Alkaline phosphatase (ALP)

Alkaline phosphatase is a hydrolase enzyme responsible for removing phosphate groups

from many types of molecules, including nucleotides, proteins, and alkaloids. The
process of removing the phosphate group is called dephosphorylation.

It occurs in osteoblasts, hepatocytes, the kidneys, spleen, placenta, prostate, leukocytes

and small intestine.the liver-bone-kidney type is particularly important.

Physiology

In humans, alkaline phosphatase is present in all tissues throughout the entire body, but is
particularly concentrated in liver ,bile duct, kidney, bone, and the placenta. The optimal
pH for the enzyme activity is pH=10 in standard conditions.
The enzyme is termed alkaline phosphatase because it works under alkaline (non-acidic)
conditions, as opposed to acid phosphatase

Why It Is Done

A test for alkaline phosphatase (ALP) is done to:

• Check for liver disease or damage to the liver. Symptoms of liver disease can include
jaundice, belly pain, nausea, and vomiting. An ALP test may also be used to check the
liver when drugs that can damage the liver are taken.

• Check bone problems (sometimes found on X-rays), such as rickets, osteomalacia,

bone tumors, Paget's disease, or too much of the hormone that controls bone growth
(parathyroid hormone). The ALP level can be used to check how well treatment for
Paget's disease is working.

• Check the cause of a high blood calcium level.

 Results

• An alkaline phosphatase (ALP) test measures the amount of the enzyme ALP in the
blood.

Normal

Normal values may vary from lab to lab.

Alkaline phosphatase
Adults: 30–126 units per liter (U/L)
Children: 30–300 U/L

Women in the third trimester of pregnancy have high ALP levels because the placenta
makes ALP. Children normally have much higher ALP than adults because rapid bone
growth is normal in children and bones make ALP.

High ALP levels can show that the bile ducts are blocked.Levels are significantly higher
in children and pregnant women.

High values
Very high levels of ALP can be caused by liver problems, such as:

• Hepatitis
• Blockage of the bile ducts (obstructive jaundice),
• Gallstones,
• Cirrhosis,
• Liver cancer, or
• Cancer that has spread (metastasized) to the liver from another part of the body.

High ALP levels can be caused by bone diseases, such as:

• Paget's disease,
• Osteomalacia,
• Rickets,
• Bone tumors,
• Tumors that have spread from another part of the body to the bone
• Hyperparathyroidism
• Normal healing of a bone fracture can also raise ALP levels.
Rare Causes:

• Heart failure,
• Heart attack,
• Infectious mononucleosis
• KIdney cancer
• Sepsis

Low values

A decreased serum alkaline phosphatase may be due to:

• Zinc deficiency.
• Hypothyroidism.
• Vitamin C deficiency/Scurvy.
• Folic acid deficiency.
• Excess Vitamin D intake.
• Low phosphorus levels (hypophosphatasia)
• Celiac disease.
• Malnutrition with low protein assimilation (including low stomach acid
production/hypochlorhydria).
• Insufficient Parathyroid gland function.
• Pernicious anemia
• Vitamin B6 insufficiency

What Affects the Test?

Reasons you may not be able to have the test or why the results may not be helpful
include:

• Taking medicines that may damage the liver, such as some antibiotics, birth
control pills, long-term aspirin use, and oral diabetes medicines.

• Being pregnant. Women in the third trimester of pregnancy have high ALP levels
because the placenta makes ALP.

• Going through menopause. Postmenopausal women may have higher ALP levels
than women who still have menstrual cycles.

• Your age. Children normally have much higher ALP levels than adults because
rapid bone growth is normal in children and bones make ALP.
 • Using alcohol.

What to Think About

• If the alkaline phosphatase level is high, a special test will help to find the cause.
This test is called alkaline phosphatase isoenzymes, or ALP isoenzymes.

• The isoenzyme profile of alkaline phosphatases can be determined to see if the

elevation of alkaline phosphatase came primarily from liver (ALP-1), bone (ALP-2), or
elsewhere.

• If liver disease is suspected, more blood tests, an ultrasound, or a CT scan are

generally recommended to find the problem.

• Other tests to check liver function, such as alanine aminotransferase, aspartate

aminotransferase, and bilirubin, are often done at the same time as an alkaline
phosphatase (ALP) test. For more information, see the medical tests Alanine
Aminotransferase (ALT), Aspartate Aminotransferase (AST), and Bilirubin.

• Gamma glutamyl transferase (GGT), or gamma glutamyl transpeptidase, may be

measured in the blood to check the difference between bone ALP and liver ALP. High
levels of GGT are present when the liver is damaged but not present with bone disease.
A high level of GGT may be caused by alcohol use or may mean that blocked bile ducts
are causing inflammation. The level of GGT may be high with the use of certain
medicines, such as phenytoin and phenobarbital. In some medical centers, a test that
measures a substance called 5-nucleotidase is done instead of the GGT test because it is
better at finding liver disease.
 SERUM ALKALINE PHOSPHATE ESTIMATION

PRINCIPLE:

Colorimetric assay in accordance with a standardized method.

ALP
P-Nitrophenylphosphate + H2O ---------------- Phosphate + p-Nitrophenol
Mg2+

In the presence of magnesium and zinc ions, p-nitrophenyl phosphate is hydrolyzed by

phopsphatases to form phosphate and p-nitrophenol. The p-nitrophenol released is
proportional to the ALP activity and can be measured photometrically.

REAGENT:

R1:
 Diethanolamine buffer, pH 9.8
 Magnesium sulfate
 Detergents and stabilizers

R2:
 p-nitrophenylphosphate

PREPARATION:

5 parts of R1 are mixed with one part R2. This gives working reagent.

SPECIMEN:

 Collect serum using standard sampling tubes.

 Heparinized plasma.
 Centrifuge samples containing precipitate before performing assay.

TESTING PROCEDURE:

Materials provided:
 Working solution as described above

Additional materials required:

 Calibrators and controls as indicated below
 0.9% NaCl
MANUAL PROCEDURE:

Wave-length: 405nm(400-420nm)
Temperature: +250C/ +300C/ +370C
Cuvette: 1cm light path
Zero adjustment: air or distilled water

Semi micro
Working reagent: 1000µl
Sample: 20µl

Mix and wait for 30 seconds. Read absorbance and start stopwatch simultaneously. Read
again absorbance after exactly 1, 2 and 3 minutes. Calculate ∆A/min

CALCULATION:

1cm cuvette, 405nm ∆A/min × 2757 = ALP(U/l)

A1 = ___________________________

A2 = ___________________________

A3 = ___________________________

Factor = 2757

∆A = A1 + A2+ A3 x factor = ___________________________

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