Determination of malonaldehyde in human plasma: elimination of spectral interferences in the 2-thiobarbituric acid reaction

The Analyst ◽  
1993 ◽  
Vol 118 (1) ◽  
pp. 89 ◽  
Author(s):  
Anunciaci�n Espinosa-Mansilla ◽  
Francisco Salinas ◽  
Amparo Rubio Leal
Keyword(s):  
Human Plasma ◽  
Thiobarbituric Acid ◽  
Spectral Interferences ◽  
Acid Reaction

Distribution of thiobarbituric acid-reactive substances in lipoproteins and proteins in serum.

1989 ◽  
Vol 35 (10) ◽  
pp. 2054-2058 ◽  
Author(s):  
D Bonnefont ◽  
A Legrand ◽  
J Peynet ◽  
J Emerit ◽  
J Delattre ◽  
...  
Keyword(s):  
Apolipoprotein B ◽  
Thiobarbituric Acid ◽  
Precipitation Method ◽  
High Density Lipoproteins ◽  
Separation Procedure ◽  
Serum Lipoproteins ◽  
Thiobarbituric Acid Reactive Substances ◽  
Selective Precipitation ◽  
Acid Reaction

Abstract We assessed the distribution of malondialdehyde (MDA) in lipoproteins and proteins in serum after using two procedures to separate the lipoproteins: sequential ultracentrifugation or selective precipitation with a sodium phosphotungstate and magnesium chloride reagent followed by ultracentrifugation of the supernate. MDA concentrations were determined by the thiobarbituric acid reaction and quantified by fluorometry. We found that 43% of the thiobarbituric acid-reactive substances (TBARS) was bound to the lipoproteins--27% to very-low- and low-density lipoproteins (VLDL-LDL) and 16% to high-density lipoproteins (HDL)--and from 11.5% to 15.8% to proteins, depending on the separation procedure. Residual unbound TBARS were located in the ultracentrifugation layers that contained no lipoproteins or proteins. The TBARS concentration in serum lipoproteins containing apolipoprotein B (i.e., VLDL-LDL) was the same after ultracentrifugation or selective precipitation. We therefore consider the precipitation method more suitable for routine TBARS determination in these lipoproteins, because it is easier to handle and faster. However, for determination of TBARS in HDL, selective precipitation requires subsequent ultracentrifugation at a density of 1.21 kg/L.

A rapid spectrofluorimetric method for the determination of malondialdehyde in human plasma after its derivatization with thiobarbituric acid and vortex assisted liquid–liquid microextraction

RSC Advances ◽  
2016 ◽  
Vol 6 (3) ◽  
pp. 2361-2367 ◽  
Author(s):  
Hossain Yahyavi ◽  
Massoud Kaykhaii ◽  
Mohammad Hashemi
Keyword(s):  
Human Plasma ◽  
Thiobarbituric Acid ◽  
Spectrofluorimetric Method ◽  
Liquid Microextraction

A rapid spectrofluorimetric method for the determination of malondialdehyde in human plasma after its derivatization with thiobarbituric acid and vortex assisted liquid–liquid microextraction.

A Simple Method for Estimation of Lipoxygenase and Cyclo-Oxygenase Pathways in Human Platelets - The Use of Thiobarbituric Acid Reaction

1979 ◽  
Author(s):  
M. Okuma ◽  
H. Takayama ◽  
H. Uchino
Keyword(s):  
Arachidonic Acid ◽  
Thin Layer ◽  
Normal Values ◽  
Lipid Peroxides ◽  
Thiobarbituric Acid ◽  
Myeloproliferative Disorders ◽  
Human Platelets ◽  
Simple Method ◽  
Acid Reaction

As lipid peroxides are produced from arachidonic acid(AA) by platelet lipoxygenase(PLO) and cyclo-oxygenase(PCO), a simple method was developed for estimation of these enzymic pathways by quantitating the thiobarbituric acid-reacting substance(TBARS) produced by the incubation of AA with human platelets. Thin-layer radiochromatographic analysis (J. Biol.Chem. 252:5871, 1977) of lipid products obtained by incubating [1-14C] AA with washed platelets under various conditions revealed that maximal amounts of PCO products were obtained by the incubation at 37°C within 1 min around pH 7.4, while those of PLO products within 10 min around pH 6.5. The incubation at 37°C of AA with aspirin-treated platelets at pH 7.4 for 1 min produced no TBARS, while that at pH 6.5 did TBARS as a function of the incubation time (up to 10 min). Based on these findings, PLO and PCO pathways were estimated by the determination of TBARS (J.Lab.Clin.Med. 75:283, 1970) obtained by incubating AA (0.2 mM, f.c.) either with aspirin-treated or with intact washed platelets (108) at 37°C as shown in the table. Normal values (M ± SD) expresse in terms of nmols of malonaldehyde were: PLO = 1. 76 ± 0 .36 (n=l7), PCO = 1.12 ± 0.42 (n=15). Alterations in these pathways were detecetd by this method in some patients with myeloproliferative disorders.

Studies on the Fibrinolytic System in Human Plasma: Quantitative Determination of Plasminogen Activators and Proactivators

1979 ◽  
Vol 41 (02) ◽  
pp. 365-383 ◽  
Author(s):  
C Kluft
Keyword(s):  
Pilot Study ◽  
Plasma Level ◽  
Human Plasma ◽  
Plasminogen Activators ◽  
Venous Occlusion ◽  
Quantitative Information ◽  
Optimal Recovery ◽  
Dextran Sulphate ◽  
Baseline Levels

SummaryEffects due to plasma plasminogen activators and proactivators are usually studied in assay systems where inhibitors influence the activity and where the degree of activation of proactivators is unknown. Quantitative information on activator and proactivator levels in plasma is therefore not availableStudies on the precipitating and activating properties of dextran sulphate in euglobulin fractionation presented in this paper resulted in the preparation of a fraction in which there was optimal recovery and optimal activation of a number of plasminogen activators and proactivators from human plasma. The quantitative assay of these activators on plasminogen-rich fibrin plates required the addition of flufenamate to eliminate inhibitors. The response on the fibrin plates (lysed zones) could be coverted to arbitrary blood activator units (BAU). Consequently, a new activator assay which enables one to quantitatively determine the plasma level of plasminogen activators and proactivators together is introduced.Two different contributions could be distinguished: an activity originating from extrinsic activator and one originating from intrinsic proactivators. The former could be assayed separately by means of its resistance to inhibition by Cl-inactivator. Considering the relative concentrations of extrinsic and intrinsic activators, an impression of the pattern of activator content in plasma was gained. In morning plasma with baseline levels of fibrinolysis, the amount of extrinsic activator was negligible as compared to the level of potentially active intrinsic activators. Consequently, the new assay nearly exclusively determines the level of intrinsic activators in morning plasma. A pilot study gave a fairly stable level of 100 ± 15 BAU/ml (n = 50). When fibrinolysis was stimulated by venous occlusion (15 min), the amount of extrinsic activator was greatly increased, reaching a total activator level of 249 ± 27 BAU/ml (n = 7).

A Simple Method for Estimation of Lipoxygenase and Cyclo-Oxygenase Pathways in Human Platelets - the Use of Thiobarbituric Acid Reaction

1980 ◽  
Vol 44 (02) ◽  
pp. 111-114 ◽  
Author(s):  
Hiroshi Takayama ◽  
Minoru Okuma ◽  
Haruto Uchino
Keyword(s):  
Time Course ◽  
Normal Values ◽  
Human Platelet ◽  
Thiobarbituric Acid ◽  
Human Platelets ◽  
Optimal Conditions ◽  
Experimental Conditions ◽  
Simple Method ◽  
Acid Reaction ◽  
Optimal Ph

SummaryTo develop a simple method for estimation of platelet lipoxygenase (PLO) and cyclo-oxygenase (PCO) pathways, the arachidonic acid (AA) metabolism of human platelet was investigated under various experimental conditions by the use of the thiobarbituric acid (TBA) reaction and a radioisotope technique. A TBA-reactive substance different from malondialdehyde (MDA) via PCO pathway was detected and shown to be derived from the PLO pathway. Since the optimal pH and time course of its formation were different from those of MDA formation via PCO pathway, PLO and PCO pathways were estimated by quantitating the TBA-reactive substances produced by the incubation of AA either with aspirin-treated platelets or with untreated ones, respectively, each under optimal conditions. Normal values expressed in terms of nmol MDA/108 platelets were 1.17±0.34 (M±SD, n = 31) and 0.79±0.15 (n = 31) for PLO and PCO pathways, respectively.

The Plasmin Inhibitors of Plasma

1964 ◽  
Vol 12 (01) ◽  
pp. 119-125 ◽  
Author(s):  
Y Shamash ◽  
A Rimon
Keyword(s):  
Human Plasma ◽  
New Method ◽  
Normal Amount ◽  
Caseinolytic Activity ◽  
Before And After ◽  
Plasmin Inhibitors

SummaryA new method for the assay of plasmin inhibitors in human plasma is described. The method consists of determination of the caseinolytic activity of a standard plasmin solution before and after incubation with the inhibitor, with lysine added to the mixture as a stabilizer of plasmin. Using this method, it was found that plasma contains enough inhibitors to inactivate 30 caseinolytic units of plasmin, or 10 times the normal amount of plasminogen in human plasma.

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