SU1091071A1 - Method of determination of monoaminoxydase activity in trombocytes - Google Patents

Method of determination of monoaminoxydase activity in trombocytes Download PDF

Info

Publication number
SU1091071A1
SU1091071A1 SU823491323A SU3491323A SU1091071A1 SU 1091071 A1 SU1091071 A1 SU 1091071A1 SU 823491323 A SU823491323 A SU 823491323A SU 3491323 A SU3491323 A SU 3491323A SU 1091071 A1 SU1091071 A1 SU 1091071A1
Authority
SU
USSR - Soviet Union
Prior art keywords
method
activity
determination
ml
monoaminoxydase
Prior art date
Application number
SU823491323A
Other languages
Russian (ru)
Inventor
Александр Зосимович Дроздов
Борис Михайлович Коган
Original Assignee
Всесоюзный Ордена Трудового Красного Знамени Научно-Исследовательский Институт Общей И Судебной Психиатрии Им.В.П.Сербского
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Всесоюзный Ордена Трудового Красного Знамени Научно-Исследовательский Институт Общей И Судебной Психиатрии Им.В.П.Сербского filed Critical Всесоюзный Ордена Трудового Красного Знамени Научно-Исследовательский Институт Общей И Судебной Психиатрии Им.В.П.Сербского
Priority to SU823491323A priority Critical patent/SU1091071A1/en
Application granted granted Critical
Publication of SU1091071A1 publication Critical patent/SU1091071A1/en

Links

Abstract

METHOD FOR DETERMINING THE ACTIVITY OF MONOAMINOXIDASE IN THROMBOCYTES, including their incubation in the presence of benzylamine followed by spectrophotometric determination of the concentration of benzaldehyde formed, characterized by TeMj, which, in order to be precise, isoaldehyde extracted with N-methanol

Description

o The invention relates to medicine in particular to the enzyme analysis of human blood, and can be used in medical practice in the study of blood biochemical parameters in the clinic and in experiment. A known method for the fluorometric determination of monoamine oxidase (MAO) activity in platelets is carried out by incubating the enzyme in question with a substrate. The determination is carried out in 0.2 MJ of blood after incubating the reaction mixture for 1 hour at fij. The disadvantages of this method are the low linearity range and low specificity. In addition, the use of 1,2-diaminonaphthalene substances with strong carcinogenic substances as reagents, gives the method greater trauma. A method of spectrophotometric measurement of enzyme activity is also known, including the extraction of bexaldehyde with cyclohexane followed by detection with 242 to them 2. A disadvantage of the known method is the relatively low accuracy. The purpose of the invention is to improve the accuracy of the method. The goal is achieved by the method of determining the activity of α-monoamine oxidase in platelets, including incubating them in the presence of benzylamine followed by spectrophotometric determination of the concentration of benzaldehyde formed, the benzaldehyde is extracted with n-heptane in a ratio of (1: 2.1) - (1: 2, 6), the chromatog extract is refined at the flow rate of eluent 1.9-2.1 ml / min, and a mixture of ethanol and WATER is used as eluent in the ratio (58:42) - (62:38 The method is carried out as follows. are from 10 ml kr Ova taken from the cubital vein into plastic cups, to which 0.3 ml of 0.27 M EDTA buffer, pH 7.4, are previously added. Red blood cells and white blood cells are precipitated by centrifuging the blood for 5 minutes at 300 g. centrifuged for 20 minutes at 2000 g. The precipitate is suspended and homogenized in 0.01 M phosphate buffer pH 7.65, the homogenate is centrifuged at 20,000 g for 20 minutes the precipitate is suspended and homogenized in 0.2 M phosphate buffer pH 8.4. 0.1 ml of the suspension is taken into the reaction medium. The amount of protein is determined by the method of Lowry and .0.4 ml of 0.2 M K-Na phosphate buffer pH 8.4 is added. Then, 0.1 ml (12 mmol) of the benzylamine solution is added to the k-reaction mixture, and the samples are incubated at 37 ° C for 10 minutes with constant shaking. The reaction is stopped with 0.1 ml of 20% TCA. Following this, heptane is added to the reaction mixture in a ratio of 1: 2.1-1-1: 2.6. The tubes are shaken vigorously for 3 minutes and centrifuged at 1500 g for 15 minutes. The organic phase is transferred to the autosampler vials, from where it enters the chromatograph loop (AnaIyst-7800 high pressure liquid chromatograph, manufactured by LDC). The volume of solution injected into the column with reversed phase (, 6 mm Sperisorb ODS-10) is 20 µl. As the mobile phase using a mixture of ethanol with water in the ratio of 58: 42-62: 38. The flow rate of eluent is 1.9-2.1 ml / min. Example 1. Test M.. The collection of blood samples, the isolation of thrombogocytes, and their incubation with benzylamine is carried out in the manner described. The resulting benzaldehyde is extracted with H-heptane (1: 2.1), then chromatographed at a flow rate of 1.9 ml / min. An ethanol-water mixture (58:42) is used as the mobile phase. The specific enzyme activity was 73.33 mmol / h / mg protein. Example 2 Subject B. Sampling of blood, selection of platelets, incubation with benzylamine is carried out as described. The resulting benzaldehyde is extracted with H-heptane (1: 2.6), then chromatographed at a flow rate of 2.1 ml / min. A mixture of ethanol water (62:38) was used as the mobile phase, while the specific activity of the enzyme was 65.28 mmol / h / mg protein.

31091071

Using the proposed method of analysis up to 94%. In addition, the method of ba for determining the activity of MAO in thromscine is easy to use; it does not require bocytes, which will increase the accuracy of inaccessible chemicals.

Claims (1)

  1. METHOD FOR DETERMINING THE ACTIVES OF MONOAMINOXIDASE IN THROMBOCYTHES : 2.6), the extract is chromatographed at the flow rate of eluent 1.9-2.1, ml / min, and a mixture of ethanol and water is used as an eluent in the ratio (58:42) - (62:38).
    8i, "1091071
    1091071
SU823491323A 1982-09-13 1982-09-13 Method of determination of monoaminoxydase activity in trombocytes SU1091071A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
SU823491323A SU1091071A1 (en) 1982-09-13 1982-09-13 Method of determination of monoaminoxydase activity in trombocytes

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
SU823491323A SU1091071A1 (en) 1982-09-13 1982-09-13 Method of determination of monoaminoxydase activity in trombocytes

Publications (1)

Publication Number Publication Date
SU1091071A1 true SU1091071A1 (en) 1984-05-07

Family

ID=21029104

Family Applications (1)

Application Number Title Priority Date Filing Date
SU823491323A SU1091071A1 (en) 1982-09-13 1982-09-13 Method of determination of monoaminoxydase activity in trombocytes

Country Status (1)

Country Link
SU (1) SU1091071A1 (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5906570A (en) * 1995-04-18 1999-05-25 Cobe Laboratories, Inc. Particle filter apparatus
US7708889B2 (en) 2002-04-16 2010-05-04 Caridianbct, Inc. Blood component processing system method

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
1. Мс. Ewen.jCh. М. Colen G.B. An amlne oxidase in normal human.J. Lab. Chim. Med., 62(Б), 766776, 1963. 2. Zaitsu K., Naga H., Ohtsubo K.et.al. A Sensitive Fluorometric Assay of Human, Platelet Monoamine Oxidase inhibitor. - Chem. Plarm. Bull., 1978, 26, p. 34713476 (прототип). *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5906570A (en) * 1995-04-18 1999-05-25 Cobe Laboratories, Inc. Particle filter apparatus
US5913768A (en) * 1995-04-18 1999-06-22 Cobe Laboratories, Inc. Particle filter apparatus
US5951877A (en) * 1995-04-18 1999-09-14 Cobe Laboratories, Inc. Particle filter method
US7708889B2 (en) 2002-04-16 2010-05-04 Caridianbct, Inc. Blood component processing system method

Similar Documents

Publication Publication Date Title
Schirch et al. Serine transhydroxymethylase identification as the threonine and allothreonine aldolases
Evans Manual and automated methods for measuring urea based on a modification of its reaction with diacetyl monoxime and thiosemicarbazide.
US3466249A (en) Blood serum reference standard for multi-automated analytical procedures
Lever et al. The estimation of renin in plasma by an enzyme kinetic technique
Neeld Jr et al. Macro-and micromethods for the determination of serum vitamin A using trifluoroacetic acid
Marciniak et al. Autoprothrombin C: A second enzyme from prothrombin
Brown et al. A colorimetric micromethod for determination of ammonia; the ammonia content of rat tissues and human plasma
Dennog et al. Detection of DNA damage after hyperbaric oxygen (HBO) therapy
Loo et al. Assay of streptomycin by the paper-disc plate method
Herbert et al. Crystalline bacterial catalase
Heatley A method for the assay of penicillin
Norman Studies of the plasmin system: I. Measurement of human and animal plasminogen. Measurement of an activator in human serum
Palmer et al. Heterogeneity of rabbit antibody and its subunits
Chiknas A liquid chromatography-assisted assay for angiotensin-converting enzyme (peptidyl dipeptidase) in serum.
Foster et al. Stable reagents for determination of serum triglycerides by a colorimetric Hantzsch condensation method
Qureshi et al. Application of high-performance liquid chromatography to the determination of free amino acids in physiological fluids
Switzer et al. Improved method for hydroxyproline analysis in tissue hydrolyzates
Lund et al. Association-dissociation behavior and subunit structure of heat-released nitrate reductase from Escherichia coli.
Boyce et al. Ion-binding properties of electrophoretically homogeneous mucoproteins of urine in normal subjects and in patients with renal calculus disease
Karl et al. A microfluorometric enzymatic assay for the determination of alanine and pyruvate in plasma and tissues
FR2325920B1 (en)
TW592662B (en) Biological fluid constituent sampling and measurement devices and methods
Forrester et al. Enzymatic method for determination of CO2 in serum.
Phillips Rapid analysis of inflammatory cytokines in cerebrospinal fluid using chip‐based immunoaffinity electrophoresis
Cohen et al. The polyamine content of the tRNA of E. coli