PRACTICUM II URINE CHEMICAL EXAMINATION 2.1 Glucose Practice : Urine Glucose Examination (Reduction Test) Objective : To determine the presence of glucose in the urine which issupporting examinations in confirming the diagnosis of the disease Diabetes mellitus(DM) Examination for the presence of glucose in the urine includes a filter examination. Determining the presence of glucose can be done in different ways. A non-specific method uses the properties of glucose as a reducing agent, in such tests there is a substance in the reagent that changes its properties and color when reduced by glucose. Among the many types of reagents that can be used to confirm the presence of reduction, cupric salts are the most widely used. Glucosuria can also be proven by a specific method that uses the enzyme glucose - oxidase to initiate a series of reactions and ends with a color change in the reagent used. 2.1.1 Benedict's Method Working principle: In an alkaline atmosphere in boiling reagents, glucose in urine can reduce cupric ions to cupro ions, so that the color changes from yellow to brick red. Tools and materials: 1. Tube Rack 2. Reaction Tubes 3. Wood clamp 4. Spirit lamp / water bath 5. Pipette 6. Benedict's solution Ways of working : 1. Put 5 ml of Benedict's reagent into a test tube. 2. Add 5 – 8 drops of urine to the tube. 3. Heat over a Bunsen flame for 2 minutes to boil or place in a boiling water bath for 5 minutes 4. The reaction results are observed and recorded Interpretation of Results: Negative (-) : Liquid remains blue in color Positive (+1) : Yellowish green in color Positive (+2) : Yellow in color Positive (+3) : Orange to Light Brown Positive (+4) : Brick Red 2.1.2 Method: Fehling Principle: The presence of glucose in urine will reduce cupric ions to cuprous When boiling, the urine will change color which is an indicator of the presence of glucose in the urine. Tools and materials : 1. Tube Rack + Test Tube 2. Wood Clamp 3. Spirit lamp 4. Pipette 5. Water bath 6. FEHLING A & FEHLING B solution (made in a 1:1 ratio) Ways of working : 1. Add the solvent mixture. Fehling (1:1) into a tube as much as 2 ml 2. Add 0.5 ml of urine and homogenize 3. Heat over a Bunsen flame for 2 minutes to boil or place in a boiling water bath for 5 minutes 4. The reaction results are observed and recorded Interpretation of results: Negative (-) : Liquid remains blue in color Positive (+1) : Yellowish green in color Positive (+2) : Yellow in color Positive (+3) : Orange to Light Brown Positive (+4) : Brick Red