Upload
eugene-peregrine-hicks
View
226
Download
2
Tags:
Embed Size (px)
Citation preview
Serum biochemical parametersTOTAL Protein assay
Biochemistry Clinical practiceCLS 432
Dr. Samah KotbLecturer of Biochemistry
2015
What do you know about liver function
• 1. Total and conjugated bilirubin • 2. ALT• 3. AST• 4. Total protein• 5. GGT• 6. ALP • 7. 5 -nucleotidase′• 8. Albumin• 9. prothrombin time
Liver Function tests
• Total protein was estimated according to the method of (Burtis A and Koller A ., 1999) , Colorimetric method.
Principle
• Proteins give an intensive violet –blue complex with copper salts in an alkaline medium. Iodide is included as an antioxidant.
Principle
• The intensity of the color formed is proportional to the total protein concentration in the sample.
• Total protein Standard : Bovine serum albumin standard (7g/dL)(70 g/L).
• Biuret reagent: R1
• Sodium Potassium tartrate (21 mmol/L)
• Sodium hydroxide (0.75 mol/L)
• Potassium iodide (6 mmol/L)
• Cupric Sulfate (6 mmol/L)
Reagents
• 1- 1 ml of Biuret reagent was added into both unknown test and control tubes, then 0.02 ml of serum were added to unknown.
Procedure
• 2- 0.02 ml of Bovine albumin primary standard and 1 ml of Biuret reagent were added into standard tube, 1 ml of Biuret reagent was added into blank tube.
• 3- The contents were mixed well, incubated at 37°C for 5min, standard and blank tubes.
• 4- The absorbance read at 540 nm.
• 5- Blank was used for zero adjustment of standard, and the test.
• A sample • Concentration of protein ( g/dL) = x 7.0 • Where: A standard • A: absorbance of recorded sample• 7.0: Standard concentration
Calculation:
• Adults: 6.6-8.7 g/dL (66-87g/L).• Prematures: 3.6-6.0 g/dL (36-60 g/L).• Newborns : 5.3-8.9 g/dL (53-89 g/L)• Pregnancy: Concentration lowers from 69 to
61 g/L
Reference values