Determination of pancreatic amylase activity by a radial diffusion assay

1970 ◽  
Vol 48 (11) ◽  
pp. 758-761 ◽  
Author(s):  
André Bélanger ◽  
Jean Perreault ◽  
Yvon Couture ◽  
Jacques Dunnigan
Keyword(s):  
Reaction Zone ◽  
Optical Density ◽  
Amylase Activity ◽  
Pancreatic Amylase ◽  
Insulin Hypoglycemia ◽  
Rat Pancreas ◽  
Iodine Complex ◽  
Radial Diffusion Assay ◽  
Diffusion Assay

A technique for assay of amylase content in homogenates of rat pancreas is described. It is based on the measurement of the diameter of a "reaction zone" resulting from the action of the enzyme diffusing through a gel containing the substrate. The method is used to evaluate the amylase content of the pancreas of rats stimulated by feeding and by insulin-hypoglycemia. The results are compared with those obtained by a classic amyloclastic method, using the optical density of the starch-iodine complex.

A Study of the Starch-Iodine Complex: A Modified Colorimetric Micro Determination of Amylase in Biologic Fluids

1964 ◽  
Vol 10 (10) ◽  
pp. 891-906 ◽  
Author(s):  
N R Pimstone
Keyword(s):  
Serum Amylase ◽  
Serum Proteins ◽  
Amylase Activity ◽  
Optimal Conditions ◽  
Serum Factor ◽  
Serum Factors ◽  
Iodine Complex ◽  
Human Sera ◽  
Initial Depression

Abstract The limitations, inaccuracies, and practical difficulties of saccharogenic methods are discussed. A modified colorimetric microdetermination of amylase is described in which the digestion of starch is measured by the decrease in the starch-iodine color. Experimental data show that there are two other serum factors that can also cause a fall-off in color: (1) an immediate 10-15% depression of color, probably due to serum proteins and countered by using serum in the control; (2) an acid-serum factor causing a progressive fall-off in color subsequent to the initial depression. Iodine prevents this, and must be added as soon as the acid has been added to stop the enzyme activity. Results of 189 consecutive assays of human sera are presented. Amylase activity of duodenal aspirate has been determined simultaneously by the method described and the Lagerlöf method. Results are compared. Changes in serum amylase and lipase levels in artificially produced pancreatitis in dogs are presented. Optimal conditions for amylase activity are reviewed, and in the light of these, different amyloclastic methods and their results compared. Achroic-point technics are briefly evaluated.

SOME OBSERVATIONS ON THE PANCREATIC AMYLASE AND INTESTINAL MALTASE OF THE CHICK

1963 ◽  
Vol 41 (1) ◽  
pp. 2107-2121 ◽  
Author(s):  
B. M. Laws ◽  
J. H. Moore
Keyword(s):  
Small Intestine ◽  
Amylase Activity ◽  
Pancreatic Amylase ◽  
Ph Optimum ◽  
Modified Method ◽  
Mucosal Cells ◽  
Small Intestinal ◽  
The Stability ◽  
Hydrolysis Of

The digestive enzymes amylase and maltase were studied in acetone-dried powders or homogenates of the pancreatic and small intestinal tissues and small intestinal contents obtained from chicks of various ages. The stability of pancreatic amylase, which was relatively low in 0.15 M sodium chloride, was increased markedly by the presence of 0.02 M barbiturate buffer. The pH optimum of pancreatic amylase (chloride-activated) was 7.0 whereas that of intestinal maltase was 6.9. High levels of pancreatic amylase activity were found in the newly-hatched chick but these levels decreased during the following 20 days and then remained constant. The contrast between the high amylase and low maltase activities in the contents of the small intestine suggested that molecules of maltose, formed by the hydrolysis of starch, were absorbed as such by the mucosal cells. It appeared that maltose could be absorbed with equal facility from all sections of the small intestine of the 10-day-old chick but in the older birds maltose absorption seemed to occur more readily from the upper small intestine than from the duodenum and lower small intestine. A modified method for the determination of maltase activity is described.

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.

Determination of amylase activity in serum by using a wheat germ inhibitor with the Du Pont aca.

1986 ◽  
Vol 32 (8) ◽  
pp. 1539-1541 ◽  
Author(s):  
D A Lacher ◽  
M B Harize
Keyword(s):  
Acute Pancreatitis ◽  
Wheat Germ ◽  
Serum Amylase ◽  
Amylase Activity ◽  
Reference Interval ◽  
Pancreatic Amylase ◽  
Wheat Germ Extract ◽  
Rapid Procedure ◽  
Assay Conditions

Abstract A rapid procedure for determining salivary- and pancreatic-type amylase (EC 3.2.1.1) in serum by incorporating a wheat germ inhibitor (from Triticum aestivum) was developed for the Du Pont aca IV analyzer. Under optimal assay conditions, activities of salivary and pancreatic amylase were inhibited by 93% and 19%, respectively. The 95% central reference interval for the percentage of inhibition of serum amylase was 38-84%. Patients with acute pancreatitis showed less than 26% inhibition of amylase after addition of the wheat germ extract, reflecting the prevalence of pancreatic-type amylase in this disorder.

scholarly journals Total and Pancreatic Amylase Measured with 2-Chloro-4-nitrophenyl-4-O-β-d-galactopyranosylmaltoside

2000 ◽  
Vol 46 (7) ◽  
pp. 928-933 ◽  
Author(s):  
Yoshitaka Morishita ◽  
Yoshitsugu Iinuma ◽  
Nobuo Nakashima ◽  
Keiichi Majima ◽  
Katsuhiko Mizuguchi ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Amylase Activity ◽  
Biological Fluids ◽  
Transfer Reactions ◽  
Pancreatic Amylase ◽  
Salivary Amylase ◽  
Specific Determination ◽  
Routine Procedures

Abstract Background: Many different methods have been used to assay amylase activity, using nitrophenylated oligosaccharides as substrate; however, the hydrolysis steps in these methods are complex. Methods: We developed a new continuously monitoring assay for amylase activity in biological fluids, using 2-chloro-4-nitrophenyl-4-O-β-d-galactopyranosylmaltoside (GalG2CNP) as the substrate; this assay was used with anti-human salivary amylase monoclonal antibodies for specific determination of the pancreatic isoenzyme. Amylase converted GalG2CNP into β-d-galactopyranosylmaltose and 2-chloro-4-nitrophenol, which was measured at 405 nm. Results: GalG2CNP was cleaved between 2-chloro-4-nitrophenol and β-d-galactopyranosylmaltose and did not undergo transfer reactions. The within-assay CVs (n = 20) for total amylase (T-AMY) and pancreatic amylase (P-AMY) were 0.6–1.6% and 0.5–2.5%, respectively; and day-to-day CVs (n = 10) for T-AMY and P-AMY were 0.8–3.7% and 0.6–4.1%, respectively. T-AMY and P-AMY activities in serum or urine obtained by the proposed method correlated well with those determined by the 2-chloro-4-nitrophenyl 4-O-β-d-galactopyranosyl-β-maltotetraoside method or the modified IFCC method. Conclusions: This novel assay for T-AMY and P-AMY measures both activities stoichiometrically, directly, and easily, and may be suitable for routine procedures.

SOME OBSERVATIONS ON THE PANCREATIC AMYLASE AND INTESTINAL MALTASE OF THE CHICK

1963 ◽  
Vol 41 (10) ◽  
pp. 2107-2121 ◽  
Author(s):  
B. M. Laws ◽  
J. H. Moore
Keyword(s):  
Small Intestine ◽  
Amylase Activity ◽  
Pancreatic Amylase ◽  
Ph Optimum ◽  
Modified Method ◽  
Mucosal Cells ◽  
Small Intestinal ◽  
The Stability ◽  
Hydrolysis Of

The digestive enzymes amylase and maltase were studied in acetone-dried powders or homogenates of the pancreatic and small intestinal tissues and small intestinal contents obtained from chicks of various ages. The stability of pancreatic amylase, which was relatively low in 0.15 M sodium chloride, was increased markedly by the presence of 0.02 M barbiturate buffer. The pH optimum of pancreatic amylase (chloride-activated) was 7.0 whereas that of intestinal maltase was 6.9. High levels of pancreatic amylase activity were found in the newly-hatched chick but these levels decreased during the following 20 days and then remained constant. The contrast between the high amylase and low maltase activities in the contents of the small intestine suggested that molecules of maltose, formed by the hydrolysis of starch, were absorbed as such by the mucosal cells. It appeared that maltose could be absorbed with equal facility from all sections of the small intestine of the 10-day-old chick but in the older birds maltose absorption seemed to occur more readily from the upper small intestine than from the duodenum and lower small intestine. A modified method for the determination of maltase activity is described.

PERMISSIVE ROLE OF l-THYROXINE IN INDUCTION OF PANCREATIC AMYLASE BY CORTISOL IN NEONATAL RATS

1980 ◽  
Vol 86 (3) ◽  
pp. 497-500 ◽  
Author(s):  
MASAYOSHI KUMEGAWA ◽  
NORIHIKO MAEDA ◽  
TOSHIHIKO YAJIMA ◽  
TAISHIN TAKUMA ◽  
EIKO IKEDA ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Amylase Activity ◽  
Daily Injection ◽  
Pancreatic Amylase ◽  
Neonatal Rats ◽  
Rat Pancreas ◽  
Developing Rat ◽  
Permissive Role ◽  
Intact Rats

The effect of l-thyroxine (T4) on amylase activity in the developing rat pancreas has been investigated. Administration of T4 (0·2 μg/g body wt) alone to intact rats on days 5–10 after birth did not induce pancreatic amylase but the enzyme was induced significantly by daily injection of cortisol (10 μg/g body wt) alone into intact rats over the same period. In thyroidectomized, adrenalectomized rats pancreatic amylase was not induced by the injection of cortisol alone but it was induced by the administration of cortisol plus T4. Increase in enzyme activity was much less in thyroidectomized animals than in intact animals. These results suggested that T4 does not have a direct effect in increasing pancreatic amylase activity but plays a permissive role in increasing enzyme activity.

Three methods compared for determination of pancreatic and salivary amylase activity in serum of cystic fibrosis patients.

1986 ◽  
Vol 32 (2) ◽  
pp. 296-300 ◽  
Author(s):  
R O Wolf ◽  
V S Hubbard ◽  
B K Gillard ◽  
A Kingman
Keyword(s):  
Cystic Fibrosis ◽  
Gel Electrophoresis ◽  
Serum Amylase ◽  
Amylase Activity ◽  
Pancreatic Insufficiency ◽  
Polyacrylamide Gel Electrophoresis ◽  
Pancreatic Amylase ◽  
Salivary Amylase ◽  
Amylase Isoenzymes

Abstract We evaluated three methods for serum amylase (EC 3.2.1.1) isoenzymes to determine whether they are interchangeable and to test their ability to discriminate between cystic fibrosis patients with and without pancreatic insufficiency. One method involved salivary amylase inhibitor (O), and two were polyacrylamide gel electrophoresis separations differing in method of detection--either direct zymogram (G) or gel slicing followed by activity estimates per slice (W). Results for percentage pancreatic amylase differed significantly. Reproducibility for percentage pancreatic amylase was high, moderate, and low (r = 0.95, 0.53, and 0.02) for methods G, O, and W, respectively; moderate (r = 0.60) among the three methods; and moderate between pairs. Therefore, this result for a subject must be considered relative to the method used in its determination. The clinical diagnosis of pancreatic insufficiency was verified by 77.8%, 83.3%, and 94.4% correct classification rates for methods O, W, and G, respectively. Evidently, method G is the most efficient and may be the method of choice for measuring serum amylase isoenzymes in cystic fibrosis.

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