Milk composition

DR SHAMAS MURTAZA
 Definition of Milk
2

 Milk may be defined as the whole, fresh, clean, lacteal

secretion obtained by the complete milking of one or more
healthy milch animals, excluding that obtained within 15
days before or 5 days after calving or such periods as may
be necessary to render the milk practically colostrums-
free, and containing the minimum prescribed percentage
of milk fat and milk solid not fat.
 MILK
Total Solids
Water

Fat (Lipids) SNF

Emulsion form (50-
100 nm dia.)

Lactose Mineral Other

Nitrogen
(solution form matter constituents
True fat ( 98% TGs + Associated Substance
MG+ DG+ FFA Substance .01 -1 nm)
PO4, citrates ,
Chlorides of Na, K,
Ca, Mg + traces of
Phospholipids Vitamins Fe, Cu, I etc.
(Lecithin, Cephalin, Cholesterol Carotene
(A,D,E,K)
Sphyngomylin)
•Pigments
•Dissolved Gases
•Vit. C &
Protein B Complex.
Non protein (Suspension, 1-100nm dia.)
•Enzymes etc

Caseins 3
(α,β,γ,κ) β Lactglobulin α- Lactalbumin Proteose Peptones
 Milk Composition
4

 The principal constituents of milk are water, fat,

proteins, lactose (milk sugar) and minerals (salts).
 Milk also contains trace amounts of other substances
such as pigments, enzymes, vitamins, phospholipids
(substances with fatlike properties), and gases.
❑The residue left when water and gases are removed is
called the dry matter (DM) or total solids content of the
milk
Composition of Milk from Various Animals
5
6
 Milk Fat
7

 The milk fat exists as small globules or droplets dispersed

in the milk serum.
 Their diameters range from 0.1-20µm (1µm = 0.001 mm)
 The average size is 3-4 µm and there are some 15 billion
globules per ml.
 The emulsion is stabilized by a very thin membrane only
5-10 nm thick (1 nm=10-9m) which surrounds the
globules and has complicated composition.
Composition of Milk Fat
8
 Conti…
9

Milk fat consists of

 Triglycerides (Dominating components)
 Di and Monoglycerides
 Fatty acids,
 Sterols,
 Carotenoids (giving the yellow color of the fat)
 Vitamins (A, D, E and K)
 Conti…
10
The fat globule membrane consists of
 Phospholipids
 Lipoproteins
 Cerebrosides
 Proteins
 Nucleic acids
 Enzymes
 Trace elements (metals) and bound water.
➢ Fat globules have the lightest density at 15.5°C 0.93
g/cm3, they tend to rise to the surface when milk is left to
stand in a vessel.
 Conti…
11
❑The rate of rise follows Stokes’ Law, but the small size of
the fat globules makes creaming a slow process.
❑Cream separation can be accelerated by aggregation of fat
globules under the influence of a protein called
agglutinin.
 These aggregates rise much faster than individual fat
globules.
 The aggregates are easily broken up by heating or
mechanical treatment.
❑ Agglutinin is denatured at time-temperature combinations
such as 65°C/10 min or 75°C/2 min and the possibility of
aggregation disappears.
Principal Fatty Acid in Milk
12
 Fatty Acid in Milk
13

 Four most abundant fatty acids in milk are

1. Myristic,
2. Palmitic
3. Stearic and
4. Oleic acids
 This variation of fatty acids affects the hardness of the fat.
 Fat with a high content of high-melting fatty acids, such as
palmitic acid will be hard.
 Fat with a high content of low-melting oleic acid makes
soft butter.
 Iodine value
14

 The iodine value states the percentage of iodine that the fat
can bind.
 Iodine is taken up by the double bonds of the unsaturated
fatty acids.
 Since oleic acid is by far the most abundant of the
unsaturated fatty acids, which are liquid at room
temperature the iodine value is largely a measure of the
oleic-acid content and thereby of the softness of the fat.
 The iodine value of butterfat normally varies between 24
and 46.
 Refractive index
15

 The amount of different fatty acids in fat also affects the

way it refracts light.
 This is a quick method of assessing the hardness of the
fat.
 The refractive index normally varies between 40 and 46.
 Cow milk proteins
Concentration g/kg %
Total proteins 33.0 100.0
Total caseins 26.0 79.5
as1-casein 10.0 30.6
as2-casein 2.6 8.0
b-casein 9.3 28.4
-casein 3.3 10.1

Whey proteins 6.3 19.3

a-lactalbumin 1.2 3.7
b-lactoglobulin 3.2 9.8
Bovine Serum Albumin 0.4 1.2
Immunoglobulins 0.7 2.1
Other (proteoses peptones)
- 0.8 2.4
Proteins of fat globule membrane
16
0.4 1.2
 COMPOSITION OF CASEIN MICELLES
⚫ 93 % caseins : 4 phosphoproteins
⚫ as1-CN : 36 %
⚫ as2-CN : 10 %
⚫ b-CN : 34 %
⚫ -CN : 12 %
⚫ 7 % : colloidal mineral complex containing phosphate,
calcium, magnesium and citrate
In milk the whey proteins are in colloidal solution
and the casein in colloidal suspension.

17
 Amino acids
18

1. Negatively charged in alkaline solutions

2. Neutral at equal + and – charges
3. Positively charged in acid solutions
 If the side chain is polar, the water-attracting properties of the
basic and acid groups, in addition to the polar side chain, will
normally dominate and the whole amino acid will attract water
and dissolve in water. Hydrophilic.
 A long hydrocarbon chain repels water and makes the amino
acid less soluble or compatible with water. Hydrophobic
 If there are certain radicals such as hydroxyl (–OH) or amino
groups (–NH2) in the hydrocarbon chain, its hydrophobic
properties will be modified towards more hydrophilic.
 Conti…
19

 Hydroxyl groups in the chains of some amino acids in

casein may be esterified with phosphoric acid.
 Such groups enable casein to bind calcium ions or
colloidal calcium hydroxyphosphate, forming strong
bridges between or within the molecules.
 The side chains of some amino acids in milk proteins
carry an electric charge which is determined by the pH of
the milk.
 When the pH of milk is changed by addition of an acid or
a base, the charge distribution of the proteins is also
changed.
20
21
22
 Casein
23

 Casein is a group name for the dominant class of proteins

in milk.
 The caseins easily form polymers containing several
identical or different types of molecules.
 Abundance of ionisable groups and hydrophobic and
hydrophilic sites the molecular polymers formed.
 The polymers are built up of hundreds and thousands of
individual molecules and form a colloidal solution
These molecular complexes are known as casein micelles.
 Such micelles may be as large as 0.4 microns and can
only be seen under an electron microscope.
 A medium-sized micelle consists of about 400 to 500 sub-
micelles which are bound together.
24
 Subgroups of casein
25
 The three subgroups of casein, αs-casein, κ-casein and β-
casein,
 All heterogeneous and consist of 2-8 genetic variants.
 Genetic variants of a Protein differ from each other only
by a few amino acids.
 The three Sub- Groups have in common the fact that one
of two amino acids containing hydroxy groups are
esterifies to phosphoric acid.
 The phosphoric acid binds calcium and magnesium and
some of the complex salts to form bonds between and
within molecules.
Casein micelles
26
 Conti…
27

 The content of α-, β- and κ-casein is heterogeneously

distributed in the different micelles.
 Calcium salts of αs-casein and β-casein are almost
insoluble in water, while those of κ-casein are readily
soluble.
 Due to the dominating localisation of κ-casein to the
surface of the micelles, the solubility of calcium κ-
caseinate prevails over the insolubility of the other two
caseins in the micelles,
 So whole micelle is soluble as a colloid.
 Conti…
28

 The α and β-caseins are mainly concentrated in the middle

of the sub-micelles, while κ-casein predominates on the
surface.
 The hydrophilic protruding chain of the κ-casein
protrudes from the surface of the sub-micelles forming a
hairy layer (5-10 nm).
 The κ-casein-deficient sub-micelles are mainly located in
the centre of the micelle,
 whereas the κ-casein-rich sub-micelles predominate on
the surface, giving the whole micelle a hairy surface layer.
 The hairy layer of the κ-casein’s protruding chain is
partially responsible for the micelle’s stability through a
major contribution to the negative charge of the micelles.
 Conti…
29

 The calcium phosphate and hydrophobic interactions

between sub-micelles are responsible for the integrity of
the casein micelles.
 Adding an excess of Ca and phosphate results in
aggregation of sub-micelles into larger units.
 The reason for this aggregation is presumably the
deposition of Ca-phosphate in the sub-micelles, which
lowers their electric charge and makes them more
compact.
 Casein curd
30

 If the hairy layer is removed e.g. by acid addition or

rennet – induced hydrolysis, the colloidal stability of the
micelle is destroyed and the micelles coagulate or
precipitate.
 In an intact micelle there is surplus of negative charges,
therefore they repel each other.
 Water molecules held by the hydrophilic sites of k-casein
form an important part of this balance.
 When the hydrophilic sites are removed, water will start
to leave the structure.
 This gives the attracting forces room to act.
 Precipitation by casein
31

 One characteristic property of casein is its ability to

precipitate.
 Due to the complex nature of the casein molecules, and
that of the micelles formed from them, precipitation can be
caused by many different agents.
 Precipitation by acid
32

 The pH will drop if an acid is added to milk or if acid-

producing bacteria a allowed to grow in milk. This will
change the environment of the casein micelles in two
ways.
1. Firstly colloidal calcium hydroxyphosphate, present in
the casein micelle, will dissolve and form ionised
calcium, which will penetrate the micelle structure and
create strong internal calcium bonds.
2. Secondly the pH of the solution will approach the
isoelectric points of the individual casein species.
 Conti…
33

 Both methods of action initiate a change within the

micelles.
 Growth of the micelles through aggregation and ending
with a more or less dense coagulum.
 The isoelectric points of the casein components depend on
the ions of other kinds present in the solution.
 Theoretical values, valid under certain conditions, are pH
5.1-5.3.
 Precipitation by Enzymes
34

 The amino-acid chain forming the κ-casein molecule

consists of 169 amino acids.
 The bond between amino acids 105 (phenylalanin) and
106 (methionin) is easily accessible to many proteolytic
enzymes.
 The soluble amino end contains amino acids 106 to 169,
which are dominated by polar amino acids and the
carbohydrate, which give this sequence hydrophilic
properties.
 35

 This part of the κ-casein molecule is called the

glycomacro-peptide and is released into the whey in
cheese making.
 The remaining part of the κ-casein, consisting of amino
acids 1 to 105, is insoluble and remains in the curd
together with αs- and β-casein.
 This part is called para-κ-casein.
 The formation of the curd is due to
1. Sudden removal of the Hydrophilic Macropeptides
2. Imbalance in intermolecular forces.
 Conti…
36

 Bonds between hydrophobic sites start to develop and are

enforced by calcium bonds which develop as the water
molecules in the micelles start to leave the structure.
 This process is usually referred to as the phase of
coagulation and syneresis.
 The splitting of the 105-106 bond in the κ-casein
molecule is often called the primary phase of the rennet
action.
 While the phase of coagulation and syneresis is referred to
as the secondary phase.
 Conti…
37

 There is also a tertiary phase of rennet action, where the

rennet attacks the casein components in a more general
way. This occurs during cheese ripening.
 The durations of the three phases are determined mainly
by pH and temperature.
 Whey Proteins
38

 Whey protein is the name commonly applied to milk

serum proteins.
 They are not precipitated at their isoelectric points.
 They are, precipitated by polyelectrolytes such as
carboxymethyl cellulose.
 When milk is heated, some of the whey proteins denature
and form complexes with casein, thereby decreasing the
ability of the casein to be attacked by rennet and to bind
calcium.
 α-lactalbumin
39

 Whey proteins in general, and α-lactalbumin in particular,

have very high nutritional values.
 Their amino acid composition is very close to that which
is regarded as a biological optimum.
 α - Lactalbumin contains 123 amino acids and represents
about 25% of the serum proteins in milk.
 It is present in milk from all mammals and plays a
significant part in the synthesis of lactose in the udder.
 The protein has a very compact, globular structure that is
nearly spherical in shape.
 α - Lactalbumin is the most heat stable serum protein in
milk. 50% of the α -Lactalbumin will not be denatured
even after 30 minutes of heating at 77C.
 β-lactoglobulin
40

 β - Lactoglobulin contains 162 amino acids.

 It is the major milk serum protein. It is about 50% of the
serum protein and 8% of the protein in milk.
 There is no β -Lactoglobulin present in human milk.
 β - Lactoglobulin can be irreversibly denatured by heat.
 This stress causes rupture of intramolecular disulfide
bonds and precipitation
 41
 If milk is heated to over 60 °C, denaturation is initiated
where the reactivity of the sulphur-amino acid of β-
lactoglobulin plays a prominent part.
 Sulphur bridges start to form
1. between the β-lactoglobulin molecules,
2. between β-lactoglobulin molecule and a κ-casein
3. between β-lactoglobulin and α-lactalbumin.
 At high temperatures, sulphurous compounds such as
hydrogen sulphide are gradually released.
 These sulphurous compounds are responsible for the
“cooked” flavour of heat treated milk.
 Immunoglobulins and related minor proteins
42

 Immunoglobulins are antibodies synthesized in response

to stimulation by specific antigens.
 They are specifically present in blood.
 Their content in cows’ milk is low, but some of them are
present in higher levels in colostrum and human milk.
 They can also act against “particles” such as bacteria,
viruses and even fat globules, and flocculate them, a
reaction called agglutination.
 Serum Albumin
43

• Comes from blood

• Role in the transport of bile salts, fatty acids

 Membrane Proteins
44

 Membrane proteins are a group of proteins that form a

protective layer around fat globules to stabilize the
emulsion
 Some of the proteins contain lipid residues and are called
lipoproteins.
 Denatured Proteins
45

 As long as proteins exist in an environment with a temperature

and pH within their limits of tolerance, they retain their
biological functions.
 If they are heated to temperatures above a certain maximum
their structure is altered.
 The same thing happens if proteins are exposed to acids or
bases, to radiation or to violent agitation.
 The proteins are denatured and lose their original solubility.
 When proteins are denatured, their biological activity ceases.
 Enzymes, a class of proteins whose function is to catalyse
reactions, lose this ability when denatured.
 The reason is that certain bonds in the molecule are broken,
changing the structure of the protein.
 After a weak denaturation, proteins can sometimes revert to
their original state, with restoration of their biological
functions.
 Milk Enzymes
46

 Enzymes in milk occur in various states:

1 As unassociated forms in solution,
2 Associated or an integral part of membrane fractions,
such as the fat globule membrane.
3 Associated with casein micelles,
4 As part of the microsomal particles.
 The origin of these enzymes in milk is from:
➢ cow’s udder (synthesized enzymes)
➢ Bacterial enzymes (bacterial source).
 Plasmin
47

 This enzyme hydrolyzes proteins.

 Limited proteolysis of B-casein by this enzyme is
responsible for the presence in milk of large polypeptides
derived from this protein, known as the gamma-caseins.
 Activity of this enzyme is also important in cheese
ripening and the stability of casein micelles in various
products such as UHT milk.
 Nearly, 80% of its proteolytic activity is lost when milk is
pasteurized.
 Microbial derived proteases are more heat stable than
native
 Proteases in milk and they tend to survive even UHT
processing.
 Lactoperoxidases
48

 Peroxidase transfers oxygen from hydrogen peroxide

(H2O2) to other readily oxidisable substances.
 This enzyme is inactivated if the milk is heated to 80°C
for a few seconds,
 Catalase
49

 Catalase splits hydrogen peroxide into water and free

oxygen.
 By determining the amount of oxygen that the enzyme
can release in milk, it is possible to estimate the catalase
content of the milk and learn whether or not the milk has
come from an animal with a healthy udder.
 Milk from diseased udders has a high catalase content,
while fresh milk from a healthy udder contains only an
insignificant amount.
 Catalase is destroyed by heating at 75°C for 60 seconds.
 Phosphatase
50

 Phosphatase split certain phosphoric-acid esters into

phosphoric acid and the corresponding alcohols.
 Phosphatase is destroyed by ordinary Pasteurization (72
°C for 15-20 seconds), so the phosphatase test can be used
to determine whether the Pasteurization temperature has
actually been attained.
 The phosphatase test should preferably be performed
immediately after heat treatment.
 Lipase
51

 Lipase splits fat into glycerol and free fatty acids. Excess free
fatty acids in milk and milk products result in a rancid taste.
 The action of this enzyme seems, in most cases, to be very
weak, though the milk from certain cows may show strong
lipase activity.
 The quantity of lipase in milk is believed to increase towards
the end of the lactation cycle.
 Lipase is, to a great extent, inactivated by pasteurization, but
higher temperatures are required for total inactivation.
 Beneficial effects of its activity include the possible aid in
initial digestion and absorption of milk lipids in the intestinal
tract and flavor in certain cheeses made from raw milk.
 sodium and magnesium tend to stimulate the lipase activity,
while calcium and magnesium show an inhibitory effect.
 Mineral in Milk
52

 Mineral salts occur in solution in milk serum or in casein

compounds.
 The most important salts are those of calcium, sodium,
potassium and magnesium.
 They occur as phosphates, chlorides, citrates and
caseinates.
 Potassium and calcium salts are the most abundant in
normal milk.
 The amounts of salts present are not constant.
 Towards the end of lactation, and even more so in the case
of udder disease, the sodium chloride content increases
and gives the milk a salty taste.
 Mineral composition (total and soluble) of
milk
[Total] – [Soluble] = [Micellar] or [Colloidal]

Mineral Concentration (mg/kg)

Total calcium 1250
Soluble calcium 350
Total magnesium 115
Soluble magnesium 70
Total sodium 425
Soluble sodium 400
Total potassium 1600
Soluble potassium 1500
Total chloride 1100
Soluble chloride 1100
Total phosphorus 950
Soluble phosphorus 420
Total inorganic phosphate (in phosphorus) 720
Soluble inorganic phosphate (in phosphorus) 300
Total citrate 1650
Soluble citrate 1500
53
Partition of Major Minerals in Colloidal and
Soluble Phases (% of Total Minerals)
54
 Carbohydrates in Milk : Two types
55

 Free
• Lactose
• Glucose
• Galactose
• N-acetylated glucose
• N-acetylated galactosamine
 Linked to proteins
• glycoproteins
• -casein
• Lactoferrin
• Lactoperoxidase
 Conti…
56

 Disaccharide composed of one glucose unit and one

galactose unit
 Major sugar of the most milks from different mammals
 Lactose is one of the least soluble of the common sugars,
having solubility in water of only 17.8% at 25◦C
 Cow and human milks contain about 4.8 % and 7 % of
lactose respectively.
 When lactose undergoes dehydration to form lactulose. It
stimulates the growth of Bifidobacterium bifidum and is
thus beneficial in establishing healthy commensal
microbiota in the gut.
 Lactose is a good source of energy and may promote
calcium absorption.
 Conti…
57

 Digestion of lactose presents a problem in some people as

they lack beta-d-galactosidase enzyme in their GIT.
 Consequently, dietary lactose is not hydrolyzed and
reaches colon where it is metabolized by colonic bacteria
forming gases (methionine and hydrogen).
 Accumulation of gas leads to discomfort caused by
bloating and diarrhea.
 Such lactose malabsorption is aggravated by yogurt
containing live cultures,
 the culture furnishes the lactose-hydrolyzing enzyme
beta-d-galactosidase and normal digestion pattern is
restored.
 Role of lactose in milk and dairy products
• Contribute to the nutritive value of milk and dairy
products
• Essential component for the fermentation of some dairy
products (yoghurts, cheeses, …)
• Affects the texture of some concentrated and frozen
products.
•During rapid drying amorphous lactose is formed. This
form of lactose is very hygroscopic and causes caking in
dried products containing moisture levels of 8% or more.
•Participate to the color and flavor changes of dairy
products during heat treatments and storage (Maillard
reaction)
58
 Food Applications
59

 Infantile food
 Glazing, prevent crystallisation of other sugars in
mixture, ... (confectionery - bakery)
 Appearance and taste of bakery products, fried foods...
(Maillard)
 Exhauster of taste (sauces, french dressing,...)
 Stabiliser of aroma
 Agent of encapsulation (confectionery)
 Vitamins in Milk
60

 Vitamins are organic substances that occur in very small

concentrations in both plants and animals.
 They are essential to normal life processes, but
cannot be synthesized by the body.
 Among the best known vitamin in milk are A, the vitamin
B group, vitamin C and D.
 Physical Properties of Milk
61

Appearance
 The opacity of milk is due to its content of suspended
particles of fat, proteins and certain minerals.
Osmotic Pressure
 Osmolality is a measure of the total number of dissolved
particles in a given volume of solution given in osmol/KJ.
 Osmotic pressure is controlled by the number of
molecules or particles, not the weight of solute; thus 100
molecules of size 10 will have 10 times the osmotic
pressure of 10 molecules of size 100
 Conti…
62

Freezing Point
 The freezing point of milk is lower than that of pure water due
to the dissolved components such as lactose and soluble salts.
 The freezing point of milk is the only reliable parameter to
check for adulteration with water.
 The freezing point of milk from individual cows has been
found to vary from –0.54 to –0.59°C.
 Adulterated milk will show increased freezing point due to
lower molal concentration of lactose and salts.
Boiling Point
 The boiling point of milk is higher than that of pure water due
to dissolved components.
 The boiling point of milk is 100.17◦C.
 Conti…
63

Density
 Milk density at 20◦C ranges from 1.027 to 1.033 with an
average of 1.030 g/cm3
 Temperature affects density because milk expands when
heated and so it becomes less dense as temperature rises.
Specific Gravity
 It is the ratio of the mass of a solution or a substance to
the mass of a similar volume of water.
 Fresh whole milk has specific gravity in range of 1.030-
1.035, with an average of 1.032.
 Other Constituents of Milk
64

 Milk always contains somatic cells (white blood

corpuscles or leucocytes).
 The somatic cell content of milk from healthy animals is
as a rule lower than 200 000 cells/ml, but counts of up to
400 000 cells/ml can be accepted.
 Milk also contains gases, some 5-6 % by volume in milk
fresh from the udder, but on arrival at the dairy, the gas
content may be as high as 10 % by volume.
 The gases consist mostly of carbon dioxide, nitrogen and
oxygen.
 Dissolved in the milk
 Bound and non-separable from the milk
 Dispersed in the milk
 Factors Affecting Composition
65

1. Species
2. Breed
3. Individuality
4. Interval of milking
5. Completeness of milking
6. Frequency of milking
7. Irregularity of milking
8. Day-to-day milking
9. Diseases and abnormal conditions
 Heat Stability of Milk
66

“The heat stability time (HCT) is defined as the time taken

for milk to coagulate while being agitated in a sealed tube
at a set temperature (usually between 120 and 140◦C).”
 Determination of the heat stability (by HCT methods)
showed the milk was least stable in June but most stable
in October
 Milk Mineral and Heat Stability
67

 Calcium and phosphate have been found to play a major

role in the heat stability of milk.
 Addition of calcium salts to milk causes an increase in
serum calcium ions, an increase in calcium in the colloidal
phase, a decrease in pH and a decrease in heat stability.
 Alteration of calcium concentration by the addition of
phosphates or citrates generally causes an
increase in heat stability.
 Milk Protein and Heat Stability
68

 In concentrated milks addition of either whey protein or

purified b-lactoglobulin has a detrimental effect on heat
stability of concentrated milk.
 Addition of casein to milk increased the stability of
concentrates produced from adjusted milk (adjustment of
casein components of milk protein).
 Modification of the casein by hydrolysis causes loss of
heat stability.
69