VPB 112: General Veterinary Biochemistry (1+1)

PREPARATION OF BUFFER - PROCEDURE

• Preparation of 500 ml of 0.1N Acetic acid
  • The commercially available acetic acid is approximately 17.6 N. Therefore, 2.85 ml of acetic acid is to be taken in a beaker and diluted up to 500 ml with distilled water.
  • The solution is stirred well and this is 500 ml of 0.1 N Acetic acid solution (CH₃COOH).
• Preparation of 500 ml of 0.1N Sodium Acetate solution
  • Molecular weight of CH₃COONa.5H₂O = 136 g.
  • Therefore, weigh 6.8 g of Sodium acetate in a watch glass. This is dissolved in distilled water and the volume made up o 500 ml. This solution gives 500 ml of 0.1 N CH₃COONa.
• Preparation of different concentrations of Acetate buffer solution
  • Various concentrations of acetate buffers (Total Volume = 90 ml each) to be prepared are mentioned in the protocol given below, wherein different volumes of 0.1N acetic acid and 0.1 N sodium acetate prepared above are to be mixed and stirred well.

Test Tube No. Reagents	I (ml)	II (ml)	III(ml)	IV (ml)	V(ml)	VI (Control) (ml)
1. 0.1N CH3COONa	10	30	50	70	90	Only D.W 90
2. 0.1 N CH3COOH	80	60	40	20	10
Total Volume of Acetate Buffer prepared	90	90	90	90	90	90