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Khadija  Balubaid KAU-Faculty of Science- Biochemistry department Khadija  Balubaid KAU-Faculty of Science- Biochemistry department

Khadija Balubaid KAU-Faculty of Science- Biochemistry department - PowerPoint Presentation

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Khadija Balubaid KAU-Faculty of Science- Biochemistry department - PPT Presentation

Clinical biochemistry lab BIOC 416 2013 Liver Function profile LFT Enzymes Liver Liver is an important organ in human body Synthesis of proteins glycogen storage drug metabolism and detoxification process ID: 934041

serum liver alt ast liver serum ast alt disease normal due damage alp reagent injury hepatitis heart muscle high

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Slide1

Khadija BalubaidKAU-Faculty of Science- Biochemistry department Clinical biochemistry lab (BIOC 416)2013

Liver Function profile (LFT)Enzymes

Slide2

Liver Liver is an important organ in human body Synthesis of proteins, glycogen storage, drug metabolism and detoxification processMany diseases can affect liver functions as:

Viruses (heptites A,B,C,D,G)

Cirrhosis Inflammation JaundiceFatty liver

Slide3

Slide4

Slide5

Liver profile Profile : is group of tests specific for one organ Liver function tests (LFTs or LFs), are groups of clinical biochemistry laboratory

blood (serum or plasma) or urine assays designed to give information about the state of a patient's liver

These tests can be used to detect the presence of liver disease

distinguish

among different types of liver disorders

Gauge

the extent of known liver damage,

follow

the response to treatment.

Slide6

Slide7

SGPT (Serum Glutamate Pyruvate Transaminase) Liver enzymes

Produced by liver cells.

ALT found primarily in liver.Normal : 7 – 41 U/LUpto 300U/L – nonspecific , any type of liver disorder(CLD…cirrhosis /malignancy)>1000U/L – extensive hepatocellular damage ( viral hepatitis, ischemic liver injury , toxin /drug induced liver injury )

ALT (alanine amino

transferase

) or

Slide8

High serum ALT due to: Liver cells damage due to inflammation, virus infection or cell death (why?) when liver cells damaged ALT enzyme leaks to blood stream leads to rise its level in serum. Some medication may also elevate serum ALT, because some drugs cause liver damage leads to rise ALT level.

ALT is the most sensitive marker for liver cell damage;

since it is only synthesized by liver cells other enzymes may be also synthesized by other organs.

Slide9

Less sensitive that ALTSynthesized by : liver, cardiac muscle, skeletal muscles, kidneys, brain, pancreas, lungs,

leucocytes and RBC Normal – 12 – 38U/L

AST – liver , cardiac muscles, skeletal muscle, kidneys, brain, pancreas, lungs, leucocytes, RBC in decreasing order.2 Iso enzymes- cytoplasmic, mitochondrialMild degree of tissue injury – cytoplasmic form in serum

Severe injury – mitochondrial type in serum

AST (

Aspartate

amino

transferase

)

or

SGOT (Serum Glutamate

Oxaloacetate

Transaminase

)

Slide10

High serum AST due to: Muscle damage, myocardial infarction (heart attack) and in chronic liver disease. To confirm that high AST is due to heart or muscle injury; other enzyme (creatinine kinase CK) which is specific for heart, is also tested.

Because it is less sensitive the ration ALT/AST is calculated ALT

: < 35U/L, AST: <40U/L elevated ALT,AST : acute hepatitis (viral or toxic ), chronic hepatitis and cirrhosis,biliary obstruction

Slide11

ALP (alkaline phosphatase)It is related to bile duct.ALP

normal level: 30-130 U/L It is not specific for bile

because it is synthesized also by bone and placenta (isoenzymes)High serum ALP may be due to: bile duct

damage

(inflammation, cirrhosis or obstruction)

In alcohol hepatitis

.

Hepatocellular carcinoma

Slide12

Normal physiological elevation :During pregnancy During child growth

To assess the etiology of ALP elevation, GGT and bilirubin levels are also measured.

Slide13

Used in body for syn of glutathione11 iso enzymesNormal : 9 – 58

U/LProduced by liver, kidney and

pancreas, intestinal cells,and prostateElevated in: toxins, alcoholism,obstructive

Jaundice,and

neoplasm of liver

Slightly high normally in males

prostate

To

detect alcohol abuse

Rised

even when other LFT are normal in

alcohalics

.

GGT falls rapidly within few days after abstinence

.

Used To

confirm hepatic etiology of ALP elevation

GGT (Gamma

Glutamic

Transpeptidase

)

Slide14

General indications Conformational procedures Indications Enzymes Serology (for virus)Biopsy, ultrasound (liver size)Hepatitis. may be due to ( virus, medication, toxin)

ALTAST/ALT ratio, CK to confirm heart disease Not specific.

May be due to ( muscle disease, heart disease, liver disease) ASTGGT to confirm liver Gallbladder ultrasound Bile problems (stone or bilary duct obstruction) Liver disease Normal physiological elevation ( child, pregnancy)

ALP

Liver toxin, alcohol,

cirohsis

GGT

In general, every enzyme can gives you specific indication:

Slide15

Experiment:Measuring serum AST levelPrinciple: The rate of NADH oxidation is directly related to AST activity which measured photometrically. L-aspartate +

a oxoglutarate L-glutamate +

oxaloacetateOxaloacetate + NADH + H+

L-

malate

+ NAD

+

MDH

AST

Slide16

Notes:Samples: Unhemolyzed serum or plasma collected in heparin or EDTA tube.(why?)

Stability of AST in serum: 2 days at 20-25o

C or 4 days at 2-8oC

Slide17

Kit components Reagent 1: mixture of: buffer (pH 7.5) +

substrate (L-aspartate)

Reagent 2: mixture of: enzyme (MDH) + coenzyme (NADH)

Slide18

Procedure Prepare working reagent: by mixing reagent 1 and 2 together Zero adjust the spectrophotometer with air or dis. H2O

Prepare the reaction as the following:

Mix After 30 sec. read the absorbance at 340nm. (R1) Repeat the reading after 1min and 2 min (R2, R3)

Calculate the mean absorbance

mean = R1+R2+R3/ 3 or (R2-R1)+(R3-R2)/2

Sample tube

1 ml

Working reagent

100

m

l

Sample (serum)

Slide19

Calculations:25oC

18 U/L Men

15 U/L Women

AST catalytic conc. U/L = mean A X factor

* factor = 1946

Reference value “ normal rang”:

Slide20

Interfering factors Therapeutic heparin increase ASTHemolysed blood increase ASTMany drugs falsely increase or decrease AST