20764957322V2

ALTL

Alanine Aminotransferase acc. to IFCC without pyridoxal phosphate activation

Indicates cobas c systems on which reagents can be used
Order information
Alanine Aminotransferase acc. to IFCC
500 tests
Calibrator f.a.s. (12 x 3 mL)
Calibrator f.a.s. (12 x 3 mL, for USA)
Precinorm U plus (10 x 3 mL)
Precipath U plus (10 x 3 mL)
Precinorm U (20 x 5 mL)
Precipath U (20 x 5 mL)
NaCl Diluent 9% (50 mL)

Cat. No. 20764957 322

Cat. No. 10759350 190
Cat. No. 10759350 360
Cat. No. 12149435 122
Cat. No. 12149443 122
Cat. No. 10171743 122
Cat. No. 10171778 122
Cat. No. 04489357 190

System-ID 07 6495 7
Code 401
Code 401
Code 300
Code 301
Code 300
Code 301
System-ID 07 6869 3

English

Storage and stability

System information
ALTL: ACN 685

ALTL
Shelf life at 2-8C:

Intended use
In vitro test for the quantitative determination of alanine aminotransferase
(ALT) in human serum and plasma on Roche/Hitachi cobas c systems.
Summary1,2

The enzyme alanine aminotransferase (ALT) has been widely reported as

present in a variety of tissues. The major source of ALT is the liver, which
has led to the measurement of ALT activity for the diagnosis of hepatic
diseases. Elevated serum ALT is found in hepatitis, cirrhosis, obstructive
jaundice, carcinoma of the liver, and chronic alcohol abuse. ALT is only slightly
elevated in patients who have an uncomplicated myocardial infarction.
Although both serum aspartate aminotransferase (AST) and ALT become
elevated whenever disease processes affect liver cell integrity, ALT is
the more liver-specific enzyme. Moreover, elevations of ALT activity
persist longer than elevations of AST activity.
In patients with vitamin B6 deficiency, serum aminotransferase activity may be
decreased. The apparent reduction in aminotransferase activity may be related
to decreased pyridoxal phosphate, the prosthetic group for aminotransferases,
resulting in an increase in the ratio of apoenzyme to holoenzyme.
Test principle
This assay follows the recommendations of the IFCC, but was
optimized for performance and stability.3,4,5
ALT catalyzes the reaction between L-alanine and 2-oxoglutarate. The
pyruvate formed is reduced by NADH in a reaction catalyzed by lactate
dehydrogenase (LDH) to form L-lactate and NAD+.
L-Alanine + 2-oxoglutarate

ALT

pyruvate + L-glutamate

Pyruvate + NADH + H+

LDH

L-lactate + NAD+

The rate of the NADH oxidation is directly proportional to the catalytic ALT
activity. It is determined by measuring the decrease in absorbance.
Reagents - working solutions
R1 TRIS buffer: 224 mmol/L, pH 7.6 (25C); L-alanine: 1120 mmol/L;
albumin (bovine): 0.25%; LDH (microorganisms): 45 kat/L; stabilizers;
preservative
R2 2-Oxoglutarate: 94 mmol/L; NADH: 1.7 mmol/L; additives; preservative

Roche/Hitachi cobas c systems

cobas c 501

See expiration date on

cobas c pack label.
On-board in use and refrigerated on the analyzer: 12 weeks
NaCl Diluent 9%
Shelf life at 2-8C:

See expiration date on

cobas c pack label.
On-board in use and refrigerated on the analyzer: 12 weeks

Specimen collection and preparation

For specimen collection and preparation, only use suitable
tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Serum (free from hemolysis).
Plasma (free from hemolysis): Li-heparin or K2-EDTA plasma.
The sample types listed were tested with a selection of sample collection tubes
that were commercially available at the time of testing, i.e. not all available
tubes of all manufacturers were tested. Sample collection systems from
various manufacturers may contain differing materials which could affect
the test results in some cases. When processing samples in primary tubes
(sample collection systems), follow the instructions of the tube manufacturer.
Separate the serum or plasma from the clot or cells promptly.
Centrifuge samples containing precipitates before performing the assay.
Stability: 3 days at 15-25C6,7
7 days at 2-8C6,7
>7 days at -70C7
Materials provided
See Reagents - working solutions section for reagents.
Materials required (but not provided)
See Order information section.
Distilled water
General laboratory equipment
Assay
For optimal performance of the assay, follow the directions given in this
document for the analyzer concerned. Refer to the appropriate operator
manual for analyzer-specific assay instructions.
The performance of applications not validated by Roche is not
warranted and must be defined by the user.

Precautions and warnings

For in vitro diagnostic use.
Exercise the normal precautions required for handling all laboratory reagents.
Safety data sheet available for professional user on request.
Disposal of all waste material should be in accordance with local guidelines.
Reagent handling
Ready for use.
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cobas c systems

ALTL

Alanine Aminotransferase acc. to IFCC without pyridoxal phosphate activation

Application for serum and plasma
cobas c 501 test definition
Assay type
Reaction time / Assay points
Wavelength (sub/main)
Reaction direction
Units

Rate A
10 / 18-46
700/340 nm
Decrease
U/L (kat/L)

Reagent pipetting
R1
R2

59 L
17 L

Sample volumes

Sample

Normal
Decreased
Increased

9 L
9 L
18 L

Special wash requirements

No interfering assays are known which require special wash steps.

Diluent (H2O)
32 L
20 L
Sample dilution
Sample
Diluent (NaCl)

15 L
135 L

Calibration
Calibrators
Calibration mode
Calibration frequency

S1: H2O
S2: C.f.a.s.
Linear
2-point calibration
- after reagent lot change
- and as required following quality control
procedures

Traceability: This method has been standardized against the original IFCC
formulation using calibrated pipettes together with a manual photometer
providing absolute values and the substrate-specific absorptivity, .3,5
Quality control
For quality control, use control materials as listed in the
Order information section.
Other suitable control material can be used in addition.
The control intervals and limits should be adapted to each laboratorys
individual requirements. Values obtained should fall within the defined limits.
Each laboratory should establish corrective measures to be taken
if values fall outside the limits.
Calculation
Roche/Hitachi cobas c systems automatically calculate the
analyte activity of each sample.
Conversion factor: U/L x 0.0167 = kat/L
Limitations - interference8
Criterion: Recovery within 10% of initial value at an ALT
activity of 30 U/L (0.5 kat/L).
Icterus: No significant interference up to an I index of 60 for conjugated
and unconjugated bilirubin (approximate conjugated and unconjugated
bilirubin concentration: 1026 mol/L (60 mg/dL)).
Hemolysis: No significant interference up to an H index of 200 (approximate
hemoglobin concentration: 124 mol/L (200 mg/dL)).
Contamination with erythrocytes will elevate results, because the analyte level
in erythrocytes is higher than in normal sera. The level of interference may be
variable depending on the content of analyte in the lysed erythrocytes.
Lipemia (Intralipid): No significant interference up to an L index
of 150. There is poor correlation between the L index (corresponds
to turbidity) and triglycerides concentration.
Lipemic samples may cause >Abs flagging. Choose diluted
sample treatment for automatic rerun.
Drugs: No interference was found using common drug panels.9
Exception: Calcium dobesilate causes artificially low ALT values
at the therapeutic drug level.
In very rare cases gammopathy, in particular type IgM (Waldenstrms
macroglobulinemia), may cause unreliable results.
For diagnostic purposes, the results should always be assessed in conjunction
with the patients medical history, clinical examination and other findings.
cobas c systems

Measuring range
5-700 U/L (0.08-11.7 kat/L)
Extended measuring range (calculated)
5-7000 U/L (0.08-117 kat/L)
Lower detection limit
5 U/L (0.08 kat/L)
The lower detection limit represents the lowest measurable analyte
level that can be distinguished from zero. It is calculated as the value
lying three standard deviations above that of the lowest standard
(standard 1 + 3 SD, within-run precision, n = 21).
Expected values10
Acc. to the optimized standard method (comparable to the IFCC
method without pyridoxal phosphate activation11):
Males
Females

up to 41 U/L up to 0.68 kat/L

up to 33 U/L up to 0.55 kat/L

Calculated values: a factor of 1.85 is used for the conversion

from 25C to 37C.12
Each laboratory should investigate the transferability of the expected values to
its own patient population and if necessary determine its own reference ranges.
Specific performance data
Representative performance data on the analyzers are given below.
Results obtained in individual laboratories may differ.
Precision
Reproducibility was determined using human samples and controls in
an internal protocol (within-run n = 21, total n = 60).
The following results were obtained:
Within-run
Precinorm U
Precipath U
Human serum 1
Human serum 2
Total
Precinorm U
Precipath U
Human serum 3
Human serum 4

Mean
U/L (kat/L)
39.5 (0.66)
120 (2.00)
113 (1.89)
7.2 (0.12)

SD
U/L (kat/L)
0.3 (0.01)
0.5 (0.01)
0.5 (0.01)
0.7 (0.01)

CV
%
0.6
0.4
0.4
9.3

Mean
U/L (kat/L)
39.3 (0.66)
120 (2.00)
24.0 (0.40)
98.1 (1.64)

SD
U/L (kat/L)
0.6 (0.01)
1.2 (0.02)
0.6 (0.01)
3.2 (0.05)

CV
%
1.4
1.0
2.6
3.3

Method comparison
ALT values for human serum and plasma samples obtained on a Roche/Hitachi
cobas c 501 analyzer (y) were compared with those determined using
the same reagent on a Roche/Hitachi 917 analyzer (x).
Sample size (n) = 198
Passing/Bablok13
y = 1.000x - 0.29 U/L
= 0.924

Linear regression
y = 0.997x - 1.05 U/L
r = 0.996

The sample activities were between 4.6 and 383 U/L (0.08 and 6.4 kat/L).
References
1. Sherwin JE. Liver function. In: Kaplan LA, Pesce AJ, eds.
Clinical Chemistry, theory, analysis, and correlation. St. Louis:
Mosby 1984:420-438.
2. Moss DW, Henderson AR, Kachmar JF. Enzymes. In: Tietz NW,
ed. Fundamentals of Clinical Chemistry. 3rd ed. Philadelphia:
WB Saunders 1987:346-421.
3. Bergmeyer HU, Hrder M, Rej R. Approved recommendation (1985)
on IFCC methods for the measurement of catalytic concentration
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ALTL

Alanine Aminotransferase acc. to IFCC without pyridoxal phosphate activation

4.
5.

6.
7.
8.
9.
10.

11.
12.
13.

of enzymes. Part 3. IFCC method for alanine aminotransferase.

J Clin Chem Clin Biochem 1986;24:481-495.
ECCLS. Determination of the catalytic activity concentration
in serum of L-alanine aminotransferase (EC 2.6.1.2, ALAT)
Klin Chem Mitt 1989;20:204-211.
Schumann G et al. IFCC Primary Reference Procedures for the
Measurement of Catalytic Activity Concentrations of Enzymes
at 37C Part 4. Reference Procedure for the Measurement of
Catalytic Activity Concentrations of Alanine Aminotransferase.
Clin Chem Lab Med 2002;40(7):718-724.
Heins M, Heil H, Withold W. Storage of Serum or Whole Blood
Samples? Effect of Time and Temperature on 22 Serum Analytes.
Eur J Clin Chem Clin Biochem 1995;33:231-238.
Data on file at Roche Diagnostics.
Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences
in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-474.
Report on the Symposium Drug effects in clinical chemistry methods,
Breuer J, Eur J Clin Chem Clin Biochem 1996;34:385-386.
Thefeld W, Hoffmeister H, Busch, E-W, Koller PU, Vollmar J.
Referenzwerte fr die Bestimmungen der Transaminasen GOT und
GPT sowie der alkalischen Phosphatase im Serum mit optimierten
Standardmethoden. Dtsch Med Wschr 1974;99:343-351.
Klein G, Lehmann P, Michel E, Regenauer H. Vergleich der
IFCC-Methoden fr ALAT, ASAT und GGT bei 37C mit den eingefhrten
Standardmethoden bei 25C und 37C. Lab Med 1994;18:403-404.
Zawta B, Klein G, Bablok W. Temperaturumrechnung in der
klinischen Enzymologie? Klin Lab 1994;40:23-32.
Bablok W et al. A General Regression Procedure for Method
Transformation. J Clin Chem Clin Biochem 1988;26:783-790.

FOR US CUSTOMERS ONLY: LIMITED WARRANTY

Roche Diagnostics warrants that this product will meet the specifications
stated in the labeling when used in accordance with such labeling and
will be free from defects in material and workmanship until the expiration
date printed on the label. THIS LIMITED WARRANTY IS IN LIEU OF ANY
OTHER WARRANTY, EXPRESS OR IMPLIED, INCLUDING ANY IMPLIED
WARRANTY OF MERCHANTABILITY OR FITNESS FOR PARTICULAR
PURPOSE. IN NO EVENT SHALL ROCHE DIAGNOSTICS BE LIABLE FOR
INCIDENTAL, INDIRECT, SPECIAL OR CONSEQUENTIAL DAMAGES.

COBAS, COBAS C, PRECINORM and PRECIPATH are trademarks of Roche.

Other brand or product names are trademarks of their respective holders.
Significant additions or changes are indicated by a change bar in the margin.
2006 Roche Diagnostics

Roche Diagnostics GmbH, D-68298 Mannheim

for USA: US Distributor:
Roche Diagnostics, Indianapolis, IN
US Customer Technical Support 1-800-428-2336

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