UMA CO., LTD.

MEASURE ALT (SGPT)

2-19-6 Yokosuka Reagent for measuring Alanine aminotransferase
Matsudo, Chiba, Japan JSCC Method

　2 ~ 8 °C IVD In vitro Diagnostics Packages

R1: 90 mL R2: 30 mL
　DO NOT freeze  18 months/block from light R1: 60 mL R2: 20 mL

1. PURPOSE OF USE glutamate.

In vitro Determination of Alanine aminotransferase (ALT) activity Lactate dehydrogenase (LDH) catalyzes the reduction of pyruvate
in serum or plasma. and the simultaneous oxidation of NADH to NAD+. Activity of ALT
can be determined by measuring the rate of decrease of this
2. GENERAL INSTRUCTION
NADH.
1. For in vitro diagnostics use only.
2. Diagnosis should be made in a comprehensive manner, in L-alanine + α – ketoglutarate ALT
→
accordance with other related test results and clinical symptoms
Pyruvate + L-Glutamate
by the doctor in attendance.
3. For guaranteed results, usage of this product must comply with Pyruvate + NADH + H +
LDH L- Lactate + NAD +
+ H2O
→
the instruction in this manual.
4. Follow the automatic analyzers instructions carefully.
7. CALCULATION & UNIT CONVERSION
3. MATERIALS REQUIRED BUT NOT INCLUDED
Calculation
- Saline 0.9 % and high grade purified water
- Calculate ∆Abs of specimen & standards vs blank
- Micropipet and other basic laboratory equipment.
- Plot a calibration curve ALT (U/L) = f(∆Abs)
- Calibrators and Controls (separatedly sold)
- Calculate ALT concentration in specimen using the curve (doing
4. REAGENT COMPOSITION & PREPARATION
same procedure for Controls)
- Reagent R-1: L-alanine; NADH; LDH
Unit conversion
Reagent R-1 is ready for use
1 U/L = 0.017 µkat/L
- Reagent R-2: L-alanine; α-ketoglutaric acid
8. PERFORMANCE & CORRELATION TEST
Reagent R-2 is ready for use
Performance
- Calibrators: K.Factor calibration (Hitachi 7180 K. -4251)
- Sensitivity: Change in absorbance when using purified water is
- Controls Lyo-1 & Lyo-2 (separatedly sold): Put 1 mL of purified
below 0.001 ~ 0.012/min. and change in absorbance using
water to the vials of controls (L, H); leave at room temparature for
solution (ALT 1000 IU/L) as sample is 0.100 ~ 0.300 /min.
30 minutes before use. After reconstituted, controls can be use
- Specificity: The accuracy is within ±10.0%.
without dilution.
- Reproducibility: CV value < 10.0%.
5. SAMPLE PREPARATION & STORAGE
- Measuring range: 3 ~ 1000 U/L.
- Serum: Wait until sample completly coagulated. Take the
Reference Material for Calibration
supernatant to use as specimen.
ReCCS JCCLS CRM-001
- Plasma: Treat sample by anticoagulant; leave sample to stand
9. EXPECTED VALUES
for 3 hours or centrifuge at 2000 rpm for 2 minutes; take the
- Male: 8 ~ 42 U/L
plasma layer (supernatant) and use as specimen.
- Female: 6 ~ 27 U/L
- Analyze sample soon after collection. In case, it could not be
Reference range should be established at each facility and
analyzed soon, store sample 2 ~ 8 C and analyze within 3 days.
judgement should base on measurement results in a
6. MEASUREMENT PRINCIPLE
comprehensive manner together with clinical symptoms and other
ALT catalyzes the transfer of the amino group of L-alanine to α-
measurement results.
ketoglutarate and resulting in the formation of pyruvate and L-

1/2 Revision 11/2013

10. INTERFERENCES 2. Do not use the container and auxiliaries included in this kit for
- Influence of ascorbic acid of 50 mg/dL, conjugated bilirubin of 20 other purposes.
mg/dL, or hemoglobin of 190 mg/dL to the sample of ALT 40 U/L 3. Do not mix reagents of different lot for use.
is within 10%. 4. Do not add to the reagent being used even if it is the same lot
- Avoid to use hemolyzed specimen. number.
Disposal
11. INFORMATION FOR AUTOANALYZERS
1. All specimens, as well as all instruments (e.g. test tubes) that
 For Hitachi Analyzers
come in contact with the specimens, must be treated by the
Calculation Method Rate
following methods, or they must be treated according to the
Temperature 37 C
Specimen 8.5 manual for infectious medical waste provided in each facility.
Volume (μL) R1 150 ・ Sterilize with an autoclave, subjecting them to high pressure
R2 50 saturated steam at 121 °C for more than 20 minutes. Do not
Main 340 process waste containing sodium hypochlorite solution with an
Wavelength (nm)
Sub- 600 autoclave.
Point 1 10
Measurement ・ Immerse at least one hour in sodium hypochlorite solution
Point 2 21
(cycle) (active chloride concentration of over 1000 ppm).
Point 3 34
2. This reagent contains sodium azide. Sodium azide can react
Calibration type K: -4251
with lead pipe and/or steel pipe and can generate explosive metal
Unit U/L
azide. Make sure to use plenty of water at disposal. Concentration
of sodium azide in R-2 is 0.05%.
 For Beckman Coulter Analyzers

Calculation Method Rate 13. OTHER INSTRUCTIONS AND CAUTION

Temperature 37 C - Results may differ depending on the sample/reagent ratio.
Specimen 8.5 Adjust parameters for different analyzer.
Volume (μL) R1 150 - Prepare the calibration curve on the day of determination.
R2 50
Main 340
Wavelength (nm)
Sub- 600
Point 1 14
Measurement Point 2 27
(cycle) Point 3 -
Point 4 -
Calibration type Linear
Unit U/L

12. HANDLING, USAGE & DISPOSAL

Handling
1. Specimen can be potentially positive for infectious agents
including hepatitis B virus and HIV. Wear glove and goggle when
needed.2. In case reagents got into skin, eye or mouth by mistake,
wash it immediately with plenty of water and consult the doctor if
needed.
3. If reagents are spilled, dilute with water and wipe it out. If
specimen is spilled, spray 80% of alcohol over the specimen and
wipe it out.
Usage
1. Store reagents under specified condition. Do not use after
expiration date.

2/2 Revision 11/2013