![Page 1: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/1.jpg)
Serum biochemical parametersTOTAL Protein assay
Biochemistry Clinical practiceCLS 432
Dr. Samah KotbLecturer of Biochemistry
2015
![Page 2: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/2.jpg)
What do you know about liver function
![Page 3: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/3.jpg)
• 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
![Page 4: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/4.jpg)
• Total protein was estimated according to the method of (Burtis A and Koller A ., 1999) , Colorimetric method.
Principle
![Page 5: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/5.jpg)
• Proteins give an intensive violet –blue complex with copper salts in an alkaline medium. Iodide is included as an antioxidant.
Principle
![Page 6: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/6.jpg)
• The intensity of the color formed is proportional to the total protein concentration in the sample.
![Page 7: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/7.jpg)
• 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
![Page 8: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/8.jpg)
• 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
![Page 9: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/9.jpg)
• 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.
![Page 10: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/10.jpg)
• 3- The contents were mixed well, incubated at 37°C for 5min, standard and blank tubes.
![Page 11: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/11.jpg)
• 4- The absorbance read at 540 nm.
• 5- Blank was used for zero adjustment of standard, and the test.
![Page 12: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/12.jpg)
• A sample • Concentration of protein ( g/dL) = x 7.0 • Where: A standard • A: absorbance of recorded sample• 7.0: Standard concentration
Calculation:
![Page 13: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/13.jpg)
• 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
![Page 14: Serum biochemical parameters TOTAL Protein assay Biochemistry Clinical practice CLS 432 Dr. Samah Kotb Lecturer of Biochemistry 2015](https://reader035.vdocument.in/reader035/viewer/2022062301/5697bfa01a28abf838c9527a/html5/thumbnails/14.jpg)