Urine Examination

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Urine examination
Introduction
Urine is the waste product in ourbody.The kidneys formurine which passes through the ureters to the bladderforstorage
prior to excretion. Waste productsofprotein metabolismare excreted,electrolyte balance is maintained and the pH (acid-
base balance) is maintained by the excretion of hydrogen ions. Nephron is the functional unit of kidney.
Examination of urine is a fundamentalinvestigation in patients in whomkidney disorders orinfections ofthe urinary tract
are suspected.There are alsomany patientswhoexhibit no clinicalsymptoms,butin whompreviously unrecognized urinary
tract infections can be diagnosed by urine examination.
There are three processes involved in the formation of urine:
1. Simple filtration
2. Selective re-absorptions
3. Secretion.
Composition of urine:
Water …………………96%
Urea ……………………2%
Ammonia
Calcium
Chlorides
Creatinine
Magnesium ……….. 2%
Oxalates
Phosphates
Potassium
Sodium
Sulphates
Uric acid
Few Terminologies
1 Pyuria:
Pus cells present in urine is called pyuria.
Normally pus cells are absent in urine but some time may be seen in 1-3/HPF in male and 1-5/HPF in female which in
normal but if there is >5/HPF indicates abnormal.
2. Bacteriuria:
Bacteria present in urine are called bacteriuria.In normal urine, there is absent bacteria but long time incubationthere
may be bacteria because ofmultiplication ofbacteria.(Urine is good media forbacteria). So,urine examination should
be examined within 1-2 hour of collection. If not possible urine should be preserved.
3. Haematuria
Blood present in urine is called haematuria.
4. Nephritis:
Inflamation of nephron called nephritis.
5. Cystitis:
Inflammation of urinary bladder.
6. Polyuria:

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Excessive output of urine. (>2000 ml).
Conditions- Diabetes Mellitus, Diabetes insipidus, Recovery fromacute renal failure (ARF), Diuretic therapy
7. Oligouria:
Less than 500 ml of urine per day in adults,
Severe oliguria (<100 ml of urine per day in adults)
Conditions- ARF, Vomiting, Fever, Burns.
8. Nocturia:
Excessive urination at night is called Nocturia.
9. Dysuria:
Painful urination is called dysuria.
10. Anuria:
Absence of urination or < 50 ml per 12 hours.
1. Renal threshold: The maximum capacity of kidney to absorbe glucose i.e 160-180mg/dL is called renal threshold.
Glucose appears in the urine when the blood glucose level is more than 10 mmol/l (180 mg%).
12.Renal glycosuria:It is a conditionin which the renalthresholdforglucoseis decreases.Glomerularfiltration rate (GFR)
will be normalbut defect on the renalabsorption.It is unrelated todiabetes mellitus.It does not showany symptoms of
diabetes.
Volume of urine: 0.7 to 2 litre/ day (24 hrs).
Collection of urine specimens
Containers forthe collection ofurine should be wide-mouthed,clean and dry.Labelpatient name,lab no,age/sex,date etc.
If the urine specimen has to be transported for any length of time it should contain an appropriate preservative to prevent
bacterial overgrowth or hatching of viable ova.
Types of urine specimen
1. Early morning urine specimen
Early morning urine provides the most concentrated sample. It is better for urine pregnancy test.
2. Random urine specimen
A randomurine sample, taken at any time of the day, will enable the laboratory to screen for substances which are
indicators of kidney infection.
3. 24-Hour urine specimen
The 24-hour urine specimen is collected in a clean gallon with a stopper.On the first morning the patient getsup and
urinates; this urine is not collected.Allthe urine passed during the rest ofthe day and night is collected in the bottle.
The next morning the patient getsup and collects thefirst urine ofthe morning in the bottle (8 amto 8am). The bottle
should then betaken immediately to the laboratory.Measure the volume ofurine with a measuring cylinderand record
it.
4. Midstream urine specimen
While passingurine,the patient placesan opencontainerin the streamofurine and collects about 20to 30 ml of urine.
The container should be covered immediately. This sample is suitable of urine culture.
5. Catheter urine sample
Collection of urine using a catheter must be carried out by a qualified physician or nurse. The procedure is used for
certain bacteriological tests, mainly in women. Usually, however, a specimen collected in the normal way following
thorough cleansing is acceptable for this purpose.

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6. Suprapubic aspiration
This method is suitable for infants. Sample is collected by physician or experience nurse or other medical person.
Preservation of urine specimens
Urine passed at a clinic and examined immediately, does not require preservation.
Preservative methods for urine sample.
1. Refrigerator method
2. Chemical method
1. Refrigerator method:
In refrigerator 2-8°C is better.
2. Chemical methods
Mainly acids are usedas a chemical preservative.
i. Concentrate HCl: 10-30 ml HCL is used to 24hrs urine sample. This is suitable for calcium, phosphate, urea,
creatinine urine test.
ii. Toluene: 5-10 ml is used to 24hrs urine sample. This is suitable for protein, uric acid etc.
iii. Chloroform: Same as toluene.
iv. Acetic acid: 10 ml is used to 24hrs urine sample. This is suitable for vitamin test.
v. Boric acid: This is most common preservative used in lab. 1-2 grams boric acid is used in 120 ml of urine. It is
also used in urine culture. 10 gmfor 24hrs urine.
vi. Formalin: 1-2 drops formalin is used in 30ml of urine.
vii. Thymol: 0.1gm per 100ml of urine sample.
Urine Routine Examination
Collection of urine specimens
Containers for the collection of urine should be wide-mouthed, clean and dry with screw capped. Label patient name, lab
no, age/sex, date etc. If the urine specimen has to be transported for any length of time it should contain an appropriate
preservative to prevent bacterial overgrowth.
Urine routine examination:
1. Physical examination (Macroscopic examination)
2. Chemical examination
3. Microscopic examination test
1. Physical examination:
Physical examination is done by naked eye.
a. Colour
b. Transparency
c. pH

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d. Specific gravity (Sp. G)
Colour:
Urine is normally clear straw-yellow or light yellow in colour. More concentrated urine may appear dark yellow.
The colour of urine is light yellow or straw yellow because present of urobilin.
The presence of blood cells or excess salts may make the urine appear cloudy.
Pigments frombile substances may make the urine appear deep yellow or brown.
Appearance Possible Cause
Cloudy ● Bacterial urinary infection
Red and cloudy
(Due to red cells)
● Urinary schistosomiasis
● Bacterial infection
Brown and cloudy
(Due to haemoglobin)
● Malaria haemoglobinuria
● Other conditions that cause
intravascular haemolysis.
Yellow-brown, orgreen-brown
(Due to bilirubin)
● Acute viral hepatitis
● Obstructive jaundice
Yellow-orange
Due to urobilin i.e. oxidized
urobilinogen
● Haemolysis
● Hepatocellular jaundice
Milky-white ● Filariasis, Pus
Colourless Very dilute urine, polyuria
Transparency
Normally fresh voided urine is clear. If urine is not clear we report as turbid. The following condition urine may turbid,
1. UTI
2. Filariasis
3. High protein present in urine.
Specific gravity
Measured by urinometer - implies the capacity of kidney to concentrate urine.
Normal value: 1.015-1.030. Depends upon - State of hydration & solute load
Increased specific gravity is called hypersthenuria.
Decreased specific gravity is called hyposthenuria.
Values more than 1.030 indicates
 Severe dehydration
 DM (diabetes mellitus)
 Adrenal insufficiency
Values less than 1.015 indicates
 Increase water intake
 Diabetes insipidus
 Chronic Nephritis

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Specific gravity of urine can be measuredwith
1. Urinometer 2. Refractometer
3. Dip Sticks 4. Osmometer
5. Specific gravity bottle 6. Specific gravity beads
Fig: Urinometer
Fig: Urinometers
pH
Normal freshly passedurine is slightly acid (around6.0).In certain conditions,the pHofthe urine may increaseordecrease.
Normal range is 4.5 to 8.5.
Average is 6.0
Determination of urine pH
1. By using pH meter
2. By using pH strip
Increasedcondition: After heavy meals, Proteus infection, Dietary products.
Decreasedcondition: Afterheavy exercise,Metabolic Acidosis,Chronic respiratory acidosis,vegetarian diet,high protein
intake.

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2. Chemical examination:
a. Urine sugar test
b. Urine protein test or urine albumin test
URINE SUGAR TEST:
Normally sugar is absent in urine. If glucose is present in urine is called glycosuria. Glycosuria is condition of diabetes
mellitus. We can performurine sugar by following methods.
i. Benedict's is method
ii. Urine strip method
iii. Fehling test
Urine sugar test by Benedict's method:
Benedict’s test is used for testing reducing sugars. This test is also called ad reducing sugar test. A reducing sugar is a
carbohydratepossessing eithera free aldehydeorfree ketone functionalgroupas partofits molecularstructure.This includes
all monosaccharides(eg.glucose,fructose,galactose)andmany disaccharides,including lactoseandmaltosebut not sucrose
(sucrose is non-reducing sugar).
It can be used as a semi-quantitative test.
Benedict’s test is most commonly used to test forthe presence ofglucose in urine. Glucose found to be present in urine is
an indication ofDiabetes mellitus. Glucose is a reducing substance.It reduces the blue coppersulfate in benedict solution
to red copper oxide, which is insoluble.
Principle:
Reducing sugars under alkaline and heat conditions, copper sulphate (Cu2+
) reduced to cuprous oxide (Cu2O) and gives
green, yellow, orange or brick red precipitate.
Cupric ions (Cu2+
) + Urine glucose
𝐴𝑙𝑘𝑎𝑙𝑖𝑛𝑒 𝑐𝑜𝑛𝑑𝑖𝑡𝑖𝑜𝑛
𝐻𝑒𝑎𝑡
> Cuprous oxide (Cu2O)
(Blue colour)
Precipitate to green, yellow, orange or brick red.
Materials and reagents
1. Test-tubes
2. Wooden test-tube holder
3. Test-tube rack
4. Beaker
5. Bunsen burner or spirit lamp
6. Dropper pipette
7. Graduated pipette, 5ml
8. Benedict solution.

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Procedure:
1. Pipette 5 ml of Benedict solution into a test-tube.
2. Add 8 drops (0.5ml) of urine and mixwell.
3. Boil overa Bunsen burnerorspirit lamp for 2 minutes orplace the test-tube in a beakerat boiling waterfor5 minutes.
4. Place the test-tube in the test-tube rack and allow to cool to roomtemperature.
5. Examine the colour change of the solution and any precipitate, report the result.
Reporting
1. No colour change (blue) Nil
2. Slightly colour change  Trace
3. Green colour  + (0.5%)
4. Yellow colour  ++ (0.5-1.5%)
5. Orange colour  +++ (1.5 - 2%)
6. Red colour (brick colour)  ++++ (>2%)
Precautions:
1. Used clean and dry test tube.
2. Benedict’s solution should be clean and dry.
3. Test tube mouth should be away fromface.
Composition of Benedict's solution:
1. Solution A:
i. Copper sulphate  17.3 gram
ii. Distilled water  300 ml
2. Solution B:
i. Sodiumcarbonate  100 gram
ii. Sodiumcitrate  175 gram
iii. Distilled water  700 ml
Mix Solution A and B then pour on bottle and label it and store at roomtemperature.
 Copper sulphate provides cupric ion (Cu++ ion) in solution.
 Sodium citrate prevents precipitation of cupric ion as cuprous hydroxide by forming a loose cupric sodium citrate
complex.

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 Sodiumcarbonate provides the alkaline pH to the solution
False positive results
1. Chloral hydrate
2. Isoniazid
3. Aminosalicylic acid
4. Nalidixic acid
5. Nitrofurantoin
6. Vitamin C.
7. Glucuronides.
8. Patient under dextrose infusion.
9. Thymol
10. Chloroform.
ii. Urine sugar detection by stripmethod
Principle:
Glucose is oxidized to gluconolactone by the enzyme glucose oxidase (specific for glucose) with the release of hydrogen
peroxide.A chromogen isthen oxidized by the hydrogenperoxide and a peroxidase enzyme is usedto convert the chromogen
from a reduced colourless state to a coloured oxidized state.
Advantages of strip method:
1. Quick
2. Cheap
3. Easy to perform
4. More specific
5. More sensitive
URINE PROTEIN TEST
Normally healthy people’surine contains only a smallamount ofurinary proteins (<150 mg perday)but not detectprotein
in urine by quantitative method.Normally we cannot detectprotein in urine.If protein detect in urine is called proteinuria.
Albumin is the commonest protein excreted in urine due to its relatively small size (molecular weight 66,000) and high
concentrationin plasma.Proteinuria is an importantpathologicalfinding with severalimportant causes,mostly renalorigin.
Methods of urine protein detection:
i. Heat and acetic acid method
ii. Sulphosalicylic acid method
iii. Urine strip method
Heat and acetic acid method:
Principle:
Heat causesthe proteinsin urine to coagulate.Proteins are more
susceptible to precipitation whenthey are at the pHoftheir isoelectric point,
which is usually low.
Requirements
1. Test tubes and test tube holder

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2. Spirit lamp
3. Dropper
4. Timer
5. Reagent  5% acetic acid
6. Sample  urine
Procedure:
1. Take 3-5 ml of urine in a test tube.
2. Heat the upper portion of the urine.
3. Add 2-3 drops of 5% acetic acid.
4. Observe the white coagulumor turbid against a dark background.
Result:
1. No change (Clear)  Nil
2. Slightly cloudy  Trace
3. Cloudy  + (Average 30 mg/dl)
4. Cloudy with small precipitate  ++ (Average 100 mg/dl)
5. Cloudy with moderate precipitate  +++ (Average 300 mg/dl)
6. Cloudy with large precipitate  ++++ (Average 1000 mg/dl)
ii. Urine protein test by Sulphosalicylic acidmethod
This method is sensitive and commonly uses method for urine protein test.
Principle
Protein in the urine is precipitatedbysulphosalicylic acid.The degree ofturbidity is relatedto the quantityofprotein present
in urine.
Requirements
1. Test tube with test rack
2. Test tube holder
3. Timer
4. Pipette
5. Sample urine
6. Reagent 3% Sulphosalicylic acid
Procedure
1. Take 3ml of urine taken in a test tube,
2. Add 3ml of 3% Sulphosalicylic acid and mixwell.
3. Observe the result against a dark background.
Note: Control (C) also required.
Result
1. No change (Clear)  Nil
2. Slightly cloudy  Trace
3. Cloudy  +
4. Cloudy with small precipitate  ++

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5. Cloudy with moderate precipitate  +++
6. Cloudy with large precipitate  ++++
Protein test by stripmethod:
Urine protein strip tests detect mainly albumin. The test area is impregnated with the indicator tetrabromophenol blue. In
the presence of protein there is a change in the colour of the indicator fromlight yellow to green-blue depending on the
amount of protein present in urine.
Clinical significance
Proteinuria is seen in following conditions.
1. Urinary tract infection (UTI)
2. Glomerulonephritis
3. Diabetic nephropathy
4. Pyelonephritis
5. Nephrotic syndrome
6. Sever illness
7. Last stage of pregnancy
8. Fever
9. High protein diet
10. Infection
11. Toxiemia of pregnancy
12. Systemic Lupus Erythematosus (SLE)
13. Exposure to severe cold
14. Multiple myeloma
Microscopic Examination
Microscopic examination of urine is done form the deposit of urine.
1. Take clean and dry test tube and label it.
2. Pour urine around 3-5 ml in test tube.
3. Centrifuge at 3000 rpmfor 3-5 minutes.
4. Then discard supernatant part and pour deposit part in center of slide.
5. Cover with cover slip then observe on microscope at 10X the 40x.
Microscopic findings of urine:
i. RBCs
ii. Pus cells
iii. Epithelial cells
iv. Yeast cells
v. Bacteria
vi. Spermcells
vii. Casts

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viii. Crystal
ix. Parasites
X. Mucous threads
1. RBCs:
Normally RBC is not presentin urine.More than one RBC perhigh powerfield is abnormal.The presence ofRBC in
urine is called haematuria.
The red blood cells swell up and broken down in diluted urine. The red cells may show crenated margins.
Clinical Significance: Red blood cells are found in
a) Pyelonephritis b) Renal stones c) Cystitis d) Polycythemia vera
Normal RBC
Crenated RBC
Swelling RBC
Figure of RBCs
2. Pus cells:
Pus cells present in urine is called pyuria.Normally pus cells are absent in urine butsome time may be seen in 1-3/HPF
in male and 1-5/HPF in female which in normal but if there is >5/HPF indicates abnormal.
Large number of pus cells indicates bacterial infection of urinary tract.
Figure: Pus cells
3. Epithelial cells:
These cells have a single rounded nucleus. Squamous epithelial cells present in urine in moderate numbers have no
pathological importance.

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Clinical Significance: Presence of all other epithelial cells indicates pathological condition.
Renal tubular epithelial cells: Unstained cells have almost the same size as that of a neutrophil but contain a large
round nucleus. Oval fat bodies are the cells containing fat globules. The nucleus, is not visible.
Bladder epithelial cells: Unstained cells are larger than renal tubular cells having a round nucleus and vary in size
depending on depth of origin.
Squamous epithelial cells: These are unstained large flattened cells with abundant cytoplasmand a small round
nucleus. The cell may be folded or rolled.
Figure: Epithelial cells
4. Bacteria:
Bacteria are not normally present in urine. However, unless specimens are collected under sterile conditions,a few
bacteria are usually present as a result of vaginal, urethral, external genitalia, or collection-container contamination.
These contaminant bacteria multiply rapidly in specimens that remain at room temperature for extended periods, but
are of no clinical significance. They may produce a positive nitrite test result and also frequently result in a pH
above 8, indicating an unacceptable specimen.
Bacteria may be present in the form of cocci (spherical) or bacilli (rods). Owing to their small size, they must be
observed and reported using high-power magnification.
They are reported as few, moderate, or many per high-power field. To be considered significant for UTI, bacteria
should be accompanied by WBCs.
Figure: Bacteria

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5. Yeast cells:
Yeast cells appear in the urine as small, refractile oval structures that may or may not contain a bud. In severe infections,
they may appear as branched, mycelial forms. Yeast cells are reported as rare, few, moderate, or many per hpf.
Differentiation betweenyeastcells andRBCs can sometimes bedifficult.Carefulobservationforbuddingyeast cells should
be helpful.
In normal urine yeast cells are absent whereas,in case ofdiabetic,immunocompromised patients andwomen with vaginal
moniliasis may present yeast cells.
Fig: Yeast cells
6. Sperm cells
Spermatozoa are easily identified in the urine sediment by theiroval,slightly tapered heads and long tail.Spermatozoa are
occasionally found in the urine in male. In some cases of both men and women contain spermcells in urine in sexual
intercourse,masturbation,ornocturnalemission.They are rarely of clinical significance except in cases ofmale infertility
or retrograde ejaculation in which spermis expelled into the bladder instead of the urethra.
Figure: Sperm cells
7. Casts:
Urinary casts are formed in the lumen of the tubules of the nephrons and are originate from Tamm-Horsfall- protein
(molecular weight 18,000) secreted by renal tubules.
Casts have paralleledges,round or broken.They may be curved orstraight,short orlong.A significant numberofurinary
casts usually indicates the presence of renal disease.
Urinary casts can be divided into two main group: Acellular and Cellular Casts.
Acellular Casts Cellular Casts

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Hyaline Casts Red Blood Cell Casts
Granular Casts White Blood Cell Casts
Waxy Casts Bacterial Casts
Fatty Casts Epithelial Cell Casts
Casts Composition AssociatedConditions
Hyaline casts
Solidified Tamm-Horsfall
mucoprotein
 Normal individuals
 Dehydration
 Heavy exercise
Granular casts
Various celltypes (Degenerationof
cellular casts, Aggregates of
plasma proteins or
immunoglobulin light chains)
 After sternous exercise
 Chronic renal diseases
 Acute tubular necrosis
Waxy casts (renal failure
casts)
Various cell types (Final stage of
degeneration of cellular cast)
 Severe chronic renal disease
 renal amyloidosis
Fatty casts
Lipid droplets within the protein
matrix of the cast
 Tubular degeneration
 Nephrotic syndrome
 Hypothyroidism
RBC Casts Red Blood Cells
 Pyelonephritis
 Glomerulonephritis
 Acute interstitial nephritis
 Lupus nephritis
WBC Casts White Blood Cells  Glomerulonephritis
Epithelial Cell Casts Renal Tubular Epithelial Cells
 Renal tubular necrosis
 Viral Diseases
 Kidney transplant rejection

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Figure: Casts
8. Crystals:
Crystals are not usually found in freshly voided urine but appearafterthe urine stands fora while. Crystals are considered
"normal" if they are fromsolutes that are typically found in the urine. Some examples of crystals that can be found in
the urine of healthy individuals includes:amorphous urates,crystalline uric acid, calcium oxalates,amorphous phosphates
and calciumcarbonate.
If the crystals are from solutes that are not normally in the urine,they are considered “abnormal." Abnormalcrystals may
indicate an abnormal metabolic process. Some of these include: cystine, tyrosine and leucine. When crystals
form as urine is being made in the kidney, they may group together to form kidney "stones" or calculi. These stones can
become lodged in the kidney itself or in the ureters, tubes that pass the urine fromkidney to the bladder, causing
extreme pain. Medications,drugs,and x-ray dyes can also crystallize in urine. Associated with renalcalculi, commonly is
oxalate in acidic urine and phosphate in alkaline urine.

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Different types of crystals,
Normal Crystals Abnormal Crystals
1. Uric acid Crystals 1. Bilirubin Crystals
2. Calcium Oxalate Crystals 2. Cholesterol Crystals
3. Hippuric Crystals 3. Cysteine Crystals
4. Calcium Phosphate Crystals 4. Leucine Crystals
5. Triple Phosphate Crystals 5. Tyrosine Crystals
6. Calcium Carbonate Crystals 6. Sulfa Crystals
7. Ammonium Biurate Crystals 7. Indinavir Crystals

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18. 18

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Normal crystal

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Abnormal crystal

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9. Mucous threads
These are long thin waxy threadsofribbon like structures.These may be present in normalurine but found in inflammation
or irritation of the urinary tract.
10. Urine parasites:
a. Trophozoite of Trichomonas vaginalis:
b. Ova of Schistosoma haematobium
Schistosoma haematobium is established by demonstration of eggs in urine. The specimen should be immediately
centrifuged and the sediment examined by wet mount.
Trichomonasvaginalis motile trophozoitesmay also be foundin the urine,especially in infected male patients.To lookfor
the presence of trophozoites, the urine specimen should be centrifuged and, the sediment mixed with a drop examined by
wet mount. Temporary stains, such as methylene blue or malachite green, are also helpful.
Fig: Trophozoite of Trichomonas vaginalis Fig: Ova of Schistosoma haematobium

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Fig: Mkicroscopic findings in Urine

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Special Test
1. Ketone Bodies Test or (Acetone Test)
2. Chyluria Test (Urine chyle Test)
3. Urine Bile Salt Test
4. Urine Bile Pigment Test
5. Urobilinogen Test
1. Ketone bodies test or Acetone test in urine
The present of ketone body in the urine is called ketonuria. Ketonuria seen in mainly uncontrolled diabetic mellitus (type
1), Starvation, Severe vomiting, Glycogen storage disorders, High fat diet.
Acetoacetate, beta-hydroxybutyrate and acetone are collectively referred to as ketone bodies or ketones.
Formation of ketones:
The metabolismof glucose normally provides the body with its energy requirements.If, however,the intake of glucoseis
insufficient as in starvation,orglucosemetabolismis defectivedueto a lackofinsulin as occurs in untreatedoruncontrolled
diabetes,thebody obtainsits energyby breaking down fats.It is this increase in fat metabolismwhich leads to a buildup of
ketones in the body. An accumulation of ketones in the body is called ketosis.
Ketones are toxic to the brain and if present in sufficiently high concentrationin the blood they can contribute to the coma
found in diabetic ketoacidosis.Metabolic acidosis is causedbya variety ofnon-volatile acids accumulatingin the plasma of
patients with diabetic coma.Urine ketone tests detect acetoacetate and acetone. Beta hydroxybutyrate is not detected.
Methods of Ketone bodies test
a. Rothera's test
b. Gerhardt's test
c. Strip test
a) Rothera’s Test
Principle
Ketone bodies react with sodiumnitroprusside at alkaline condition to give purple colour complex.
Requirements:
i. Test tubes
ii. Dropper
iii. Marker
iv. Sample - urine
v. Reagent - Rothera's powder
Rothera's powder
Composition
1. Ammoniumsulphate crystals……….100gm
2. Sodiumnitroprusside crystal.………..3gm
3. Sodiumcarbonate.………………….50 gm

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Procedure
1. Take pea sized Rothera’s powder and pour in test tube.
2. Add few drops of urine in same tube.
3. Observe the result.
Result
No colour change = Negative
Purple colour = Positive
Note: Excess of sodium nitroprusside in reagent may give false positive result.
Ketone striptests
Severalketone strip testsare available commercially.The reagentstrip is impregnatedwith sodiumnitroprusside buffer.
Interpretations
1. Untreated diabetes mellitus
2. Starvation
3. Prolonged vomiting
4. Glycogen storage disease.
Ketone testing is most often done if have type 1 diabetes and:
 Blood sugar is higher than 240mg /dL
 Have an illness such as pneumonia, heart attack, or stroke
 Nausea or vomiting occur in pregnant

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2. Urine Chyle test
Introduction:
Present ofchyle in urine is called chyluria.It is complicationof bancroftian filariasis.It occurs whentheurogenitallymphatic
vessels whichare linked to thosethat transportchyle fromthe intestine become blocked andrupture.Milky white appearance
seen when chyle presents in urine.
Principle:
When urine is mixed with organic solvents like ether, chloroform, if urine contain chyle then urine become clear.
Requirements
a. Test tubes
b. Pipette
c. Sample - urine
d. Reagent - organic solvent e.g: chloroform, benzene or diethyl-ether
Procedure:
1. Take one clean and dry test tube and label it.
2. Add 5 ml urine in the test tube.
3. Add 1 to 2 ml reagent.
4. Observe the result.
Result:
If white part disappears or urine is clear: Positive
If no change: Negative
Clinical significance:
This test is used to diagnosis for filariasis.
Bile Salts Test In Urine
Bile salts are normally not excretedin urine but in obstructive jaundice,theyappearin urine.These are sodiumandpotassium
salt of taurocholic acid, glycocholic acid which assists in the emulsion and absorption of dietary lipid by reduce surface
tension.Bile salts are formed in the liverfrom cholesterol.Ifthere is obstructionin bile flow orleakage ofbile in blood then
this salt will appear in urine.
Method:
Hay’s test
Principle
Bile salts reduce the surface tension of urine, which causes sulfur powder to sink to the bottom.
Requirements

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1. Test tubes
2. Pipette
3. Sample - urine
4. Reagent - Sulphur powder
Procedure
1. Take 2 ml of urine in a test tube.
2. Sprinkle or spray sulfur powder in upper part of urine sample. Do not shake or mix.
3. If the sulfur powder sinks to the bottom, it indicates the presence of bile salts. Run a control using distilled water
instead of urine.
Result
Sulphur powder sink = Positive
Sulphur powder float = Negative
Interpretation
Positive in obstructive jaundice
Bile Pigment test (Urine bilirubin test)
Normal urine does not contain bile pigment.In certain liver disease (jaundice),anemia and infectious disease bile pigments
may pass into blood streamand excreted in urine.
Urine bile pigment test
1. Fouchet's test
2. Foamtest
3. Lugol’s Iodine method
4. Urine strip test
1. Fouchet's test:
Principle
Wet bariumsulphate hasthe capacity to absorbyellowbilirubin. Fouchet’s reagent oxidizes the absorbedbilirubin to green
color biliverdin & blue color bilicyanin.
Requirements:
1. Filter paper
2. Test tubes
3. Funnel
4. Sample (urine)
5. Reagent (i) Barium chloride
(ii) Fouchet's reagent
Composition of fouchet's reagent:
1. Trichloroacetic acid = 25 gram
2. 10% ferric chloride = 10 ml
3. Distilled water = 90 ml
Specimen:

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Freshly passed urine is required. It should be protected from daylight and fluorescent light because bilirubin is rapidly
oxidized by ultraviolet light to biliverdin which is not detected by the reagents used in the tube or strip tests.
Procedure
1. Take 10 ml of urine in a test tube.
2. Add 5ml of bariumchloride solution followed by pinch of magnesiumsulphate powder.
3. Mix well and keep the solution for 5 minutes.
4. Filter the solution and fold the filter paper and dry it.
5. Add few drops of fouchet’s reagent to the dry unfiltrate precipitate on the filter paper.
6. Observe the colour.
Result
1. Blue green colour = Positive
2. No colour = Negative
Interpretation
Bile pigments are found in urine along with bile salts in severe hepatitis & obstructive jaundice.
Urobilinogen test in Urine
Urobilinogen is normally presentin urine in low concentrations (<4mg/ 24 hrs).It is formed in the intestine frombilirubin,
and a portion of it is absorbed back into the bloodstream.
Increased amount ofurobilinogen in urine indicate liverdisorders in which liver function is impaired. Urobilinogen is high
in haemolytic jaundice but normal in obstructive jaundice.
Method:
1. Ehrlich’s test
2. Strip test
Specimen:
Freshly passed urine is requiredbecause urobilinogenwhich is colourless is rapidly oxidized to the orange pigmenturobilin
which is not detected by Ehrlich’s test or by urobilinogen strip tests.
Ehrlich’s test
Principle:
Urobilinogen reacts with p-dimethylaminobenzaldehyde to form a red condensation product. The intensity of colour
produced corresponds to the concentration of urobilinogen present.
Requirements:
1. Test tubes with test tube rack
2. Pipette
3. Sample = urine
4. Reagent; Ehrlich's reagent
Composition of Ehrlich's reagent:
1. P- dimethylaminobenzaldehyde: 2 gm
2. HCl = 20 ml

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3. Distilled water: 80 ml
Procedure:
Step1
1. Take 5 ml urine in test tube and add 5 ml Ehrlich’s reagent and mixwell.
2. Wait for 10 minutes.
Pink or red: Positive may be present and process further for confirmation.
Negative: No further confirmation
Step2
3. Mix 5 ml above mixture with 5 ml of saturated sodiumacetate solution.
Colour get intensity of excess of urobilinogen is present and process is further or confirmation.
Step3
4. Mix 5 ml above mixture with 5 ml chloroform, mix thoroughly and allow to separate layer.
Observation:
Lower chloroform= Pink or red = Positive for urobilinogen.
Control
Sometimes colouris faint or doubtfulin such case put a negative control(urine 6N HCL and process in similar as for
urobilinogen).
Interpretation
Increases
1. Hemolytic disease
2. Hepatocellulardamage orhepatic congestion,resultingin less ofthe absorbedurobilinogen being excretedby the liver.
3. Cirrhosis of the liver.