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
  • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
  • C12Q1/44—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving esterase
  • C12Q1/46—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving esterase involving cholinesterase
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
  • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
  • G01N33/525—Multi-layer analytical elements

Definitions

• the present invention relates to a detector kit for the indication of nerve gases and other cholinesterase-inhibitors.
• the present invention relates to a detector kit in one part, which is extremely simple to use and which can be produced at a Low cost and with simple means.
• the invention is primarily meant to be used in a war situation where nerve gases might be used.
• the person, who is going to use the equip ⁇ ment may in many cases be under a very strong physical and psychical pressure, the light conditions may be very poor and moreover, this person is wearing a protective mask and a protective suit which among other things involves the use of thick and clumsy gloves.
• a protective mask and a protective suit which among other things involves the use of thick and clumsy gloves.
• the equipment should be of an easily manageable size and be so devised that the indication can be performed with simple movements of one's hand. It is consequently disadvantageous with an equipment which consists of several small parts that are to be moved about during the indication. Moreover, the equipment should be so constructed that one can easily bring the enzyme and the substrate in contact with each other" during the indication.
• the detector kit can in a folded form keep the substrate and the enzyme away from the ambient air without using a separate outer wrapping.
• the enzyme must be wetted for the indication.
• the enzyme surface has been wetted by means of especially provided water before the indication.
• the detector kit is therefore near the enzyme provided with a container comprising water to wet the choLinesterase.
• the container can be made to burst by means of the pressure from a finger.
• the container may comprise a freezing point depressing means, for instance glycol, in order to make the detector kit usable in the cold. It is of very great practical importance that the individual user does not have to start looking for water when he wants to use the detector kit.
• liquid container is protected from un ⁇ intentional puncturing during storage and transport. If the container has been punctured, this must be recognizable when the wrapping is broken. It is here of great importance if one can, not only visually, verify that the enzyme has been wetted, as this may be difficult during troublesome conditions.
• the liquid container is so placed on the detector kit that it is broken and becomes loose ;after being punctured and thus clearly shows in this way that it has been punctured.
• fig 1a shows a basic embodiment of the invention
• fig 1b is a sectional view taken on the Line A-A of fig 1a
• fig 2 shows an embodiment of the invention in a Longitudinal sectio ⁇ nal view with a liquid container close to the enzyme
• ig 3a shows another embodiment of the invention in a longitudinal sectional view with a liquid container close to the enzyme
• ig 3b shows the invention of fig 3a in a cross-sectional view through the enzyme section
• ig 4 shows an embodiment of the invention with a three-section detec ⁇ tor kit, which can be folded twice and is provided with a liquid container
• fig 5a shows an embodiment of the invention which is airtight per se during storage
• fig 5b is a sectional view taken on the line B-B of fig 5a with the detector kit folded and provided with a desiccant and a sepa ⁇ rating foil
• fig 6a shows still
• choLinesterase 2 On a support 1 there are placed the enzyme choLinesterase 2 and also a substrate 3 for the enzyme.
• the choLinesterase can suitably be immobi ⁇ lized and stabilized on paper.
• the enzyme papers prepared in this way are dried, first in ambient air and then in an exsiccator under vacuum.
• the substrate which has proved to have the best properties for an en ⁇ zymatic colour reaction, is 2,6-dichloro indophenyl acetate (DCIPA).
• DCIPA 2,6-dichloro indophenyl acetate
• the working up has been modified in as much, as DCIPA has been precipitated in water and then recrystallized in n-hexane:diethyl ether.
• the substrate paper is produced by dissolving DCIPA in a suitable soL- vent, e.g. acetone, carbon tetrachloride, chloroform, dichloromethane or 1,2-dichloroethane, and then applying the solution in drops onto the paper. So much of the.solution is used that the whole paper is evenly.
• a suitable soL- vent e.g. acetone, carbon tetrachloride, chloroform, dichloromethane or 1,2-dichloroethane
• the surface concentration i.e. the amount of DCIPA per cm , is varied by changing the concentration of DCIPA in the solution.
• the surface concentration is not critical as long as a sufficiently Large amount of the substrate is available in order to get a clear change in the enzymatic colour reaction. Be ⁇ cause of the losses that one must take into account during storage, in the form of decomposition and possibly sublimation of the substrate,
• the Latter test has been accomplished by placing paper, impregnated with DCIPA, between two pieces of the plastic in question. The whole package has then been stored in heat at + 60 C together with a de- siccant. After 46 days the sample was taken out and examined with re ⁇ gard to whether DCIPA had diffused into the plastic or not and with regard to how well the DCIPA-papers functioned during indication.
• plastic materials are considered usable for the pro- duction of detector kits using enzyme: Plexiglas,. polystyrene and polycarbonate.
• a liquid container 4 can be placed close to the enzyme.
• the container is provided with a pin 5, which perforates the support 1 when a finger is pressed there- on ; so that water will pass to the enzyme. It is also possible with other ways of arranging a Liquid container close to the enzyme.
• figs 3a and 3b is illustrated how the water container 4 can be placed on the support 1 by means of a casing 10 pressed on to the support.
• the casing is hard and brittle and is cracked on use, where ⁇ after it loosens from the support 1 thus giving a clear indication of the fact that the enzyme has been wetted. (Important for instance if wetting should occur unintentionally during storage.)
• the sharp cracking sound and the detachment from the support give clear signals during correct handling that the enzyme has become satisfac ⁇ tory wetted.
• fig 4 is illustrated a detector kit in three sections with the en ⁇ zyme 2 on the middle section.
• a liquid container 4 is placed on a second section of the detector kit.
• the enzyme 2 which can be provided with pins 5
• the container 4 can be made to break so that water will wet the enzyme.
• the substrate 3 is placed on the third section of the detector kit. After the wetting of the enzyme the second section is unfolded, and the third section is folded over the enzyme so that the enzyme and the substrate are brought into contact with each other.
• figs 5a and 5b is illustrated an embodiment of the invention with a sealing device 6 around the substrate and the enzyme.
• the sealing device consists of raised edges arranged on the support and extending aroung the enzyme and the substrate and of such dimensions that, when the detector kit is folded, one of the raised edges will come into a firm engagement with the other raised edge.
• the two edges can have such a cross-section that they will snap into each other. In many cases, however, a sufficient sealing is obtained already with straight edges.
• the raised edges are circular but other forms can of course be used.
• figs 6a and 6b is illustrated another embodiment, which also is airtight per se during storage.
• the edges 7 of the support for the enzyme and the substrate are here welded together or sealingly faste ⁇ ned to each other in another way, so that a folded detector kit is being formed comprising an airtight space in which the substrate and the enzyme are located. In use, the detector kit is torn apart, and then the procedure is continued as described above.
• the enzyme and the substrate are kept apart during storage by means of. a foil 8 over the substrate.
• the foil can be torn away after the detector kit has been opened or be so fastened over the' substrate that it is automatically torn away when the detector kit.is opened.
• a thin aluminium foil has been used with a good result.

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• Life Sciences & Earth Sciences (AREA)
• Health & Medical Sciences (AREA)
• Chemical & Material Sciences (AREA)
• Engineering & Computer Science (AREA)
• Immunology (AREA)
• Molecular Biology (AREA)
• Hematology (AREA)
• Organic Chemistry (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Zoology (AREA)
• Urology & Nephrology (AREA)
• Microbiology (AREA)
• Wood Science & Technology (AREA)
• Biomedical Technology (AREA)
• Physics & Mathematics (AREA)
• Analytical Chemistry (AREA)
• Biochemistry (AREA)
• General Health & Medical Sciences (AREA)
• Biotechnology (AREA)
• Pathology (AREA)
• Biophysics (AREA)
• General Physics & Mathematics (AREA)
• Medicinal Chemistry (AREA)
• Food Science & Technology (AREA)
• Cell Biology (AREA)
• Bioinformatics & Cheminformatics (AREA)
• General Engineering & Computer Science (AREA)
• Genetics & Genomics (AREA)
• Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
• Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
• Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)

Applications Claiming Priority (2)

Publications (1)

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Cited By (6)

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Citations (4)

• 1984
  • 1984-04-02 SE SE8401798A patent/SE8401798L/ unknown
• 1985
  • 1985-04-02 EP EP19850902181 patent/EP0177600A1/en not_active Ceased
  • 1985-04-02 WO PCT/SE1985/000158 patent/WO1985004424A1/en not_active Application Discontinuation
  • 1985-04-02 AU AU42307/85A patent/AU4230785A/en not_active Abandoned
  • 1985-04-02 JP JP50178985A patent/JPS61501679A/ja active Pending

Patent Citations (4)

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