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Alkaline Phosphatase cleaves p-nitrophenyl phosphate (p-NPP) in the presence of magnesium ions, to form p-nitrophenol and phosphate.
p-nitrophenyl phosphate+ h20 p-nitrophenol+phosphate
The intensity of the yellow colour formed is directly proportional to the Alkaline Phosphatase activity in the specimen and is measured kinetically.
Preparation Of Working Solution
Reconstitute the substrate tablet with diluent as per the instruction indicated on the diluent bottle. Swirl gently.
Reagent Storage, Stability & Handling
AutoPure Alkaline Phosphatase is a singal reagent system.
Stable till the expiry date indicated on the label, when stored at 2 8 C
On- Board Reagent Stability
21 days 2- 8 C after opening Protect the reagent from light and contamination.
Do not freeze the reagent
Components & Concentration Of Working Solution
Specimen Collection & Preservation
Collect sample using standard sampling tube. Haemolyzed specimen should be avoided as it may falsely elevate results. Serum or heparinized plasma can be used. Anticogulants suc as EDTA oxalate or citrate inhibit the Alkaline Phosphatase activity and hence cannot be used. Alkaline Phosphatase activity in serum/ plasma is stable for 4 days at 2- 8 C and 28 days at -20 C . Centrifuge samples containing precipitate before performing assay.
Fully automated system automatically calculate the Alkaline Phosphatase activity of each sample.
Result in ukat/l= Result in IU/Ix 0.0167
Refer to application sheet for details. For additional system application, contact our local Accurex representative.
For calibration , it is recommended to use C.f,a,s or any other suitable calibrator material.
Re- calibration is recommended
Absorption of atmospheric CO2 by the opened reagent bottle leads to impaired calibration stability.
Each batch of AutoPure Alkaline Phosphatase is assayed with alteast six quality control sera Precinorm, Precipath, Biorad l, Accutrol Normal and Accutrol Abnormal prior to release.
To ensure adequate control, it is recommended that the labortory should use a normal and abnormal commercial reference control the reagent and instrument functions together.
If the control values fall outside the specified limits, each of the below criteria should be crosee checked and corrected:
Expected range varies from population to population. It is therefore recommended that each laboratory should establish its own normal range. For diagnostic purposes, the alkaline phosphatase results should always be assessed in conjuction with the patients medical history, clinical examinations and other findings.
With AutoPure Alkaline Phosphatase, the assay is linear upto 2000IU/I(33.4ukat/l). Determine samples with higher concentrations via the rerun function. On instruments without rerun function , manually dilute the samples with higher concentration using 0.9%NaCl or distilled/ deionized water Multiply the result with the appropriate dilution factor
There is no significant interference in samples containing upto 25 mg/dl of bilirubin and 400mg/dl of haemoglobin.
Reproducibilty was determined using quality control sera as shown below
Quality control material
low control serum
High control serum