Pulsed Electric Fields' Impact on Milk's Nutritional Composition: History
View Latest Version
Please note this is an old version of this entry, which may differ significantly from the current revision.
Contributor:
Azizah Mohamad
,
Nor Nadiah Abdul Karim Shah
,
Alifdalino Sulaiman
,
Noranizan Mohd Adzahan
,
Rai Naveed Arshad
,
Rana Muhammad Aadil

Milk processing can cause the demolition of the milk fat globule membrane and induce interactions between whey protein and casein with membranes, leading to changes in pH, protein, and lactose content, as well as the destruction of vitamins and enzymes, hydrolysis of proteins and lipids, disruption of calcium and phosphorus equilibrium, and reduction of the cream layer. Pulsed electric fields (PEF) are gaining recognition in food processing due to their energy efficiency, minimal energy loss, flexibility, instantaneity, non-thermal nature, and environmental friendliness. It has also been found to reduce spoilage by microorganisms and the inactivation of undesirable enzymes, as well as its better retention of organoleptic and nutritional characteristics.

  • pulsed electric fields
  • milk
  • macronutrients
  • micronutrients

1. Introduction

Milk is an emulsion of lipid globules and a colloidal mixture of protein and mineral aggregates found in carbohydrate suspension (mostly lactose) [1]. It contains essential nutrients for humans, such as proteins, fats, carbohydrates, vitamins, and minerals [2]. Bovine milk consists of water (~87%), macronutrients that are made up of protein (~3.2%), fat (~3.5%), lactose (~4.8%), and micronutrients, which comprise salts, vitamins, and minerals [3]. Macronutrients provide energy to humans and are needed in large amounts [4], while micronutrients are essential elements that are needed in different amounts by humans to perform various physiological functions to maintain health [5].
Raw milk is a highly nutritious and safe medium for bacterial growth and must be processed appropriately to reduce risks related to public health. The current industrial practice to maintain the safety and shelf-life stability of milk is pasteurization or sterilization via thermal processing. The Food and Drug Administration has published the Grade “A” Pasteurized Milk Ordinance (PMO) to control the standards and regulations on the production, processing, and marketing of milk and dairy products, and this ordinance comprises the comprehensive directives of sanitary facilities, equipment, and practices to ensure the safety and quality of milk [6]. These measures have been effectively used to combat spoilage and pathogenic microbes in milk as well as other food products.
Milk processing can cause the demolition of the milk fat globule membrane and induce interactions between whey protein and casein with membranes, leading to changes in pH, protein, and lactose content, as well as the destruction of vitamins and enzymes, hydrolysis of proteins and lipids, disruption of calcium and phosphorus equilibrium, and reduction of the cream layer [7][8]. Najib et al. [9] proposed that heat-induced acidification in milk leads to the formation of organic acids, insolubility of tertiary calcium phosphate, and casein dephosphorylation. Borad et al. [10] also reported that thermal pasteurization can lead to dehydration, hydrolyzation, or aggregated milk casein. Sharma et al. [11] found that whey proteins are generally thermolabile due to their globular structure, and thermal processing affects milk functionality. This has led to the exploration of non-thermal processing methods such as pulsed electric fields (PEF), high-pressure processing (HPP), and ultrasound (US) as an alternative to complement or substitute the conventional thermal treatment [12][13].
Pulsed electric fields (PEF) are gaining recognition in food processing due to their energy efficiency, minimal energy loss, flexibility, instantaneity, non-thermal nature, and environmental friendliness [14][15]. It has also been found to reduce spoilage by microorganisms and the inactivation of undesirable enzymes, as well as its better retention of organoleptic and nutritional characteristics [16][17][18]. PEF has great potential in the milk processing industry as it causes little change to the flavour, colour, and nutritional value, and is effective in extending the shelf life of milk [8]. Studies have shown that PEF affects different types of milk including bovine milk [11][19][20][21][22][23][24][25][26][27][28][29], goat milk [12][30], and fruit juice–milk beverages [31][32][33][34].

2. Pulsed Electric Fields System

The PEF system consists of a few components, including a repetitive high-voltage pulse generator, treatment chamber, pump or liquid handling system, and control devices [14][35]. Depending on the design of the treatment chamber and the type of treated sample, either a continuous or batch chamber can be operated. A continuous chamber is preferred for industrial applications due to the continuous flow of the samples, while a batch or static chamber can only process the given amount of liquid, solid, or semi-solid samples [14]. Nevertheless, both chambers can achieve similar output and comparable levels of microbial and enzymatic inactivation [14][36][37].
In the PEF process, the sample is positioned in the middle of two or more electrodes before being exposed to high-voltage electric field pulses [38]. PEF can operate at a wide range of ambient temperatures, including ambient (20–25 °C), sub-ambient (<20 °C), and above-ambient (<25 °C) temperatures [39][40], with a short processing time (µs − ms) [18]. PEF can also be applied by varying processing parameters such as electric field strength, pulse width, and pulse frequency, depending on the sample compositions and processing objectives to be acquired [14][18][36][41].
Generally, two mechanisms have been suggested for cell inactivation by PEF, namely electrical breakdown and electroporation [18]. Electrical breakdown occurs when the PEF achieves a greater electric field strength than the critical field strength of the microorganism, resulting in direct discharge and the breaking down of the cell membranes [42]. The biological cells are an electrolyte enclosed by an electrically injured membrane, which is exposed to an extrinsic electric field and subsequently induces transmembrane voltage [38]. The possible outcomes of cell electroporation are shown in Figure 1. Electroporation is the process of temporarily weakening the lipid bilayer and proteins of the cell membranes, resulting in the transformation of cell membrane permeability and the subsequent draining of the membrane [43][44]. It can be reversible or irreversible, depending on the PEF processing parameter, medium constituents, type, and size of the cells [14]. PEF has been used in a variety of food processing applications such as meat processing [45] and the retention of bioactive compounds from fruits and vegetables [46]. Although irreversible, PEF is used for microbial and enzyme inactivation in liquid foods such as milk and fruit juices [30][47].
Foods 12 02114 g001
Figure 1. Cell electroporation outcomes are dependent on the pulsing dynamic (amplitude, shape, and duration of pulses) and other cell manipulation approaches (modified and adapted from [42]).

3. Effect of Pulsed Electric Fields on Milk Nutrients

Table 1, Table 2, Table 3 and Table 4 summarize the effects of PEF on fats and fatty acids, proteins and amino acids, vitamins, and minerals in milk, respectively.
Table 1. Effects of PEF on the fats and fatty acids in milk.
Nutrient Constituents Products PEF Parameter Effects References
Fats and fatty acids Raw bovine milk and milk with different MFG sizes 16 kV/cm; 30 µs; 100 mL/min; 25 °C Increase in FAs and some unknown long-chain FAs in raw milk and milk with large MFGs; a slight decrease in the fat content [19]
Fatty acid profiles Raw goat milk 20, 30, and 40 kV/cm; 5 and 10 μs; 2.5 L/h; 30 °C Reduction in the total SFAs and total PUFAs; increase in total MUFAs [30]
Fatty acids Fruit juices-milk beverages 35 kV/cm; 1800 μs; 60 mL/min; 60 mL/s; <40 °C 20% increase in linoleic acid; decrease in palmitic, linoleic, and linolenic acids (12–20%) at day 56 [31]
Fatty acid profiles Raw whole bovine milk (4% fat) 30 kV/cm; 22 μs; 2.4 L/min; 53 and 63 °C No significant changes (p > 0.05) in fat content; minor increase in butanoic (C4:0), hexanoic (C6:0), and octanoic (C8:0) [22]
Fats Bovine whole milk 20 and 26 kV/cm; 34 μs; 4.2 mL/s; 55 °C Increase fat melting from a-crystal into b-crystal conversion [20]
MFG Bovine whole milk (44% fat) 20 and 26 kV/cm; 34 μs; 4.2 mL/s; 17–22 °C Reduction in MFGs’ size; increase in ζ-potential and specific surface area; plasma protein absorption on MFGM surface [21]
Milk fat Reconstituted skimmed milk 15–20 kV/cm; 20–60 pulses; <35 °C Increase the surface area of MFGs [27]
Volatile compounds (including fatty acids) Raw bovine milk 15–30 kV/cm; 800 µs; <40 °C No significant changes (p > 0.05) in methyl ketones [saturated fatty acids); no changes in concentration of short-chain fatty acid [29]
Milk fat globule membrane (MFGM), fatty acid (FA), saturated fatty acid (SFA), monounsaturated fatty acid (MUFA), polyunsaturated fatty acid (PUFA).
Table 2. Effects of PEF on the milk proteins and amino acids.
Nutrient Constituents Products PEF Parameter Effects References
Proteins Raw bovine milk and milk with different MFG sizes 16 kV/cm; 30 µs; 100 mL/min; 25 °C 15% and 29% decrease in β-Lg and α-La respectively (in milk with small MFGs); more small protein aggregates (<40 kDa) appeared [19]
Native proteins 10% solution of liquid whey protein 50 kV/cm; 3–8 µs; 5 L/h; 20–40 °C Stable IgM and IgG [48]
Whey proteins Bovine whole milk 20 and 26 kV/cm; 34 μs; 4.2 mL/s; 55 °C Increase in the hydrophobic surface of the protein; reducion in milk protein denaturation [20]
Caseins and whey proteins Reconstituted skim milk (10 wt%) 49 kV/cm; 19.36 µs; 240 mL/min; 20–72 °C Increase in the amount of protein in the serum; increase the number of caseins in serum; no changes in whey proteins [23]
Protein compositions Fresh raw cream and pasteurized cream (40% fat) 37 kV/cm; 1705 μs;
25 mL/min; 50 and 65 °C		 No changes in phospholipids; induced interactions of β-Lg
and MFGM proteins (at 65 °C).		 [49]
Milk proteins Raw whole milk 40 kV/cm; <15 s; 30 L/h; 72 °C 70% decrease in the native form of milk proteins including IgG, IgA, and lactoferrin [24]
Protein BSA 20–35 kV/cm; 400 μs; 200 Hz; <30 °C No self-aggregation of BSA; no significant difference (p > 0.05) in molecular weight distribution profiles of BSA [50]
Protein content Raw skim and raw whole milk 30.76–53.84 kV/cm; 12–30 pulses, 1 L/min; 20–40 °C A 0.17% decrease in protein content [26]
Whey protein Whey protein isolate 12–20 kV/cm; 10–20 pulses; <35 °C Increase polarity and unfolding whey proteins; partial denaturation of whey fractions [51]
Protein aggregation Whey protein isolate 30–35 kV/cm; 19.2–211 µs; 60 mL/min; 30–75 °C No changes in protein aggregation and surface hydrophobicity; partial denaturation of native lactoferrin, apo-lactoferrin, and halo-lactoferrin [52]
Milk casein micelles’ sizes Raw milk, reconstituted skim milk, concentrated skim milk, and milk protein concentrates 45 kV/cm; 20 µs; 240 mL/min; 30 °C Decrease in casein micelles’ sizes [28]
Proteolysis profiles (peptide and free amino acid concentration) Raw milk cheese 30 kV/cm; 80–120 pulses; 6 mL/min; 50 °C). Intermediate proteolysis profiles between raw milk and thermally pasteurized milk
Severe enzyme modification reduces the formation of peptides and amino acids at longer pulses		 [53]
Milk fat globule membrane (MFGM), bovine serum albumin (BSA), β-lactoglobulin (β-Lg), α-lactalbumin (α-La), immunoglobulin M (IgM), immunoglobulin G (IgG).
Table 3. Effects of PEF on the milk vitamins.
Nutrient Constituents Products PEF Parameter Effects References
Vitamin A (as β-carotene) and B-group vitamins (thiamine, riboflavin, and niacin) Raw goat milk 20, 30, and 40 kV/cm; 5–13 µs; 2.5 L/min; 30 °C >94% retention of vitamin A (as β-carotene); 95–99% retention of B-group vitamins [12]
Vitamins A and C 10% solution of liquid whey protein 50 kV/cm; 3–8 µs; 5 L/h; 20–40 °C No changes in vitamins A and C [48]
Total carotenoids (provitamin A), vitamin C, in vitro bioaccessibility of vitamin C Fruit juice and milk beverages 35 kV/cm; 1800 µs; 60 mL/min; <35 °C 15% increase in the bioaccessibility of total carotenoids (provitamin A); 8–15% reduction of vitamin C; no changes in bio-accessibility of vitamin C [32]
Total carotenoids (provitamin A), vitamin C (ascorbic acid) Orange juice-milk beverage 25 kV/cm; 280 µs; 60 mL/min; 57 °C 2% destruction of ascorbic acid;5% destruction of total carotenoids [33]
B-group vitamins (niacin, thiamin, and riboflavin), vitamin C FJ-WM and FJ-SM beverages 35 kV/cm; 1800 µs; 760 mL/min; <40 °C 95–99.8% retention of group B vitamins (niacin, thiamin, and riboflavin); 97% retention of vitamin C in FJ-WM; 99.5% retention of vitamin C in FJ-SM [34]
Fruit juice–whole milk (FJ–WM), fruit juice–skim milk (FJ–SM).
Table 4. Effects of PEF on the milk minerals.
Nutrient Constituents Products PEF Parameter Effects References
Fe, Cr, Ni, and Mn Raw goat milk 20, 30, and 40 kV/cm; 5 and 10 μs; 2.5 L/h; 30 °C Significant increase (p < 0.05) in Fe;
A non-significant increase in (p > 0.05) in Cr, Ni, and Mn		 [30]
Fe, Cr, and Ni 2% partially skimmed milk 30–40 kV/cm; 500 mL/min; 20–25 °C A significant increase (p < 0.05) in Fe [25]

3.1. Fats and Fatty Acids

Fats or lipids are the primary elements of milk that influence the cost, nutrition, functional, and sensory attributes of dairy products [54]. A milk fat globule membrane (MFGM) comprises polar lipids, cholesterol, proteins, glycoproteins, and enzymes [22]. The composition and structure of milk fat globules (MFG) and MFGM and their composition contribute to lipases’ access to lipids, which can result in structural changes in the MFGM [19]. Therefore, it is essential to review the effect of PEF processing on the modification of the fatty acid compositions in milk.
The temperature has a significant effect on determining the rate of lipid oxidation since fats and FAs are susceptible to oxidation [55]. Lipid oxidation products are thought to be a risk factor for human health because of their mutagenic, carcinogenic, and cytotoxic characteristics [56]. Temperature influences chemical reactivity, colloidal structure organization, and stability, as well as the rate of synthesis and decomposition of hydroperoxides [57]. Despite the fact that PEF is a non-thermal technique, ohmic heating can cause temperatures to increase [12]. Ohmic or joule heating in the PEF process emerges when an electric current flows through a sample and ions and other charged molecules, such as lipids and proteins, take on the opposite charge of the electrode [56].
Earlier, McAuley et al. [22] demonstrated that PEF treatment (30 kV/cm; 22 μs; 2.4 L/min; 53 and 63 °C) had no significant effect on fat, protein, lactose, total solids, and the total solids non-fat of raw whole milk (4% fat). However, the authors observed a minor increase in the milk lipid-derived short-chain FFAs, namely butanoic (C4:0), hexanoic (C6:0), and octanoic (C8:0) acids, during two weeks of refrigerated storage at 4 and 8 °C, due to the greater rate of milk triglyceride hydrolysis by milk-derived lipases and/or esterases (lipoprotein lipase, LPL) and milk microbiota [22]. LPL is the main native milk enzyme. It only becomes active on the fat in natural MFGs when physical treatments have broken them down or if certain blood serum lipoproteins are present [58]. Nevertheless, they propose that the LPL activity is typically less active in milk fat because it is being sheltered by the MFGM and glycoproteins in the skim fraction that inhibit the lipolysis process. Similarly, the changes in FA profiles were also observed by other researchers [30]. They found a significant reduction (p < 0.05) in total SFAs and total PUFAs, while the number of total MUFAs increased in PEF-treated goat milk (20, 30, and 40 kV/cm; 5 and 10 μs; 2.5 L/h; 30 °C). The authors suggest that the changes in FA may have been caused by the breakdown of fat and triacylglycerol from MFGM and the catalytic oxidation of heterogeneous metals (Fe and Ni) from the PEF electrode material, which resulted in significant decreases in various FAs. Moreover, some other studies reported that nickel showed the best isomerization properties for FA [59]. According to Ghnimi et al. [57], heterogeneous catalysis by metal oxides can initiate FA oxidation by generating micelles comprising primary hydroperoxide, water, and other amphiphilic compounds. They also suggest that the metal oxide can provide a surface site catalyst for hydrocarbon deprotonation on the nanoscale since the C–H bonds of hydrocarbons are very weakly acidic and unfit for hydrocarbon deprotonation. Despite that, further investigation is needed to understand the PEF electroporation and heterogeneous metal catalysis effects on fatty acid molecules.
According to Yang et al. [19], PEF treatment at 16 kV/cm; 30 µs; 100 mL/min; 25 °C slightly reduced the fat contents and FFA of three types of milk samples: raw milk (RM), milk with small fat globules (MS), and milk with large fat globules. Nevertheless, the same authors found that the MS with the smallest MFGs and the most modified MFGM exhibited a minor increase in FFA following the same PEF treatment. The authors proposed that the changes in FFA are likely due to the electroporation of MFGMs that exposed triglycerides to lipases and subsequently led to the release of FFA. These findings concur well with those of Garcia-Amezquita et al. [60], who found that PEF processing (36 and 42 kV/cm; 2.6 µs; 23 L/h; 25 °C) led to the clustering of small milk-fat globules, which appeared to increase the population of larger globules. However, the actual mean diameter of MFG was unaffected by PEF. It has been reported that PEF processing may result in oxidation (loss) and/or reduction (gain) in both the electrode and the electrolyte (sample) due to electrochemical reactions that took place when the interface voltage exceeded the threshold voltage [30][61]. Therefore, the decrease in fat content can be attributed to the partial loss of some fat clumps that are attached to the PEF tube or electrode.

3.2. Proteins and Amino Acids

Milk proteins consist of an unstable micellar phase, known as casein, and a soluble phase of whey proteins [54]. The casein phase contains approximately 80% of the proteins and is classified as αs1, αs2, β, and κ-caseins [62]. Whey-phase proteins with the highest concentrations are α-lactalbumin (α-La) and β-lactoglobulin (β-Lg) [63]. Milk contains approximately 3.5 g/100 mL of high-quality protein, comprising nine essential amino acids required by humans [64]. Furthermore, protein components are present in the MFGM as lipoproteins, with casein having the highest content (~80%) of proteinaceous substances in bovine milk, followed by whey protein (~20%); this ratio varies by species [64].
Generally, PEF treatment affects the structure of proteins, especially the secondary and tertiary structures [65] leading to denaturation and consequent changes in their functional properties such as solubility, gelation, and aggregation. PEF has been described to affect the structure of proteins and amino acids, which are primarily associated with weaker bonds such as disulfide bonds, hydrogen bonds, and hydrophobic bonds [66]. In addition, disulfide bonds are the main covalent bonds resulting from the PEF-induced protein aggregates [50]. As a result, PEF may promote protein augmentation via hydrophobic and thiol or disulfide reaction aggregation, altering protein thermal stability and enzymatic digestibility [67]. Additionally, it has been demonstrated that some chemical bonds, such as peptides and non-covalent bonds, as well as electrostatic, hydrophobic, and Van der Waals interactions, counterbalance the protein structure [68]. The author also suggests that there is a considerable chance that the application of an electric field may affect the conformation of proteins, due to the presence of electrostatic forces in the peptide chain. These have a good correlation with Yu et al. [53], who found that the alteration of the enzyme by PEF may have reduced the proteolytic enzyme characteristics of milk, resulting in less peptide and amino acid production in PEF-treated milk (30 kV/cm; 80–120 pulses; 6 mL/min; 50 °C). The author also discovered that an increase in pulses may cause an enzyme modification to occur more severely, and that a decrease in enzyme activity may cause a decrease in the production of peptides and free amino acids.
Furthermore, Yang et al. [19] observed that the β-Lg band in milk with small fat globules (MS) was reduced following PEF treatments by 8% (9 kV/cm; 30 µs; 100 mL/min; 25 °C) and 15% (16 kV/cm; 30 µs; 100 mL/min; 25 °C), whereas raw milk (RM) and milk with large fat globules (ML) demonstrated no significant changes in the β-Lg band. Likewise, the band for α-La in MS was similarly reduced following PEF treatments by 11% and 26% at 9 and 16 kV/cm, respectively (30 µs; 100 mL/min; 25 °C); but α-La in RM showed no changes and ML showed just a minor decrease (2–5%). The authors suggest that following PEF treatments, β-Lg and α-La were primarily adsorbed to the small MFGs in MS. They also found that the primary MFGM proteins, xanthine oxidase/dehydrogenase (XO/XDH), were present in the RM sample. The ML sample has a comparable profile for the MFGM proteins; however, the MS sample exhibits a very different profile since it almost completely lacked the XO/XDH band and had additional lower molecular weight protein bands as well as numerous fuzzy protein aggregation bands [19]. The authors also suggest that the MFGM composition of small MFGs differs significantly from that of large MFGs in ML, proving that the membrane of the small MFGs underwent more profound modifications throughout the separation process.
Other studies have suggested that PEF treatment can alter protein polarity and the microenvironment of amino acid residues due to the liberation of native molecular structures, including the whey protein structure [51]. The changes in the milk protein were also observed by Mathys et al. [24], who found that PEF treatment (40 kV/cm; <15 s; 30 L/h; 20–45 °C) reduced 70% of the native form of milk proteins, including immunoglobulin G (IgG), immunoglobulin A (IgA), and lactoferrin. Moreover, the authors affirm that the variations in milk proteins correspond to the specific energy of PEF treatment. This suggests that high intensity of PEF treatment (>30 kV/cm) along with a mild temperature (40–60 °C) can induce denaturation of milk proteins, possibly due to the unfolding of protein molecules or their positioning towards an applied electric field. These correlate favourably with Sui et al. [52], who perceived significant changes in the concentration of natively folded lactoferrin, aggregated protein, and surface hydrophobicity of whey protein isolate after PEF treatment (30–35 kV/cm, 19.2 and 211 µs; 760 mL/min; 60–70 °C).
Accordingly, Sharma et al. [69] also reported that the high intensity of PEF could increase the PEF processing temperatures due to higher specific energy, which may lead to the unfolding of protein molecules, the alignment of proteins towards electric fields, and transitions in the structure of the native proteins. They proposed that PEF-induced protein conformational changes are due to denaturation and the formation of intermolecular structures such as β-Lg or κ-casein following cross-linking between casein and serum proteins. In another study, the same authors [21] observed a decrease in MFG size, an increase in ζ-potential, specific surface area, and plasma protein absorption on the MFGM surface of PEF-treated bovine whole milk (20 and 26 kV/cm; 34 μs; 4.2 mL/s; 17–19.4 °C). It has been asserted that the superficial coverage of MFGM through plasma proteins was associated with the increase in ζ-potential, where the ζ-potential is an indication of the degree of damage to the surface of MFGM [70]. Moreover, the decrease in the size of MFG may be attributable to the shear force effect during pumping. These are in agreement with Hemar et al. [28], who reported the shear effect produced during circulating milk through the PEF system that can lead to a partial separation of casein micelles. However, the authors claimed that the effects of higher temperatures (63 °C for 30 min; 73 °C for 15 s) outweighed the modifications brought about by PEF treatment.
In contrast, Liu et al. [23] found that PEF treatment (49 kV/cm; 19.36 s; 240 mL/min; 20 °C) did not affect the physicochemical properties of milk caseins, the size and ζ-potential of milk particles, the status of the whey protein, or the amount of protein in the serum. The variable medium (sample), processing settings, and PEF system used in the study can be blamed for this rather paradoxical outcome. Nevertheless, these findings appear to be well substantiated by Schottroff et al. [48], who obtained stable immunoglobulin M (IgM) and immunoglobulin G (IgG) in the 10% solution of liquid whey protein treated with PEF (50 kV/cm; 3–8 µs; 5 L/h; 20–40 °C). Since the majority of whey proteins are more susceptible to heat denaturation at neutral pH than under acidic circumstances, they contend that PEF is unlikely to have an impact on soluble proteins. Additionally, the results are consistent with Sharma et al. [20], who discovered that the PEF treatment (20 and 26 kV/cm; 34 s; 4.2 mL/s; 55 °C) had no appreciable impacts on the endothermic peak of bovine whole milk at 32.2 °C. According to this, PEF could maintain protein functioning and integrity by lowering milk protein denaturation, which would lead to less absorption onto the surface of MFGM.

3.3. Vitamins

Vitamins are a group of organic complexes that are needed in small quantities; however, a lack of vitamins will lead to such syndromes as scurvy and beriberi. Vitamins fall into two categories: those that are fat-soluble (such as vitamins A, D, E, and K) and others that are water-soluble (such as vitamins B and C). It has been proposed that many variables, including light, temperature, pH, oxygen availability, metal catalysts, and the inclusion of additional antioxidants or reducing agents, can affect the stability of vitamins [68].
According to earlier research, PEF treatment had a negligible impact on milk’s vitamin content [12][33][34][48][71]. Mohamad et al. [12] discovered no significant changes (p > 0.05) in vitamin A (as β-carotene) and B-group vitamins (thiamine, riboflavin, and niacin) in PEF-treated goat milk (20–40 kV/cm; 5–13 s; 2.5 L/min; 30 °C). Since vitamin A and B-group vitamins are known to be sensitive to a higher temperature, the higher retention value (>94%) observed in the study is likely attributable to the lower PEF processing temperature (30 °C) used in the study. The authors conclude that extended PEF treatment time and high electric field intensities lead to decreased vitamin retention values. Riener et al. [71] reported that PEF treatment (15–35 kV/cm; 12.5–75 s; 30 °C) had no significant effects (p > 0.05) on the levels of thiamin, riboflavin, retinol, and α-tocopherol in fresh bovine raw milk. The effect of PEF (18.3–27.1 kV/cm; 400 μs; 20–25 °C) on both water-soluble (thiamin, riboflavin, and ascorbic acid) and fat-soluble (cholecalciferol and tocopherol) vitamins in milk has been evaluated by Bendicho et al. [72]. The authors observed no significant changes (p > 0.05) in both classes of vitamins except for ascorbic acid. Likewise, the reduction of ascorbic acid was found to be dependent on the electric field strength, treatment time, and temperature. However, they argued that PEF provides better retention of ascorbic acid (vitamin C) compared to thermal treatment (63 and 75 °C; 15 s and 30 min).
PEF treatment has also been demonstrated to effectively maintain the bioaccessibility of provitamin A (carotenoids) from fruit juices and milk beverages. For instance, Rodríguez-Roque et al. [32] observed that PEF treatment (35 kV/cm; 1800 s; 60 mL/min; 35 °C) increased the bioaccessibility of total carotenoids in fruit juices (orange, pineapple, kiwi, and mango) and milk beverages by 15%. They found that PEF treatment gives greater stability to the total carotenoids due to the inactivation of oxidative enzymes, the interruption of cell membranes and proteins, and the release of some individual carotenoids. These are consistent with Zulueta et al. [33], who obtained minimal losses of ascorbic acid and total carotenoids with only 2 and 5% destruction rates, respectively, in PEF-treated orange juice-milk beverages (25 kV/cm; 280 µs; 60 mL/min; 57 °C). Salvia-Trujillo et al. [34] also observed higher retention (95–99%) of group B vitamins (niacin, thiamin, and riboflavin) in PEF-treated fruit juice–whole milk (FJ–WM) and fruit juice–skim milk (FJ–SM) beverages (35 kV/cm; 1800 µs; 760 mL/min; <40 °C). Following this, they discovered increased vitamin C retention (>97%) in PEF-treated FJ–WM and FJ–SM beverages, as opposed to 50% retention following 21 days of storage at 4 °C. The results demonstrate that water-soluble vitamins, particularly niacin, thiamin, and riboflavin, were not oxidatively degraded by PEF. They demonstrate that water-soluble vitamins are more responsive to heat treatment than to PEF treatment by contrasting heat (90 °C) and PEF treatment (40 °C). These findings are in good agreement with those of Schottroff et al. [48], who reported that after PEF treatment (50 kV/cm; 3–8 s; 5 L/h; 20–40 °C), there were no significant losses (p > 0.05) of vitamins A and C in liquid whey protein. The findings would seem to indicate that PEF treatment does not subject vitamins to any chemical changes.

3.4. Minerals

Milk contains 20 minerals that are necessary for human nutrition. The two groups of these minerals are called macro- and microminerals. These macro- and microminerals occur in milk in an equilibrium between the soluble and colloidal phases, each serving a different physiological purpose [73]. For instance, Na and K are crucial in preserving the osmotic equilibrium between blood and milk, while Mg, Ca, P, and Zn are attached to casein micelles in milk, making their content directly tied to casein content [74]. It has been reported that Mg varies by species and is frequently connected to milk protein [75]. Although milk only contains 8–9 g/L of minerals, it is nevertheless an important source of nutrition [76].
The impact of PEF on the mineral profiles of milk has not yet been extensively researched. A literature search revealed that there is still some pertinent research on the effects of PEF on milk minerals in other species of animals. For instance, Mohamad et al. [31] found that the Fe levels in goat milk treated with PEF increased significantly (p < 0.05) at 20–40 kV/cm, 5–10 s, 2.5 L/h, and 30 °C. However, after PEF treatment, it was determined that the other minerals (Cr, Ni, and Mn) were not significant (p > 0.05). The authors concluded that the electrochemical reaction that takes place during PEF treatment may have caused the metal components from the PEF electrode to dissolve into the sample. The results obtained are in close agreement with those of Gad and Jayaram [25], who investigated the release of metal particles from stainless-steel electrodes in samples of 2% partially skimmed milk. The Fe concentration rose by 500 µg/L following PEF treatment (30–40 kV/cm; 500 mL/min; 20–25 °C) as opposed to 300 µg/L in untreated samples. The authors agreed that the commercialization of PEF processing may be impacted by electrochemical reactions that happen during milk processing about safety and quality.

This entry is adapted from the peer-reviewed paper 10.3390/foods12112114

References

  1. Altomonte, I.; Salari, F.; Martini, M. Milk fat components and milk quality. In Microbiology in Dairy Processing: Challenges and Opportunities; John Wiley & Sons, Inc.: Hoboken, NJ, USA, 2017; Volume 8, pp. 1–10.
  2. Yang, S.; Liu, G.; Qin, Z.; Munk, D.; Otte, J.; Ahrne, L. Effect of Emerging Processing Methods on the Food Quality: Advantages and Challenges; Springer: Berlin/Heidelberg, Germany, 2019; pp. 27–67.
  3. O’Callaghan, T. The benefits of pasture-based dairy. Ir. Food 2019, 1, 34–35.
  4. Zheroz. Importance of Micronutrients and Macronutrients in Your Daily Diet. Available online: https://zheroz.wordpress.com/2018/09/19/importance-of-micronutrients-and-macronutrients-in-your-daily-diet/ (accessed on 1 September 2022).
  5. Gernand, A.D.; Schulze, K.J.; Stewart, C.P.; West, K.P., Jr.; Christian, P. Micronutrient deficiencies in pregnancy worldwide: Health effects and prevention. Nat. Rev. Endocrinol. 2016, 12, 274–289.
  6. Lu, C.D.; Miller, B.A. Current status, challenges and prospects for dairy goat production in the Americas. Asian-Australas. J. Anim. Sci. 2019, 32, 1244–1255.
  7. Sakkas, L.; Moutafi, A.; Moschopoulou, E.; Moatsou, G. Assessment of heat treatment of various types of milk. Food Chem. 2014, 159, 293–301.
  8. Khanam, T.G.; Goswami, M.; Pathak, V.; Bharti, S.K.; Karunakara, K.N. Pulse electric field: Novel approach to dairy industry. Indian Food Ind. Mag. 2017, 36, 8–12.
  9. Najib, M.; Hallab, M.W.; Hallab, K.; Hallab, Z.; Hamze, M.; Chihib, N. Thermal processing of milk as a main tool in the production of Qishta, Khoa and Kajmak. J. Mater. Environ. Sci. 2020, 11, 294–309.
  10. Borad, S.G.; Kumar, A.; Singh, A.K. Effect of processing on nutritive values of milk protein. Crit. Rev. Food Sci. Nutr. 2017, 57, 3690–3702.
  11. Sharma, P.; Oey, I.; Bremer, P.; Everett, D.W. Microbiological and enzymatic activity of bovine whole milk treated by pulsed electric fields. Int. J. Dairy Technol. 2017, 71, 10–19.
  12. Mohamad, A.; Shah, N.N.A.K.; Sulaiman, A.; Mohd Adzahan, N.; Aadil, R.M. Impact of the pulsed electric field on physicochemical properties, fatty acid profiling, and metal migration of goat milk. J. Food Process. Preserv. 2021, 44, e14940.
  13. Scudino, H.; Silva, E.K.; Gomes, A.; Guimarães, J.T.; Cunha, R.L.; Sant’Ana, A.S.; Meireles, M.A.A.; Cruz, A.G. Ultrasound stabilization of raw milk: Microbial and enzymatic inactivation, physicochemical properties and kinetic stability. Ultrason. Sonochemistry 2020, 67, 105185.
  14. Arshad, R.N.; Abdul-Malek, Z.; Munir, A.; Buntat, Z.; Ahmad, M.H.; Jusoh, Y.M.M.; El-Din Bekhit, A.; Roobab, U.; Manzoor, M.F.; Aadil, R.M. Electrical systems for pulsed electric field applications in the food industry: An engineering perspective. Trends Food Sci. Technol. 2020, 104, 1–13.
  15. Pal, M. Pulsed Electric Field Processing: An Emerging Technology for Food Preservation. J. Exp. Food Chem. 2017, 3, 1000126.
  16. Nowosad, K.; Sujka, M.; Pankiewicz, U.; Kowalski, R. The application of PEF technology in food processing and human nutrition. J. Food Sci. Technol. 2021, 58, 397–411.
  17. Ahmad, T.; Butt, M.Z.; Aadil, R.M.; Inam-Ur-Raheem, M.; Bekhit, A.E.D.; Guimarães, J.T.; Balthazar, C.F.; Rocha, R.S.; Esmerino, E.A.; Freitas, M.Q. Impact of nonthermal processing on different milk enzymes. Int. J. Dairy Technol. 2019, 72, 481–495.
  18. Roobab, U.; Aadil, R.M.; Madni, G.M.; Bekhit, A.E.D. The impact of nonthermal technologies on the microbiological quality of juices: A review. Compr. Rev. Food Sci. Food Saf. 2018, 17, 437–457.
  19. Yang, S.; Suwal, S.; Andersen, U.; Otte, J.; Ahrné, L. Effects of pulsed electric field on fat globule structure, lipase activity, and fatty acid composition in raw milk and milk with different fat globule sizes. Innov. Food Sci. Emerg. Technol. 2021, 67, 102548.
  20. Sharma, P.; Oey, I.; Everett, D.W. Thermal properties of milk fat, xanthine oxidase, caseins and whey proteins in pulsed electric field-treated bovine whole milk. Food Chem. 2016, 207, 34–42.
  21. Sharma, P.; Oey, I.; Everett, D.W. Interfacial properties and transmission electron microscopy revealing damage to the milk fat globule system after pulsed electric field treatment. Food Hydrocoll. 2015, 47, 99–107.
  22. McAuley, C.M.; Singh, T.K.; Haro-Maza, J.F.; Williams, R.; Buckow, R. Microbiological and physicochemical stability of raw, pasteurised or pulsed electric field-treated milk. Innov. Food Sci. Emerg. Technol. 2016, 38, 365–373.
  23. Liu, Z.; Hemar, Y.; Tan, S.; Sanguansri, P.; Niere, J.; Buckow, R.; Augustin, M.A. Pulsed electric field treatment of reconstituted skim milks at alkaline pH or with added EDTA. J. Food Eng. 2015, 144, 112–118.
  24. Mathys, A.; Toepfl, S.; Siemer, C.; Favre, L.; Benyacoub, J.; Hansen, C.E. Pulsed Electric Field Treatment Process and Dairy Product Comprising Bioactive Molecules Obtainable by the. Process. Patent No WO2013007620A1, 17 January 2013.
  25. Gad, A.; Jayaram, S.H. Effect of electric pulse parameters on releasing metallic particles from stainless steel electrodes during PEF processing of milk. IEEE Trans. Ind. Appl. 2013, 50, 1402–1409.
  26. Bermúdez-Aguirre, D.; Fernández, S.; Esquivel, H.; Dunne, P.C.; Barbosa-Cánovas, G.V. Milk processed by pulsed electric fields: Evaluation of microbial quality, physicochemical characteristics, and selected nutrients at different storage conditions. J. Food Sci. 2011, 76, S289–S299.
  27. Xiang, B.; Simpson, M.; Ngadi, M.; Simpson, B. Flow behaviour and viscosity of reconstituted skimmed milk treated with pulsed electric field. Biosyst. Eng. 2011, 109, 228–234.
  28. Hemar, Y.; Augustin, M.A.; Cheng, L.J.; Sanguansri, P.; Swiergon, P.; Wan, J. The effect of pulsed electric field processing on particle size and viscosity of milk and milk concentrates. Milchwiss.-Milk Sci. Int. 2011, 66, 126.
  29. Zhang, S.; Yang, R.; Zhao, W.; Hua, X.; Zhang, W.; Zhang, Z. Influence of pulsed electric field treatments on the volatile compounds of milk in comparison with pasteurized processing. J. Food Sci. 2011, 76, C127–C132.
  30. Mohamad, A.; Abdul Karim Shah, N.N.; Sulaiman, A.; Mohd Adzahan, N.; Aadil, R.M. Pulsed electric field of goat milk: Impact on Escherichia coli ATCC 8739 and vitamin constituents. J. Food Process Eng. 2021, 44, e13779.
  31. Salvia-Trujillo, L.; Morales-de la Pena, M.; Rojas-Grau, A.; Welti-Chanes, J.; Martin-Belloso, O. Mineral and fatty acid profile of high intensity pulsed electric fields or thermally treated fruit juice-milk beverages stored under refrigeration. Food Control 2017, 80, 236–243.
  32. Rodriguez-Roque, M.J.; de Ancos, B.; Sanchez-Vega, R.; Sanchez-Moreno, C.; Cano, M.P.; Elez-Martineza, P.; Martin-Belloso, O. Food matrix and processing influence on carotenoid bioaccessibility and lipophilic antioxidant activity of fruit juice-based beverages. Food Funct. 2016, 1, 380–389.
  33. Zulueta, A.; Barba, F.J.; Esteve, M.J.; Frigola, A. Changes in quality and nutritional parameters during refrigerated storage of orange juice-milk beverage treated by equivalent thermal and non-thermal processes for mild pasteurization. Food Bioprocess Technol. 2013, 6, 2018–2030.
  34. Salvia-Trujillo, L.; Morales-de la Pena, M.; Rojas-Grau, A.; Martin-Belloso, O. Changes in water-soluble vitamins and antioxidant capacity of fruit juice-milk beverages as affected by high-intensity pulsed electric fields (HIPEF) or heat during chilled storage. J. Agric. Food Chem. 2011, 59, 10034–10043.
  35. Gómez, B.; Munekata, P.E.; Gavahian, M.; Barba, F.J.; Martí-Quijal, F.J.; Bolumar, T.; Campagnol, P.C.B.; Tomasevic, I.; Lorenzo, J.M. Application of pulsed electric fields in meat and fish processing industries: An overview. Food Res. Int. 2019, 123, 95–105.
  36. Chemat, F.; Rombaut, N.; Meullemiestre, A.; Turk, M.; Perino, S.; Fabiano-Tixier, A.-S.; Abert-Vian, M. Review of green food processing techniques. Preservation, transformation, and extraction. Innov. Food Sci. Emerg. Technol. 2017, 41, 357–377.
  37. Manzoor, M.F.; Zeng, X.A.; Ahmad, N.; Ahmed, Z.; Rehman, A.; Aadil, R.M.; Roobab, U.; Siddique, R. Effect of pulsed electric field and thermal treatments on the bioactive compounds, enzymes, microbial and physical stability of almond milk during storage. J. Food Process. Preser. 2020, 44, e14541.
  38. Toepfl, S.; Heinz, V.; Knorr, D. High intensity pulsed electric fields applied for food preservation. Chemical engineering and processing. Process Intensif. 2007, 46, 537–546.
  39. Elez-Martínez, P.; Odriozola-Serrano, I.; Oms-Oliu, G.; Soliva-Fortuny, R.; Martín-Belloso, O. Effects of pulsed electric fields processing strategies on health-related compounds of plant-based foods. Food Eng. Rev. 2017, 9, 213–225.
  40. Syed, Q.A.; Ishaq, A.; Rahman, U.U.; Aslam, S.; Shukat, R. Pulsed electric field technology in food preservation: A review. J. Nutr. Health Food Eng. 2017, 6, 168–172.
  41. Arshad, R.N.; Abdul-Malek, Z.; Roobab, U.; Qureshi, M.I.; Ahmad, M.H.; Malik, N.; Bekhit, A.E.D.; Liu, Z.W.; Aadil, R.M. Pulsed electric field: A potential alternative towards a sustainable food processing. Trends Food Sci. Technol. 2021, 111, 43–54.
  42. Ozturk, B.; Anli, E. Pulsed electric fields (PEF) applications on wine production: A review. In BIO Web of Conferences; EDP Sciences: Les Ulis, France, 2017; p. 02008.
  43. Liu, Z.; Zhao, L.; Zhang, Q.; Huo, N.; Shi, X.; Li, L.; Jia, L.; Lu, Y.; Peng, Y.; Song, Y. Proteomics-based mechanistic investigation of Escherichia coli inactivation by pulsed electric field. Front. Microbiol. 2019, 10, 2644.
  44. Montanari, C.; Tylewicz, U.; Tabanelli, G.; Berardinelli, A.; Rocculi, P.; Ragni, L.; Gardini, F. Heat-assisted pulsed electric field treatment for the inactivation of Saccharomyces cerevisiae: Effects of the presence of citral. Front. Microbiol. 2019, 10, 1737.
  45. Alahakoon, A.U.; Oey, I.; Bremer, P.; Silcock, P. Process optimisation of pulsed electric fields pre-treatment to reduce the sous vide processing time of beef briskets. Int. J. Food Sci. Technol. 2019, 54, 823–834.
  46. Zhu, N.; Zhu, Y.; Yu, N.; Wei, Y.; Zhang, J.; Hou, Y.; Sun, A.-d. Evaluation of microbial, physicochemical parameters and flavor of blueberry juice after microchip-pulsed electric field. Food Chem. 2019, 274, 146–155.
  47. Mannozzi, C.; Rompoonpol, K.; Fauster, T.; Tylewicz, U.; Romani, S.; Dalla Rosa, M.; Jaeger, H. Influence of pulsed electric field and ohmic heating pretreatments on enzyme and antioxidant activity of fruit and vegetable juices. Foods 2019, 8, 247.
  48. Schottroff, F.; Gratz, M.; Krottenthaler, A.; Johnson, N.B.; Bedard, M.F.; Jaeger, H. Pulsed electric field preservation of liquid whey protein formulations–influence of process parameters, pH, and protein content on the inactivation of Listeria innocua and the retention of bioactive ingredients. J. Food Eng. 2019, 243, 142–152.
  49. Xu, S.; Walkling-Ribeiro, M.; Griffiths, M.; Corredig, M. Pulsed electric field processing preserves the antiproliferative activity of the milk fat globule membrane on colon carcinoma cells. J. Dairy Sci. 2015, 98, 2867–2874.
  50. Zhao, W.; Yang, R. Pulsed electric field induced aggregation of food proteins: Ovalbumin and bovine serum albumin. Food Bioprocess Technol. 2012, 5, 1706–1714.
  51. Xiang, B.Y.; Ngadi, M.O.; Ochoa-Martinez, L.A.; Simpson, M.V. Pulsed electric field-induced structural modification of whey protein isolate. Food Bioprocess Technol. 2011, 4, 1341–1348.
  52. Sui, Q.; Roginski, H.; Williams, R.P.; Versteeg, C.; Wan, J. Effect of pulsed electric field and thermal treatment on the physicochemical and functional properties of whey protein isolate. Int. Dairy J. 2011, 21, 206–213.
  53. Yu, L.J.; Ngadi, M.; Raghavan, V. Proteolysis of cheese slurry made from pulsed electric field-treated milk. Food Bioprocess Technol. 2012, 5, 47–54.
  54. Banjare, K.; Kumar, M.; Kumar, R.; Kartikyen, S.; Goel, B.; Uprit, S. Perspective role of goat milk and products: A review. Int. J. Chem. Stud. 2017, 5, 1328–1338.
  55. Ahmed, M.; Pickova, J.; Ahmad, T.; Liaquat, M.; Farid, A.; Jahangir, M. Oxidation of lipids in foods. Sarhad J. Agric. 2016, 32, 230–238.
  56. Salari, S.; Jafari, S.M. The influence of ohmic heating on degradation of food bioactive ingredients. Food Eng. Rev. 2020, 12, 191–208.
  57. Ghnimi, S.; Budilarto, E.; Kamal-Eldin, A. The new paradigm for lipid oxidation and insights to microencapsulation of omega-3 fatty acids. Compr. Rev. Food Sci. Food Saf. 2017, 16, 1206–1218.
  58. Deeth, H.; Fitz-Gerald, C. Lipolytic enzymes and hydrolytic rancidity. In Advanced Dairy Chemistry; Springer: Boston, MA, USA, 2006; Volume 2, pp. 481–556.
  59. Shinn, S.E.; Ruan, C.M.; Proctor, A. Strategies for producing and incorporating conjugated linoleic acid–rich oils in foods. Annu. Rev. Food Sci. Technol. 2017, 8, 181–204.
  60. Garcia-Amezquita, L.; Primo-Mora, A.; Barbosa-Cánovas, G.; Sepulveda, D. Effect of nonthermal technologies on the native size distribution of fat globules in bovine cheese-making milk. Innov. Food Sci. Emerg. Technol. 2009, 10, 491–494.
  61. Pataro, G.; Ferrari, G. Limitations of pulsed electric field utilization in food industry. In Pulsed Electric Fields to Obtain Healthier and Sustainable Food for Tomorrow; Elsevier: Amsterdam, The Netherlands, 2020; pp. 283–310.
  62. Ranadheera, C.; Evans, C.; Baines, S.; Balthazar, C.F.; Cruz, A.G.; Esmerino, E.A.; Freitas, M.Q.; Pimentel, T.C.; Wittwer, A.; Naumovski, N. Probiotics in goat milk products: Delivery capacity and ability to improve sensory attributes. Compr. Rev. Food Sci. Food Saf. 2019, 18, 867–882.
  63. Turkmen, N.; Akal, C.; Özer, B. Probiotic dairy-based beverages: A review. J. Funct. Foods 2019, 53, 62–75.
  64. O’mahony, J.A.; Fox, P.F. Milk Proteins: Introduction and Historical Aspects. In Advanced Dairy Chemistry Volume 2 Lipids, 4th ed.; McSweeney, P.L., Fox, P.F., Eds.; Springer Science & Business Media: Boston, MA, USA, 2013; Volume 1A, pp. 43–85.
  65. Duque, S.M.M.; Leong, S.Y.; Agyei, D.; Singh, J.; Larsen, N.; Oey, I. Understanding the impact of Pulsed Electric Fields treatment on the thermal and pasting properties of raw and thermally processed oat flours. Food Res. Int. 2020, 129, 108839.
  66. Zhang, Z.-H.; Zeng, X.-A.; Brennan, C.S.; Brennan, M.; Han, Z.; Xiong, X.-Y. Effects of pulsed electric fields (PEF) on vitamin C and its antioxidant properties. Int. J. Mol. Sci. 2015, 16, 24159–24173.
  67. Buckow, R.; Chandry, P.S.; Ng, S.Y.; McAuley, C.M.; Swanson, B.G. Opportunities and challenges in pulsed electric field processing of dairy products. Int. Dairy J. 2014, 34, 199–212.
  68. Yilmaz, M.; Evrendilek, G.A. Impact of the pulsed electric field treatment on bioactive food compounds: Bioaccessibility and bioavailability. Food Sci. Nutr. 2017, 7, 1–6.
  69. Sharma, P.; Oey, I.; Everett, D.W. Effect of pulsed electric field processing on the functional properties of bovine milk. Trends Food Sci. Technol. 2014, 35, 87–101.
  70. Michalski, M.-C.; Michel, F.; Sainmont, D.; Briard, V. Apparent ζ-potential as a tool to assess mechanical damages to the milk fat globule membrane. Colloids Surf. B Biointerfaces 2002, 23, 23–30.
  71. Riener, J.; Noci, F.; Cronin, D.A.; Morgan, D.J.; Lyng, J.G. Effect of high intensity pulsed electric fields on enzymes and vitamins in bovine raw milk. Int. J. Dairy Technol. 2009, 62, 1–6.
  72. Bendicho, S.; Espachs, A.; Arantegui, J.; Martín, O. Effect of high intensity pulsed electric fields and heat treatments on vitamins of milk. J. Dairy Res. 2002, 69, 113–123.
  73. Noël, L.; Chekri, R.; Millour, S.; Vastel, C.; Kadar, A.; Sirot, V.; Leblanc, J.-C.; Guérin, T. Li, Cr, Mn, Co, Ni, Cu, Zn, Se and Mo levels in foodstuffs from the Second French TDS. Food Chem. 2012, 132, 1502–1513.
  74. Holt, C. Encyclopedia of Dairy Sciences; Fuquay, J.W., Ed.; Academic Press: San Diego, CA, USA, 2011; pp. 917–924.
  75. Poulsen, N.A.; Rybicka, I.; Poulsen, H.D.; Larsen, L.B.; Andersen, K.K.; Larsen, M.K. Seasonal variation in content of riboflavin and major minerals in bulk milk from three Danish dairies. Int. Dairy J. 2015, 42, 6–11.
  76. Zamberlin, Š.; Antunac, N.; Havranek, J.; Samaržija, D. Mineral elements in milk and dairy products. Mljekarstvo Časopis Za Unaprjeđenje Proizv. I Prerade Mlijeka 2012, 62, 111–125.
More
© Text is available under the terms and conditions of the Creative Commons Attribution (CC BY) license; additional terms may apply. By using this site, you agree to the Terms and Conditions and Privacy Policy.
This entry is offline, you can click here to edit this entry!
ScholarVision Creations
Feedback