Determination of total and pancreatic alpha-amylase in human serum with 2-chloro-4-nitrophenyl-alpha-D-maltotrioside as substrate.
A reagent and assay conditions for the determination of the catalytic concentration of alpha-amylase (E.C. 3.2.1.1) in serum with 2-chloro-4-nitrophenyl-alpha-D-maltotrioside as substrate are described. The selected reaction mixture contains 50 mmol/l 2-(N-morpholino)ethanesulfonic acid buffer at pH 6.1 (37 degrees C), 300 mmol/l sodium chloride, 5 mmol/l calcium chloride and 450 mmol/l potassium thiocyanate. The described method is suitable for the measurement of total as well as pancreatic alpha-amylase by including antibodies against the salivary isoenzyme. The method shows the absence of a lag phase period, is sensitive and precise, has a large analytical range and is free from interference by hemoglobin, bilirubin and triglycerides. Comparative studies showed good correlation with other well established methods.
Gella
F J
FJ
Departament de Bioquimica i de Biologia Molecular, Universitat Autònoma de Barcelona, Bellaterra, Spain.
Gubern
G
G
Vidal
R
R
Canalias
F
F
D002021:Buffers; D002122:Calcium Chloride / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009596:Nitrophenols / Q000378:metabolism; D010179:Pancreas / Q000201:enzymology*; D012463:Saliva / Q000201:enzymology; D012680:Sensitivity and Specificity; D012965:Sodium Chloride / Q000378:metabolism; D013054:Spectrophotometry, Atomic; D013696:Temperature; D014312:Trisaccharides / Q000378:metabolism*; D000516:alpha-Amylases / Q000097:blood*
D002021:Buffers; D009596:Nitrophenols; D014312:Trisaccharides; C057413:2-chloro-4-nitrophenylmaltotrioside; C068311:4-chloro-2-nitrophenol; D012965:Sodium Chloride; D000516:alpha-Amylases; D002122:Calcium Chloride