DE2513407A1 - Differentiation of hepato- and cardio-specific lactate dehydrogenase - in serum by selectively inhibiting the hepatospecific enzyme with heavey metal salts - Google Patents

Differentiation of hepato- and cardio-specific lactate dehydrogenase - in serum by selectively inhibiting the hepatospecific enzyme with heavey metal salts

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Publication number
DE2513407A1
DE2513407A1 DE19752513407 DE2513407A DE2513407A1 DE 2513407 A1 DE2513407 A1 DE 2513407A1 DE 19752513407 DE19752513407 DE 19752513407 DE 2513407 A DE2513407 A DE 2513407A DE 2513407 A1 DE2513407 A1 DE 2513407A1
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Germany
Prior art keywords
differentiation
ldh
lactate dehydrogenase
specific
serum
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Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
DE19752513407
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German (de)
Inventor
Karlheinz Dr Med Dr Re Schmidt
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Individual
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Individual
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Publication date
Application filed by Individual filed Critical Individual
Priority to DE19752513407 priority Critical patent/DE2513407A1/en
Publication of DE2513407A1 publication Critical patent/DE2513407A1/en
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    • 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
    • C12Q1/32—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase involving dehydrogenase

Abstract

In differentiating the hepato- and cardio-specific lactate dehydrogenase in blood serum, a heavy metal salt soln. (e.g. AgNO3 soln.) is added to the blood serum and the non-inhibited (i.e. cardiospecific) LDH is subsequently determined using a conventional procedure. Organ-specific diagnosis of diseases associated with an increase in blood LDH activity. Differentiation of cardiac and hepatic disorders is of clinical importance. The procedure permits the two LDH isoenzymes to be differentiated using conventional automatic analysers. The only step required in addition to conventional determinations of LDH is the addition of the heavy metal salt.

Description

BESCHREIBUNG DER ERFINDUNG Titel: Ein Verfahren zur Differenzierung der leber- und herzspezifischen Lactatdehydrogenase im Blutserum. DESCRIPTION OF THE INVENTION Title: A Method of Differentiation the liver- and heart-specific lactate dehydrogenase in the blood serum.

wendungs- Die Erfindung betrifft ein Verfahren zur organspezifischen biet: Differenzierung der enzymatischen Aktivität der Lactatdehydrogenase in einem kinetischen Test für medizinische Laboratorien.Application The invention relates to a method for organ-specific offer: differentiation of the enzymatic activity of lactate dehydrogenase in one kinetic test for medical laboratories.

eck: Mit derartigen Differenzierungsverfahren kann ein krankhafter 1-rozeß, der zu einem Anstieg der enzymatischen Aktivität im Serum führt, in ein bestimmtes Organ lokalisiert werden. Wichtig ist insbesondere die Unterscheidung zwischen Herz- und Lebererkrankungen wegen der klinischen Bedeutung dieser Fragestellung. Eck: With such differentiation methods, a pathological 1 process that leads to an increase in enzymatic activity in serum specific organ can be localized. The distinction is particularly important between heart and liver diseases because of the clinical importance of this issue.

stand der Es ist bekannt, daft zur Erfüllung dieser Erfordernisse Technik: eine gelelektrophoretische Trennung der organspezifischen Isoenzyme vorgenommen werden an, deren Verteilungsmuster eine Lokalisation des Kràl eits?rozesses erlaubt. Um auch im kinetischen Test eine Differenzierung zu erreichen sind vorwiegen zwei Verfahren bekannt geworden. Zum einen die Bestimmung der sog. α-Hydroxybutyratdehydrogenase, bei der die unterschiedlicne Reaktionsgeschwindigkeit der einzelnen Isoenzyme mit dem Substrat α-Hydroxybutyrat zur Differenzierung verwendet wird. Das zweite Verfahren beruht auf der unterschiedlichen Adsorption der einzelnen Isoenzyme an DEAE-Sephadex und anschließende Bestimmung der nicht a dsorbierten LDH-Aktivität.It is well known that these requirements have been met Technique: a gel electrophoretic separation of the organ-specific isoenzymes carried out whose distribution pattern allows localization of the cranial process. In order to achieve a differentiation in the kinetic test, two are predominant Procedure became known. On the one hand, the determination of the so-called α-hydroxybutyrate dehydrogenase, in which the different reaction rates of the individual isoenzymes with the substrate α-hydroxybutyrate is used for differentiation. The second Procedure is based on the different adsorption of each Isoenzymes on DEAE-Sephadex and subsequent determination of the non-adsorbed LDH activity.

ritik des Die gelelektrophoretische Auftrennung ermöglicht eine Standes der rechnik: optimale Organdifferenzierung ist aber für die analytisch Routine in Klinik und Praxis zu aufwendig. Die kinetische Tests mit Variation des Substrats oder Acisorption an unlösliche Träger sind einerseits wenig spezifisch, anderer seits erfordern sie zusätzliche Arbeitsgänge und sind nur teilweise automatisierbar.ritic of The gel electrophoretic separation enables a stand der arithmetic: optimal organ differentiation is in for the analytical routine Clinic and practice too expensive. The kinetic tests with variation of the substrate or absorption to insoluble carriers are on the one hand not very specific, on the other hand on the other hand, they require additional work steps and can only be partially automated.

Aufgabe: Der Erfindung liegt die Aufgabe zugrunde, eine spezifisch Trennung der der herz- bzw. leber typischen LDH zu erreicht und die quantitative Bestimmung ohne zusätzliche Arbeitsg, auf handelsüblichen Enzymautomaten durchzuführen.Task: The invention is based on the task of providing a specific Separation of the heart and liver typical LDH and the quantitative Determination without additional work to be carried out on commercially available enzyme machines.

Lösung: Diese Aufgabe wird erfindungsmäßig dadurch gelöst, daß zu dem LDH-haltigen Blutserum eine Schwermetallsalzlösung zu gesetzt wird, die hochselektiv eine Hemmung der leberspez: fischen LDH herbeiführt. Danach kann die ungehemmte d.h.Solution: According to the invention, this object is achieved in that to A heavy metal salt solution is added to the LDH-containing blood serum, which is highly selective an inhibition of the liver-specific: fish LDH brings about. After that, the uninhibited i.e.

herzspezifische LDH ohne jede verfahrenstechnische derui auf demselben Gerät bestimmt werden, das auch zur Bestimmt der Gesamt-LDH Verwendung findet. heart-specific LDH without any procedural derui on the same Device can be determined which is also used to determine the total LDH.

Erzielbare Die mit der Erfindung erzielten Vorteile bestehen darin, Rtorteile: daß ohne jeden zusätzlichen apparativen Aufwand eine orga spezifische Differenzierung der Lactatdehydrogenase ermöglicht wird. Mit Ausnahme des Schwermetallsalzes werden keine Chemikalien benötigt, zusätzliche Arbeitsgänge wie Zentrifugieren, Inkubieren sind nicht erforderlich.Achievable The advantages achieved with the invention are Rtorteile: that an orga specific Differentiation of lactate dehydrogenase is made possible. With the exception of the heavy metal salt no chemicals are required, additional operations such as centrifugation, Incubation is not required.

ihrungsbeispiel Photometrischer Test im Ultraviolett Wellenlänge 334 nm Schichtdicke der Küvette 1 cm Temperatur 25°C Folgende Volumina dosieren: Serum 4 Vol AgNO3 1 mM 1 Vol Bestimmungsansatz: 50 MM Phosphatpuffer pH 7,5 500 jil o,18 mM NADEL Serum + AgNO3-Lösung 10 µl o,6 mM Pyruvat 20 Berechnung der Vol.Aktivität nach Vol.Akt. = 441 ' tan n (mU/ml) Anschließend Berücksichtigung der 20 %igen Verdünnung durch die AgNO3-Lösung.Evaluation example Photometric test in the ultraviolet wavelength 334 nm layer thickness of the cuvette 1 cm temperature 25 ° C dose the following volumes: Serum 4 vol AgNO3 1 mM 1 vol determination mixture: 50 MM phosphate buffer pH 7.5 500 jil 0.18 mM NADEL serum + AgNO3 solution 10 µl o, 6 mM pyruvate 20 Calculation of the activity by volume according to vol. = 441 tan n (mU / ml) Then take into account the 20% dilution through the AgNO3 solution.

Die Ergebnisse zahlreicher Bestimmungen mit statistischer Auswertung sind in der Abbildung angegeben. The results of numerous determinations with statistical evaluation are indicated in the figure.

Claims (1)

PATENTANS PRÜCHE PATENTAN'S CLAIMS Oberbegriff: Ein Verfahren zur Differenzierung der leber- und herzspezifischen Lactatdehydrogenase (LDH) im Blutserum Kennzeichnen- dadurch gekennzeichnet, daß zu dem Blutserum eine der Teil: Schwermetallsalzlösung zugesetzt und anschließend die nicht gehemmte d.h. herzspezifische LDH in einem üblichen Verfahren bestimmt wird.Generic term: A method for differentiating the liver and heart-specific Characteristic of lactate dehydrogenase (LDH) in blood serum - characterized in that to the blood serum one of the part: heavy metal salt solution added and then the non-inhibited, i.e. heart-specific, LDH is determined in a conventional method will. ERFINDERBENENNUNG Weitere Personen sind meines Wissens an der Erfindung nicht beteiligt. Der Titel der Erfindung lautet: "Ein Verfahren zur Differenzierung der leber-und herzspezifi schen Lactatdehydrogenase im Blutserum". INVENTOR NAME Other people are involved in the invention as far as I know not involved. The title of the invention is: "A method of differentiation of the liver- and heart-specific lactate dehydrogenase in the blood serum ".
DE19752513407 1975-03-26 1975-03-26 Differentiation of hepato- and cardio-specific lactate dehydrogenase - in serum by selectively inhibiting the hepatospecific enzyme with heavey metal salts Pending DE2513407A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
DE19752513407 DE2513407A1 (en) 1975-03-26 1975-03-26 Differentiation of hepato- and cardio-specific lactate dehydrogenase - in serum by selectively inhibiting the hepatospecific enzyme with heavey metal salts

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
DE19752513407 DE2513407A1 (en) 1975-03-26 1975-03-26 Differentiation of hepato- and cardio-specific lactate dehydrogenase - in serum by selectively inhibiting the hepatospecific enzyme with heavey metal salts

Publications (1)

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DE2513407A1 true DE2513407A1 (en) 1976-10-14

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2830453A1 (en) * 1977-07-11 1979-02-01 Eastman Kodak Co METHOD FOR DETERMINING THE ACTIVITY OF ISOENZYME
EP0292838A1 (en) * 1987-05-28 1988-11-30 Abbott Laboratories A method and reagent for determining LD-1 isoenzyme
EP0352547A2 (en) * 1988-07-15 1990-01-31 International Reagents Corporation LDH1 assay

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2830453A1 (en) * 1977-07-11 1979-02-01 Eastman Kodak Co METHOD FOR DETERMINING THE ACTIVITY OF ISOENZYME
EP0292838A1 (en) * 1987-05-28 1988-11-30 Abbott Laboratories A method and reagent for determining LD-1 isoenzyme
EP0352547A2 (en) * 1988-07-15 1990-01-31 International Reagents Corporation LDH1 assay
EP0352547A3 (en) * 1988-07-15 1991-07-31 International Reagents Corporation Ldh1 assay

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