Laboratory Course

Basic Biochemical methods

1. Orientation - Units in biochemistry, calibration of volumetric glassware, introduction to biochemical instrumentation. Preparation of standard solutions. Calibration graphs, methods of plotting data. A typical colorimetric estimation such as Biuret method and proteins,    preparation of buffers. Use of pH meters.
2. Qualitative test for amino acids, carbohydrates and lipids.                            
3. Spectrophotometry -                                         

• Determination of molar extintion coefficients of NAD, NADH, tyrosine, tryptophan, adenine, etc.
• Spectrophotometric estimation of amino acids by ninhydrin method.
• Spectrophotometric estimation of proteins by biuret method.                                                            

        4. Chromatography-                                         

• Separation and identification of sugar by ascending and descending paper chromatography.
• Separation and identification of amino acids by ascending and descending chromatography.
• Separation of lipids, purines, pyrimidines and their quantitation by TLC.
• Quantitative separation of amino acids, nucleosides using Dowex 1 and Dowex 50 resins by ion exchange chromatography. (Practice exercise)
• Extraction and column chromatography of plant pigments( Practice exercise)

        5.    Electrophoresis -                                        

•  Separation of amino acid, proteins by experiments involving electrophoretic techniques.    
• Separation of blue dextran and cobalt chloride by experiments involving electrophoretic techniques.

Clinical Biochemistry     

        6.    Determination of hemoglobin content in blood -
        Osmotic fragility, PCV, ESR, differential counts.
         7.    Determination of blood-glucose -

a.    Hagedorn-Jenesen methods
b.    Nelson-Somogyi method and
c.    glucose oxidase method.                      

                 
      8.    Determination of SGOT, SGPT, bilirubin and calcium in serum/milk powder.
      9.    Determination of urinary creatine and creatinine.
     10.    Estimation of Vitamin 'C' in citrus fruits (titrimetric or colorimetric methods).

Study of Minerals
     11. Determination of Na and K by flame photometry.
      12. Determination of Mg in biological samples