ALKALINE PHOSPHATASE-FLUID (5+1) DGKC IVD

Optimized Determination of Alkaline Phosphatase in Body Fluids (DGKC Method)

MEASUREMENT PRINCIPLE TEST COMPARISION

AP Comparision with another commercial available test on the Hitachi
p-Nitro-phenylphosphate + H20 → p-Nitrophenolate 717:
+ Phosphate n = 58; r = 0.9986; y = 0.9983x – 1.7311

The p-Nitrophenol is present as yellow coloured p-Nitrophenolate. ASSAY PROCEDURE

The increase of absorbance per minute at 405 nm is proportional to
the enzyme activity. Wavelength : 405 nm
Temperature : 37° C
CONCENTRATION OF WORKING SOLUTION
Buffer Stability of reaction mixture:
Diethanolamin (pH=10.0) 0.6 mol/1 at + 2° - + 8°C: 25 days
MgCl2 0.4 mmol/1 at +18° - +22°C: 5 days
Starter
p-Nitrophenylphosphate 7 mmol/1 Pipette into cuvette:
Start with serum
STORAGE AND STABILITY Reaction mixture (only for start with serum): Mix reagent 1 (5
The sealed reagents are stable up to the indicated expiry date if volumes) with reagent 2 (1 volume) and temperate before use.
stored at +2° - +8°C.
Reaction mixture 1000 µl
INTENDED USE Sample 20 µl
In vitro test for the quantitative determination of Alkaline
Phosphatase in human serum and plasma. Start with R2:
Buffer R1 1000 µl
PRECAUTIONS AND WARNINGS Sample 20 µl
For in vitro diagnostic use only. Attend to the normal precautions Reagent 2 200 µl
required for handling all laboratory reagents. Buffer reagent contain
Diethanolamin. In the case of contact rinse affected area with plenty Mix and incubate for 30 sec. and measure the changes of
of water; get immediate medical attention after contact with eyes or absorbance for at least 3 min.
if ingested. Determine the differences in absorbance/min. (δA/min)

EXPECTED VALUES CALCULATION

( in serum, Heparin plasma ) at 37 °C
Factor for serum start
U/l µkat/1 F x δA/min (U/l) 2757
Children: 180 - 1200 1.3 - 20 F x δA/min (µkat/l) 45,9
Adults: male: 100 - 270 1.67 – 4.50
female: 100 – 240 1.67 – 4.00 Factor for substrate start R2
F x δA/min (U/l) 3298
SAMPLE MATERIAL F x δA/min (µkat/l) 54,9
Serum, Heparin Plasma. Hemolysis will interfere.
The values in serum controls increase directly after reconstitution: APPLICATIONS
Measure controls after reconstitution after 1 hour at +20°C Applications are available for many analyzers after request

QUALITY CONTROL LITERATURE

Centronorm, Centropath or all other control sera with values Thomas,L.:Labor u.Diagnostik,l.Aufl.,S.64,
determinated by this method may be employed. Med.Verlag,Marburg,(1978)
Weishaar,D.,Grossrau,E. u.Faderl,B.: Mediz.Welt 26, 5.387-
LINEARITY 390,(1975)
935 U/l
At higher values repeat the test using a sample diluted 1 + 9 with PACKAGE SIZES
0.9% NaCI. Multiply the result by 10.

CONVERSION FACTOR KIT SIZE ORDER NO.

U/l µkat/l Factor: 0.0167 2 x 60 ml manual AF02000060
µkat/l U/l Factor: 60 1 x 120 ml manual AF02R05060
4 x 120 ml manual AF02000120
10 x 60 ml for Hitachi AF02911060
TEST SPECIFICATIONS 704/902/717/911
Reproducibility in Intraassay and Interassay on Hitachi 717: 5 x 120 ml for Hitachi 717/911 AF02911120
5 x 120 ml for Hitachi 911 barc. AF02911120-B
Within run (n= 30) Between run (n= 30) 6 x 84 ml for Hitachi 917/ AF02917084-B
Mean SD Mean SD Modular-P
Sample %CV %CV 6 x 84 ml for Olympus AU AF02917084
U/l U/l U/l U/l
8 x 30 ml for Selectra AF02SEL030
Level 1 70.8 1.54 2.18
Level 2 277.4 2.75 0.99 276 4.84 1.75
Centronic GmbH Am Kleinfeld 11,
Level 3 525 6.01 1.14 484.7 12.68 2.62
85456 Wartenberg/Germany
Phone: 0049-8762724300, Fax: 0049-8762724312
Edition 04/2013, TV-004-CE-003 e-mail: [email protected]
web: www.centronic-gmbh.com