OSR61154 Infinity™ Ammonia Reagent

2 x 16 mL For beckman Coulter AU Chemistry Analyzers

®

1 x 3 mL
INTENDED USE REAGENT PREPARATION
Reagent for the quantitative determination of Ammonia (NH3) concentrations in Reagent and Standard are provided ready to use
human plasma for use on the Beckman Coulter AU® Chemistry Analyzers.
STAbILITy and STORAGE
SUMMARy1,2,3 1. The unopened reagent and standard are stable until the expiration date when
Ammonia, derived from the catabolism of amino acids and from the action of stored at 2-8°C.
intestinal bacteria on dietary protein, is converted to urea in the liver hepatocytes 2. Once opened the reagent and standard are stable in the bottles provided until
and so rendered non toxic. Under normal circumstances the concentration of the expiry date stated, provided that it is capped when not in use and stored
ammonia in the circulation remains low, typically less than at 2-8°C. When stored on board, the reagent is stable for 14 days.
50 µmol/L (85 µg/dL). Studies have shown that excess ammonia can have a toxic
effect on the central nervous system and clinical manifestations are typically Indications of Reagent Deterioration
neurological disturbances. Turbidity and/or failure to recover control values within the assigned range.

Elevated levels of ammonia may be either due to: (i) Inborn errors of metabolism; SPECIMEN COLLECTION and PREPARATION1
or (ii) Secondary to other conditions. It is recommended that human plasma be collected in EDTA or heparin
Inborn errors of metabolism are the major cause of elevated ammonia in infants and (not ammonium heparin). Ideally, the collection tube should be completely filled
usually the result of urea cycle enzyme deficiencies. Inherited disorders affecting with blood and immediately placed on ice. Centrifuge (cold) the sample as soon as
the metabolism of the dibasic amino acids (lysine and ornithine) and those involving possible and separate plasma and store at 2-4°C until analysis.
the metabolism of organic acids may also produce elevated levels of circulating
ammonia. Elevated ammonia may also be observed in severe liver failure as may Sample Storage and Stability
occur in Reye’s Syndrome, viral hepatitis or cirrhosis. Ammonia samples are stable for 3 hours at 2-4°C or 24 hours at –20°C.1

METHODOLOGy1 LIMITATIONS
A number of methods have been developed for the estimation of plasma ammonia Interfering Substances
and these can be broadly classified into either indirect or direct methods. In the 1. Hemolyzed samples should not be used as erythrocytes contain levels of
indirect procedures, ammonia is first of all isolated, for example by the addition ammonia approximately 3 times that of plasma.1
of alkali or the use of a cation exchange resin, after which it is measured 2. No interference from pyruvate was observed up to a level of 0.01 mg/dL (0.75
colourimetrically by nesslerization or Berthelot reaction. These procedures are mmol/L).
not easily automated or require dedicated equipment. Direct procedures, such as 3. No interference from ALT was observed up to a level of 2400 U/L.
enzymatic methods, are more widely used in routine laboratories as they do not 4. Bilirubin: No significant interference up to 1.8 mg/dL unconjugated Bilirubin.
require the separation of ammonia from the specimen prior to the analytical step. 5. Lipemia: No significant interference up to 100 mg/dL triglycerides.
Direct procedures are therefore more easily automated. The Infinity™ ammonia 6. Reliable estimations of ammonia can only be achieved if steps are taken to
reagent is a direct enzymatic procedure based on the following reaction sequence: avoid contamination from ammonia. Sources of contamination include, but are
not restricted to, cigarette smoking (patient and collection staff), laboratory
Glutamate dehydrogenase (GLDH) atmosphere, laboratory glassware or other reagents on the carousel which
NH4 + α ketoglutarate + NADH ——————————————> Glutamate + NAD + H2O
contain Ammonia. In the case of the latter, avoid use of the ammonia containing
The reagent contains LDH in excess, to rapidly reduce endogenous pyruvate so reagents together with OSR61154 to mitigate against atmospheric ammonia
that it does not interfere with the assay system. The Beckman Coulter Ammonia transfer. Contact your local Beckman Coulter representative for further
reagent also incorporates a patented stabilization process which renders the reagent information.
stable in the liquid phase.
Dynamic Range
REAGENT and STANDARD COMPOSITION Beckman Coulter ammonia procedure is linear from 10 to 600 µmol/L (17 - 1020
Ammonia Reagent µg/dL).
α-Ketoglutarate 7.5 mmol/L Specimens with Ammonia concentrations greater than 600 µmol/L (1020 µg/dL)
NADH >0.2 mmol/L should be diluted with ammonia free water and reassayed. Multiply results by the
GLDH (Micro-organism) >4,000 U/L dilution factor.
LDH (Micro-organism) >30,000 U/L
Tris Buffer 100 mmol/L ASSAy PROCEDURE
Preservative Materials Provided
• Infinity™ Ammonia Reagent
Ammonia Standard • Infinity™ Ammonia Standard
Ammonium salt 59 µmol/L (100 µg/dL) Suggested Anaylical Parameters
Refer to the User Guide Accompanying the instrument
WARNING AND PRECAUTIONS
1. For in vitro diagnostic use only. Do not ingest. Harmful if swallowed. Avoid contact Calibration
with skin and eyes. If spilled, thoroughly wash affected areas with water. The calibration frequency for this procedure is 7 days. Calibration of this ammonia
2. Contains sodium azide (0.1% W/V). Sodium azide preservative in diagnostic procedure is accomplished by use of the Infinity™ Ammonia Standard provided
reagents may react with lead joints in copper drain lines to form explosive in the kit. The Standard has been manufactured gravimetrically using a material
compounds. Even though the reagent contains minute quantities of sodium traceable to an in-house certified material.
azide, drains should be well flushed with water when discarding the reagent.
For further information consult the Safety Data Sheet. Recalibration of this procedure is required when a reagent lot number has changed
3. This product contains animal source material. Handle and dispose of this product or there is an observed shift in control values, if a critical part of the analyzer is
as if it were potentially infectious. replaced or, if a major preventative maintenance procedure was performed on
the analyzer.
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Ammonia
Quality Control LOD = LOB + 2SDWR
During operation of the Beckman Coulter AU analyzer at least two levels of an LOB = Limit of Blank
appropriate quality control material should be tested a minimum of once a day. In SDWR = Within Run standard deviation of a low level sample
addition, controls should be performed after calibration, with each new lot of reagent, When run as recommended the lowest limit of detection is 7.1 μmol/L.
and after specific maintenance or troubleshooting steps described in the appropriate
Beckman Coulter AU User’s Guide. Quality control testing should be performed in REFERENCES
accordance with regulatory requirements and each laboratory’s standard procedure. 1. Clinical Chemistry Infobase: A Scientifi c & Management Cyclopedia. Pesce-
Kaplan Publishers 1996; 2246-2320.
Results
Results in µmol/L will be automatically printed for each sample assayed. To work 2. Tietz Textbook of Clinical Chemistry. Burtis CA and Ashwood ER (Eds). Second
in (µg/dL), the result must be multiplied by 1.7 Edition, WB Saunders Company, 1994; 32:1485-88.

EXPECTED VALUES5 3. The Diagnosis of Urea Cycle Disorders, Lab Medica International, May/June
16 - 53 µmol/L (27 - 90 µg/dL) 1993; 13-17.

The quoted values were derived from a normal population and should serve as a 4. Young DS. Effects of Drugs on Clinical Laboratory Tests Third Edition 1990; 3:
guide only. It is recommended that each Laboratory verify this range or derives a 30-2.
reference interval for the population that it serves.
5. Ektachem Multilayer Dry Film Assay for Ammonia Evaluated. J Clin Chem 1985:
Vol 31:12:2012-2014.
SPECIFIC PERFORMANACE CHARACTERISTICS
Precision7 6. Clinical and Laboratory Standards Institute. User evaluation of Precision
Estimates of precision, based on CLSI recommendations6, are less than 5% within Performance of Clinical Laboratory Devices. CLSI:2004, CLSI Publication
run and total precision is less than 5%. Assays of control sera products were EP5-A2.
performed and the data produced following the CLSI guidlines above.
7. Clinical and Laboratory Standards Institute. Protocols for Determination of Limits
N= 20 WITHIN RUN TOTAL of Detection and Limits of Quantitation. CLSI:2012, CLSI publication EP17-A2
Mean (μmol/L) SD CV% SD CV%
42.4 1.772 3.7 2.365 5.0
192 1.867 0.9 5.569 2.8

Method Comparison7,8
A comparison of this Beckman Coulter ammonia method (Method 1) vs an on
market method (Method 2) was run per CLSI EP09-A2 using 61 patient samples.
The resulting data is as follows:
Correlation Coefficient: r = 0.973
Regression equation: Method 1 = 0.903X + 2.42
Range of patients: 100 - 600 μmol/L

Lower Limit of Detection

The lower limit of detection was determined using CLSI EP17-A28 using
the formula:
Where:

Symbols Fisher Diagnostics

Authorized Representative in the European Community a division of Fisher Scientific Company, LLC
a part of Thermo Fisher Scientific Inc.
In vitro Diagnostic Medical Device Middletown, VA 22645-1905 USA

Batch code/Lot number

Catalogue number

Consult instructions For Use WMDE

Bergerweg 18
Reagent 6085 AT Horn
The Netherlands
Standard

Temperature Limitation

Use by/Expiration Date

CAUTION. Consult instructions for use Produced by Fisher Diagnostics for:

Beckman Coulter, Inc.
Manufactured by
205 S Kraemer Blvd
Brea, CA 92821 USA

Esoterics Ammonia_BAOSR6x154_US_04
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