SEMINAR PRESENTATION

BY:

IDAGU JUDITH EBEHUNG

2014/2015/ND/SLT/FT/003

ON THE TOPIC

DETERMINATION OF CAESIN CONTENT OF DIFFERENT MILK SAMPLES

SUBMITTED IN PARTIAL FULFILLMENT OF THE AWARD OF NATIONAL

DIPLOMA TO THE DEPARTMENT OF SCIENCE LABORATORY TECHNOLOGY

DORBEN POLYTECHNIC BWARI-ABUJA.

16th MAY, 2016

 TABLE OF CONTENTS

CHAPTER 1

Introduction

Background of Study

Objective of Study

Scope of Study

CHAPTER 2

Literature Review

Factors Affecting Milk Composition

Fatty Acids

Residual/Complementary Milk

CHAPTER 3

Methodology

Collection of Samples

Materials

Apparatus

Theory

REFERENCES
 CHAPTER –ONE--

1.0 Introduction

Milk is a complete diet as it contains in its Minerals, Vitamins Proteins,

Carbohydrates, Fats and Water. Average composition of milk from different
sources is given below: Source Water Mineral Protein Fats Carbohydrate of milk
(%) s (%) ns(%) (%) tes (%) Cow 87.1 0.7 3.4 3.9 4.9 Human 87.4 0.2 1.4 4.0 4.9
Goat 87.0 0.7 3.3 4.2 4.8 Sheep 82.6 0.9 5.5 6.5 4.5 Caesin is a major protein
constituent in milk & is a mixed phosphor-protein. Casein has isoelectric pH of
about 4.7 and can be easily separated around this isoelectric pH. It readily
dissolves in dilute acids and alkalis. Casein is present in milk as calcium caseinate
in the form of micelles. These micelles have negative charge and on adding acid to
milk the negative charges are neutralized. Ca2+-Caesinate +
2CH3COOH(aq)^Caesin+(CH3COO)2Ca

Milk is a pale liquid produced by the mammary glands of mammals. It is the

primary source of nutrition for infant mammals before they are able to digest other
types of food. Early-lactation milk contains colostrums, which carries the mother's
antibodies to its young and can reduce the risk of many diseases. It contains many
other nutrients including protein and lactose.

As an agricultural product, milk is extracted from mammals during or soon after

pregnancy. Dairy farms produced about 730 million tones of milk in 2011, from
260 million dairy cows. India is the world's largest producer of milk, and is the
leading exporter of skimmed milk powder, yet has little to no other milk product
exports. The ever increasing rise in domestic demand for dairy products and a large
demand-supply gap could lead to India being a net importer of dairy products in
the future. New Zealand, the European Union's 28 member states, Australia, and
the United States are the world's largest exporters of milk and milk products. China
and Russia were the world's largest importers of milk and milk products. Both
countries were self-sufficient by 2016 contributing to a worldwide glut of milk.
Throughout the world, there are more than six billion consumers of milk and milk
products. Over 750 million people live within dairy farming households.

1.1 AIMS AND OBJECTIVES

The aim of this work is to determine the quantity of casein in different milk
samples

1.1.0 Physical and chemical properties of milk

Milk is an emulsion or colloid of butterfat globules within a water-based fluid that

contains dissolved carbohydrates and protein aggregates with minerals. Because it
is produced as a food source for the young, all of its contents provide benefits for
growth. The principal requirements are energy (lipids, lactose, and protein),
biosynthesis of non-essential amino acids supplied by proteins (essential amino
acids and amino groups), essential fatty acids, vitamins and inorganic elements,
and water.

1.2.0 Lipids

Initially milk fat is secreted in the form of a fat globule surrounded by a

membrane. Each fat globule is composed almost entirely of triacylglycerols and is
surrounded by a membrane consisting of complex lipids such as phospholipids,
along with proteins. These act as emulsifiers which keep the individual globules
from coalescing and protect the contents of these globules from various enzymes in
the fluid portion of the milk. Although 97–98% of lipids are triacylglycrols, small
amounts of di- and monoacylglycerols, free cholesterol and cholesterol esters, free
fatty acids, and phospholipids are also present. Unlike protein and carbohydrates,
fat composition in milk varies widely in the composition due to genetic,
lactational, and nutritional factor difference between different species.

Like composition, fat globules vary in size from less than 0.2 to about 15
micrometers in diameter between different species. Diameter may also vary
between animals within a species and at different times within a milking of a single
animal.
There is a wealth of indirect evidence of very serious possible harms of consuming
dairy foods, and, on the flip side, the evidence that milk prevents fractures is scant.

As we look beyond the headlines, it is hard to think that we should continue to

consume the lactation fluid that exists in nature to nourish and rapidly grow calves.

1.5.0 Nutrition and health

The composition of milk differs widely among species. Factors such as the type of
protein; the proportion of protein, fat, and sugar; the levels of various vitamins and
minerals; and the size of the butterfat globules, and the strength of the curd are
among those that may vary. For example:

 Human milk contains, on average, 1.1% protein, 4.2% fat, 7.0% lactose (a
sugar), and supplies 72 kcal of energy per 100 grams.
 Cow's milk contains, on average, 3.4% protein, 3.6% fat, and 4.6% lactose,
0.7% minerals and supplies 66 kcal of energy per 100 grams. See also
Nutritional value further on

Donkey and horse milk have the lowest fat content, while the milk of seals and
whales may contain more than 50% fat.

1.6.0 Recommended consumption

The U.S. federal government document Dietary Guidelines for Americans, 2010
recommends consumption of three glasses of fat-free or low-fat milk for adults and
children 9 and older (less for younger children) per day. This recommendation is
disputed by some health researchers who call for more study of the issue given
that there are other sources for calcium and vitamin D. The researchers also claim
that the recommendations have been unduly influenced by the American dairy
industry, and that whole milk may be better for health due to its increased ability to
satiate hunger.
1.7.0 Medical research

There is recent evidence suggesting consumption of milk is effective at promoting

muscle growth. Some studies have suggested that conjugated linoleic acid, which
can be found in dairy products, is an effective supplement for reducing body fat.

With regards to the claim of milk promoting stronger bones, there has been no
association between milk consumption and risk of hip fractures in women.[104]

Milk and dairy products have the potential for causing serious infection in newborn
infants. Unpasteurized milk and cheeses can promote the growth of Listeria
bacteria. Listeria monocytogenes can also cause serious infection in an infant and
pregnant woman and can be transmitted to her infant in uterus or after birth. The
infection has the potential of seriously harming or even causing the death of a
preterm infant, an infant of low or very low birth weight, or an infant with an
immune system defect or a congenital defect of the immune system. The presence
of this pathogen can sometimes be determined by the symptoms that appear as a
gastrointestinal illness in the mother. The mother can also acquire infection from
ingesting food that contains other animal products such as, hot dogs, delicatessen
meats, and cheese.

1.8.0 Possible harms

Some studies suggest that milk consumption may increase the risk of suffering
from certain health problems. Cow's milk allergy (CMA) is an immunologically
mediated adverse reaction, rarely fatal, to one or more cow's milk proteins.
Milk from any mammal contains amino acids and microRNA which influence the
drinker's metabolism and growth; this "programming" is beneficial for milk's
natural consumers, namely infants of the same species as the milk producer, but
post-infancy and trans-species milk consumption affects the mTORC1 metabolic
pathway and may promote diseases of civilization such as obesity and diabetes.
Milk contains casein, a substance that breaks down in the human stomach to
produce casomorphin, an opioid peptide. In the early 1990s it was hypothesized
that casomorphin can cause or aggravate autism spectrum disorders, and casein-
free diets are widely promoted. Studies supporting these claims have had
significant flaws, and the data are inadequate to guide autism treatment
recommendations.

A study demonstrated that men who drink a large amount of milk and consume
dairy products were at a slightly increased risk of developing Parkinson's disease;
the effect for women was smaller. The reason behind this is not fully understood,
and it also remains unclear why there is less of a risk for women.[116][117]

Casein

Casein is the name for a family of related phosphoproteins (αS1, αS2, β, κ). These
proteins are commonly found in mammalian milk, making up 80% of the proteins
in cow milk and between 20% and 45% of the proteins in human milk. Casein has
a wide variety of uses, from being a major component of cheese, to use as a food
additive, to a binder for safety matches. As a food source, casein supplies amino
acids, carbohydrates, and the two inorganic elements calcium and phosphorus.

Composition

Casein contains a fairly high number of proline residues, which do not interact.
There are also no disulfide bridges. As a result, it has relatively little tertiary
structure. It is relatively hydrophobic, making it poorly soluble in water. It is found
in milk as a suspension of particles called "casein micelles" which show only
limited resemblance with surfactant-type micellae in a sense that the hydrophilic
parts reside at the surface and they are spherical. However, in sharp contrast to
surfactant micelles, the interior of a casein micelle is highly hydrated. The caseins
in the micelles are held together by calcium ions and hydrophobic interactions.
Any of several molecular models could account for the special conformation of
casein in the micelles.
In un-homogenized cow's milk, the fat globules have an average diameter of two to
four micrometers and with homogenization, average around 0.4 micrometers. The
fat-soluble vitamins A, D, E, and K along with essential fatty acids such as linoleic
and linolenic acid are found within the milk fat portion of the milk.

1.3.0 Proteins

Normal bovine milk contains 30–35 grams of protein per liter of which about 80%
is arranged in casein micelles.

1.3.1 Caseins
The largest structures in the fluid portion of the milk are "casein micelles":
aggregates of several thousand protein molecules with superficial resemblance to a
surfactant micelle, bonded with the help of nanometer-scale particles of calcium
phosphate. Each casein micelle is roughly spherical and about a tenth of a
micrometer across. There are four different types of casein proteins: αs1-, αs2-, β-,
and κ-caseins. Collectively, they make up around 76–86% of the protein in milk,
by weight. Most of the casein proteins are bound into the micelles. There are
several competing theories regarding the precise structure of the micelles, but they
share one important feature: the outermost layer consists of strands of one type of
protein, k-casein, reaching out from the body of the micelle into the surrounding
fluid. These kappa-casein molecules all have a negative electrical charge and
therefore repel each other, keeping the micelles separated under normal conditions
and in a stable colloidal suspension in the water-based surrounding fluid.

Milk contains dozens of other types of proteins beside the caseins including
enzymes. These other proteins are more water-soluble than the caseins and do not
form larger structures. Because the proteins remain suspended in the whey left
behind when the caseins coagulate into curds, they are collectively known as whey
proteins. Whey proteins make up approximately 20% of the protein in milk, by
weight. Lactoglobulin is the most common whey protein by a large margin.
 CHAPTER-TWO

2.0 REVIEW OF LITERATURE

A lot of research work has been carried out on the different aspects of milk
including composition; oxytocin induced changes in milk, mode of action, possible
effects, doses, stage of lactation etc. The most pertinent literature related to the
present study i.e., biochemical profile within bovine mammary secretions is
influenced by an exogenous oxytocin treatment during lactation period has been
reviewed under the following headings:

2.1 Factors affecting milk composition

2.2 Effect of lactation stages on milk composition

2.3 Residual/complementary milk

2.4 Oxytocin

2.5 Oxytocin and milk yield

2.6 Effect of oxytocin on milk composition

2.7 Electrophoretic pattern of protein fractions of milk

2.8 Milk enzymes

2.1 Factors affecting milk composition

The composition of milk is not fixed since many factors influence the end product.
These variations can be related to genetics and environment, level of milk
production, stage of lactation, disease, season, locality and age of the cow
(Gopalkrishan and Lal, 1994). The milk composition also varies within the cows
from milking to milking (Friggens and Rasmussen, 2001). The composition of
milk also differs within species. The lactose content of milk is moderately constant
between dairy breeds, protein varies to some degree but fat varies widely.
For example, it was reported that Guernesy (5.0%) and Jersey (5.5%) breeds
contained the highest milk fat as compared to Holstien (3.5%) that contained the
lowest milk fat percentage (Hurley, 1997). The age of the cow is closely related to
the number of lactations, as an increase in number of lactations is associated with
decrease in fat and solid not fat (SNF) content of milk.

A survey of lactation records of Holstein shows that milk protein contents

decreases 0.1-0.5 units over a period of five lactation or 0.02-0.05 units per
lactation (Rook, 1961). Feed and diet composition are important factors that can
cause changes in milk composition. Protein concentration can be changed to some
extent but lactose scarcely (Sutton, 1989). Although other diseases can affect milk
components level and distribution, mastitis has been the predominate disease
studied. Mastitis results in a reduction of milk fat and increase in milk proteins that
has been attributed to the influx of blood-borne proteins (Shuster, 1991). Mastitis
is also associated with increase in the concentrations of different enzymes in milk
(Auldist et al., 1995).

2.2 Effect of Lactation stages on milk composition

Lactation stage is a major factor affecting the characteristics of milk components

such as fat content, protein, macro-mineral contents, and some of the enzyme
activities but not on the casein: protein ratio or phosphorus content (Schutz et al.,
1990; Decaen and Adda, 1970; Gue!guen, 1971; Benslimane et al.,

2.2.1 Physicochemical composition

The protein content in cow milk rises above the average from 21stweek of
parturition. Increase during the last 10-12 weeks of parturition is rapid. The casein
content of cow varies during the first 60 days and shows an increasing trend after
200 days in lactation. The albumin level of cow milk varies from 0.23% to 0.36%
but increases after 175 days of lactation as influenced by the stage of lactation. The
globulin content also varies from 0.12% to 0.38% (Srivastava, 1993). According to
Coulon et al (1997) milk pH, calcium and the urea content are higher in late
lactation.
Beata (2008) while comparing the daily milk yield, its chemical composition in
milk depending on the lactation that reported that fat and protein content in milk
during test milking at successive lactation stages showed a growing tendency and
the highest concentration of these nutrients was identified between the 200th and
300th lactation day in cow’s milk. Afterwards these contents decreased rapidly
even lower than those at the beginning stage of lactation. Total solids and solids
not fat are also affected by stage of lactation. A study on trend of SNF in Holstein
cows during whole milking cycle indicates the lowest SNF during second month
followed by an increase to the eight month and a rise in nine and ten months of
lactation (Ozrenk and Inci2000). Occurrence of changes in milk content during
lactation; the decrease of milk yield accompanied by an increase in fat and protein
content is the usual description of milk secretion (Kolb, 1987). In contrary, fat and
protein curves had been reported with continuous declined shape and also with the
standard shape as some disagreement has been reported in earlier studies (Schutz et
al., 1990; Stanton et al.1992; Pollott, 2004). Curves for milk and milk components
were normally regarded as independent curves though, these curves are linked, as
milk is a mixture of fat, protein, lactose, vitamins, and minerals, either dissolved or
suspended in water. This connection was previously studied at phenotypic and
genetic level. Milk yield, fat y and protein yield curves follows the standard
lactation curve in 19.3% of lactations, whereas fat and protein percent have
reversed standard shape. High changeability of shapes is contributed by early
phase of lactation with the fat and protein yield curves (Silvestre et al.,2009)

2.2.2 Fatty acids

The fatty acids composition of milk is related to lactation stages. The lactation
stage effect is noticeable and mainly linked to lipid store mobilization in early
lactation, but it only lasts a few weeks each year (Chilliard et al., 2003). Both
lactation stage and

energy balance contribute to variation in milk fat composition and change the
activity of different fatty acid pathways (Stoop et al.,2009).
At initiation of lactation, cows are in negative energy balance, causing
mobilization of adipose fatty acids and incorporation of these long-chain fatty
acids into milk fat (Belyea and Adams, 1990). Concomitantly, de novo synthesis of
short-chain fatty acids by mammary tissue has been inhibited byhigh uptake of
long chain fatty acids (Bauman and Davis, 1974). Eastridge and Palmquist (1988)
studied the pattern of fatty acids in milk fat at 1, 4, 8, and 12 th week of lactation, as
a proportion of those occurring at 16thweek. Two points are striking; firstly, the
proportion of short-chain fatty acids, except for C4:0, is low in early lactation, and
these fatty acids increase, reaching more than 90% of maximal proportions by 8 th
week of lactation. This increase is consistent with the release of inhibition by
adipose mobilization, which is largely completed by 4thto 6 th week of lactation
(Gamsworthy and Huggett, 1992). Second, the synthesis of short-chain fatty acids
is inhibited to different degrees in a pattern that shows increasing inhibition from
C6:0 to C12:0.Lynch et al. (1992) observed the same pattern of change in de novo
synthesis of fatty acids over an entire lactation in cows on a bovine somatotrophin.
An interesting cycling of milk fatty acids composition was observed in cows
injected with recombinant bovine somatotrphin at 14 days intervals. Within a
single injection interval,the percentages of C8:0, Cl0:0, C12:0and C14:0 were
significantly lower at 5th

day post injection than at 12thday post injection, but C18:1 showed the opposite
pattern. It was further observed that the concentration of cholesterol in milk fat
changes with stage of lactation in the same manner as the fatty acids that are
synthesized de novo from acetate (Lynch et al.,1992). Smith et al. (1978) observed
that dietary protected tallow produced fatty acids pattern similar to that of early
lactation. Synthesis of C4.0 is not inhibited at all, which is consistent with its
unique origin from two known pa

thways independent of the inhibitable acetyl-coenzyme A (CoA) carboxylase

pathway (McCart

hy and Smith, 1972).

Similar trends in fatty acids distribution as a percentage of total fatty acids were
shown by other researchers and the variation in actual synthesis of individual
short-chain fatty acids during lactation was relatively small when expressed as
grams of fatty acids per kilogram of milk rather than as proportions (Karijord et al.,
1982; Syrstad, 1982). Effect of lactation stages through thin layer
chromatography was significantly analysed by Armughan and Narayanan (1981)
indicating that with progressive lactation the major changes from cholestrol fat
have been increased in lower fatty acids and declined in oleic acid. The effect on
influence of fatty acids composition has also been studied. Kgwatalala et al. (2009)
reported that Canadian Holstein cows had higher C18; total, lower C10, C12, C14,
and conjugated linoleic acid index, respectively during early lactation compared
with the subsequent lactation stages. Early lactation was characterized by higher
concentrations of oleic acid (C18:1 cis-9), vaccenic acid (C18:1 trans-11), linoleic
acid (C18:2), monounsaturated fatty acids,total polyunsaturated fatty acids, and
lower concentrations of capric acid (C10:0), C10:1,lauric acid (C12:0), C12:1,
myristic acid (C14:0), myristoleic acid (C14:1), palmitic acid (C16:0) and total
saturated fatty acids when compared with the subsequent lactation stages. Neither
the stearoyl-CoA desaturase 1 genotype nor the stage of lactation had an influence
on conjugated linoleic acid concentrations in milk. Oxytocin release, intra-
mammary pressure and milking characteristics in dairy cows has been changed
during lactation were studiedby Mayer et al.(1991). Pre-milking baseline intra-
mammary pressure had its maximum value in early lactation until month 4 and
then decreased to approximately 50% of its initial level. Ejection pressure followed
a similar pattern, but dropped only to approximately 75% of its maximum. This
was due to the constant elevation of pressure increase, reaching its highest level in
late lactation. Time from commencement of stimulation until maximum pressure
exceeded 1 min in

almost all instances even in early lactation and increased throughout lactation.
2.3 Residual /complementary milk

Residual milk under review means the milk which may not be removed from the
udder due to certain reasons. At the start of milking about 15-25 % of the total
amount of milk in the udder is not removed during milking. Residual milk is also
called as complementary milk. Percentage of residual milk after milking as
reported by Schmidt (1971) is 12.8% for machine stripped cows, 14.0% for non
machine stripped cows, 16.8% for cows receiving oxytocin subcutaneously and
12.2% for cows that were measured for one year. Residual milk decreases in
proportion to milk yield as lactation progresses; that is the percentage remains
same throughout the lactation period (Schmidt, 1971). The amount of residual milk
obtained by injecting 10IU of oxytocin intravenously varied roughly as the normal
milk yield within breeds. The fat content of the residual milk was highest at the
peak of lactation and decreased as lactation advanced. The relationship between
quantity and fat test in residual milk was the reverse of that found with normal
milk, i.e., high yields of residual milk were associated with higher fat tests than
were low yields (Swanson and Hinton, 1951).

2.4 Oxytocin

Oxytocin (Greek word: "quick birth") is a nine amino acid peptide (Cys-Tyr-Ile-
Gln-Asn-Cys-Pro-Leu-Gly) with molecular mass of 1007 Daltons. It is synthesized
in hypothalamic neurons and transported down axons of the posterior pituitary
gland into blood to stimulate contraction of myoepithelial cells causing ejection of
milk. The synthetic oxytocin is quite commonly used in human as well as
veterinary medicine.

2.4.1 Mechanism of oxytocin

Milk ejection is an active transport of alveolar fluid into the cisternal compartment.
It consists of contraction of myoepithelial cells that encircle the alveoli like a
basket and transport of the milk through the milkduct system. Tactile stimulation
of the mammary glands through a neuroendocrine reflex arc causes a natural reflex
ecalled milk ejection (Crowley and Armstrong, 1992).
 From the neural lobe of the pituitary gland by the milking stimulus, oxytocin
released into circulation and induced milk discharge during lactation. A cow
having approximately 40 liters of blood would have to release about 0.4 to 2.6IU
of endogenous oxytocin to establish the range of concentrations (Gorwit, 1979;
Sagi etal.,1980). In response to elevated oxytocin blood concentrations, binding to
the oxytocin receptors of the myoepithelial cells causes alveolar contraction
(Soloff et al., 1980). As a result, alveolar milk is forcefully shifted into the
cisternal

space. It causes a rapid increase of pressure within the cistern (Bruckmaier and
Blum, 1996) and an enlargement of the cisternal cavity size (Bruckmaier and
Blum, 1992). Nevertheless, alveolar milk cannot be ejected if milk is not
simultaneously removed from the udder due to the limited cisternal space
(Bruckmaier et al.,1994; Bruckmaierl et al.,1997). Eighty percent of milk stored in
the udder regarded as alveolar fraction, is available after milk ejection that is
stimulated bythe release of oxytocin and myoepithelial contraction. Tactile teat
stimulationcauses milk ejection, either manually or by the milking machine. The
time from the beginning of a tactile stimulation to the occurrence of the milk
ejection lasts from forty seconds upto more than two minutes and increases with
decreasing level of udder filling. That’s why, in end production stages of lactation,
cows need a longer pre-stimulation (Bruckmaier, 2005).

2.4.2 Role of exogenous oxytocin

There are many factors can disturb milk ejection. It has been reported that a
disturbed milk ejection is due to a decreased secretion or complete absence of
oxytocin from the pituitary gland (Bruckmaier, 2005). Oxytoc in release from the
pituitary gland is inhibited in dairy practice during different types of emotional
stresses or for several weeks after parturition in cows. Inhibition of milk ejection is
caused by milking in unfamiliar surroundings which could be eliminated by small
doses of oxytocin injections (Bruckmaier et al., 1993). Anthony et al., (1959) used
oxytocin to aid milk let-down. These differenttechniques do not measure the same
trait.
During early stages of lactation, milk consumption is not an accurate estimate of
the total milk production of a cow (Gifford, 1953). A complete milking by machine
or by hand provides no estimate of milk consumption. Oxytocin treatment did not
have a significant effect on milk consumption or on total milk production
(Schwulst et al., 1966).
 CHAPTER -THREE-

3.0 MATERIALS AND METHOD

250-ml. beaker, filtrations flask, measuring cylinder, glass rod, spatula, china dish,
dropper, and weight bore, different samples of milk, 10% acetic acid.

MATERIALS REQUIRED:

Beakers (250 ml)

Filter-paper

Glass-rod

Weighing machine

Filtration-flask

Funnel

Test-tubes

Burner

Different samples of Milk

1% acetic acid solution

Saturated ammonium sulphate solution.

PROCEDURE:

Take a clean dry beaker, put into it 20mL of sample milk and add 20 ml of
saturated ammonium sulphate solution slowly and with stirring. Fat along with
casein will precipitate out.
Filter the solution and transfer the precipitates in another beaker.

Add about 30 ml of water to the precipitate.

Only casein dissolves in water forming milky solution leaving fat undissolved.

Heat the milky solution to about 40°C and add 1% acetic acid solution drop wise,
when casein gets precipitated.

Filter the ppt., wash with water, and let the ppt. dry.

Weigh the dry solid mass in a previously weighed watch glass.

Repeat the experiment with other samples of milk.

Calcium caseinate + 2CH3COOH(aq) → Casein(s) + (CH3 COO) 2 Ca (aq)

OBSERVATIONS AND CALCULATION:

Volume of milk taken in each case = 20 ml

% of Casein = Weight of Casein / Volume of milk taken × 100

Volume of Milk Taken in each case = 20ml.

Sample No. Quality of Milk Weight of casein percentage of casein

1. PEAK MILK 0.60 3.00%
2 RAM MILK 0.65 3.25%
3. COW MILK 0.85 4.20%
4. GOAT MILK 0.75 3.70

THEORY

Natural milk is an opaque white fluid secreted by the mammary glands of female
mammal.
 The main constituents of natural milk are protein, carbohy-drate, mineral vitamins,
fats and water and are a complete balanced diet. Fresh milk is sweetish in taste.
However, when it is kept for long time at a temperature of 5 degree it become sour
because of bacteria present in air. These bacteria convert lactose

of milk into lactic acid which is sour in taste. In acidic condition casein of milk
starts separating out as a precipitate. When the acidity in milk is sufficient and
temperature is around 36 degree, it forms semi-solid mass, called curd.
REFERENCES

Goff, Douglas (2010). "Homogenization of Milk and Milk Products". Dairy

Science and Technology. University of Guelph. Retrieved February 8, 2011.
"Research Can Lead To Longer Shelf Life For Dairy Products". Sciencedaily.com.
December 23, 2002. Retrieved August 28, 2010.
"Milk contains traces of ash". Chennai, India: Hindu.com. July 10, 2008.
Retrieved August 28, 2010.
"Milk From Cows and Other Animals, web page by Washington Dairy Products
Commission". Havemilk.com. Retrieved August 28, 2010.
Whale. Encarta.
"Milk analysis". North Wales Buffalo. Archived from the original on September
29, 2007. Retrieved August 3, 2009. (Citing McCane, Widdowson, Scherz, Kloos,
International Laboratory Services.)
USDA National Nutrient Database for Standard Reference. Ars.usda.gov.
Retrieved on November 24, 2011. Archived July 7, 2015, at the Wayback
Machine.
Designing Foods: Animal Product Options in the Marketplace. National
Academies Press. 1988. ISBN 978-0-309-03795-2.
"National Nutrient Database for Standard Reference Release 28". United States
Department of Agriculture: Agricultural Research Service.
"Nutrition facts, calories in food, labels, nutritional information and analysis".
NutritionData.com.
"USDA Table of Nutrient Retention Factors, Release 6" (PDF). USDA. USDA.
Dec 2007.
"Nutritional Effects of Food Processing". NutritionData.com.
Jones, Alicia Noelle (2002). "Density of Milk". The Physics Factbook.
Feskanich, D; Willett, WC; Stampfer, MJ; Colditz, GA (1997). "Milk, dietary
calcium, and bone fractures in women: a 12-year prospective study". American
Journal of Public Health 87 (6): 992–7. doi:10.2105/AJPH.87.6.992.
PMC 1380936. PMID 9224182.
Brody T. (1999) "Calcium and phosphate". pp. 761–94 in Nutritional
biochemistry, 2nd ed. Boston: Academic Press, ISBN 0121348369. Heaney,
Robert P.; Weaver, Connie M. (1990). "Calcium absorption from kale". The
American Journal of Clinical Nutrition 51 (4): 656–7. PMID 2321572.
"Calcium and Milk: What's Best for Your Bones and Health?". The Nutrition
Source. Harvard School of Public Health. 2011. Retrieved February 8, 2011.
http://www.weidmann-electrical.com/en/markets-a-products/board/cellulose-
based/laminated-board
 Arthur A. Tracto. Coatings Materials And Surface Coatings. CRC Press. 2006.
pages 19–7 to 19–11
Robert S. Forsyth Waterborne Adhesives for Bottle Labeling
Label Glues
Fankhauser, David B. (2007). "Fankhauser's Cheese Page". Retrieved 2007-09-
23.
Boirie, Y; et al. (Dec 1997). "Slow and fast dietary proteins differently modulate
postprandial protein accretion". Proc Natl Acad Sci U S A 94 (26): 14930–5.
doi:10.1073/pnas.94.26.14930. PMID 9405716.
Field KL, Kimball BA, Mennella JA, Beauchamp GK, Bachmanov AA. (2008).
"Avoidance of hydrolyzed casein by mice". Physiol Behav 28 (93): 189–99.
doi:10.1016/j.physbeh.2007.08.010. PMC 2254509. PMID 17900635.
Louis Malcmacher. "Enamel Remineralization: The Medical Model of Practicing
Dentistry". Dentistry Today.
Glenn Walker, Fan Cai, Peiyan Shen, Coralie Reynolds, Brent Ward, Christopher
Fone, Shuji Honda, Megumi Koganei, Munehiro Oda and Eric Reynolds (2006).
"Increased remineralization of tooth enamel by milk containing added casein
phosphopeptide-amorphous calcium phosphate". Journal of Dairy Research 73
(1): 74–78. doi:10.1017/S0022029905001482. PMID 16433964.
Campbell, T.C. and Campbell, T.M. 2006. The China Study. Benbella Books
Dr. Neal Barnard 2004, Breaking the Food Seduction, St Martin's Griffin
Millward C, Ferriter M, Calver S, Connell-Jones G. Gluten- and casein-free diets
for autistic spectrum disorder" Cochrane Database of Systematic Reviews 2008,
Issue 2. Art. No.: CD003498. doi:10.1002/14651858.CD003498.pub3.
Truswell, A.S. (2005), "The A2 milk case: a critical review", European Journal of
Clinical Nutrition 59: 623–631, doi:10.1038/sj.ejcn.1602104, PMID 15867940,
retrieved 29 June 2014
Truswell, AS (2006). "Reply: The A2 milk case: a critical review". European
Journal of Clinical Nutrition 60: 924–925. doi:10.1038/sj.ejcn.1602454. Retrieved
30 July 2014.
1 February 2009, EFSA review of scientific literature on A1 and A2 milk, Review
of the potential health impact of β-casomorphins and related peptides
Solinas, C; et al. (Oct 2010). "Cow's milk protein allergy". J Matern Fetal
Neonatal Med. 23 (Suppl 3): 76–9. doi:10.3109/14767058.2010.512103.
PMID 20836734.