Bansil, Noemi Jean Cabrega, Maria Kathrinna Macuha, Nina Christine 4DMT Liver Profile Test (Liver Function

Test) LIVER It is reddish brown and located in the upper right quadrant of the abdomen under the diaphragm. It is the largest and most complex organ of the gastro intestinal tract. Its structural unit is called lobule. GROSS ANATOMY The liver weighs approximately 1.2-1.5 kg in a normal healthy adult. It is divided unequally into two lobes by the falciform ligament, with the right lobe being the approximately six times larger than the left lobe. FUNCTIONS I. Metabolic function A. Bilirubin metabolism Bilirubin is the major metabolite of heme. Production of bilirubin is about 250 350 mg daily in normal individual. B. Amino acid metabolism Ammonia is the product of urea cycle in the amino acid metabolism Increase ammonia can result from Reyes syndrome or acute hepatic failure. C. Lipid metabolism The liver is the major site of metabolism for cholesterol, apoproteins and all lipoprotein except chylomicrons. D. Bile acids They are derived from cholesterol. It facilitates fats in the intestines. II. Synthetic function The liver is responsible for the formation of the following substance: A. Proteins albumin, and globulin, enzymes B. Carbohydrates glucose and glycogen C. Lipids phospholipids, lipoproteins, cholesterol, high and low density lipoprotein. D. Ceruloplasmin it catalyzes the conversion of iron to ferric state. E. Plasma clotting factors III. Excretory/ Secretory function A. Bile Primary substance excreted by the liver. Made up of bile acids or salts, bile pigments, cholesterol and other substance extracted from the blood. Stored temporarily in the gall bladder.

B. Bilirubin Principal pigment in bile. IV. Detoxifying function Liver serves as gatekeeper between substances absorbed by the gastrointestinal tract and those released in systemic circulation. Most important mechanism of the liver is the drug metabolizing system. Liver Profile Is a biochemical determination to assess liver function Liver Diseases I. Jaundice - yellow discoloration of the eyes, skin, and mucous membranes most often resulting from the retention of bilirubin. - bilirubin levels reach3.0 mg/dL A. Prehepatic Jaundice - occurs when the problem is prior to liver metabolism. - commonly caused by increased amount of red blood cell destruction. - B1 is increased; B2 is normal B. Hepatic Jaundice - occurs when the primary problem resides in the liver. - B1 and B2 is increased. Gilberts Syndrome - a benign hereditary disorder in the transport of bilirubin in the sinusoidal membrane of the hepatocyte. Criggler-Najjar Syndrome - an inherited disorder of bilirubin metabolism that causes defective conjugation due to the deficiency of the enzyme uridine diphosphate glucoronyl transferase (UDP) Dubin-Johnson Syndrome - an inborn error that causes elevated serum levels of conjugated bilirubin due to a blockade of the excretion of bilirubin into the canaliculi. Rotor Syndrome - this condition is of viral origin. It is characterized by impaired excretion of conjugated bilirubin. C. Posthepatic Jaundice - results from biliary obstructive disease that prevents the flow of conjugated bilirubin into the bile canaliculi. - B2 is increased. Kernicterus disorder of the central nervous system in incidences of hyperbilirubinemia among infants attributed to premature Blood Brain Barrier. II. Hepatitis A. Acute Hepatitis There is an inflammatory response in the liver which is lymphocytic due to ischemia or toxins. Hallmark of acute hepatic injury is elevation of aminotransferase levels. B. Chronic hepatitis It is a major predisposing factor for cirrhosis and hepatocellular carcinoma. III. Cirrhosis It is a clinical condition in which scar tissue replaces normal liver tissue. Irreversible

IV. Reyes syndrome Describes a group of disorder caused by infectious, metabolic, toxic, or drug induced disease found mostly in children. V. Tumors Hepatocellular carcinoma or hepatoma associated with previous infection with hepatitis virus. Reference intervals for Liver Function Test Laboratory test Reference intervals Albumin 3.8 5.0 (38-50 g/L) Ammonia <120 ug/dL (67 umol/L) Bile acids 0.3 -3.0 mg/dL (3.0 -30 mg/L) Bilirubin Direct <0.3 mg/dL (<5 umol/L) Indirect 0.1 - 1.0 mg/dL (2 17 umol/L) Total 0.1 1.2 mg/dL (2 21 umol/L) Ceruloplasmin 23 50 mg/dL (230 500 mg/L) Prothrombin time 11 15 secs Liver Tests Metabolic, Excretory and Secretory Function Tests BILIRUBIN DETERMINATION o TOTAL BILIRUBIN Is determined using indirect Van den Burgh method that uses diazotized sulfanilic acid (DSA) solution on the serum with the addition of a dissociating agent to form a red azo dye called azobilirubin Different dissociating agents: o 50% Methanol Malloy and Evelyn Procedure o Caffeine Sodium Benzoate Jendrassik-Groff Metod for Total and Conjugated Bilirubin Determination o DIRECT BILIRUBIN Is determined using direct Van den Burgh method that also uses diazotized sulfanilic acid (DSA) solution on the serum but with no addition of any dissociating agent o Only Total and Conjugated (direct) Bilirubin are measured. Unconjugated (indirect) Bilirubin is determined by subtracting conjugated bilirubin from total bilirubin. o Other test: urine bilirubin determination Bromsulphalein (BSP) o obsolete test for liver function (hepatic excretory capacity) in which a known amount of dye, usually 5 mg/kg of body weight, is injected intravenously, usually after 45 minutes, the amount of dye remaining in the serum is measured. o A concentration of 0.4 mg or less of bromsulphalein per 100 mL of serum or less than 4% of the injected dye is considered normal o Bromsulphalein retention may follow decreased hepatic blood flow or biliary obstruction as well as hepatic cell damage.

Metabolic Function Test CARBOHYDRATE METABOLISM o GLUCOSE TOLERANCE TEST is a laboratory method to check how the body breaks down (metabolizes) sugar requires fasting for 8-10 hours for the fasting blood sugar the patient will be asked to a liquid containing a certain amount of glucose then blood will be extracted after an hour and after another hour o other tests include: Galactose Tolerance Test Fructose Tolerance Test Lactic Acid Tolerance Test Glucagon Tolerance Test Epinephrine Tolerance Test

Detoxification and Synthetic Function Tests Detoxification Quicks Test/ Hippuric Acid Test o The patient is given at test meal, Sodium Benzoate which will be conjugated with glycine to form non-toxic hippuric acid that is excreted in urine if the liver is normally functioning. Synthetic Almost all proteins are synthesized except immunoglobulins Plasma clotting factors o Most are synthesized except for von Willebrands factor. Prothrombin time o Increased in patients on warfarin therapy, vitamin K deficient, those with acute hepatitis, and hepatocellular carcinoma. Protein Metabolism I. Total Protein Albumin/Globulin II. Flocculation and Turbidity test a. Cephalin cholesterol Flocculation tests b. Thymol Turbidity Test c. Zinc Sulfate Turbidity Test Enzymology -The amount of enzymes assess the extent of damage of an organ Alkaline Phosphatase (ALP) It belongs to the group of enzymes that catalyze the hydrolysis of various phosphomonoesters at an alkaline pH. It functions to liberate inorganic phosphate from an organic phosphate ester with the production of an alcohol. Increased in hepatocellular disease but markedly elevated in obstructive diseases. Lab Methods: 1. Bessey- Lowry- Brock: p-nitrophenylphosphate

2. Bowers- McComb: p-nitrophenylphosphate 3. King- Armstrong: phenylphosphate 4. Sinowara- Jones- Reinhart: - glycerophosphate Gamma Glutamyl Transferase (GGT) It is the transfer of gamma glutamyl group from a gamma glutamyl peptide to another peptide or amino acid. Increases after alcohol intake Lab Method: Photometric enzyme reaction (Szasz assay) Lactate Dehydrogenase (LD) It is the oxidation of L-lactate to pyruvate with the mediation of NAD or hydrogen acceptor. Primary fraction that is associated with the liver Lab Methods: 1. Wacker method: utilizes lactate to pyruvate reaction with formation of NADH to NAD 2. Wroblewski LaDue method: employs the reverse reaction- pyruvate to lactate Alanine Aminotransferase (ALT) It catalyzes the transfer of an amino acid from alanine to -ketoglutarate with the formation of glutamate and pyruvate. It is distributed in many tissues, with higher concentration in the liver. It is considered as more liver specific enzyme of the transferases. Lab Methods: 1. Coupled Enzyme reaction 2. Reaction with dinitrophenyhydrazone (DNPH): Reitman- Frankel 3. Coupling with diazonium salts Aspartate Aminotransferase (AST) It is involved in the transfer of an amino group in aspartic acid for an -keto group in ketoglutaric acid forming oxaloacetate and glutamate. The highest concentrations are found in the liver, cardiac and skeletal muscles. Lab Methods: 1. Karmen Method: incorporates a coupled enzymatic reaction using MD as indicator 2. Reaction with DNPH: keto acids fromed ate oxidized by tricarboxylic acid cycle 3. Coupling with diazonium salts: ketoacid + diazo compound diazonium derivative

References: Bishop, M.L., Fody, E.P., and Schoeff, L.E.(2010). Clinical Chemistry: Techniques, Principles, Correlation. 6th ed. Lippincott Williams & Wilkins. Philadelphia. U.S.A McPherson, R. &Pincus, M. (2006).Henrys Clinical Diagnosis and Management by Laboratory Methods.US : Saunders Elsevier http://www.medilexicon.com/medicaldictionary.php?t=90423 http://www.nlm.nih.gov/medlineplus/ency/article/003466.htm