Alkaline Phosphatase Flashcards Preview

Clinical Chemistry Laboratory 2 > Alkaline Phosphatase > Flashcards

Flashcards in Alkaline Phosphatase Deck (19)
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0

Method

Liquicolor, Colorimetric test

1

Reaction principle

p-Nitrophenylphosphate + water -- (AP) -- phosphate + p-nitrophenol

2

Contents

1 Reagent 1/Buffer solution: Diethanolamine buffer (pH 9.8)
2 Reagent 2/Substrate solution: p-Nitrophenylphosphate

3

Specimen

1 Serum
2 Heparinized plasma

4

Loss of activity within 7 days at 4 Celsius

0%

5

Loss of activity at 20-25 Celsius

10%

6

Procedure 1 with reagent start

The reagents are stable, even after opening, up to the stated expiry date when stored at 2-8 Celsius. Contamination of the reagents must be avoided.

7

Procedure 2 with sample start

The working reagent is stable for 4 weeks at 2-8 Celsius and for 5 days at 15-25 Celsius

8

Wavelength

Hg 405 nm (400-420 nm)

9

Optical path

1 cm

10

Temperature

1 25 Celsius
2 30 Celsius
3 37 Celsius

11

T or F. Read against air (increasing absorbance)

T

12

Procedure with sample start

Mix sample and reagent mixture, read the absorbance after 1 minute and at the same time, start the stop watch. Read the absorbance again exactly after 1, 2, and 3 minutes.

13

Linearity

If the absorbance change per minute exceeds 0.250, dilute 0.1 mL of the sample with 0.5 mL physiological saline (0.9%) and repeat the assay using this solution. Multiply the result by 6.

14

Calculation

1 Method 1 with reagent start
U/l = mean absorbance per minute x 3433
2 Method 2 with sample start
U/l = mean absorbance per minute x 2757

15

Conversion factor

(Review)

16

Reference range

(Review)

17

Contain sodium azide as preservative

1 Buffer solution (R1)
2 Substrate solution (R2)

18

Produced during the reaction

p-nitrophenol