Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
  • C12Q1/26—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
  • C12Q1/28—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase involving peroxidase
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q2326/00—Chromogens for determinations of oxidoreductase enzymes
• • 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
  • Y10S435/00—Chemistry: molecular biology and microbiology
  • Y10S435/805—Test papers
• • 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
  • Y10S436/904—Oxidation - reduction indicators

Definitions

• ABSTRACT Test strips for the detection of peroxidatively active substances in body fluids, e.g., for the detection of blood in urine, comprising a carrier containing a. a hydroperoxide;
• R is pyridyl or phenyl, optionally substituted by lower alkyl or alkoxy, and R is hydrogen or lower alkyl.
• the present invention relates to a test strip for the de tection of very small amounts of blood and of other peroxidatively-active substances in the body fluids.
• hemorrhages are caused, for example, by tumors, ulcers or inflammations in the organs in question.
• free hemoglobin can also occur in the urine and plasma.
• Blood and hemoglobin are .peroxidatively-active, i.e., they liberate oxygen from hydroperoxides and transfer it to certain acceptors.
• Myoglobin which is also peroxidatively-active, is found, for example, in the urine after a cardiac infarct.
• blood occurs especially frequently in the urine when calculi are present in the bladder or kidneys.
• Rapid test compostions are absorbent carriers, usually papers, which are impregnated with all the reagents needed for the detection reaction and which, after simply dipping into the body fluid to be tested, show a color reaction. Because of the great importance which these rapid tests have recently achieved, they have also been developed in various ways for the detection of blood in body fluids.
• German Pat. Spec. No. 1,242,905 describes test papers which, as activating additives, contain certain quinoline derivatives, preferably quinine. It is certainly asserted that the sensitivity can be considerably increased by means of these additives but, according to our own investigations, it is practically impossible, with the so improved tests strips, to detect, for example, blood in a concentration of 5 erythrocytes /mm", which would be necessary for a differentiation between normal and pathological values.
• test papers for the detection of peroxidatively-active substances with a previously unachievably high sensitivity are obtained by utilizing certain vinylpyridine compounds as activators.
• the vinyl pyridine compounds showing this efficacious action are of the formula wherein R is pyridyl or phenyl, optionally substituted by lower alkyl or alkoxy, and
• R is hydrogen or lower alkyl.
• Preferred compounds of general formula (I) are those in which R, is hydrogen.
• the lower alkyl and alkoxy radicals can contain up to 4 carbon atoms, the methyl and methoxy radicals being preferred.
• a test strip for the detection of peroxidativelyactive substances in body fluids comprising a carrier containing a hydroperoxide, at least one chromogen and, as activator, at least one compound of general formula (I).
• the sensitivity of the detection reaction is increased by some of the compounds of general formula (I) to such an extent that it is even possible to detect individual erythrocytes in urine, which are visible on the test paper as colored dots.
• a-styryl-pyridine it is possible to produce a test paper with which it is possible to detect 5 erythrocytes/mm urine. This corresponds to a blood dilution of 1110.
• the activating agent used according to the present invention can be employed in amounts of about 0.05 1.0g, preferably of 0.2 0.5g, per ml. ofimpregnation solution.
• an organic hydroperoxide an oxidation indicator (chromogen), a buffer and a surface-active agent, as well as, if desired, a phosphoramide, for example phosphoric acid trimorpholide, for stabilization, as well as conventional adjuvants.
• hydroperoxides there can be used, for example, diisopropyl-benzene hydroperoxide or 2,5-dimethylhexane-2,S-dihydroperoxide, and as indictors, there can be used, for example, those of the benzidine series, such as o-tolidine, or those of the heterocylic azine series, such as bis-(N-ethyl-quinol-Z-one)-azine or (N- methyl-benz-thiazol-2-one)-(1-ethyl-3-phenyl-5- methyltriazol-Z-one)-azine (see German Pat.Spec No. 1,648,840).
• benzidine series such as o-tolidine
• heterocylic azine series such as bis-(N-ethyl-quinol-Z-one)-azine or (N- methyl-benz-thiazol-2-one)-(1-ethyl-3-phenyl-5
• the indicator can be used in amounts of from 0.05 g., preferably of 0.2 1.0 g., per 100 ml. impregnation solution.
• buffer there can be used, for example, a citrate. phosphate, phthalate or succinate buffer. the pH value and capacity being so chosen that. after dipping the test strip into a body fluid, a pH value of 4 7. preferably 5 3 6, is obtained thereon.
• a complex-forming agent for example, sodium metaphosphate or ,ethylenediamine-tetraacetic acid, in order to avoid falsely positive reactions which could be caused bytraces of metals.
• test papers can, because of the relatively large amounts 7 of water-soluble substances present, tend to bleed, it is practical to add to the formulation a thickening agent, for example, methyl cellulose and, in particular, polyvinylpyrrolidone, preferably in amounts of about 0.5 5 g. per 100 ml. g I
• a thickening agent for example, methyl cellulose and, in particular, polyvinylpyrrolidone, preferably in amounts of about 0.5 5 g. per 100 ml. g I
• wetting agent there is preferably used a longchainedorganic sulphate or sulphonate, for example sodium 'dodecyl-benzenesulphonate, dioctyl sodium sulphosuccinate or sodium lauryl sulphate, which, as is known, stabilizes radical cations, such as oxidized otolidine.
• the wetting agent can be added to the impregnation solutionin amounts of from 0.5 5%, preferably of'l 3%.
• absorbentcarriers for example, filter paper, cellulose or synthetic fibre fleeces
• solutions of the reagents'in readily volatile liquids This is preferably carried out in two or three separate steps.
• impregnation is first carried out with a solution which contains a hydroperoxide, wetting agent, buffer and optionally a thickening agent.
• impregnation is carried out with a solution of an indicator and of an activator of general formula (1).
• test strips according to the present invention are cut up into strips and preferably sealed between a synthetic resin film and a fine-mesh material in the manner described in German Pat. Spec. No. 2,118,455.
• Filter paper is successively impregnated with the following solutions and dried'at' 40C.:
• A'white test paper is obtained which, upon dipping into blood-containing urine, becomes blue-green colored after about 5 20 seconds. If the erythrocytes are intact, then the papers are blue-green speckled. If hemolysis has taken place or if free hemoglobin was initially present in the urine, then the papers are uniformly blue-green colored.
• the sensitivity is about 5 erythrocytes/mm or the corresponding amount of hemoglobin. A lower number of intact'er'ythrocytes can, under certain circumstances, still bring about'individual green dots on the test paper. The sensitivity with regard to myoglobin corresponds to that for hemoglobin.
• Filter paper is impregnated with the followingsolutions and dried at 40C.:
• Test papers are obtained with'practically the same properties as those described in Example I.
• the sensitivety is between about 5 and 20 erythrocytes/mm .'de-
• Test strip composition for the detection of peroxidatively-active substances in body fluids, comprising a carrier containing a hydroperoxide, at least one chromogen and, as an activator, a compound of the formula:
• R is pyridyl or a phenyl optionally substituted by lower alkyl or alkoxy;
• R is hydrogen or lower alkyl.
• Test strip as claimed in claim 1 wherein R in the formula of said activator compound is pyridyl substituted by methyl or methoxy.
• Test strip as claimed in claim 1 wherein R in the formula of said activator compound is phenyl substituted by methyl or methoxy.
• Test strip as claimed in claim 1 wherein R in the formula of said activator compound is hydrogen.
• test strip as claimed in claim 1 wherein said activator compound is Z-(phenylvinyl)-4-methylpyridine.
• Test strip as claimed in claim 1 wherein said activator compound is pyridyl-2-pyridyl-4-ethylene.
• Test strip as claimed in claim 1 wherein said activator compound is 2-(4-methylphenylvinyl)-pyridine.
• Test strip as claimed in claim 1 wherein said activator compound is 2-(4-methoxyphenylvinyl)-pyridine.
• test strip as claimed in claim 1 wherein said activator compound is di-(pyridyl-2)-ethylene.
• Test strip as claimed in claim 1 wherein said hydroperoxide is diisopropyl-benzene hydroperoxide or 2,5dimethylhexane-2,S-dihydroperoxide.
• Test strip as claimed in claim 1 comprising citrate buffer ethylenediaminetetraacetic: acid, disodium salt dodecyl-benzene-sulfonic acid sodium salt 2,S-dimethylhexane-Z.S-dihydroperoxide phosphoric acid trimorpholide o-tolidine o-stilbazole.
• Method of detecting a peroxidatively active substance in a body fluid comprises contacting a sample of said body fluid with a test strip composition as claimed in claim 1 and observing a color change in said test strip composition as an indication of the absence or presence of the peroxidatively active sub stance.

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• Chemical & Material Sciences (AREA)
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• Proteomics, Peptides & Aminoacids (AREA)
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• General Engineering & Computer Science (AREA)
• General Health & Medical Sciences (AREA)
• Genetics & Genomics (AREA)
• Investigating Or Analysing Biological Materials (AREA)
• Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
• Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)

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Cited By (42)

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Citations (5)

• 1974
  • 1974-01-01 AR AR256902A patent/AR204182A1/es active
  • 1974-11-25 US US526661A patent/US3917452A/en not_active Expired - Lifetime
  • 1974-12-11 CA CA215,794A patent/CA1027832A/fr not_active Expired
  • 1974-12-13 FI FI3610/74A patent/FI57025C/fi active
  • 1974-12-13 ZA ZA00747948A patent/ZA747948B/xx unknown
  • 1974-12-14 DD DD183041A patent/DD117530A5/xx unknown
  • 1974-12-16 CH CH1668074A patent/CH595635A5/xx not_active IP Right Cessation
  • 1974-12-16 SE SE7415757A patent/SE399129B/xx unknown
  • 1974-12-16 GB GB5417074A patent/GB1443133A/en not_active Expired
  • 1974-12-16 IT IT30609/74A patent/IT1027722B/it active
  • 1974-12-16 JP JP14434274A patent/JPS5412360B2/ja not_active Expired
  • 1974-12-18 NL NL7416484.A patent/NL164965C/xx not_active IP Right Cessation
  • 1974-12-19 HU HU74BO00001527A patent/HU172449B/hu unknown
  • 1974-12-19 FR FR7441994A patent/FR2283441A1/fr active Granted
  • 1974-12-19 AT AT1014474A patent/AT337909B/de not_active IP Right Cessation

Patent Citations (5)

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