Pathological urine biochemistry mgm.pptx

Ppt about urine biochemistry

1. Pathological Constituents of Urine Department of Biochemistry MGMCH

2. contains certain abnormal constituents U r i n e i s sa i d t o be p a t ho l og i c al when it in detectable amounts. That is: Proteins Carbohydrates Ketone bodies Bile salt and pigments Blood

3. 1. Protein Proteinuria: The presence of proteins in urine is known as proteinuria. Proteinuria may be functional or organic.

4. Proteinuria Functional P r o t einur i a Organic P r o t einuria Pre-renal Proteinuria: Fever, heart failure, Temporary & harmless May occur after vigorous exercise, Liver and collagen disease. a cold bath & in pregnancy. Renal Proteinuria: Acute and chronic glomerulonephritis, Nephrosclerosis, renal tuberculosis & carcinoma. Post-renal Proteinuria: Inflammation & lesions of urinary tract.

5. Bence-Jones protein: A peculiar protein Derived from immunoglobulin Seen in multiple myeloma and leukemia Remains dissolved at lower & higher temperature.

6. 2. Carbohydrates G l yc o su ri a : T h e p r e s e n ce of d et ec t a b le amo u n ts of carbohydrates in urine is known as Glycosuria. The common causes of Glycosuria are: Diabetes mellitus Other endocrine disorders such as hyperpitutarism, Cushing’s syndrome & pheochromocytoma. E m o ti o n al o r s t r ess g l y c o su r i a : D u e t o h y p e r s ec r e ti o n of epinephrine under emotional stress. Renal Glycosuria: benign condition in which the capacity of the renal tubules of reabsorb glucose is subnormal. Alimentary Glycosuria: Glucose appears in urine transiently after an excessive ingestion of carbohydrates.

7. 3. Ketone Bodies The ketone bodies include acetoacetate, beta- hydroxybutyrate and acetone. They are formed normally in liver from acetyl CoA. Oxidized in extra hepatic tissues. Ketosis: The excessive production of ketone bodies leading to ketonemia and ketonuria is k/as Ketosis. Ketosis is observed in severe DM, Prolong starvation & toxemia of pregnancy.

8. 4. Bile salts Sod i u m & P o t as s i um sa l t o f g l ucu r on i c & taurocholic acid. Formed in liver & excreted in bile. Excreted in urine during obstructive jaundice. Detection of bile salt in urine sample is significant for differentiation of obstructive & hemolytic jaundice.

9. 5. Blood Hematuria: The presence of intact RBCs in the urine is known Hematuria. Acute glomerulonephritis Cancer of urinary tract Stone in urinary tract Injury to urinary tract Bleeding disorders such as hemophilia, purpura & leukemia.

10. Hemoglobinuria: Hemolytic anemia Typhoid fever Extensive burns Mismatched blood transfusion

11. Tests for Pathological Urine Protein: Heat coagulation test Carbohydrate: Benedict’s test Ketone bodies: Rothera’s test Bile salt: Hay’s sulphur test Blood: Benzidine test

12. Test for Protein: Heat coagulation test Aim: T o p e r f or m h e a t c o a g u l a tion t e s t wi t h gi v en u r i n e sample. Reagent: Chlorophenol red indicator. Acetic acid 1%.

13. Principle: When a protein is heated, its physical, chemical with biological properties are changed due to breaking up of certain bonds and resultant change in conformation of its molecules. This process is called denaturation. However, when the coagulable proteins are heated at their isoelectric pH, a series of changes occur involving dissociation of protein subunits, uncoiling of polypeptide chains, while a denatured protein may be restored to its original structure and function by certain manipulations, coagulation is an irreversible process. Chlorophenol red (CPR) is used to adjust the pH of solution to isoelectric pH of albumin, which is about 5.4. CPR gives a pink color at this pH, a purple red color above it & a yellow color below it.

14. Procedure: Fil l 2 / 3 of t h e t e s t t u b e w ith p r o t e in s ol u tio n . A d d 4 - 5 d r o p s of chlorophenol red & mix. A purple red color develops. Add 1% Acetic acid drop by drop with until the color change to faint pink. Hold the test tube at its bottom incline slightly & heat its upper portion of fluid. A dense coagulation is formed in the upper portion of solution. It may be compared with lower part of solution which serve as a control. This test is given by heat coagulate protein. Eg. Globulin & albumin. Observation Table: R e s ul t : Al bum i n i s h e a t c oag u la b le p r o t e in w h ile G e l a tin, Casei n & Peptone are non-heat coagulable protein. Experiment Observation Inference Perform heat coagulation test with given urine sample Dense coagulation develops at upper portion. Given urine sample contains protein ( A l bu m i n / gl o bu li n ).

15. Test for Carbohydrate: Benedict’s test Aim: To perform Benedicts test with given urine sample Reagent: Benedict’s reagent Principle: CuSO 4 hydrolyzes to form cupric hydroxide which is reduced to cuprous oxide on heating with a reducing carbohydrate. In Benedict test Alkaline medium is provided by the Na 2 CO 3 and precipitation of Cu (OH) 2 is checked by Sodium citrate.

16. Procedure: Take 5 ml of Benedict reagent in a test tube. Add 8 drops of carbohydrate solution and mix it. Boil the mixture for 2 min. The appearance of a green, yellow, orange, or red precipitate indicates that the carbohydrate is a reducing one. Observation Table: Result: Given urine sample contains carbohydrate. Experiment Observation Inference Perform Benedict’s test with given urine sample Orange color appeared Given urine sample contains carbohydrate.

17. Test for Ketone bodies: Rothera’s Test Aim: To perform Rothera’s test with the given urine sample. Reagent: Freshly prepared 5% solution of sodium nitroprusside. Solid ammonium sulphate. Concentrated ammonia. Principle: Ketone bodies react with sodium nitroprusside in the presence of alkaline medium to form permangnate color complex.

18. Procedure: – Take 5 ml of urine sample in a test tube and saturate it with solid ammonium sulphate. o f s o d i u m Add 2-3 drop of a freshly prepared 5% solution nitroprusside. Add 2-3 drops of conc. Ammonia mix and allow to stand. A pp e a r a n c e o f p e r m a n g a n a t e c olor i nd i c a t e s p r es e n c e bodies. o f k e t o n e Observation Table: Result: – Given sample of urine contains Ketone bodies. Experiment Observation Inference Perform Rothera’s test with given sample Permanganate color appear Given sample of urine contains Ketone bodies.

19. Test for Bile salts Aim: To perform Hay’s sulphur test with the given urine sample. Reagent: Sulphur powder. Principle: Bile salt lowers the surface tension of urine (obstructive jaundice) and hence sulphur powder sinks to the bottom. Procedure: T a k e 5 m l o f u ri n e s a m p le i n a t e s t tu b e. S p ri n k l e su l phu r powder on surface. Sulphur powder will start sinking in the bottom due to surface tension.

20. Observation Table: Result: Given sample of urine contains bile salt. Experiment Observation Inference Perform Hay’s sulphur test with given sample of urine Sulphur powder sinks Given sample of urine contains bile salt.

21. Test for blood A im : To sample. p e r f or m B e n z i d i n e t e s t w i th the gi v en u r i n e Reagent: Benzidine solution Hydrogen peroxide Principle: The peroxide activity of heme oxidizes H2O2 to release nascent oxygen which reacts with Benzidine to form bluish green complex. H2O2 + O2 H2O = (O)

22. Procedure: Take 3 ml of urine sample in a test tube and add 1-2 drops of benzidine solution. Mix and add 1-2 drops of H2O2. A bluish green color will appear. Observation Table: Result: Given sample of urine contains blood. Experiment Observation Inference Perform Benzidine test with given sample of urine Bluish green color appears Given sample of urine contains blood.

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