SU976958A1 - Method of diagnosis of myocardial infarction - Google Patents

Method of diagnosis of myocardial infarction Download PDF

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Publication number
SU976958A1
SU976958A1 SU741999385A SU1999385A SU976958A1 SU 976958 A1 SU976958 A1 SU 976958A1 SU 741999385 A SU741999385 A SU 741999385A SU 1999385 A SU1999385 A SU 1999385A SU 976958 A1 SU976958 A1 SU 976958A1
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USSR - Soviet Union
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mercaptoethanol
glycogen
sodium
solution containing
body fluid
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SU741999385A
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Russian (ru)
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Краузе Эрнст-Георг
Вилль Хорст
Бем Манфред
Волленберген Альберт
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Феб Арцнаймиттельверк Дрезден (Инопредприятие)
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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
    • C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
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  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

1451004 Isophosphorylase detection ARZNEIMITTELWERK DRESDEN VEB 29 Jan 1974 [7 Feb 1973] 04061/74 Heading GIB A process for the determination of the activity of total phosphorylase or of individual isophosphorylases in body fluids, particularly in blood, comprises contacting concentrated body fluid (or a body fluid pre-treated with starch) with a glycogen-containing substrate for phosphorylases, the enzyme activity being determined by determining the inorganic phosphates or amylose-like side chains of glycogen formed. The process may comprise mixing a concentrated body fluid with a solution containing glycylglycine, mercaptoethanol and ethylene diamine tetra acetic acid in a 1:1 ratio or body fluid pre-treated on a starch column by adsorption and washing with a solution of sodium fluoride, sodium #-glycerophosphate and mercaptoethanol (pH 6À1, 4‹C), followed by elution of the phosphorylase at 30 C and pH 6À8 with a solution of sodium #-glycerophosphate, mercaptoethanol and ethylene diamine-tetra acetic acid, and mixing the resulting fluid in a 5:3 ratio with a glycogen-containing solution (the substrate), then adding 2 parts of a solution containing glucose-1-phosphate and adenosine monophosphate, the mixture being incubated for 60 minutes, thereafter denaturing with trichloroacetic acid, the phosphate content being subsequently determined. An alternative process comprises pretreating a body fluid on a starch column as above, diluting the eluate with 1À2M saccharose in a 2:1 ratio, applying the diluted eluate to an acrylamide separation gel containing 0À1% glycogen (the substrate) and buffered with a solution containing tris-(hydroxymethyl)-amino methane, ethylenediamine-tetra acetic acid and borric acid, separating by disc electro-phoreses and subsequently, after incubation in an aqueous solution containing glucose-1-phosphate, adenosine monophosphate, sodium #-glycerophosphate, ethylene diaminetetra acetic acid, mercaptoethanol and sodium fluoride, the formation of the amylose-like side chains on the glycogen being made visible by iodine-potassium iodide coloration.

Description

Изобретение относитс  к медицине , а именно к диагностике инфаркта миокарда.This invention relates to medicine, in particular to the diagnosis of myocardial infarction.

Известен способ диагностики инфаркта миокарда путем определени  активности фермента в сыворотке крови tl.A known method for the diagnosis of myocardial infarction by determining the enzyme activity in the serum tl.

Однако известный способ  вл етс  трудоемким, так как в сыворотке находитс  лишь незначительное количество фосфорилазы при большом избытке других протеинов, что мешает реакции определени .However, the known method is laborious as there is only a small amount of phosphorylase in the serum with a large excess of other proteins, which interferes with the determination reaction.

Цель изобретени  - упрощение способа .The purpose of the invention is to simplify the method.

Поставленна  цель достигаетс  тем, что в сыворотке крови определ ют активность фосфорилазы В/ при этом сыворотку крови концентрируют путем мембранного фильтровани  или с помощьюОбработки нераствориг ым крахмалом, дл  чего пробу нанос т на колонку с крахмалом, при 4t предварительно набухшим в растворе, содержащем .фтористый натрий, / -глицерофосфат натри  и меркаптоэтанол при рН 6,1, после чего элюируют фосфорилазу раствором, содержащим (Ь -глицерофосфат натри , этилендиаминтетрауксусную кислоту и меркаптоэтанол , при РН 6,8 и t 30°, полученную фракцию инкубируют 60 мин при с раствором, содержащим гликоген, глюкозо-1-фосфат и аденозиниминфосфат в соотношении 5:3:2, полученную смесь обрабатыва эт трихлоруксусноП кислотой, образовавшиГ:с  осадок отдел ют, а в надосадочной жидкости определ ют фосфорилазу В The goal is achieved by determining the activity of phosphorylase B in the serum. The serum is concentrated by membrane filtration or by treatment with insoluble starch, for which a sample is applied to the column with starch, at 4t pre-swollen in solution containing. Fluoride sodium, sodium glycerophosphate and mercaptoethanol at pH 6.1, after which the phosphorylase is eluted with a solution containing (L sodium glycerophosphate, ethylenediaminetetraacetic acid and mercaptoethanol, at PH 6.8 and t 30 °, obtained The second fraction is incubated for 60 minutes with a solution containing glycogen, glucose-1-phosphate and adenosine-aminophosphate in a ratio of 5: 3: 2, the resulting mixture is treated with trichloroacetic acid to form the precipitate, and phosphorylase B is determined in the supernatant.

10 электрофоретическим методом и при наличии этого фермента диагносцируют инфаркт миокарда.10 electrophoretic method and in the presence of this enzyme diagnose myocardial infarction.

Пример 1 . Берут следующие средства дл  определени  фосфорилазы. Example 1 The following agents are used for the determination of phosphorylase.

15 Раствор Т. 0,1 М фторида натри  0,01 М натри -р-глицерофосфата ,рН 6,1 0,015 М меркаптоэтанола .15 Solution T. 0.1 M sodium fluoride 0.01 M sodium-p-glycerophosphate, pH 6.1 0.015 M mercaptoethanol.

2020

Раствор ГГ . 0,04 М натрн -Л-глицерофосфата , рН 6,8 0,001 М этилендиаминтетрауксусной кислоты 0,015 М меркаптоэта25 нола .GG solution. 0.04 M natrn-L-glycerophosphate, pH 6.8, 0.001 M ethylenediaminetetraacetic acid, 0.015 M mercapto ethe25 nola.

Татсвор mTatsvor m

0,04 М глицилглицина, рН 6,80.04 M glycylglycine, pH 6.8

0,01 М меркаптоэтанола 0,008 М этилендиамин- , 0.01 M mercaptoethanol 0.008 M ethylenediamine-,

30 тетрауксусной кислоты,30 tetraacetic acid,

Claims (1)

Формула изобретенияClaim Способ диагностики инфаркта миокарда путем определения активности фермента в сыворотке крови, отличающийся тем, что, с целью упрощения способа, в сыворотке крови определяют активность фосфорилазы В , при этом сыворотку крови концентрируют путем мембранного фильтрования или с помощью обработки нерастворимым крахмалом, для этого пробу наносят на колонку с крахмалом, при 4°C предварительно набухшим в растворе, содержащем фтористый натрий, β -глицерофосфат натрия и меркаптоэтанол при pH 6,1, после чего элюируют фосфорилазу раствором, содержащим β>-глицерофосфат натрия, этилендиаминтетрауксусную кислоту и меркаптоэтанол при pH 6,8 и t 30 С, полученную фракцию инкубируют 60 мин при 37°С с раствором, содержащим гликоген, глюкозо-1-фосфат и адено- . зиниминофосфат в соотношении 5:3:2, полученную смесь обрабатывают трихлоруксусной кислотой, образовавшийся осадок отделяют, а в надосадочной жидкости определяют фосфорилазу В электрофоретическим методом и при наличии этого фермента диагностируют инфаркт миокарда.A method for diagnosing myocardial infarction by determining the activity of an enzyme in blood serum, characterized in that, in order to simplify the method, the activity of phosphorylase B is determined in blood serum, while serum is concentrated by membrane filtration or by treatment with insoluble starch, for this the sample is applied to column with starch, at 4 ° C, previously swollen in a solution containing sodium fluoride, sodium β-glycerophosphate and mercaptoethanol at pH 6.1, after which the phosphorylase is eluted with a solution containing m β> sodium glycerophosphate, ethylenediaminetetraacetic acid, and mercaptoethanol at pH 6,8 and t 30 C, the resulting fraction was incubated 60 min at 37 ° C with a solution containing glycogen, glucose 1-phosphate and adenosine. ziniminophosphate in a ratio of 5: 3: 2, the resulting mixture is treated with trichloroacetic acid, the precipitate formed is separated, and phosphorylase B is determined in the supernatant by electrophoretic method and, in the presence of this enzyme, myocardial infarction is diagnosed.
SU741999385A 1973-02-07 1974-02-06 Method of diagnosis of myocardial infarction SU976958A1 (en)

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DD16876573A DD101986A1 (en) 1973-02-07 1973-02-07

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SU976958A1 true SU976958A1 (en) 1982-11-30

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BG (1) BG20730A1 (en)
CS (1) CS195756B1 (en)
DD (1) DD101986A1 (en)
DE (1) DE2401484A1 (en)
FR (1) FR2327545A1 (en)
GB (1) GB1451004A (en)
HU (1) HU169957B (en)
PL (1) PL94983B1 (en)
SU (1) SU976958A1 (en)

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BG20730A1 (en) 1975-12-20
FR2327545B1 (en) 1978-02-17
GB1451004A (en) 1976-09-29
FR2327545A1 (en) 1977-05-06
DD101986A1 (en) 1973-11-20
PL94983B1 (en) 1977-09-30
DE2401484A1 (en) 1974-08-29
CS195756B1 (en) 1980-02-29
HU169957B (en) 1977-03-28

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