteropolar bond, hydrogen bridge or hydro-phobic bond to said at least one chromophore.
5. A process according to claim 1, wherein said complex ligand or host molecule contains the chromophore in the form of an inclusion complex.
6. A process according to claim 1, wherein the selective complex ligand or the selective host molecule is a cyclic or acyclic compound which, with regard to the entity to be determined is already present as a complex former or, in the presence thereof, assumes the structure necessary for complex formation or adduct formation.
7. A process according to claim 1 or 6, wherein the selective complex ligand or the selective host molecule is a cyclic or open-chained oligoether, polyether, oligoester, polyester, oligoamide or polyamide or a pure hydrocarbon structure, or comprises a combination of these functions.
8. A process according to claim 2, wherein the selective complex ligand is a crown ether, cryptand, podand or derivative thereof.
9. A process according to claim 8, wherein the ligand is said derivative and has bridges or chains which contain oligo or polyethylene glycol groupings or other hetero atom-containing groupings.
10. A process according to claim 2, wherein the complex ligand is a cyclic peptide or a peptide which, in the presence of an ion or polar substance to be determined, assumes a secondary, tertiary or quaternary structure necessary for complex formation, or is a tetrahydrofuran-containing ester-linked macrolide.
11. A process according to claim 10, wherein the complex ligand is valinomycin, gramacidin, nonactin or a related ionophore compound.
12. A process according to claim 1, wherein said complex ligand or host molecule is covalently-, heteropolar- or hydrophobically-bound to said chromophore, and said chromophore is a dyestuff or a fluorescent dyestuff or a chromogen, the absorption spectrum of which is changed by reciprocal action with a guest particle, by charge displacement or disturbance of the mesomeric system in at least one of the base and excited states.
13. A process according to claim 12, wherein the chromo-phore has a polyene, meriquinoid, quinone, azo, pyrrole, merocyanin, indigo, indophenol, stilbene, azomethine, anthraquinone, naphthoquinone, cyanine, phthalein, poly-methine or alizarine structure.
14. A process according to claim 5, wherein the chromophore contained in the inclusion complex is an acidic dyestuff or a salt thereof.
15. A process according to claim 14, wherein the chromo-phore is a lithium, sodium, potassium, ammonium, calcium, alkylammonium or magnesium salt of the acidic dyestuff.
16. A process according to claim 14 or 15, wherein the dyestuff contains a carboxylate, sulphonate, phenolate or thiophenolate grouping.
17. A process according to claim 11, 12 or 14, wherein the entity to be detected is an anion.
18. A process according to claim 1, 12 or 14, wherein the entity to be detected is a cation.
19. A process according to claim 1, 12 or 14, wherein the entity to be detected is an ammonium, alkali metal, alkaline earth metal or a heavy metal ion.
20. A process according to claim 1, 12 or 14, wherein the entity to be detected is a lithium, sodium, potassium, magnesium, calcium, iron, zinc, copper, cobalt, nickel, molybdenum or chromium ion.
21. A process according to claim 1, 12 or 14, wherein the entity to be detected is an anion selected from the group consisting of chloride, bromide, iodide, sulphate, nitrate, nitrite, phosphate, diphosphate, triphosphate, hydrogen phosphate and hydrogen carbonate ion.
22. A process according to claim 1, 12 or 14, wherein the entity to be detected is urea, thiourea, guanidine, uric acid, choline, creatinine, an amino acid or a sugar.
23. A process according to claim 1, 12 or 14, wherein the entity to be detected is a lipophilic guest molecule, or a lipid.
24. A process according to claim 1, 12 or 14, wherein the entity to be detected is a steroid.
25. A process according to claim 1, 12 or 14, wherein the entity to be detected is cholesterol, a triglyceride or lecithin.
26. A process according to claim 1, wherein said chromophore is linked with several identical or different complex ligands or host molecules.
27. A process according to claim 1, wherein a complex ligand or host molecule is linked with several identical or different chromophores.
28. A process according to claim 1, wherein the complex ligand or host molecule is applied to or incorporated into a solid carrier, synthetic resin film, glass, aluminium oxide, silicone oxide, natural or synthetic fibre or metal.
29. A reagent for the determination of an entity selected from the group consisting ofions, polar substances and lipophilic substances in liquids, said reagent containing a complex ligand or host molecule which is selective towards the entity to be determined, said complex ligand or host molecule being associated with at least one chromophore
30. A reagent according to claim 29, wherein said complex ligand or host molecule is attached by a direct covalent bond, heteropolar bond, hydrogen bridge or hydro-phobic bond to said at least one chromophore.
31. A reagent according to claim 29, wherein said complex ligand or host molecule contains the chromophore in the form of an inclusion complex.
32. A test device comprising an inert substrate having applied thereto a reagent as defined in claim 29.
33. A test device according to claim 32, wherein said substrate is paper or a synthetic resin film.
34, A test device according to claim 32, wherein said substrate is glass, aluminium oxide, silicon oxide or a metal.
35. A test device according to claim 32, wherein said substrate comprises natural or synthetic fibres or textile material.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19782842862 DE2842862A1 (en) | 1978-10-02 | 1978-10-02 | METHOD FOR DETERMINING ION, POLAR AND / OR LIPOPHILE SUBSTANCES IN LIQUIDS |
| DEP2842862.8 | 1978-10-02 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1142841A true CA1142841A (en) | 1983-03-15 |
Family
ID=6051114
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000336828A Expired CA1142841A (en) | 1978-10-02 | 1979-10-02 | Process and a reagent for the determination of ions and of polar and/or lipophilic substances in fluids |
Country Status (9)
| Country | Link |
|---|---|
| US (1) | US4367072A (en) |
| EP (1) | EP0010615B1 (en) |
| JP (1) | JPS5548643A (en) |
| AT (1) | ATE8713T1 (en) |
| AU (1) | AU515023B2 (en) |
| CA (1) | CA1142841A (en) |
| DE (2) | DE2842862A1 (en) |
| SU (1) | SU1135436A3 (en) |
| ZA (1) | ZA794814B (en) |
Families Citing this family (112)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4734375A (en) * | 1980-05-27 | 1988-03-29 | Miles Laboratories, Inc. | Ion specific test method |
| CA1161346A (en) * | 1980-05-27 | 1984-01-31 | Steven C. Charlton | Ion specific analytical element |
| JPS57194787A (en) * | 1981-05-28 | 1982-11-30 | Ajinomoto Co Inc | Culture medium for animal cell |
| DE3202779A1 (en) * | 1982-01-28 | 1983-08-04 | Merck Patent Gmbh, 6100 Darmstadt | POTASSIUM REAGENT AND METHOD FOR DETERMINING POTASSIUM IONS |
| JPS59145022A (en) * | 1983-02-08 | 1984-08-20 | Japan Atom Energy Res Inst | Separation of lithium isotope |
| US4649123A (en) * | 1983-05-12 | 1987-03-10 | Miles Laboratories, Inc. | Ion test means having a hydrophilic carrier matrix |
| CA1222438A (en) * | 1983-05-12 | 1987-06-02 | Steven C. Charlton | Unified test means for ion determination |
| CA1226792A (en) * | 1983-05-12 | 1987-09-15 | Steven C. Charlton | Ion test means having a hydrophilic carrier matrix |
| CA1225574A (en) * | 1983-11-07 | 1987-08-18 | Anand Kumar | Reflective particle-containing solvent extraction reagent composition |
| US4670218A (en) * | 1984-02-24 | 1987-06-02 | Miles Laboratories, Inc. | Ion test means having a porous carrier matrix |
| CA1253057A (en) * | 1984-02-24 | 1989-04-25 | Mary L. Gantzer | Ion test means having a porous carrier matrix |
| DE3424065A1 (en) * | 1984-06-29 | 1986-01-09 | Bayer Ag, 5090 Leverkusen | METHOD FOR ACTIVATING SUBSTRATE SURFACES FOR ELECTRIC METALLIZATION |
| CA1260387A (en) * | 1984-07-17 | 1989-09-26 | Technicon Instruments Corporation | Polymeric single layer analytical element |
| DE3436971A1 (en) * | 1984-10-09 | 1986-06-12 | Bayer Ag, 5090 Leverkusen | SEMI-FINISHED PRODUCTION BOARDS |
| US4734376A (en) * | 1985-02-04 | 1988-03-29 | The President & Trustees Of The Miami University | Process for the spectrophotometric measurement of lithium ion employing a chromogenic aza-12-crown-4- ether |
| JPS62161053A (en) * | 1986-01-10 | 1987-07-17 | Konishiroku Photo Ind Co Ltd | Analytical element |
| US4820647A (en) * | 1986-12-03 | 1989-04-11 | Biotrack, Inc. | Method for detecting a metal ion in an aqueous environment |
| US4795712A (en) * | 1987-04-10 | 1989-01-03 | Eastman Kodak Company | Calcium complexing dyes and their use in analytical compositions, elements and methods |
| US5384247A (en) * | 1987-04-10 | 1995-01-24 | Boehringer Mannheim, Gmbh | Determination of sodium ions in fluids |
| US5073501A (en) * | 1987-04-15 | 1991-12-17 | Technicon Instruments Corporation | Method for determining the concentration of cations in a sample |
| US4994395A (en) * | 1987-04-15 | 1991-02-19 | Technicon Instruments Corporation | Chromogenic cryptand reagents and methods for determining cation concentrations in test samples containing interfering cations |
| US5096831A (en) * | 1987-04-15 | 1992-03-17 | Miles Inc. | Reagents for determining cations |
| US5177221A (en) * | 1987-04-15 | 1993-01-05 | Miles Inc. | Chromogenic hemispherands and their preparation |
| US5045475A (en) * | 1987-04-15 | 1991-09-03 | Technicon Instruments Corporation | Reagent for determining cations |
| US4992381A (en) * | 1987-04-15 | 1991-02-12 | Technicon Instruments Corporation | Method of detecting electrolytes in an aqueous test sample using chromogenic hemispherands and test device therefor |
| US5145787A (en) * | 1987-04-15 | 1992-09-08 | Miles, Inc. | Method for determining the concentration of cations in a sample |
| US4808539A (en) * | 1987-04-15 | 1989-02-28 | Technicon Instruments Corporation | Compounds, reagents and procedures for determining cations |
| US5001048A (en) * | 1987-06-05 | 1991-03-19 | Aurthur D. Little, Inc. | Electrical biosensor containing a biological receptor immobilized and stabilized in a protein film |
| US5192507A (en) * | 1987-06-05 | 1993-03-09 | Arthur D. Little, Inc. | Receptor-based biosensors |
| US5162525A (en) * | 1987-07-31 | 1992-11-10 | Allied-Signal Inc. | Fluorogenic and chromogenic three-dimensional ionophores as selective reagents for detecting ions in biological fluids |
| US5049673A (en) * | 1987-10-30 | 1991-09-17 | The Regents Of The University Of California | Fluorescent indicator dyes for calcium working at long wavelengths |
| DE58907029D1 (en) * | 1988-08-26 | 1994-03-31 | Moeller Willi Ag | Test device for the optical determination of cations, anions or electrically neutral ionogenic species and test method using this test device. |
| CA2002917C (en) * | 1988-11-14 | 1998-01-06 | Akwasi Minta | Fluorescent indicator dyes for alkali metal cations |
| US5232858A (en) * | 1989-04-25 | 1993-08-03 | Avl Medical Instruments Ag | Method for quantitative determination of at least one chemical parameter of a sample medium |
| WO1991005605A1 (en) * | 1989-10-10 | 1991-05-02 | Kosak Kenneth M | Cyclodextrin labels for immunoassay and biochemical analysis |
| DE4012750A1 (en) * | 1990-04-21 | 1991-10-24 | Hoechst Ag | FERROELECTRIC LIQUID CRYSTAL DISPLAY WITH HIGH CONTRAST AND HIGH BRIGHTNESS |
| DE69123519T2 (en) * | 1990-08-06 | 1997-04-03 | Bayer Ag | Method and device for determining ions |
| US5154890A (en) * | 1990-11-07 | 1992-10-13 | Hewlett-Packard Company | Fiber optic potassium ion sensor |
| US5132436A (en) * | 1991-05-28 | 1992-07-21 | Eastman Kodak Company | Intermediates for making 14-crown-4-ether derivatives |
| US5245021A (en) * | 1991-05-28 | 1993-09-14 | Eastman Kodak Company | 14-crown-4-ether derivatives |
| US5705463A (en) * | 1993-02-24 | 1998-01-06 | Tech Spray, Inc. | Composition and process for removal of ionic salt deposits |
| US5604191A (en) * | 1993-02-24 | 1997-02-18 | Tech Spray, Inc. | Composition for removal of ionic salt deposits |
| US5643356A (en) | 1993-08-05 | 1997-07-01 | Kimberly-Clark Corporation | Ink for ink jet printers |
| US5865471A (en) | 1993-08-05 | 1999-02-02 | Kimberly-Clark Worldwide, Inc. | Photo-erasable data processing forms |
| US5733693A (en) | 1993-08-05 | 1998-03-31 | Kimberly-Clark Worldwide, Inc. | Method for improving the readability of data processing forms |
| US5721287A (en) | 1993-08-05 | 1998-02-24 | Kimberly-Clark Worldwide, Inc. | Method of mutating a colorant by irradiation |
| US6017471A (en) | 1993-08-05 | 2000-01-25 | Kimberly-Clark Worldwide, Inc. | Colorants and colorant modifiers |
| US5773182A (en) | 1993-08-05 | 1998-06-30 | Kimberly-Clark Worldwide, Inc. | Method of light stabilizing a colorant |
| CA2120838A1 (en) | 1993-08-05 | 1995-02-06 | Ronald Sinclair Nohr | Solid colored composition mutable by ultraviolet radiation |
| US5681380A (en) | 1995-06-05 | 1997-10-28 | Kimberly-Clark Worldwide, Inc. | Ink for ink jet printers |
| US6017661A (en) | 1994-11-09 | 2000-01-25 | Kimberly-Clark Corporation | Temporary marking using photoerasable colorants |
| US6211383B1 (en) | 1993-08-05 | 2001-04-03 | Kimberly-Clark Worldwide, Inc. | Nohr-McDonald elimination reaction |
| US5645964A (en) | 1993-08-05 | 1997-07-08 | Kimberly-Clark Corporation | Digital information recording media and method of using same |
| US6033465A (en) | 1995-06-28 | 2000-03-07 | Kimberly-Clark Worldwide, Inc. | Colorants and colorant modifiers |
| AT401823B (en) * | 1993-09-30 | 1996-12-27 | Avl Verbrennungskraft Messtech | OPTICAL INDICATOR FOR DETERMINING THE ACTIVITY OF AN ION IN A SAMPLE |
| US5418058A (en) * | 1993-10-04 | 1995-05-23 | The Regents Of The University Of California | Chemical microsensors |
| US7047171B1 (en) | 1995-12-08 | 2006-05-16 | Sproch Norman K | Method for the characterization of the three-dimensional structure of proteins employing mass spectrometric analysis and computational feedback modeling |
| US20060277017A1 (en) * | 1993-11-04 | 2006-12-07 | Sproch Norman K | Method for the characterization of the three-dimensional structure of proteins employing mass spectrometric analysis and computational feedback modeling |
| US5504327A (en) * | 1993-11-04 | 1996-04-02 | Hv Ops, Inc. (H-Nu) | Electrospray ionization source and method for mass spectrometric analysis |
| JP3393701B2 (en) * | 1994-03-18 | 2003-04-07 | 株式会社東北テクノアーチ | Spot test method for ionic samples |
| US6071979A (en) | 1994-06-30 | 2000-06-06 | Kimberly-Clark Worldwide, Inc. | Photoreactor composition method of generating a reactive species and applications therefor |
| US6242057B1 (en) | 1994-06-30 | 2001-06-05 | Kimberly-Clark Worldwide, Inc. | Photoreactor composition and applications therefor |
| US5739175A (en) | 1995-06-05 | 1998-04-14 | Kimberly-Clark Worldwide, Inc. | Photoreactor composition containing an arylketoalkene wavelength-specific sensitizer |
| US5685754A (en) | 1994-06-30 | 1997-11-11 | Kimberly-Clark Corporation | Method of generating a reactive species and polymer coating applications therefor |
| US6008268A (en) | 1994-10-21 | 1999-12-28 | Kimberly-Clark Worldwide, Inc. | Photoreactor composition, method of generating a reactive species, and applications therefor |
| ATE195815T1 (en) | 1995-06-05 | 2000-09-15 | Kimberly Clark Co | DYE PRECURSORS AND COMPOSITIONS CONTAINING SAME |
| US5798015A (en) | 1995-06-05 | 1998-08-25 | Kimberly-Clark Worldwide, Inc. | Method of laminating a structure with adhesive containing a photoreactor composition |
| US5811199A (en) | 1995-06-05 | 1998-09-22 | Kimberly-Clark Worldwide, Inc. | Adhesive compositions containing a photoreactor composition |
| US5747550A (en) | 1995-06-05 | 1998-05-05 | Kimberly-Clark Worldwide, Inc. | Method of generating a reactive species and polymerizing an unsaturated polymerizable material |
| US5786132A (en) | 1995-06-05 | 1998-07-28 | Kimberly-Clark Corporation | Pre-dyes, mutable dye compositions, and methods of developing a color |
| US5849411A (en) | 1995-06-05 | 1998-12-15 | Kimberly-Clark Worldwide, Inc. | Polymer film, nonwoven web and fibers containing a photoreactor composition |
| MX9705708A (en) | 1995-11-28 | 1997-10-31 | Kimberly Clark Co | Improved colorant stabilizers. |
| US5782963A (en) | 1996-03-29 | 1998-07-21 | Kimberly-Clark Worldwide, Inc. | Colorant stabilizers |
| US5855655A (en) | 1996-03-29 | 1999-01-05 | Kimberly-Clark Worldwide, Inc. | Colorant stabilizers |
| US6099628A (en) | 1996-03-29 | 2000-08-08 | Kimberly-Clark Worldwide, Inc. | Colorant stabilizers |
| US5891229A (en) | 1996-03-29 | 1999-04-06 | Kimberly-Clark Worldwide, Inc. | Colorant stabilizers |
| EP0894261B1 (en) | 1996-04-16 | 2005-06-15 | Novartis AG | Covalently immobilized fluoroionophores as optical ion sensors |
| EP0907641B1 (en) | 1996-04-18 | 2002-06-12 | Novartis AG | Fluoroionophores and their use in optical ion sensors |
| JP4223555B2 (en) | 1996-07-22 | 2009-02-12 | ノバルティス アクチエンゲゼルシャフト | Covalently immobilized fluoroionophore for optical ion sensor |
| US5747345A (en) * | 1996-08-15 | 1998-05-05 | Johnson & Johnson Clinical Diagnostics, Inc. | Calcium specific diaza-18-crown-6-ether chromoionophores |
| US6245574B1 (en) * | 1997-07-03 | 2001-06-12 | Novartis Ag | Sensors |
| US6524379B2 (en) | 1997-08-15 | 2003-02-25 | Kimberly-Clark Worldwide, Inc. | Colorants, colorant stabilizers, ink compositions, and improved methods of making the same |
| JP2002517523A (en) | 1998-06-03 | 2002-06-18 | キンバリー クラーク ワールドワイド インコーポレイテッド | Novel photoinitiator and its use |
| JP2002517540A (en) | 1998-06-03 | 2002-06-18 | キンバリー クラーク ワールドワイド インコーポレイテッド | Neo nanoplast and microemulsion technology for ink and ink jet printing |
| AU5219299A (en) | 1998-07-20 | 2000-02-07 | Kimberly-Clark Worldwide, Inc. | Improved ink jet ink compositions |
| JP2003533548A (en) | 1998-09-28 | 2003-11-11 | キンバリー クラーク ワールドワイド インコーポレイテッド | Chelates containing quinoid groups as photopolymerization initiators |
| ES2195869T3 (en) | 1999-01-19 | 2003-12-16 | Kimberly Clark Co | NEW COLORS, COLOR STABILIZERS, INK COMPOUNDS AND IMPROVED METHODS FOR MANUFACTURING. |
| US6331056B1 (en) | 1999-02-25 | 2001-12-18 | Kimberly-Clark Worldwide, Inc. | Printing apparatus and applications therefor |
| US6294698B1 (en) | 1999-04-16 | 2001-09-25 | Kimberly-Clark Worldwide, Inc. | Photoinitiators and applications therefor |
| US6368395B1 (en) | 1999-05-24 | 2002-04-09 | Kimberly-Clark Worldwide, Inc. | Subphthalocyanine colorants, ink compositions, and method of making the same |
| US6211359B1 (en) * | 1999-08-17 | 2001-04-03 | Avl Medical Instruments | Triaza-cryptand and method of determining an alkali ion |
| US6962992B2 (en) | 2000-12-20 | 2005-11-08 | Molecullar Probes, Inc. | Crown ether derivatives |
| US7579463B2 (en) * | 2000-12-20 | 2009-08-25 | Life Technologies Corporation | Crown ether derivatives |
| FR2841468B1 (en) * | 2002-06-28 | 2004-09-10 | Oreal | USE FOR DYEING HUMAN KERATINIC FIBERS OF A CHROMOIONOPHORE AND / OR A FLUOROIONOPHORE, COMPOSITIONS AND METHODS FOR DYEING |
| US6997963B2 (en) * | 2002-06-28 | 2006-02-14 | L'oreal | Use of a chromoionophore and/or fluoroionophore for dyeing human keratin fibres, dyeing compositions and methods |
| BRPI0314042B8 (en) | 2002-09-06 | 2021-05-25 | Calando Pharmaceuticals Inc | cyclodextrin-based polymers for the delivery of covalently linked therapeutic agents |
| KR20050051686A (en) * | 2002-10-09 | 2005-06-01 | 인설트 테라페틱스, 인코퍼레이티드 | Cyclodextrin-based materials, compositions and uses related thereto |
| WO2005001016A1 (en) * | 2003-06-27 | 2005-01-06 | Interuniversitair Microelektronica Centrum (Imec) | Semiconductor cleaning solution |
| US7282349B2 (en) * | 2003-12-16 | 2007-10-16 | Kimberly-Clark Worldwide, Inc. | Solvatochromatic bacterial detection |
| DE102004059704B4 (en) | 2004-12-10 | 2012-07-05 | Scieva Gmbh | Spectroscopic detection of inorganic, organic or biological substances |
| WO2007131054A2 (en) * | 2006-05-03 | 2007-11-15 | Opti Medical Systems | Chromoionophore and method of determining sodium ions |
| WO2007131042A2 (en) * | 2006-05-03 | 2007-11-15 | Opti Medical Systems, Inc | Chromoionophore and method of determining potassium ions |
| DE102006023083A1 (en) * | 2006-05-16 | 2007-11-22 | Jacobs University Bremen Ggmbh | Determination of concentration changes |
| US20080176958A1 (en) | 2007-01-24 | 2008-07-24 | Insert Therapeutics, Inc. | Cyclodextrin-based polymers for therapeutics delivery |
| US8974814B2 (en) * | 2007-11-12 | 2015-03-10 | California Institute Of Technology | Layered drug delivery polymer monofilament fibers |
| CA2781669A1 (en) * | 2009-11-23 | 2011-05-26 | Cerulean Pharma Inc. | Cyclodextrin-based polymers for therapeutic delivery |
| CN102749312B (en) * | 2012-03-09 | 2014-06-25 | 华东师范大学 | Method for high selectivity recognition of acetate ions by applying perimidine onium anion receptor |
| WO2014055493A1 (en) | 2012-10-02 | 2014-04-10 | Cerulean Pharma Inc. | Methods and systems for polymer precipitation and generation of particles |
| US9360474B2 (en) | 2012-11-29 | 2016-06-07 | Opti Medical Systems, Inc. | Multi-layer device for selectively determining magnesium ion |
| US20150037899A1 (en) * | 2013-07-30 | 2015-02-05 | Water Lens, LLC | Borate detector composition and assay solution |
| JP6557894B2 (en) * | 2015-01-28 | 2019-08-14 | 新潟県 | Sake identity identification method and sake identification method |
| CN114539215B (en) * | 2022-01-05 | 2023-06-13 | 温州大学 | Merocyanine fluorescent probe, preparation method and application |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA966127A (en) * | 1970-05-12 | 1975-04-15 | Marilyn H. Bromels | Macrocyclic hetero imine complexing agents |
| US3789116A (en) * | 1970-12-09 | 1974-01-29 | Abbott Lab | Fluorescent labeled antibody reagent |
| US4005983A (en) * | 1972-10-16 | 1977-02-01 | Harald Dahms | Method and apparatus for colorimetric analysis |
| US3950135A (en) * | 1974-07-24 | 1976-04-13 | Massachusetts Institute Of Technology | Method of sepctral analysis using nmr shift reagents |
| US3992149A (en) * | 1975-02-18 | 1976-11-16 | Calspan Corporation | Colorimetric method for the analysis of residual anionic or cationic surfactants |
| US4013414A (en) * | 1975-05-05 | 1977-03-22 | General Electric Company | Indicator dye |
| IL49685A (en) * | 1976-05-31 | 1978-10-31 | Technion Res & Dev Foundation | Specific binding assay method for determining the concentration of materials and reagent means therefor |
| US4099918A (en) * | 1977-04-18 | 1978-07-11 | State Board Of Higher Education For And On Behalf Of The University Of Oregon | Proxyl nitroxides as spin labels |
-
1978
- 1978-10-02 DE DE19782842862 patent/DE2842862A1/en not_active Withdrawn
-
1979
- 1979-09-11 ZA ZA00794814A patent/ZA794814B/en unknown
- 1979-09-25 US US06/078,738 patent/US4367072A/en not_active Expired - Lifetime
- 1979-09-26 EP EP79103651A patent/EP0010615B1/en not_active Expired
- 1979-09-26 DE DE7979103651T patent/DE2967135D1/en not_active Expired
- 1979-09-26 AT AT79103651T patent/ATE8713T1/en not_active IP Right Cessation
- 1979-09-26 AU AU51233/79A patent/AU515023B2/en not_active Ceased
- 1979-10-01 SU SU792829004A patent/SU1135436A3/en active
- 1979-10-02 JP JP12649979A patent/JPS5548643A/en active Granted
- 1979-10-02 CA CA000336828A patent/CA1142841A/en not_active Expired
Also Published As
| Publication number | Publication date |
|---|---|
| EP0010615A1 (en) | 1980-05-14 |
| ZA794814B (en) | 1980-10-29 |
| AU515023B2 (en) | 1981-03-12 |
| JPH0240985B2 (en) | 1990-09-14 |
| US4367072A (en) | 1983-01-04 |
| JPS5548643A (en) | 1980-04-07 |
| EP0010615B1 (en) | 1984-07-25 |
| DE2842862A1 (en) | 1980-04-10 |
| ATE8713T1 (en) | 1984-08-15 |
| DE2967135D1 (en) | 1984-08-30 |
| SU1135436A3 (en) | 1985-01-15 |
| AU5123379A (en) | 1980-04-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA1142841A (en) | Process and a reagent for the determination of ions and of polar and/or lipophilic substances in fluids | |
| Verkman et al. | Synthesis and characterization of improved chloride-sensitive fluorescent indicators for biological applications | |
| CA1305133C (en) | Fluorogenic and chromogenic three-dimensional ionophores as selective reagents for detecting ions in biological fluids | |
| Nolan et al. | Selective Hg (II) detection in aqueous solution with thiol derivatized fluoresceins | |
| FI77937B (en) | TESTING INSTRUCTIONS FOR THE CONDITIONING OF THE CONDITIONS. | |
| Bhalla et al. | Rhodamine appended terphenyl: A reversible “off–on” fluorescent chemosensor for mercury ions | |
| Sheng et al. | A new colorimetric chemosensor for Hg2+ based on coumarin azine derivative | |
| JP2606721B2 (en) | Chromogen Cryptohemisferland and Its Use for the Detection of Electrolytes in Aqueous Test Samples | |
| CA1258672A (en) | Calcium complexing dyes and their use in analytical compositions, elements and methods | |
| JPS58134097A (en) | Potassium reagent and potassium ion analysis | |
| DE19639169A1 (en) | Redox-active compounds and their application | |
| Yun et al. | A novel benzophenone-based colorimetric chemosensor for detecting Cu^ 2+ Cu 2+ and F^-F | |
| JPS62261064A (en) | Analysis element and method for measuring magnesium ion | |
| US3880588A (en) | Diagnostic agent for detecting bilirubin | |
| He et al. | A fluorescent chemosensor for calcium with excellent storage stability in water | |
| CA1284932C (en) | Biological and analytical uses of phenalenone and benzphenalenone compounds | |
| US4992381A (en) | Method of detecting electrolytes in an aqueous test sample using chromogenic hemispherands and test device therefor | |
| EP0125554B1 (en) | Ion test means having a hydrophilic carrier matrix | |
| CN111269251A (en) | Fluorescent probe and test paper of nerve agent simulant and preparation method thereof | |
| US20070259438A1 (en) | Chromoionophore and method of determining calcium ions | |
| US4879243A (en) | Use of substituted ortho-quinones as electron transfer agents in analytical determinations | |
| EP0255679B1 (en) | Composition and method using substituted ortho-quinones as electron transfer agents in analytical determinations | |
| EP0140256B1 (en) | Composition for examining urobilinogen | |
| Peng et al. | ESIPT-based fluorescent enhanced probes prompted by methylated β-cyclodextrin for the detection of thiophenols | |
| US5177221A (en) | Chromogenic hemispherands and their preparation |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MKEX | Expiry |