Biosystems Alaninaminotransferaz (ALT/GPT)
PRINCIPLE OF THE METHOD
- Alanine aminotransferase (ALT or GPT) catalyzes the transfer of the amino group from alanine
to 2-oxoglutarate, forming pyruvate and glutamate.
- The catalytic concentration is determined from the rate of decrease of NADH, me
- A. Reagent: Tris 150 mmol/L, L-alanine 750 mmol/L, lactate dehydrogenase > 1350 U/L, pH 7.3.
- B. Reagent: NADH 1.9 mmol/L, 2-oxoglutarate 75 mmol/L, Sodium hydroxide 148 mmol/L,sodium azide 9.5 g/L
- WARNING: H302: Harmful if swallowed. EUH031: Contact with acids liberates toxic gas.
- P301+P312: IF SWALLOWED: Call a POISON CENTER or doctor/physician if you feel unwell. P330: Rinse mouth.
- For further warnings and precautions, see the product safety data sheet (SDS).
- Store at 2-8ºC.
- Reagents are stable until the expiry date shown on the label when stored tightly closed and if
contaminations are prevented during their use.
- Indications of deterioration:Reagents: Presence of particulate material, turbidity, absorbance of the blank lower than
1.400 at 340 nm (1 cm cuvette).
- C. Reagent (cod 11666): Pyridoxal phosphate 10 mmol/L. 5 mL.
Biosystems Alaninaminotransferaz (ALT/GPT) REAGENT PREPARATION
- Working Reagent: Pour the contents of the Reagent B into the Reagent A bottle. Mix gently. Other volumes can
be prepared in the proportion: 4 mL Reagent A + 1 mL Reagent B (Note 1). Stable for 1 month at 2-8ºC.
- Working Reagent with Pyridoxal Phosphate (Note 2): Mix as follows: 10 mL of Working Reagent + 0.1 mL of Reagent C (cod 11666). Stable for 6 days at 2-8ºC.
- Analyzer, spectrophotometer or photometer with cell holder thermostatable at 30 or 37ºC and able to read at 340 nm.
- Cuvettes with 1 cm light path.
- Serum and plasma collected by standard procedures.
- Alanine aminotransferase in serum and plasma is stable for 7 days at 2-8ºC. Use heparin or
- EDTA as anticoagulant7