Safety precautions
For in vitro diagnostic use only.
Do not pipette by mouth.
Exercise the normal precautions required for handling
laboratory reagents.
QUALITY CONTROL
Normal and abnormal control sera of known concentrations
of ALT (SGPT) should be analyzed routinely with each
group of unknown samples
ALT (GPT)
Colorimetric End point
*Reaction principle
*Procedure
ALT (SGPT)
L-Alanine + α-ketoglutarate
Pyruvate +
Glutamate
H+
pyruvate + 2,4 - DNPH-ine
Pyruvate + 2,
4-DNPH-one
The method used here is a modification of the classical
Reitman Frankel colorimetric endpoint reaction.4 In this
procedure ALT (SGPT) catalyzes L-alanine and αketoglutarate to form pyruvate and glutamate. The
pyruvate is then reacted with 2, 4-dinitrophemylhydrazine (2,4-DNPH-ine) to form 2,4-DNPH-one. The
addition of sodium hydroxide dissolves this complex,
allows 2, 4-DNPH-one to be measured at 505
*Reagents composition, Contents and Safety warnings
R1
R2
Wavelength
Optical path
Incubation temperature
Measurement against
546 nm (530 - 550 nm)
1 cm
37°C
Reagent blank
Blank
Absorbance
U/l
Absorbance
U/l
0.025
4
0.275
48
0.050
8
0.300
52
0.075
12
0.325
57
0.100
17
0.350
62
0.125
21
0.375
67
0.150
25
0.400
72
0.175
29
0.425
77
0.200
34
0.450
83
0.225
39
0.475
88
0.250
43
0.500
94
Specimen
Specimen
-----
0.1 ml
Reagent 1
0.5 ml
0.5 ml
Dist. Water
0.1 ml
-----
Mix, incubate for exactly 30 minutes at 37°C.
100 mmol/l
DL-Alanine
80 mmol/l
Reagent 2
Oxo-glutarate
4.0 mmol/l
Mix, allow standing for exactly 20 minutes at 20-25°C.
2-4-dinitrophenyl
hydrazine
4.0 mmol/l
NaOH (0.4 mol/l)
Sample
This assay is intended for use with serum. Reports indicate
that ALT (SGPT) in serum remains stable at 4 - 8°C for a
minimum of seven (7) days.5 Hemolyzed specimens should
not be used as erythrocytes contain fifteen times the ALT
(SGPT) activity in serum.4
Obtain the activity of GPT in the serum from the table
Pipette into test tubes
Phosphate buffer ph 7.4
Storage and Stability of Reagents
Store ALT (SGPT) Substrate and Color Reagent at 2 - 8°C.
All the reagents are stable until the expiration date stated
on the label...
Reagent Preparation
All reagents are ready for use.
*Calculation
0.5 ml
5.0 ml
0.5 ml
Quality Control
Normal and abnormal control sera of known
concentrations of ALT (SGPT) should be analyzed
routinely with each group of unknown samples
5.0 ml
Mix, read the absorbance of specimen (Aspecimen)
against the reagent blank after 5 minutes. The color
intensity is stable for 1 hour.
Bibliography
F. Schiele: Additional information regarding the
recommended method for determining the catalytic
concentration of alanine aminotransferase in human serum
at 30 ° C.: Ann. Biol. Clin. 40, 87 (198 2)
For in vitro diagnostic use only.
The following symbols are used on labels
*EXPECTED VALUES
For in vitro diagnostic use
Use by (last day of the month)
Serum
Up to 12 U/l
Temperature limitation
Batch Code