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GOT (ASAT) IFCC mod. liquiUV Calculation

U/l = (A2 – A0)/2 * Factor
Aspartate Aminotransferase (EC 2.6.1.1) Wavelength Temperature Factor for Sample Factor for Substrate
[nm] Start Procedure Start Procedure
Package Sizes
334 37°C 1780 2184
[REF] 12211 16 x 5 ml Reagent M- kit
12011 10 x 10 ml Reagent kit 340 37°C 1745 2143
12021 8 x 50 ml Reagent kit 365 37°C 3235 3971
12031 4 x 250 ml Reagent kit
If control results are outside the allowable ranges, the calculation factor should be
[IVD]
checked with a suitable calibrator material and adjusted using correction factors.
Method 1
Conversion from traditional units in SI-units:
Kinetic method for the determination of GOT (ASAT) activity according to the
1 U/l = 0.0167 μkat/l
recommendations of the Expert Panel of the IFCC (International Federation of Clinical
1 μkat/l = 60 U/l
Chemistry). Without pyridoxal phosphate activation.
Performance Characteristics
Reaction Principle
Measuring range up to 600 U/l or 10.0 μkat/l
GOT
2-Oxoglutarate + L-aspartate L-glutamate + oxaloacetate Samples with concentrations above the measuring range should be diluted (100 μl
sample + 900 μl physiological saline solution) and re-run. Multiply the result by 10.
MDH Samples with very high concentrations may produce falsely low results. In this case
Oxaloacetate + NADH + H+ L-malate + NAD+ dilute as described above.
Contents Interfering substances were added to a known sample. No interference was
[²REF²] 12211 12011 12021 12031 detected up to following concentrations:
[BUF] 16 x 4 ml 10 x 8 ml 8 x 40 ml 4 x 200 ml Ascorbic acid up to 20 mg/dl
[SUB] 1 x 16 ml 2 x 10 ml 8 x 10 ml 4 x 50 ml Bilirubin up to 40 mg/dl
[BUF] Buffer / Enzyme reagent Hemolysis (hemoglobin) up to 50 mg/dl; strong interference
TRIS buffer (pH 7.9) 100 mmol/l
Lipemia (Intralipid) up to 300 mg/dl
L-aspartate 300 mmol/l
LDH ≥ 1.13 kU/l Pyruvate up to 1 mmol/l
MDH ≥ 0.75 kU/l Typical performance data can be found in the Verification Report, accessible via:
Sodium azide 0.095 % www.human.de/data/gb/vr/en-gotli.pdf
[SUB] Substrate www.human-de.com/data/gb/vr/en-gotli.pdf
2-oxoglutarate 60 mmol/l If the performance data are not accessible via internet, they can be obtained free of
NADH 0.9 mmol/l charge from your local distributor.
Sodium azide 0.095 %
Reference Values 5,6
Reagent Preparation and Stability
Temperature 37°C IFCC*
Substrate Start Procedure
Men up to up to 37 U/l up to 35 U/l
The reagents are ready for use and stable, even after opening, up to the stated expiry
date when stored protected from light at 2 - 8°C. Contamination must be avoided! Women up to 31 U/l up to 31 U/l
Sample Start Procedure
*with pyridoxalphosphate activation
[²REF²] 12031 and 12021: Pour the entire contents of one bottle [SUB] into one bottle
Quality Control
[BUF], mix thoroughly.
All control sera with GOT values determined by this method can be employed.
[²REF²] 12211: Pipette 1 ml from bottle [SUB] into one bottle [BUF], mix thoroughly.
We recommend to use HumaTrol controls based on animal serum or SERODOS
[²REF²] 12011: Pipette 2 ml from bottle [SUB] into one bottle [BUF], mix thoroughly.
controls based on human serum.
The working reagent is stable for 4 weeks at 2 - 8°C or 5 days at 15 - 25°C.
Automation
Specimen
Proposals to apply the reagents on analysers are available on request. Each laboratory
Serum, heparinised plasma or EDTA plasma
has to validate the application in its own responsibility.
Loss of activity within 3 days at 4°C at 20 – 25°C
Precautionary statements
~ 8% ~ 10% P234 Keep only in original container.
Assay P260 Do not breathe dust/fume/gas/mist/vapours/spray.
Wavelength: 334, 340, or 365 nm P262 Do not get in eyes, on skin, or on clothing.
Optical path: 1 cm P281 Use personal protective equipment as required.
Temperature: 37°C P303+P361+P353 IF ON SKIN (or hair): Take off immediately all contaminated
Measurement: against air (decreasing absorbance) clothing. Rinse skin with water/shower.
P305+P351+P338 IF IN EYES: Rinse cautiously with water for several minutes. Remove
Warm the reagents and the cuvettes to 37°C. Temperature must be kept constant (±
contact lenses, if present and easy to do. Continue rinsing.
0.5°C) for the duration of the test.
P337+P313 If eye irritation persists: Get medical advice/attention.
Substrate Start Procedure* P401 Store in accordance with local/regional/national/international regulations.
Pipette into cuvettes 37°C P501 Dispose of contents/container in accordance with local/regional/national/
Sample 100 μl international regulations.

[BUF] 1000 μl References

1. Clin. Chim. Acta 70, 19-42 (1976)
Mix, incubate for 5 min at 37°C
2. Wallnöfer H. et al., Synopsis der Leberkrankheiten, Georg Thieme Verlag,
[SUB] 250 μl Stuttgart 1974
Mix, read the absorbance after 1 minute (A0) and at the same time start the stop 3. Thefeld, W. et al.; Dtsch. med. Wschr. 99, 343 (1974)
watch. Read the absorbance again exactly after 2 minutes (A2). 4. Schumann, G. et al., Clin. Chem. Lab. Med. 40, 725-733 (2002)
* Semi micro method; for macro methods double the volumes. 5. Schumann, G., Klauke, R., Clin. Chim. Acta 327, 69-79 (2003)
Sample Start Procedure* 6. Fischbach, F., Zawta, B., Klin. Lab. 38, 555-561 (1992)
Pipette into cuvettes 37°C 7. Thomas L., Labor und Diagnose, 8. ed., TH-Books, 78-98 (2012)
8. IFCC AST Reference procedure, CCLM, part 5, 2002; 40 (7): 725-733
Sample 100 μl
9. Young D.S., Effects of Drugs on Clinical Laboratory Tests, 5. ed, AACC Press
Working reagent 1000 μl (2000)
10. DGKL, Die Qualität diagnostischer Proben, 7. ed, Heidelberg (2012)
Mix, read the absorbance after 1 minute (Ao) and at the same time start the stop
watch. Read the absorbance again exactly after 2 minutes (A2).
* Semi micro method; for macro methods double the volumes.
EN-GOTLI INF 1221101 GB 08-2021-023 |
The pipetting scheme for HumaLyzer 4000 can be accessed via:
www.human.de/aps-cc
Human Gesellschaft für Biochemica und Diagnostica mbH
Max-Planck-Ring 21 · 65205 Wiesbaden · Germany
Telefon +49 6122-9988-0 · Telefax +49 6122-9988-100 · e-Mail human@human.de