LIGHT-SENSITIVE COMPOSITIONS FOR OXYGEN OPTRODES

Abstract: A composition comprising (a) an effective amount of a luminophore, (b) an oxygen-permeable and film-forming polymer and (c) a solvent for components (a) and (b), wherein (a1) the luminophore is selected from the group of platinum (II) porphyrins; and (a2) the polymers are selected from the group: (1) polystyrene-acrylonitriles having from 5 to 95 mol % structural units of acrylonitrile and from 95 to 5 mol % structural units of styrene; and (2) a hydrophobic homopolymer of an acrylate, methacrylate, acrylamide or methacrylamide, or a hydrophobic copolymer based on a monomer from the group acrylate, methacrylate, acrylamide and methacrylamide, and an olefinic comonomer. Support materials that have been coated with the solvent-free composition are suitable as optrodes for the luminescence-optical determination of oxygen, especially in physiological fluids.

Description: Light-sensitive compositions for oxygen optrodes The present invention relates to a composition comprising solvent, certain oxygen- permeable and membrane-forming polymers and a luminophore; to a support to which there is applied a layer comprising a certain oxygen-permeable and membrane-forming polymer and a luminophore; and to a method for the optical determination of oxygen. The quantitative determination of oxygen in gases and liquids by means of luminescence- optical methods and optrodes has recently become more important, especially in the field of diagnostics for the determination of the oxygen content in body fluids, for example blood. In Biosensors & Bioelectronics 7 (1991 ), pages 199 to 206, D. B. Pap ovsky et al. describe a fibre optic oxygen sensor having a polystyrene membrane in which platinum porphyrins are dissolved as fluorescence indicators. The luminescence produced by means of excitation radiation is extinguished, or reduced, by "quenching", in contact with oxygen, the extent of that extinction or reduction depending upon the amount of oxygen in the membrane. Thus, after calibration, the content of oxygen in an analyte sample can be measured by way of the decrease in the luminescence. A disadvantage of that system is a poor adhesion of the membrane to the support material. Moreover, the literature does not indicate whether it is possible, using fibre optic sensors and measurements in the evanescent field, to obtain a linear dependency of the concentration of oxygen upon the quotient \ (l 0 is luminescence in the absence of oxygen; I is luminescence in the presence of oxygen). In Applied Spectroscopy 46(8) (1992), pages 1266 to 1272, H.-Y Liu et al. mention platinum octaethylporphyrin as a fluorescence indicator in sol/gel-based membranes (polysiloxanes) for fibre optic sensors that are distinguished by a high permeability to oxygen. In some cases, a linear dependency of the quotient lo l upon the oxygen content is found (correlation coefficient maximum only 0.996), which is obtained, inter alia, by the specific selection of organic groups on the silicon atoms. Disadvantages of those sol/gel membranes are the decay behaviour of the excited phosphorescence indicator that is dissolved in the membranes, which can be described satisfactorily only by using a bi-exponential function, and the measuring sensitivity which, in some membranes, is associated with too sparing solubility of the luminophore in the polymer. Moreover, the optrodes are complicated to produce, which does not permit economic production on an industrial scale. US-A-5 55 149 describes fibre optic oxygen sensors in which the active layer consists of a c o po l yme r h avi n g s i loxa n e a n d u retha ne b loc k s , i n wh ic h a p lati n u m (tetraaminophenyl)porphyrin is bonded as fluorophore. It is mentioned that the permeability and solubility of oxygen can be influenced by the selection of the siloxane and urethane components. The covalent bonding of the fluorophore ensures its uniform distribution in the membrane. Functionalised fluorophores are difficult to produce and are therefore expensive, with the result that economic industrial mass production is not possible. In Sensors and Actuators A, 41 -42 (1994), pages 542 to 546, E. Singer et al. describe fibre optic optrodes for the detection of oxygen, the active layer of which consists of acrylate and methacrylate copolymers or cellulose acetate in combination with ruthenium(ll) tris(4,7-di- phenyl-1 ,10-phenanthrolin)perchlorate. The calibrating curves are non-linear, as a result of which a relatively large number of measuring points is necessary in the calibration in order to obtain precise measurements of the oxygen content of a measurement sample, which is felt to be too complicated. I n Anal Chem. 67 (1995), pages 88 to 93, P. Hartmann discloses planar optrodes comprising a glass or polyester support, to which there is applied a thin active layer of polystyrene that contains ruthenιum(ll) tris(4,7-diphenyl-1 ,10-phenanthrolin)perchlorate as fluorescence indicator. Those optrodes likewise do not demonstrate the desired ideal linear dependency of the quotient lo/l upon the partial pressure of the oxygen in the measurement sample, as is required in accordance with the Stern-Vollmer equation IQ/I = 1 +K*[O 2 ] for homogeneous solutions, wherein I is the luminescence in the presence of oxygen, l 0 is the luminescence in the absence of oxygen and K* is the quenching constant. In practice, it has proved advantageous to supplement the measured luminescence intensities, or rather the photoelectric voltages corresponding to them, in the Stern-Volmer equation by a constant C. Using the constant C, any possible background radiation, which may stem, for example, from reflections or scattering of the excitation light, or dark current effects are taken into account. There is a great need for optrodes for the detection of oxygen with which the calibration can be effected rapidly and in an uncomplicated manner using few, ideally two, measurements in a wide range of the O 2 partial pressure, for example from above 0 to 840 torr. The response times should be in the range of no longer than 40 seconds and it should be possible to regenerate the system rapidly and reuse it for new measurements with virtually no loss in measuring precision (identical luminescence intensities at identical concentrations of oxygen throughout the period of use) being able to occur, thus ensuring a long usable life It is also important that virtually no decomposition or other change occurs during storage. To ensure sufficient sensitivity, it is also advantageous for the quenching constant for the overall system in diagnostic methods (blood analysis) to be so adjusted that K is in the range of from 1 to 35 times 10 '3 torr *1 , preferably from 3 to 30 times 10 "3 torr '1 , especially from 5 to 25 times 10 '3 torr "1 . The obtained sensitivity (K) depends upon the duration of the excited state of the luminescence indicator in the absence of oxygen and upon the permeability of the polymer membrane to oxygen (product of the oxygen solubility and the diffusion coefficient of oxygen in the polymer membrane). The mentioned parameters influence the probability with which the luminescence of an excited indicator molecule will be extinguished by oxygen. In the case of relatively short durations, polymers having relatively high permeability to oxygen will therefore be selected in order to obtain the preferred sensitivity, and vice versa. The adjustment can be determined by the person skilled in the art by simple measurements based on the Stern-Vollmer equation It has now, surprisingly, been found that those high requirements can be fulfilled by combining platinum porphyπns as luminescence indicators with selected copolymers. The invention relates firstly to a composition comprising (a) an effective amount of a luminophore, (b) an oxygen-permeable and film-forming polymer and (c) a solvent for components (a) and (b), wherein (a1) the luminophore is selected from the group of platιnum(ll) porphyrms; and (a2) the polymers are selected from the group (1 ) polystyrene-acrylonitπles having from 5 to 95 mol % structural units of acrylonitrile and from 95 to 5 mol % structural units of styrene; and (2) a hydrophobic homopolymer of an acrylate, methacrylate, acrylamide or methacrylamide, or a hydrophobic copolymer based on a monomer from the group acrylate, methacrylate, acrylamide and methacrylamide, and an olefinic comonomer. Hydrophobic polyacrylamides generally contain 1 or 2 hydrocarbon radicals, especially alkyl groups having preferably from 1 to 20 carbon atoms, the total number of carbon atoms determining their hydrophobic property. Also included in the scope of the invention are co- polymers having at least two monomers from the group acrylate, methacrylate, acrylamide and methacrylamide. The acrylates and methacrylates are preferably those having a linear or branched CrC 2 oalkyl group in the ester group. The copolymers of component 2 contain preferably at least 5 mol %, especially at least 10 mol , more especially at least 20 mol % and very especially at least 40 mol % of an olefinic comonomer. Suitable comonomers are, for example, olefins, for example ethene, propene and butene, vinyl ethers, vinyl esters, vinyl acetals, vinyl chloride, vinylidene chloride, vinyl fluoride, acrylonitrile, methacrylonitrile and styrene. A preferred sub-group of copolymers of component 2 comprises those selected from the group: (a) a polymethacrylate or polyacrylate having from 95 to 70 mol % structural units of methyl methacrylate or methyl acrylate and from 5 to 30 mol % structural units of an acrylate or methacrylate having in the ester group an alkyl group that contains at least 3 carbon atoms; (b) a terpolymer having from 40 to 60 mol % structural units of methyl methacrylate or methyl acrylate, from 5 to 30 mol % structural units of an acrylate or methacrylate having in the ester group an alkyl group that contains at least 6 carbon atoms, and from 60 to 10 mol % structural units of acrylonitrile; and (c) a copolymer having from 30 to 45 mol % structural units of methyl methacrylate or methyl acrylate, from 20 to 5 mol % structural units of an acrylate or methacrylate having in the ester group an alkyl group that contains at least 6 carbon atoms, from 20 to 30 mol % structural units of acrylonitrile, and from 30 to 15 mol % structural units of vinyl acetate. The total of the mol % is always 100 mol %. The polymers used in the composition according to the invention are known or can be prepared in accordance with known processes by means of polymerisation of the monomers that form the basis of the structural units. The polystyrene-acrylonitriles (1 ) contain preferably from 15 to 85, especially from 20 to 70, more especially from 25 to 55 and very especially from 30 to 45 mol % structural units of acrylonitrile, the remainder of the structural units being of styrene. In the copolymers having structural units of an acrylate or methacrylate, the alkyl group of the ester group, provided it is not the methyl methacrylate or met acrylate itself, contains preferably from 2 to 18, especially from 3 to 14, more especially from 3 to 12 and very especially from 4 to 12 carbon atoms. The alkyl may be linear or branched. Examples thereof are n-butyl, isobutyl, n-pentyl, n-hexyl, 2-ethylbutyl, n-heptyl, 2- or 3-methylhexyl, 2- or 3-ethylpentyl, n-octyl, 2-ethylhexyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl and octadecyl. n-Butyl and 2-ethylhexyl are especially preferred. The polymethacrylates or polyacrylates (a) contain preferably from 10 to 25 and especially from 10 to 20 mol % structural units of an acrylate or methacrylate having an alkyl group that contains at least 4 carbon atoms, the remainder of the structural units being of methyl methacrylate or methyl acrylate. The terpolymers (b) contain preferably from 40 to 50 mol % structural units of methyl methacrylate or methyl acrylate, from 10 to 25 mol % structural units of an acrylate or methacrylate having in the ester group an alkyl group that contains at least 6 carbon atoms, and from 50 to 25 mol % structural units of acrylonitrile. The copolymer (c) contains preferably from 35 to 45 mol % structural units of methyl methacrylate, from 5 to 15 mol % structural units of an acrylate having in the ester group an alkyl group that contains at least 6 carbon atoms, from 30 to 20 mol % structural units of acrylonitrile, and from 30 to 20 mol % structural units of vinyl acetate. The molecular weight of the polymers may be, for example, from 10 000 to 5 000 000, preferably from 20 000 to 3 000 000, especially from 20 000 to 2 000 000 and more especially from 30 000 to 2 000 000 daltons. Better strengths of adhesion to the support material are often obtained with higher molecular weights. The compositions according to the invention can be prepared by dissolving components a) and b) in a solvent, where appropriate with heating. By means of the concentration and nature of the polymer, its molecular weight and the choice of solvent, the viscosity of the composition can be so adjusted that the desired layer thickness is achieved in the preparation of optrodes. Platinum porphyrins are known or can be prepared in accordance with known methods. They may be, for example, unsubstituted or substituted platιnum(ll) porphyrins or unsubstituted or substituted platinum(ll) benzoporphyrins. They may correspond, for example, to formula I, wherein R is H or C-i-Ciβalkyl, or C 3 -C β cycloalkyl, phenyl, pyridyl or phenyl-Cι-C 4 alkylene, each of which is unsubstituted or substituted by CrC 18 alkyl, CrC 18 alkoxy, R 3 -O-C(O)-, halogen, -CN or by -NO 2 ; R-i and R 2 are each independently of the other H, C C 12 alkyl or CrCι 2 alkoxy, or R-, and R 2 together form -CH 2 CHR 4 CHR 5 CH 2 -, -OCH 2 O-, -OCH 2 CH 2 O- or -CH=CH-CH=CH-; R 3 is C C 18 alkyl, C 3 -C β cycloalkyl, phenyl, benzyl, C Ci2alkylphenyl or C C 12 alkylbenzyl; and R 4 and R 5 are each independently of the other H, C-|-C 12 alkyl or C C 12 alkoxy; with the proviso that when R-i and R 2 together form -CH 2 CHR 4 CHR 5 CH 2 - , R is not substituted or unsubstituted phenyl or pyridyl. It is mentioned by way of explanation that the excluded compounds are not luminescent. The substituents in the cyclic radicals of R, for example cyclohexyl or phenyl, may be bonded in the 2-, 3- or 4-positιon. The radicals are preferably monosubstituted, with the substituent being bonded especially in the 4-position. When R in formula I is alkyl, it is preferably linear or branched C C^-, especially Cι-C β - and more especially Cι-C -alkyl. Examples thereof are methyl, ethyl and the isomers of propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl and octadecyl. Linear alkyl is also preferred. When R in formula I is cycloalkyl, it is preferably C -Cτcycloalkyl, especially C 5 - or C 6 - cycloalkyl. Examples thereof are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. When R in formula I is phenylalkyl, it is preferably benzyl or 1 -phenyleth-2-yl. In a preferred sub-group, R in formula I is H or is phenyl that is unsubstituted or substituted by C Cι 8 alkyl, C-,-Cι 8 alkoxy, R 3 -O-C(O)-, halogen, -CN or by -NO 2 , and R-- and R 2 together form -CH=CH-CH=CH-; R 3 bei n g C -, -C h alky I, C 3 -C β cycloalkyl, phenyl, benzyl, C-ι-C 12 alkylphenyl or C C 12 - alkylbenzyl. R 3 is preferably C C 12 alkyl, especially CrC 6 alkyl, it being possible for the alkyl to be linear or branched. R T and R 2 as alkyl and alkoxy may be linear or branched and are preferably C-ι-C 8 alkyl or C C e alkoxy, more preferably C t -C 6 alkyl or CrC 6 alkoxy and especially C C 4 alkyl or Cι-C 4 alkoxy. Examples thereof are methyl, ethyl, n-propyl and isopropyl, n-butyl, isobutyl and t-butyl, pentyl, hexyl, methoxy, ethoxy, n-propoxy and isopropoxy, n-butoxy, isobutoxy and t-butoxy, pentoxy and hexoxy. An especially preferred sub-group of the compounds of formula I comprises those wherein R is H, phenyl or C C 4 alkylphenyl, and each of R-j and R 2 is H, C C 8 alkyl or -CH=CH-CH=CH-. Another especially preferred sub-group of the compounds of formula I comprises those wherein R is H or C C 8 alkyl, and R-i and R 2 together form -CH 2 CHR 4 CHR 5 CH 2 - wherein each of R 4 and R 5 is H or C-,-C alkyl. Examples of platinum(ll) porphyrins are platinum(ll) tetrabenzoporphyrin, platinum(ll) tetraphenyltetrabenzoporphyrin , platinum(ll) octaethylporphyrin and platinum(ll) cyclohexenoporphyrin. Platιnum(ll) porphyrins are known or can be prepared in accordance with analogous processes, for example by reaction of PtCI 2 with the corresponding porphyrins in, for example, benzonitrile as solvent at elevated temperatures. The platinum(ll) cyclohexenoporphyrins so obtainable can be dehydrogenated to the corresponding platinum(ll) benzoporphyrins using organic dehydrogenating agents, for example chloranil or dichlorodicyanobenzoquinone in a solvent. The porphyrins themselves are known and can be converted in accordance with analogous methods, for example starting from isoindolines or isoindoiine derivatives and aldehydes R-CHO, to form a compound of the formula wherein R-. , R 2 and R are as defined for formula I, and those compounds are then dehydrogenated in a known manner using dehydrogenating agents, for example chloranil, dichlorodicyanobenzoquinone, or catalytically using noble metal catalysts, to the porphyrins of formula II (II). The composition according to the invention comprises an organic solvent or mixtures of such solvents comprising at least two different solvents. The solvents are preferably dipolar and aprotic. Examples of solvents are ethers (diethyl ether, dipropyl ether, dibutyl ether, ethylene glycol dimethyl or diethyl ether, diethylene glycol dimethyl ether, tetrahydrofuran and dioxane), N-dialkylated acid amides or N-alkylated lactams (dimethylformamide, dimethylacetamide, N-methylpyrrolidone, N-methyicaprolactam) , sulfones and sulfoxides (tetramethylene sulfone and tetramethylene sulfoxide), esters and lactones (methyl acetate, octyl acetate, butyrolactone, caprolactone, valerolactone), nitriles (acetonitrile, butyronitrile, benzonitrile, benzylnitrile), hydrocarbons (benzene, toluene, xylene) , halogenated hydrocarbons (methylene chloride, chloroform, carbon tetrachloride, 1 ,1 ,1 -trichloroethane, 1 ,1 ,2,2-tetrachloroethane, chlorobenzene) and ketones (acetone, methyl ethyl ketone, methyl isopropyl ketone). Suitable solvents have been mentioned hereinabove. The choice of solvents is guided essentially by the solubility properties of the polymers and luminophores. Effective amount" means a concentration of compound of formula I that is sufficient to detect the extinction of luminescence on contact with oxygen. The concentration of compound of formula I can be, for example, from 0.01 to 20 % by weight, preferably from 0.1 to 10 % by weight, especially from 0.5 to 5 % by weight and more especially from 0.5 to 3 % by weight, based on the amount of components a) and b). The invention relates also to a composition comprising a) a solid support material, to which there is applied an active layer of b) a composition according to the invention. The support materials may be transparent, translucent or opaque materials, with transparent materials being preferred. Examples of materials are plastics, glasses, minerals, metal and semi-metal oxides, metal nitrides and metal and semi-metal carbides. Preferred support materials are inorganic glasses and plastics. The surfaces of the support materials may be activated in order to improve the adhesion, for example by means of a plasma treatment, or adhesion promoters may be bonded to the surface. There may also be an adhesion-promoting layer between the surface and the active layer. The surface area of the support material can be in the range of, for example, mm 2 to cm 2 , preferably from 1 mm 2 to 10 cm 2 , especially from 2 mm 2 to 5 cm 2 , more especially from 5 mm 2 to 5 cm 2 . In order to carry out multiple measurements there may be arranged on the support several, for example from 2 to 30, preferably from 2 to 20, polymer layers that are separate from one another and have been produced by means of screen printing or writing processes. For the sensors, the glass wafers may be cut into strips. The sensor area effectively used can be, for example, from 0.8 to 20 mm 2 . The layer thickness of the composition comprising polymer and compound of formula I can be, for example, from 0.5 to 1000 μm, preferably from 0.5 to 200 μm, especially from 0.5 to 50 μm and more especially from 0.5 to 5 μm. The concentration of compound of formula I can be, for example, from 0.01 to 20 % by weight, preferably from 0.1 to 10 % by weight, especially from 0.5 to 5 % by weight and more especially from 0.5 to 3 % by weight, based on the amount of components a) and b). The composition according to the invention can be prepared in accordance with known coating processes, for example spreading, knife application, pouring, writing, for example curtain pouring process or especially spin-casting. The composition according to the invention is excellently suitable for the detection of oxygen when the active layer is in contact with an analyte, which may be gaseous or liquid. The composition is especially suitable for human and animal diagnostics. The invention relates also to a method of determining oxygen in an analyte, in which an oxygen-sensitive layer of an optical sensor, which layer contains a luminescence indicator and an oxygen-permeable polymer, is irradiated with light and luminescence radiation is produced, then the analyte is brought into contact with the layer and the reduction in the intensity of the luminescence radiation in dependence upon the oxygen content is detected, wherein the oxygen-sensitive layer comprises (a1 ) a luminophore selected from the group of platinum(ll) porphyrins; which is uniformly distributed in (a2) a copolymer selected from the group: (1 ) polystyrene-acrylonitriles having from 5 to 95 mol % structural units of acrylonitrile and from 95 to 5 mol % structural units of styrene; and (2) a hydrophobic homopolymer of an acrylate, methacrylate, acrylamide or methacrylamide, or a hydrophobic copolymer based on a monomer from the group acrylate, methacrylate, acrylamide and methacrylamide, and an olefinic comonomer. The preferences and preferred forms indicated hereinabove for the two compositions apply also to the method according to the invention. Suitable radiation sources for exciting the luminescence are, for example, UV lamps (halogen and xenon lamps), light-emitting diodes, lasers and diode lasers. It may be advantageous to select, by means of filters, light of the wavelength at which the luminescence dye has an absorption maximum. The excitation radiation can fall on the surface of the optrode perpendicularly or obliquely. The luminescence light emitted by the sensors can be collected, for example using a lens system, and then fed to a detector. There may be used as detectors photoelectron multipliers or photodiodes. The lens system may be so arranged that the luminescence radiation is measured through a transparent support. Advantageously, the radiation is deflected via a dichroic mirror and, by means of filters, tight of the wavelength at which the luminescence dye has an emission maximum is selected. The measurements are advantageously carried out during contact with the calibrating solutions or sample solutions or gas samples. For quantitative measurements, the sensors are calibrated using samples having a known oxygen content. The measurements can be carried out in fluorescence spectrometers which are commercially available. The oxygen detection can be carried out discontinuously in individual cells or continuously in flow-through systems; using the stop/flow method, it is possible to carry out a measurement several times or to measure different analytes in succession. Parallel measurements are also possible if multi-channel systems are used. The measuring temperature of the analytes can vary and, depending on the analyte medium, can be, for example, from -20°C to 200°C, preferably from 0 to 100°C and especially from 10 to 50°C. It is therefore possible without difficulty to take measurements in the physiological temperature range of approximately 37°C without damaging the optrodes. The measuring arrangement and the measuring conditions may influence the sensitivity, which, if desired, can be determined by the person skilled in the art by simple measurements. The invention relates also to the use of a composition comprising (a) an effective amount of a luminophore selected from the group of platinum(ll) porphyrins and (b) an oxygen- permeable film-forming copolymer selected from the group: (1 ) polystyrene-acrylonitriles having from 5 to 95 mol % structural units of acrylonitrile and from 95 to 5 mol % structural units of styrene; and (2) a hydrophobic homopolymer of an acrylate, methacrylate, acrylamide or methacrylamide, or a hydrophobic copolymer based on a monomer from the group acrylate, methacrylate, acrylamide and methacrylamide, and an olefinic comonomer; as the active layer of a sensor (optrode) for the luminescence-optical determination of oxygen. The preferences and forms indicated hereinabove for the composition according to the invention apply also to that use. The Examples which follow illustrate the invention in greater detail. A) Preparation of platinum(ll) tetracvclohexenoporphyrin Example A1 : Preparation of platinum(ll) cyclohexenoporphyrin A mixture of 1 .0 g (1 .9 mmol) of tetracyclohexenoporphyrin, 2 g of Cl2Pt(C 6 H 5 CN) 2 (4.2 mmol) and 200 ml of freshly distilled benzonitrile is heated under reflux, with stirring, for 2 hours. The mixture is then left to stand at room temperature overnight, and the precipitate that forms is filtered off. Drying is carried out overnight in vacυo at 100°C and 1.27 g (92 %) of a dark-red microcrystalline powder are obtained. Absorption spectrum in CHCI 3 , ^^(ε): 380 (266400), 501 (11000) and 536 (54900). 1 H-NMR (500 MHz, CHCI 3 ): 9.86 (4H, s, broad); 4.10 (16H, s, broad) and 2.50 (16H, s). Phosphorescence quantum yields: Φ 0 (in degassed CHCI 3 ): 0.421 Φ (in air-saturated CHCI 3 ): 0.0011. B) Preparation of coating compositions Examples B1 to B10: a) Preparation process for Examples B1 -B5: First, an approximately 0.1 % saturated solution of platinum(M) octaethylporphyrin in chloroform is prepared. 100 mg of styrene- acrylonitrile copolymer are then dissolved in 2 g of the dye solution (approximately 5 % polymer). The concentration of dye is approximately 2 % by weight, based on the polymer. b) Preparation process for Examples B6 to B10: 5 % by weight (based on the solution) of copolymer is dissolved in a dimethylacetamide solution saturated with platinum(ll) octaethylporphyrin . Table 1 : Composition of the styrene-acrylonitrile copolymers Example Stvrene (mol %) Acrvlonitrile (mol %) B1 91.7 8.3 B2 79.7 20.3 B3 71.6 28.4 B4 66.2 33.8 B5 58.5 41.5 B6 43.8 56.2 B7 37.2 62.8 B8 26.7 73.3 B9 18.0 82.0 B10 6.8 93.2 Example B1 1 : 200 mg of copolymer of 91.7 mol % styrene and 8.3 mol % acrylonitrile and 1 mg of platιnum(ll) tetrabenzoporphyrin are dissolved in 2 g of tetrahydrofuran. Example B12: A saturated solution of platinum(ll) tetracyclohexenoporphyrin in chloroform is prepared. 50 mg of copolymer of 58.5 mol % styrene and 41.5 mol % acrylonitrile are dissolved in 2 g of that solution. Examole B13: First, an approximately 0.1 % solution of platinum(ll) tetraphenyltetrabenzoporphyrin in chloroform is prepared. 100 mg of copolymer of 79.7 mol % styrene and 20.3 mol % acrylonitrile are then dissolved in 2 g of that dye solution (approximately 5 % polymer). The concentration of dye is approximately 2 % by weight, based on the polymer. Examples B14 to B19: First, an approximately 0.1 % solution of platinum(ll) octaethylporphyrin in chloroform is prepared. 100 mg of copolymer according to Table 2 are then dissolved in 2 g of that dye solution (approximately 5 % polymer). The concentration of dye is approximately 2 % by weight, based on the polymer. Table 2: Composition of the methacrylate copolymers corresponding to the monomer proportioi ns used for the polymerisation. Ex- Methvl methacrvlate 2-Ethvlhexvl methacrvlate n-Butvl methacrvlate (mol %) (mol %) (mol %) B14 95 5 B15 90 10 B16 85 15 B17 80 20 B18 95 5 B19 90 10 Example B20: First, an approximately 0.1 % solution of platinum(ll) tetraphenyltetrabenzoporphyrin in chloroform is prepared. 100 mg of copolymer of 85 mol % methyl methacrylate and 15 mol % 2-ethylhexyl methacrylate are dissolved in 2 g of that dye solution (approximately 5 % polymer). The concentration of dye is approximately 2 % by weight, based on the polymer. Examples B21 to B24: First, a saturated solution of platinum(ll) octaethylporphyrin in tetrahydrofuran is prepared. 220 mg of copolymer according to Table 3 are then dissolved in 2 g of that dye solution (10 % polymer). Table 3: Composition of the methacrylate copolymers corresponding to the monomer proportions used for the polymerisation. Example Methyl methacrylate Acrylonitrile (mol %) 2-Ethylhexyl acrylate (mol %) (mol %) B21 45 50 5 B22 45 45 10 B23 45 40 15 B24 45 35 20 Example B25: First, a saturated solution of platinum(ll) tetracyclohexeπoporphyrin in chloroform is prepared. 50 mg of copolymer of 45 mol % methyl methacrylate, 10 mol % 2-ethylhexyl acrylate and 45 mol % acrylonitrile are then dissolved in 2 g of that dye solution (2.5 % polymer). Example B26: First, a 0.1 % solution of platinum(ll) tetraphenyltetrabenzoporphyrin in chloroform is prepared. 100 mg of copolymer of 45 mol % methyl methacrylate, 20 mol % 2-ethylhexyl acrylate and 35 mol % acrylonitrile are then dissolved in 2 g of that dye solution (approximately 5 % polymer) . The concentration of dye is approximately 2 % by weight, based on the polymer. Example B27: First, a saturated solution of platinum(ll) octaethylporphyrin in tetrahydrofuran is prepared. 10 % by weight of copolymer of 40 mol % methyl methacrylate, 15 mol % 2-ethylhexyl acrylate, 25 mol % acrylonitrile and 20 mol % vinyl acetate are then dissolved in 2 g of that dye solution. The concentration of dye is approximately 1 % by weight, based on the polymer. Example B28: First, a saturated solution of platinum(ll) octaethylporphyrin in tetrahydrofuran is prepared. 10 % by weight of copolymer of 40 mol % methyl methacrylate, 10 mol % 2-ethylhexyl acrylate, 25 mol % acrylonitrile and 25 mol % vinyl acetate are then dissolved in 2 g of that dye solution. The concentration of dye is approximately 1 % by weight, based on the polymer. C) Preparation of sensors Example C1 : 150 μl of a solution according to Examples B1 to B10, B12, B14 to B19, B21 to B24, B27 and B28 are applied, under a solvent atmosphere, to a 2.5 cm -sized silanised glass plate by spin-casting at from 3000 to 6000 (usually 3500) revolutions/minute, and then the solvent is removed in a nitrogen stream at elevated temperature (80°C, at least 2 hours). The layer thicknesses are from 0.5 to 5 μm. In the case of the polymer solutions according to Examples B6 to B10, pre-drying is carried out immediately after application of the polymer solution in order to prevent the high-boiling solution from contracting on the glass plate. Example C2: 1 ml of a solution according to Examples B11 , B13, B20 and B26 is applied, under a solvent atmosphere, to a 36 cm 2 -sized silanised glass plate by spin-casting at from 6000 to 9000 revolutions/minute, and the solvent is then removed in a nitrogen stream at elevated temperature (80°C, at least 2 hours). The layer thicknesses are from 0.5 to 5 μm. In the case of the polymer solution according to Example 26, the solution is additionally diluted with chloroform to lower the viscosity. F) Application Examples Example F1 : Calibrations The support is so arranged that the excitation radiation falls on the active layer at an oblique angle. The radiation source used is a green light-emitting diode with the interposition of a shortpass interference filter (KIF, Schott-Schleifer AG) that has a maximum transmission (λ max ) at a wavelength of 533.5 nm and 50% transmission (X^a^) at a wavelength of 566.5 nm. A 645 DF 20 band filter (G+P Electronic AG) is used as the emission filter. The phosphorescence radiation is measured using a photodiode having an amplification of 10 11 . Moreover, the photoelectric voltage is reduced by means of a voltage distributor to such an Moreover, the photoelectric voltage is reduced by means of a voltage distributor to such an extent that the photoelectric voltage obtained for an oxygen-free measurement sample is less than 1 volt. There is used as measurement sample a sodium chloride-containing phosphate buffer [PBS, ionic strength 0.1 M (NaCI)], which, in each case, in the range from 0 to 840 torr has an oxygen content in 10 % steps. The measurement samples and also the entire measuring apparatus are brought to a temperature of 37°C. The measured photoelectric voltages, which are proportional to the corresponding luminescence intensities, are supplemented by a constant C that takes into account any possible background radiation, which may stem, for example, from reflections or scattering of the excitation light, or dark current effects. The constant C is calculated from the photoelectric voltages obtained at 0, 420 and 840 torr in accordance with the expanded Stern-Volmer equation (l 0 -C)/(l-C)= 1 + K*[OJ. All the measured photoelectric voltages are then corrected by that value for C and only then inserted into the relationship. The results are shown in Figures 1 to 3 The Figures demonstrate impressively the high degree of linearity of the calibration curves, for which correlation coefficients in the range from r=0 9997 to r=0.99997 were obtained. Figure 1 shows the calibration curves of sensors that have been coated with the coating compositions according to Examples B1 to B10. The oxygen partial pressure is plotted on tne X axis in increments of 100 torr, and the corrected relationship of the luminescence •ntensity in the absence and presence of oxygen l 0 -C/l-C is plotted on the Y axis in increments of 5 Figure 2 shows the calibration curves of sensors coated with the coating compositions according to Examples B14 to B19. Figure 3 shows the calibration curves of sensors coated with the coating compositions according to Examples B21 to B24 and B27 to B28. Measurement using a sensor and compositions according to Examples B1-B10: O 2 Phosphorescence intensity (mV) (torr) B 1 B 2 B 3 B 4 B 5 B 6 B 7 B 8 B 9 B 10 0 962.9 817.3 919.3 909.3 716.6 174.6 185.4 182.0 113.5 124.4 84 269.9 249.5 332.0 345.6 298.7 97.0 11 1.8 117.8 87.5 104.7 168 166.7 158.5 218.4 227.3 199.4 73.3 86.5 93.1 74.8 94.3 252 123.2 118.4 165.6 171.3 151.2 60.8 72.5 78.7 66.5 87.0 336 98.5 95.3 133.9 138.0 122.7 52.6 63.4 69.3 60.7 81.2 420 83.5 80.6 113.8 116.2 103.3 47.0 57.2 62.4 56.4 76.6 504 J 72.8 70.1 98.9 100.5 89.5 43.2 52.5 57.4 52.8 72.8 588 j 64.4 62.4 88.4 89.1 79.4 40.1 49.0 53.6 50.1 69.6 672 58.3 56.3 79.4 79.8 71.1 37.5 46.2 50.1 47.8 67.1 756 53.2 51.6 72.5 72.5 64.6 35.4 43.8 47.5 45.9 64.5 840 49.2 47.6 67.0 66.4 59.1 33.8 41.7 45.2 44.2 62.4 Measurement using a sensor and compositions according to Examples B14-B19 O 2 Phosphorescence intensity (mV) (torr) B 14 B 15 B 16 B 17 B 18 B 19 0 859.4 840.4 813.3 752.3 874.8 757.7 84 492.6 380.7 282.5 242.5 612.6 494.9 168 358.5 260.1 183.6 152.2 483.6 381.2 252 283.7 199.9 137.5 112.0 400.1 313.0 336 235.3 162.6 111.5 89.5 341.9 265.8 420 202.0 138.4 94.3 74.7 299.3 232.8 504 176.4 121.1 82.4 64.6 266.7 206.8 588 157.2 107.8 73.4 56.6 241.5 187.7 672 142.0 97.5 66.6 50.6 219.2 171.0 756 129.5 89.4 61.1 45.9 201.0 157.4 840 | 1 19.0 82.6 56.5 42.0 185.6 146.0 Measurement using a sensor and compositions according to Examples B21 -B24 and B27 and B28: o 2 Phosphorescence intensity (mV) (torr) B 21 B 22 B 23 B 24 B 27 B 28 0 515.9 577.3 861.6 704.6 904.2 754.0 84 382.6 323.2 362.0 207.7 283.0 339.0 168 31 1.2 241.9 255.1 140.6 187.0 238.1 252 263.4 197.7 202.4 1 10.3 141 .2 187.7 336 229.4 170.8 170.9 93.0 1 15.3 156.9 420 203.6 151 .1 149.6 81 .7 98.5 136.1 504 184.1 136.9 134.6 73.4 86.6 121 .1 588 168.2 125.9 123.1 67.2 77.2 109.1 672 155.0 1 1 7.0 1 14.0 62.5 70.0 100.1 756 143.7 109.9 106.8 58.8 64.2 92.6 840 134.1 j 104.0 101.0 55.7 59.6 86.5 Example F2: Examples using platinum(ll) tetracyclohexenoporphyrin (same apparatus as in Example F1) The support is so arranged that the excitation radiation falls on the active layer at an oblique angle. The radiation source used is a green light-emitting diode with the interposition of a shortpass interference filter (KIF, Schott-Schleifer AG) that has a maximum transmission (λ max ) at a wavelength of 533.5 nm and 50% transmission (λKarrt) a a wavelength of 566.5 nm. A 645 DF 20 band filter (G+P Electronic AG) is used as the emission filter. The phosphorescence radiation is measured using a photodiode having an amplification of 10 11 . Moreover, the photoelectric voltage is reduced by means of a voltage distributor to such an extent that the photoelectric voltage obtained for an oxygen-free measurement sample is less than 1 volt. There is used as measurement sample a sodium chloride-containing phosphate buffer [PBS, ionic strength 0.1 M (NaCI)] that has an oxygen partial pressure of 0, 80 and 160 torr. The measurement samples and also the entire measuring apparatus are brought to a temperature of 37°C. The result is given in the following Tables . Measurement using a sensor and compositions according to Examples B12 and B25' O 2 partial pressure Phosphorescence intensity (mV) (torr) B 12 B 25 0 448.2 139.9 80 232.2 99.7 160 168.3 82.7 Example F3: Measurements using platιnum(ll) tetraphenylporphynn using a channel cell The support is so arranged that the excitation radiation falls on the active layer at an oblique angle. The radiation source used is an orange light-emitting diode with the interposition of a shortpass interference filter (KIF, Schott-Schleifer AG) that has a maximum transmission (λ max ) at a wavelength of 487.0 nm and 50% transmission (λKgηt) at a wavelength of 607 7 nm An RG 9 longpass filter (Schott-Schleifer AG) is used as the emission filter. The phosphorescence radiation is measured using a photodiode having an amplification of 2*10 1 0 There is used as measurement sample a sodium chloride-containing phosphate buffer [PBS, ionic strength 0 1 M (NaCI)] that has an oxygen partial pressure of 0, 80 and 160 torr. The result is given in the following Tables Measurement using a sensor and compositions according to Example B1 O 2 partial pressure (torr) Phosphorescence intensity (V) 0 4.773 80 3.578 160 3.355 Example F4- Measurements using platinum (I I) tetraphenyltetrabenzoporphyrin using a channel cell (same apparatus as in Example F3). Measurement using a sensor and compositions according to Examples B13, B20 and B26: 0 2 partial pressure Phosphorescence intensity (V) (torr) B 13 B 20 B 26 0 1.417 1.821 3.103 80 0.771 1.219 2.505 160 0.614 0.982 2.256

Claims: What is claimed is: 1. A composition of (a) an effective amount of a luminophore, (b) an oxygen-permeable and film-forming polymer and (c) a solvent for the components (a) and (b), wherein (a1) the luminophore is selected from the group of platιnum(ll) porphyrins, and (a2) the polymers are selected from the group (1 ) polystyrene-acrylonitπles having from 5 to 95 mol % structural units of acrylonitrile and from 95 to 5 mol % structural units of styrene; and (2) a hydrophobic homopolymer of an acrylate , methacrylate, acrylamide or methacrylamide, or a hydrophobic copolymer based on a monomer from the group acrylate, methacrylate, acrylamide and methacrylamide, and an olefinic comonomer 2 A composition accordmg to claim 1 , wherein the hydrophobic polyacrylamides and polymethacrylamides generally contain 1 or 2 hydrocarbon radicals on the nitrogen atom 3 A composition according to claim 2, wherein the hydrocarbon radicals are alkyl groups 4 A composition according to claim 3, wherein the alkyl groups contain from 1 to 20 carbon atoms 5 A composition according to claim 1 , wherein the copolymers (2) are copolymers comprising at least two monomers from the group acrylate, methacrylate, acrylamide and methacrylamide 6 A composition according to claim 1 , wherein the acrylates (2) and methacrylates (2) are those having a linear or branched Cι-C 2C) alkyl group in the ester group 7 A composition according to claim 1 , wherein the copolymers (2) contain at least 5 mol % of an olefinic comonomer 8 A composition according to claim 1 , wherein the comonomers are olefins, vinyl ethers, vinyl esters, vinyl acetals, vinyl chloride, viπylidene chloride, vinyl fluoride, acrylonitrile, methacrylonitπle and styrene 9. A composition according to claim 1 , wherein the copolymers of component (2) are selected from the group: (a) a polymethacrylate or polyacrylate having from 95 to 70 mol % structural units of methyl methacrylate or methyl acrylate and from 5 to 30 mol % structural units of an acrylate or methacrylate having in the ester group an alkyl group that contains at least 3 carbon atoms; (b) a terpolymer having from 40 to 60 mol % structural units of methyl methacrylate or methyl acrylate, from 5 to 30 mol % structural units of an acrylate or methacrylate having in the ester group an alkyl group that contains at least 6 carbon atoms, and from 60 to 10 mol % structural units of acrylonitrile; and (c) a copolymer having from 30 to 45 mol % structural units of methyl methacrylate or methyl acrylate, from 20 to 5 mol % structural units of an acrylate or methacrylate having in the ester group an alkyl group that contains at least 6 carbon atoms, from 20 to 30 mol % structural units of acrylonitrile, and from 30 to 15 mol % structural units of vinyl acetate. 10 A composition according to claim 1 , wherein the polystyrene-acrylonitnles (1 ) contain preferably from 15 to 85 mol % structural units of acrylonitrile, the remainder of the structural units being of styrene 1 1 A composition according to claim 9, wherein the copolymers having structural units of an acrylate or methacrylate contain up to 18 carbon atoms in the alkyl group of the ester group, provided that it is not the methyl methacrylate or methyl acrylate itself. 12 A composition according to claim 9, wherein the polymethacrylates (a) or polyacrylates (a) contain preferably from 10 to 25 and especially from 10 to 20 mol % structural units of an acrylate or methacrylate having an alkyl group that contains at least 4 carbon atoms, the remainder of the structural units being of methyl methacrylate or methyl acrylate. 13 A composition according to claim 1 , wherein the terpolymers (b) contain from 40 to 50 mol % structural units of methyl methacrylate or methyl acrylate, from 10 to 25 mol % structural units of an acrylate or methacrylate having in the ester group an alkyl group that contains at least 6 carbon atoms, and from 50 to 25 mol % structural units of acrylonitrile. 14 A composition according to claim 1 , wherein the copolymer (c) contains from 35 to 45 mol % structural units of methyl methacrylate, from 5 to 15 mol % structural units of an acrylate having in the ester group an alkyl group that contains at least 4 carbon atoms, from 30 to 20 mol % structural units of acrylonitrile, and from 30 to 20 mol % structural units of vinyl acetate. 15. A composition according to claim 1 , wherein the molecular weight of the polymers is from 10 000 to 5 000 000. 16. A composition according to claim 1 , wherein the platinum porphyrin is an unsubstituted or substituted platinum(ll) porphyrin or an unsubstituted or substituted platinum(ll) benzoporphyrin. 17. A composition according to claim 1 , wherein the platinum porphyrin corresponds to formula I, wherein R is H or C-ι-Cι 8 alkyl, or C 3 -C 8 cycloalkyl, phenyl, pyridyl or phenyl-C C alkylene, each of which is unsubstituted or substituted by Cι-Cι 8 alkyl, C C 18 alkoxy, R 3 -O-C(O)-, halogen, - CN or by -NO 2 ; R-i and R 2 are each independently of the other H, C C 12 alkyl or C C 12 alkoxy, or R-i and R 2 together form -CH 2 CHR 4 CHR 5 CH 2 -, -OCH 2 O-, -OCH 2 CH 2 O- or -CH=CH-CH=CH-; R 3 is C C 18 alkyl, C 3 -C β cycloalkyl, phenyl, benzyl, Cι-C 12 alkylphenyl or Cι-Ci2alkylbenzyl; and R and R 5 are each independently of the other H, C Cι 2 alkyl or C C^lkoxy; with the proviso that when R-→ and R 2 together form -CH 2 CHFI 4 CHR5CH 2 -, R is not substituted or unsubstituted phenyl or pyridyl. 18. A composition according to claim 1 , wherein the compound of formula I is a compound wherein R is H, phenyl or Cι-C 4 alkylphenyl, and each of RT and R 2 is H, CrCβalkyl or -CH=CH-CH=CH-. 19. A composition according to claim 1 , wherein the compound of formula I is a compound wherein R is H or C C 8 alkyl, and R-→ and R 2 together form -CH 2 CHR CHR 5 CH 2 -, each of R 4 and R 5 being H or d-C 4 alkyl. 20. A composition according to claim 1 , wherein the compound of formula I is platinum(ll) tetrabenzoporphyrin, platinum(ll) tetraphenyltetrabenzoporphyrin, platinum (II) octaethylporphyrin or platinum(ll) cyclohexenoporphyrin. 21. A composition according to claim 1 that comprises as solvent ethers, N-dialkylated acid amides or N-alkylated lactams, sulfones and sulfoxides, esters and lactones, nitrites, hydrocarbons, halogenated hydrocarbons or ketones. 22. A composition according to claim 1 , wherein the amount of compound of formula I is from 0.01 to 20 % by weight, based on the amount of components a) and b). 23. A composition comprising a) a solid support material to which there is applied an active solvent-free layer of b) a composition according to claim 1. 24. A composition according to claim 23, wherein the support material is a transparent, Translucent or opaque material. 25. A composition according to claim 23, wherein the support material is an inorganic glass or plastics material. 26. A composition according to claim 23, wherein the layer thickness of the composition comprising polymer and compound of formula I is from 0.5 to 1000 μm. 27. A method of determining oxygen in an analyte, in which an oxygen-sensitive layer of an optical sensor, which layer contains a luminescence indicator and an oxygen-permeable polymer, is irradiated with light and luminescence radiation is produced, then the analyte is brought into contact with the layer and the reduction in the intensity of the luminescence radiation in dependence upon the oxygen content is detected, wherein the oxygen-sensitive layer comprises: (a1 ) a luminophore selected from the group of platinum(ll) porphyrins; which is uniformly distributed in: (a2) a copolymer selected from the group: (1) polystyrene-acrylonitriles having from 5 to 95 mol % structural units of acrylonitrile and from 95 to 5 mol % structural units of styrene; and (2) a hydrophobic homopolymer of an acrylate, methacrylate, acrylamide or methacrylamide, or a hydrophobic copolymer based on a monomer from the group acrylate, methacrylate, acrylamide and methacrylamide, and an olefinic comonomer. 28. The use of a composition comprising (a) an effective amount of a luminophore selected from the group of platinum(ll) porphyrins; and (b) an oxygen-permeable and film-forming copolymer selected from the group: (1 ) polystyrene-acrylonitriles having from 5 to 95 mol % structural units of acrylonitrile and from 95 to 5 mol % structural units of styrene; and (2) a hydrophobic homopolymer of an acrylate, methacrylate, acrylamide or methacrylamide, or a hydrophobic copolymer based on a monomer from the group acrylate, methacrylate, acrylamide and methacrylamide, and an olefinic comonomer; as the active layer of a sensor (optrode) for the luminescence-optical determination of oxygen.

Inventors: BERGER JOSEPH, SINGER ERIS, BERGER, JOSEPH, SINGER, ERIS
Applicants: BERGER JOSEPH, CIBA GEIGY AG, SINGER ERIS, BERGER, JOSEPH, NOVARTIS AG, SINGER, ERIS
Application year: 1997
Main classification: G01N31
IPC codes: C07D487, G01N21, G01N31