Understanding Nutrition

Al-Quds University Body Fluid Lab. Report Chemical Examination of Urine Prepared By Lucia Principles Benedicts Test for Reducing Sugars Urinary sugars when boiled in Benedicts reagent reduce coppersulphateto a ruddy cuprous oxide precipitate in hot alcalescent medium, the intensity of which is proportional to the amount of sugar pledge in the weewee. The results are reported as 1+,2+, etc. depending upon thecolourand intensity of the cuprous oxide precipitate. turbidimetric mode protein in piddle Quantitative Tests for 24-Hour Specimens. Trichloroacetic pane (TCA) prove.The addition of TCA to a urine specimen precipitates the protein in a fine suspension that is quantified spectrophotometrically at 420 nm (nanometers) by comparison with a similarly treated standard. passion and acetic acerbic method Heat test Based on the principle of heat coagulation and precipitation of proteins. If any turbidity appears, add 2 drops of 33% acetic acid. (Acidification is necessary be cause in alkaline medium heating may precipitate phosphates). If the precipitate is payable to proteins, it entrust increase on acidification and if it is due to phosphates, it will dissolve again.Sulphosalicylicacid method Urine Protein Sulfosalicylic Acid Precipitation Test (SSA) Principle three percent (3%) Sulfosalicylic Acid (SSA reagent) is added to a small and equal volume of clear urine. The acidification causes precipitation of protein in the adjudicate (seen as increasing turbidity), which is subjectively graded as trace,1+, 2+, 3+ or 4+. Bence J angiotensin converting enzymes protein Bence Jones chemical reaction involves heating urine to 140F (60C). At this temperature, the Bence Jones proteins will clump. The clumping disappears if the urine is further heated to boil and reappears when the urine is cooled.Other clumping procedures using salts, acids, and other chemicals are also utilize to detect these proteins. These types of test will go against whether or not Bence Jones proteins are present, nevertheless not how much is present. haemosiderin Hemosiderin stain is use to indicate the front end of iron storage granules called hemosiderinby microscopic examination of urine sediment. Granules of hemosiderin stain blue when potassium ferrocyanide is added to the sample. The Prussian blue stain may also be used to identify siderocytes (iron-containing red blood cells RBCs) in peripheral blood.The presence of siderocytes in circulating RBCs is abnormal. urobilinogen This test is based on a modified Ehrlich reaction in which p-diethylaminobenzaldehyde reacts with urobilinogen in a powerfully acid medium. Colors range from light pink to bright magenta. Results * Benedicts Test result for cup G3 4+ Brown color appear. * Turbidimetric method result for cup of 24-hrs urine Tube Absorbance Test+test-blank 0. 058 Standard+ irrigate blank 0. 010 Calculation Total protein (mg/dl)=At/Ast ? conc. St 0. 058/0. 010 ? 100=580 mg/dl Total protien(mg/24 hrs) =urine protein (mg/dl)? urine volume(ml)/100 = 580? 2000/100)=11600 mg /dl customary values 0-150 mg/24 hrs * Heat and acetic acid method and Sulphosalicylicacid method for cup P4 4+ precipitation appeared. * Bence Jones protein for cup P4 Clear by and by 15 min of boiling so negative for Bence Jones protein. * Hemosiderin results Few Hemosiderin granules was seen under Microscope * Urobilinogen result for cup G3Negative result (no appearance of red color). Interpretation Benedicts Test for Reducing Sugars Normal urine does not contain any reducing sugar. If protein is present in large amounts, it may intercede with the precipitation of the cuprous oxide.To overcome this problem, precipitate the proteins using 3% SSA fall into place using aWhatmanfilter paper and use the filtrate to test the amount of sugar present. As a lineament look into measure, standards containing known amounts of glucose are prepared in saturated benzoic acid and one of the standards is used all day to check the reliability of the patients results. The standard results may be transformed in the following semi-quantitative way. turbidimetric method protein in urine For turbidimetric methods, there were no apparent problems of comparative bias betwixt human albumin and serum-based materials and urines used in this study.Perhaps this was because all materials were diluted in 9 g/L saline turbidimetric methods generally suffer from failure of standards and samples to form precipitates identically,and precipitation may not occur at low protein concentrations in urines of high ionic strength. Heat and acetic acid method This test is raw enough to detect protein down to a concentration of 2-3 mg%. Ifan alkalineurine is boiled, the protein may be converted into the so- called alkalinemetaprotein, which is not coagulated by heat. Therefore it is always better to acidify the urine before doing this test.If too much acetic acid is added, the protein may be converted to the so-ca lled acidmetaprotein, which is also not coagulated by heat. Therefore the urine should be only mildly acidic. Sulphosalicylicacid method Thesulphosalicylicacid method will not detect protein ina normalurine, but will be sensitive enough to detect protein present down to 20mg%. As a quality control measure, a 22g/dl albumin solution can be diluted appropriately with 0. 9 g/dl sodium chloride to get standards containing 20, 50, 200, 500 and 2500 mg/dl proteins.These standards are stable for one month when stored at 2-80C. Bence Jones protein Monoclonal light chain proteinuria (Bence Jones proteinuria) is seen in patients with light chain myeloma, in approximately 50% of those with IgG and IgA myeloma, and in some patients with other lymphoproliferative disorders (eg, macroglobulinaemia) and plasma cell dyscrasias (amyloid). good paraprotein molecules may also be detected in serum. Urine protein dipsticks do not detect Bence-Jones protein. Hemosiderin Hemosiderin is present in disease s involving a true siderosis of kidney parenchyma (hemochromatosis).It is also present 2-3 days after an swell hemolytic episode that produces hemoglobinemia and hemoglobinuria. Hemosiderin granules are found in intact renal tubelike epithelial cells or occasionally in casts and may also be seen extracellularly. Urobilinogen Interpretation of results will depend upon several factors the variability of color perception the presence or absence of inhibitory factors the presence or absence of inhibitory factors typically found in urine, the specific gravity or the pH and the lighting conditions under which the product is used.