Accepted Manuscript

Title: Effect of processing on the bioaccessibility of bioactive

compounds – a review focusing on carotenoids, minerals,
ascorbic acid, tocopherols and polyphenols

Authors: Antonio Cilla, Lourdes Bosch, Reyes Barberá,

Amparo Alegrı́a

PII: S0889-1575(17)30026-1
DOI: http://dx.doi.org/doi:10.1016/j.jfca.2017.01.009
Reference: YJFCA 2823

To appear in:

Received date: 17-11-2016

Revised date: 11-1-2017
Accepted date: 18-1-2017

Please cite this article as: Cilla, Antonio., Bosch, Lourdes., Barberá, Reyes., & Alegrı́a,
Amparo., Effect of processing on the bioaccessibility of bioactive compounds – a review
focusing on carotenoids, minerals, ascorbic acid, tocopherols and polyphenols.Journal
of Food Composition and Analysis http://dx.doi.org/10.1016/j.jfca.2017.01.009

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Effect of processing on the bioaccessibility of bioactive compounds – a

review focusing on carotenoids, minerals, ascorbic acid, tocopherols

and polyphenols

Antonio Cilla1, Lourdes Bosch2, Reyes Barberá1 and Amparo Alegría1*

1
Nutrition and Food Science Area. Faculty of Pharmacy. University of Valencia. Avda.

Vicente Andrés Estellés s/n. 46100. Burjassot, Valencia, Spain.

2
Department of Pharmacy, Faculty of Health Sciences, Universidad CEU Cardenal

Herrera. Avda. Seminario s/n. 46113. Moncada, Valencia, Spain.

*Corresponding author: amparo.alegria@uv.es. Tel.: +34963544907. Fax: +343544954.

Type of paper: Study Review

HIGHLIGHTS

 The effect of processing on bioaccessibility of bioactive compounds is reviewed

 Thermal treatment can exert a positive or negative effect
 Emerging technologies as high pressure processing could improve
bioaccessibility
 The effect depends on the food matrix and processing variables
 There is a need of standardization and validation of the in vitro models used

ABSTRACT

Health benefits of bioactive compounds depends not only on the intake levels but also

on their bioavailability (BAv). In vitro methods to simulate gastro-intestinal digestion

allow to determine the bioaccessibility (BAcs) of these compounds, as a first step of

BAv, and can be used to evaluate the effect of processing on them to design functional

foods with increased health-promoting effects. The impact of traditional processing

technologies such as thermal treatment and novel emerging non-thermal technologies

such as high pressure processing, high-intensity pulsed electric fields and ultrasound on

BAcs of bioactive compounds as carotenoids, minerals, ascorbic acid, tocopherols,

polyphenols and total antioxidant capacity is described in this review article. In general,

combination of high pressure processing and thermal treatment in the presence of oil for

carotenoids, thermal treatment for ascorbic acid and polyphenols, and high pressure

processing for minerals, tocopherols and total antioxidant capacity would increase BAcs

of these bioactive compounds. Ultrasound processing is likely to improve polyphenol

compound BAcs and high-intensity pulsed electric fields could improve carotenoid,

vitamin C and phenolic compounds BAcs, as well as antioxidant activity. However,

general guidelines seem not to be applicable for every case study and more research in

this field is needed.

Keywords: Food analysis, food composition, bioaccessibility, bioactive compounds,

thermal treatment, high pressure processing, high-intensity pulsed electric fields,

ultrasound, review.

1. Introduction

The objectives of nutrition are to provide nutrients in such a quantity and quality

to accomplish with body requirements, and bioactive compounds to improve health and

well-being and to prevent the risk to suffer diseases (“optimal” nutrition). The potential

of bioactive components within foods to exert their effects in the body depends on their

matrix release, changes during digestion, uptake, metabolism, and biodistribution, even

before considering dose- and host-related factors; that is, these compounds must be

bioavailable prior to exerting their bioactivity (Bohn et al., 2015).

The term bioavailability (BAv) has several working conditions and there is no

universally accepted definition. From a nutritional point of view, it is defined as the

fraction of ingested component available for utilization in normal physiological

functions. Besides, BAv includes two additional terms: bioaccessibility (BAcs) and

bioactivity (BAct) (Fernández-García et al., 2009).

Bioaccessibility has received two alternative definitions. The first one is the

fraction of a compound that is released from its food matrix in the gastrointestinal tract

and thus becomes available for intestinal absorption. The second definition is more

stringent and much less widely used. It describes BAcs as the fraction of a compound

that is released from its food matrix in the gastrointestinal tract and thus becomes

available for intestinal absorption including absorption/assimilation into the cells of the
intestinal epithelium and, lastly, pre-systemic intestinal and hepatic metabolism

(Cardoso et al., 2015).

The concept of bioactivity includes events linked to how the bioactive

compound has reached systemic circulation and it is transported and reaches the target

tissue interaction with biomolecules metabolism in these tissues, and all the cascade of

physiological effects it generates. In vitro methods can be evaluated BAcs and/or BAct.

(Fernández-García et al., 2009; Cardoso et al., 2015).

Thus, knowledge on BAcs as a first step of BAv is of great interest to ascertain

the nutritional quality of a nutrient or bioactive compound not only in terms of

quantities required to achieve nutritional requirements, but also to fine tuning the

development of functional foods (Fernández-García et al., 2009).

Food processing is one of the main determinants on BAv, because it can have a

positive or negative impact by increasing or decreasing BAcs of nutrients and bioactive

compounds, respectively. In fact, processing techniques are becoming more

sophisticated and diverse in response to the growing demand for quality foods, with

nonthermal processing as a useful tool to extend shelf-life and quality of products as

well as to preserve their nutritional and functional characteristics (Sánchez-Moreno et

al., 2009). So, efforts made during processing to reduce the impact of applied

technology on key components of food would be more valuable if nutritional quality

considers both stability and BAcs.

Based on this scenario, the present review provides an updated standpoint on the

effect of food processing (traditional: heat related thermal treatment, and emerging

technologies: high pressure processing, high-intensity pulsed electric fields, ultrasound

processing) on BAcs (determined by in vitro methods) of bioactive compounds and

their impact to design functional foods with increased health-promoting effects as well

as for scientists and consumers to be aware of these advances, but taking into account
that the effect of food processing is more complex than the positive effect that might be

expected intuitively.

2. Methods for bioaccessibility determination

Data from human intervention studies (in vivo assays) constitute the reference

standard and offer the highest scientific evidence referred to the BAv of a nutrient or

bioactive compound. A number of disadvantages, like limitations in experimental

design, difficulties in data interpretation, high cost of equipment and labor and ethical

constraints, limit the utility of in vivo methods for screening the BAv of bioactive

compounds for large number of differently processed foods (Van Buggenhout et al.,

2010). Some in vivo studies have been carried out in humans focusing on the effect that

processing exerts on bioactive compounds BAv, including for instance those published

by Sánchez-Moreno et al. (2004a, 2004b, 2005), Tydeman et al. (2010), Aschoff et al.

(2015) and Martínez-Huélamo et al. (2015).

On the contrary, in vitro models are cost-effective, reproducible and, in general,

rapid methods that can be used to determine the effects of food matrix, processing

methods, and dietary components on BAcs of bioactive compounds in foods and are

used as surrogates for predictive purposes. Nevertheless, despite their potential and

broad applicability, they do not fully mimic the overall processes occurring in vivo,

particularly hormonal and nervous control, feedback mechanisms, mucosal cell activity,

complexity of peristaltic movements and involvement of the local immune system

(Guerra et al., 2012). In addition, there is urgent need for standardization of the in vitro

methods to afford improved study designs more similar to the in vivo situation and for

allowing comparisons of results among laboratories (Hur et al., 2011; Cilla et al., 2013;
Ting et al, 2015). A harmonized static in vitro digestion model has been proposed by

Minekus et al. with consensus protocol, within the COST FA1005 Action INFOGEST

Network, based on human gastrointestinal physiologically relevant conditions (Minekus

et al., 2014).

For each nutrient or bioactive compound, specific in vitro digestion models must

be tailored, changed, tested, and validated against in vivo studies before any conclusion

is drawn from their results. It can therefore not be taken for granted that an in vitro

assay will yield results applicable to the in vivo situation. Therefore, whenever possible,

in vivo studies should be used for the validation of in vitro models (Cardoso et al.,

2015).

In vitro methods have been developed to simulate the physiological conditions

and the sequence of events that occur during digestion in the human gastrointestinal

tract. In a first step, simulated gastrointestinal digestion (gastric and intestinal stages,

and in some cases a salivary stage) is applied to homogenized foods or isolated

bioactive compounds in a closed system, with determination of the soluble component

fraction obtained by centrifugation (solubility method) or dialysis of soluble

components across a semi-permeable membrane (dialysis method) to obtain the

bioaccessible fraction. Simulated gastrointestinal digestion can be performed with static

models where the products of digestion remain largely immobile and do not mimic

physical processes such as shear, mixing, hydration (Alegría et al., 2015). Dynamic

models can also be used, with gradual modifications in pH and enzymes, and removal

of the dialyzed components – thereby better simulating the actual in vivo situation.

In vitro bioavailability assays based on in vitro digestion and on measurements

of the soluble or dialyzable bioactive compounds have been improved by incorporating

cell cultures. The Caco-2 cell model is the most widely used and validated intestinal
epithelium cell model. Although colonic in origin, Caco-2 cells undergo spontaneous

differentiation in cell culture to form a monolayer of well polarized cells at confluence,

showing many of the functional and morphological properties of mature human

enterocytes (Pinto et al., 1983).

All these systems evaluate the aforementioned term BAcs, and can be used to establish

trends in relative BAv (Parada  Aguilera, 2007; Etcheverry et al., 2012, Guerra et al.,

2012; Cardoso et al., 2015).

3. Brief overview of food processing

Processing of food products may have positive or negative effects on BAcs of

bioactive compounds as do the environmental conditions during its passage along the

gastrointestinal tract (Wang  Bohn, 2012). The impact of food processing on the BAcs

of bioactive compounds has been indicated not to be a simple cause-effect relationship

(Watzke, 1998; Van Buggenhout et al., 2010). In this review, the focus lies on

traditional processing technologies such as heat related thermal treatment (TT) and on

novel emerging nonthermal techniques such as high pressure processing (HPP), high-

intensity pulsed electric fields (HIPEF) and ultrasound treatment, that are used to

preserve manufactured food or cooking, as well as during the food preparation, i.e. pre-

treatments on raw material to obtain food.

TT is an important food preservation unit operation which includes, among

others, pasteurization and sterilization by methods such as steaming, boiling, roasting,

microwave and so on. It can be applied to either solid or liquid products: to prepare the

product (i.e. cooking the product for added BAcs of nutrients), to develop desired

flavours, aroma and color components (i.e. Maillard reaction), modify the food

structure, or to preserve or sterilize the food by heat induced inactivation of

microorganisms, toxins and enzymes (Wang  Bohn, 2012). However, despite the

increase in food safety achieved with TT, there is often a loss in BAcs and/or BAct of

native bioactive compounds and nutrients which are essential for the human diet.

As compared to TT at atmospheric pressure that would not be always uniform,

HPP exerts a pressure at a given position and time which is the same in all directions,

transmitted uniformly, immediately through the pressure transferring medium and

independent of product size and geometry (Oey et al., 2008). This methodology can be

applied to treat (sterilization) of products which are highly prone to spoilage and

damage through processing (i.e. fresh dairy products, meat, seafood, fruits and

vegetables). The major benefits include: no formation of unwanted compounds

(products from heat treatment), best preservation of natural nutritional value, flavor,

appearance, texture, and possible production of safe food (microbial safety) with the

highest quality (nutritional and sensorial). On the contrary, high process costs

(investment and maintenance) and the fact that available equipment is mostly for batch

processes and not industrial scale, are its main limitations (Wang  Bohn, 2012).

HIPEF technology involves the application of very short pulses (1–10 μs) of

high electric intensity (10–100 kV/cm) to treat food products placed between two

electrodes without substantially heating the product. This technique is an alternative to

thermal treatment to preserve food against microorganisms that has been used in order

not to compromise nutritional and sensorial features (Zhang et al., 2015). Another

nonthermal technique such as ultrasound has been used to induce changes in structural

and rheological properties of different food matrices or to extract compounds in them

(Anese et al., 2013); but there are only a few studies addressing the effect of these

nonthermal processes on BAcs of bioactive compounds.

Pressure Change Technology (PCT) is a recent gentle treatment that uses an

inert gas, such as nitrogen, helium or argon, at pressure up to 50 MPa and temperature
between 0 and 40ºC and can be applied to foodstuff to inactivate microorganisms

(Bönsch et al., 2007). It improves quality parameters in food if compared with other

technologies. In fact, it may also improve BAcs of bioactive compounds, like

carotenoids, due to a decrease in particle size, but further studies are needed (Aschoff et

al., 2016).

4. Impact of food processing on bioaccessibility of bioactive compounds

It has been reported that in some cases the total amount of a nutrient may

decrease in the food chain due to chemical degradation during storage and physical

losses in processing, whereas at the same time the BAcs (and consequently BAv) may

increase most likely as a result of disruption of the cell walls of plant tissues,

dissociation of the nutrient-matrix complexes, or transformation into more active

molecular structures (Parada  Aguilera, 2007). In vitro studies have been performed to

study the impact of several processing methods applied in food manufacturing,

preservation or cooking on the BAcs of bioactive compounds. In general, the effect of

food processing on the BAcs of bioactive compounds is dependent of different factors:

(i) type of processing (ii) type of bioactive compound considered (iii) food matrix

composition and structure and (iv) presence of components that affect absorption

efficiency (Van Buggenhout et al., 2010). It must be taken into account that BACs of

some bioactive compounds such as carotenoid can be also affected by their physical

form of deposition in plant chromoplasts, as it can have an effect on their liberation

efficiency from the food matrices (Schweiggert et al., 2012, 2015).

4.1. Food preparation or pre-treatment / manufacturing

 Recent studies have investigated the effect of several steps (mechanical and

thermal combined treatments) in manufacturing process to obtain food (i.e. extrusion,

roasting, grinding and alkalization treatment and ultrasound processing) on BAcs of

functional ingredient. The effect of extrusion on lycopene BAcs of tomato pulp added

corn extrudates have been studied. Tomato pulp is added as a functional ingredient to

corn grit extrudates (ready-to-eat starch-rich products in different sizes, shapes or

textures that are obtained by using a screw extruder and heating, 130 and 160ºC last

zone treatments). BAcs (solubility and dialysis methods), according to the consensus

method of Minekus et al., 2014) of lycopene was higher after the extrusion process

(higher values at 160ºC), probably because of a release; although the total phenolic

content, total antioxidant activity (DPPH assay) and lycopene content decreased after

the process due to decomposition (showing higher values at 160ºC). The increase in the

extrusion temperature may cause a higher extractability of phenolic compounds and

lycopene and result in more Maillard products leading to higher antioxidant activity

(Tonyali et al., 2016).

Another pre-treatment is the one used in the cocoa powder production from

cocoa beans which includes roasting, grinding and an alkalization (at 80-100ºC for 10-

12 h up to pH 8.2) to darken the color and milden the flavour. Roasting and alkalization

reduced polyphenols content and total antioxidant activity (DPPH and ORAC assays) as

well as BAcs of procyanidins (solubility assay), whereas grinding had the opposite

effect because of the breakdown of the cell walls (Gültekin-Özgüven et al. 2016).

Ultrasound treatment can be used at industrial level to induce changes in the

structure of food like texture and viscosity and can affect the antioxidant properties.

There are few studies addressing the effect of this process on BAcs of bioactive

compounds in some food and by-products, such as lycopene in tomato pulp (Anese et

al., 2013, 2015), polyphenols in olive leaf (Ahmad- Qasem et al., 2014) and phenolic
compounds, total antioxidant capacity and vitamin C in cashew apple bagasee puree

(Fonteles et al., 2016), with discrepancies in their results.

Ultrasound processing does not change the total lycopene concentration of

tomato pulp (Anese et al., 2013, 2015), but cause a marked decrease in its BAcs

(solubility method), probably due to the formation of a stronger network that entrapped

lycopene in the matrix (Anese et al., 2013). However, no differences were reported in

lycopene BAcs in the same processed matrix using a different method (adding an oil-in-

water emulsion just before the in vitro digestion, together with a lipase containing

solution) (Anese et al., 2015). Ultrasound treatment has been also used to extract

phenolic compounds in dried olive leaves, providing a polyphenol content similar to

that of the conventional procedure, but ensuring a better BAcs (Ahmed-Qasem et al.,

2014). However, when this process was applied on cashew apple bagasse, it increased

antioxidant activity, total phenolic compounds and vitamin C content, as well as total

phenolic compounds BAcs, but decreased slightly vitamin C BAcs (Fonteles et al.,

2016).

4.2. Food processing: preservation treatment and cooking

In the past 15 years, in vitro studies have been performed to study the impact of

TT, HPP or the combination of both processing methods, as well as HIPEF, on the

BAcs of bioactive compounds, mostly in carotenoids (Table 1), but also in minerals

(Table 2) and antioxidants (including total antioxidant capacity, tocopherols, ascorbic

acid and polyphenols) (Table 3).

4.2.1 Thermal processing

 To evaluate carotenoids BAcs (Table 1), solubility is the main method used,

followed by dialysis or combined systems solubility / Caco-2 cells. It is important to

highlight that Veda et al. (2006) indicate that equilibrium dialysis is not suitable for the

determination of β-carotene BAcs, due to lower micellar fraction obtained containing

the bioaccessible β-carotene after digestion.

In general, heat processing (cooking and other processing methods) increase the

BAcs of carotenoids by destroying the integrity of cell wall and membranes of

organelles in which carotenoids are located, so that the digestive enzymes may work

more efficiently for releasing of carotenoids from food matrix into oil droplets (Hedrén

et al., 2002; Thakkar et al., 2009; Colle et al., 2010a; Lemmens et al., 2010; Page et al.,

2012; Kamiloglu et al., 2014). It has also been suggested that heat denatures protein-

carotenoid complexes that limit carotenoid BAcs, favoring their release from food

matrix (Colle et al., 2010a; Lemmens et al., 2010). Moreover, the structural

characteristics of the starting material are very important for the increase in carotenoids

BAcs. In fact, mechanical disruption or homogenization prior to the heat treatment can

enlarge the surface area for the digestive enzymes attack, and thereby is an

advantageous approach for releasing carotenoids from the food matrix (Hedrén et al.,

2002; Bengtsson et al., 2010). In addition, the positive impact of heating is potentiated

by the presence of dietary fat (oil), facilitating an hydrophobic environment for the

released carotenoids (Hedrén et al., 2002; Veda et al., 2006; Hornero-Méndez 

Mínguez-Mosquera, 2007; Tumuhinbise et al., 2009; Knockaert et al., 2012b; Victoria-

Campos et al., 2013). On the contrary, Anese et al. (2015) reported a decrease in

lycopene BAcs with the increase of oil content added. Accordingly, and considering the

abovementioned premises, an increase in carotenoids BAcs due to heat processing has

been observed in different foods such as carrots (Hedrén et al., 2002; Hornero-Méndez

 Mínguez-Mosquera, 2007; Lemmens et al., 2009, 2010, 2011; Knockaert et al.,

2011), tomatoes (Tibäck et al., 2009; Svelander et al., 2010; Colle et al., 2010a, 2013;

Gupta et al., 2011; Page et al., 2012; Kamiloglu et al., 2014), cassava (Thakkar et al.,

2009; Failla et al., 2012), peppers (Victoria-Campos et al., 2013; Pugliese et al., 2013,

2014), orange sweet potatoes (Tumuhimbise et al., 2009; Bengtsson et al., 2010) and

pumpkin, amaranth leaves, fenugreek leaves (Veda et al., 2006), green beans

(McInerney et al., 2007), kale and spinach (O’Sullivan et al., 2010), orange juice

(Aschoff et al., 2015) and lutein in mloukhiya (a Tunisian dish with dried jute leaves,

tomato paste, olive oil) (Dhuique-Mayer et al., 2016). Carotenoid BAcs was poor in

cooked provitamin A-biofortified pumpkin, lower for pulp boiled in water with sugar

compared to water alone or steamed; but it was observed an improvement with the time

of cooking (Ribeiro et al., 2015).

On the contrary, TT can reduce carotenoids BAcs inducing structural changes in

carotenoids, mainly isomerization, altering their solubility and consequently their

micellarization (Victoria-Campos et al., 2013). Besides, xanthophylls seem to be more

affected than carotenes (Gupta et al., 2011; Pugliese et al., 2014). The conflicting

information on carotenoid BAcs can be due to different factors, such as differences in

methodologies to determine BAcs and (slightly) different starting material (i.e. particle

size, presence/absence of oil during processing or during digestion) (Lemmens et al.,

2011; Palmero et al., 2014). In this regard, TT has evoked carotenoid BAcs decrease in

different foods such as courgettes, peppers and tomatoes (Ryan et al., 2008), broccoli

and savoy cabbage (O’Sullivan et al., 2010) tomato juice (Gupta et al., 2011), milk-

based fruit beverages (Cilla et al., 2012; Rodríguez-Roque et al., 2016) and other fruit

juice-based beverages (Rodríguez-Roque et al., 2016), orange juice (Victoria-Campos et

al., 2013), red chili peppers (Pugliese et al., 2014) and -carotene in mloukhiya

(Dhuique-Mayer et al., 2016); and no changes were observed in kiwifruit puree

(Benlloch-Tinoco et al., 2015) and plum and cabbage (Kaulmann et al., 2016).
 Concerning minerals (Table 2) it is believed that BAcs can be enhanced as a

result of TT due to the softening of the food matrix and consequently the release of

protein-bound minerals (Hemalatha et al., 2007), and/or by modifying the contents of

solubility inhibitors such as oxalates, phytates, tannins and phenolic compounds (Viadel

et al., 2006a). This fact has been observed for Ca cell uptake in ready-to-eat lentils

(Viadel et al., 2006a) and for Fe and Zn BAcs in cereals and pulses treated by pressure-

cooking, microwave and cooked in a regular pan (boiled) (Hemalatha et al., 2007,

Pereira et al., 2016). On the other hand, most of the studies reviewed showed a decrease

in Ca, Fe, Zn or Se BAcs in foods such as pulses and cereals (Sebastiá et al., 2001;

Viadel et al., 2006a, 2006b; Hemalatha et al., 2007; Khanam  Platel, 2016), green

leafy vegetables (Khanam  Platel, 2016) and infant formulas (Galán  Drago, 2014),

or P BAcs in milk-based fruit beverages (Cilla et al., 2011). This situation can be

ascribed to the degradation of ascorbic acid (which is an enhancer of Fe absorption), the

generation of negative mineral-nutrient interactions and changes in chemical structure

due to oxidation of the element (Galán  Drago, 2014; Khanam  Platel, 2016).

Another possibility can be that in the cooking process of legumes, the soaking and

cooking water is drained off and consequently the mineral BAcs is reduced (Sebastiá et

al., 2001). In fact, it has been reported that soaking legumes previous to TT can exert a

negative effect on Fe and Zn BAcs, probably because of a mineral lost in the water

(Pereira et al., 2016).

Regarding the effect of TT on antioxidants BAcs (Table 3) conflicting results

have been reported. A positive effect has been indicated for ascorbic acid in milk-based

fruit beverages (Cilla et al., 2012) and for total phenolics and antioxidant capacity in

grape and orange juice (He et al., 2016) treated by TT vs. HPP; however, recent studies

reported a decrease in vitamin C and antioxidant capacity in similar beverages treated

by TT vs. HPP (Rodríguez-Roque et al., 2015, 2016). Moreover, a positive effect of TT

has been observed in green tea catechins boiling-brewed (epigallocatechin,

epigallocatechingallate and epicatechingallate) vs. ambient temperature-brewed (Xie et

al., 2013), probably due to the degradation of complexes formed with proteins; in total

polyphenols from cereal (millet, shorgum, wheat), legumes (green gram and chickpea)

and red beetroot jam (Hithamani  Srinivasan, 2014a, 2014b; Guldiken et al., 2016); in

total flavonoids from dried red beetroot and jam (Guldiken et al., 2016); and total

antioxidant capacity in black carrot jam and marmalade and red beetroot jam

(Kamiloglu et al., 2015; Guldiken et al., 2016). Nevertheless, a negative effect has been

observed for tocopherols in milk-based fruit beverages treated by TT vs. HPP (Cilla et

al., 2012), possibly by the chemical degradation of these compounds due to the heat

treatment, and in mloukhiya (Dhuique-Mayer et al., 2016), because it was poorly

micellarized in the in vitro digestion remaining in oil. A negative effect for total

phenolics and total antioxidant capacity in pickled red beetroot and plum and cabbage

(Guldiken et al., 2016; Kaulmann et al., 2016) has been also reported.

4.2.2. High pressure processing or combined methods

In general, HPP and/or the combination of an initial homogenization step carried

out with HPP followed by TT are expected to exert an increase in bioactive compounds

BAcs. However, depending on the HPP and HPP + TT conditions applied, a decrease in

carotenoids, minerals and antioxidants BAcs has also been reported.

An improvement of carotenoids BAcs has been found in green beans, carrots,

tomato, soya milk-based and fruit juice-based beverages (McInerney et al., 2007;

Knockaert et al., 2011, 2012a, 2012b; Gupta et al., 2011; Svelander et al., 2011; Cilla et

al., 2012; Colle et al., 2013; Rodríguez-Roque et al., 2016) (Table 1) due to HPP and

HPP + TT. The positive effect of HPP has been related to a disintegration of cell

clusters and disruption of cells containing carotenoids (Svelander et al., 2011; Cilla et
al., 2012). The beneficial effect of subsequent TT following HPP, in turn, has been

ascribed to the fact that high temperature can weaken the physical barriers that enclose

carotenoids (since cell membranes are easily destroyed) and to the acceleration of β-

elimination of pectin which results in softening the cell walls. Besides, the addition of

oil to this combined treatment can further increase carotenoid BAcs (Knockaert et al.,

2012b; Colle et al., 2013). On the contrary, a decrease in the BAcs of carotenoids has

been noted in broccoli, tomatoes and fruit juice-based beverages (McInerney et al.,

2007; Colle et al., 2010b; Knockaert et al., 2012a; Rodríguez-Roque et al., 2016). This

negative effect has been linked with some vegetables that have firmer cellular structures

that need higher processing pressure (broccoli vs. green beans) (McInerney et al., 2007)

and to the changes in pulp microstructure that can form a fiber network which entraps

lycopene making it less accessible to the digestive enzymes and bile salts (Colle et al.,

2010b). In order to study the role of the soluble/insoluble phases and the natural

structural barriers in determining the carotenoid BAcs upon HPP, tomato-based model

systems with (chromoplast and cell cluster fractions) and without (model systems

containing the carotenoid enriched oil fraction) structural barriers have been developed.

In presence of structural barriers, HPP affect them with an enhanced carotenoid release

and improvement of BAcs. In the absence of these barriers, the soluble and insoluble

phases determined the carotenoid BAcs (Palmero et al., 2016).

In the case of minerals (Table 2) conflicting results were found again. An

increase in Ca and P solubility and P bioavailability in Caco-2 cells has been reported in

milk-based fruit beverages due to HPP vs. TT. In this regard, HPP at >300 MPa can

exert a disruptive effect on the ionic and hydrophobic interactions of calcium phosphate

with casein micelles, resulting in increases of soluble Ca and P levels (Cilla et al.,

2011). Similarly, increases in Ca, Fe and/or Zn has been observed in apples and

algarrobo seeds, attributed to the changes induced in the food matrix such as disruption
of plant cell walls (Briones-Labarca et al., 2011a, 2011b). However, these same authors

found decreases in these minerals in the same foods with different digestion methods

(solubility and dialysis) and at different treatment times, but without giving a plausible

explanation.

Finally, regarding the effect of HPP on antioxidants BAcs (Table 3) an increase

in total antioxidant capacity (DPPH method) in apples and algarrobo seeds has been

described due to the release of antioxidants into the extracellular environment after the

disruption of cell walls (Briones-Labarca et al., 2011a, 2011b). Likewise, an increase in

tocopherols BAcs has been reported in milk-based fruit beverages. It is possible that

HPP affects the food matrix: modifying the location of tocopherols in the food or

altering the amounts of absorption effectors (fibers, fat and phytosterols), thereby

making these compounds more bioaccessible (Cilla et al., 2012). In the same way, an

increase in vitamin C and antioxidant capacity due to HPP vs. TT in fruit juice, milk-

based fruit beverages and soy milk-based fruit beverages has been reported (Rodríguez-

Roque et al., 2015, 2016). On the opposite side, a decrease in ascorbic acid in milk-

based fruit beverages has been quoted, probably due to the formation of aggregates

originated by HPP, leading to an increase in viscosity and making the ascorbic acid

molecules less accessible during digestion (Cilla et al., 2012) and a decrease in total

phenolics BAcs in high pressure homogenization processing applied to apple juice,

probably due to degradation of the polyphenols by polyphenol oxidase which contact is

promoted with the treatment, despite the rupture in the cellular structure caused by it.

However, this effect was not observed in orange and grape juice (He et al., 2016).

4.2.3. High-intensity pulsed electric fields (HIPEF)

To our knowledge, there are only two studies in terms of the effect of HIPEF

(high-intensity pulsed electric fields) technique on BAcs of bioactive compounds in

fruit juice-based beverages (water-, milk-, or soymilk-fruit juice beverages) (Rodríguez-

Roque et al., 2015, 2016). In general, a decrease of carotenoids BAcs has been found

with HIPEF treatment, but, in some cases, improved it in comparison with untreated

beverages (Table 1). The treatment did not change or improved the BAcs of vitamin C

and phenolic compounds. Regarding the effect on antioxidant activity, HIPEF improved

it vs. untreatment in milk-fruit juice and, in general, values obtained were better vs. TT

(Table 3).

5. Conclusions

In this review we have found strengths and weaknesses related the topic of the

effect of processing on BAcs of bioactive compounds.

The strong points rely on: (i) better knowledge in processing technology with

improved efficiency to increase BAcs in foods, and (ii) the production of foodstuffs

with enhanced BAcs of bioactive compounds (functional foods) can improve the

consumer’s perception of the product, subsequently increasing the profitability of it, and

thus potentiating its inclusion in the dietary lifestyle of population.

On the other hand, limitations are: (i) processing variables (i.e temperature,

particle size, pressure applied) need to be carefully controlled so that data can be scaled

up in to industrial applications, and (ii) the need of using a consensus standard method

to evaluate the biological effects of bioactive compounds in order to compare possible

data obtained.

As a final remark, emerging technologies could improve the BAcs of bioactive

compounds. It can be stated that, in general and based on the results reported so far,

combination of HPP + TT in the presence of oil for carotenoids, TT for ascorbic acid

and polyphenols, and HPP for minerals, tocopherols and total antioxidant capacity,
would be the best options to increase BAcs of these bioactive compounds maximizing

their potential health benefits. Ultrasound processing is likely to improve polyphenol

compound BAcs and HIPEF could improve carotenoid, vitamin C and phenolic

compounds BAcs, as well as antioxidant activity.

However, general guidelines on the use of food processing (TT and/or HPP,

HIPEF and ultrasound) to improve bioactive compounds BAcs seem not to be

applicable for every case study and more research in this field is needed.

Funding

This research did not receive any specific grant from funding agencies in the public,

commercial, or not-for-profit sectors.

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http://www.intechopen.com/books/nutrition-well-being-and-health/health-promoting-

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foods. Trends in Food Science and Technology, 9, 320-327.

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 Table 1. Effect of processing on measured bioaccessibility of carotenoids.

Bioactive In vitro digestion Effect on

Food Processing Reference
compound method bioaccessibility

TT: Cooking, dietary fat and

Cooking and oil mechanical disruption

α- and β- Hedrén et al.,
Carrots cooking (20-40 Solubility (pulping) ↑ α- and β-
carotene 2002
and 60% oil carotene release from

addition) food matrix

TT: Dialysis not suitable for

Carrot
lipid-soluble components
Pressure-cooking
Pumpkin
(15 psi/10min) Solubility and Both methods ↑ β-
β- carotene Veda et al., 2006
Amaranth and dialysis carotene (Stir-frying >>
Stir-frying
Fenugreek pressure-cooking)
(100ºC/10min)
leaves
with oil (9%)

TT:

Cooking
Hornero-Méndez
α- and β- (100ºC/15min) Cooking and oil addition
Carrots Solubility and Mínguez-
carotene and oil cooking ↑ α- and β- carotene
Mosquera, 2007
(5-10% oil
addition)

No effect on carrots
HPP:
α- and β- Carrots, green ↑ lutein at 600 MPa in
400 or 600 McInerney et al.,
carotene and beans and Solubility green beans
MPa/ambient 2007
lutein broccoli
↓ β-carotene at 400 and
Tª/2min
600 MPa in broccoli

↑ β- carotene with all

four methods

↓ Lycopene in red pepers

TT: and courgettes (no effect

β- carotene, Courgette in tomatoes)

Boling (10min)
Solubility/Caco-2
lycopene, β- (zucchini), red ↓ β-cryptoxanthin in all
Grilling (10min) cells (only lutein Ryan et al., 2008
cryptoxanthin pepper and samples and all methods
from courgettes)
and lutein tomato Microwave (50s)
↓ Lutein in red peppers
Steaming (10min)
and tomatoes, no effect in
courgettes and ↑ cell
uptake only with
microwave
HPP: High pressure processing. TT: Thermal treatment.
Solubility method: Determination of the soluble component fraction obtained by centrifugation to obtain the
bioaccessible fraction

Dialysis method: Dialysis of soluble components across a semi-permeable membrane to obtain the bioaccessible
fraction
 Table 1. (continued-I)

Bioactive In vitro digestion Effect on

Food Processing Reference
compound method bioaccessibility

TT:

Boiling
(95ºC/30min)

Gari: β-carotene isomers were

β-carotene
Fermentation + not different in boiling
isomers (all- Thakkar et al.,
Cassava roasting Solubility and gari processing but
trans, 13-cis 2009
(165ºC/10min) decreased with fufu
and 9-cis)
processing
Fufu:
Fermentation +
boiling
(100ºC/10min)

TT:

Boiling
(92ºC/20min)
↑ β- carotene with all
Steaming
Orange fleshed four methods (frying > Tumuhimbise et
β-carotene (94ºC/30min) Solubility
sweet potatoe boiling = steaming > al., 2009
Frying baking)
(170ºC/10min)

Baking
(180ºC/15min)

TT:
↑ lycopene when intense
Heat shock
Crushed crushing followed by Tibäck et al.,
Lycopene (95ºC/8min) Solubility
tomatoes heat treatment (heat 2009
Boiling shock or boiling)
(100ºC/20min)

TT:
↑ β-carotene with
90ºC (0-50min) Lemmens et al.,
β-carotene Carrots Solubility increasing temperature
100ºC (0-20min) 2009
and time
110ºC (0-14min)

TT:

All-E- β- Boiling (3 and 25 ↑ All-E- β-carotene in Lemmens et al.,

Carrots Solubility
carotene min) boiling (25min) 2010
 Table 1. (continued-II)

Bioactive In vitro digestion Effect on

Food Processing Reference
compound method bioaccessibility

TT:
Heat treatments ↑ β-
Boiling
carotene and lycopene
(100ºC/20min)
(LTLT and HTST
LTLT
preferable)
β-carotene Svelander et al.,
Tomato (60ºC/40min) Solubility
and lycopene Additional boiling no 2010
LTLT+boiling
further improve BAcs
HTST
Lycopene more stable
(90ºC/10min)
than β-carotene
HTST+boiling

TT:
β-carotene ↑ all-trans and 13-cis in
BOL
isomers Orange fleshed the order: Bengtsson et al.,
HOM Solubility
(all-trans sweet potato HOM = POB > POA > 2010
POA
and 13-cis) BOL
POB

Lycopene
TT:
isomers ↑ all-trans and cis- only Colle et al.,
Tomato pulp (60ºC to Solubility
(all-trans and at 130ºC and 140ºC 2010a
140ºC/30min)
cis-)

HPP and HPP+TT ↓

HPP (84-1327bar)
lycopene Colle et al.,
Lycopene Tomato pulp followed or not by Solubility
BAcs ↓ up to 479bar, 2010b
TT (90ºC/30min)
then remained constant

↓ 9-cis in broccoli and

savoy cabbage and ↑ in
Broccoli
β-carotene spinach
Boiling
isomers (all- Kale O’Sullivan et al.,
(10min, except Solubility ↓ 13-cis in broccoli and ↑
trans, 13-cis Spinach 2010
spinach 3min) in spinach
and 9-cis)
Savoy cabbage
↓ all-trans in savoy
cabbage and ↑ in kale
BOL: boiling followed by pureeing and oil addition. HOM: homogenization followed boiling and oil addition. LTLT:
low temperature, low time. HTST: high temperature, short time. POA: cooking following oil addition. POB: oil
addition to flour followed by cooking.
 Table 1. (continued-III)

In vitro
Bioactive Effect on
Food Processing digestion Reference
compound bioaccessibility
method

Slight ↑ at 90ºC and

TT: 100ºC/0-60min
Lemmens et al.,
β-carotene Carrot (90ºC to 120ºC/0- Solubility Large ↑ 110ºC and
2011
60min) 120ºC even at 5 and
10min

TT- equivalent
thermal HPP
↑ β-carotene by all
70ºC/2min-
treatments (TT and
500MPa/25ºC/16min Knockaert et al.,
β-carotene Carrot Solubility equivalent thermal HPP)
90ºC/10min- 2011
except HPP
600MPa/45ºC/20min
(600MPa/117ΊC/9.6min)
121.1ºC/3min-
600MPa/117ºC/9.6min

TT combined or not
with HPP: ↑ β-carotene with HPP,
PATP and TP (in raw
HPP
juice) and ↑ with HPP,
(700MPa/30ºC/5min)
Tomato juice preheat and TP (in hot
Preheat break juice)
β-carotene Raw or hot- Gupta et al.,
Solubility
and lycopene (0.1MPa/65ºC/5min) ↓ lycopene with preheat 2011
break
(93ºC/60s) PATP (in raw juice) and ↑ with
all treatments (in hot
(700MPa/100ºC/5min)
break juice)
TP

(0.1MPa/100ºC/5min)

β-carotene
TT:
(all-trans
Only ↑ all-trans α- and
and 13-cis), (90-95ºC/40min)
β-carotene in carrot
α-carotene Carrot, tomato Solubility/Caco-2
TT+HPP: emulsions and ↑ α- and Svelander et al.,
(all-trans) and mixed cells (only β-
10MPa/1 cycle β-carotene in mixed 2011
and lycopene emulsions carotene)
emulsions for HPP+TT
(all-trans, 5- 10MPa/10 cycles
vs. TT
cis and cis-
100MPa/1 cycle
group)
PATP: pressure-assisted thermally processed. TP: thermally processed.
 Table 1. (continued-IV)

In vitro
Bioactive Effect on
Food Processing digestion Reference
compound bioaccessibility
method

Neoxantin+9-
In general ↓ all
cis
carotenoids with HPP
violaxanthin, TT
Milk-based and TT vs. untreated,
zeaxanthin,
(whole milk, (90ºC/30 s) except ↑ all carotenoids
lutein,
skimmed milk Solubility with HPP for soya-milk Cilla et al., 2012
HPP
zeinoxanthin,
and soya milk) samples
β- (400 MPa/40ºC/ 5
fruit beverages HPP confers↑ BAcs
cryptoxanthin min)
carotenoids (better in
and β-
more lipophilic) than TT
carotene

Microwave heating: Rapid temperature rise

(hot-break) leads to ↑
Cold-break (25ºC-
Lycopene Tomato puree Diffusion to oil BAacs than slow Page et al., 2012
70ºC)
temperature rise (cold-
Hot-break (90ºC) break)

TT- equivalent
thermal HPP: The positive effect of
HPP in starting tomato
Starting tomato 60ºC/1min-
puree counterbalanced
puree 450MPa/20ºC/15min Knockaert et al.,
Lycopene Solubility by negative effect of
(10MPa and 90ºC/10min- 2012a
subsequent intense
5% oil) 600MPa/45ºC/20min
thermal processes
121ºC/1.5 or 3min- (>117ºC)
600MPa/117ºC

Homogenized + TT Homogenization ↑ BAcs

Carrot puree (90ºC/10min) only ≥ 50 MPa

with/without oil (5%) Homogenization+TT ↑ Knockaert et al.,

Homogenized
β-carotene Solubility
by HPP (10, 50 Homogenized + HPP BAcs with/without oil 2012b

or 100 MPa) (600MPa/45ºC/20min) Homogenization+HPP

with/without oil (5%) not improves BAcs

Boiling (95ºC/30min) ↑ solubility in boiled and

fufu vs. gari
Cassava and Gari: Fermentation +
transgenic roasting Solubility/ Caco- ↑ cell uptake in boiled Failla et al.,
β-carotene
cassava rich in (165ºC/10min) 2 cells and fufu vs. gari 2012
β-carotene
Fufu: Fermentation + ↑ solubility and cell
boiling (100ºC/10min) uptake in transgenic
 Table 1. (continued-V)

In vitro
Bioactive Effect on
Food Processing digestion Reference
compound bioaccessibility
method

α- and β- Industrial extraction ↑

Orange juice
carotene, β- TT: BAcs carotenoids, but
industrially Stinco et al.,
cryptoxanthin, Pasteurization Solubility pasteurization reduced
processed or 2012
zeaxanthin and (99ºC/15s) them to the level of
hand-squeezed
lutein hand-squeezed

Boiling and grilling ↓

TT:
Free most free and esterified
carotenoids Red and green Boiled carotenoids. Fat ↑ less Victoria-
and mono- and pungent (94ºC/12.5min) Solubility polar carotenoids Campos et al.,
diesterified peppers Grilled (carotenes, β- 2013
xanthophylls cryptoxanthin and
(210ºC/13.2min)
esterified xanthophylls)

TT.
HPP alone no effect
Tomato pulp Microwave
TT alone (↑ BAcs only
with/without
(70ºC, 90ºC, at 120ºC) Colle et al.,
Lycopene oil (coconut, Solubility
120ºC/20min) 2013
olive or fish) HPP followed by TT
5% HPP: higher ↑ BAcs at all
temperatures with oil
100bar/37ºC

↑ BAcs following
Capsanthin,
processing that
zeaxanthin,
compensates the lower
antheraxanthin, Yellow, red
TT: contents in total Pugliese et al.,
violaxanthin, and orange Solubility
Boiling (10min) carotenoids in processed 2013
lutein, β- chili peppers
peppers. Evident for β-
carotene and β-
carotene, zeaxanthin
cryptoxanthin
and β-cryptoxanthin

Capsanthin,
↓ capsanthin, zeaxanthin
zeaxanthin,
and violaxanthin
antheraxanthin,
Red chili TT: Pugliese et al.,
↑ β-carotene (in several
violaxanthin, Solubility
peppers Boiling (10min) cultivars) and β- 2014
β-carotene and
cryptoxanthin (all
β-
cultivars)
cryptoxanthin
 Table 1. (continued-VI)

In vitro
Bioactive Effect on
Food Processing digestion Reference
compound bioaccessibility
method

Red tomatoes
↑ BAcs in paste and
(fresh, paste, TT: dried vs. fresh tomato
puree, juice, Kamiloglu et al.,
Lycopene Cooking Dialysis No differences in other
dried, chopped, 2014
(70-80ºC/5-70min) treatments vs. fresh
and chopped
tomato
and cooked)

Chromoplast:

No effect in β-carotene
TT: BAcs (orange carrot)
Red and
and lycopene (red
orange carrots, Chromoplast
carrot), but ↓ lycopene
red and atomic (65, 75, 85 and
β-carotene and BAcs (tomato) Palmero et al.,
red tomatoes 95ºC/10min) Solubility
lycopene Cell cluster: 2014
(chromoplast Cell clusters
and cell Low intensity TT ↓ β-
(95, 105, 115 and
clusters) carotene and lycopene
125ºC/25min)
BAcs, but high intensity
TT only ↑ β-carotene
BAcs

TT:
β- ↑ β-cryptoxanthin BAcs Aschoff et al.,
Orange juice Pasteurization Dialysis
cryptoxanthin 2015
(90ºC/54s)

TT:
Lutein,
Microwave
neolutein a+b,
Solubility and No effect on carotenoid Benlloch-Tinoco
β-carotene, Kiwifruit puree (1000 W, 340s)
dialysis BAcs et al., 2015
neoxanthin,
Pasteurization
violaxanthin)
(97ºC, 30s)

TT: ↓ BAcs (4%)

Boiling (5,10, In general, boiling gets

Pro-vitamin A 20min) Solubility and better BAcs than
α- and β- Ribeiro et al.,
biofortified Steaming (7,14, dialysis/Caco-2 steaming and boiling
carotene 2015
pumpkin 28min) cells with sugar added

Boiling + sucrose ↑ time of cooking ↑

addition (60%) BAcs

 Table 1. (continued-VII)

In vitro
Bioactive Effect on
Food Processing digestion Reference
compound bioaccessibility
method

In general, ↓ all

TT carotenoids BAcs, with

Violaxanthin any treatment, mainly

(90ºC/60s)
+neoxanthin, with TT, except ↑
Fruit juice, HPP
antheraxanthin, violaxanthin
milk-based
lutein, +neoxanthin in all Rodríguez-
(400
fruit beverages Solubility and
zeaxanthin, α- beverages and Roque et al.,
MPa/40ºC/5min)
and soy milk- dialysis
and β- antheraxanthin and 2016
based fruit HIPEF
cryptoxanthin, lutein in soymilk-fruit
beverages
α- and β- (35 kVcm-1, 4 s with HIPEF and HPP

carotene bipolar pulses, 200 and ↑ lutein in milk-fruit

Hz, 1800s) with HIPEF

Red and
yellow tomato- CEO-SP: No effect
based model
CEO-IP: ↓ carotenoid
system:
BAcs
carotenoid
CHRO-SP: No effect in
enriched oil
HPP: β-carotene BAcs and
(CEO) 5%,
β-carotene and Palmero et al.
100 MPa, 1 cycle Solubility slight ↑ in lycopene
chromoplast
lycopene 2016
BAcs
(CHRO) and
cell clusters CHRO-IP: ↑ carotenoid
(CC), BAcs
dispersed into
CC-SP and CC-IP:
soluble (SP)
and insoluble ↑ carotenoid BAcs

(IP) phases

Solubility and
Caco-2 cells/co- TT no effect on
Total TT:
culture cell carotenoids
carotenoids, Plum and Kaulmann et al.,
Boiling (7min) model: Caco-2: Carotenoid uptake was
xanthopylls, cabbage 2016
HT-29-MTX higher in co-culture vs.
carotenes Steaming (7min)
(mucus monoculture cell model
producing cells)
 Table 1. (continued-VIII)

In vitro
Bioactive Effect on
Food Processing digestion Reference
compound bioaccessibility
method

Mloukhiya β-Carotene: ↓ BAcs,

higher at TT 60/150 min
β-carotene and (Tunisian
TT
lycopene and dish): Dried Lycopene: not detected
Dhuique-Mayer
jute leaves + (95ºC/ 30, 60 and Solubility
lutein Lutein: ↑ BAcs, et al., 2016
tomato paste + 150min)
higher at TT 60/150 min
olive oil +
water
 Table 2. Effect of processing on measured bioaccessibility of minerals.

Bioactive In vitro digestion Effect on

Food Reference
compound Processing method bioaccessibility

↓ Ca with both treatments

in all foods
TT:
White beans No changes in Fe (except
Traditional ↓ in beans traditionally- Sebastiá et al.,
Chickpeas Dyalisis
cooking cooked) 2001
Lentils
Microwave ↓ Zn in lentils (both
treatments) and chickpeas
(traditional cooking)

TT:

Traditional ↑ Ca cell uptake only in

Ca
cooking Solubility/Caco-2 ready-to-eat Viadel et al.,
Fe Lentils
cells ↓ Fe and Zn cell uptake 2006a
Industrial
Zn
processing (ready- with both food processing

to-eat)

↑ Ca and Zn cell uptake

White beans TT:
Solubility/Caco-2 in white beans and lentils Viadel et al.,
Chickpeas (traditional
cells ↓ Ca, Fe and Zn cell 2006b
Lentils cooking)
uptake in chickpeas

Cereals: rice,
↓ Zn in cereals (except
finger millet,
wheat and sorghum) and
sorghum, wheat
pulses by pressure-
and maize
TT: cooking and microwave
Fe Hemalatha et al.,
Pulses:
(pressure-cooking Dialysis ↑ Fe in cereals and pulses
Zn chickpea, green 2007
and microwave) by pressure-cooking and
and black and
microwave (except
red gram,
sorghum, maize, chickpea
cowpea and
and French bean)
French bean

TT:
Milk-based
↑ Ca solubility in HPP vs.
(whole milk, (90ºC/30s)
Ca Solubility/Caco-2 TT samples
skimmed milk HPP: Cilla et al., 2011
P cells ↑ P solubility and cell
and soya milk)
(400 MPa/40ºC/ uptake in HPP vs. TT
fruit beverages
5min)
TT: Thermal treatment. HPP: High pressure processing
Solubility method: Determination of the soluble component fraction obtained by centrifugation to obtain the
bioaccessible fraction

Dialysis method: Dialysis of soluble components across a semi-permeable membrane to obtain the bioaccessible
fraction
 Table 2. (continued-I).

Bioactive In vitro digestion Effect on

Food Processing Reference
compound method bioaccessibility

↓ Ca soluble and
dialyzable

Fe = untreated (except ↑
soluble/4min and
HPP:
Apples Solubility and dialyzable/8min; ↓ Briones-Labarca
500 MPa/20ºC/ 2,
(Granny Smith) dialysis dialyzable/4min) et al., 2011a
4, 8 and 10min
Zn = untreated (except ↑
soluble/2min and
Ca dialyzable/8-10min; ↓
Fe dialyzable/2-4min)

Zn Ca ↑ solubility and ↓
dialyzabilitty
Algarrobo HPP:
Solubility and Fe ↑ solubility and Briones-Labarca
(Prosopis 500 MPa/20ºC/ 2,
dialysis dialyzability/10min et al., 2011b
chilensis) seeds 4, 8 and 10min
Zn ↑ solubility/10min and
↓ dialysis/2-4-8min

Enteral TT: ↓ Ca, Fe and Zn after Galán and Drago,

Dialysis
formulas Boiling boiling 2014

Fe: Pressure cooking ↑

TT:
BACs (and higher
Pressure cooking without soaking) vs.
(4 min, room TºC, cooking in a regular pan
without and with
Zn: In general, soaking
Fe previous soaking)
Pereira et al,
Cowpea beans Dialysis previous pressure cooking
and boiling in a
Zn 2016
did not affect BAcs, but it
regular pan (30
reduced BACs previous
and 10min,
cooking in a regular pan
without and with
previous soaking, ↑ Zn BAcs than Fe BAcs
respectively)
 Table 2. (continued-II).

In vitro
Bioactive Effect on
Food Processing digestion Reference
compound bioaccessibility
method

Cereals: wheat,
rice, finger
millet, maize,
sorghum and
pearl millet

Pulses:
chickpea,
TT ↓ BACs Se and
Se, cowpea, horse TT:
SeMet (except in
selenomethionine gram, green, Pressure-cooking microwaved black gram Khanam and
(SeMet) and black and red (10min) and Dialysis
and amaranth leaves) Platel, 2016
selenocysteine gram microwave
SeCys2 was destroyed
(SeCys2) (360W, 20min)
Green leafy
due to TT
vegetables:
fenugreek,
spinach,
amaranth, P.
graveolens and
Moringa
oleifera

46
 Table 3. Effect of processing on measured bioaccessibility of antioxidants (total
antioxidant capacity, ascorbic acid, tocopherols and polyphenols).

Bioactive In vitro digestion Effect on

Food Processing Reference
compound method bioaccessibility

HPP
TAC Apples Briones-Labarca
500 MPa/20ºC/ 2, Solubility ↑ TAC at 4-8-10min
(DPPH) (Granny Smith) et al., 2011a
4, 8 and 10min

Algarrobo HPP
TAC ↑ TAC with all HPP Briones-Labarca
(Prosopis 500 MPa/20ºC/ 2, Solubility
(DPPH) conditions 2-4-8-10min et al., 2011b
chilensis) seeds 4, 8 and 10min

TT
Milk-based
(whole milk, (90ºC/30s)
Ascorbic TT confers ↑ ascorbic
skimmed milk HPP Solubility
acid acid than HPP
and soya milk)
(400 MPa/40ºC/
fruit beverages
5min)
Cilla et al., 2012
TT
Milk-based HPP provides ↑ α-
(whole milk, (90ºC/30s) tocopherol in all samples
α-, γ and δ-
skimmed milk HPP Solubility and ↑ γ- and δ-
tocopherol
and soya milk) tocopherols in soya milk
(400 MPa/40ºC/
fruit beverages samples than TT
5min)

TT:

EGC Ambient

Green tea temperarute-

EC
↑ EGC, EGCG and ECG
with/without brewed
EGCG Solubility in h-GTE with/without Xie et al., 2013
milk (10% or (23ºC)(GTE)
milk vs. GTE
GCG 25%) Boiling-brewed
ECG (100ºC)(h-GTE)

DPPH: 2,2-diphenyl-1-picrylhydrazyl. EC: epicatechin. ECG: epicatechingallate. EGC: epigallocatechin. EGCG:

epigallocatechingallate. GCG: gallocatechingallate. GTE: green tea extract. h-GTE: green tea extract prepared at
boiling temperature. HPP: High pressure processing. TAC: total antioxidant capacity. TT: Thermal treatment.

Solubility method: Determination of the soluble component fraction obtained by centrifugation to obtain the
bioaccessible fraction

47
 Table 3. (continued-I).

Bioactive In vitro digestion Effect on

Food Processing Reference
compound method bioaccessibility

Total
polyphenols ↑ polyphenols and
Red tomatoes
and flavonoids Bacs and TAC
TT:
(fresh, paste,
flavonoids, in paste and dried vs.
puree, juice, Cooking Kamiloglu et al.,
Dialysis fresh tomato
TAC
dried, chopped, 2014
(70-80ºC/5-
(ABTS, No differences in other
and chopped
70min)
DPPH, treatments vs. fresh
and cooked)
FRAP, tomato
CUPRAC)

Finger millet: Pressure

cooking ↑ polyphenols
TT:
and flavonoids Bacs, but

Total
Roasting (150ºC, other TT ↓ polyphenols
Cereal:
5min), pressure Bacs and do not affect to
polyphenols Hithamani 
Finger millet cooking, boiling Dialysis
and flavonoids Srinivasan, 2014a
and pearl millet (5, 10, 15min) and
flavonoids Pearl millet: All TT ↑
microwave
polyphenols and
heating
flavonoids Bacs, except
roasting

Roasting: In general, ↓ or
no effect on Bacs

Pressure cooking: ↑ Bacs

in shorgum, wheat
TT:
Cereal: (polyphenols) and
Roasting (150ºC, chickpea (flavonoids)
Total
Wheat, shorgum
5min), pressure
polyphenols Boiling: ↑ Bacs of Hithamani 
Legumes: cooking, boiling Dialysis
and flavonoids in all of them Srinivasan, 2014b
(5, 10, 15min) and
flavonoids Green gram, and polyphenols in
microwave
chickpea shorgum
heating
Microwave heating: ↑
Bacs in wheat, shorgum
and green gram, but ↓
Bacs in chickpea
ABTS: 2,2-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid. DPPH: 2,2-diphenyl-1-picrylhydrazyl. FRAP: ferric
reducing antioxidant power. CUPRAC: cupric ion reducing antioxidant capacity

Dialysis method: Dialysis of soluble components across a semi-permeable membrane to obtain the bioaccessible
fraction

48
 Table 3. (continued-II).

Bioactive In vitro digestion Effect on

Food Processing Reference
compound method bioaccessibility

Total
↑ TAC (CUPRAC
polyphenols,
TT: method) only in jam
TAC Black carrot
Kamiloglu et al.,
(ABTS, jam and Boiling Dialysis ↑ phenolic acids Bacs in
2015
CUPRAC) marmalade (100ºC/30min) both samples
and phenolic
acids

Vitamin C ↑ with HPP in

TT
milk-juice and ↓ with TT
(90ºC/60s) in juice and soy milk-
juice. No effect with
HPP
Vitamin C, Fruit juice, HIPEF
total milk-based fruit (400MPa/40ºC/5min) Rodríguez-
Total polyphenols ↑ with
polyphenols beverages and Dialysis Roque et al.,
HIPEF HPP, HIPEF and TT in
and HAA soy milk-based 2015
(35 kVcm-1, 4s all beverages
(DPPH) fruit beverages
bipolar pulses, 200 HAA ↑ with HPP,
Hz, 1800s) HIPEF and TT in milk-
juice, but TT ↓ HAA in
juice and soy milk-juice

TT (90ºC/60 s)
LAA ↑ with HIPEF,

Fruit juice, HPP (400 HPP and TT in milk-

milk-based fruit MPa/40ºC/5min) juice, but ↓ soy milk- Rodríguez-

LAA Solubility and
beverages and juice. Roque et al.,
HIPEF
(DPPH) dialysis
soy milk-based In juice, HIPEF and HPP 2016
(35 kVcm-1, 4s
fruit beverages ↑ LAA, but TT had no
bipolar pulses, 200
effect
Hz, 1800s)

AJ:
Total
TT HPP ↓ TP BAcs, but TT
phenolics
Apple (AJ), no affect
(TP), (80ºC/30min,
grape (GJ) and
phenolic 90ºC/30s) Solubility GJ and OJ: He et al., 2016
orange juice
compounds
(OJ) HPP (250 TT (both) ↑ TP BAcs,
and TAC
MPa/10min) but HPP no affect
(ABTS)

HIPEF: High-intensity pulsed electric fields. HAA: Hydrophilic antioxidant activity LAA: Lipophilic antioxidant
activity

49
 Table 3. (continued-III).

Bioactive In vitro digestion Effect on

Food Processing Reference
compound method bioaccessibility

TT:
Total TP: ↑ in jam processing, ↓
Boiling (200ºC, in pickling
phenolics
40 min), drying
(TP), total
TF: ↑ in drying and jam
(80ºC, 8h),
flavonoids
pickling (200ºC, TAC: ↑ in jam
(TF) and Solubility and Guldiken et al.,
Red beetroot 20 min), jam (CUPRAC), ↓ in pickling
TAC dialysis 2016
processing
(ABTS, (FRAP), no determined
(200ºC, 15min)
DPPH, by DPPH and no
and puree
FRAP, differences between TT
processing
CUPRAC) by ABTS
(200ºC, 40min)

Solubility and
TT: Caco-2 cells/co-
Plum and culture cell model: TT ↓ polyphenols BAcs Kaulmann et al.,
Polyphenols Boiling (7min)
cabbage Caco-2: HT-29- 2016
Steaming (7min) MTX (mucus
producing cells)

Mloukhiya

(Tunisian dish): TT α-Tocopherol: ↓ BAcs, no

α- Dried jute Dhuique-Mayer
Solubility differences with TºC
(95ºC/ 30, 60 and
tocopherol leaves + tomato et al., 2016
150min)
paste + olive oil
+ water

50