Clinical Chemistry: Determination of Amylase Activity and Amylase Isoenzymes in Serum and Urine Using a Solid Phase Blue Starch Substrate

1975 ◽  
Vol 35 (2) ◽  
pp. 163-169 ◽  
Author(s):  
Kaarina Ojala ◽  
A. Harmoinen
Keyword(s):  
Solid Phase ◽  
Amylase Activity ◽  
Clinical Chemistry ◽  
Amylase Isoenzymes ◽  
Starch Substrate ◽  
Serum And Urine

Determination of amylase activity in serum and urine using blue starch substrate

1972 ◽  
Vol 42 (1) ◽  
pp. 63-66 ◽  
Author(s):  
Irie Akiko ◽  
Hunaki Masaaki ◽  
Bando Keiichi ◽  
Kawai Kazuo
Keyword(s):  
Amylase Activity ◽  
Starch Substrate ◽  
Serum And Urine

Electrophoretic pattern of amylase isoenzymes in serum and urine of normal persons.

1976 ◽  
Vol 22 (4) ◽  
pp. 439-444 ◽  
Author(s):  
M Otsuki ◽  
S Saeki ◽  
H Yuu ◽  
M Maeda ◽  
S Baba
Keyword(s):  
Thin Layer ◽  
Human Serum ◽  
Salivary Glands ◽  
Clinical Medicine ◽  
Amylase Activity ◽  
Electrophoretic Pattern ◽  
Normal Pattern ◽  
Amylase Isoenzymes ◽  
Serum And Urine

Abstract We separated and measured amylase isoenzymes in the serum and urine of 3036 normal persons by electrophoresis on a thin layer of polyacrylamide gel. We wished to establish the normal pattern of these isoenzymes and to evaluate the usefulness of this method of electrophoresis in clinical diagnosis. Results for patients with hyper- or hypofunctioning pancreas and salivary glands suggested that essentially all the isoamylases in human serum and urine are derived from the salivary glands and the pancreas, and revealed that isoamylases of more than 98% of normal persons consisted of two major isoenzymes and two to three minor ones. Although these observations indicate that data on changes in the proportion of amylase activity of each isoenzyme can be useful in clinical medicine, the following points should be remembered: (a) quantitative differences in the isoenzyme pattern were observed, depending upon the condition of the samples; (b) because the proportion of isoamylase activity in serum of different normal persons differs, seriatim determination of amylase isoenzymes is necessary; and (c) because five different genetically controlled types of isoamylases were observed in normal persons, genetic investigations are also necessary.

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.

Isopropylidine maltoheptosyl fructofuranoside, doubly blocked substrate for determination of endoamylase activity

1991 ◽  
Vol 37 (8) ◽  
pp. 1323-1328
Author(s):  
Z Ogawa ◽  
Y Matsubayashi ◽  
S Satoh ◽  
N Orita ◽  
H Itoh
Keyword(s):  
Human Serum ◽  
Coefficient Of Variation ◽  
Oxidation Rate ◽  
Amylase Activity ◽  
Mannitol Dehydrogenase ◽  
Coupled Reactions ◽  
Alpha Glucosidase ◽  
Serum And Urine

Abstract We synthesized o-(4,6-o-isopropylidene-alpha-D-glucopyranosyl)-(1----4)- [o-alpha-D-glucopyranosyl-(1----4])5-o-alpha-D-glucopyranosyl-(1----2)- alpha-D-fructofuranoside (IPG7F) and developed an assay for determining the activity of amylase in human serum and urine by using this substrate. Glucoamylase, alpha-glucosidase, and mannitol dehydrogenase are used as coupling enzymes. The coupled reactions are monitored by continuously measuring the oxidation rate of NADH. In this procedure, various substances in the test specimens do not interfere with the detection of amylase activity. Exactly one molecule of NADH is oxidized by one attack of amylase on the substrate, although four products can be produced in the reaction. The within-assay coefficient of variation (CV) ranged from 1.0% to 4.1% and the between-assay CV ranged from 2.6% to 5.3%. The results of our new assay correlate well with those of the amylase assay involving p-nitrophenol maltoheptaoside as substrate (r = 0.978) and with those of the amylase assay involving maltopentaose (r = 0.987).

Specific immunoassay of alpha-amylase isoenzymes in human serum.

1985 ◽  
Vol 31 (8) ◽  
pp. 1331-1334 ◽  
Author(s):  
M Gerber ◽  
K Naujoks ◽  
H Lenz ◽  
W Gerhardt ◽  
K Wulff
Keyword(s):  
Monoclonal Antibody ◽  
Human Serum ◽  
Enzyme Immunoassay ◽  
Amylase Activity ◽  
Alpha Amylase ◽  
Pancreatic Amylase ◽  
Routine Method ◽  
Salivary Amylase ◽  
Amylase Isoenzymes

Abstract A monoclonal antibody (66C7) was prepared that specifically binds human salivary amylase (EC 3.2.1.1); it cross reacts with human pancreatic amylase by less than 1%. Two procedures are described for determination of isoamylases in human serum with this antibody: an enzyme immunoassay for determining amylase of salivary origin, and a routine method in which this amylase is immunoprecipitated and the remaining (pancreatic) amylase activity is assayed. Results by the two methods correlate well.

Maltotetraose as a substrate for enzyme-coupled assay of amylase activity in serum and urine.

1979 ◽  
Vol 25 (3) ◽  
pp. 481-483 ◽  
Author(s):  
K J Whitlow ◽  
N Gochman ◽  
R L Forrester ◽  
L J Wataji
Keyword(s):  
Amylase Activity ◽  
Biological Fluids ◽  
Urine Samples ◽  
Coupled Assay ◽  
Serum And Urine ◽  
Serum And Urine Samples

Abstract The use of maltotetraose ss a new substrate for the enzyme-coupled determination of amylase activity in biological fluids was developed by Beckman Microbics. We evaluated a manual and a centrifugal analyzer version of the method in comparison with two commonly used manual starch-dye amylase techniques: Roche Amylochrome and Pharmacia Phadebas. Both maltotetraose amylase procedures proved to be rapid and precise, and results correlated satisfactorily with the starch-dye methods for serum and urine samples.

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