ALKALINE

PHOSPHATASE
Group 6 BSMT 3-A

ALKALINE PHOSPHATASE
EC

Code Number

3.1.3.1

Systematic

Orthophosphoric monoester phosphohydrolase

(alkaline optimum)

Standard

Name

ALP

Abbreviation

ALKALINE PHOSPHATASE
It

is a nonspecific enzyme capable of reacting

with many different substrates.

It

functions to liberate inorganic phosphate from

an organic phosphate ester with the
concomitant production of an alcohol.

In

healthy sera, alkaline phosphatase (ALP)

levels are derived from liver and bone
(osteoblasts).

Bone

isoenzyme increases due to osteoblastic

activity and is normally elevated in children
during periods of growth and in adults older
than age 50 years (geriatric).

ALKALINE PHOSPHATASE

In normal pregnancy, increased ALP activity can be

detected between 16-20 weeks of pregnancy.

The presence of intestinal ALP isoenzyme in serum

depends on the blood group (secretor gene and H
substance) of the individual B or O blood group
increases intestinal ALP after consumption of a fatty
meal.

ALP is also higher in individuals of group B and O than in

A and AB individuals because of differences in intestinal
ALP levels.

Placental ALP is also lower in pregnant women of groups

A and AB.

ALKALINE PHOSPHATASE
Major

tissues sources

liver, bone, placenta and intestinal

Reference
Major

value: 30-90 U/L

Isoenzymes

Liver ALP
Bone ALP
Placental ALP
Intestinal ALP

ALKALINE PHOSPHATASE
Carcinoplacental

ALP

Regan ALP
- is found in lung, breast, ovarian and gynecological
cancers; bone ALP co-migrator; most heat stable
ALP (65 degrees Celsius for 30 minutes); inhibited
by phenylalanine reagent

Nagao ALP
- found in adenocarcinoma of the pancreas and
bile duct, pleural cancer; variant of Regan ALP;
inhibited by L-leucine and phenylalanine.

ALKALINE PHOSPHATASE
METHODS
Electrophoresis

Liver and bone ALPs are the most anodal isoenzymes; intestinal
ALP is the least anodal.
Use of neuraminidase and wheat germ lectin improves
separation of bone and liver ALPs.
High-Resolution electrophoresis using polyacrylamide gel and
isoelectric focusing are capable of resolving multiple bands of
ALP.

Heat Fractionation/Stability Test

It is performed at 56oC for 10-15 minutes.

Placental ALP is the most heat stable; bone ALP is the most heat
labile.
Decreasing order of ALP heat stability: placental, intestinal, liver
and bone

ALKALINE PHOSPHATASE
METHODS
Chemical Inhibition

Bowers and Mc Comb (Szasz Modification)

This method uses different concentrations of phenylalanine,

synthetic urea and levamisole solutions.
Placental and intestinal ALPs are inhibited by phenylalanine
reagent and 3M urea inhibits bone ALP.
Levamisole reagent inhibits liver and bone ALP.

It is considered as the most specific method; IFCC recommended

method.
It is a continuous-monitoring technique which requires a pH
environment of 10.15 and should be read at 405 nm.

ALP
p-nitrophenylphosphate <-----> p-nitrophenol + phosphate ion

ALKALINE PHOSPHATASE
Summary of ALP Methods
METHODS

SUBSTRATE

END PRODUCTS

Beta-glycerolPO4

InorganicPO4 +
Glycerol

5. King and Armstrong

Phenylphosphate

Phenol

6. Bessy, Lowry & Brock

p-nitro phenyl PO 4
(PNPP)

p-nitrophenol or yellow
nitrophenoxide ion

Phenolphthalein diphosphate

Phenolphthalein red

Alpha-naphthol PO 4

Alpha-naphthol

Buffered phenolphthalein PO 4

Free phenolphthalein

1. Bodansky
2. Shinowara
3. Jones
4. Reinhart

7. Bowers and McComb

8. Huggins and Talalay

9. Moss
10. Klein, Babson & Read

ALKALINE PHOSPHATASE
Clinical Significance
Elevations of ALP are of most diagnostic significance
in the evaluation of hepatobiliary and bone
disorders. In hepatobiliary disorders, elevations are
more predominant in obstructive conditions than in
hepatocellular disorders; in bone disorders,
elevations are observed when there is involvement
of osteoblasts.

In biliary tract obstruction, ALP levels range from 3 to

10 times the ULN. Increases are primarily a result of
increased synthesis of the enzyme induced by
cholestasis. In contrast, hepatocellular disorders,
such as hepatitis and cirrhosis, show only slight
increases, usually less than three times the ULN.

ALKALINE PHOSPHATASE
Clinical Significance

Elevated ALP levels may be observed in various bone disorders.

Perhaps the highest elevations of ALP activity occur in Pagets
disease (osteitis deformans). Other bone disorders include
osteomalacia, rickets, hyperparathyroidism, and osteogenic sarcoma.
In addition, increased levels are observed in healing bone fractures
and during periods of physiologic bone growth.

In normal pregnancy, increased ALP activity, averaging approximately

1 times the ULN, can be detected between weeks 16 and 20 is
two to three times the ULN during the third trimester. ALP activity
increases and persists until the onset of labor. Activity then returns
to normal within 3 to 6 days. Elevations also may be seen in
complications of pregnancy such as hypertension, preeclampsia, as
well as threatened abortion.

ALKALINE PHOSPHATASE

ALP levels are significantly decreased in the inherited

condition of hypophostasia. Subnormal activity is a result
of the absence of bone isoenzyme and results in
inadequate bone calcification.

When total ALP levels are increased, it is the major liver

fraction that is most frequently elevated, especially in
obstructive jaundice.

ALP is increased in obstructive jaundice due to greater rate of

secretion.

For bone disorders, highest elevations occur in Pagets disease

(osteitis deformans),

ALKALINE PHOSPHATASE

Increased ALP: Osteitis deformans, Obstructive

jaundice, Osteomalacia, Rickets, Osteoblastic bone
tumors (Osteosarcoma), Sprue, Hyperparathyroidism,
Hepatitis and Cirrhosis (slight increased)

ALKALINE PHOSPHATASE

Important Notes:

Zinc is a component of ALP, and magnesium is the

enzyme activator.

Ingestion of food leads to release of intestinal ALP

into lymphatic fluid, and may transiently increase
plasma levels of ALP.

Hemolysis and diet (fatty meals) sources of

analytical errors; elevated serum ALP.

ALP is sensitive if stored at low temperature (4C)

leads to increased serum level.

ALP is inhibited by phosphorus the addition of 2amino-2methyl-1-propanol (AMP) buffer binds

phosphorus under Bowers-McComb method.

ALKALINE PHOSPHATASE

Important Notes:

Decreased ALP is seen in zinc deficiency.

Transient low serum ALP may occur after blood

transfusion or cardiopulmonary bypass.

Prolonged low levels of ALP occur in hypophosphatasia.

Placental alkaline phosphatase (PLAP) is a useful tumor

marker in serum and cerebrospinal fluid (CSF) for most
germ cell tumors CSF levels of PLAP are of diagnostic
value in differentiating whether a tumor in the pineal body
is a pinealoma or a germ cell tumor.

ALKALINE PHOSPHATASE
Reference Range:
ALP (total)
Males/Females
Males

Females

4-15 y
20-50 y
60 & above
20-50 y
60 & above

54-369 U/L
53-128 U/L
56-199 U/L
42-98 U/L
53-141 U/L

ALKALINE PHOSPHATASE
Sources of Error
hemolysis slight elevations (ALP 6x conc. in
RBCs)
inc. serum activity (3%-10%) = standing at 25
or 4 degrees Celsius
inc. 25% = ingestion of a high-fat meal

ALKALINE PHOSPHATASE
References:

Rodriguez, MT, Clinical Chemistry Review Handbook for

Medical Technology Students (2014)
Bishop, ML et al., Clinical Chemistry Principles, Techniques and
Correlations (7th Edition)

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