3-Amylase

1. pancreatic amylase-pancreas 2.salivary amylase- salivary glands

a-amylase originates from what organ?

a-amylase

are hydrolytic enzymes which break down starch to maltose

The decrease in intensity of dark blue color at 660 nm. Rate of increasing absorbance at 405 nm: resulting to p-nitrophenol from hydrolysis

mode of measurement in amyloclastic & enzymatic/colorimetric

1cm

optical path

pancreas intestinal tract

pancreatic amylase is produced by __? and released into the ___

kidney urine urinary amylase activity

the amylase present in the body is eliminated through the ____ and excreted in ___ therefore elevation of serum activity is reflected in a rise of ??

U/L or units/100 ml sample U/L or µkat/L

unit of measurement in amyloclastic & enzymatic/colorimetric

200U/L max of 0.35 diluted 1+9 with NaCl multiply x 10

upper limit on automated systems of a-AMY upper limit on manual procedure? -if values exceed, what to do?

660nm Hg 405nm

wavelength used in amyloclastic & enzymatic/colorimetric

against reagent blank

measurement of a-AMY

Anhydrous disodium phosphate (pH 7.0 ± 0.1; Na2HPO4) Good's Buffer (pH - 7.15)

Buffer used & pH in amyloclastic & enzymatic/colorimetric

3U/L

lower limit of a-AMY

Endpoint Spectrophotometry UV, 3 point Kinetic, and Spectrophotometry

category of estimation in amyloclastic & enzymatic/colorimetric

Good's buffer (pH7.15; 0.1mol/L) NaCl (62.5mmol/L) Mg2Cl (12.5mmol/L) α-glucosidase (≥ 2kU/L) Good's buffer (pH7.15; 0.1mol/L) EPS-G7 (8.5mmol/L)

components of reagent 1 & 2 for a-amylase

Starch-iodine complex; Colorless (Initial color is Blue) P-nitrophenol (PNP; yellow if there is an increase in serum amylase activity; colorless if normal)

end product & color in amyloclastic & enzymatic/colorimetric

4554

factor used to calculate A/min of a-amylase

few hours after 12hrs after 5 days

in acute pancreatitis, blood amylase activity increases within how many hours from onset of abdominal pain, peaks and returns to normal

measurement of AMY in serum

it is used for the diagnosis of pancreatic disorders as well as detect devt of complications

-substrate 4,6-ethylidene-(G7)-p-nitrophenyl-(G1)-α-D-maltoheptaoside (EPS-G7) is cleaved by α-amylases into various fragments. -further hydrolyzed in a second step by α-glucosidase producing glucose and p-nitrophenol. -increase in Abs represents = the total (pancreatic and salivary) amylase activity in the sample.

principle of a-amylase determination

7.5 minutes (5 minutes pre-warming - to equilibrate mixture, correct lag phase) 5 minutes (2 minutes initial)

reaction time time in amyloclastic & enzymatic/colorimetric

serum/plasma -male & female= <100 (<1.67) urine -female= <447 (<7.45) -male= <491 (<8.18)

reference range of amylase in both serum/plasma & urine

160 U/100 mL or 111-296 U/L

reference range of amylase in iodometric/amyloclastic method

salivary glands saliva

salivary amylase is synthesized in the_____ and secreted into the _____

Serum (0.1 ml) Serum (0.02 ml or 20 µL)

sample and sample volume used in amyloclastic & enzymatic/colorimetric

ascorbic acid- >30mg/dl bilirubin- >40mg/dl hgb- >550mg/dl lipemia- >1000mg/dl triglycerides

sources of interference

lipase

specificity of a-amylase for pancreatic disorders is not very high -elevated lvls are measured alo in non-pancreatic diseases (parotitis & renal insufficiency) -therefore for confirmation of acute pancreatitis, we need another measurement of ___?

7 days at 20-25°C; 7 days at 4-8°C; 1 year at -20°C 2 days at 20-25°C; 10 days at 4-8°C; 3 weeks at -20°C

stability of a-AMY in serum/plasma stability of a-AMY in urine

6 months at 2-8°C; 4 weeks at 15-25°C

stability of reagent

Lintner's Soluble Starch 4,6-ethylidene-(G7)-p-nitrophenyl-(G1)- α Lintner's Soluble Starch -D-maltoheptaoside (EPS-G7

substrate used in amyloclastic & enzymatic/colorimetric

37C

temperature