Presented

by
Y.Narayudu
Analysis of milk

What is milk ?
Normal mammary gland secretion of female
mammals
It is the first food for the baby mammaline



Freezing point 0
0
C(water) / -0.55
0
C(solids)

RIBOFLAVIN
Ca.caseinate
Carotene&xanthophyl
Composition of milk
Buffalo milk : high fat content (7.44%)
Cow milk : low fat content ( 3.66%)
TYPES OF MILK
o Standardized milk: buffalo milk & skimmed milk
( fat -4.5% & SNF is 8.5%)

o Whole milk: 3.25% milk fat & 8.25% milk solids
(50% of its calories 4m fat)

o Reduced-fat milk (2%): This milk contains 2% milk fat
(35% of its calories)

o Low-fat milk (1%): 23% of its calories from fat


o Skimmed milk/non-fat milk: NMT 0.5% milk fat 5% of its
calories from fat. Skimmed milk has about half the
calories of whole milk.

o Pasteurized : milk kill bacteria(not spores)
o Pasteurized milk will keep fresh for 2-3 days in a fridge

o Unpasteurized - raw or untreated milk
o It is recommended that babies, young children, the
elderly, pregnant women and anyone with an impaired
immune system should avoid drinking unpasteurized
milk.

63
0
C for 30min
72
0
C For 15sec


o Long - life - milk pasteurized & homogenized and then
kept at a high temp for destroy bacteria.
odd burnt caramel flavour
stored for 1 week


o UHT( ultra-heat treatment) milk heated at high temp
(132
0
C / 270
0
F)
Stored for upto 3 months


o Dried Milk: in powdered form.

o Evaporated : homogenized milk with
considerably reduced water content

o Condensed milk :simply evaporated milk to
which sugar has been added to thicken and
sweeten it. It is mainly used for making
desserts and sweets.

146 calories

49% Fat

30% carbhydrates

21% protein


Cup of milk
IS IT MILK IS CONTAMINATED.?
Contamination of milk
THE NATIONAL NEWS(11-1-2012)& BBC:
68% of Indian milk contaminated
diluted with water
sweeteners
Fat se volume
non-edible solids
glucose and
skimmed milk powder




o Addition of water not only reduces the nutritional
value of milk but contaminated water may also
pose health risks


o The presence of detergent "indicates lack of
hygiene and sanitation in the milk handling





Method of analysis of milk

Preparation of sample

o Warm the sample to 37

0
40
0
C by transferring it to the
beaker & keep it in a water bath maintained at 40
0
- 45
0
C

o Stir slowly for proper homogenisation. Mix sample
thoroughly by pouring back into the bottle, mixing to
dislodge any residual fat sticking to the sides and pour it
back in the beaker.

o Allow the sample to come to room temperature
(26
0
- 28
0
C) and withdraw immediately for analysis.
SPECIFICGRAVITY

o LACTOMETER

o 1.025-1.035 (25
0
C-35
0
C)

o After 12hr of milking rise
0.0013
DETERMINATION OF P
H
o P
H
Meter

o Calibrate the P
H
Meter

o Avg . P
H
6.6

o Due to lactic acid


Determination of total solid:
o take a weight of crucible.

o weigh 5 g of milk in a crucible

o put a crucible in a water bath until dryness.

o after complete dryness put the crucible in an oven, and
weigh after cooling.

o determination the percent of total solid.

%Of total solid = (wt of crucible +sample) after drying wt
of crucible/ wt of sample * 100
RICHMONDS Formula For Total Solids :




For cow milk 0.66
Where,
D = Density
F = % Fat
T=0.25D+1.22F+0.72
DETERMINATION OF CHLORIDE CONTENT
10 ml milk + 40 ml water

add 10 drops of pot.chromate

Titrate with 0.1 N AgN0
3





1 ml of o.1 N AgN0
3
equivalent to 3.55mg of Cl
-
Brick red ppt
INDICATIVE OF DISEASED STATE OF ANIMAL
TITRABLE ACIDITY

10 ml milk + 1 ml phnolpthalein indicator



Titrate with 0.1N NaOH
1 ml 0.1 N NaOH 0.009 g of lactic acid
Determination of Fat in Milk
Gerber Method:
Principle:
o milk +H
2
S0
4
+ iso-amyl
alcohol
o permitting dissol
n
of the
protein and release of fat.
o The tubes are centrifuged
and the fat rising into the
calibrated part of the tube
is measured as percentage
of the fat content of the
milk sample
Procedure:

o 10 ml of H
2
S0
4
into a butyrometer tube & Mix the milk sample

o Add 1 ml of Amyl alcohol,close with a lock stopper

o shake until homogeneous sol
n
.Keep in a water
bath for 5 min at 65
o
C

o centrifuge for 4 min. at 1100 rpm.

o Remove the butyrometer tubes and place in water bath for 5
min.at 65
0
C.

o Read the percentage of fat
Werner Schmidt Method(by Acid Digestion Method):

PRINCIPLE:

o Milk proteins are digested with conc. HCl

o Liberated fat is extracted with alcohol, ethyl ether &
petroleum ether

o Ethers are evaporated

o Residue left behind is weighed to calculate the fat
content.
10 g milk+10 ml conc.HCl

stir with a glass rod until the contents turn dark brown

Mojonnier fat extraction flask

10 ml of C
2
H
5
0H+ 25 ml of ethyl ether

25 ml of petroleum ether
Shake vigorously for 1 min
Centrifuge Mojonnier flask at about 600 rpm
heat on a Bunsen burner
cool to room temp
Shake vigorously for 1 min

Decant the ether sol
n


Repeat extraction

Evaporate the solvent


Dry the fat in oven

Weigh

Calculation:
100 (W1 - W2)
Fat, percent w/w ----------------------
W3
Where
W1 = Wt in g of contents in the flask before removal of fat.

W2 = Wt in g of contents in the flask after removal of fat

W3 = Wt in g of material taken for the test.
Detection of Adulterants in Milk
Detection of Cane Sugar in Milk
Modified Seliwanoff Method:
PRINCIPLE:
Fructose + resorcinol in HCl red colour
procedure
Procedure:
milk + conc. HCl filter

1 ml filtered milk serum &
5 ml modified resorcinol - HCl reagent
Withdraw the tube &observe the colour

red colour

std for 10 min
water bath for exactly 1 min
Test for QAC (Detergents)
o To a centrifuge tube add 1 ml milk, 5 ml water, 1 ml EOSIN
sol
n
& 0.2 ml buffer and shake hard for 10 sec.

o Centrifuge for 5 min at 3200 rpm.

o If QAC is present the bottom layer assumes a red or pink
colour.

o Samples containing 1 mg / kg of QAC show a faint pink

o If the colour is deep pink or red, the amt of QAC can be
approx. determined by titration with a std anionic
detergent sol
n
Detection of added Urea in Milk
o 5 ml of milk is mixed with 5 ml of 1.6 % of
p Dimethyl amino benzaldehyde (DMAB) is
added observed in milk
containing added urea.

o The control (normal milk) shows a slight yellow
colour due to presence of natural urea.
o :

Distinct yellow colour
Estimation of Urea in Milk
Prep
n
of standard Curve:
o Pipette 5 ml std sol
n
s into 25 ml T.T Add 5 ml
DMAB sol
n
to each.

o Prepare reagent blank of 5 ml buffer and 5 ml
DMAB sol
n
. Shake tubes thoroughly and let
stand for 10min.

o Read a@420 nm
Preparation of sample:
o 10 ml of milk sample add 10 ml of Trichloro
acetic acid (TCA) to ppt the proteins and
filtered.
o 5 ml of filtrate + 5 ml of DMAB
o The optical density of the yellow colour is
measured @ 420 nm.
o From standard curve the amount of urea in
milk is calculated.
70 mg per 100 ml (700 ppm)
Detection of preservatives
Hehners Test (HCHO):
2 ml milk in T.T

2 ml of 90 % H
2
SO
4
traces of FeCl
3




Formaldehyde
purple color ring at the junction











Test for presence of Salicylic acid:


50ml milk + 5 ml of dil.HCl + 50 ml ether


Wash ether layer with water


evaporate ether


1 drop of 0.5 % (v/v) FeCl
3






Violet colour
Test for presence of H
2
O
2
Milk + conc.HCl

Mix well

drop of HCHO sol
n

60
0
C

place starch-Iodine paper into sol
n



oxidesation of iodine





BLUE COLOR
Normal flora of milk:
Bacteriological Examination of Milk
oEnterococcus faecalis
oStreptoccus lactus
oLactobacillus sp.
o Candida albicans (yogurt)
Determination of viable bacterial count :


The pour plate method:

After preparation of 10 fold serial dilution from
the milk sample with ringer solution

Using 10 fold serial dilution method
Viable Bacterial Count
9 ml Saline
1 2 3
Milk sample
1 ml milk 1 ml
1 ml
1/10
1/10 x 1/10
1/100
1/100 x 1/10
1/1000
1 ml 1 ml 1 ml Melted NA
1 2 3
Results:
Dilution
factor 1 2

3

X

X . y
10 x
1
X
1
.y
1
10
2
x
2
X
2
.y
2

10
3


x
3
X
3
.y
3
Viable Bacterial Count
No. of colonies per plate
Y
No. of cells per 1 ml =
X
1
.y
1
+ X
2
.y
2
+ X
3
.y
3
3
Results:

o Permissible number of bacterial flora in
pasteurized milk is 5 x 10
4
cfu/ml


o Permissible number of bacterial flora in long
life milk is 10 cfu/ml
Methylene Blue Reduction Test:

o Increasing the number of bacterial flora
will reduce the colour of methylene blue
more rapidly due to increasing consumption
of oxygen.
o i.e: The speed of reduction of methylene
blue colour is directly proportional to the
number of bacteria present in milk sample.
determine quality of the milk
Results:
The shorter the decolorization time, the higher
the number of bacterial flora present in milk, and
the poor quality of milk
Decolorization time Result
30 min 2 hrs Poor quality
2 6 hrs fair quality
6 8 hrs good quality
Over 8 hrs excellent quality
Test for coliforms:


o Done by inoculation of MacConkeys broth with
0.1 ml of milk sample.

o Examine for the production of acid detected by
changing the color of the medium from purple to
yellow.