Reactivo de amilasa

1. Siemens Healthcare Diagnostics Inc.
© 2008 Siemens Healthcare Diagnostics. All rights reserved.
Siemens Healthcare Diagnostics Inc., Tarrytown, NY 10591-5097 USA
10493947_EN Rev. E, 2011-02 1 - English
Instructions for Use
2011-02
Amylase (AMYLAS)
Method Summary
System New Information
ADVIA® 1200 Updates: New document number; Symbols page added
ADVIA 1650/1800 Updates: New document number; Symbols page added
ADVIA 2400 Updates: New document number; Symbols page added
Item Description
Method Principle Ethylidene Blocked-pNPG7
Specimen Type Human serum, plasma (lithium heparin), and urine
On-board Stability ADVIA 1200: 30 days
ADVIA 1650/1800: 50 days
ADVIA 2400: 50 days
Reagent Storage
Temperature
2–8°C
Calibration Frequency Daily
Reagent Blank (RBL)
Frequency
Daily
Reaction Type Rate (RRA)
Measurement
Wavelength
410/694 nm
Standardization IFCC reference method
Analytical Range 0–1500 U/L
Expected Values Serum:
Urine:
30–118 U/L
≤ 650 U/L
Reagent Code 74042
Calibrator Fixed System Factor Value

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Intended Use
For in vitro diagnostic use in the quantitative determination of amylase activity in human
serum, plasma, and urine on ADVIA Chemistry systems. Such measurements are used
primarily in the diagnosis and monitoring of acute pancreatitis (inflammation of the
pancreas).
Summary and Explanation
The ADVIA Chemistry Amylase (AMYLAS) method is based on the procedure of
Jensen and Wydeveld.1
Principles of the Procedure
The ADVIA Chemistry AMYLAS method uses ethylidene blocked
p-nitrophenyl-maltoheptaoside as substrate. The indicator enzyme α-glucosidase,
used to release the p-nitrophenol, is also employed in the method. The terminal
glucose of the substrate is chemically blocked preventing cleavage by the indicator
enzymes. The released p-nitrophenol is measured at 410/694 nm.
Reaction Equation
Reagents
The reagents are packaged as listed below. Components of the package are available
as a kit only.
Safety data sheets (MSDS/SDS) available on www.siemens.com/diagnostics.
Ed-G7PNP Ed-Gn + Gn-PNP
Gn-PNP PNP + Glucose
REF
Container Size Symbol Contents Amount (mL) No. of Tests
03031177 Amylase Reagents 7 x 355
40-mL Reagent 1 7 x 38
20-mL Reagent 2 7 x 11.7
07498401 Amylase Reagents 7 x 175
20-mL Reagent 1 7 x 18
20-mL Reagent 2 7 x 6.2

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Components and Concentrations
NOTE: Sodium azide can react with copper and lead plumbing to form explosive metal
azides. If disposal into a drain is in compliance with federal, state, and local
requirements, flush reagents with a large amount of water to prevent the buildup of
azides.
For in vitro diagnostic use.
Reagent Preparation and Use
Reagents are ready to use. Before use, gently swirl the reagent to disrupt bubbles and
assure homogeneity.
On-board Reagent Stability (OBS)
For all systems, unopened reagents are stable until the expiration date printed on the
product label when stored at 2° to 8°C. Do not freeze reagents.
For additional details, refer to the Methods Introduction section of the system-specific
Operator’s Guide.
Sample Handling
Siemens Healthcare Diagnostics recommends using serum, plasma (lithium heparin),
or urine for this method. For additional details, refer to Sample Collection and
Preparation in the Methods Introduction section of the system-specific Operator’s
Guide.
For instructions on how to load reagents and run samples, refer to the Daily Operations
section of the system-specific Operator’s Guide.
Reagent Component Concentration
Reagent 1 α-Glucosidase
Sodium azide
≥ 4 KU/L
0.04%
Reagent 2 Ethylidene-4-NP-G7
Sodium azide
22 mmol/L
0.04%
System Stability
ADVIA 1200 30 days
ADVIA 1650/1800 50 days
ADVIA 2400 50 days

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Materials Required but not Provided
The following list contains the materials required, but not provided, to perform this
method:
• sample containers
• system solutions
• control materials (refer to the Quality Control section)*
• reagent container adapters:
• 20-mL adapter (REF 02404085; PN 094-0159-01) for 40-mL slot
(ADVIA 1200, ADVIA 1800)
• 20-mL adapter (REF 05249323; PN 073-0936-01) for 70-mL slot (ADVIA 1200)
• 20-mL adapter (REF 00771668; PN 073-0345-02) for 70-mL slot
(ADVIA 1650, ADVIA 2400)
• 40-mL adapter (REF 08163594; PN 073-0788-01) for 70-mL slot
(ADVIA 1650, ADVIA 2400)
*For storage and stability information, refer to the package insert.
Calibration
Calibration of this method uses a fixed system Factor Value (FV), which is based on the
established molar extinction coefficient of p-nitrophenol at 410 nm, adjusted by patient
sample correlation to the IFCC reference method. One unit of amylase activity is
defined as that amount of enzyme that catalyzes the production of 1 µmol of
p-nitrophenol per minute under the conditions of the method.
For setup and use instructions, refer to the Calibration Overview section of the
system-specific Operator’s Guide.
Reagent Blank (RBL) Frequency
Siemens recommends daily Reagent Blank (RBL) measurement.
Quality Control
Follow government regulations or accreditation requirements for quality control
frequency.
Siemens recommends the use of quality control material from Bio-Rad Laboratories
with at least 2 levels (low and high). A satisfactory level of performance is achieved
when the analyte values obtained are within the Acceptable Control Range for the
system or within your range, as determined by an appropriate internal laboratory
quality control scheme.
The actual frequency of control in a laboratory is based on many factors, such as
workflow, system experience, and government regulation. Each laboratory should
evaluate the controls based on the frequency established by their laboratory
guidelines. When the method is performed, analyze at least 2 levels of controls daily.

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Also, assay controls under the following conditions:
• whenever you use a new reagent lot
• following the performance of any system maintenance, cleaning, or troubleshooting
procedure
• after performing a new calibration
For more information, refer to the Quality Control Overview section of the
system-specific Operator’s Guide.
Limitations of the Procedure
A number of substances cause physiological changes in serum, plasma, or urine
analyte concentrations. A comprehensive discussion of possible interfering
substances, their serum, plasma, or urine concentrations, and their possible
physiological involvements is beyond the scope of this document. Consult listed
references for specific details on known potential interfering substances.2
As with any chemical reaction, you must be alert to the possible effect on results of
unknown interferences from medications or endogenous substances. The laboratory
and physician must evaluate all patient results in light of the total clinical status of the
patient.
Interferences
Siemens tested the following potential interferents and found the results shown below:
ADVIA 1200
Interferent Interferent Level
Amylase Sample
Concentration Interference*
Bilirubin 25.0
(428
mg/dL
µmol/L)
71 U/L NSI
Hemolysis
(hemoglobin)
500
(5.0
mg/dL
g/L)
72 U/L NSI
Lipemia
(from Intralipid)
500
(5.7
mg/dL
mmol/L)**
69 U/L NSI
*NSI = No Significant Interference. A percentage effect ≥ 10% is considered a significant
interference.
**as triolein

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Performance Characteristics
Precision
Each sample was assayed 2 times per run, 1 or 2 runs per day, for at least 20 days.
Precision estimates were computed according to CLSI document EP5-A2, Evaluation
of Precision Performance of Quantitative Measurement Methods; Approved Guideline.3
Data contained in this section represents typical performance for
ADVIA Chemistry systems. Your laboratory data may differ from these values.
ADVIA 1650/1800
Interferent Interferent Level
Amylase Sample
Concentration Interference*
Bilirubin 30
(513
mg/dL
µmol/L)
55 U/L NSI
Hemolysis
(hemoglobin)
525
(5.3
mg/dL
g/L)
55 U/L NSI
Lipemia
(from Intralipid)
650
(7.4
mg/dL
mmol/L)**
55 U/L NSI
*NSI = No Significant Interference. A percentage effect ≥ 10% is considered a significant
interference.
**as triolein
ADVIA 2400
Interferent Interferent Level
Amylase Sample
Concentration Interference*
Bilirubin 25
(428
mg/dL
µmol/L)
149 U/L NSI
Hemolysis
(hemoglobin)
500
(5.0
mg/dL
g/L)
144 U/L NSI
Lipemia
(from Intralipid)
500
(5.7
mg/dL
mmol/L)**
148 U/L NSI
*NSI = No Significant Interference. A percentage effect ≥ 10% is considered a significant
interference.
** as triolein
ADVIA 1200
Within-Run Total
Specimen Type Level (U/L) SD CV (%) SD CV (%)
Serum 63 0.5 0.8 0.9 1.4
Serum 264 1.5 0.6 2.6 1.0
Serum 345 3.1 0.9 4.1 1.2
Urine 71 0.8 1.1 1.6 2.3
Urine 170 1.8 1.0 2.5 1.5

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Analytical Range
The ADVIA Chemistry AMYLAS method is linear from 0 to 1500 U/L for serum, plasma,
and urine.
Siemens has validated an automatic rerun condition for this method that extends the
reportable range up to 4500 U/L for serum and plasma.
Expected Values
The expected values of the ADVIA Chemistry AMYLAS method are 30 to 118 U/L for
serum and plasma, 4 and ≤ 650 U/L for urine.5 Siemens provides this information for
reference. Each laboratory should establish its own normal range. You can enter
normal range values and abnormal range values in the Analytical Parameters
(Chemistry) window.
System Correlation
The performance of the ADVIA Chemistry AMYLAS method (y) was compared with the
performance of the same method on the comparison system (x).
ADVIA 1650/1800
Within-Run Total
Specimen Type Level (U/L) SD CV (%) SD CV (%)
Serum 60 0.6 1.0 0.7 1.2
Serum 242 1.8 0.7 3.1 1.3
Serum 322 2.0 0.6 3.8 1.2
Urine 792 6.1 0.8 57.3 7.2
ADVIA 2400
Within-Run Total
Specimen Type Level (U/L) SD CV (%) SD CV (%)
Serum 53 0.5 0.9 0.6 1.2
Serum 249 1.2 0.5 1.6 0.7
Serum 348 1.7 0.5 1.7 0.5
Urine 55 3.1 5.7 7.3 13.4
Urine 118 0.8 0.7 2.4 2.0
ADVIA 1200
Specimen Type Comparison System (x) N Regression Equation Sy.x r Sample Range
Serum ADVIA 1650 276 y = 1.04x - 1.5 2.9 1.000 7–1315 U/L
Plasma* ADVIA 1200 serum 45 y = 1.01x - 1.0 0.8 1.000 28–287 U/L
Urine ADVIA 1650 63 y = 0.97x + 6.6 10.3 0.999 15–896 U/L
Serum Reference Method 47 y = 0.98x + 6.5 9.7 0.999 39–682 U/L
*lithium heparin

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Standardization
The ADVIA Chemistry AMYLAS method is traceable to the IFCC reference method via
patient sample correlation. See the correlation data in the System Correlation section
for the relationship.
Bibliography
1. Jensen AP, Wydeveld A. α-(p-nitrophenyl) malto hexaoside as a substrate for the
assay of amylase. Nature. 1958;182:525-526.
2. Young DS. Effects of Drugs on Clinical Laboratory Tests. 3rd ed. Washington:
AACC Press; 1990.
3. Clinical and Laboratory Standards Institute (formerly NCCLS). Evaluation of
Precision Performance of Quantitative Measurement Methods; Approved Guideline
- Second Edition. Wayne, PA: Clinical and Laboratory Standards Institute; 2004.
NCCLS Document EP5-A2.
4. Data on file.
5. Tietz NW. Clinical Guide to Laboratory Tests. 2nd ed. Philadelphia, PA: WB
Saunders Company; 1995:52.
Technical Assistance
For customer support, please contact your local technical support provider or
distributor.
www.siemens.com/diagnostics
ADVIA 1650/1800
Specimen Type Comparison System (x) N Regression Equation Sy.x r Sample Range
Serum* Synchron CX4 102 y = 0.72x + 1.9 3.5 0.998 6–549 U/L
Plasma** ADVIA 1650 serum 53 y = 1.08x + 1.5 3.9 0.985 18–110 U/L
Urine* Synchron CX7 82 y = 0.75x + 0.7 13.4 0.993 2–667 U/L
Serum Reference Method 65 y = 0.96x + 15.4 15.3 0.999 43–1327 U/L
*The bias in these comparisons is due to the different substrates used with ADVIA and Beckman methods.
**lithium heparin
ADVIA 2400
Specimen Type Comparison System (x) N
Regression
Equation Sy.x r Sample Range
Serum ADVIA 1650 269 y = 0.98x - 1.2 2.6 0.999 9–1122 U/L
Urine ADVIA 1650 70 y = 0.94x + 0.6 2.4 1.000 0–598 U/L
Serum Reference Method 65 y = 0.99x + 3.6 10.0 0.999 43–1327 U/L

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Trademarks
ADVIA IMS and ADVIA are trademarks of Siemens Healthcare Diagnostics.
Bio-Rad is a trademark of Bio-Rad Laboratories.
Intralipid is a trademark of Fresenius Kabi AB.
Synchron CX4 and CX7 are trademarks of Beckman Coulter, Inc.

10. Amylase (AMYLAS)
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Understanding the Symbols
The following symbols may appear on the product labeling:
Symbol Definition Symbol Definition
In vitro diagnostic medical device Catalog number
Manufacturer
Authorized Representative in the European
Community
CE Mark
CE Mark with identification number of notified
body
Consult instructions for use Caution! Potential Biohazard
Do not freeze (> 0°C) Temperature limitation (2–8°C)
Lower limit of temperature (≥ 2°C) Upper limit of temperature (≤ -10°C)
Keep away from sunlight Use by
Store upright Contains sufficient for (n) tests
Batch code Printed with soy ink
2010-01 Date format (year-month) Recycle
Green dot