Liver enzymes

Liver function tests

Liver function Liver function tests rationale

Synthetic function - TPAG total protein albumin Test measuring hepatic synthetic function because liver produce proteins, lipoproteins, clotting
- PT prothrombin time because of the clotting factors enzymes, carbohydrates.
factors

Conjugation and excretion
Detoxification
- Liver is a gatekeeper because
it determines if it can be
released or not.
- Bilirubin
- Urobilinogen
- Enzymes like ALP, aminotransferases, 5’N,
GGT, LDH
- Ammonia; protein catabolism result or
outcome and it’s very toxic that’s why it’s
converted to urea (non-toxic and can easily be
eliminated via kidneys)
Increase ammonia in the blood means
problem in the liver
- OCT (liver marker but not commonly
performed)
- Aminotransferases : AST and ALT
the ability of the liver to metabolized bilirubin
Used to assess the extent of liver damage
- any injury of the liver can lead to liberation of enzyme
Cytolysis and necrosis
Differentiate hepatocellular from obstructive disease
- hepatocellular means that the problem is the hepatocytes (cells from the liver) and
pertain to functional disorder
- obstructive is mechanical, it may be in the form of gallstones, tumor

No storage function test

Methods Substrate End products OTHER METHODS USED IN ALP DETERMINATION

OBodansky Beta-glycerophosphate Inorganic PO4 + glycerol
Shinowara Same Same
Liver enzyme
Jones Characteristics
Same Same Clinical significance Methods of determination Laboratory Considerations
Reinhart - Catalyzes theSame
same catalytic reaction 1. hepatic disorders
Same o Karmen method 1. Serum or heparinized
ALTKing
alanine
and Armstrong with AST where it transfer an amino
Phenylphosphate InPhenol
almost all of the liver diseases ALT plasma free from
aminotransferase
Bessy, Lowry, and Brock group p-nitro
the onlyphenyl
difference
PO4 is that it’s will
p-nitrophenol also
orelevate.
yellow hemolysis
alanine while in AST is aspartate nitrophenoxide: acute
↑ levels ion inflammatory
OldBowers
name SGPT
and McComb - Cofactor: pyridoxal
Same phosphate (co- condition
Same of the liver 2. Aminotransferases are
serum glutamic factor in amino transferases) present in human plasma,
Huggins Talalay Phenolpthalein diphosphate Phenolphthalein red 1st reaction is the catalytic reaction of the
pyruvic - Major tissue source: liver (liver specific ↑ levels of ALT and AST are seen in bile, CSF and saliva
Moss Alpha napthol PO4 Alpha-naphtol PO4 ALT by which catalyzes the transfer of amino
transaminase enzyme) hepatocellular disorders compared - Running enzyme test for
Klein, babson & read Buffered phenolphthalein Free phenolphthalein group from alanine to a-ketoglutarate
- Minor tissue source: kidney pancreas, to extra/intra hepatic obstruction aminotransferases (ALT,
PO4 producing pyruvate and glutamate
EC 2.6.1.2 RBC, heart, lungs and skeletal muscle - Remain elevated for up to 2-6 AST, Amylase) mask
 weeks 2nd reaction the oxidation reduction of NADH should be wear to avoid
Reference value: - Easily detectable to NAD will now be measure. false elevation
6-37 U/L AST/ SGOT ALT/ SGPT o The increase in absorbance of the 3. Vitamin B6
Major organ Heart Liver 2. used to screen blood donors oxidation of NADH to NAD is directly - Vit B6 deficiency patient
affected  screen blood donors; not proportional to the activity of the will have falsely low or
Substrate Aspartic alpha Alanine alpha practiced in the Philippines ALT normal ALT because Vit.
ketoglutaric katoglutaric  usually analyzed are the o 340 nm B6 is the pyridoxal
acid acid transfusion transmittable o 37°C phosphate which is the
End Glutamic Glutamic infections such as HIV, Hepa cofactor
products acid+ acid+ pyruvic C & A, syphilis and malaria  Reitman- Frankel - if absence of cofactor thus
oxaloacetic acid  In US ALT is included in Coupled enzymatic reaction slow enzyme reaction
acid screening for the blood 4. Certain drugs and alcohol
Color 2,4 DNPH Same donors as mandated by the may also increase ALT
developer IMCC because ALT is a good activity
Color 0.4N NaOH Same marker for post-transfusion 5. Stability of the enzyme
intensifier hepatitis; if high ALT si donor, activity can be maintained
Methods Reitman and Reitman and thus not qualified by refrigeration of the
frankel, frankel o the change of the absorbance of sample up to 3 days and
karmen freezing the sample for up
colored complex (brown) is to be
measured to 30 days
o The activity of the colored complex is
directly proportional to the ALT
activity.
o 500 nm

Liver Characteristics Clinical significance Methods of determination Laboratory Considerations

enzyme
Catalytic reaction: liberate 1. Evaluation of hepatobiliary disorder 1. Electrophoresis (cathode-anode) - Hemolysis
ALP the inorganic phosphate ion  Disorder either in the liver, gallbladder, bile - Liver (fastest) , bone, placenta, and intestine - Serum or heparinized plasma less
Alkaline from the organic duct (slowest) than 3 hours old
phos phosphomonoester via  in obstruction: high ALP - Most useful but liver and bone are way too  The more mag stand, the
phatase hydrolysis producing alcohol  Run together with other enzymes to close, thus difficult to differentiate more mag increase ang pH
EC 3.1.3.1  the catalytic differentiate of its hepatic or fsorder.  just add lectin (wheat germ) or the more ma active si ALP
reaction Neuraminidase to improve the - Anticoagulants that remove calcium
happens in 2. Evaluation of bone disorder (Pagets disease) separation of liver and bone and magnesium will prevent produc
 pH 9-10 MAJOR ISOENZYMES 2. Heat fractional/stability test formation
that’s why - Liver ALP - 56 C for 10 mins  False low
alkaline o Found in biliary duct - Placenta (most heat stable because it can - Lipids, hemoglobin or bilirubin
- Nonspecific enzyme - Bone ALP resist denaturation up to 65 up to 30 mins C) interference
- It can react with a o It takes part in the transport of - intestine, liver and; These can be also absorbed light
number of substrate calcium - bone ( unstable; labile first one to denature in @405 nm
- Activator: o Found in osteoclast; in 56C for 10 mins) - Fasting
magnesium children/older; ALP is higher bc of 3. Chemical inhibition test Ideal if fasting because in intestinal
- ALP contains zinc osteoclastic activity - Phenylalanine – inhibit placenta isoenzyme ALP isoenzyme, after a meal mo
- Major: intestine, - Placental ALP and intestine isoenzyme, and raegan & nagao increase. Thus, false increase
liver, bone, placenta o Detected in pregnant women at the same time.Most common isoenzyme - Temp sensitive
- Isoenzymes are starting from 16-20 weeks after - Synthetic urea –is used to inhibit bone At 4°C false increase ALP
named kung aha sila pregnancy then persist all isoenzyme - Zinc deficiency
nagkita nga organ throughout the pregnancy and will - Levamisole –inhibit liver and bone isoenzyme  False low
normalize 3-6 days after nanganak 4. Bower and Mc Comb Hypophosphatasia –genetic disorde
- Intestinal ALP wherein there’s abnormal
o Dependent on the blood group B/O development in the bones and
has higher concentration of - Reaction is based on the breakdown of para- teeth. Low ALP
intestinal ALP than A/AB. Same with nitrophenylphosphate (colorless) because of Low ALP is also found in patients
placental ALP hydrolysis that liberates p-nitrophenol after blood transfusion
o Also involved in the transfer of lipids (yellow) and phosphate ion The ALP tests can be inhibited by
Carcino-placental isoenzyme - p-nitrophenol (yellow) the absorbance is phosphorus
- Carcino because these isoenzymes occur measured at 405 nm Additional information: Placental ALP is a
when there’s neoplasm (active cell division  the increase in absorbance of the good tumor marker for germ cell tumor. If
common in cancer) like: liberated p-nitrophenol directly patient is pregnant it’s expected that high
 Regan- lung, breast ovarian, and colon CA proportional is to the ALT activity placental ALP.
 Nagao –pancreas and bile duct CA - pH 10.15
metastatic CA

Liver enzyme Characteristics Clinical significance Methods of determination Laboratory Considerations

GGT Gamma - Catalytic reaction:
Glutamyltransferase Transfers itself to
amino acids, other
EC 2.3.2.2 peptides or water
molecules
Reference value: - Distribution: kidney,
brain, phosphate,
1. evaluation of liver damage; 1. Szass, Rosalki & Tarrow,
2. detection of alcoholism; and Orlowski  Gamay ra because
3. the monitoring of alcohol intake by patients o Substrate: gamma- GGT is stable
- If nag abstain si patient, then GGT will normalize glutamyl-p-nitroanilide  Not affected by
after 2-3 weeks o Reaction: when the hemolysis
- In all hepatobiliary disorder, GGT will increase; enzyme acted upon the  Can be refrigerated
most sensitive liver assay. It may be obstruction substrate it will convert @ 4°C
Male : 6-45 U/L pancreas, and liver (higher elevation) / functional to glycylglycine that
Female: 5-30 U/L - For the patient that’s taking enzyme inducing liberates p-nitroaniline
- Low because female drugs such as phenytoin, barbital, and warfarin. o Product: p-nitroaniline
has estrogen and Will lead to increase in GGT not because of liver o Chromogenic product
progesterone that damage but bc of induction of GGT o Absorbance will be
may suppress the  GGT man gud is canaliculi of the liver cells measured at 405 – 420
activity of GGT particularly in the smooth ER. Any nm
mitochondrial induction, will trigger the o Method of testing: end-
GGT’s activity that will lead to false point (only one), fixed
increase time, kinetic or
- GGT is also elevated in Acute Pancreatitis, diabetes continuous (several
mellitus, prostatic disorder absorbance reading)
- In AMI, GGT will also increase for unknown reason
- It is also useful in differentiating the cause of ALP
increase:
ALP GGT
Liver Increase Increase
Bone Increase Decrease

5’ Nucleotidase - A phosphoric 1. Hepatobiliary disease - Dixon and Purdon,

monoester hydrolase
EC 3.1.3.5 - Predominantly
secreted in the liver
Reference value: 0-1.6 U/L
2. Also a marker for infiltrative lesions of the liver Campbell, Goldberg
- Also increase in cholestasis. Na stuck or minimized
production of the bile
- Also help to differentiate between the liver and
bone problem in ALP. Just as the same with GGT

Other clinically Characteristics Clinical significance Isoenzymes Laboratory Considerations

significant
enzymes
ACP Acid - Catalyze same reaction as ALP but active at - Significant levels found in: RBC and Band 1 (B1) - Free from hemolysis because B3 is
phosphatase pH 5 platelet. In adult men, 50% found in - found in found in RBC
prostate gland prostate - Decreases when left at Room
EC 3.1.3.2 Methods Substrate End products - One of its diagnostic value is for Band 2 (B2) Temperature
Gutman Phenyl PO4 Inorganic PO4 detecting prostatic cancer or the - found in WBC - If left at RT for 1-2 hrs., decrease
and recurrence of the Prostatic CA particularly in because the pH will increase thus,
↑ in : thrombocytopenia, problems in the granulocytes became an alkaline pH and ACP is
 gutman bone associated with osteoclastic activity, Band 3 (B3) not physiologically active in
Shinowar PNPP p-nitrophenol hemolytic anemias - Found in PLT, alkaline pH
a - Aid in detecting: RBC and - Serum should be frozen or acidify
Babson, Alpha naphthyl Alpha-naphthol  metastatic CA monocytes - Prostatic ACP (B1) is inhibited by L-
Read & PO4 (best if  other types of CA Band 4 (B4) tartrate ions
Phillips kinetic)  bone disease - same as B2 - RBC ACP (B3) is inhibited by
Roy and Thymolphthalein Free  forensic (rape cases) Band 5 (B5) formaldehyde
Hillman monophosphate thymolphthalein Because it’s also found in semen. It - found in - TRAP : tartrate resistance ACP so
(best if end- can persist after 4 days after vaginal osteoclast not affected by inhibitions
point) washing, so detectable Present in people having leukemia,
- Nonspecific enzyme, different lymphomas
substrate Elevated serum bilirubin can cause a
falsely low TRAP

MINOR ENZYMES
Aldolase Catalytic reaction: Splits fructose
- EC 4.1.1.13 Three isoenzymes
ALD A –Skeletal muscle
ALD B –WBC , liver, kidney
ALD C –brain tissue
- Clinical significance: increase in problems in skeletal muscle, hepatic diseases, hemolytic anemia, leukemia

Pseudocholinesterase MAJORITY is in Liver and also liver myocardial endo-pancreas

EC 3.1.1.8 - But not a liver enzyme because it reflects more on the synthetic function rather than the detoxification function (assessed by the liver enzyme).
More of a marker for insecticide poisoning
Decrease in acute hepatitis, cirrhosis, carcinoma & malnutrition
ACE - Convert angiotensin I to angiotensin II
Angiotensin-converting - Monitoring and diagnosis of sarcoidosis: abnormal collection of inflammatory cells
enzyme - EC 3.4.15.1
Ceruplasmin  Marker for Wilson’s disease: genetic disorder wherein there’s excess storage of copper that ables to lodge to the eyes and brain
 Copper carrying protein
Orthinine Carbamoyl - Marker for hepatobiliary diseases
Transferase
G-6-PD o Catalytic reaction: it functions to maintain/ suppress the activity of NADPH in the RBC
o Part of newborn screening
EC 1.1.1.49 o If failure to produce G-6-PD enzyme there might be a problem in the RBC, spleen, adrenal cortex and lymph nodes
o G-6-PD deficiency will lead to hemolysis
o Increase in AMI and megaloblastic anemia