Journal Pre-proof

Moderate electric field pasteurization of milk in a continuous flow

unit: Effects of process parameter, energy consumption, and shelf-
life determination

Asaad Rehman Al-Hilphy, Alaa Riyadh Abdulstar, Mohsen

Gavahian

PII: S1466-8564(20)30514-2
DOI: https://doi.org/10.1016/j.ifset.2020.102568
Reference: INNFOO 102568

To appear in: Innovative Food Science and Emerging Technologies

Received date: 28 June 2020

Revised date: 1 October 2020
Accepted date: 17 November 2020

Please cite this article as: A.R. Al-Hilphy, A.R. Abdulstar and M. Gavahian, Moderate
electric field pasteurization of milk in a continuous flow unit: Effects of process parameter,
energy consumption, and shelf-life determination, Innovative Food Science and Emerging
Technologies (2020), https://doi.org/10.1016/j.ifset.2020.102568

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Moderate electric field pasteurization of milk in a continuous flow unit: effects

of process parameter, energy consumption, and shelf-life determination

Asaad Rehman Al-Hilphy1. Alaa Riyadh Abdulstar1, Mohsen Gavahian2,*

1
Department of food science, College of Agriculture, University of Basrah, Iraq

2
Department of Food Science, National Pingtung University of Science and

of
Technology, Pingtung 912, Taiwan, ROC

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* Corresponding author:

Dr. Mohsen Gavahian

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National Pingtung University of Science and Technology, Pingtung 912, Taiwan
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Tel: +886-8-7703202; Email: mohsengavahian@yahoo.com

ORCID: 0000-0002-4904-0519
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Running head title: Moderate electric field pasteurization of milk

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Abstract

This study presents a new non-thermal moderate electric field (MEF) process for

milk pasteurization. To evaluate the applicability of this process, fresh milk was

treated by MEF at electrical field strength (EFS) of 8.33, 14.58, and 20.80 V/cm and

mass flow rates (MFR) of 0.018, 0.042 and 0.077 kg/s to compare the

microbiological quality, alkaline phosphatase activity, chemical composition, and

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some physical properties of the product with those of conventionally pasteurized

(thermal pasteurization product) and raw milk (RM). The changes in the total count of

bacteria (TCB) and titratable acidity of samples were observed during 18 days of

storage at 5˚C for shelf-life estimation. The results showed that MEF reduced energy

consumption by 63% as compared to thermal pasteurization. Also, MEF treatment

inactivated coliforms (100%), Staphylococcus aureus (100%), psychrophiles (100%),

yeasts and molds (100%), total count Bactria (116%) alkaline phosphatase (100%)

compared to RM, while keeping the processing temperature below 22˚C. Also, TCB

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of sample was reduced by increasing the EFS from 8.33-20.8 V/cm and decreasing

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the MFR from 0.0774 - 0.0185 kg/s. The longest shelf-life of MEF samples, i.e., 15
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days, was observed at EFS of 20.80 V/cm and MFR of 0.018 kg/s which was better
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than that of thermal pasteurization samples, i.e., 9 days.
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Keywords: emerging technologies; milk; moderate electrical field; energy

consumption; pasteurization.
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Abbreviations
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A section area (m2)

AP alkaline phosphatase

EFS, electrical field strength (V/cm)

FP freezing point (˚C)

I current (A)

L distance between electrodes (cm)

L pipe length (m)

MEF moderate electric field

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MFR mass flow rate of milk (kg/s)

Nj final bacterial count (CFU/mL)

Noj initial bacterial count (CFU/mL)

Pcons. consumed power (kWh)

R1 outer pipe with a large radius (cm)

R2 inner pipe with a small radius (cm)

RM raw milk

Re Reynolds number

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t residence time (s)

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t2-t1 processing time (min) -p
TA titratable acidity (%)
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TCB total count of bacteria (CFU/mL)

TM temperature of milk
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V voltage (V)
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1. Introduction
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Milk is regarded as a wholesome food due to its balanced content of protein, fat,

lactose, minerals, and vitamins (McAuley et al., 2016). However, fresh milk usually

needs to be sterilized to improve its microbiological safety and shelf-life. The

conventional methods of sterilization rely on thermal treatments at various time-

temperature combinations, ranging from low-temperature long-time pasteurization

(e.g., about 63 °C for 30 min) to ultra-high-temperature (UHT) sterilization (e.g.,

about 135-154 °C for a couple of seconds). Regardless of the time-temperature

program, heat affects the milk chemistry and reduces the nutritional,
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physicochemical, and sensory qualities of the product (Prasantha &Wimalasiri,

2019).

On the other hand, food consumers are nowadays seeking for safe, healthy, and

minimally processed food with a “fresh-like” taste. Such a market demand initiated

an intense competition in the food industry to develop alternative processing

technologies that can ensure safety while keeping the “fresh-like” characteristics of

the foods (e.g., pasteurized milk). In this regard, researchers explored the

applicability of emerging food processing technologies, specifically the non-thermal

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processes such as high-pressure processing, pulsed electric fields, and ultraviolet

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(Nunes &Tavares, 2019). These attempts resulted in the emergence of several novel
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decontamination technologies that are designed based on non-thermal technologies.
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Besides, previously conducted research showed that electricity poses both thermal,

that is, ohmic heating, and non-thermal effects on the microorganisms (Gavahian et
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al., 2012; Gavahian &Tiwari, 2020). The latter process regarded as a moderate
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electric field (MEF) that has been used for extraction, blanching, and microbial

inactivation (e.g., Escherichia coli) (Gavahian et al., 2018; Machado et al., 2010).
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Regarding the application of electricity for treatment of milk, a good effort was made
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in a preliminarily study to limit the final temperature of the milk pasteurization to 40°C

when a laboratory-scale ohmic system was used at a relatively high voltage of 80-

220 V to take advantage of both thermal and non-thermal effects of the electricity.

However, MEF has good potential to be used as a decontamination technology at

low temperatures, that is, non-thermal sterilization (Li et al., 2020). Despite such

good potential, the best of our knowledge, the applicability of MEF as an innovative

non-thermal technology for milk pasteurization at a pilot-scale has not been

documented in the literature. Therefore, the present study aims to develop a new
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non-thermal MEF process at a pilot-scale for extending the shelf-life and

microbiological safety of milk, to assess the engineering aspects (e.g., energy

consumption and productivity) of this new process in comparison with those of the

conventional thermal pasteurization, and to study some quality characteristics of milk

(e.g., chemical composition) as affected by MEF and conventional thermal

pasteurization.

2. Materials and methods

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2.1. Materials

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Fresh whole bovine milk, as the raw milk (RM), was supported by the Agricultural

Research Station of College of Agriculture of the University of Basrah and was kept
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at 4˚C before experiments. Stainless steel 304 and Teflon (with the density of 2200
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kg/m3) were purchased from Prior Plastic (Shenzhen, China) and Calico Metal
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(Mumbai, India), respectively. All the chemicals used in the present study were of

analytical grades and purchased from Sigma-Aldrich (St. Louis, United States)
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unless other suppliers are specified.

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2.2 Moderate electric field (MEF) milk pasteurizer set up

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A new MEF system for milk pasteurization was designed, manufactured, and tested

in the Food Engineering Laboratory of the Department of Food Science at the

College of Agriculture, University of Basrah (Figure 1). This system consists of three

main units, namely a non-thermal pasteurization unit, a pumping unit, and a power

supply and process monitoring unit.

As indicated in Figure 1a, the non-thermal pasteurization unit includes a pipe (2) as

an outer electrode made of stainless steel type 316 with a length of 132 cm, inner

electrode (pipe 1) with an inner diameter of 3.9 cm, and a pipe (1) made of stainless
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steel type 316 with a length of 150 cm and an outer diameter of 1.8 cm. These

components were fixed via a Teflon ring (4) to isolate these two tubes. These tubes

act as the MEF electrodes with a distance i.e., the electrode gap, of 1.05 cm (Figure

1b). The pasteurization unit was isolated by a plastic tape wrapped on the outer tube

with a thickness of 3 mm, and this unit was supported with a plastic tube with a

diameter of 1.25 cm (6) ending with a valve (8) to control the exit of the pasteurized

milk from it (32). A thermocouple (7) and a ground conduction electrode (9) were

also installed in this section. The inner tube (1) is connected to the outer tube (2) via

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a plastic tube (5) 1.25-cm diameter which transfers the milk passing through the

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inner tube into the outer tube so that the milk can be located between the two tubes,
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i.e, electrode gaps, and be treated by MEF.
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The pumping unit consists of a 20-L tank (16) made of stainless steel 316 and a 70
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W pump (PSB-02B, Zhejiang, China) with the discharge capacity of 20 L/min. Also,

plastic tubes with a diameter of 1.25 cm were installed to transfer milk to the inner
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tube (1), that is, the non-thermal pasteurization unit. The pumping unit was equipped
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with a pressure gauge (11) to measure the milk pressure in the system. The third

unit of this system, i.e., the power supply and process monitoring unit, consists of a
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source of alternating electric current (AC) (20) equipped with a voltage variac

(TDCG2, Xinjiang, China) (19) with the range of 0-220 V, electric wires (18), and a

power switch (28). It also includes a voltage meter (30), an electrical current meter

(27), and a digital temperature gauge (LTR5/LEA Electronic, Italy) to monitor the

process voltage, electrical current, and milk, respectively. The MEF system was

equipped with two thermocouples; one (6) on the milk exit tube (7) and another one

(23) on the milk inlet tube (12).

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2.3. MEF pasteurization process

RM was pumped (14) from the tank (16) to the non-thermal pasteurization unit at

three mass flow rates (MFR) of 0.0185, 0.0425, and 0.0774 kg/s. The MFR was

calculated by measuring the weight of the product (kg) at a unit of time (s) with three

replications for each treatment. When the milk traveled through the plastic tube (5)

into the tube (2), the milk fills the space between the two tubes (system electrodes)

and acted as an electric resistance. The power supply applied voltages of 10, 17.5,

and 25 V through these electrodes to treat the milk at various electrical field

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strengths (EFS). After MEF treatment, the product was collected in a storage tank

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and refrigerated at 5˚C before further experiments.
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2.4. Conventional pasteurization process
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Milk was pasteurized at 72˚C for 15-s in a batch system. In this regard, 1 L of RM
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put in a glass container and heated in a 2 kW water bath (GLF 1012, Germany) at a

temperature of 76˚C. This heating step was continued with a holding time of 15
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seconds at 72˚C. The pasteurized milk was then cooled down using tap water and

refrigerated at 5˚C before further experiments.

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2.5. Process evaluation and engineering calculations

2.5.1. Reynolds number

Reynolds number (Re) of the milk in the MEF system was calculated according to

Eq. (1).

(1)

Where, is Reynolds number (dimensionless), is the mass flow rate (kg/s),

is the dynamic viscosity (Pa/s) and is the equivalent diameter (m).

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2.5.2. Electric field strength (EFS)

EFS at voltages of 10, 17.5, and 25 V was calculated according to Eq. (2) (Gavahian

et al., 2019).

(2)

Where, EFS, V, L are the electric field strength (V/cm), voltage (V), and the electrode

gap (cm), respectively.

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As the electrodes used in this device were cylindrical, the EFS was calculated

according to Eq. (3).

) )
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[ ]
;
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Where, R1 is the inner tube with a small radius (cm), R2 is the outer tube with a
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bigger radius (cm).

2.5.3. Residence time

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The residence time of milk between the electrodes during MEF treatment was
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calculated according to Eq. (4) (Maroulis &Saravacos, 2003).

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(4)

Where, t is residence time (s); A is cross-sectional area (m2); Ld: is tube length (m);

and MFR is mass flow rate (kg/s).

2.5.4. Productivity

The productivity of the system was calculated by measuring the volume of the MEF-

treated milk as it left the system per unit of time and reported in liter per hour (L/h).

2.5.5. Consumed power and specific energy consumption

Consumption of electric power (kWh) was calculated according to Eq. 5.

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(5)

Where, and t are the consumed power (kWh) and treatment time (h),

respectively.

Besides, the specific energy consumption (SEC) during the treatment of milk

calculated according to Eq. (6) (Raso et al., 2016):

∫ ) ) (6)

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Where, is the specific energy consumption (kJ/kg), is the voltage (V), is the

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time (s), and is the mass of the treated milk.
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2.6. Evaluation of physical and chemical properties of milk
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The Ostwald glass viscometer (size A) (PSL-Rheotek, USA) was used to determine

the viscosity of milk samples at 25 °C according to (AOAC, 2016). The titratable

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acidity (TA), and pH, were performed according to the Official Methods of Analysis
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(AOAC, 2016). The milk density and the percentage of moisture, protein, fat, ash,

and lactose were measured using a Lacto flash apparatus (Funke Gerber, German).
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These tests were performed in triplicates for all the MEF, thermal pasteurization, and
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RM samples.

2.7. Biological investigations

Microbiological tests were performed on RM, thermal pasteurization, and MEF

samples at days 0, 3, 6, 9, 12, 15, and 18 of refrigeration. Microbiological assays

were performed in a similar method that is described in a previous study (Keyser et

al., 2008). Briefly, 11 mL of milk sample (either RM, MEF, or thermal pasteurization)

was added to 99 mL of peptone water, which contains 0.1% peptone, under sterile

conditions. The sample was mixed well, and then the decimal dilutes were prepared
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to estimate the total count of bacteria (TCB), psychrophiles, coliforms, S. aureus,

yeasts, and molds as well as the activity of alkaline phosphatase.

To prepare a culture medium for TCB calculation, 28 g of nutrient Agar (Himedia,

India) dissolved in 1 L of distilled and sterilized water. After sample inoculation,

incubation was performed at a temperature of 37 °C for 48 h. Finally, TCB was

calculated considering the sample dilution and the number of colonies in the culture

medium.

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For determination of psychrophiles, the culture medium was prepared in the same

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method as the TCB assay. However, the incubation temperature of 5 °C for 48 h was

used to measure the number of psychrophiles.

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For determination of coliforms, An aliquot of 51.5 g of MacConkey agar (Himedia,

India) dissolved in 1 L of distilled and sterilized water to prepare the culture medium.
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After inoculation, the media were incubated at a temperature of 37 °C for 48 h to

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calculate the number of coliforms.

For determination of the S. aureus, an aliquot of 111 g of mannitol salt agar

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(Himedia, India) dissolved in one liter of distilled water. Then the culture medium was
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then sterilized at 121 °C for 15 min. After sample inoculation, incubation was

performed at a temperature of 37 °C for 48 h to determine the S. aureus count.

To assess the total number of yeasts and molds, an aliquot of 51.5 g of malt extract

agar (Himedia, India) dissolved in 1 L of distilled and sterilized water Then, serially

diluted samples were added, and the mixtures were incubated at a temperature of

25°C for a period of 48 h. Afterward, the growing yeasts and molds were calculated.
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The presence of the alkaline phosphatase enzyme in milk samples was verified

using a commercial test kit (Egyptian Company for Biotechnology, Egypt) according

to the instruction provided by the supplier.

Decimal reduction time (D-value), that is, the time required in minutes for the

elimination of 90% of the microbial population, was calculated for TCB according to

Eq. (7) (Singh &Heldman, 2014).

(7)

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Where, Noj is the initial (before pasteurization) TCB (CFU/mL), Nj is the final TCB

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(after pasteurization) (CFU/mL), and t2-t1 is the processing time (min).
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2.8. Sample evaluation during storage and shelf-life determination
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Samples were packaged under hygienic conditions in 10 mL airtight polyethylene
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terephthalate tube. The samples were then kept at 5˚C for 18 days. Periodic checks

(every 72 hours) were carried out during the storage. These include TA, pH, and
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microbial tests (as described in section 2.7. of the present study). Besides, as
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defined by the FDA, the total bacteria count of pasteurized milk should not exceed

100,000 per mL (FDA, 2017). This value was considered as an indicator for the
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microbiological shelf-life of samples to compare their stability during the refrigerated

storage (Juffs &Deeth, 2007) (Al-Hilphy, 2013) .

2.9. Statistical analysis

Statistical analysis of experimental data was performed by version 21 SPSS using a

complete randomized design. Also, the least significant difference test was used to

compare means at the level of 0.05. All the experiments in the present study were

conducted in triplicates and the results reported as mean ± standard deviation.

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3. Results and discussion

3.1. Processing parameters

3.1.1. Reynolds number

Figure 2 illustrates that, regardless of EFS values, the Re of samples during the

process was increased as the MFR increased. This is because the MFR is directly

proportional to Re according to Eq. (1). In the present study, the milk in the MEF

system showed a laminar flow with the minimum and maximum Re of 152.31 and

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648.50, respectively (Singh &Heldman, 2014).

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3.1.2. Productivity and residence time

According to Figure 3, the productivity of the MEF process in the present study was
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in the range of 66.6-278.6 L/h. The data showed that productivity increased
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significantly (P<0.05) with increasing MFR at all values of the EFS. This increase is
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due to an increase in the discharge of treated milk by MEF at higher MFR. Also, the

residence time of milk in the MEF was in the range of 17.8-74.7 seconds and
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decreased significantly (P <0.05) with the increase of the MFR at all EFS values.
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This observation is also related to the increase in the speed of milk at a higher MFR.
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3.1.3. Consumed power and specific energy consumption

The electrical current in the MEF process ranged between 18.4 and 32.4 A.

According to Figure 4, electrical current was increased with increasing EFS and with

decreasing MFR. This variation in electrical current affects the consumed power in

MEF at various process conditions, i.e., different EFS and MFR. As a result,

electrical power consumption in the MEF system ranged between 1.10 and 4.86 kWh

which was less than that of thermal pasteurization, i.e., 12 kWh. It means that

replacing the traditional thermal pasteurization system with the MEF system can
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save a substantial amount of energy required for milk pasteurization. This

observation was in line with that reported in the literature (Abdulstar et al., 2020).

Furthermore, Figure 4 shows that the SEC was decreased as MFR increased. An

increase in MFR reduces the residence time of milk under MEF conditions and

decreased electrical current passed through the sample which resulted in a decrease

in SEC by increasing MFR. Moreover, it was observed that the SEC increased as

EFS increased due to the application of higher electrical energy. The highest SEC

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was 2.43 kJ/kg at EFS of 20.80 V/cm and MFR of 0.0185 kg/s while the lowest SEC

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was 0.55 kJ/kg at EFS of 8.33 V/cm and MFR of 0.0774 kg/s. SEC of MEF system

was superior to that of the thermal pasteurization system which yielded a relatively
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high SEC of 3.60 kJ/kg.
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3.2. Effects of the process on physical and chemical properties of milk
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3.4.1. Product temperature

According to Table 1, the temperature increase of the sample during MEF was in the
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range of 13.0-15.8˚C. The highest increase in the MEF-treated temperature was

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observed when EFS was 20.80 V/cm and MFR was 0.0185 kg/s while MEF at EFS
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of 8.33 V/cm and MFR of 0.0774 kg/s increased the temperature of milk from 7.0 to

20.0˚C. This process conditions limited the changes in milk temperature to 13˚C,

which was the lowest temperature change among all the treatments. However,

thermal pasteurization treatment increased the sample temperature by about 65˚C to

achieve a similar goal, i.e., the pasteurization of milk. As can be seen from the data,

the MEF-based system designed in the present study minimized the changes in the

milk temperature which can minimize the undesirable heat-induced changes in the

milk. In a preliminary effort of developing a MEF system for milk pasteurization, mild

ohmic heating was applied to milk which yielded a product with a final temperature of
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40˚C (Al-Hilphy, 2013). The final temperature obtained in the present study was

much lower than that of the above-mentioned preliminary work.

3.4.2. Milk density

The densities of MEF-treated milk samples were in the range of 1025 to 1031 kg/m3.

According to Table 1, the density RM milk was not changed significantly by altering

the MFR when the EFS was 8.33 V/cm. However, at higher EFS (14.58 and 20.80

V/cm), the milk density decreased significantly (P<0.05) as MFR increased. Also,

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increasing the EFS at the same MFR decreased milk density. Similarly, the density

of the thermal pasteurization product was different from that of RM. Such

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observation might be attributed to the slight decrease in the percentage of protein or
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protein participation on the electrodes which may affect the milk density (Ayadi et al.,
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2004; Cappato et al., 2017; Prasantha &Wimalasiri, 2019).
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3.4.3. Milk viscosity

Table 1 showed that EFS and the MFR did not affect the viscosity of the milk.
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Samples Pasteurized by MEF had a viscosity in the range of 0.001803-0.001855

Pa.s which was similar to that of thermal pasteurization, that is, 0.00186 Pa.s. This
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was in agreement with a previous study that mentioned that conventional thermal
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treatment had no significant effect on milk viscosity (Prasantha &Wimalasiri, 2019).

3.4.4. Milk freezing point

The results in Table 1 showed that the milk freezing point ranged between -0.53 to -

0.55˚C and was not affected by process parameters. These results suggested that

the freezing point of milk samples can comply with those recommended by

regulatory bodies (Cais-Sokolińska et al., 2018).

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3.4.5. Chemical composition

According to Table 2, there were no significant differences (P>0.05) between the

chemical composition of samples, that is, MEF did not alter the chemical composition

of the milk. This observation was consistent with a previous paper which indicated

that the mild heat treatment has no significant effect on the chemical composition of

milk (Manzoor et al., 2020). The pH of the milk was unchanged after MEF and

thermal pasteurization processes which is in line with a previous observation on the

effects of alternating electrical currents on the pH of milk (Altemimi et al., 2018).

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Although no significant variations in the composition of macro-elements were

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observed, future studies may further assess the amino acids, lipid profile, and the
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vitamins of the samples in more detail to provide nutritional information about MEF-
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treated milk.
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3.3. Decontamination effects of the process

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3.3.1. Effects of MEF on the microbial count and alkaline phosphatase activity

It was previously hypothesis that MEF can cause cell inactivation through
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electroporation and electro-permeabilization, that is, the formation of pores in the cell
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membranes, destabilization of the cell membranes bilayer, changes in cell

membrane permeability, membrane swell and destruction (Gavahian et al., 2018;

Gavahian &Tiwari, 2020). The observation in the present study supports this

hypothesis as MEF caused a significant decrease in the microbial population at

relatively low temperatures (Table 3). According to the results, the TCB was

decreased significantly (P<0.05) as the EFS increased and the MFR decreased. It

was previously explained that longer exposure times to the MEF increase the

severity of electroporation and cell inactivation (Mok et al., 2019). Similar

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observations in terms of the effects of EFS on microbial inactivation during MEF

processing were reported in the literature (Machado et al., 2010). The decrease in

the MFR means an increase in residence time of the milk in the MEF, that is, a more

intense non-thermal process due to an increase in the exposure of milk to the

electric field. For example, the reduction in TBC was 84% and 92% when MFR was

0.0774, and 0.0185 kg/s, respectively. Furthermore, the results suggested that MEF-

treated milk can comply with the microbiological criteria of pasteurized milk as

recommended by Food and Drug Administration (FDA) because the TBC of the

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MEF-treated samples was below 100,000 per mL (5 log CFU/mL) (FDA, 2017). The

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results also showed that the level of coliforms, S. aureus, psychrophiles, yeasts and
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molds in MEF-treated samples were non-detectable.
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The results also showed the alkaline phosphatase, which was present in the RM
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sample, was inactivated by both MEF and thermal pasteurization processes (100%).

Alkaline phosphatase is considered a milk pasteurization indicator, that is, inactivate

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this enzyme to undetectable level is a sign of complete pasteurization (Suebsiri et

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al., 2019). The inactivation of this enzyme reported in the literature when the

researcher explored emerging thermal (ohmic heating) and non-thermal

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pasteurization approaches (Shamsi et al., 2008; Suebsiri et al., 2019).

3.3.2. D-value

The results showed that D-value was decreased significantly (P<0.05) as the EFS

increased (Figure 5). For example, increasing EFS from 8.33 to 20.80 V/cm reduced

D-value from 1.47 to 0.70 min at the same MFR of 0.0425 kg/s. Furthermore,

increasing MFR reduced D-value due to a reduction in MEF treatment. The greatest

D-value was 2.55 min which was observed at EFS and MFR of 20.80 V/cm and
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0.0185 kg/s, respectively. This observation was in line with those reported by

(Machado et al., 2010) and (Manzoor et al., 2020).

3.4. Quality parameters of milk during storage

3.6.1. The effect of storage time on titratable acidity (TA)

It was observed that the TA increased with increasing storage period, but the rate of

this increase was different for different samples (Figure 6). The highest and lowest

rate of TA changes were observed for RM and a MEF-treated sample (EFS of 20.8

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V/cm and MFR of 0.0185 kg/s), respectively. Also, the rates of TA increase in all the

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MEF samples were less than that of the thermal pasteurization sample. The change

in TA is attributed to microbial activity during storage as described in section 3.5 of

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the present study.
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3.6.2. The effect of storage time on pH

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Figure 7 shows that the pH of the RM and thermal pasteurization sample decreased

significantly after 3 and 18 days of storage while the pH values of the MEF samples
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were stable during the storage period. Changes in the milk pH during storage are
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related to the microbiological activity (McAuley et al., 2016). Therefore, an effective

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decontamination approach, such as MEF, can limits the changes in the pH of milk

during storage.

3.6.3 The effect of storage time on total bacterial count

According to Figure 8, the TCB of all the samples increased during storage. At the

end of the storage period, pasteurized milk at EFS of 20.8 V/cm and MFR of 0.0185

kg/s had the lowest TCB, that is, 51200 CFU/mL. On the other hand, at the end of

the storage period, RM and thermal pasteurization samples had TCB of 160500

CFU/mL and 109400 CFU/mL, respectively.

 Journal Pre-proof

3.7. Shelf-life of milk samples

The results showed that the EFS of 20.80 V/cm poses the longest shelf-life, that is,

15 days as its TCB reached 334, 402, 464 X10 2 CFU/mL for MFR of 0.0185, 0.0425

and 0.0774 kg/s, respectively. On the other hand, the thermal pasteurization sample

had a shelf-life of 9 days and its TCB reached 452 x10 CFU/mL. It was previously

reported that a shelf-life of 10 to 15 days is expected for pasteurized milk at

refrigerator conditions (Juffs &Deeth, 2007).

of
Conclusion

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The findings showed that it is possible to produce a “fresh-like” pasteurized milk by

the MEF process developed in the present study. MEF was found to be an efficient
-p
non-thermal pasteurization technique as it yielded a product with a shelf-life of 15
re
days by reducing the TBC, and inactivating coliforms, S. aureus, psychrophiles,
lP

yeasts and molds, and alkaline phosphatase. Also, MEF was found to be an energy-

saving process and higher energy efficiency of the system can be expected at higher
na

EFS and lower MFR. Furthermore, to achieve a pasteurized milk with a lower TCB,
ur

high values of EFS and low values of EFS of MEF can be recommended.
Jo

CRediT author statement

A-R. A-H.: Conceptualization and aims, Writing - Original Draft, Software, Supervision

Methodology, Resources and Data Curation; A-R. A: Investigation, Validation. Formal

Analysis, Methodology; M.G.: Writing - Review & Editing, Supervision, Visualization.

Declaration of interests

☒ The authors declare that they have no known competing financial interests or personal
relationships that could have appeared to influence the work reported in this paper.
 Journal Pre-proof

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(a)
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(b)

Figure 1. A schematic representation of the moderate electric field (MEF) milk pasteurizer
(a); and the design of the electrodes of MEF (b). 1: Stainless steel inner tube; 2: Stainless
steel outer tube; 3: Insulator; 4: Teflon ring 5: Plastic tube; 6: Pasteurized milk output tube;
7, 23: Thermocouples; 9: Tube; 11: Pressure gauge; 12, 17, 18: Electrical wires; 13, 24:
Thermocouple wires; 14: Pump; 15: Milk; 16: Storage tank; 19: Voltage variac; 20: Source of
alternating current electricity; 21, 22: Temperature gauges; 25: System body structure; 26:
Tire; 27: Gauge for current; 28, 29: Operation switches; 30: Voltage gauge.
 Journal Pre-proof

800

700
8.33 V/cm
a
600 a
14.58 V/cm
a
500 20.8 V/cm
Re

400
b
b
300 b

200
c
c
100 c

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0
0 0.02 0.04 0.06 0.08 0.1

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Mass flow rate (kg/s)
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Figure 2. Reynolds number (Re) as affected by mass flow rate and electric field strengths. Different
letters refer to significant differences at level 0.05, errors bars represent the standard error.
re
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350 90
80
300
A
c 70
250

Residance time (s)

Productivity (L/h) 60
200 50
Productivity (l/h)
150 b 40
Residance time (s)
B 30
100
20
a C
50 productivity of
thermal 10
0 pasteurization= 0
0 0.02 0.04 0.06 [Y VALUE]
0.08 l/h 0.1
Mass flow rate (kg/s)

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Figure 3. Productivity and residence time as affected by mass flow rate. Different letters

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refer to significant differences at level 0.05, errors bars represent the standard error.
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Figure 4. Consumed power and specific energy consumption (SEC) versus mass flow rate at
different electrical field strengths. TP: thermal pasteuriation. Different letters refer to
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significant differences at level 0.05, errors bars represent the standard error.
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3
0.0185 kg/s
a
2.5 0.0425 kg/s
0.0774 kg/s
2 b
D-value (min)

1.5
a c
1 b
a c
b
0.5 c

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0
5 10 15 20 25

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Electric field strenth (V/cm)
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Figure 5. Decimal reduction time (D-value) of the total count of bacteria as influenced by
electric field strength and mass flow rates. Different letters refer to significant differences at
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level 0.05, errors bars represent the standard error.
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0.7
8.33 V/cm 14.58 V/cm 20.8 V/cm TP RM
a
0.6

Titratable acidity (%)

0.5 b

c
0.4
d a
0.3 e b
f b b a a
g d c
e e e
0.2 g g f f c
c
g
g f a
d d b
0.1 e d c
g f

0
0 3 6 9 12 15 18
Storage period (day)
a

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0.7
8.33 V/cm 14.58 V/cm 20.8 V/cm TP RM
0.6

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Titratable acidity (%)

b a
0.5
c
0.4

0.3 e
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d
c c
b b a a
g f d b a
g e d
0.2 g e e d c
f f
re
a
g e
0.1 g ff d c b

0
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0 3 6 9 12 15 18

b Storage period (day)

8.33 V/cm 14.58 V/cm 20.8 V/cm TP RM

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0.7
a
0.6
Titratable acidity (%)

0.5 b
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c
0.4
d b b a
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0.3 e c c
g f d d b
e
0.2 g g f e e
d c
f
b a
0.1 g e d c a
f
0
0 3 6 9 12 15 18
Storage period (day)

c
Figure 6. Development of titratable acidity of MEF pasteurized milk at different electric field
strength and mass flow rates (a: 0.0185 kg/s; b: 0.0425 kg/s and c: 0.0774 kg/s) as compared
with those of raw milk (RM) and thermal pasteurization (TP) during storage, Different letters
refer to significant differences at level 0.05, errors bars represent the standard error .
 Journal Pre-proof

8 8.33 V/cm 14.58 V/cm 20.8 V/cm TP RM

7 a b b b c c d d e e f f g g

aa c c dc e
6 b e f
a b f g g
a c
5 d
e
pH
4
f
3
g
2

0
0 3 6 9 12 15 18

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Storage period (day)
a

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8
8.33 V/cm 14.58 V/cm 20.8 V/cm TP RM
c c d
7 a b d e e f f g g
a a b
c d
6
a a b
b b
c
-p d e
e
f
f g g
c
5 d
e
pH

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4 f
3 g
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0
0 3 6 9 12 15 18
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Storage period (day)

b
8 8.33 V/cm 14.58 V/cm 20.8 V/cm TP RM
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a b b c d e f
7 a c d e f g g
aa bb c c d d f
6 a b e e f g g
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c
5 d
e
pH

4 f
g
3

0
0 3 6 9 12 15 18
Storage period (day)
c
Figure 7. Development of pH of MEF pasteurized milk at different electric field strength and
mass flow rates (a: 0.0185 kg/s; b: 0.0425 kg/s and c: 0.0774 kg/s) as compared with those
of raw milk (RM) and thermal pasteurization (TP) during storage. Different letters refer to
significant differences at level 0.05, errors bars represent to standard error.
 Journal Pre-proof

180000 8.33 V/cm 14.58 V/cm 20.8 V/cm TP RM

g

Total count of Bactria (CFU / ml)

160000 f
140000 e
d
120000 c g
b
100000 a f g
e g
80000 f
d e f
60000 c d e
a c d
b c f
40000 a b b
g
a
20000
a e
b c d
0
0 3 6 9 12 15 18
Storage period (day)

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a
8.33 V/cm 14.58 V/cm 20.8 V/cm TP RM

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180000
Total count of Bactria (CFU / ml)

g
160000 f
e
140000
-p d
c g
120000
b g
100000 a
re
f f
e
80000 g
d
60000 c d e e f
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b
a c
40000 d
g
a c
20000 b f
b e
a a b c d
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0
0 3 6 9 12 15 18

b Storage period (day)

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180000 8.33 V/cm 14.58 V/cm 20.8 V/cm TP RM

g
Total count of Bactria (CFU / ml)

160000 f
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140000 e
d
120000 c g
b
a
100000 f
g
80000 e f
c g
60000 b d e f
a b c d
40000 a e g
b d
a c f
20000 e
a b c d
0
0 3 6 9 12 15 18
c Storage period (day)

Figure 8. Total count of bacteria of MEF pasteurized milk at different electric field strength
and mass flow rates (a: 0.0185 kg/s; b: 0.0425 kg/s and b: 0.0774 kg/s) as compared with
those of raw milk (RM) and thermal pasteurization (TP) during storage. Different letters
refer to significant differences at level 0.05, errors bars represent the standard error.
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Table 1. The physical properties of pasteurized milk by thermal pasteurization and

MEF at different flow rates and electric field strengths.
Pasteurizatio EFS TM Density Viscosity FP
MFR
n (V/cm
)kg/s( (°C) (kg/m3) (Pa.s) (°C)
method )
0.018 21.40±0.14 1030.00±5.05 0.00184±0.0003 0.551±0.003
b a
5 0a -a
0.042 21.11±0.09 1031.01±6.96 0.00185±0.0001 0.552±0.001
8.33 b a
5 0a -a
0.077 20.05±0.17 1031.11±8.93 0.00185±0.0006 0.552±0.004
b a
4 1a -a
0.018 1028.03±5.99 0.00182±0.0001 0.541±0.003
22.5±0.26 b b
5 0a -a

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0.042 21.80±0.13 1028.00±7.84 0.00183±0.0003 0.547±0.004
MEF* 14.58 b bc
5 2a -a

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0.077 21.41±0.99 1029.00±9.06 0.00183±0.0001 0.549±0.006
b c
4 1a -a
0.018 22.84±0.78 1025.02±5.98 0.00180±0.0002 0.539±0.003
b
-p d
5 0a -a
0.042 22.10±0.09 1026.11±3.98 0.00181±0.0004 0.547±0.002
20.80
re
5 3b d
0a -a
0.077 21.87±1.07 1027.04±1.99 0.00181±0.0003 0.549±0.001
b b
4 3a -a
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Thermal -
72.01±1.26 1028.00±5.09 0.00188±0.0002
pasteurizatio - - a b a 0.523±0.005
0 a
n
na

-
1031.12±3.87 0.00186±0.0001
RM - - 7.00±0.03 c a 0.552±0.003
1a a
ur

MEF: moderate electric field, RM: raw milk, EFS: electric field strengths, MFR: mass
flow rate, FP: freezing point, TM: the temperature of milk after the process. The
same letters in the column refer to no significant differences at the level of 0.05.
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Table 2. Physicochemical properties of RM, thermal pasteurization, and MEF treated

samples.
MF Moistur
EFS Protei Lacto TA
R e Fat Ash
Sample (V/c n se pH
(kg/ content (%) (%) (%)
m) (%) (%)
s) (%)
0.01 87.02±1 3.61±0 3.7±0. 4.98±0 0.68±0 0.14±0. 6.71±

MEF* 8.33 85 .40a .12 a 20 a .09 a .01 a 03a 0.3 a

0.04 87.02±2 3.62±0 3.71±0 4.96±0 0.69±0 0.14±0. 6.71±

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25 .01 a .14 a .24 a .12 a .03 a 02 a 0.2 a

0.07 87.02±1 3.62±0 3.72±0 4.95±0 0.69±0 0.14±0. 6.71±

74 .05 a .21 a .15 a .16 a .01 a 02 a 0.2 a

0.01 87.01±0 3.58±0 3.70±0 5.03±0 0.68±0 0.14±0. 6.71±

85 .98a .09 a .21 a .12 a .04 a 02 a 0.2 a

14.5 0.04 87.01±1 3.59±0 3.71±0 5.01±0 0.68±0 0.14±0. 6.71±

8 25 .31 a .07 a .17 a .21 a .01 a 03 a 0.3 a

0.07 87.02±2 3.61±0 3.71±0 4.97±0 0.69±0 0.14±0. 6.71

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74 .05 a .10 a .24 a .15 a .03 a 03 a ±0.2a

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0.01 87.01±1 3.58±0 3.70±0 -p 5.04± 0.67±0 0.14±0. 6.71±

85 .56 a .11 a .16 a 0.32a .02 a 03 a 0.2a

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20.8 0.04 87.01±1 3.59±0 3.71±0 5.01±0 0.68±0 0.14±0. 6.71±

0 25 .54 a .15 a .32 a .24 a .05 a 02 a 0.2 a

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0.07 87.01±2 3.60±0 3.71±0 5.00±0 0.68±0 0.14±0. 6.71±

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74 .03 a .16 a .09 a .25 a .02 a 02 a 0.2 a

Thermal
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86.88± 3.57±0 3.70±0 5.18±0 0.67±0 53135± 6.71±

pasteuriz - -
1.04a .25 a .25 a .33 a .03 a 0.03 a 0.2 a
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ation

- 87.02±1 3.62±0 3.72±0 4.94±0 0.70±0 5314±0. 6.71±

RM -
.42 a .18 a .14 a .16 a .01 a 02 a 0.2 a

* MEF: moderate electric field, RM: raw milk, EFS: electric field strengths; MFR:
mass flow rate; TA: titratable acidity. The same letters in the column refer to no
significant differences at the level of 0.05.

Table 3. Microbial counts in milk samples before and after pasteurization by thermal
pasteurization and MEF at different electric field strengths and mass flow rates.
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Yeast TCB
Colifor Staphyloco
Psychrop and reducti
EFS MFR ms ccus
hiles molds TCB on (%)
Alkaline
Sam
(V/c (kg/ (CFU/ aureus
(CFU/mL) (CFU/
(CFU/m phospha
ple mL) (CFU/mL) L) ×104 tase
m) s) ×104 mL)
×104 ×104
×104

0.01 ND ND ND ND 2.91±0. 67.48±1

negative
85 10a .02h

0.04 ND ND ND ND 3.12±0. 65.13±1 negative

8.33 33b .12i
25

0.07 ND ND ND ND 3.23±0. 63.91±2 negative

74 87c .03j

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0.01 ND ND ND ND 1.81±0. 79.77±2 negative
87d .22e

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85

14.5 0.04 ND ND ND ND 2.21±0. 75.30±1 negative

MEF* 67e .42f
8 25
-p
0.07 ND ND ND ND 2.45±0. 72.62±2 negative
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74 90f .09g

0.01 ND ND ND ND 0.73±0. 91.84±2 negative

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85 87i .11a

20.8 0.04 ND ND ND ND 1.12±0. 87.48±1 negative

98g .01b
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0 25

0.07 ND ND ND ND 1.43±0. 84.02±2 negative

74 94k .08c
ur

Thermal ND ND 1.62±0. 81.90±2

N/A N/A negative
68l .02d
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pasteurization
2.31±0 534±0. 8.55±0.
RM 2.16±0.93 0.64±0.88 - positive
.87 003 59m

* MEF: moderate electric field, RM: raw milk, EFS: Electric field strengths, MFR:
mass flow rate; S. aureus: Staphylococcus aureus, TCB: total count Bactria; ND: not
detectable. The same letters in the column refer to no significant differences at the
level of 0.05.

Highlight

 MEF has 63% lower energy consumption compared to the conventional

thermal pasteurization.
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 MEF inactivated alkaline phosphatase by 100%.

 MEF reduced S. aureus from 2.16×104 CFU/mL to non-detectable level at

temperatures below 22°C

 Decontamination enhanced by 55% at the highest field strength (EFS) (20

V/cm) and the minimum mass flow rate (MFR) (0.018 kg/s)

 The longest shelf-life at 5˚C was 15 days that was achieved when EFS and

MFR were 20 V/cm and 0.018 kg/s, respectively.

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Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8