New, Simple Maltogenic Assay for Mechanized Determination of Alpha-Amylase Activity in Serum and Urine

1975 ◽  
Vol 21 (6) ◽  
pp. 694-702 ◽  
Author(s):  
Henning F Proelss ◽  
Billy W Wright
Keyword(s):  
Continuous Flow ◽  
Flow System ◽  
Amylase Activity ◽  
Alpha Amylase ◽  
Automated System ◽  
Normal Range ◽  
Clin Pathol ◽  
Reducing Substances ◽  
Serum And Urine

Abstract We report a new method for the mechanized determination of serum and urinary alpha-amylase by use of a continuous-flow system, based on the measurement of maltose formed by incubating the sample with amylodextrin at pH 7 and 40 °C. After dialysis, maltose is converted enzymatically to glucose, which is measured by Trinder's glucose oxidase—peroxidase method [J. Clin. Pathol. 22, 246 (1969)]. The reaction is linear for amylase activities up to 1400 Somogyi units/dl (2560 U/liter) and for maltose concentrations through 1500 mg/dl. No blank assay is required; consequently precision is improved and the automated system is simplified. Calibration with primary maltose standards increases accuracy and reliability. Common reducing substances in serum and urine do not interfere at their normal concentrations. There is a linear correlation between the results of this method and those of chromogenic and iodometric methods for normal and pathologic sera and urines. The chromogenic method yields significantly higher results and the iodometric method significantly lower results than this maltogenic method for elevated amylase activities. The normal range is 40-140 Somogyi units/dl (73-256 U/liter).

An Automated, Saccharogenic Method for Determining Serum Amylase Activity

1970 ◽  
Vol 16 (12) ◽  
pp. 985-989 ◽  
Author(s):  
Wendell R O'Neal ◽  
Nathan Gochman
Keyword(s):  
Glucose Oxidase ◽  
Blood Donors ◽  
Serum Amylase ◽  
Amylase Activity ◽  
Serum Glucose ◽  
Normal Range ◽  
Reducing Substances ◽  
Serum Amylase Activity

Abstract An automated adaptation of the Somogyi saccharogenic determination of serum amylase is described in which conventional AutoAnalyzer modules are used. Adequate sensitivity with short incubation is achieved by incorporating glucose oxidase and catalase in the substrate to destroy serum glucose during incubation. Maltose and other dialyzable oligosaccharides are measured with the alkaline copper-neocuproine reaction. A simultaneous blank run is performed to determine reducing substances other than glucose in serum. Precision studies and correlation with a manual saccharogenic method are presented. The normal range was determined from data for 49 healthy blood donors.

Continuous-flow potentiometric determination of α-amylase activity in serum and urine

1985 ◽  
Vol 32 (2) ◽  
pp. 183-190
Author(s):  
E.P. Diamandis ◽  
A. Papanastasiou-Diamandi ◽  
T.K. Christopoulos ◽  
T.P. Hadjiioannou
Keyword(s):  
Continuous Flow ◽  
Amylase Activity ◽  
Potentiometric Determination ◽  
Serum And Urine

New saccharogenic determination of alpha-amylase in serum and urine.

1979 ◽  
Vol 25 (2) ◽  
pp. 215-217 ◽  
Author(s):  
L van Leeuwen
Keyword(s):  
Human Serum ◽  
Amylase Activity ◽  
Glucose Dehydrogenase ◽  
Alpha Amylase ◽  
Endogenous Glucose ◽  
Alpha Glucosidase ◽  
Serum And Urine

Abstract I describe a new kinetic enzymatic saccharogenic method for assaying alpha-amylase in human serum and urine. alpha-Amylase liberates maltose from starch. This is successively acted on by alpha-glucosidase, mutarotase, and glucose dehydrogenase. The resulting conversion of NAD+ to NADH, measured at 340 nm, during a 20-min incubation reflects amylase activity. Endogenous glucose is destroyed before measurement of amylase activity is begun.

Study of the Saccharogenic Method for the Determination of Serum and Urine Amylase

1960 ◽  
Vol 6 (5) ◽  
pp. 434-452 ◽  
Author(s):  
Richard J Henry ◽  
Neil Chiamori
Keyword(s):  
Molecular Weight ◽  
Reaction Rate ◽  
Amylase Activity ◽  
Linear Reaction ◽  
Reducing Substances ◽  
Urine Amylase ◽  
Starch Substrate ◽  
Hydrolysis Of ◽  
Serum And Urine

Abstract A modification of Somogyi's saccharogenic method for the determination of amylase activity has been presented for serum and urine. The starch substrate employed is buffered sufficiently to control the pH of sera and urines. A study was made of the effect of varying enzyme and starch concentrations, pH, chloride and buffer concentrations, and temperature. Also studied were various starches and other substrates, several methods for quantitating reducing substances formed, stability of samples, precision, and normal values. Identification of intermediate and end products was made by paperchromatographic technics. Above a limiting concentration of starch, representing saturation of enzyme with substrate, a linear reaction rate was observed for a period of time that was found to be directly related to the initial starch concentration. Kjeldahl's law of proportionality has thus been confirmed for the ra-amylase activity of serum and urine. Evidence is presented that deviation from a linear reaction rate occurs at about the time the starch and high molecular weight dextrins no longer saturate the enzyme. Following this point, hydrolysis of lower molecular-weight sugars gradually becomes the preponderant reaction.

Enzyme-coupled ultraviolet determination of alpha-amylase activity with the GEMSAEC centrifugal analyzer.

1978 ◽  
Vol 24 (9) ◽  
pp. 1620-1624 ◽  
Author(s):  
W H Porter ◽  
R E Roberts
Keyword(s):  
Linear Response ◽  
Amylase Activity ◽  
Kinetic Studies ◽  
Alpha Amylase ◽  
Coefficients Of Variation ◽  
Endogenous Glucose ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Alpha Glucosidase ◽  
As Effects

Abstract We evaluated the Harleco alpha-glucosidase/hexokinase/glucose-6-phosphate dehydrogenase-coupled alpha-amylase method, bu use of the GEMSAEC centrifugal analyzer. Performance evaluation included kinetic studies of substrate and maltose hydrolysis as well as effects of endogenous glucose and fructose. The reagent was found to give a linear response with alpha-amylase activity to greater than 1200 U/liter. Within-run precision resulted in coefficients of variation (CV) of 0.9 to 3.2% over the range studied. Day-to-day precision corresponded to CV's of 2.4 to 4.4% over the same range of alpha-amylase procedure was found to be good (r = 0.997) for patients' sera examined.

Albumin activation of urinary amylase as determined with the Du Pont aca.

1978 ◽  
Vol 24 (4) ◽  
pp. 702-705 ◽  
Author(s):  
C C Garber ◽  
R N Carey
Keyword(s):  
Creatinine Clearance ◽  
Urine Analysis ◽  
Urinary Albumin ◽  
Alpha Amylase ◽  
Clearance Ratio ◽  
Activation Effect ◽  
Protein Activation ◽  
Urinary Amylase ◽  
Serum And Urine

Abstract Protein activation of urinary alpha-amylase (EC 3.2.1.1) activity was observed during an evaluation of the Du Pont aca procedure for the determination of urinary alpha-amylase. This activation effect became constant for urinary albumin concentrations exceeding 1.50 g/liter. It is recommended that urinary alpha-amylase be analyzed with sufficient albumin added to maximize this effect. The aca alpha-amylase procedure is compared to an amyloclastic method for both serum and urine analysis. Expected ranges are presented for the aca method for serum and urinary amylase, amylase clearance, and the amylase clearance/creatinine clearance ratio.

Automated Sample Cleanup for Pesticide Multiresidue Determination. I. Evaluation of Solvent Partitioning Module

1984 ◽  
Vol 67 (1) ◽  
pp. 108-111
Author(s):  
Fred M Gretch ◽  
Joseph D Rosen
Keyword(s):  
Cost Effectiveness ◽  
Column Chromatography ◽  
Continuous Flow ◽  
Flow System ◽  
Food Crops ◽  
System Data ◽  
The Cost ◽  
Automated Procedures ◽  
Multiresidue Determination

Abstract An automated continuous flow procedure is described that improves the cost effectiveness and precision of AOAC methodology for multiresidue pesticide determinations in nonfatty foods. Individual modules capable of performing automated solvent partitioning and automated column chromatography were constructed and integrated into a continuous flow system. Data are presented comparing the recoveries and precision for the determination of 8 pesticides (aldrin, dieldrin, p,p’ - DDT, ethion, heptachlor epoxide, lindane, parathion, and ronnel) partitioned from 2 food crops (spinach and tomatoes) by both the manual and automated procedures.

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