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Normal constituents of urine
- Urine is normally pale yellow and has an aromatic odor. It contains water, salts, urea, creatinine and other substances as it acts to remove waste from the body. - Common tests are used to check for and identify normal constituents like urea, creatinine, uric acid and chloride, as well as check physical properties like specific gravity, pH and volume. - Abnormal results can provide diagnostic information about conditions affecting the kidneys, liver or other body systems based on changes in the levels of these substances excreted in the urine.
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Normal constituents of urine
- 1. NORMAL CONSTITUENTS OF URINE Dr.Janice D’Sa
- 2. INTRODUCTION • Urine isan excretory product of the body. • It is formed in the kidney. • Urine examination helps in the diagnosis of various renal as well as systemic diseases.
- 3. URINE COLLECTION • Urineis usually collected in a sterile wide mouthed container. • Different method of urine collection are – – FIRST MORNING SAMPLE (conc urine for biochemical analysis, casts and crystals) – RANDOM SAMPLE(chemical screening, microscopic examination) – 24 HOUR URINE SAMPLE (quantitative estimation of proteins, sugars, electrolytes, hormones) – MID STREAM URINE – CLEAN CATCH URINE
- 4. SAMPLE PRESERVATION • Fordetermination of urea, ammonia, nitrogen and calcium – Hydrochloric acid is used. • For determination of sodium, potassium, chloride, bicarbonate, calcium, phosphorus, urea, ammonia, amino acids, creatinine, proteins, reducing substances and ketone bodies - Thymol is used • For determination of Ascorbic acid – Acetic acid is used • Toluene may also be used as a preservative.
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- 8. VOLUME • Normal volumeof urine excreted per day by normal subjects is 1000-2000 ml/day. • Night urine output < 400ml. • Factors which influence the volume excreted : – Intake of fluid, proteins and salt. – Excessive perspiration and strenuous exercise decrease the volume of urine.
- 9. COLOR • Normal urineis pale yellow (due to presence of pigment urochrome) • When the output of urine is low it appears deep yellow. • Freshly voided urine is clear and transparent. • Long standing urine may become turbid. (Due to precipitation of phosphates)
- 11. ODOUR • Normal odourof urine is aromatic in nature. • Long standing urine may have an ammoniacal odour (due to decomposition of urea to ammonia).
- 12. PH • Normal PHof urine ranges from 5.5 – 6.5. • Factors which influence PH : – High protein diet makes the urine more acidic – Diet rich in vegetables and fruits make the urine more alkaline. • Measured using Dipsticks, Litmus Paper or Ph paper.
- 14. SPECIFIC GRAVITY • Specificgravity indicates the concentrating ability of the kidneys. • Normal range : 1.012 – 1.024 • Measured using urinometer, refractometer, dipsticks. • Sp. Gravity is affected by – 1. Volume of Urine excreted 2. Amount of solids present in the urine.
- 15. • Urinometer/ Hydrometer consistsof a thin stem graduated from 1000 to 1060 corresponding to specific gravities of 1.0 to 1.06. The bulb at the bottom is suitably weighed. Urinometer is calibrated at 60 deg F (15 deg C)
- 16. METHOD OF DETERMINATIONOF SPECIFIC GRAVITY 1. Take sufficient urine in a container. 2. Allow the urinometer to float in urine without touching the sides. 3. Observe the specific gravity reading corresponding to the meniscus of urine. 4. Note the temperature of urine using a thermometer. 5. Temperature correction is done for specific gravity.
- 17. • TEMPERATURE CORRECTION Observedsp. Gravity = 1010 Observed Room Temperature = 37 deg C Temp of Urine = 15 deg C Corrected Temperature = 37-15/3 = 7 Calibrated Sp. Gravity = 1010 + 7 = 1017 = 1.017
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- 21. CHLORIDE • Normal chlorideexcreted in 24 hr urine sample is 8 – 15 g/ day(NaCl). INCREASED LEVELS DECREASED LEVELS POLYDIPSIA EXCESSIVE SWEATING, FASTING, DIARRHOEA, VOMITING USE OF DIURETICS DIABETES INSIPIDUS ADDISON’S DISEASE INFECTIONS CUSHING’S SYNDROME
- 22. • TEST FORCHLORIDE METHOD – Take 2 ml of urine + 0.5 ml of conc. HNO3 + 1ml AgNo3. OBSERVATION – Curdy white precipitate INFERENCE – Chloride is precipitated as AgCl with Ag No3 in presence of HNO3.
- 23. SULPHATE • Daily excretion: 1g/ day • Sulphur is derived from catabolism of sulphur containing amino acids. INCREASED LEVELS DECREASED LEVELS HOMOCYSTEINURIA RENAL IMPAIRMENT HIGH PROTEIN DIET CYSTINURIA
- 24. METHOD – 2ml urine+ 2ml Barium Chloride OBSERVATION – White precipitate INFERENCE – Sulphate is precipitated as barium sulphate with barium chloride.
- 25. CALCIUM • Daily excretion– 200mg/day INCREASED LEVELS DECREASED LEVELS HYPER PARATHYROIDISM TETANY RENAL STONES HYPERVITAMINOSIS D MULTIPLE MYELOMA
- 26. METHOD 5ml urine +5 drops of 1% acetic acid + 5ml of potassium oxalate. OBSERVATION : Trace amount of white precipitate INFERENCE : Calcium is precipitated as calcium oxalate in acid medium.
- 27. INORGANIC PHOSPHATE • Dailyexcretion : 1gm/ day INCREASED LEVELS DECREASED LEVELS RICKETS / OSTEOMALACIA DIARRHOEA HYPERPARATHYROIDISM NEPHRITIS ACIDOSIS PARATHYROID HYPOFUNCTION PREGNANCY
- 28. METHOD 5ml urine +few drops of conc HNO3 + a pinch of ammonium molybdate. Warm OBSERVATION Canary yellow precipitate. INFERENCE : Inorganic phosphate is precipitated as canary yellow ammonium phosphomolybdate.
- 29. AMMONIA • Daily excretion: 0.5-0.8gm/ day • Urinary ammonia is derived from glutamine and other amino acids by kidney. INCREASED LEVELS DECREASED LEVELS INGESTION OF ACID FORMING FOODS ALKALOSIS DIABETIC KETOACIDOSIS NEPHRITIS URINARY TRACT INFECTION
- 30. METHOD 5ml urine +2% sodium carbonate (till the red litmus turns blue). Boil. During boiling, hold a piece of moistened red litmus paper at the mouth of the test tube. OBSERVATION : Red litmus turns blue INFERENCE : Ammonia liberated turns red litmus blue.
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- 32. UREA • Daily excretion: 20-30 gm/day • Urea is formed in the liver. • End product of protein metabolism. INCREASED LEVELS DECREASED LEVELS FEVER LIVER DISEASE DIABETES MELLITUS METABOLIC / RESPIRATORY ACIDOSIS INCREASED INTAKE OF PROTEIN NEPHRITIS
- 33. SODIUM HYPOBROMITE TEST METHOD 2mlurine + Few drops of sodium hypobromite solution. OBSERVATION : Brisk effervescence INFERENCE : Hypobromite decomposes urea to give nitrogen gas.
- 34. SPECIFIC UREASE TEST METHOD Labeltwo test tubes as ‘TEST’ and ‘CONTROL’ Into TEST : 5ml urea solution + 2ml well mixed urease suspension. Into CONTROL : 5ml urea solution + 2ml urease suspension which is heated strongly. Incubate both TEST and CONTROL at room temperature for 15- 20mins. At the end, add 2 drops of phenolpthalein to both the tubes.
- 35. OBSERVATION In TEST :Pink Color In CONTROL : No pink color INFERENCE Urease converts urea into ammonia and carbonic acid. Under the Ph of the reaction condition, ammonia and carbonic acid are converted to ammonium carbonate, Ph goes above 9.5. Since the content are alkaline, phenolphthalein gives pink color.
- 36. URIC ACID • Dailyexcretion : 0.6-1gm/ day • End product of purine metabolism. • Increased levels of uric acid in urine is URICOSURIA INCREASED LEVELS DECREASED LEVELS LEUKEMIA PURINE FREE DIET CANCERS GOUT
- 37. PHOSPHOTUNGSTIC ACID REDUCTIONTEST METHOD 2ml urine + few drops of phosphotungstic acid + few drops of 20% sodium carbonate. OBSERVATION Blue color is formed INFERENCE Uric acid is a reducing agent in strong alkaline condition. It reduces colorless phosphotungstic acid to tungsten blue.
- 38. SCHIFF’S TEST METHOD Wet apiece of filter paper with a few drops of ammoniacal silver nitrate solution. Add 1 or 2 drops of uric acid solution on the same paper. OBSERVATION : Black color is formed INFERENCE : Uric acid reduces ammoniacal silver nitrate to metallic silver which is black in color.
- 39. CREATININE • Daily excretion: 2gm/day in males, 1gm/day in females. • Urinary creatinine is formed from muscle creatine. INCREASED LEVELS DECREASED LEVELS MUSCULAR DISORDERS RENAL FAILURE MYASTHENIA GRAVIS THYROTOXICOSIS STARVATION UNCONTROLLED DIABETES MELLITUS
- 40. METHOD Label 2 testtubes as ‘test’ and ‘control’. Into Test : 2ml urine + 2ml saturated picric acid+ few drops of 10% NaOH Into Control : 2ml water + 2ml saturated picric acid+ few drops of 10% NaOH OBSERVATION ‘TEST’ – Orange Color, ‘CONTROL’ – Yellow Color INFERENCE Creatinine reacts with alkaline picrate solution to form orange creatinine picrate.
- 41. ETHEREAL SULPHATE • 100mgof organic sulphate is excreted per day. TEST FOR DETECTING ETHEREAL SULPHATE METHOD 1ml urine + 2ml BaCl2 + 2ml concentrated HCl. Mix and filter. Divide the filtrate into two portions. Boil one and compare with the other (control)
- 42. OBSERVATION : Traceturbidity compared to that in control (red color may also be seen) INFERENCE : This test is done after removing the inorganic sulphate. Conc Hcl hydrolyses ethereal sulphate to inorganic sulphate which then gives precipitate with BaCl2.
- 43. UROBILINOGEN • Increased urobilinogenconcentration in urine is a sensitive index of liver dysfunction or hemolytic disorder. • Urobilinogen is present in excessive amount in prehepatic and hepatic jaundice. • Urobilinogen is absent in urine in post hepatic jaundice
- 44. METHOD 5ml of freshlyvoided urine + 1ml Ehrlich reagent, mix. Let stand for 5 mins. OBSERVATION Red color is seen when viewed through the mouth of the test tube. INFERENCE Urobilinogen reacts with p-dimethyl amino benzaldehyde of the reagent to give red color.
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