Amylase

Alpha Amylase
5+1 liquid
Cat.No Package Size REFERENCE VALUES ( 37°C)

Serum/plasma Spontaneous Collected
AM260S06 2 x 50 mL 1 x 20 mL urine urine
AM460S55 4 x 50 mL 2 x 20 mL 28-100 U/l <450 U/l <410 u/24h
AM2150S128 2 x 125 mL 1 x 50 ml 0,5-1.67 µkat/l <7,5 µkat/l <6.9 µkat/l/24h
AM240H105Hit I 4 x 50 mL 2 x 20 mL
ASSAY PROCEDURE:
R1 and R2 are ready for use.
METHOD Before use keep at constant temperature of 37°C
Colorimetric Kinetic, EPS/IFCC
Wavelength : Hg 405 nm
Light path: 1 cm
PRINCIPLE: Temperature: 37°C
Enzymatic color test with 4,6 -Ethyliden(G7)-p-nitrophenyl-
Measuring against water (increase of absorbance)
maltoheptaosid (EPS) as substrate.
α-Amylase
5 Ethyliden-G7PNP + 5 H2O → 2 Ethyliden-G5 +2 G2PNP Reaction mixture (only for start with sample):
+ 2 Ethyliden-G4 +2 G3PNP Mix buffer R1 (5 volumes) with start reagent R2 (1
+ Ethyliden-G3 + G4PNP volume)
α-Glucosidase Stability: 2° - 8°C: 20 days
2 G PNP + 2 G PNP + 10H2O → 4 PNP + 10 Glucose
2 3 18° - 22°C: 5 days
(PNP=p-Nitrophenol)
REAGENTS (concentrations in the test): Pipette into cuvettes:

R1: Good's Buffer (pH 7,1) 0,1 mol/l
NaCl 50 mmol/l Start with R2 Serum Urine
MgCl2 10 mmol/l R1 (Buffer) 1000 µl 1000 µl
α-Glucosidase ≥ 2 kU/l Sample 20 µl 10 µl
R2: Good's Buffer (pH 7,1) 0,1 mol/l R2 (Starter Rgt) 200 µl 200 µl
EPS-G7 1,6 mmol/l
The sealed reagents are stable up to the indicated expiry date if Start with sample
stored at +2° - +8°C. Serum Urine
Reaction Mixture 1000 µl 1000 µl
NOTE: 20 µl 10 µl
-Do not pipette by mouth. Avoid contact of the
reagent with skin, as saliva and sweat contain Mix and incubate for 2 min at 37°C. Determine
a-amylase. average from the absorbance difference per
-Reagents contain < 0.95 % sodium azide as a minute after 3 min.
preservative. Avoid swallowing as well as contact
with skin or mucuous membranes. CALCULATION:
p-Nitrophenol is formed by the reaction. This is
dangerous to health when being inhaled or Start with R2:
swallowed at skin contact. In case of skin contact Serum/plasma: U/L= 6157 x ∆A /min
immediately wash off with plenty of water µKat/L= 102,7 x ∆A /min
Urine: U/L= 12218 x ∆A /min
SAMPLE MATERIAL:
µKat/L= 203,7 x ∆A /min
Serum, Heparin- and EDTA- plasma, urine
Start with sample
Serum/plasma: U/L= 5148 x ∆A /min
SAMPLE PREPARATION:
µKat/L= 86 x ∆A /min
No significant activity decrease in serum within
Urine: U/L= 10198 x ∆A /min
5 days.
Stability in urine:: 10 days at + 4°C µKat/L= 170 x ∆A /min
2 days at +25°C .
L-S 04/2009 page 1/2
CALIBRATORS & CONTROLS BIBLIOGRAPHY
For the calibration of automated analyzers Inmesco 1. Lorentz K. α-Amylase. In: Thomas st
L, editor.
Introcal Multicalibrator is recommended, for quality Clinical laboratory diagnostics. 1 ed. Frankfurt:
control use Inmesco normal and abnormal control, TH-Books Verlagsgesellschaft;1998.p.192-202.
2. Moss DW, Henderson AR. Digestive enzymes of
Intronorm I and Intronorm II. pancreatic origin. In: Burtis CA, Ashwood ER, rd
. editors. Tietz Textbook of Clinical Chemistry. 3
ed. Philadelphia: W.B Saunders
PERFORMANCE DATA (37°C) Company;1999.p.689-98.
3. Kruse-Jarres JD, Kaiser C, Hafkenscheid JC,
-Linearity Hohenwallner W, Stein W., Bohner J et al.
Evaluation of a new alpha-amylase assay using
The limit of dilution is found at the 4,6-ethylidene-(G7)-1-4-nitrophenyl-(G1)-
absorbance differences δ A/min = 0.35 based alpha,D-maltoheptaoside as substrate. J Clin
on a measuring time of 3 minutes .At higher Chem Biochem 1989;27:103-13.
4. Hohenwallner W, Stein W, Hafkenscheid JC,
activities mix 0.1ml sample with 1.0ml NaCI Kruse-Jarres JD, Kaiser C, Hubbuch A et al.
solution 0.9% and repeat assay, multiply Reference ranges for alpha-amylase in serum
result by 11. and urine with 4,6-ethylidene-(G7)-1-4-
nitrophenyl-(G1)-alpha,D-maltoheptaoside as
substrate. J Clin Chem Clin Biochem
1989;27:97-101.
- Sensitivity
The sensitivity is equal to 3 U/L
- Precision
Within-run reproducibility
Within series Mean value Standard- CV
n = 20 [U/L] deviation [%] SYMBOLS USED
[U/L]
Sample 1 77,7 1,009 1,298
Sample 2 196 1,834 0,935
Sample 3 72,1 1,700 0,934 For in vitro diagnostic medical use
Between-run reproducibility
Day to day Mean value Standard- CV
n = 20 [U/L] deviation [%]
Batch Code
[U/L]
Sample 1 79,5 0,934 1,176
Sample 2 195 0,674 0,345 Use by
Sample 3 72,5 0,934 1,288
- Correlation Temperature limitation

A comparative study has been performed between
Inmesco method and another commercial reagent
on 36 human serum samples. The parameters of
linear regression are as follows:
Linear regression: y = 1,000 x + 2,480 U/L

Correlation coefficient r = 0.977
INTERFERENCES
- Ascorbic Acid: no interference up to 30 mg/dL

- Bilirubin: no interference up to 40 mg/dL
- Hemoglobin: no interferences up to 250 mg/dL
- Triglycerides: no interference up to 1000 mg/dL
L-S 04/2009 page 2/2

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Alpha Amylase

Cat.No Package Size REFERENCE VALUES ( 37°C)

REAGENTS (concentrations in the test): Pipette into cuvettes:

- Correlation Temperature limitation

Linear regression: y = 1,000 x + 2,480 U/L

- Ascorbic Acid: no interference up to 30 mg/dL

L-S 04/2009 page 2/2

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