ESTIMATION OF PROTEINS AND

LACTOSE IN MILK.

376BCH
Nov 2008
 OBJECTIVES

 To estimate the total protein content of milk.

 To estimate the lactose concentration in milk.
 
INTRODUCTION

 major constituents of milk are lactose, fats and proteins

 The proteins include casein (a phosphoprotein and 80%
of total protein), lactoalbumin and a little lactoglobulin
 There are also important amounts
 Calcium
 phosphorus
 vitamin A , B2 B1, C , D3 and or iron
INTRODUCTION CONT..,
 The white colour of milk is due to emulsified
liquid and to the calcium salt of casein.
 The fat content of human milk is similar to that
of cow’s milk but there is more lactose and less
protein, calcium and phosphorus
INTRODUCTION CONT..,

 Colostrum is more yellowish and contains about twice as

much protein including immunoglobulins

 The protein content falls with time reaching average

levels after about one month. Fat and lactose change
little.
PRINCIPLE ( CASEIN )
 Total protein in milk is estimated as casein by
the Biuret reaction.
 Under strongly alkaline conditions, such
compounds give violet coloured complexes with
cupric salts, which may be
spectrophotometrically estimated.
PRINCIPE ( LACTOSE )
 The diluted milk is added to sodium sulphate-copper
sulphate solution and protein precipitated by the addition of
sodium tungstate solution. The mixture is centrifuged, and
the supernatant liquid is then added to alkaline tartrate.
 Lactose present reduces the copper sulphate to cuprous
oxide.
 Addition of phosphomolybdic acid solution oxidizes the
precipitated cuprous oxide to cupric oxide and is itself
reduced to molybdenum blue.
 The absorbance of the resulting blue colour is taken at 680
nm. From this the concentration of lactose in the diluted
milk may be found if parallel tests using standard lactose
solutions are carried out.