LIVER FUNCTION TEST: BOARD EXAM

12mg

How many mg of albumin is excreted daily

Serum albumin and Vitamin K coag factors

Tests the measures the synthetic ability of the liver

Total protein

•Marker for nutritional status

•Also, it measures the function of different organs such as liver, kidney and bone marrow

•Usually performed on non fasting serum

•Hemolysis and Icteric sample can falsely elevate this test especially in biuret method

0.2 to 0.4mg/dl

Plasma levels of total protein is _____ higher than serum because of fibrinogen

Transudates

Total protein is less than 3mg/dl

Exudates

Total protein is greater than 3mg/dl

Kjeldahl method

It is the reference method for total protein measurement but it is not routinely used.

•measures the nitrogen content of protein (1g of nitrogen is equivalent to 6.54g of protein)

•uses tungstic acid to produce protein free filtrate

•

H2so4

Is the digesting agent used in kjelfahl method

Ammonia

End product of kjedahl method

Biuret method

Most widely used in cc according to ifcc. And it used 2 peptide bonds and alkaline medium to measure total protein

Principle of biuret method

Cupric ions involved in the peptide bond forming violet colored compound which is proportional to number of peptide bonds present and reflects total protein level at 545nm

•Alkaline copper sulfate

•Rochelle salt (NaK tartrate)

•NaOH and Potassium iodide

Reagent used in biuret method

Folin-Ciocalteu (Lowry) Method

Highest analytical sensitivity that involves the oxidation of phenolic compounds such as tyrosine, tryptophan and histidine to give blue color compound

Phosphotungstic-molybdic acid or phenol reagent

Main reagent used in folin ciocalteu method

Biuret Reagent

Color developer used in folin ciocalteu method

Ultraviolet absorption method

Absorbance in 210 nm due to peptide bonds at specific wavelength

Absorbance at 280nm

Due to tryptophan, tyrosine and phenlyalanine

SPE

Involves the migration of charge particles. Identification of monoclonal spike of Ig and differentiating them from polyclonal hypergammaglobulinemia

Elevated aat, haptoglobin and a1-antichymotrypsin

Myocardial infarction

Albumin

Fastest band in SPE

Crp and Immunoglobulins

Gamma globulin

Transferrin, beta-lipoprotein, hemopexin and complement

Beta-globulin

Haptoglobin, AMG, Ceruloplasmin

Alpha2 globulin

Glycoprotein, AAT, AAG, Thyroxine binding globulin,

Alpha1 globulin

Alpha1-antitypsin

Decreased in emphysema and juvenile hepatic cirrhosis

Multiple myeloma

Gamma spike

Hepatic cirrhosis

Beta gamma bridging

Alpha2 globulin band spike

Nephrotic syndrome

Alpha1 globulin flat curve

Juvenile cirrhosis

Spikes at alpha1, alpha2 and beta globulin bands

Inflammation

Presence of hemoglobin

Causes a "blip" in late alpha2 and early beta zone

iron deficiency anemia (transferrin)

Presence of small spikes in beta region

Rheumatoid arthritis and Malignancy

Causes polyclonal gammopathy

Refractometry

Alternative test for chemical analysis of proteins

Turbidimetric and Nephelometric Methods

Utilize sulfosalicylic acid or trichloroacetic acid

Albumin

Soluble in water and concentrated salt solution

Globulin

Soluble in weak salt soln and hydrocarbon solvents

Coomasie Brilliant Blue

Detects the proteins to as little as 1ug

Ninhydrin

Develops violet color that reacts with primary amines. Widely used after paper chromatography, amino acid analysis and thin layer chromatography

Malignancy, Multiple myeloma and waldenstrom macroglobulinemia

Increase in total protein

Prothrombin time or Vitamin K Response test

2nd test for liver function. It differentiates intrahepatic which is prolonged protime and extrahepatic which is normal protime.

•administrated intramuscularly, 10mg for 1 to 3days

Bcg

Most widely used dye

Bcp

Most specific dye

Bcg

Affected by presence of penicillin, falsely low protein is the result

HABA

Affected by icteric sample

Bcg and bcp

Not affected by hemolysis and icteric sample

Artifactual hyperalbuminemia

Caused by prolonged tourniquet application

Cirrhosis, Hepatitis and Obstructive jaundice

•Increased or normal in Total protein

•Decrease albumin

•Increased globulin

Nephrosis and malabsorption

•Decreased total protein

•Decreased albumin

•Normal globulin

Salt retention

Decrease in TPAG

Dehydration

Increased TPAG

Multiple myeloma

•Increased total protein and globulin

•Normal albumin

Analbuminemia

Hereditary absence of albumin

Bisalbuminemia

Presence of 2 albumin bands due to penicillin drug

Cirrhosis, Multiple myeloma and waldenstrom macroglob

Diseases that usually observe in inverted albumin/globumin ratio.

B1

Marker for conjugation function

B2

Marker for excretion function

B1

Indirect reacting, hemobilirubin,slow reacting, prehepatic bilirubin

Delta bilirubin

Conjugated bilirubin tightly bound to albumin.

•commonly seen in patients with obstructive jaundice and can be used as monitoring test for gallstone removal.

•not calculated on neonatal px less than 14 days old

Jaundice

Bilirubin levels exceeds mg/dl

Pre-hepatic jaundice

Increased in indirect bilirubin and caused by too much destruction of Rbc.

•Increased B1 while normal B2

•Urobilinogen is normal while negative for bilirubin in urine

•Malaria, HDN, Hemolytic anemia

Post hepatic jaundice

Increased to B2 while B1 is normal

•Urobilinogen is Decreased while positive bilirubin in urine

•Caused by cholelithiasis and pancreatic tumor

Hepatocellular combined jaundice

Both increase B1 and B2

•Urobilinogen is decreased while positive bilirubin in urine

Alp

Enzyme test for post hepatic jaundice.

Gilbert's syndrome

Bilirubin transport deficit

•impaired cellular uptake of bilirubin

•mild kernicterus

•elevated b1

Type 1 Crigler-Najjar Syndrome

Deficiency in enzyme UDPGT

•total absence of b2 production

Type 2 Crigler-Najjar syndrome

Partial deficiency in UDPGT

•slight b2 are produced

Dubin-Johnson syndrome and Rotor syndrome

Bilirubin excretion deficit.

•increased in b2 and total bilirubin

Lucey-Driscoll Syndrome

Presence of inhibitor for bilirubin conjugation

•elevated b1

Placenta normally removes bilirubin

Why does fetus with hemolytic disease does not develop hyperbilirubinemia

450nm

In liley test, bilirubin in amniotic fluid is absorbed at what specific wavelength

Unbound or free bilirubin

What causes the blood brain barrier resulting to kernicterus

Elevated

In panhepatic cirrhosis, the indirect and direct bilirubin is.

Falsely increase bilirubin

Hemolysis can cause

Falsely decrease

Lipemic sample can cause

Exceeds 25mg/dl

Visible icterisia occurs when

Caffeine and methanol

This is the accelerant for measurement of unconjugated bilirubin

Conjugated or direct reacting bilirubin

This will be measured if accelerants are elliminated

Unconjugated bilirubin (B1)

Bilirubin standard solution is made up of what?

Bilirubin assay

Diazonization of bilirubin to produce azobilirubin

Evelyn and malloy method

Uses

Diazo A= 0.1%sulfanilic acid and HCl

Diazo B= 0.5% sodium nitrite

Diazo blank= 1.5% hcl

Methanol

Coupling accelerator in evelyn and malloy method

Pink to purple azobilirubin

End product of evelyn and malloy method for bilirubin assay

Jendrassik and grof

This is the ph dependent method for bilirubin. Most commonly used for discreet analyzer.

•not affected by hemoglobin up to 750mg/dl and ph changes.

Diazo reagent

Main reagent in jendragroff

Sodium acetate

Buffer used in jendrassik grof

Ascorbic acid

This is a composition in jendrassik and grof method for bilirubin assay that terminates the initial reaction and destroys the excess diazo reagent

Alkaline tartrate solution

Composition of jendrassik grof method that provides alkaine ph

Pink to blue azobilirubin

End product of jendrassik and grof method

Conjugated bilirubin

What type of bilirubin will produce color in acqeous soln

Unconjugated bilirubin

What type of bilirubin will produce a color after the addition of an alcohol

15 mins

Total bilirubin is being measured after how many minutes after the addition of methanol or caffeine solution

Caffeine benzoate

It does not causes precipitation of proteins and increased turbidity compared to methanol

Solubilization of unconjugated bilirubin before chemical quantitation

Measurement of total bilirubin involves what?

BSP Dye excretion test

Test that evaluates the hepatocellular function and potency of bile duct

Rosenthal white (double collection method)

Dose= 2mg/kg body weight of the patient. Specimen is collected into 2 manner, after 5 mins and after 30 mins

Mcdonald method (single collection)

Dose= 5mg/kg body weight and specimen is collected after 45 mins

Urobilinogen

Colorless end product of bilirubin metabolism

2 hour freshly collected urine or freshly collected stool

Specimen for urobilinogen detection

Ehrlich's method

Method for urobilinogen detection