ALANINE AMINOTRANSFERASE

en
7D56
304663/R02

ALANINE
AMINOTRANSFERASE
This package insert contains information to run the Alanine Aminotransferase assay on the ARCHITECT c Systems.

Read Highlighted Changes Revised September 2012

Package insert instructions must be carefully followed. Reliability of assay results cannot be guaranteed if there are
any deviations from the instructions in this package insert.

Customer Service: Contact your local representative or find country-specific contact information on
www.abbottdiagnostics.com.

Key to Symbols Used

Authorized Representative in the
Reagent 2
European Community
Identifies products to be used
Catalog number/List number
together
Global Trade Item Number Serial number
Information needed for United States
Consult instructions for use
of America only
In Vitro Diagnostic Medical Device Manufacturer

Batch code/Lot number Sufficient for

PRODUCT OF CANADA Product of Canada Temperature limitation

Reagent 1 Use by/Expiration date

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NAME WARNINGS AND PRECAUTIONS
ALANINE AMINOTRANSFERASE Precautions for Users
•
INTENDED USE • For In Vitro Diagnostic Use.
The Alanine Aminotransferase (ALT) assay is used for the quantitation of
• Do not use components beyond the expiration date.
alanine aminotransferase in human serum or plasma.
• Do not mix materials from different kit lot numbers.
SUMMARY AND EXPLANATION OF TEST • CAUTION: This product requires the handling of human specimens.
It is recommended that all human sourced materials be considered
Alanine Aminotransferase (ALT), also referred to as glutamate pyruvate
potentially infectious and be handled in accordance with the OSHA
transaminase (GPT), is an enzyme involved in amino acid metabolism.
Standard on Bloodborne Pathogens.1 Biosafety Level 22 or other
It is found in many tissues, but the highest levels are found in liver and
appropriate biosafety practices3,4 should be used for materials that
kidney tissues. Tissue destruction leads to the release of the intracellular
contain or are suspected of containing infectious agents.
enzyme into the circulating blood. Markedly elevated serum ALT levels
may be found in a variety of diseases which involve the liver, such as • Safety Data Sheets are available at www.abbottdiagnostics.com or
hepatitis, mononucleosis, and cirrhosis. These very high levels of ALT contact your local representative.
are not usually observed in other disease processes, e.g., myocardial
infarction; thus, ALT is regarded as a reasonably specific indicator of SPECIMEN COLLECTION AND HANDLING
liver disease. Suitable Specimens
Serum and plasma are acceptable specimens.
PRINCIPLES OF PROCEDURE
CAUTION: Erythrocytes contain approximately 3 to 5 times more ALT
ALT present in the sample catalyzes the transfer of the amino group than does serum.5
from L-alanine to α-ketoglutarate, forming pyruvate and L-glutamate.
Pyruvate in the presence of NADH and lactate dehydrogenase (LD) Hemolysis in serum or plasma can increase test results.
is reduced to L-lactate. In this reaction NADH is oxidized to NAD. The • Serum: Use serum collected by standard venipuncture techniques
reaction is monitored by measuring the rate of decrease in absorbance into glass or plastic tubes with or without gel barriers. Ensure
at 340 nm due to the oxidation of NADH to NAD. complete clot formation has taken place prior to centrifugation.
Methodology: NADH (without P-5'-P) Centrifuge according to tube manufacturer’s instructions to ensure
proper separation of serum from blood cells.
REAGENTS Some specimens, especially those from patients receiving
anticoagulant or thrombolytic therapy, may take longer to complete
Reagent Kit their clotting processes. Fibrin clots may subsequently form in these
7D56-21 ALT is supplied as a liquid, ready-to-use, two-reagent kit sera and the clots could cause erroneous test results.
which contains: • Plasma: Use plasma collected by standard venipuncture techniques
10 x 70 mL into glass or plastic tubes. Refer to table below for acceptable
anticoagulants. Ensure centrifugation is adequate to remove platelets.
10 x 21 mL
Centrifuge according to tube manufacturer’s instructions to ensure
Estimated tests per kit 3,621 proper separation of plasma from blood cells.
Calculation is based on the minimum reagent fill volume per kit. For total sample volume requirements, refer to the ASSAY PARAMETERS
section of this package insert and Section 5 of the ARCHITECT System
Reactive Ingredients Concentration Operations Manual.
β-NADH 0.16 mg/mL Analyte Recovery
Lactate dehydrogenase 2.57 U/mL Analyte recovery in serum/plasma specimens was determined as a
L-Alanine 392 mmol/L mean %Recovery of serum collected in glass tubes.
α-Ketoglutaric acid 77 mmol/L
Anticoagulant %Recovery
L-alanine 1,000 mmol/L Lithium heparin (with or without gel barrier) 100.3
Sodium heparin 100.1
REAGENT HANDLING AND STORAGE EDTA 97.1
Reagent Handling Sodium citrate 80.2
Remove air bubbles, if present in the reagent cartridge, with a new Plastic tube/serum 99.9
applicator stick. Alternatively, allow the reagent to sit at the appropriate
storage temperature to allow the bubbles to dissipate. To minimize SST gel tube/serum 100.8
volume depletion, do not use a transfer pipette to remove the bubbles. Do not use ammonium heparin.6
CAUTION: Reagent bubbles may interfere with proper detection of
reagent level in the cartridge, causing insufficient reagent aspiration
Specimen Storage
which could impact results. Serum and Plasma: It is recommended that specimens be assayed
on the day of collection.7,8 Separated specimens are stable for 3 days
Reagent Storage at 30°C, 7 days at 2 to 8°C, or 60 days at -40°C or colder.5,9-17 When
Unopened reagents are stable until the expiration date when stored samples were stored at -20°C for 8 days, an 11% reduction in ALT
at 2 to 8°C. activity was observed; a 20% reduction in ALT activity was observed
Reagent stability is 27 days if the reagent is uncapped and onboard. when specimens were stored at -20°C for 1 month.18
NOTE: Stored specimens must be inspected for particulates. If present,
Indications of Deterioration mix and centrifuge the specimen to remove particulates prior to testing.
Instability or deterioration should be suspected if there are precipitates,
visible signs of leakage, extreme turbidity, microbial growth, if calibration
does not meet the appropriate package insert and/or ARCHITECT
System Operations Manual criteria, or if controls do not meet the
appropriate criteria.

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PROCEDURE EXPECTED VALUES
Materials Provided Reference Range
7D56 ALT Reagent Kit Serum19,20/Plasma
Materials Required but not Provided Range (U/L)
• Control Material Adult 0 to 55
• Saline (0.85% to 0.90% NaCl) for specimens that require dilution
It is recommended that each laboratory determine its own reference
Assay Procedure range based upon its particular locale and population characteristics.
For a detailed description of how to run an assay, refer to Section 5 of
the ARCHITECT System Operations Manual. SPECIFIC PERFORMANCE CHARACTERISTICS
Specimen Dilution Procedures Linearity
The ARCHITECT c Systems have an automatic dilution feature; refer to ALT is linear up to 942 U/L.
Section 2 of the ARCHITECT System Operations Manual for additional Flex Rate Linearity is 4,113 U/L. To use Flex Rate Linearity, the operator
information. must edit the linear high value to 4,113 on the Configure assay
Serum and Plasma: Specimens with ALT values exceeding 942 U/L parameters window, Results view.
(4,113 U/L for Flex Rate Linearity) are flagged and may be diluted by Linearity was verified using Clinical and Laboratory Standards Institute
following either the Automated Dilution Protocol or the Manual Dilution (CLSI) protocol NCCLS EP6-P.21
Procedure.
Limit of Detection (LOD)
Automated Dilution Protocol The LOD for ALT is 2.0 U/L. The LOD is the mean concentration of
If using the Automated Dilution Protocol, the system performs a an analyte-free sample + 2 SD, where SD = the pooled, within run
1:5 dilution of the specimen and automatically corrects the concentration standard deviation of the analyte-free sample. A study performed on an
by multiplying the result by the appropriate dilution factor. ARCHITECT c System produced an LOD for ALT of 1.3 U/L.
Manual Dilution Procedure Limit of Quantitation (LOQ)
Manual dilutions should be performed as follows: The LOQ for ALT is 5.1 U/L. The LOQ is the analyte concentration at
• Use saline (0.85% to 0.90% NaCl) to dilute the sample. which the CV = 20%.
• The operator must enter the dilution factor in the patient or control
order screen. The system uses this dilution factor to automatically
Interfering Substances
correct the concentration by multiplying the result by the entered Interference studies were conducted using CLSI protocol NCCLS
factor. EP7-P.22 Interference effects were assessed by Dose Response and
• If the operator does not enter the dilution factor, the result must Paired Difference methods, at the medical decision level of the analyte.
be multiplied by the appropriate dilution factor before reporting the
result. Interfering Target Observed
Interferent Concentration N
Substance (U/L) (% of Target)
NOTE: If a diluted sample result is flagged indicating it is less than the
linear low limit, do not report the result. Rerun using an appropriate 30 mg/dL (513 μmol/L) 4 53.1 95.3
dilution. For detailed information on ordering dilutions, refer to Section 5 Bilirubin
60 mg/dL (1,026 μmol/L) 4 53.1 88.1
of the ARCHITECT System Operations Manual.
750 mg/dL (7.5 g/L) 4 47.4 107.9
Hemoglobin
CALIBRATION 1,000 mg/dL (10.0 g/L) 4 47.4 111.0
Calibration is stable for approximately 27 days (648 hours) and is
550 mg/dL (5.5 g/L) 4 50.6 97.2
required with each change in reagent lot number. Verify calibration with Intralipid
at least two levels of controls according to the established quality control 625 mg/dL (6.25 g/L) 4 50.6 96.8
requirements for your laboratory. If control results fall outside acceptable
ranges, recalibration may be necessary. Bilirubin solutions at the above concentrations were prepared by
addition of a bilirubin stock to human serum pools. Hemoglobin
A calibration factor must be entered on the Configure assay parameters solutions at the above concentrations were prepared by addition of
window, Calibration view. Refer to the ASSAY PARAMETERS section of hemolysate to human serum pools. Intralipid solutions at the above
this package insert for the specific factor. concentrations were prepared by addition of Intralipid to human serum
For a detailed description of how to calibrate an assay, refer to Section 6 pools.
of the ARCHITECT System Operations Manual. Interferences from medications or endogenous substances may affect
results.23
QUALITY CONTROL
The following is the recommendation of Abbott Laboratories for quality Precision
control. As appropriate, refer to your laboratory standard operating The imprecision of the ALT assay is ≤ 5.2% Total CV. Representative
procedure(s) and/or quality assurance plan for additional quality control data from studies using CLSI protocol NCCLS EP5-A24 are summarized
requirements and potential corrective actions. below.
• Two levels of controls (normal and abnormal) are to be run every
Control Level 1 Level 2
24 hours.
• If more frequent control monitoring is required, follow the established N 80 80
quality control procedures for your laboratory. Mean (U/L) 29.9 82.8
• If quality control results do not meet the acceptance criteria
defined by your laboratory, patient values may be suspect. Follow SD 0.44 0.39
the established quality control procedures for your laboratory. Within Run
%CV 1.5 0.5
Recalibration may be necessary.
• Review quality control results and acceptance criteria following a SD 0.38 0.54
Between Run
change of reagent lot. %CV 1.3 0.7
RESULTS SD 1.43 1.51
Between Day
Refer to Appendix C of the ARCHITECT System Operations Manual for %CV 4.8 1.8
information on results calculations.
SD 1.55 1.65
Representative performance data are given in the EXPECTED VALUES Total
and SPECIFIC PERFORMANCE CHARACTERISTICS sections of this %CV 5.2 2.0
package insert. Results obtained in individual laboratories may vary.

LIMITATIONS OF THE PROCEDURE

Refer to the SPECIMEN COLLECTION AND HANDLING and SPECIFIC
PERFORMANCE CHARACTERISTICS sections of this package insert.

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SPECIFIC PERFORMANCE CHARACTERISTICS BIBLIOGRAPHY (Continued)
(Continued) 22. Powers DM, Boyd JC, Glick MR, et al. Interference Testing in Clinical
Chemistry; Proposed Guideline (EP7-P). Villanova, PA: The National
Method Comparison Committee for Clinical Laboratory Standards, 1986.
Correlation studies were performed using CLSI protocol NCCLS 23. Young DS. Effects of Drugs on Clinical Laboratory Tests, 4th ed.
EP9-A.25 Washington, DC: AACC Press; 1995:3-6–3-16.
Serum results from the ALT assay on the AEROSET System were 24. Kennedy JW, Carey RN, Coolen RB, et al. Evaluation of Precision
compared with those from a commercially available NADH oxidation Performance of Clinical Chemistry Devices; Approved Guideline
methodology. (EP5-A). Wayne, PA: The National Committee for Clinical Laboratory
Serum results from the ALT assay on an ARCHITECT c System were Standards, 1999.
compared with the ALT assay on the AEROSET System. 25. Kennedy JW, Carey RN, Coolen RB, et al. Method Comparison and
Bias Estimation Using Patient Samples; Approved Guideline (EP9-A).
AEROSET vs. ARCHITECT Wayne, PA: The National Committee for Clinical Laboratory
Comparative Method vs. AEROSET Standards, 1995.
N 74 83
TRADEMARKS
Y - Intercept -4.356 -3.683 The ARCHITECT c System family of instruments consists of c 4000,
Correlation Coefficient 0.989 1.000 c 8000, and c 16000 instruments.
Slope 0.870 0.940 AEROSET, ARCHITECT, c 4000, c 8000, c 16000, c System, and
SmartWash are trademarks of Abbott Laboratories in various
Range (U/L)* 4.8 to 130.0 7.0 to 3,935.2 jurisdictions.
*AEROSET Range All trademarks are property of their respective owner(s).

BIBLIOGRAPHY
1. US Department of Labor, Occupational Safety and Health
Administration. 29 CFR Part 1910.1030. Bloodborne Pathogens.
2. US Department of Health and Human Services. Biosafety in
Microbiological and Biomedical Laboratories, 5th ed. Washington,
DC: US Government Printing Office, December 2009.
3. World Health Organization. Laboratory Biosafety Manual, 3rd ed.
Geneva: World Health Organization, 2004.
4. Sewell DL, Bove KE, Callihan DR, et al. Protection of Laboratory
Workers from Occupationally Acquired Infections; Approved
Guideline—Third Edition (M29-A3). Wayne, PA: Clinical and
Laboratory Standards Institute, 2005.
5. Henry RJ, Cannon DC, Winkelman JW. Clinical Chemistry Principles
and Technics, 2nd ed. Hagerstown, MD: Harper and Row; 1974:888.
6. Burtis CA, Ashwood ER, editors. Tietz Textbook of Clinical Chemistry,
2nd ed. Philadelphia, PA: WB Saunders; 1994:795–7.
7. Williams K, Williams A, Kline L, et al. Stability of serum alanine
aminotransferase activity. Transfusion 1987;27(5):431–3.
8. Cuccherini B, Nussbaum S, Seeff L, et al. Stability of aspartate
aminotransferase and alanine aminotransferase activities.
J Lab Clin Med 1983;102(3):370–6.
9. Ruby SG, Relber NE, Lonser RE. Preanalytical variation in alanine
aminotransferase. Clin Chem 1988;34(4):744–5.
10. Heins M, Heil W, Withold W. Storage of serum and whole blood
samples? Effects of time and temperature on 22 serum analytes.
Eur J Clin Chem Clin Biochem 1995;33:231–8.
11. Dale JC, Pruett SK. Phlebotomy—a minimalist approach. Mayo Clin
Proc 1993;68(3):249–55.
12. Young D. Effects of Preanalytical Variables on Clinical Laboratory
Tests, 2nd ed. Washington, DC: AACC Press; 1997:3–12.
13. Elfath D, Cooney J, McDaniel R, et al. Effect of frozen storage of
serum on the level of 22 chemistry analytes. Clin Chem 1991;37:931.
14. Faulkner AM, Lukes-Hall AM, White GW. Evaluation of the Grenier
plasma separator blood tube. Ann Clin Biochem 1990;27:386–7.
15. Wilding P, Zilva JA, Wilde CE. Transport of specimens for clinical
chemistry analysis. Ann Clin Biochem 1977;14:301–6.
16. Schwartz MK. Interferences in diagnostic biochemical procedures.
Adv Clin Chem 1973;16:10.
17. Mosley JW, Goodwin RF. Stability of serum glutamic pyruvic
transaminase activity on storage. Am J Clin Pathol 1985;44:591–5.
18. Donnelly JG, Soldin SJ, Nealon DA, et al. Stability of twenty-five
analytes in human serum at 22°C, 4°C, and -20°C. Pediatr Pathol Lab
Med 1995;15:869–74.
19. Kaplan LA, Pesce AJ, editors. Clinical Chemistry Theory, Analysis,
and Correlation, 2nd ed. St Louis, MO: CV Mosby; 1989:895–8.
20. Sherman KE, Dodd RY, et al. Alanine aminotransferase levels among
volunteer blood donors: geographic variation and risk factors.
J Infect Dis 1982;145(3):383–6.
21. Passey RB, Bee DE, Caffo A, et al. Evaluation of the Linearity
of Quantitative Analytical Methods; Proposed Guideline (EP6-P). Abbott Laboratories ABBOTT
Villanova, PA: The National Committee for Clinical Laboratory Abbott Park, IL 60064 USA Max-Planck-Ring 2
Standards, 1986. 65205 Wiesbaden
September 2012 Germany
304663/R02 ©2009, 2012 Abbott Laboratories +49-6122-580

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ARCHITECT c SYSTEMS ASSAY PARAMETERS

Alanine Aminotransferase Serum/Plasma—Conventional and SI Units

Configure assay parameters — General Configure assay parameters — SmartWash

● General о Calibration о SmartWash о Results о Interpretation о General о Calibration ● SmartWash о Results о Interpretation
Assay: ALT Type: Photometric Version: † Assay: ALT
Number: 1021 COMPONENT REAGENT / ASSAY WASH Volume Replicates
Run controls for onboard reagents by:†† Lot Cuvette Trig* 10% Detergent B 345
● Reaction definition о Reagent / Sample о Validity checks
Reaction mode: Rate down *Not required for ARCHITECT Software version 7.00 and above.
Primary Secondary Read times
Wavelength: 340 / 380 Main: 21 – 33 Configure assay parameters — Results
Last required read: 33 Flex: 18 – 22
Absorbance range: 0.5000 – 1.5000 Color correction: 14 – 16 о General о Calibration о SmartWash ● Results о Interpretation
Sample blank type: None Assay: ALT Assay number: 1021
Dilution default range: Result units: U/L
Low-Linearity: 6 ‡‡
о Reaction definition ● Reagent / Sample о Validity checks High-Linearity: 942
R1 R2 Gender and age specific ranges:
Reagent: ALT00 Reagent volume: 160 40 GENDER AGE (UNITS) NORMAL EXTREME
Diluent: Saline Water volume: ___ ___ Either 0 – 130 (Y) 0 – 55
Diluent dispense mode: Type 0 Dispense mode: Type 0 Type 0
Diluted Default Configure result units
Dilution name Sample sample Diluent Water Dilution factor dilution
STANDARD : 5.3 ___ ___ ___ = 1:1.00 ● Assay: ALT
1:5 : 20.0 5.3 80 ___ = 1:5.00 о Version: †
_________ : ___ ___ ___ ___ = о Result units: U/L
Decimal places: 0 [Range 0 – 4]
Correlation factor: 1.0000
о Reaction definition о Reagent / Sample ● Validity checks Intercept: 0.0000
Reaction check: End Subtraction
A B
Read time: 1–1 2–2
Calculation limits: 0.0001 – 9.9999
Rate linearity %: 10

Configure assay parameters — Calibration

о General ● Calibration о SmartWash о Results о Interpretation
Assay: ALT Calibration method: Factor
Factor: ‡
● Calibrators о Volumes о Intervals о Validity checks
Calibrator set: Calibrator level: Concentration:
None Blank: Water 0

Replicates: 3 [Range 1 – 3]

о Calibrators ● Volumes о Intervals о Validity checks

Calibrator: Diluted
Calibrator level Sample sample Diluent Water
Blank: Water 5.3 ___ ___ ___

о Calibrators о Volumes ● Intervals о Validity checks

Calibration intervals:
Full interval: 648 (hours)

о Calibrators о Volumes о Intervals ● Validity checks

Blank absorbance range: _____ – _____

† Due to differences in instrument systems and unit configurations, version numbers may vary.
†† Parameter is available in ARCHITECT Software version 7.00 and above.
‡ The calibration factor for c 4000 and c 8000 is 8141; the calibration factor for c 16000 is 8492.
‡‡ The linear low value (Low-Linearity) is LOQ rounded up to the number of decimal places defined in the decimal places parameter field.

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