2 x 12.

5, 2 x 35 mL, 2 x 70 mL
ALKALINE PHOSPHATASE (S.L) IFCC
11401008, 11401009, 11401010

Intended Use
Waste Management
This reagent is intended for in vitro quantitative determination of Al kaline
Reagent must be disposed off in accordance with local regulation.
Phosphatase in serum or plasma.
- Kinetic Method recommended by IFCC Sample
- Linear up to 2000 U/L Serum/Plasma (free of haemolysis).
- Working reagent can be prepared as per requirement
- Pack sizes suit to all types of laboratories Interferences
Clinical Significance No interference for
Alkaline phosphatase (ALP) is widely distributed throughout the body, but clinically Haemoglobin up to 1000 mg/dL
important one for diagnostic reasons are in bone, liver, placenta and intestine. Growing Bilirubin up to 40 mg/dL
bone is associated with the release of ALP and so in child hood the level of ALP is Ascorbic Acid up to 50 mg/dL
around 3 times of that of adult. During pregnancy in 2nd and 3rdtrimester the enzyme
rises considerably due to placenta releasing ALP. It can be used to examine placental Materials Provided
function.
Al kaline Phosphatase R1 & Al kaline Phosphatase R2.
Elevated levels are seen in bone diseases, e.g. Paget’s disease, Rickets, Osteoblastic
metastatic and in obstructive disease of biliary tract. Materials required but not provided
Decreased levels are rarely seen. e.g. in Vitamin A resistant rickets.  Pipettes& Tips
 Test Tubes & racks
Principle  Timer
Al kaline phosphatase acts in highly al kaline pH in presence of divalent Mg ions  Incubator
where it catalyses the hydrolysis of p-Nitrolphenylphosphate (PNPP) which results
 Analyzer
in release of p-Nitrophenol and free phosphate group. Absorbance is proportional
to the serum alkaline phosphatase at 405 nm.
The enzyme al kaline phosphatase hydrolizes the 4-NPP to release 4-nitrophenol, Test Parameters
under alkaline conditions. The 4-nitrophenol formed is detected spectrophotometrically Mode of Reaction Kinetic
at 405 nm to give a measurement of al kaline phosphatase activity in the sample.
Slope of reaction Increasing
ALP Wavelength 405 nm
Para-nitrophenyl phosphate + H2O p-nitrophenol + inorganic phosphate
++
pH 10.2,Mg
Temperature 37 C
0

Factor 2757
Kit Components
Blank DI water
Reagent/ Product code Product code Product code Description Linearity 2000 U/L
Component 11401008 11401009 11401010
Delay time 60 sec
Al kaline 2 x 10 mL 2 x 28 mL 2 x 56 mL HEDTA 8.78g/L No of readings 3
Phosphatase Zink sulfate 6mL/L
(S.L) R1 Interval 60 sec
Magnesium acetate-5g/L
IFCC Sample volume 20 µL
AMP buffer 92.6
Reagent volume 1000 µL
Al kaline 2 x 2.5 mL 2 x 7 mL 2 x 14 mL P-Nitro phenylphosphate Cuvette 1 cm light path
Phosphatase -50 mmol /L
( S.L) R2
Application parameters for various instrument are available. Please contact
customer support department for specific information.
Risk & safety
Material safety data sheets (MSDS) will be provided on request. Calibration
Reagent Preparation Agappe multicalibrator is recommended for calibration of this assay on fully
auto analyzers.
Mix 4 volume of Reagent 1 (R1) with 1 volume of Reagent 2 (R2)
Use provided factor (2757) for estimation of ALP semi auto analyzers.
The working reagent is stable for 30 days at 2-80C.
Note : Discard the working reagent if the blank absorbance exceeds 1.00 at 405 Procedure notes
nm.
Laboratory Procedure for Semi Auto
Reagent Storage and Stability
The sealed reagents are stable up to the expiry date stated on the label, when Working reagent 1000 µL
0
stored at 2-8 C and protected from light. Sample 20 µL
0
Open Vial Stability Mix and incubate at 37 C for one minute. Measure the change in absorbance
o
Once opened the reagent is stable up to 4 weeks at 2- 8 C, if contamination is per minute ( OD/min) during 3 minutes.
avoided
Onboard Calibration Stability Calculation
Onboard calibration stability is 7 days. ALP Activity (U/L) = (OD / min.) x 2757.

Reagent Deterioration Quality control

Turbidity or precipitation in any kit component indicates deterioration and the It is recommended to use Agappe Qualicheck Norm and Path (11601001) to verify
component must be discarded. Values outside the recommended acceptable range the performance of the assay. Each laboratory has to establish its own internal
for the Agappe control may also be an indication of reagent instability and quality control scheme and procedure for corrective action, if control do not recover
associated results are invalid. Sample should be retested using a fresh vial of within the acceptable range.
reagents.
Precaution
To avoid contamination, use clean laboratory wares. Avoid direct exposure of
working reagent to light.

‘Agappe Hills’, Dist. Ernakulam, Kerala, India-683 562.

Tel. +91 484 2867 000 | Customer Support No.: 1800 425 7151(Toll Free) ISO 9001 : 2015
customersupport@agappe.in | www.agappe.com REV. NO.: ADL/IFU/ALPIFCC/LIQ/R02 EN ISO 13485: 2016
 2 x 12.5, 2 x 35 mL, 2 x 70 mL
ALKALINE PHOSPHATASE (S.L) IFCC
11401008, 11401009, 11401010

Reference Range
It is recommended that each laboratory establish its own reference values. The Inter Run
following value may be used as guide line. Control Level 1 Control Level 2
Women : 35 - 104 U/L n 20 20
Men : 40 - 129 U/L Mean (U/L) 96.13 355.8
SD 3.26 15.65
Children up to15 yrs : 54 - 369 U/L
CV(%) 3.39 4.4
Results obtained for patient samples are to be correlated with clinical finding of
patient for interpretation and diagnosis.
Accuracy ( U/L )
Performance Control Expected Value Measured Value
1. Linearity
Control Level 1 103 ± 28 102.08
The reagent is linear, up to 2000 U/L.
If the concentration is greater than linearity (2000 U/L), dilute the sample Control Level 2 390 ± 67 376.58
with normal saline & repeat the assay. Multiply the result with dilution factor.
2. Comparison Sensitivity
A comparison study has been performed between Agappe reagent and another Lower detection Limit is 6.0 U/L
internationally available reagent yielded a correlation coefficient of r2= 0.9949
and a regression equation of y =0.9545x. Bibliography
3. Precision 1. Clin.Chim.Acta,(1983)339 F-367 F
2. Tietz Textbook of Clinical Chemistry
Intra Run 3. IFCC Primary Reference Procedure for the Measurement of Ctalytic Activity
Control Level 1 Control Level 2 Concentrations of Enzymes at 37 0C; Part 9. Reference Procedure for the
n 20 20 measurement of Catalytic Concentration of Alakline Phosphatase
Mean (U/L) 96.26 361.66 4. Jose L. Mil lan, Purinergic Signal ling (2006) 2: 335Y341; Al kaline
SD 0.86 1.99 Phosphatases Structure, substrate specificity and functional relatedness to
CV(%) 0.89 0.55 other members of a large superfamily of enzymes

‘Agappe Hills’, Dist. Ernakulam, Kerala, India-683 562.

Tel. +91 484 2867 000 | Customer Support No.: 1800 425 7151(Toll Free) ISO 9001 : 2015
customersupport@agappe.in | www.agappe.com REV. NO.: ADL/IFU/ALPIFCC/LIQ/R02 EN ISO 13485: 2016