ANALYSIS OF URINE

INTRODUCTION
Urine: Is an ultrafiltrate of plasma from which glucose, amino acids, water and other substances essential to body metabolism have been reabsorbed. Urine carries waste products and excess water out of the body.

URINE COLLECTION
There are basically four types of urine specimens: 1.First morning specimen 2.Random urine specimen 3.Fractional collection 4.Timed collection

 Time of analysis: - must analyzed within 1h at room temp. or within 8hr at 2oC- 8oC - If not assayed within these time limits, several changes will occur. sample should collected in a clean container. urine container must be sterile if the urine is to be cultured. For microscopic examination, the urine must be fresh

Urine Analysis:
Type of analysis: macroscopic analysis: 1) Physical examination 2) Chemical examination microscopic examination: urine sediment is
examined under microscope to identify the components of the urinary sediments.

Steps in basic urine analysis

Three steps analysis: First: physical characteristics of urine are noted and recorded. Second: series of chemical tests is run. Third: urine sediment is examined under microscope to identify the components of sediments.

PHYSICAL EXAMINATION
Physical examination involves: 1. Color 2. Transparency 3. Odour 4. Volume 5. pH 6. Specific gravity

1- Color: Many things affect urine color, including fluid balance, diet, medicines, and diseases. Color intensity of urine correlates to concentration. Darker color means more concentrated sample. Amber yellow Colorless Urochrome (derivative of due to reduced concentration. Pus, bacteria or
urobilin, produce from bilirubin degradation, is pigment found in normal urine).

Silver or milky appearance epithelial cells Reddish brown

Blood (Hemoglobin).

Yellow foam Bile or medications. Orange, green, blue or red medications. Vitamin B supplements can turn urine bright yellow.

TRANSPARENCY :
Urine is normally clear. Bacteria, blood, sperm, crystals, or mucus can make urine look cloudy. Is classified as clear or turbid. Degree of cloudiness depends on: pH and dissolved solids
Turbidity: may be due to gross bacteriuria, Smoky appearance: is seen in hematouria. Thread-like cloudiness: is seen in sample full of mucus.

ODOUR
Normal= aromatic due to the volatile fatty acids Ammonical bacterial action Fruity- ketonuria

- pH: pH measure acidicity or alkalinity (basic) of urine Normal urine pH: 4.5-8. Increased acidity in urine: due to diabetes or medications. Urine sample must be fresh (why?) (on standing urine become alkaline as a result of ammonia liberation due to urea decomposition). A urine pH of 4 is strongly acidic, 7 is neutral (neither acidic nor alkaline), and 9 is strongly alkaline.

Specific Gravity (SG): measures the amount of substances dissolved in urine. also indicates how well kidneys are able to adjust amount of water in urine. higher SG: more solid material is dissolved in urine When you drink a lot of fluid, your kidneys make urine with a high amount of water in it which has a low specific gravity. When you do not drink fluids, your kidneys make urine with a small amount of water in it which has a high specific gravity.

CHEMICAL EXAMINATION
Proteins Sugars Ketone bodies Bilirubin Bile salts Urobilinogen Blood

Tests for proteins

Test HEAT & ACETIC ACID TEST Principle-proteins are denatured & coagulated on heating to give white cloud precipitate. Method-take 2/3 of test tube with urine, heat only the upper part keeping lower part as control. Presence of phosphates, carbonates, proteins gives a white cloud formation. Add acetic acid 1-2 drops, if the cloud persists it indicates it is protein(acetic acid dissolves the carbonates/phosphates)

TEST FOR SUGAR

Test-BENEDICTS TEST(semiquantitative) Principle-benedicts reagent contains cuso4.In the presence of reducing sugars cupric ions are converted to cuprous oxide which is hastened by heating, to give the color. Method- take 5ml of benedicts reagent in a test tube, add 8drops of urine. Boil the mixture.

SUGAR DETERMINATION (Benedict test):blue-sugar absent; green-0.5% sugar yellow-1% sugar; orange-1.5% sugar; brick red-2 % or more sugar.

KETONE BODIES
3 types Acetone Acetoacetic acid -hydroxy butyric acid They are products of fat metabolism

Rotheras test
Principle-acetone & acetoacetic acid react with sodium nitroprusside in the presence of alkali to produce purple colour. Method- take 5ml of urine in a test tube & saturate it with ammonium sulphate. Then add one crystal of sodium nitroprusside. Then gently add 0.5ml of liquor ammonia along the sides of the test tube. Change in colour indicates + test

BLOOD IN URINE
Test- BENZIDINE TEST Principle-The peroxidase activity of hemoglobin decomposes hydrogen peroxide releasing nascent oxygen which in turn oxidizes benzidine to give blue color. Method- mix 2ml of benzidine solution with 2ml of hydrogen peroxide in a test tube. Take 2ml of urine & add 2ml of above mixture. A blue color indicates + reaction.

Urine strip:
Strip is filter paper or plastic which has chemical substance (reagent) coated on it on different pads. It gives color when react with substance in urine. The produced color is compared with chart color visually or mechanically assessed.

Microscopic examination
Microscopic urinalysis is done simply pouring the urine sample into a test tube and centrifuging it (spinning it down in a machine) for a few minutes. The top liquid part (the supernatant) is discarded. The solid part left in the bottom of the test tube (the urine sediment) is mixed with the remaining drop of urine in the test tube and one drop is analyzed under a microscope

Crystals in urine
Crystals in acidic urine Uric acid Calcium oxalate Cystine Leucine Crystals in alkaline urine Ammonium magnesium phosphates(triple phosphate crystals) Calcium carbonate

CRYSTALS

CASTS
Urinary casts are cylindrical aggregations of particles that form in the distal nephron, dislodge, and pass into the urine. In urinalysis they indicate kidney disease. They form via precipitation of Tamm-Horsfall mucoprotein which is secreted by renal tubule cells

Types of casts
Acellular casts Hyaline casts Granular casts Waxy casts Fatty casts Pigment casts Crystal casts Cellular casts Red cell casts White cell casts Epithelial cell cast

WAXY CAST

FATTY CAST

REFERENCE
1)Urinalysis - MayoClinic.com 2)http://www.mdconsult.com/das/book/body/208746 819-6/0/1445/0.html. 3)ibrary.med.utah.edu/WebPath/TUTORIAL/URINE/UR INE.html 4) en.wikipedia.org/wiki/Urinalysis 5) https://ahdc.vet.cornell.edu/clinpath/modules/uased/ua-intro.htm 6) www.bloomberg.com/.../iphone-urinalysis-drawsfirst-fda-inquiry-of-me.

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