ACTIVATED ALANINE AMINOTRANSFERASE

en
8L92-21 and 8L92-41
307229/R06
B8L9X0

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

Read Highlighted Changes: Revised March 2017.

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

Batch code/Lot number Manufacturer

Manufactured for Sufficient for

Product of Japan Temperature limitation

Reagent 1 Use by/Expiration date

1
NAME REAGENT HANDLING AND STORAGE
ACTIVATED ALANINE AMINOTRANSFERASE Reagent Handling
Remove air bubbles, if present in the reagent cartridge, with a new
INTENDED USE applicator stick. Alternatively, allow the reagent to sit at the appropriate
The Activated Alanine Aminotransferase (Activated ALT) assay is used storage temperature to allow the bubbles to dissipate. To minimize
for the quantitation of alanine aminotransferase in human serum or volume depletion, do not use a transfer pipette to remove the bubbles.
plasma.
CAUTION: Reagent bubbles may interfere with proper detection of
reagent level in the cartridge, causing insufficient reagent aspiration
SUMMARY AND EXPLANATION OF TEST which could impact results.
Alanine Aminotransferase (ALT), also referred to as glutamate pyruvate
transaminase (GPT), is an enzyme involved in amino acid metabolism. Reagent Storage
It is found in many tissues, but the highest levels are found in the liver Unopened reagents are stable until the expiration date when stored
and kidney. Tissue destruction leads to the release of the intracellular at 2 to 8°C.
enzyme into the circulating blood. Markedly elevated serum ALT levels Reagent stability is 30 days if the reagent is uncapped and onboard.
may be found in a variety of diseases of the liver, such as hepatitis,
mononucleosis, and cirrhosis. These very high levels of ALT are not Indications of Deterioration
usually observed in other disease processes, e.g., myocardial infarction; Instability or deterioration should be suspected if there are precipitates,
thus, ALT is regarded as a reasonably specific indicator of liver visible signs of leakage, extreme turbidity, microbial growth, if calibration
disease. does not meet the appropriate package insert and/or ARCHITECT
System Operations Manual criteria, or if controls do not meet the
PRINCIPLES OF PROCEDURE appropriate criteria.
ALT present in the sample catalyzes the transfer of the amino
group from L-alanine to 2-oxoglutarate, in the presence of WARNINGS AND PRECAUTIONS
pyridoxal‑5'‑phosphate, forming pyruvate and L-glutamate. Pyruvate Precautions for Users
in the presence of NADH and lactate dehydrogenase (LD) is reduced
to L-lactate. In this reaction NADH is oxidized to NAD. The reaction is •
monitored by measuring the rate of decrease in absorbance at 340 nm • For In Vitro Diagnostic Use.
due to the oxidation of NADH to NAD. • Do not use components beyond the expiration date.
ALT, P-5'-P • Do not mix reagents from different kit lot numbers.
L-Alanine + 2-Oxoglutarate Pyruvate + L-Glutamate • CAUTION: This product requires the handling of human specimens.
LD It is recommended that all human-sourced materials be considered
Pyruvate + NADH L-Lactate + NAD+ potentially infectious and handled in accordance with the OSHA
Standard on Bloodborne Pathogens.3 Biosafety Level 24 or other
The Activated ALT reagent is based on the optimized formulation as
appropriate biosafety practices5,6 should be used for materials
recommended by the International Federation of Clinical Chemistry
that contain or are suspected of containing infectious agents.
(IFCC).1,2
• Safety Data Sheets are available at www.abbottdiagnostics.com or
Methodology: IFCC contact your local representative.
REAGENTS • For a detailed discussion of safety precautions during system
operation, refer to the ARCHITECT System Operations Manual,
Reagent Kit Section 8.
Activated ALT is supplied as a liquid, ready-to-use, two-reagent kit which
contains: SPECIMEN COLLECTION AND HANDLING
8L92-21 Suitable Specimens
10 x 40 mL Serum and plasma are acceptable specimens.
10 x 40 mL CAUTION: Erythrocytes contain approximately seven times more ALT
Estimated tests per kit: 2,750* than does serum.7 Hemolysis in serum or plasma can increase test
results.
8L92-41 • Serum: Use serum collected by standard venipuncture techniques
10 x 68 mL into glass or plastic tubes with or without gel barriers. Ensure
complete clot formation has taken place prior to centrifugation.
10 x 68 mL Centrifuge according to tube manufacturer’s instructions to ensure
Estimated tests per kit: 4,820* proper separation of serum from blood cells.
*Calculation is based on the minimum reagent fill volume per kit. Some specimens, especially those from patients receiving
anticoagulant or thrombolytic therapy, may take longer to complete
Reactive Ingredients Concentration their clotting processes. Fibrin clots may subsequently form in these
β-NADH 0.369 mmol/L sera and the clots could cause erroneous test results.
• Plasma: Use plasma collected by standard venipuncture techniques
LD 2,460 U/L
into glass or plastic tubes. Acceptable anticoagulants are lithium
Pyridoxal phosphate monohydrate 0.205 mmol/L heparin (with or without gel barrier), sodium heparin, and EDTA.
TRIS 100 mmol/L Ensure centrifugation is adequate to remove platelets. Centrifuge
according to tube manufacturer’s instructions to ensure proper
L-alanine 1,025 mmol/L separation of plasma from blood cells.
2-oxoglutaric acid 30.75 mmol/L For total sample volume requirements, refer to the ASSAY PARAMETERS
section of this package insert and Section 5 of the ARCHITECT System
TRIS 105 mmol/L Operations Manual.

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Specimen Storage
Serum and Plasma
It is recommended that specimens be assayed on the day of
collection.8,9 Numerous publications have defined storage conditions
for ALT.10-21 Examples are shown in the table below.
Temperature Maximum Bibliographic
Storage Reference
30°C 3 days 15
2 to 8°C 7 days 15
-40°C 60 days 20
When samples were stored at -20°C for 8 days, an 11% reduction in ALT
activity was observed; a 20% reduction in ALT activity was observed
when specimens were stored at -20°C for 1 month.21
NOTE: Stored specimens must be inspected for particulates. If present,
mix and centrifuge the specimen to remove particulates prior to testing.
PROCEDURE
Materials Provided
8L92 Activated ALT Reagent Kit
Materials Required but not Provided
• Control Material
• Saline (0.85% to 0.90% NaCl) for specimens that require dilution
Assay Procedure
For a detailed description of how to run an assay, refer to Section 5 of
the ARCHITECT System Operations Manual.
Specimen Dilution Procedures
The ARCHITECT c Systems have an automatic dilution feature; refer to
Section 2 of the ARCHITECT System Operations Manual for additional
information.
Serum and Plasma: Specimens with ALT values exceeding 2,855 U/L
(4,772 U/L for Flex Rate Linearity) are flagged and may be diluted by
following either the Automated Dilution Protocol or the Manual Dilution
Procedure.
Automated Dilution Protocol
If using the Automated Dilution Protocol, the system performs a 1:5
dilution of the specimen and automatically corrects the enzyme activity
value by multiplying the result by the appropriate dilution factor.
Manual Dilution Procedure
Manual dilutions should be performed as follows:
• Use saline (0.85% to 0.90% NaCl) to dilute the sample.
• The operator must enter the dilution factor in the patient or control
order screen. The system uses this dilution factor to automatically
correct the enzyme activity value by multiplying the result by the
entered factor.
• If the operator does not enter the dilution factor, the result must
be multiplied by the appropriate dilution factor before reporting the
result.
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
dilution.
For detailed information on ordering dilutions, refer to Section 5 of the
ARCHITECT System Operations Manual.
CALIBRATION
Calibration is stable for approximately 30 days (720 hours) and is
required with each change in reagent lot number. Verify calibration
with at least two levels of controls according to the established quality
control requirements for your laboratory. If control results fall outside
acceptable ranges, recalibration may be necessary.
A calibration factor was established based on the IFCC methodology.2 A
calibration factor must be entered on the Configure assay parameters
window, Calibration view. Refer to the ASSAY PARAMETERS section of
this package insert for the specific factor.
For a detailed description of how to calibrate an assay, refer to Section 6
of the ARCHITECT System Operations Manual.
3
QUALITY CONTROL
The following is the recommendation of Abbott Laboratories for quality
control. As appropriate, refer to your laboratory standard operating
procedure(s) and/or quality assurance plan for additional quality control
requirements and potential corrective actions.
• Two levels of controls (normal and abnormal) are to be run every
24 hours.
• If more frequent control monitoring is required, follow the established
quality control procedures for your laboratory.
• If quality control results do not meet the acceptance criteria
defined by your laboratory, patient values may be suspect. Follow
the established quality control procedures for your laboratory.
Recalibration may be necessary.
• Review quality control results and acceptance criteria following a
change of reagent lot.
RESULTS
Refer to Appendix C of the ARCHITECT System Operations Manual for
information on results calculations.
Representative performance data are given in the EXPECTED VALUES
and SPECIFIC PERFORMANCE CHARACTERISTICS sections of this
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.
All Activated ALT results, including < 6 U/L, should be evaluated by
comparing to other clinically relevant information.
EXPECTED VALUES
Reference Range
Serum22/Plasma
Range (U/L)
Adult 0 to 55
It is recommended that each laboratory determine its own reference
range based upon its particular locale and population characteristics.
Published reference range studies have demonstrated that ALT activity
varies with body-mass index (BMI), demographics, and geography.23,24
Using a population rigorously selected to include only healthy subjects,
investigators found upper reference limits of 19 U/L for females and
30 U/L for males.25 Preliminary upper reference limits based on a
hospitalized German population have been published for the IFCC
reference procedure. For ALT, these values were 34 U/L for females and
45 U/L for males.26
SPECIFIC PERFORMANCE CHARACTERISTICS
Linearity
Activated ALT is linear up to 2,855 U/L.
Flex Rate Linearity is 4,772 U/L. To use Flex Rate Linearity, the operator
must edit the linear high value to 4,772 on the Configure assay
parameters screen, Results view in the ARCHITECT c System ASSAY
PARAMETERS.
Linearity was verified using Clinical and Laboratory Standards Institute
(CLSI) protocol NCCLS EP6‑A.27
Limit of Detection (LOD)
The LOD for Activated ALT is 5 U/L. The LOD is the mean concentration
of an analyte-free sample + 2 SD, where SD = the pooled, within-run
standard deviation of the analyte-free sample. A study performed on an
ARCHITECT c System produced an LOD for Activated ALT of 4.8 U/L.
Limit of Quantitation (LOQ)
The LOQ for Activated ALT is 5.7 U/L. The LOQ is the analyte
concentration at which the CV = 20%.
Interfering Substances BIBLIOGRAPHY
Interference effects were assessed by Dose Response and Paired 1. Bergmeyer HU, Hørder M, Rej R. Approved recommendation (1985)
Difference methods, at the medical decision level of the analyte. on IFCC methods for the measurement of catalytic concentration of
enzymes. J Clin Chem Clin Biochem 1986;24(7):481–95.
Interfering Interferent Concentration N Target Observed 2. Schumann G, Bonora R, Ceriotti F, et al. IFCC primary reference
procedures for the measurement of catalytic activity concentrations
Substance (U/L) (% of Target) of enzymes at 37 degrees C. International Federation of Clinical
Chemistry and Laboratory Medicine. Part 4. Reference procedure for the
30 mg/dL (513 µmol/L) 4 58.5 101 measurement of catalytic concentration of alanine aminotransferase. Clin
Bilirubin Chem Lab Med 2002;40(7):718–24.
60 mg/dL (1,026 µmol/L) 4 58.5 105
3. US Department of Labor, Occupational Safety and Health Administration.
750 mg/dL (7.5 g/L) 4 63.8 105 29 CFR Part 1910.1030. Bloodborne Pathogens.
Hemoglobin 4. US Department of Health and Human Services. Biosafety in
1000 mg/dL (10.0 g/L) 4 63.8 106 Microbiological and Biomedical Laboratories, 5th ed. Washington, DC: US
Government Printing Office, December 2009.
500 mg/dL (5.0 g/L) 4 63.9 98 5. World Health Organization. Laboratory Biosafety Manual, 3rd ed. Geneva:
Intralipid World Health Organization, 2004.
750 mg/dL (7.5 g/L) 3 63.9 99
6. Clinical and Laboratory Standards Institute (CLSI). Protection of
Bilirubin solutions at the above concentrations were prepared by addition Laboratory Workers From Occupationally Acquired Infections; Approved
of a bilirubin stock to human serum pools. Hemoglobin solutions at Guideline—Fourth Edition. CLSI Document M29-A4. Wayne, PA: CLSI;
2014.
the above concentrations were prepared by addition of hemolysate to 7. Rej R. Aminotransferases in disease. Clin Lab Med 1989;9(4):667–87.
human serum pools. Intralipid solutions at the above concentrations were 8. Burtis CA, Ashwood ER, Bruns DE, editors. Tietz Textbook of Clinical
prepared by addition of Intralipid to human serum pools. Chemistry and Molecular Diagnostics, 4th ed. St. Louis, MO: Elsevier;
The following drugs were tested for interference at the concentrations 2006:604–7.
indicated using an acceptance criteria of ± 10% from the target value. 9. Cuccherini B, Nussbaum SJ, Seeff LB, et al. Stability of aspartate
aminotransferase and alanine aminotransferase activities. J Lab Clin Med
Interfering Interferent Target Observed 1983;102(3):370–6.
N 10. Williams KM, Williams AE, Kline LM, et al. Stability of serum alanine
Substance Concentration (U/L) (% of Target) aminotransferase activity. Transfusion 1987;27(5):431–3.
Sulfapyridine 60 mg/L (241.0 µmol/L) 3 50.3 94 11. Henry RJ, Cannon DC, Winkelman JW. Clinical Chemistry Principles and
Technics, 2nd ed. Hagerstown, MD: Harper and Row; 1974:888.
Sulfasalazine 20 mg/L (50.3 µmol/L) 3 50.3 91 12. Ruby SG, Reiber NE, Lonser RE. Pre-analytical variation in alanine
aminotransferase. Clin Chem 1988;34(4)744–5.
Temozolomide 20 mg/L (103.1 µmol/L) 3 78.6 103 13. Heins M, Heil W, Withold W. Storage of serum or whole blood samples?
Effects of time and temperature on 22 serum analytes. Eur J Clin Chem
Interferences from medications or endogenous substances may affect Clin Biochem 1995;33(4):231–8.
results.28 14. Dale JC, Pruett SK. Phlebotomy—a minimalist approach. Mayo Clin Proc
1993;68(3):249–55.
Precision 15. Young DS. Effects of Preanalytical Variables on Clinical Laboratory Tests,
The imprecision of the Activated ALT assay is ≤ 6.0% Total CV. 2nd ed. Washington, DC: AACC Press; 1997:3-12.
Representative data from studies using CLSI protocol NCCLS EP5-A229 16. Elfath D, Cooney J, McDaniel R, et al. Effect of frozen storage of serum
are summarized below. on the level of 22 chemistry analytes [Poster 0102 presented at AACC
43rd National Meeting in Washington, DC: July 30, 1991]. Clin Chem
1991;37(6):931.
Control Level 1 Level 2 17. Faulkner AM, Lukes-Hall AM, White GW. Evaluation of the Greiner
N 80 80 plasma separator blood tube. Ann Clin Biochem 1990;27:386–7.
18. Wilding P, Zilva JF, Wilde CE. Transport of specimens for clinical
Mean (U/L) 37 85 chemistry analysis. Ann Clin Biochem 1977;14(6):301–6.
19. Schwartz MK. Interferences in diagnostic biochemical procedures.
SD 0.45 0.42 Adv Clin Chem 1973;16:10.
Within Run 20. Mosley JW, Goodwin RF. Stability of serum glutamic pyruvic
%CV 1.2 0.5 transaminase activity on storage. Am J Clin Pathol 1965;44:591–5.
SD 0.44 0.26 21. Donnelly JG, Soldin SJ, Nealon DA, et al. Stability of twenty-five analytes
Between Run in human serum at 22°C, 4°C, and -20°C. Pediatr Pathol Lab Med
%CV 1.2 0.3 1995;15(6):869–74.
22. Murray RL. Alanine aminotransferase. In: Kaplan LA, Pesce AJ,
SD 0.65 1.41 editors. Clinical Chemistry Theory, Analysis, and Correlation, 2nd ed.
Between Day St. Louis, MO: CV Mosby; 1989:895–8.
%CV 1.8 1.7 23. Sherman KE, Dodd RY, et al. Alanine aminotransferase levels among
volunteer blood donors: geographic variation and risk factors.
SD 0.90 1.50 J Infect Dis 1982;145(3):383–6.
Total 24. Sherman KE. Alanine aminotransferase in clinical practice. A review.
%CV 2.5 1.8 Arch Intern Med 1991;151(2);260–5.
25. Prati D, Taioli E, Zanella A, et al. Updated definitions of healthy ranges
Method Comparison for serum alanine aminotransferase levels. Ann Intern Med 2002;
Correlation studies were performed using CLSI protocol NCCLS 137(1):1–10.
26. Schumann G, Klauke R. New IFCC reference procedures for the
EP9‑A2.30 determination of catalytic activity concentrations of five enzymes in
Serum results from the Activated ALT assay on an ARCHITECT c System serum: preliminary upper reference limits obtained in hospitalized
were compared with those from the ALT Activated (ALT-A 8D36) assay subjects. Clin Chim Acta 2003;327(1–2):69–79.
on the AEROSET System. 27. Tholen DW, Kroll M, Astles JR, et al. Evaluation of the Linearity of
Quantitative Measurement Procedures: A Statistical Approach; Approved
Guideline (EP6-A). Wayne, PA: The National Committee for Clinical
Activated ALT on ARCHITECT Laboratory Standards, 2003.
vs. 28. Young DS. Effects of Drugs on Clinical Laboratory Tests, 4th ed.
Abbott ALT-A on AEROSET Washington, DC: AACC Press; 1995:3-6–3-16.
29. Tholen DW, Kallner A, Kennedy JW, et al. Evaluation of Precision
N 106 Performance of Quantitative Measurement Methods; Approved
Guideline—Second Edition (EP5-A2). Wayne, PA: The National
Y - Intercept 7.72 Committee for Clinical Laboratory Standards, 2004.
Correlation Coefficient 0.998 30. Krouwer JS, Tholen DW, Garber CC, et al. Method Comparison and
Bias Estimation Using Patient Samples; Approved Guideline—Second
Slope 1.00 Edition (EP9-A2). Wayne, PA: The National Committee for Clinical
Range (U/L) 10 to 4,564 Laboratory Standards, 2002.

Abbott Laboratories
Diagnostics Division
Abbott Park, IL 60064 USA
March 2017
©2009, 2017 Abbott Laboratories
TRADEMARKS
The ARCHITECT c System family of instruments consists of c 4000,
Abbott GmbH & Co. KG c 8000, and c 16000 instruments.
Max-Planck-Ring 2 AEROSET, ARCHITECT, c 4000, c 8000, c 16000, c System, and
65205 Wiesbaden SmartWash are trademarks of Abbott Laboratories in various
Germany
+49-6122-580 jurisdictions.
All other trademarks are property of their respective owners.

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ARCHITECT c Systems ASSAY PARAMETERS

Activated 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: A-ALT Type: Photometric Version: † Assay: A-ALT
Number: 1032 COMPONENT REAGENT / ASSAY WASH Volume Replicates
Run controls for onboard reagents by: Lot
● Reaction definition о Reagent / Sample о Validity checks
Reaction mode: Rate down
Primary Secondary Read times Configure assay parameters — Results
Wavelength: 340 / 548 Main: 21 – 33
Last required read: 33 Flex: 18 – 21 о General о Calibration о SmartWash ● Results о Interpretation
Absorbance range: 0.8000 – 3.0000 Color correction: __ – __ Assay: A-ALT Assay number 1032
Sample blank type: None Dilution default range: Result units: U/L
Low-Linearity: 6**
High-Linearity: 2855
о Reaction definition ● Reagent / Sample о Validity checks Gender and age specific ranges:*
R1 R2 GENDER AGE (UNITS) NORMAL EXTREME
Reagent: AALT9 Reagent volume: 120 120 Either 0 – 130 (Y) 0 – 55
Diluent: Saline Water volume: ___
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 Assay: A-ALT
STANDARD : 6.0 ___ ___ ___ = 1:1.00 ● Version: †
1:5 : 20.0 6.0 80 ___ = 1:5.00 о Result units: U/L
_________ : ___ ___ ___ ___ = о Decimal places: 0 [Range 0 – 4]
Correlation factor: 1.0000
о Reaction definition о Reagent / Sample ● Validity checks Intercept: -10.0000
Reaction check: None

Rate linearity %: 10

Configure assay parameters — Calibration

о General ● Calibration о SmartWash о Results о Interpretation
Assay: A-ALT Calibration method: Factor
Factor: 7658.0000
● 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 6.0 ___ ___ ___

о Calibrators о Volumes ● Intervals о Validity checks

Calibration intervals:
Full interval: 720 (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.
** The linear low value (Low-Linearity) is LOQ rounded up to the number of decimal places defined in the decimal places parameter field.
* User defined.

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