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Alpha Amylase CNPG3-Substrate


Alpha Amylase CNPG3-Substrate
The Alpha-amylases CNPG3-Substrate (1,4-a-D-glucanohydrolases, EC 3.2.1.1) catalyze the hydrolytic degradation of polymeric carbohydrates such as amylose, amylopectin and glycogen by cleaving 1,4-a-glucosidic bonds. In polysaccharides and oligosaccharides, several glycosidic bonds are hydrolyzed simultaneously. Maltotriose, the smallest such unit, is converted into maltose and glucose, albeit very slowly.

Two types of Alpha-amylases can be distinguished, the pancreatic type (P-type) and the salivary type (S-type). Whereas the P-type can be attributed almost exclusively to the pancreas and is therefore organ-specific, the S-type can originate from a number of sites. As well as appearing in the salivary glands it can also be found in tears, sweat, human milk, amniotic fluid, the lungs, testes and the epithelium of the fallopian tube.

Because of the sparsity of specific clinical symptoms of pancreatic diseases, a-amylase determinations are of considerable importance in pancreatic diagnostics. They are mainly used in the diagnosis and monitoring of acute pancreatitis. Hyperamylasemia does not, however, only occur with acute pancreatitis or in the inflammatory phase of chronic pancreatitis, but also in renal failure (reduced glomerular filtration), tumors of the lungs or ovaries, pulmonary inflammation, diseases of the salivary gland, diabetic ketoacidosis, cerebral trauma, surgical interventions or in the case of macroamylasemia. To confirm pancreatic specificity, it is recommended that an additional pancreas-specific enzyme - lipase or pancreatic-Alpha-amylase- also be determined.

Numerous methods have been described for the determination of Alpha-amylase. These either determine the decrease in the amount of substrate viscometrically, turbidimetrically, nephelometrically and amyloclastically or measure the formation of degradation products saccharogenically or kinetically with the aid of enzyme-catalyzed subsequent reactions.

The kinetic method described here is based on the cleavage of 2-chloro-4-nitrophenyl-αD-maltotrioside (CNP-G3) by a-amylase.

Intended use
Enzymatic in vitro test for the quantitative determination of a-amylase in human serum, plasma and urine.

Test principle
Colorimetric test with 2-chloro-4-nitrophenyl-αD-maltotrioside (CNP-G3) as direct substrate. Color is released directly as a result of a cleavage at the aglycone:

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