Abstract: This invention provides (1) a reagent for endotoxin assay which comprises aprotinin and a limulus amebocyte lysate reagent, (2) a kit for endotoxin assay which comprises the limulus amebocyte lysate reagent and a reagent containing aprotinin, (3) a method for assaying endotoxin in a sample using the limulus amebocyte lysate reagent in which aprotinin is added to the lysate reagent and/or the sample, (4) a method for assaying endotoxin in a serine protease-containing sample using the limulus amebocyte lysate reagent in which the sample is allowed to contact with an aprotinin-immobilized insoluble carrier in advance of endotoxin assay, (5) a carrier for pretreating a serine protease-containing sample on which aprotinin is immobilized, (6) a method for inhibiting factor G activation in which aprotinin is added to the limulus amebocyte lysate reagent and (7) a factor G activation inhibitor which comprises aprotinin as an active ingredient.

Abstract: Provided is a method of determining the amount of cholesterol in high-density lipoprotein (HDL), which comprises reacting an HDL-containing sample with cholesterol esterase and cholesterol oxidase or cholesterol dehydrogenase in the presence of a reagent for aggregating lipoproteins except HDL, and determining the amount of hydrogen peroxide or reductive co-enzyme formed therein.

Abstract: The present invention provides a reagent for determining (1.fwdarw.3)-.beta.-D-glucan comprising a lysate substantially free from any endotoxin-sensitive factor, which is obtained by contacting limulus amebocyte lysate optionally containing dextran with a polyamide or cellulose insoluble carrier, which makes it possible to rapidly and easily determine at a high accuracy (1.fwdarw.3)-.beta.-D-glucan of mycotic origin contained in the body fluid such as blood or urine.

Abstract: It was found that atopic allergens have enzymatic properties, in particular the properties to hydrolyze amide and/or ester linkages. These properties may be used for various purposes, e.g. for analysis of samples, standardization of pharmaceutical compositions and also for the preparation of the allergens in a pure form.

Abstract: A composition, method and test device for determining the presence of leukocytes, esterase or protease in a test sample are disclosed. The composition comprises an ester which is hydrolyzed in the presence of leukocyte, esterase, or protease to form a reaction product which couples with a diazonium salt to produce a detectable color change. The composition further comprises the salt of an alkaline earth metal which stabilizes the composition during manufacture and prevents the occurrence of false or background color change due to reactivity of the diazonium salt in the absence of leukocyte, esterase or protease.

Abstract: Enzyme activity is measured promptly with a high accuracy by introducing an enzyme, the activity of which is to be measured, into a column comprising a hollow tube packed with a filler comprising a support and a substrate that can be recognized by the enzyme, which is immobilized on the support, and measuring the amount of the obtained decomposition product of the substrate.

Abstract: The invention provides novel DNA and peptide sequences encoding a family of phospholipase A.sub.2 enzymes, with specific activities of approximately 20 .mu.mol/min/mg in the mixed micelle assay. These enzymes are useful in methods for detecting the anti-inflammatory potential of various chemical agents. The invention also details novel methods for determining such potential using the novel sequences, methods for making the novel peptides, and methods for developing new anti-inflammatory drugs.

Abstract: By contacting a peptide derivative or a protein having a property of binding to endotoxin (ET) to inhibit the activity of ET, and at least one surfactant with a sample containing ET, the activity of ET can be inhibited, and even when an ET analogous substance having ET activity is present in the thus treated sample, it can be measured without influence of ET.

Abstract: Pyrroloquinoline quinone-dependent glycerol dehydrogenase is provided which requires no surfactant for its solubilization and stabilization. The glycerol dehydrogenase has the following properties:(a) catalyzing the following reaction:glycerol+electron acceptor.fwdarw.glyceraldehyde+reduced electron acceptor,(b) optimum pH: pH 8-9,(c) pH stability: stable at pH 7-11,(d) optimum temperature: 20.degree.-25.degree. C,(e) thermal stability: stable at a temperature of 30.degree. C or less for 10 minutes at a pH of 7.0,(f) molecular weight: 70,000 by gel filtration and SDS polyacrylamide gel electrophoresis,(g) bound to pyrroloquinoline quinone as a prosthetic group,(h) soluble and stable without the need of an ionic or non-ionic surfactant.

Abstract: The invention describes biological assays in which hydrogen peroxide is used as an oxidizing agent, or wherein hydrogen peroxide is used to oxidize a dye or other intermediate to generate a detectable species. The stability of the hydrogen peroxide in the presence of at least one other enzyme which decomposes hydrogen peroxide is enhanced by the addition of a suitable inhibitor for the enzyme and the inhibitor does not substantially inhibit enzymes used in the assay. When catalase is the enzyme to be inhibited, catalase inhibitors that can be used in the biological systems include hydroxylamine sulfate. The enzyme inhibitor can be incorporated in an integral analytic device.

Abstract: A method is provided for diagnosing of senile dementia of the Alzheimer type (Alzheimer's disease) by measuring acetylcholinesterase (AChE) activity in ocular fluids and determining if such AChE activity is elevated over that found in ocular fluids of patients who do not have Alzheimer's disease. A level of AChE activity in the ocular fluid of a patient less than 30% higher than the level of AChE activity in the ocular fluids of a significant number of age-matched controls signifies the absence of Alzheimer's disease. A level of AChE activity in the ocular fluid of a patient at least about 35% higher than the level of AChE activity in the ocular fluids of a significant number of age-matched controls signifies the presence of Alzheimer's disease.

Abstract: A multilayer analysis element for determination of total cholesterol is prepared having a light-transmissive water-impermeable support, at least one hydrophilic polymer layer on said support and a spreading layer on said hydrophilic polymer layer(s), and containing in one or more of the layers:(a) at least one enzyme having cholesterol esterase activity,(b) cholesterol oxidase,(c) peroxidase,(d) a color reagent composition, which in the presence of hydrogen peroxide and the peroxidase, produces a color change(e) at least one bile acid compound selected from the group consisting of bile acids, bile acid derivatives and salts of said acids and derivatives, and(f) an alkyl phenoxy polyethoxy ethanol containing in the alkyl group 1 to 20, preferably 7 to 10 carbon atoms, and a polyoxyethylene chain composed of at least 16, preferably 30 to 60 oxyethylene units.

Abstract: Chemiluminescence-based assays that detect or quantify enzymes that catalyze the hydrolysis of indoxyl esters are provided. The assays are based on the hydrolysis of indoxyl esters by enzymes of interest, such as alkaline phosphatase and others that are used as labels in immunoassays or nucleic acid hybridization reactions, or are present in body fluids. The assays include the steps of reacting a test sample with an indoxyl ester and, then, immediately or within a short time, typically less than about fifteen minutes, measuring the resulting chemiluminescence. The resulting chemiluminescence may be amplified by adding a chemiluminescence-amplifying reagent, such as horseradish peroxidase or lucigenin to the reaction.

Abstract: The present invention provides compounds that function as hydrolytic enzyme inhibitors (inactivators) and substrates. These compounds are useful in assays to detect and measure levels of hydrolytic enzyme activity and are more particularly useful in treatment regimens for various disease states and conditions implicating the underlying specific hydrolytic enzyme. Examples of hydrolytic enzymes include, but are not limited to, phospholipases, lipases, esterases, proteases, etc.

Abstract: A dye is covalently bound to a polymeric film, especially a polyhydric polymer, for assays and other purposes. The dye may be one which, when it comes into contact with hydrogen peroxide, changes color to indicate the presence of hydrogen peroxide. This dyed film may be used in qualitative or quantitative assays. This method chemically immobilizes dyes on support matrices with much higher yields of immobilized dye than has heretofore been possible. The covalently immobilized dye may be immobilized on solid matrix particles and combined with a free-flowing dye component to form a two component dye system. By combining a dyed film-former with a film-opener, the amount of dye available for assay is greatly enhanced. This provides a dye system which can be used to detect and measure quantitatively, accurately and precisely high levels of hydrogen peroxide. These high levels of hydrogen peroxide may result from the enzyme-mediated decomposition of various analytes from undiluted whole blood samples.

Abstract: The invention relates to polymeric compositions useful in analyte determination. The compositions contain a polymer, a reagent system for analyte determination, and an extender. The last component alleviates tackiness in the composition, and thus reduces damage in preparation of test apparatus. Mica is the particularly preferred extender.

Abstract: Enzymatically clearable chemiluminescent 1,2-dioxetane compounds capable of producing light energy when decomposed, substantially stable at room temperature before a bond by which an enzymatically clearable labile substituent thereof is intentionally cleaved, are disclosed. These compounds can be represented by the formula: ##STR1## wherein: X and X.sup.1 each represent, individually, hydrogen, a hydroxyl group, a halo substituent, an unsubstituted lower alkyl group, a hydroxy (lower) alkyl group, a halo (lower) alkyl group, a phenyl group, a halophenyl group, an alkoxyphenyl group, a hydroxyalkoxy group, a cyano group or an amide group, with at least one of X and X.sup.1 being other than hydrogen; and R.sub.1 and R.sub.2, individually or together, represent an organic substituent that does not interfere with the production of light when the dioxetane compound is enzymatically cleaved and that satisfies the valence of the dioxetane compound's 4-carbon atom, with the provisos that if R.sub.1 and R.sub.

Abstract: Aryl N-alkylacridanthiocarboxylate compounds which produce chemiluminescence. The compounds produce light with peroxide and peroxidase. The compounds are used as a substrate in assays for various analytes.

Abstract: A method of assay for an enzyme catalysing the release of long chain fatty acids or, conversely, for an ester substrate for such enzymes is applicable to clinical samples and comprises removing substantially all the albumin from the clinical sample, incubating the sample after albumin removal with, for enzyme assay, an ester substrate or, for substrate assay, the enzyme, under conditions effective to release a fatty acid, causing the fatty acid to bind to a fatty acid binding protein and assaying the binding of the fatty acid to the binding protein.

Abstract: A test device for determining the presence of leukocyte cells, esterase or protease in a test sample is disclosed. The test device comprises a carrier matrix having a reagent composition incorporated therein, wherein the reagent composition comprises a lactate ester having the structure ##STR1## wherein A is an alcohol blocking group, and wherein B--O-- is a residue of compound B--OH and provides a detectable response when the lactate ester is hydrolyzed; and a buffer. In a preferred embodiment, the lactate ester has the structure ##STR2## wherein X is O, S or NR.sup.2, R is aryl or lower alkyl, R.sup.1 is hydrogen or lower alkyl, and R.sup.2 is hydrogen, lower alkyl or aryl.

Abstract: A method of inhibiting the function of .beta.-protein, particularly enzymatic activity, such as esterase (cholinesterase) or proteinase activity, is to contact .beta.-protein with a compound which inhibits such enzymatic activity. Examples of such inhibitors are para-amidinophenylmethanesulfonyl fluoride and Ebelactone A, which inhibit the esterase activity of amyloid precursor protein to a greater extent than the esterase activity of acetylcholinesterase.

Abstract: N-alkylacridan carboxylic acid derivative compounds (I) are used to generate chemiluminescence by the action of a peroxidase enzyme and an oxidant. The compounds I are useful in assays of all types.

Abstract: The present invention provides a kinetic or rate method for the determination of cholesterol in free or bound form with of a cholesterol oxidase enzyme obtained from Bacterial strain NRRL B-18713 which inherently has a high Michaelis and Menton, Km, of about 10-2 to 10-3 M for cholesterol and optionally, of cholesterol esterase. The method can measure oxygen consumption, hydrogen peroxide or cholestenone formed. The present invention also provides a test composition for the kinetic determination of cholesterol, comprising cholesterol oxidase with an inherently high Km of about 10-2 to 10-3 M for cholesterol and optionally cholesterol esterase and a system for the determination of hydrogen peroxide, cholestenone or oxygen consumption.

Abstract: A composition and method for determining the presence of leukocyte cells, esterase or protease in a test sample are disclosed. The composition contains a lactate ester having the structure ##STR1## wherein A is an alcohol blocking group, and wherein B-O- is a residue of compound B-OH and provides a detectable response when the lactate ester is hydrolyzed; and a buffer. One class of lactate ester has the structure ##STR2## wherein X is O, S or NR.sup.2, R is aryl or lower alkyl, R.sup.1 is hydrogen or lower alkyl, and R.sup.2 is hydrogen, lower alkyl or aryl.

Abstract: A non-radioactive, spectrophotometric, microtiter plate assay for human cystolic phospholipase A.sub.2 (cPLA.sub.2) is described. The assay utilizes a novel synthetic thiol-phospholipid analog as a substrate. In one embodiment, the substrate is a phosphatidylcholine derivative with an arachidonoylthioester in the sn-2 position and an alkenyl-ether or alkenyl-ether in the sn-1 position. The alkyl-ether and the alkenyl-ether in the sn-1 position of the substrate ensures that the assay will only measure cPLA.sub.2 activity and will not be complicated by metabolism of the lysophospholipid product by the enzyme's and lysophospholypase activity.

Abstract: A cocaine esterase has been isolated from a strain of the bacteria Pseudomonas maltophilta, The cocaine esterase catalyses the debenzoylation of cocaine, This reaction may be used in the detection of cocaine. The enzyme may be incorporated into sensors for this purpose. The cocaine esterase is preferably obtainable from Pseudomonas sp. NCIMB 40427. It catalyzes the debenzoylation of cocaine, has a molecular weight in the unaggregated form of about 120,000 daltons as determined by gel filtration, has esterase activity specifically at the benzoate ester linkage of cocaine, separates at a major band of Rf about 0.2 on PAGE in its aggregated form, and it is completely inhibited by 1 mM phenylmethylsulphonyl fluoride but ineffectively inhibited by 1 mM eserine, each determined at 30.degree. C. with respect to 2 mM cocaine as substrate.

Abstract: A method for the estimation of a salicylate in a sample is characterized by enzymatically converting the salicylate to a catechol by the action of a salicylate mono-oxygenase enzyme on the salicylate in the presence of a reduced pyridine nucleotide, reacting the catechol with a compound selected from compounds of formula I or amine or phenolic compounds of formulae II, III or IV ##STR1##

Abstract: A method for collecting an aerosolized therapeutic polypeptide of interest, such as recombinant human deoxyribonuclease I (rhDNase), is provided which enables a determination as to the effect aerosolization has on the activity and integrity of the polypeptide. An aerosol of the polypeptide is generated using a nebulizer, for example, and the polypeptide is collected in an inert filter, such as a sintered glass filter. To increase the amount of polypeptide collected, the aerosol is preferably mixed with pre-humidified dilution air at a temperature between about 40.degree. and 55.degree. C. The collected polypeptide is subjected to biochemical activity and integrity analysis compared to the activity and integrity of the control polypeptide which has not been aerosolized.

Abstract: A dye couple, comprising 3-methyl-2-benzothiazolinone hydrazone (MBTH) and 8-anilino-l-naphthalenesulfonate (ANS), is used as an indicator in a reaction cascade producing a strong oxidizing agent, such as hydrogen peroxide or other peroxides or perborates. The strong oxidizing agent reacts with the dye couple to produce a blue dye stuff. The MBTH-ANS dye couple exhibits strong and flat spectral absorption at the region of about 600 to 650 nm. This region is free of blood color interference, which enables one to measure glucose and other analytes that react with an oxidase enzyme to produce the strong oxidizing agent, through the use of LED optics, accurately without much optic calibration. Further, the MBTH and ANS are very soluble in aqueous solution, yet become insoluble upon oxidative coupling. The poor solubility minimizes dye fading, thus providing a stable endpoint.

Abstract: A method is provided for measuring the net HDL cholesterol in the blood. The LDL and VLDL moieties are precipitated out and the remaining cholesterol content of the supernatant is measured. The free cholesterol content of the supernatant is separately measured and substracted from the total to obtain net HDL cholesterol. This value serves as a useful indicator for diagnosing vascular disease. A diet supplement and method for raising serum HDL is provided.

Abstract: The present invention relates to a method of determining, assaying and identifying strains of Candida and pathogenic strains belonging to species Torulopsis glabrata by means of chromogenic substrates of monoamino acid or peptide type without the isolation of the strains to be identified.

Abstract: A method of quantitative assay for a substrate such as triglyceride capable of undergoing enzymatic hydrolysis to release long chain fatty acids in an albumin-containing clinical sample comprises incubating the albumin-containing clinical sample with an enzyme such as lipase which acts upon the substrate to be assayed under conditions effective to release fatty acid therefrom, causing the fatty acid thus released to bind to a fatty acid binding protein (FABP) and assaying the binding of the fatty acid to the FABP.

Abstract: Methods and compositions are disclosed for determining a peroxidatively active substance (PAS). The methods comprise the step of detecting a fluorescent signal produced upon cleavage of a compound of the formula F-L-Q, wherein F is a fluorescer capable of producing the signal, Q is a quencher capable of quenching the signal when linked to F, and L is a bond, or a linking group having a bond, wherein the bond is capable of being cleaved by a reaction of the PAS with a substrate of the PAS and a hydrogen donor wherein the cleavage of the bond substantially reduces the quenching. The methods have application in a wide variety of systems including assays and improved assays for analytes. Also disclosed are kits for conducting the methods and improvements in accordance with the present invention.

Abstract: The present invention provides an accurate, rapid and precise methodology for assessing maturity of the lungs in a fetus prior to birth, especially for premature fetuses having a gestation period of 37 weeks or less. The methodology quantitatively measures a specific phosphoglyceride, dipalmitoyl phosphatidyl choline, in samples of amniotic fluid for the assessment of fetal lung maturity. The process enzymatically cleaves such phophoglycerides and preferably detects the resulting diacylglycerols by HPTLC-reflectance spectrodensitometry.

Abstract: Subject matter of the invention is a new method of detecting a substance with hydrolase activity in a sample, characterized in that the sample is brought into contact with an indicator enzyme, which is covalently bound to an insoluble carrier material and can be rendered soluble by the hydrolase activity, in that the cleaved off indicator is separated and in that its enzymatic activity is determined and test means for its implementation.

Abstract: This invention is a method for measuring the ability of a human to absorb cholesterol. The method uses two different cholesterol tracers, the first injected into the blood stream and the second ingested by the human subject. After a waiting period a blood sample is taken from the human subject and analyzed to determine percent cholesterol absorption based on the actual amounts of the two naturally occurring, metabolically stable cholesterol tracers in the blood. The method of this invention is useful in identifying human subjects as high cholesterol absorbers and thereafter administering therapeutic agents to the subject that inhibit the absorption of cholesterol.

Abstract: A rapid method of determining the efficacy of a sterilization cycle, and an indicator adapted to perform such method, comprising subjecting to the sterilization cycle a source of active enzyme having activity which correlates with the viability of a microorganism commonly used to monitor sterilization, and incubating the enzyme source, following the completion of the sterilization cycle, with an effective amount of a substrate system capable of reacting with any residual active enzyme to produce a detectable enzyme-modified product.

Abstract: Low density lipoproteins are directly determined in fluid samples by selectively precipitating low density lipoproteins from the sample by forming clusters, selectively consuming the high density and very low density lipoproteins, and resolubilizing the low density lipoproteins for direct determination thereof. The clusters are formed by treating the fluid sample with a mixture of a polyanionic compound, a divalent metal, and a nucleating agent. The clusters are redissolved and assayed for low density lipoproteins.

Abstract: A laminated assay device for use in determining the presence or amount of an analyte in a sample involving a bibulous assay strip having a sample application zone, a reagent zone and a pair of liquid impervious barriers defining a measurement zone extending from and in fluid communication with the application zone. The measurement zone has a volume measuring the amount of sample required for the assay. A substantially nonabsorbent first supporting film extends across and is laminated to the front surface of the assay strip, while a substantially nonabsorbent second supporting film extends across and is laminated to the back surface of the assay strip. The assay strip is impregnated with the members of a signal producing system which, upon reaction with a component in the sample, produce a detectable signal.

Abstract: The invention refers to a method for determining the relative amounts of all cholesterol-containing lipoproteins in body fluids comprising electrophoretically separating the lipoproteins of an aliquot of body fluid on a thin layer carrier matrix, incubating the carrier matrix, containing the separated lipoproteins with cholesterol esterase and cholesterol dehydrogenase, forming a provable complex, and determining the relative amounts of the different lipoprotein classes2. The new method makes it possible to simultaneously determine HDL-, LDL-, VLDL- and LP (X)-cholesterol in body fluids with a high accuracy even at small concentrations. The thin layer matrices obtained electrophoretically, are very easy to handle and to record.

Abstract: A method of assaying for acyl-L-carnitines including short chain acyl-carnitines including acetyl-L-carnitine and propionyl-L-carnitine in a substance, comprises subjecting a sample of the substance to be analyzed to an enzymatic hydrolysis using an acyl-carnitine esterase. The esterase is produced by Alcaligenes sp. FERM BP-2570 and it has substrate specificity for acyl-L-carnitines including short-chain acyl-L-carnitines. In addition the esterase demonstrates substrate specificity for acetyl-L-carnitine and propionyl-L-carnitine. The enzyme facilitates the hydrolysis reaction of one mole each of the acyl-L-carnitines with one mole of water in which to form one mole each of the corresponding fatty acid and L-carnitine. The amount of the fatty acid and L-carnitine formed is determined by this method.

Abstract: The invention provides a method for supplying an enzyme substrate to a membrane-based, enzyme-linked reaction, comprising providing an open pore, high liquid retention capacity material impregnated with a predetermined amount of a substrate for the enzyme; and contacting the material with a membrane containing the enzyme-linked reaction under conditions which permit diffusion of the enzyme substrate to sites on the membrane containing the enzyme linked reaction.

Abstract: A single system reagent for the determination of lipase, formulated as an emulsion comprising a lipase-active fatty acid source; a first lipase activator comprising colipase; a second lipase activator; a lipoprotein lipase inhibitor; an emulsion stabilizer comprising Triton X100; a buffer; and an antiprecipitant.

Abstract: A method, reagent mixture and test kit are provided for testing a urine sample for the presence of bacterial or somatic cells, by reacting a urine sample with a catalase-free alkaline protease enzyme and a detergent compatible therewith, so as to disrupt any such cells present in the sample and release active catalase therefrom. The presence of catalase is detected by the formation of foam upon the addition of H.sub.2 O.sub.2. The reaction of the sample with the protease enzyme and the detergent is conducted in the presence of: (a) a buffer providing for a pH of about 8.5 to about 9; and optionally (b) one or more additional solutes; the total concentration of components (a) and (b), when present, being from about 0.02M to about 0.4M.

Abstract: The present invention relates to chromogenic compounds which are represented by the general Formula (I): ##STR1## wherein R.sub.1 is a sugar group, ester group, hydrocarbyl group, phosphate group, sulfate group or a salt thereof, with the proviso that R.sub.1 is other than .beta.-D-glucuronic acid or .beta.-D-galactopyranoside, R.sub.2 is H or hydrocarbyl group containing 1 to about 5 carbon atoms, X is Cl or H, and Y is Cl or H. The present invention further relates to a method for quantitatively identifying and differentiating a first biological material having enzyme specificity for a first chromogenic compound as represented by Formula (I) above, and a second biological material having enzyme specificity for a second chromogenic compound.

Abstract: Hydroxy coumarin ester enzyme substrates can be stored in a strong base at pH above 11 to stabilize the substrate against premature hydrolysis of the ester bond and to reduce the revel of contaminating hydroxy coumarin. When diluted in buffer at pH below 11 shortly before use in an enzyme assay, the substrate solution provides low background fluorescence.

Abstract: The subject invention provides a method for analyzing the metabolic activity in cells by improving the retention of a detectable reporter molecule only in intact cells where a particular enzyme is present. In particular, improved retention results from a two part process involving conjugation of haloalkyl-substituted derivatives of a reporter molecule with intracellular cysteine-containing peptides while unblocking the reporter molecule. The method for analyzing metabolic activity of cells involves the use of a substrate having the formXR-REPORTER-BLOCKwherein -BLOCK is a group selected to be removable by action of a specific analyte, to give REPORTER spectral properties different from those of the substrate,-REPORTER- is a molecule that, when no longer bound to BLOCK by a BLOCK-REPORTER bond, has spectral properities different from those of the substrate, andXR-- is a haloalkyl moiety that can covalently react with an intracellular thiol (Z--S--H) to form a thioether conjugate (Z--S--R--).

Abstract: Disclosed is a method for identifying a compound which inhibits the esterase activity of .beta.-protein. More specifically, in the method disclosed, .beta.-protein is combined with an appropriate substrate and a compound to be tested for its ability to inhibit the enzymatic activity of .beta.-protein (candidate inhibitor). Failure of .beta.-protein to act upon the substrate is an indication that the candidate inhibitor is, in fact, effective as an inhibitor of the .beta.-protein activity. Specifically, the extent to which .beta.-protein acts upon the substrate is determined and compared with the extent to which .beta.-protein acts upon the substrate in the absence of the candidate inhibitor.

Abstract: There is disclosed a composition for measurement of potassium ion concentration, which contains a) pyruvate kinase, b) glycerol kinase, c) glycerol-3-phosphate oxidase, d) peroxidase, e) a chromogen of reduced type, f) adenosine diphosphate or a salt thereof, and g) phosphoenolpyruvic acid or a salt thereof. Also disclosed is a composition for elimination of ammonium ions, which contains a) glutamate dehydrogenase, b) glucose dehydrogenase, and c) nicotinamide adenine dinucleotide of reduced type (NADH) or nicotinamide adenine dinucleotide phosphate of reduced type (NADPH).

Abstract: Methods and compositions are disclosed for determining a peroxidatively active substance (PAS). The methods comprise the step of detecting a fluorescent signal produced upon cleavage of a compound of the formula F-L-Q, wherein F is a fluorester capable of producing the signal, Q is a quencher capable of quenching the signal when linked to F, and L is a bond, or a linking group having a bond, wherein the bond is capable of being cleaved by a reaction of the PAS with a substrate of the PAS and a hydrogen donor wherein the cleavage of the bond substantially reduces the quenching. The methods have application in a wide variety of systems including assays and improved assays for analytes. Also disclosed are kits for conducting the methods and improvements in accordance with the present invention.