University:
Saint Louis University
Course:
PATH 0100 | Introduction to Pathology
Academic year:

2024
Views:

459

Pages:

3
Author:

brian 11kizz

V. ALBUMIN / PROTEIN 1. Protein with Sulfosalicyl Acid The purpose of protein with Sulfosalicyl Acid is to determine the presence of protein in the urine. Principle : A precipitate reaction is formed between sulfosalysyl acid and protein in the urine which appears white turbidity like a cloud. Material : Fresh urine Tools and Reagents Tool : a. Test tube b. Pastor pipette c. Math pipette 10 ml Reagents : Sulfosalysyl acid 20 % : Ways of working 1. Pipette 5 ml of urine and put it in a test tube located in the tube rack 2. Place 2-3 drops of 20% Sulfosaysil acid reagent in the urine tube. 3. Shake the tube then read and record the results. Interpretation of Results Negative (-) : If the urine remains clear Positive (+) : If turbidity occurs 2. Protein with Acetic Acid Principle : Protein withwhich appears to be cloudy. Acetic acid is administered to reach or approach the isoelectric point of the protein. Further heating causes denaturation and precipitation occurs. Material : Fresh urine Tool : Test tube, Pastor's pipette, Math pipette 5 ml : Reagents Acetic acid 6 % Ways of working a) Put clear urine (centrifuge first) into the test tube until it is 2/3 full b) By holding the test tube at the bottom end with test tube clamps, the top layer of urine is heated over a flame until it boils for 30 seconds. c) Pay attention to whether or not there is turbidity in the top layer. If turbidity occurs, it may be caused by protein, calcium phosphate, calcium carbonate. d) Drop 3-5 drops of 6% acetic acid into the still hot urine. If the turbidity is caused by calcium phosphate then the turbidity will disappear. If the turbidity is caused by calcium carbonate then the turbidity will still disappear but with the formation of gas. If the turbidity persists or becomes cloudier, the protein test is positive.Interpretation of Results: - : no turbidity + : light turbidity (like clouds) without grains (protein content 0.01-0.05%) + +: turbidity is easy to see and visible grains in the turbidity (0.05- 0.2%) + + +: urine is clearly cloudy and the turbidity is in pieces (0.2-0.5%) + + + +: urine is very cloudy and has large pieces or clumps (>0.5%) Condition = urine used for examination must be clear. If it is not clear, then centrifugation must be carried out and the supernatant is used. 3. Esbach proteins Method : According to Esbach Principle : Esbach's reagent forms a precipitate with protein contained in the urine and the precipitate is read on a scale on the Esbach tube expressed in promil. Material : 24 hour urine Tools and Reagents Tool : - Albuminometer tube: Reagents : Esbach Reagentm(As. Picrat 10 gr, As. Citrate 20 gr, add distilled water 1000 ml) Ways of working 1. Prepare a clean and dry albuminometer tube and pay attention to whether the scale numbers are clear or not. 2. Take enough urine then add vinegar until the pH is - 6 then filter it. (check with pH paper) 3. Fill the albuminometer tube with urine up to the U mark and reagent to the R mark, then shake until homogeneous 4. Cover with a tube lid or cork and place in a closed place and store for 24 hours at room temperature. Calculation : Total protein in 24 hours = vol. 24 hour urine X result (gram/L) = …….. grams/ 24 hours