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Presented By : Mohammad Mursalin
Principle
 When formaldehyde added to a milk sample, Amino
group of amino acid of milk protein binds with
formaldehyde and leaving the carboxyl group free.
 This can set free H+ ions which can be estimated
by titration with standard alkali.
Formaldehyde Amino Acid + Formaldehyde
 Reagents and material required
 Burette
 Pipette
 Beaker
 Formalin (40%) neutral, obtaining by adding a small amount of NaOH, using 2 drops of
phenolphthalein as an indicator.
 Potassium oxalate solution (neutralized to phenolphthalein)
 NaOH solution 0.1M
 Phenolphthalein indicator (0.5% in ethyl alcohol)
 Procedure
 10 ml milk sample pipette out in a dry conical flask.
 Add 2 drops of phenolphthalein
 Add 0.4 ml potassium oxalate (it binds calcium ions) to
encounter the disturbing effects of calcium of milk.
 Set aside for 2 minutes
 Titrated with 0.1M NaOH till end point achieved, note down
burette reading (V1).
 2 ml neutral formalin added, mixed well, diluted with 10
ml CO2 free water.
 1 ml phenolphthalein added.
 Titrated with 0.1M NaOH till end point achieved, note down
burette reading (V2).
 Observation
 % True Protein = V1 x 1.7
 % Casein = V2 x 1.38
THANK YOU

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Pynes method for protein determination in milk sample

  • 1. Presented By : Mohammad Mursalin
  • 2. Principle  When formaldehyde added to a milk sample, Amino group of amino acid of milk protein binds with formaldehyde and leaving the carboxyl group free.  This can set free H+ ions which can be estimated by titration with standard alkali.
  • 3. Formaldehyde Amino Acid + Formaldehyde
  • 4.  Reagents and material required  Burette  Pipette  Beaker  Formalin (40%) neutral, obtaining by adding a small amount of NaOH, using 2 drops of phenolphthalein as an indicator.  Potassium oxalate solution (neutralized to phenolphthalein)  NaOH solution 0.1M  Phenolphthalein indicator (0.5% in ethyl alcohol)
  • 5.  Procedure  10 ml milk sample pipette out in a dry conical flask.  Add 2 drops of phenolphthalein  Add 0.4 ml potassium oxalate (it binds calcium ions) to encounter the disturbing effects of calcium of milk.  Set aside for 2 minutes  Titrated with 0.1M NaOH till end point achieved, note down burette reading (V1).  2 ml neutral formalin added, mixed well, diluted with 10 ml CO2 free water.  1 ml phenolphthalein added.  Titrated with 0.1M NaOH till end point achieved, note down burette reading (V2).
  • 6.  Observation  % True Protein = V1 x 1.7  % Casein = V2 x 1.38