been diluted greater than 20 X.
EXPERIMENT 2Tests for abnormal urinary components.
Detection of sugar in the urine.
The most common type of sugar that is found in the urine is glucose (in diabetes and in kidney failure) and lactose(sometimes during pregnancy or lactation). Medically, the presence of glucose or lactose in the urine is significant. Itis also important to differentiate between glucose and lactose. The presence of lactose is not indicative of a pathological condition but the presence of glucose at above a certain concentration indicates diabetes mellitus.
BENEDICT TEST FOR REDUCING SUGARS
1.Add 8 drops of urine to 5 ml of Benedict qualitative reagent in a test tube.2.Mix thoroughly and place the tube in a boiling water-bath for 5 min.3.The formation of orange-brown complex confirms the presence of reducing sugars.4.This reagent specifically shows positive result with reducing carbohydrates and does not react withaldehyde or any other forms of reducing agents.
FEARON TEST FOR GLUCOSE, LACTOSE AND MALTOSE
1.Boil 5 ml of glucose, lactose and maltose (in separate test tubes) with 0.5 ml of 5% methylamine-HCl.2.Then add 2 ml of hot, dilute NaOH.3.Cool to room temperature.4.Lactose and maltose produce red colour complex.5.Repeat the test using the urine sample.
Test for ketone bodies
The ketone bodies in the urine include acetoacetic acid, b-hydroxybutyric acid and also acetone. These compounds areexcreted in the urine when fats are metabolized excessively. Acetoacetic acid may easily undergo decarboxylation toform acetone.
ROTHERA TEST ( TO DETECT ACETON AND ACETOACETA TE)
1.Add ammonium sulphate crystals into 3 ml of urine and shake until it becomes saturated.2.Add a few drops of sodium nitropruside (5%, w/v).3.Gently add 2 ml of concentrated ammonia to the surface of the solution.4.The addition of ammonia causes the formation of a purple ring at the interface (between the urineand the ammonia) in the presence of ketone bodies.5.Shake the tube gently to ensure that the colour is homogeneously distributed. Note: the formation of the colour complex may take up to 5 - 10 minutes if the ketone concentration is very low.6.This test is sensitive up to 100 ppm (parts per million) acetone or 8 ppm acetoacetic acid in urine.
Test for proteins
In a healthy individual, very little protein (30 -200 mg/24 hr) is excreted in the urine. The amount of protein that isexcreted increases in pathological conditions such as heart disease, toxemia, pregnancy, kidney failure and liver 3