Biosystems Pancreatic Alpha Amylase
PRINCIPLE OF THE METHOD
Amylase catalyzes the hydrolysis of 4-nitrophenyl-maltoheptaoside-ethylidene to smaller oligosacharides which are hydrolyzed by -glucosidase liberating 4-nitrophenol.
The catalytic concentration is determined from the rate of 4-nitrophenol formation, measured at 405 nm1,2
Specific antibodies inhibits the salivary isoenzyme and thus allow the measurement of pancreatic α-amylase3,4
CONTENTS AND COMPOSITION
A. Reagent: 1 x 20 mL. HEPES 50 mmol/L, calcium chloride 0.075 mmol/L, sodium chloride 90 mmol/L, magnesium chloride 13 mmol/L, -glucosidase > 4 U/mL, pH 7.1, monoclonal
antibodies (mouse) 50 mg/L.
B. Reagent: 1 x 5 mL. HEPES 50 mmol/L, 4-Nitrophenyl-maltoheptaoside-ethylidene 18 mmol/L, pH 7.1.
Store at 2-8ºC.
Reagents are stable until the expiry date shown on the label when stored tightly closed and if
contaminations are prevented during their use.
Indications of deterioration:
Reagents: Presence of particulate material, turbidity, absorbance of the blank over 0.300 at
405 nm (1 cm cuvette).
Reagents are provided ready to use.
Analyzer, spectrophotometer or photometer with cell holder thermostatable at 37ºC and able to read at 405 nm.
Cuvettes with 1 cm light path.
Serum, plasma or urine collected by standard procedures.
Pancreatic -amylase in serum or plasma is stable for 1 month at 2-8ºC. Use heparin or EDTA
Pancreatic -amylase in urine is stable for 1 month at 2-8ºC if pH is adjusted to approximately 7
before storage. Centrifuge or filter before testing.
1. Bring the Reagent and the instrument to reaction temperature.
2. Pipette into a cuvette: Notes 1,2
CONTENTS AND STRUCTURE
- Reagent: 1 x 20 ml. HEPES of 50 mmol/l, calcium chloride of 0.075 mmol/l, sodium chloride of 90 mmol/l, magnesium chloride of 13 mmol/l, a-glyukozidaza> 4 Eml, antibodies monoclonal (mouse) of 50 mg/l, pH 7.1.
- Reagent: 1 x 5 ml. HEPES of 50 mmol/l, 4-nitrophenyl-maltogeptaozid-etiliden of 18 mmol/l, pH 7.1.
– The analyzer, the spectrophotometer or a photometer with thermostatically controlled kyuvety at 37ºC for removal of indications at 405 nanometers.
– Ditches with a length of optical way of 1 cm.
The serum, plasma and urine which are taken away by means of standard methods.
Pancreatic a-amylase in serum or plasma is stable within 1 month at a temperature 2-8ºC. As anticoagulant to use heparin or EDTA.
Pancreatic a-amylase in urine is stable within 1 month at a temperature 2-8ºC provided that at storage of pH it is approximately equal to 7. Before definition to centrifuge the distilled water.