Boehringer Mannheim Gmbh

Joachim Siedel

Joachim Ziegenhorn

Albert Roder

Hans Mollering

Bernd Vogt

Brigitte Pautz

Lieselotte Schellong

Hans-Georg Batz

Helmut Lenz

Winfried Albert

Hans Seidel

Helmgard Gauhl

Michael-Harold Town

Johnny Staepels

Gerd Zimmermann

Rolf Deeg

Ulrich Neumann

Gunter Frey

Ulrich Mayr

Rupert Herrmann

Christian Birkner

August W Wahlefeld

Helmut Schlumberger

Uwe Herrmann

Herbert Buschek

Lorenz Kerscher

Cornelia Kruse-Muller

Michael Harold Town

Hans-George Batz

Liselotte Schellong

Blood substitute suppression by peroxides

The present invention is concerned with the use of certain 3-aminopyrazolo-heterocyclic compounds for the colorimetric determination of hydrogen peroxide, hydrogen peroxide-forming systems, peroxidase, peroxidate-active substances and electron-rich aromatic compounds. The present invention is also concerned with corresponding processes of determination and with agents appropriate therefor. Furthermore, the present invention is concerned with new 3-aminopyrazolo-heterocyclic compounds.

3-aminopyrazolo heterocyclic derivatives, and use for colorimetric

The present invention provides a process for the specific determination of the serum fructosamine content in blood or samples derived from blood by reaction with an appropriate color reagent and measurement of the color change thereby brought about, wherein, before the color reaction, non-specific reducing-acting and/or turbidity-causing sample components are removed at approximately neutral pH value, subsequently the pH is adjusted to a value of from 10 to 12 and the color reagent is added thereto. The present invention also provides a reagent mixture for the specific determination of the serum fructosamine content in blood or samples derived from blood, wherein it comprises a reagent for the removal of non-specific reducing-acting and/or turbidity-causing sample components, a rebuffering reagent with a buffer which has a pH value in the range of from 10.5 to 12.5 and a color reagent for the detection of fructosamine.

Reagent for specific determination of fructosamine

A process for the determination of substrate or enzyme activities by the use of a redox reaction as a measurement reaction is carried out in the presence of one or more additionally added tetrazolium salts to remove disturbing substances. The tetrazolium salts have the formula ##STR1## in which R.sup.1 is a hydrogen atom, a carboxyl group or an alkyl, phenyl, nitrophenyl, dinitrophenyl, carboxyl-substituted phenyl or trialkylammoniumphenyl radical, R.sup.2 is a phenyl, nitrophenyl, biphenylyl or naphthyl radical, R.sup.3 is a phenyl, carboxyl-substituted phenyl, carboxyl-substituted hydroxyphenyl or dimethylthiazolyl radical, and A.sup..crclbar. is a monovalent anion. The formazanes formed by reaction with reducing substances do not absorb light at all, or absorb light only to a negligible extent, at the measurement wavelength of the redox reaction. New compounds included within the structural formula are those in which R.sup.1 is carboxy, phenyl, 2-carboxyphenyl, 4-carboxyphenyl, 2,4-dinitrophenyl or 4-trimethylammoniumphenyl, R.sup.2 is phenyl, and R.sup.3 is phenyl or 2-carboxyphenyl; with the proviso that, when R.sup.3 is unsubstituted phenyl or 2-carboxyphenyl, R.sup.1 may not be unsubstituted phenyl.

Process and reagent for the determination of substrates or enzyme

This invention teaches a process for reducing protein matrix effects in assays for serum fructosamine. Blood or blood derived samples are used, and one adds two reagents, one of which reduces interference caused by non-specific reducing substances, the other of which eliminates turbidity. Incubation follows, and then the pH of the sample is adjusted and color forming reagent is added. In one embodiment, the incubation time is only 1-15 minutes. In another embodiment, the first reagent contains peroxidase.

Process for reduction of the matrix effect in a fructosamine

Process for the specific determination of the serum fructosamine content in blood or in samples obtained from blood by reaction with an appropriate color reagent and measurement of the color change thereby brought about, in which before the color reaction sample components with a non-specific reducing action and/or causing turbidity are removed and subsequently the color reagent is added at a pH value of from 10 to 12. The sample components are removed by treatment at approximately neutral pH value with a reagent composition comprising at least one enzymatic oxidation agent, optionally together with peroxidase and/or catalase and/or lipase, as well as with at least one SH group-blocking substance. A kit for the specific determination of the serum fructosamine content in blood or samples obtained from blood, comprises said reagent composition, a rebuffering reagent with a buffer which has an alkaline pH value and a color reagent for the detection of fructosamine.

Process and reagent for the specific determination of fructosamine

The present invention provides a process for the improved colorimetric determination of hydrogen peroxide as formed by a hydrogen peroxide-producing oxidase, by addition of a chromogenic system and measurement of the colored material formed, wherein superoxide dismutase (E.C. 1.15.1.1) is added to the reagent solution. The present invention also provides a reagent for the improved colorimetric determination of hydrogen peroxide, comprising a hydrogen peroxide-producing oxidase, a chromogenic system, a buffer and optionally adjuvant enzymes, wherein it also contains superoxide dismutase.

Process and reagent for the improved quantitative colorimetic

A process for the determination of fructosamine in body fluids by the reaction of a sample solution with a color reagent, wherein the sample liquid is mixed with a buffer solution having a pH value of from 9 to 12, a color-forming reagent and uricase, as well as with at least one detergent, and the chronological change of the extinction is measured kinetically in a temperature range of from 20.degree. to 40.degree. C. at the earliest after 5 minutes.

Process and reagent composition for determination of fructosamine in

A process for the determination of substrate or enzyme activities by the use of a redox reaction as a measurement reaction is carried out in the presence of one or more additionally added tetrazolium salts to remove disturbing substances. The tetrazolium salts have the formula ##STR1## in which R.sup.1 is a hydrogen atom, a carboxyl group or an alkyl, phenyl, nitrophenyl, dinitrophenyl, carboxyl-substituted phenyl or trialkylammoniumphenyl radical, R.sup.2 is a phenyl, nitrophenyl, biphenylyl or naphthyl radical, R.sup.3 is a phenyl, carboxyl-substituted phenyl, carboxyl-substituted hydroxyphenyl or dimethylthiazolyl radical, and A.sup..crclbar. is a monovalent anion. The formazanes formed by reaction with reducing substances do not absorb light at all, or absorb light only to a negligible extent, at the measurement wavelength of the redox reaction. New compounds included within the formula are those in which R.sup.1 is carboxy, phenyl, 2-carboxy, phenyl, 2-carboxyphenyl, 4-carboxyphenyl, 2,4-dinitrophenyl or 4-trimethylammoniumphenyl, R.sup.2 is phenyl, and R.sup.3 is phenyl or 2-carboxyphenyl; with the proviso that, when R.sup.3 is unsubstituted phenyl or 2-carboxyphenyl, R.sup.1 may not be unsubstituted phenyl.

Removal of interfering reducing substances with tetrazolium salts in

The present invention provides a process for the determination of iron in body fluids by liberation of the bound iron, reduction to Fe.sup.2+, addition of a color system appropriate for the detection of iron and photometric measurement in a tenside-containing sample solution, wherein a fatty acid polyethylene glycol ester, an alkanol polyglycol ether and at least 1 mole/liter quanidine hydrochloride are added to the sample solution. The present invention also provides a reagent for the determination of iron in serum, containing a reducing agent, a color material system appropriate for the detection of iron and guanidine hydrochloride and at least one tenside, wherein it contains a fatty acid polyethylene glycol ester and a alkanol polyglycol ether.

Method for determining iron in a body fluid sample

The present invention provides a process for the determination of total bilirubin in samples of body fluids. The method comprises the incubation of a sample of body fluid with both subtilisin which completely liberates bilirubin from its albumin and with bilirubin oxidase as a measurement of total bilirubin content in said body fluid sample.

Process and reagent for the determination of total bilirubin

The present invention provides a process for the specific determination of HDL cholesterol in serum or plasma by incubation with a cholesterol detection system, containing cholesterol oxidase and cholesterol esterase in a buffered aqueous medium, and measurement of a product of the cholesterol oxidase reaction or of the oxygen consumption, wherein a sample to be tested is incubated in the presence of a salt of a bile acid or of a bile acid derivative or of dioctylsulphosuccinate, a first measurement is then carried out, subsequently a non-ionic, polyethylene oxide group-containing detergent or a secondary alkane sulphonate is added, again incubated and a second measurement is carried out, the amount of HDL cholesterol being determined from the difference between the first and second measurement. The present invention also provides a reagent for the specific determination of HDL cholesterol in serum or plasma containing cholesterol oxidase, buffer and cholesterol esterase, wherein, referred to the final solution, it contains

Process and reagent for the specific determination of HDL cholesterol in

The present invention provides a method of use for a hydrogen peroxide-forming oxidase, wherein the enzyme is obtained from Streptomycetaceae and at 25.degree. C., in 0.15 mol/liter potassium phosphate (pH 7.9), in the presence of surface-active substances, still shows after 2 days an activity of at least 40% of the initial activity. This enzyme is useful in the determination of sarcosine, creatine and creatinine.

Method for the determination of sarcosine creatine or creatinine

The present invention provides 1-methylhydantoinase, which hydrolyses 1-methylhydantoin in the presence of a nucleoside triphosphate and of polyvalent metal ions. The present invention also provides a process for obtaining 1-methylhydantoinase and a reagent containing it. Furthermore, the present invention provides a process for the determination of creatinine by the conversion of the creatinine with creatinine deiminase (E.C. 3.5.4.21) into 1-methylhydantoin, hydrolysis of the latter with the 1-methylhydantoinase in the presence of nucleoside triphosphate and of polyvalent metal ions and determination

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Bernried, Germany

Joachim Siedel