1.alpha Amylase

AlphaAMYLASE
Kinetic method for the quantitative determination of

Alpha Amylase in serum, plasma and urine
ORDER INFORMATION ANALYTICAL PROCEDURE

REF Kit size Working temperature 37 °C
GA4175 00 5x20 ml Wavelength 405 nm (400-410 nm)
KL4175 00 12x20 ml Optical path 1 cm
BK4175 00 2x40 ml Reaction Kinetic (increase)
INDICATION Allow the reagents to reach working temperature before

-Amylase is an enzyme that is secreted by the salivary and using.
pancreatic glands. It is important for the digestion of
starches and is rapidly cleared by the kidneys. Pipette into disposable or well clean cuvettes:
Increased values of Amylase are found in acute Reagent Blank Sample
pancreatitis, obstruction of the pancreatic ducts, and Reagent A 1000 l 1000 l
(mildly) in obstruction of the parotid gland. Decreased Distilled H2O 25 l -
values are found in acute or chronic hepatocellular damage. Sample - 25 l
Mix and incubate for 1 minute at 37 °C.
METHOD PRINCIPLE Read initial absorbance and repeat the absorbance
-Amylase catalyzes hydrolysis of 2-chloro-4-nitrophenyl-- reading after 1, 2, 3 minutes against reagent blank.
maltotrioside (CNP-G3) to glucose polymers and p- Calculate ∆A/min.
nitrophenyl oligosaccharide at short chain producing 2-
chloro-4-nitrophenol (CNP).
CALCULATION OF RESULTS
The increased extinction can be measured by
spectrophotometry at 405 nm and results proportional to
Activity (U/l) = A/min x 3178
the activity of - Amylase in the sample .
COMPOSITION
REFERENCE VALUES
REAGENT A:
CNP-G3 2 mmol/l
Serum or plasma up to 90 U/l
Sodium chloride 250 mmol/l
Urine up to 480 U/l
Calcium chloride 6 mmol/l
MES buffer 100 mmol/l
Each laboratory should establish reference ranges for its
Potassium thiocyanate 600 mmol/l
own patients population.
Sodium azide < 0.1%
Preparation:
ANALYTICAL PERFORMANCES
The reagent is liquid ready to use. Precision
Within-run and between-run coefficients of variation have
Storage and stability been calculated on replicates of three samples at different
Store at 2-8 °C. Do not freeze the reagents! The reagents enzymatic activities. The obtained results are reported in
are stable until the expiry date, if contamination and the following tables:
evaporation are avoided, protected from light. The above
conditions are valid if the vials are opened just only for the Within Run Between Run
time to take the reagent, closed immediately with their cap Mean
Sample SD %CV SD %CV
and stored at the indicated conservation temperature. (U/l)
Serum 1 72.50 0.92 1.3 1.74 2.4
Saliva and skin contain Amylase: never pipette by mouth
Serum 2 124.80 1.19 1.0 4.94 4.0
and avoid skin contact with reagents.
Serum 3 175.32 1.74 1.0 6.13 3.5
ANCILLARY EQUIPMENT
 Automatic pipettes Linearity
 Photometer The assay is linear up to 1500 U/l.
 Analysis cuvettes (optical path = 1 cm)
 Temperature controlled water bath Sensitivity
 NaCl solution 9 g/l Test sensitivity, in terms of limit of detection, is 3 U/l.
SAMPLES Correlation
Serum, heparin plasma and urine. A correlation study comparing the present method an a
Stable 7 days at 2-8 °C or 30 days at -20 °C. commercial one gave the following results:
Specimen collection / Preanalytical factors y = 1.145x + 1.600 U/l r = 0.9967

It is recommended that specimen collection should be
carried out in accordance with NCCLS Document H11-A3. Interferences
Hemoglobin > 500 mg/dl
INTERNAL QUALITY CONTROL Bilirubin > 40 mg/dl
It is recommended to use commercial Quality Control sera Triglycerides > 1500 mg/dl
with known -Amylase activity. Check that the values
obtained are within the reference range provided.
Pag. 1/2
GA4175 00 - KL4175 00 - BK4175 00 Alpha AMYLASE
PRECAUTIONS IN USE
The Reagent A (REF GB017500) is not considered
harmful (GHS00) according to (CE) n. 1272/2008
directive about classification, packaging and labelling of
dangerous substances.
Reagent A:
Supplemental Hazard statement Codes:
EUH032 - Contact with acids liberates very toxic gas.
EUH210 - Safety data sheet available on request.
However, the reagents should be handled with caution,
avoiding swallowing and contact with skin, eyes and
mucous membranes.
The use of laboratory reagents according to good laboratory
practice is recommended.
Waste Management
Please refer to local legal requirements.
BIBLIOGRAPHY
1. Lorentz K. -Amylase. In: Thomas L, editor. Clinical
laboratory diagnostics. 1st ed. Frankfurt: TH-Books
Verlagsgesellschaft; 1998, p. 192-202.
2. Tietz N W (ed): Clinical guide to laboratory tests, 3rd
reprint. Philadelphia, PA: WB Saunders Company;
1995:46-49.
3. Jakobs DS, Kasten Jr. BL, Demmott WR, Wolfson WL:
“Laboratory Test Handbook”, Lexi-Comp and Williams &
Wilkins Ed. 2nd Edition (1990).
4. International Federation of Clinical Chemistry and
Laboratory Medicine (IFCC): “Approved
Recommendation on IFCC Methods for the
measurement of catalytic concentration of enzymes –
Part 9. IFCC Method for -Amylase (1,4--D-Glucan 4-
Glucanohydrolase, EC 3.2.1.1)” – Clin Chem Lab Med
1998; 36(3):185-203.
5. EU-Dir 1999/11 Commission Directive of 8 March 1999
adapting to technical progress the principles of good
laboratory practice as specified in Council Directive
87/18/EEC.
6. NCCLS Document, “Procedures for the collection of
arterial blood specimens”, Approved Standard, 3rd Ed.
(1999).
NOTE: Changes Highlighted
Pag. 2/2
AMS Srl. – Registered Office and Plant

Via E. Barsanti 17/A Met. GA417500.3 eng
00012 Guidonia (Rome) – Italy Edition: 2018/11
Ph. + 39 0774 354441
Fax + 39 0774 578035
www.amsalliance.com - [email protected]

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AlphaAMYLASE

Kinetic method for the quantitative determination of

ORDER INFORMATION ANALYTICAL PROCEDURE

INDICATION Allow the reagents to reach working temperature before

Specimen collection / Preanalytical factors y = 1.145x + 1.600 U/l r = 0.9967

NOTE: Changes Highlighted

AMS Srl. – Registered Office and Plant

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