1. Technique in Biochemistry 1 Techniqe in Biochemistry

2. Technique in Biochemistry is common analytical technique used in clinical. The two major technique in biochemistry : 1. Qualitative technique Structure of biomolecule To detect the pathological conditions 2. Quantitative technique Concentration or level of biomolecule Result of the technique can use for : 1. Diagnostic 2. Prognostic22 3. Therapy Techniqe in Biochemistry 2 Technique in Biochemistry

3. Biomolecules: Proteins Carbohydrates Lipids Nucleic acids General Techniques: Separation and purification of biomolecules Characterizations of biomolecules Techniqe in Biochemistry 3 Biomolecules & General Techniques

4. Techniqe in Biochemistry 4 Quantitative analysis Carbohydrates Glucose Glucose blood level  diabetes mellitus Technique analysis : enzymatic Reaction : glucose + O2 + H2O gluconic acid + H2O2 H2O2 + Phenol + 4-aminophenazone H2O + Quinone dye (red color) Glucoselevel is measured by spectrophotometer Absorbance at 505 nm due to the formation of the BCG-Albumin Glucose oxydase Peroxydase

5. From serum or plasma Technique analysis : enzymatic Albumin + BCG BCG-Albumin complex BCG is sodium salt (Bromocresol green) Complex BCG-Albumin complex is directly proportional to the concentration of albumin in the sample. Albumin level is measured by spectrophotometer Absorbance at 630 nm due to the formation of the BCG-Albumin Techniqe in Biochemistry 5 Protein Albumin

6. Techniqe in Biochemistry 6 Lipids Cholesterol Blood cholesterol Technique analysis : enzymatic Reaction : Cholesterol ester cholesterol + fatty acid Cholesterol + O2 4 cholesterol + H2O2 H2O2 + 4-aminophenazone + Phenol 4-(p-benzokinon-monoamino)phenazone + H2O cholesterollevel is measured by spectrophotometer Absorbance at 546 nm Cholesterol esterase Cholesterol oksidase peroksidase

7. Techniqe in Biochemistry 7 Lipids Triglyceride Blood triglyceride Technique analysis : enzymatic Reaction : Triglyceride glycerol + free fatty acid Glycerol + ATP glycerol + 4 phosphate + ADP H2O2 + 4 aminoantipyrin (4AAP)+ 3,5-dicholoro-2- dihydroxybenzenesulfonic acid (DHBS) Quinoneimine dye Red color + H2O cholesterol level is measured by spectrophotometer Absorbance at 546 nm lipoprotein lipase Glicerol Kinase peroksidase

8. Techniqe in Biochemistry 8 Quantitative analysis Albumin Plasma protein electrophoresis on cellulose acetate

9. Isolation DNA  lab act week 4 PCR  in research Techniqe in Biochemistry 9 DNA

10. Reference Murray R.K., Harper’s Illustrated Biochemistry ST-reagensia, Enzyme assay methods 10 Techniqe in Biochemistry