Optimized Determination of γ-Glutamyltransferase by Reaction-Rate Analysis

1974 ◽  
Vol 20 (9) ◽  
pp. 1121-1124 ◽  
Author(s):  
Sidney B Rosalki ◽  
David Tarlow
Keyword(s):  
Dilute Hydrochloric Acid ◽  
Reaction Rate ◽  
Enzymatic Reaction ◽  
Enzyme Reaction ◽  
Enzyme Reactions ◽  
Rate Analysis ◽  
Acid Substrate ◽  
Incubation Buffer ◽  
Optimal Ph

Abstract We describe a method for measuring γ-glutamyltransferase (EC 2.3.2.2) activity in serum, which can be used with automated enzyme analyzers (such as the LKB 8600 Reaction Rate Analyzer) that require enzyme reactions to be initiated with substrate. The method also permits optimal determination conditions to be obtained at 37 °C. The enzymatic reaction is commenced by adding γ-glutamyl-p-nitroanilide dissolved in dilute hydrochloric acid to samples pre-incubated with tris(hydroxymethyl)aminomethane—glycylglycine buffer. The p-nitroaniline liberated is continously monitored at 37 °C at 405 nm. The pH of the pre-incubation buffer is such that the optimal pH for the enzyme reaction results from addition of the acid substrate solution.

Automated Enzymatic Determination of L-Lactate in Serum, with Use of a Miniature Centrifugal Anlyzer

1976 ◽  
Vol 22 (12) ◽  
pp. 2038-2041 ◽  
Author(s):  
T P Hadjiioannou ◽  
S I Hadjiioannou ◽  
S D Brunk ◽  
H V Malmstadt
Keyword(s):  
Lactate Dehydrogenase ◽  
Reaction Rate ◽  
Enzymatic Reaction ◽  
Reaction Rates ◽  
Enzymatic Determination ◽  
Analytical Recovery ◽  
Rate Method ◽  
Relative Errors ◽  
Lithium Lactate

Abstract We describe an automated enzymatic reaction-rate method for spectrophotometric determination of lactate in serum with a miniature centrifugal analyzer. The L(+)-lactate is selectively oxidized in the presence of lactate dehydrogenase (EC 1.1.1.27) and NAD+ to from NADH, whitch is measured from its absorption. Reaction rates are determined automatically, and unknown concentrations are calculated from a computer0generated calibration curve with aqueous lithium lactate standards. Lactte concentrations in the range 0.32-1.6 µg/4 µl (80-400 mg/liter) of sample were determined with relative errors and coefficient of variation of 4.8%. Analytical recovery of lactate added to pooled serum was 89-112% (average, 101%). Comparison with a kit ("Rapid Lactate") method gave a correlation coefficient squared of 0.979 over a concentration range of 39-779 mg/liter.

A stopped-flow clinical analyzer in which immobilized-enzyme reaction loops are used.

1977 ◽  
Vol 23 (6) ◽  
pp. 1033-1036 ◽  
Author(s):  
M D Joseph ◽  
D J Kasprzak ◽  
S R Crouch
Keyword(s):  
Immobilized Enzyme ◽  
Enzymatic Reaction ◽  
Enzyme Reaction ◽  
Fixed Time ◽  
Stopped Flow ◽  
Good Precision ◽  
Indicator Reaction ◽  
Flow Mixing ◽  
Kinetic Mode

Abstract A stopped-flow clinical analyzer is described that makes use of a reaction loop containing immobilized enzyme(s) for the determination of the analyte/substrate. The analyzer has been evaluated by determining glucose with immobilized glucose oxidase. The stopped-flow mixing system was constructed at a current cost of less than $500. The analyzer separates the enzymatic reaction from a followup, spectrophotometric indicator reaction. This separation allows the enzymatic reaction to be used in either a fixed-time, kinetic mode or in an equilibrium mode. Likewise, the indicator reaction can be used in either mode. Results for glucose in blood serum indicate that good precision and accuracy can be obtained.

Instabilities in a Simple Enzyme Reaction Caused by pH-Dependence

1995 ◽  
Vol 50 (12) ◽  
pp. 1147-1150 ◽  
Author(s):  
Gerold Baier ◽  
Peter Strasser ◽  
Ursula Kummer
Keyword(s):  
Reaction Rate ◽  
Ph Dependence ◽  
Enzymatic Reaction ◽  
Enzyme Reaction ◽  
Reaction Diffusion ◽  
Temporal Patterns ◽  
Homogeneous Reactor ◽  
Spatio Temporal

Abstract We investigate the model of an enzymatic reaction with Michaelis-Menten kinetics and bell-shaped pH-dependence of the reaction rate. In the case of proton consumption the reaction can generate oscillations in a homogeneous reactor and turbulent spatio-temporal patterns in a reaction-diffusion environment. I n s t a b il it i e s in a S im p le E n z y m e R e a c tio n C a u s e d b y p H -D e p e n d e n c e *

scholarly journals Determination of the enzyme reaction rate in a differential fixed-bed reactor: a case study

2001 ◽  
Vol 18 (1) ◽  
pp. 1-11 ◽  
Author(s):  
E.A. Baruque Filho ◽  
M.G.A. Baruque ◽  
G.L. Sant’Anna Jr.
Keyword(s):  
Reaction Rate ◽  
Fixed Bed ◽  
Enzyme Reaction ◽  
Fixed Bed Reactor

Automated enzymatic determination of L-lactate in serum, with use of a miniature centrifugal analyzer.

1976 ◽  
Vol 22 (12) ◽  
pp. 2042-2045 ◽  
Author(s):  
T P Hadjiioannou ◽  
S I Hadjiioannou ◽  
S D Brunk ◽  
H V Malmstadt
Keyword(s):  
Lactate Dehydrogenase ◽  
Reaction Rate ◽  
Enzymatic Reaction ◽  
Reaction Rates ◽  
Enzymatic Determination ◽  
Analytical Recovery ◽  
Rate Method ◽  
Relative Errors ◽  
Lithium Lactate

Abstract We describe an automated enzymatic reaction-rate method for spectrophotometric determination of lactate in serum with a miniature centrifugal analyzer. The L(+) -lactate is selectively oxidized in the presence of lactate dehydrogenase (EC 1.1.1.27) and NAD+ to form NADH, which is measured from its absorption. Reaction rates are determined automatically, and unknown concentrations are calculated from a computer-generated calibration curve with aqueous lithium lactate standards. Lactate concentrations in the range 0.32-1.6 mug/4 mul (80-400 mg/liter) of sample were determined with relative errors and coefficient of variation of 4.8%. Analytical recovery of lactate added to pooled serum was 89-112% (average, 101%). Comparison with a kit ("Rapid Lactate") method gave a correlation coefficient squared of 0.979 over a concentration range of 39-779 mg/liter.

Alcohol Determination by HPLC with Postcolumn Derivatization Based on the Thiosulfate-catalyzed Reaction of Alcohols and Cerium(IV) and Fluorescence Detection of Cerium(III)

2020 ◽  
Vol 16 ◽  
Author(s):  
Ikko Mikami ◽  
Eri Shibayama ◽  
Kengo Takagi
Keyword(s):  
Detection Limit ◽  
Fluorescence Detection ◽  
Reaction Rate ◽  
Detection Method ◽  
Fluorescence Detector ◽  
Reaction Solution ◽  
Optimum Detection ◽  
Postcolumn Derivatization ◽  
Sensitive Method

Background: Determination of a reducing substance based on the reaction between Ce(IV) and a reducing substance and fluorescence detection of Ce(III) generated has been reported as a selective and sensitive method. However, this method could not be applied to the determination of alcohol due to the low reaction rate of alcohol and Ce(IV). Objective: We found that thiosulfate catalytically enhanced reaction of alcohols (such as, methanol, ethanol, and propanol) and Ce(IV). Utilizing this effect, we developed a new method for the determination of alcohols. Results: In the presence of thiosulfate, an increase in fluorescence intensity was detected by injecting alcohol at concentrations of several millimolar, whereas it was not observed even at the concentration of 10% v/v (2 M for ethanol) in the absence of thiosulfate. The optimum detection conditions were determined to be 4.0 mM Ce(IV) sulfate and 0.50 mM thiosulfate, and the detection limit (S/N = 3) of ethanol under these conditions was 1 mM. In the calibration curves, changes in the slope were observed when the alcohol concentrations were approximately 10–25 mM. Using a thiosulfate solution containing ethanol as the reaction solution, a calibration curve without any change in slope was obtained, although the concentration of ethanol at the detection limit increased. The alcohols in the liquor and fuel were successfully analyzed using the proposed detection method as a postcolumn reaction. Conclusion: This new alcohol detection method using a versatile fluorescence detector can be applied to the postcolumn reaction of HPLC omitting need of time-consuming pretreatment processes.

Kinetics and mechanism of redox reactions of U(III) ions with monochloroacetic and dichloroacetic acids

1979 ◽  
Vol 44 (2) ◽  
pp. 401-405 ◽  
Author(s):  
Ľubica Adamčíková ◽  
Ľudovít Treindl
Keyword(s):  
Redox Reactions ◽  
Reaction Rate ◽  
Activation Parameters ◽  
Outer Sphere ◽  
Alcohol Concentration ◽  
Kinetics And Mechanism ◽  
Solvent Structure ◽  
Influence Of Ph ◽  
Sphere Mechanism

The kinetics and mechanism of the redox reactions of U3+ ions with mono- and dichloroacetic acids were studied. The influence of pH was observed mainly in the second case and led to the determination of the rate constants and activation parameters corresponding to two parallel steps, namely oxidation of U3+ with CHCl2COO- ions and oxidation of U3+ with CHCl2.COOH molecules. The influence of binary mixtures of water with methanol, ethanol, isopropanol, or tert-butenol on the reaction rate was followed. Increasing alcohol concentration influences the rate constant not only through changing dielectric constant and solvation of the reactants but also through a change of the solvent structure which plays a role in reactions with an outer sphere mechanism of the electron transfer.

Decomposition of peroxobenzoic acid in benzene catalyzed by transition metal 2,4-pentanedionates

1984 ◽  
Vol 49 (3) ◽  
pp. 673-679 ◽  
Author(s):  
Pavel Lederer ◽  
Eva Mácová ◽  
Josef Vepřek-Šiška
Keyword(s):  
Transition Metal ◽  
Reaction Rate ◽  
Reaction Products ◽  
Acid Decomposition ◽  
Analogous Experiment ◽  
Catalytic Effects

The decomposition of peroxobenzoic acid in benzene was studied, and catalytic effects of Fe(III), Mn(III), Co(II), Co(III), and Cr(III) on the reaction rate and the composition of the reaction mixture were investigated. An analogous experiment carried out in perdeuterobenzene and determination of the distribution of deuterium in the reaction products provided evidence for the participation of the solvent in peroxobenzoic acid decomposition.

Enzymatic colorimetry of lecithin and sphingomyelin in aqueous solution.

1983 ◽  
Vol 29 (8) ◽  
pp. 1513-1517 ◽  
Author(s):  
M W McGowan ◽  
J D Artiss ◽  
B Zak
Keyword(s):  
Aqueous Solution ◽  
Hydrogen Peroxide ◽  
Amniotic Fluid ◽  
Enzymatic Reaction ◽  
Detergent Solution ◽  
Enzymatic Determination ◽  
Red Color ◽  
Hydrolysis Of ◽  
Zwitterionic Detergent

Abstract A procedure for the enzymatic determination of lecithin and sphingomyelin in aqueous solution is described. The phospholipids are first dissolved in chloroform:methanol (2:1 by vol), the solvent is evaporated, and the residue is redissolved in an aqueous zwitterionic detergent solution. The enzymatic reaction sequences of both assays involve hydrolysis of the phospholipids to produce choline, which is then oxidized to betaine, thus generating hydrogen peroxide. The hydrogen peroxide is subsequently utilized in the enzymatic coupling of 4-aminoantipyrine and sodium 2-hydroxy-3,5-dichlorobenzenesulfonate, an intensely red color being formed. The presence of a non-reacting phospholipid enhances the hydrolysis of the reacting phospholipid. Thus we added lecithin to the sphingomyelin standards and sphingomyelin to the lecithin standards. This precise procedure may be applicable to determination of lecithin and sphingomyelin in amniotic fluid.

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