CHEMICAL EXAMINATION OF URINE

TEST PRINCIPLE REAGENT POSITIVE RESULT

ALBUMIN Heat & Acetic Acid Test Urinalysis protein is denatured by acid and heat so that it becomes less 10% Acetic Acid (+) cloudiness
Heller’s Test, Nitric Acid soluble & is precipitated & coagulated, respectively by acid & heat Conc. Nitric Acid
GLUCOSE Benedict’s Test When soluble copper sulfate is heated in strongly alkaline solution, the Benedict’s Reagent (+) orange to brick red precipitate
cupric ions are reduced to brick red insoluble cuprous ions (cuprous
oxide) by reducing agents such as glucose.
BENCE JONES Heat Precipitation Test The test involves heating the urine specimen with acetic acid and 5% acetic acid (+) precipitate dissolves upon
PROTEIN observing if the precipitate appears on heating, dissolves on further cooling to 56-60OC
heating and reappears on cooling.
ACETONE & Rothera’ Test Sodium nitroprusside is decomposed in alkaline solution. The resulting Ammonium sulfate (+) red-purple color
KETONE BODIES compounds are strong, oxidizing agents & in the presence and acetone Sodium nitroprusside
yield a rose- or purple- cpolred complex. The test is more sensitive for Ammonium hydroxide
diacetic acid than for acetone
Gunning’s Test Formation of yellow sediment of iodoform crystals results from the Strong ammonia (+) idoform crystals
reaction of Lugol’s solution Lugol’s solution
Gerthardt’s Test 10% chloride solution (+) Bordeaux red color
Hart’s Test 1 mL acetic acid (+) red ring
Hydrogen peroxide
LEVULOSE Seliwanoff’s Test Boiling urine with conc. HCl converts levulose to oxymethyl furfurol which Seliwanoff reagent (+) orange to red color
produces a red color when condensed with resorcinol
LACTOSE Rubner’s Test If lactose is present, the solution turns brick-red; red precipitate will Lead acetate (+) red precipitate & red
separate. The precipitate is the criterion for the test. Dextrose produces Strong ammonia water precipitate will separate
a red solution with yellow precipitate
BILE Gmelin’s Test Bile pigments are oxidized by acids forming a series of colored derivatives Nitric acid (+) play of colors: green on the
periphery, then red, and yellow
nearest the acid layer
BILIRUBIN Iodine-Smith Test 0.7% iodine & 95% ethyl (+) green ring
alcohol
UROBILINOGEN Ehrlich-Benzaldehyde Test The colorless urobilinogen is changed into a colored compound with Ehrlich-Benzaldehyde Cherry red color – increased
Ehrlich’s reagent Reagent amounts
Absence of cherry re od color –
decreased
UROBILIN Schlesinger Test Urobilinogen is oxidixzed to urobilin by the addition of Lugol’s Solution. Lugol’s Solution (+) fluorescent green color
The addition of zinc acetate leads to the production of a fluorescent green Zinc acetate solution
color
INIDICAN Obermayer’s Test Indoxyl potassium sulfate is hydrolyzed by strong metal and the resulting Obermayer’s reagent (+) indigo blue
indoxyl is oxidized tl indigo blue or to indigo red if the reaction is slow. Chloroform Normal: light blue

HEMOGLOBIN Guaiac Test 10% acetic acid, ether, 95% (+) blue color at the junction of
alcohol, guiac powder the guaiac and ether
Ammonium Sulfate Test Ammonium sulfate Clear supernatant – hemoglobin
Colored supernatant - myoglobin
MELANIN Thomalen Test Soidum nitroprusside is reduced to ferocyanide (Prussian blue) by 5% aqueous sodium (+) red color changes to blue
melanogen nitroprusside, 25% naOH, green
glacial acetic acid
CALCIUM Sulkowitch Test Calcium is precipated as calcium oxalate. This causes a milky turbidity Sulkowitch reagent (+) milky turbidity
CHLORIDE Fantus Test Silver nitrate reacts with chloride in the urine to precipitate silver chloride. Nitric acid (+) white precipitate
The nitric acid prevents the precipitation of silver phosphates Silver nitrate solution