Amylase Reagents

Amylase Reagents is an in-vitro reagent kit for the quantitative determination of α-amylase in human serum. This amylase assay kit uses the CNP-G3 method and measures enzyme activity kinetically at 405 nm. It can be used as an amylase test kit in routine clinical chemistry workflows and functions as an amylase kit for semi-automatic and automated analyser applications. 

Pack size: 1 × 10 ml. 

Shelf life: 18 months. 

Storage: 2° to 8°C. 

Product Overview

This amylase reagents kit is based on the 2-chloro-4-nitrophenyl–α-D maltotrioside substrate method. Under the catalytic action of α-amylase, the chromogen CNP is released and measured kinetically. The intensity of CNP formed is directly proportional to α-amylase activity and is measured at 405 nm. 

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Intended Use

This reagent kit is used for in-vitro quantitative determination of α-amylase in human serum. 

Assay Principle

Defined oligosaccharides such as 2-chloro-4-nitrophenyl–α-D maltotrioside are cleaved under the catalytic action of α-amylase. The release of the chromogen CNP is then measured. The intensity of chromogen formed is directly proportional to α-amylase activity and is measured kinetically at 405 nm. 

Reaction

2-chloro-4-nitrophenyl–α-D Maltotrioside —α-amylase→ chloro-nitro phenol (CNP) + CNP-G2 + 9G3 + G. 

Kit Contents

• Reagent 1: Amylase Enzyme Reagent
• Product Insert: 01 No. 

Working Reagent Preparation

α-Amylase liquid reagents are ready to use. 

Storage and Stability

The reagent should be stored at 2–8°C and is stable till the expiry date mentioned on the labels. Do not freeze the reagent. 

Specimen Collection and Storage

Unhemolysed serum is the specimen of choice. E.D.T.A., oxalate or citrate inhibit amylase activity and cannot be used. Amylase in serum is reported to be stable for one week at room temperature and two months when stored at 2–8°C. 

Precautions

• For in-vitro diagnostics use only
• Do not inhale the reagent or pipette by mouth
• Do not freeze or expose the reagents to higher temperature
• Bring all reagents to room temperature before the assay
• Avoid contamination during assay process
• Use clean glassware free from dust or debris
• Do not use the reagent if it is hazy or cloudy 

Test Procedure

Manual Procedure

Pipette into test tubes:

• Amylase Enzyme Reagent: 1.0 ml
• Sample: 25 µl

Mix thoroughly and transfer the assay mixture immediately to the thermostated cuvette. Record the first absorbance reading after 60 seconds and then take two more absorbance readings with 60 second intervals at 405 nm. 

Automated Procedure

Refer to specific instrument application instructions. 

Calculations

Calculate the average change in absorbance per minute (ΔAbs/minute).

Activity of α-Amylase in IU/l = ΔAbs/minute × 3178. 

Input Parameters

• Type of reaction: Kinetic
• Slope of reaction: Increasing
• Wavelength: 405 nm
• Factor: 3178
• Incubation time: 60 sec
• Interval time: 60 sec
• Interval No.: 2
• Units: IU/l
• Temperature: 37°C
• Upper normal value: 140 IU/l
• Lower normal value: 25 IU/l
• Linearity: 2000 IU/l
• Enzyme reagent volume: 1.0 ml
• Sample volume: 25 µl 

Normal Values

Serum: 25–140 IU/l

It is recommended that each laboratory should establish its own normal range. 

Performance Characteristics

• Linearity: 2000 IU/l
• Comparison: r = 0.98; y = 0.97x + 4.4
• Precision: within-run and run-to-run precision values are listed in the IFU table
• Specificity: no interference with 1 gm/l haemoglobin and bilirubin 30 mg/dl 

Clinical Significance

The IFU states that α-amylase determinations are of importance in pancreatic diagnostics and are mainly used in the diagnosis and monitoring of acute pancreatitis. 

Automated Applications

α-Amylase liquid reagents can be used with Hitachi 700 series, RA50, 1000 XT, Express 550, Syncron CX4, LISA 200, BTR 810/820/830, Erbachem-5, Ranlab and similar systems. Application sheets for specific semi-automatic and batch analysers are available on request. 

Quality Control

For accuracy, it is necessary to run known serum controls with each assay. 

FAQs

What is this amylase reagents kit used for?

It is used for the in-vitro quantitative determination of α-amylase in human serum. 

Is this an amylase assay kit?

Yes. This amylase assay kit uses the CNP-G3 method and measures α-amylase activity kinetically at 405 nm. 

Can this be used as an amylase test kit in routine labs?

Yes. The IFU supports use in routine clinical chemistry workflows, including semi-automatic and automated analyser applications. 

What specimen type should be used?

Unhemolysed serum is the specimen of choice. EDTA, oxalate and citrate should not be used because they inhibit amylase activity. 

What is the wavelength used in this amylase kit?

The assay is measured at 405 nm. 

What is the stated linearity of the kit?

The IFU states a linearity of 2000 IU/l. 

How is α-amylase activity calculated?

Activity of α-Amylase in IU/l = ΔAbs/minute × 3178