Automated measurement of amylase isoenzymes with 4-nitrophenyl-maltoheptaoside as substrate and use of a selective amylase inhibitor.
نویسندگان
چکیده
We automated a kinetic procedure for determining amylase isoenzymes in serum and urine samples. We used 4-nitro-phenylmaltoheptaoside as substrate and a selective amylase inhibitor with the Abbott-VP bichromatic system. By use of the maximum differences between pancreatic (P) and salivary (S) amylase activities remaining after inhibition by the selective inhibitor and by use of the linear range, a one-point standard method for calibration is proposed for determining amylase activities between about 50 and 1500 U/L when the P/S ratio exceeds 0.2. Results correlated well with those by electrophoresis and the Phadebas method (r = 0.99 for both pancreatic and salivary amylase). Reproducibilities (CVs) were 1.5% to 5.5% for pancreatic amylase and 1.4% to 3.3% for salivary amylase in serum, 0.8% to 2.0% for pancreatic amylase and 0.8% to 2.3% for salivary amylase in urine.
منابع مشابه
Automated measurement of amylase isoenzymes by a double kinetic assay with "blocked" beta-2-chloro-4-nitrophenyl maltopentaoside as substrate and with wheat germ inhibitor.
We evaluated the enzymic mechanism by which 3-keto butylidene-beta-2-chloro-4-nitrophenyl maltopentaoside (3KB-G5-CNP) serves as a substrate for serum pancreatic (p-) and salivary (s-) amylases. In aliquots of the reaction mixture, three kinds of beta-2-chloro-4-nitrophenyl oligosaccharides (glucose, maltoside, and maltotrioside) were separated from the substrate by high-performance liquid chro...
متن کاملMethods compared for determining total amylase activity and amylase isoenzymes in serum.
We evaluated two kinetic methods for determining total amylase activity and isoenzyme composition in serum. Stability studies of reagents for measuring total activity indicate that reagents containing 4-nitrophenyl-alpha-glucosides or enzyme-linked reagents can be stored only for seven days at 4 degrees C. Methods based on 4-nitrophenyl-alpha-glucoside substrates cannot be used if the reagent a...
متن کاملCatalytic concentrations of amylase isoenzymes: an assay with wheat-germ inhibitor and 4-nitrophenylmaltopentaoside plus 4-nitrophenylmaltohexaoside as substrate.
We coupled a kinetic procedure to a selective inhibiting method for determining amylase isoenzymes in biological samples, using 4-nitrophenylmaltopentaoside plus 4-nitrophenylmaltohexaoside as substrate and a wheat-germ selective inhibitor with the Gilford S-III spectrophotometer. On plotting remaining amylase activities/total amylase activities (R/T) vs pancreatic amylase activities/salivary a...
متن کاملReference Ranges for ot-Amylase in Serum and Urine with 4?6-Ethylidene-(G7)-l-4-nitrophenyl-(Gl)-a5D-maltoheptaoside äs Substrate
Reference ranges for -amylase in serum, spontaneously voided urine, and 24h urine were determined, using 4,6-ethylidene-(G7)-l-4-nitrophenyl-(Gl)-a,£)-malthoheptaoside äs the Substrate (EPS method), at 25, 30, and 37 °C. The measured values were evaluated with and without the use of a factor which converts the results of the -amylase EPS method into values comparable to those obtained with the ...
متن کاملEvaluation of silyl-blocked p-nitrophenylmaltoheptaoside as a substrate for alpha-amylase reagents.
We describe a reagent for measuring alpha-amylase (EC 3.2.1.1) activity in serum with use of a thexyldimethylsilyl ether of p-nitrophenyl-alpha-D-maltoheptaoside (SB7) as substrate. This substrate differs from Genzyme's benzylidene-blocked p-nitrophenylmaltoheptaoside substrate (B-PNPG7). The reagent, optimized for the characteristics of the silyl-blocked substrate, contains 4-(2-hydroxyethyl)-...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Clinical chemistry
دوره 30 7 شماره
صفحات -
تاریخ انتشار 1984