University of Santo Tomas – Legazpi

Clinical Chemistry 2 Laboratory (MT 114.1)

Name: Trisha O. Garcia

Block: 3BSMT1

Alkaline Phosphatase Determination

Answer the following:

What is the optimum pH needed to allow ALP determination to proceed?

How is it different from ACP?

The pH environment is the primary distinction between ACP and ALP determination. The optimal pH
range for determining ALP is between pH 9 and pH 10. As the name suggests, ACP determination
requires an acidic environment medium, ideally at 5.0 pH.

Give two non-pathologic conditions where ALP level is elevated.

Normal pregnancy. Elevated ALP activity can be noticed between 16-20 weeks of pregnancy.
Additionally, it is 2-3 times of upper limit of normal during the trimester. ALP rises during pregnancy
and remains until labor starts.

Physiologic bone growth. Bone isoenzymes increases due to osteoblastic activity and is normally
elevated in children during growth spurt and in geriatric patients who are older than 50 years of age.

Enumerate the tissue sources of ALP.

The tissue sources of ALP are the bone, liver, placenta, and intestines.

Describe briefly the methods for ALP isoenzymes differentiation.

Electrophoresis. Differences in electric charges cause protein molecules to separate.

Neuraminidase and wheat germ lectin are used to separate the two ALP isoenzymes as it is
challenging to tell bone and liver ALPs apart from one another. Liver, bone, placental, and
intestinal appear in decreasing order of ALP isoenzyme charges.

Heat Fractionation/Stability Test. This test is performed at 56 degrees Celsius for 10 minutes. The
decreasing order of ALP isoenzymes heat stability: placental, intestinal, liver, and bone. This test is
also used in distinguishing between the bone and liver isoenzyme. There are two outcomes after
heating: (1) If the remaining activity is <20% after heating, the bone is the source of ALP elevation;
and (2) If the remaining activity is >20% after heating, the liver is the source of ALP elevation.

Chemical Inhibition Test. This method utilizes different chemicals to determine the source of ALP
elevation. This is mostly used in the determination of intestinal and placental ALP through the
inhibitory action brought upon phenylalanine. If the action is inhibited at 75%, the intestine (intestinal
ALP) is the source of ALP elevation. On the other hand, if the action is inhibited by 80%, the placenta
(placental ALP) is the source of ALP elevation.
To what condition is ALP most indicated?
University of Santo Tomas – Legazpi
Clinical Chemistry 2 Laboratory (MT 114.1)

Bone disorders and liver problem are the conditions where ALP is most indicated. Bone disorders
such as Paget’s disease, bone tumors, and rickets increased bone ALP. Liver problems such as
blocked bile duct, cirrhosis, and hepatitis.

Give the laboratory methods for ALP with their corresponding substrates.

METHOD SUBSTRATE
Bodansky
Shinowara
b-glycerol phosphate
Jones
Reinhart
King and Armstrong Phenylphosphate
Bessy, Lowry, & Brock P-nitrophenyl phosphate (PNPP)
Bowers and McComb P-nitrophenyl phosphate (PNPP)
Huggins and Talalay Phenolphthalein diphosphate
Moss a-naphthol phosphate
Klein, Babson, & Read Buffered phenolphthalein phosphate