Effects of heat and high pressure treatment on soluble calcium in milk

Background
Calcium is an essential micronutrient required for biological functions in the body.
Insufficient calcium uptake results in diseases such as rickets in children and osteoporosis in
the elderly. Cow, buffalo, goat, sheep, and camel milks are the main sources providing
dietary calcium to humans around the world.
Calcium in milk exists in two forms with different percentages (% of total calcium):
(1) colloidal calcium phosphate in casein micelle (approx. 66%); (2) soluble calcium in the
soluble milk phase (approx. 34%). Calcium distribution between milk colloidal and soluble
phases has a considerable influence on both the nutritional value of milk (i.e. calcium
bioavailability) and processing of dairy products, such as cheese.
Raw milk needs to be pasteurized or sterilized for safe consumption by humans. Raw
milk can be processed by thermal techniques, such as ultra-high temperature (UHT), and non-
thermal techniques, such as high pressure processing (HPP). Environmental factors and food
processing can modify calcium distribution between milk colloidal and soluble phases, thus
affecting the concentration of soluble calcium in milk.
A trial was carried out to compare the effects of UHT and HPP on soluble calcium
content (% total calcium content) in cow milk. You as a process technologist is asked to write
a report based on the results of this trial.

Materials and method

Raw whole milk from cow, buffalo, and sheep was obtained from a local dairy farm
immediately after milking. Milk samples were transported in cooling boxes to the laboratory
within a few hours of milking, and samples were analyzed on the same day.
Raw whole milk samples were first determined for the total calcium content (mg/100
g milk). In brief, milk was deproteinized by mixing with an equal volume of 20% (w/v)
trichloroacetic acid and holding for 30 min at 4 °C, followed by centrifugation (15,000  g,
30 min, 4 °C) and filtration of the supernatant through filter papers. Total calcium content in
the deproteinized filtrate was determined by atomic absorption spectroscopy (iCE™ 3300
AAS, Thermo Scientific™, USA).
For each type of milk, the analyses were done in 10 replicates, with each replicate
using 100 g of milk to determine the amount (mg) of calcium in it.
UHT and HPP treatments
The raw whole cow milk was further processed using UHT and HPP.
UHT treatment – Aliquots of raw milk (100 g) were filled in glass bottles and heated
at 150 °C for 6 s and immediately cooled down to 20 °C. Milk was first ultra-centrifuged
(100,000  g, 10 min, 20 °C) to obtain the supernatant, and the calcium content in the
supernatant was then determined according to the steps described above.
HPP treatment – Aliquots of raw milk (100 g) were vacuum-packed in polyethylene
bags and processed using a Stansted Iso-Lab 900 High Pressure Food Processor (Stansted
Fluid Power, Stansted, Essex, UK) at 250 MPa and 20 °C for 30 min. Immediately after HPP
treatment, milk was ultra-centrifuged to obtain the supernatant, and the calcium content in the
supernatant was then determined according to the steps described above.
The level of soluble calcium (%) was calculated, by Eq.(1), as the percentage of the
concentration of total calcium in milk after correction for differences in the volume of the
ultracentrifugal supernatant.

Soluble calcium ( mg )
Soluble calcium ( % ) = ×100 % Eq. (1)
Total calcium ( mg )

Data for analysis

Dataset 1
Animal species Milk Replicate Total calcium (mg/100 g)
Cow Raw milk 1 122.55
Cow Raw milk 2 127.56
Cow Raw milk 3 126.26
Cow Raw milk 4 120.11
Cow Raw milk 5 119.69
Cow Raw milk 6 121.12
Cow Raw milk 7 128.47
Cow Raw milk 8 70.68
Cow Raw milk 9 127.87
Cow Raw milk 10 126.64
Sheep Raw milk 1 200.34
Sheep Raw milk 2 206.48
Sheep Raw milk 3 200.44
Sheep Raw milk 4 211.43
Sheep Raw milk 5 214.71
Sheep Raw milk 6 213.62
 Sheep Raw milk 7 206.85
Sheep Raw milk 8 213.30
Sheep Raw milk 9 197.68
Sheep Raw milk 10 207.26
Buffalo Raw milk 1 105.71
Buffalo Raw milk 2 106.21
Buffalo Raw milk 3 106.84
Buffalo Raw milk 4 108.91
Buffalo Raw milk 5 101.35
Buffalo Raw milk 6 104.00
Buffalo Raw milk 7 135.07
Buffalo Raw milk 8 105.08
Buffalo Raw milk 9 101.90
Buffalo Raw milk 10 103.85

Dataset 2
Milk Processing technique Replicate Soluble calcium (%)
Raw cow milk No processing 1 30.15
Raw cow milk No processing 2 29.96
Raw cow milk No processing 3 30.21
Raw cow milk No processing 4 29.51
Raw cow milk No processing 5 32.47
Raw cow milk No processing 6 29.45
Raw cow milk No processing 7 32.18
Raw cow milk No processing 8 30.67
Raw cow milk No processing 9 30.53
Raw cow milk No processing 10 30.48
Raw cow milk UHT 1 20.65
Raw cow milk UHT 2 20.93
Raw cow milk UHT 3 20.26
Raw cow milk UHT 4 20.82
Raw cow milk UHT 5 20.57
Raw cow milk UHT 6 20.85
Raw cow milk UHT 7 20.23
Raw cow milk UHT 8 20.84
Raw cow milk UHT 9 20.65
Raw cow milk UHT 10 20.87
Raw cow milk HPP 1 45.38
Raw cow milk HPP 2 44.78
Raw cow milk HPP 3 46.25
Raw cow milk HPP 4 45.01
Raw cow milk HPP 5 45.55
Raw cow milk HPP 6 44.94
Raw cow milk HPP 7 46.31
Raw cow milk HPP 8 44.96
 Raw cow milk HPP 9 45.37
Raw cow milk HPP 10 44.91

What must be included in the report?

Why do heat and HPP have different effects on soluble calcium content in milk?
What are the mechanisms for heat and HPP effects?
Bonus points
Effects of other food processing techniques on soluble calcium content in milk
Calcium fortification of milk