Trends in Food Science & Technology 76 (2018) 28–37

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Trends in Food Science & Technology

journal homepage: www.elsevier.com/locate/tifs

An overview of the traditional and innovative approaches for pectin T

extraction from plant food wastes and by-products: Ultrasound-,
microwaves-, and enzyme-assisted extraction
Mirela Marića, Antonela Ninčević Grassinob, Zhenzhou Zhuc, Francisco J. Barbad,∗∗,
Mladen Brnčićb,∗, Suzana Rimac Brnčićb
a
Aviva Polyclinic, Zagreb, Croatia
b
Faculty of Food Technology and Biotechnology, University of Zagreb, Croatia
c
School of Food Science and Engineering, Wuhan Polytechnic University, 430023 Wuhan, China
d
Nutrition and Food Science Area, Preventive Medicine and Public Health, Food Sciences, Toxicology and Forensic Medicine Department, Faculty of Pharmacy, Universitat
de València, Avda. Vicent Andrés Estellés, s/n, 46100 Burjassot, València, Spain

A R T I C LE I N FO A B S T R A C T

Keywords: Background: A large amount of food wastes and by-products are produced from farm to plate. They represent
Pectin valuable sources for the production of high-added value compounds such as pectin. Pectin is the methylated
Food wastes ester of polygalacturonic acid and presents a wide range of applications in pharmaceutical and cosmetic products
By-products as well as in food industry such as gelling agent in fruit-based products, stabilizer in fruit and milk beverages and
Ultrasound
fruit filling for bakery and confectionary products, among others. Therefore, pectin recovery is of great im-
Microwaves
Enzyme-assisted extraction
portance.
Scope and Approach: The commercially available pectin is almost exclusively derived from citrus peels or apple
pomace, by-products from fruit juice manufacturing. But, nowadays the number of novel food waste and by-
products sources for pectin extraction are increasing. Moreover, the application of innovative approaches is
necessary due to the limitation of conventional processes. The present review will focus on the conventional and
innovative processing techniques (microwave extraction, enzymatic extraction, ultrasound-assisted extraction)
to extract pectin from different wastes and by-products.
Key Findings and Conclusions: The pectin extraction differs according to the matrix studied as well as tempera-
ture, pH, time, solvents, and solid to liquid ratio. The use of innovative extraction processes such as ultrasound,
microwave and enzymes can be a useful tool to increase pectin yield and quality, and reducing extraction time,
temperature, use of toxic solvents and strong acidic conditions for pectin recovery. Moreover, the combination of
solvent modelling and the use of particular extraction processes can enable the selective recovery of pectin.

1. Introduction
Food processing industry annually generates around 100 Mt of food
waste and by-products only in the EU, representing harvesting and
processing more than 35% of loss. Drink industry (26%), followed by
the dairy and ice cream industry (21.3%) and the production and
preservation of fruits and vegetables (14.8%) produce the largest
amounts of food wastes (Arshadi et al., 2016; Baiano, 2014). Significant
amounts of compounds with potential for valorization are lost during
overall processing and analysis like milling, drying, pasteurization,
extrusion, extraction, cooking and mixing (Brnčić, Ježek, Rimac Brnčić,
Bosiljkov, & Tripalo, 2008; Brnčić et al., 2009; Corrales, Toepfl, Butz,
Knorr, & Tauscher, 2008; Galanakis, 2012, 2013; Ježek et al., 2008).
The effective utilization of food waste and by-products has huge po-
tential for obtaining functional ingredients (Granato, Nunes, & Barba,
2017). For instance, valorization can be achieved by separating the
valuable compounds such as polysaccharides, polyphenols, essential
oils, pigments, proteins, flavour compounds, enzymes, dietary fibres
using different extraction methods (Barba, Zhu, Koubaa, Sant’Ana, &
Orlien, 2016; Koubaa et al., 2016; Parniakov, Barba, Grimi, Lebovka, &
Vorobiev, 2014; Roselló-Soto et al., 2016; Zhu et al., 2016).
Pectins are polysaccharides occurring in the cell walls located in the

∗
Corresponding author.
∗∗
Corresponding author.
E-mail addresses: francisco.barba@uv.es (F.J. Barba), mbrncic@pbf.hr (M. Brnčić).

https://doi.org/10.1016/j.tifs.2018.03.022
Received 10 January 2018; Received in revised form 12 March 2018; Accepted 28 March 2018
Available online 03 April 2018
0924-2244/ © 2018 Elsevier Ltd. All rights reserved.
M. Marić et al.
middle lamella, and primary and secondary cell wall of plant tissue. D-
galacturonic acid (D-GalA), an isomer of D-glucuronic acid (D-GlcA)
was found to be a basic constituent of all pectins. It is present in three
polymeric forms, i.e. homogalacturonan (HG), a linear polymer of α-1-4
linked galacturonic acid, rhamnogalacturonan I (RG-I), a repeating
disaccharide of galacturonic acid and rhamnose and rhamnoga-
lacturnon II (RG-II), a homogalacturonic backbone with numerous
complex side chains containing rhamnose and other neutral sugars
(Mohnen, 2008; Yapo, 2011). To date, pectins are thought to be com-
posed of at least of 17 kinds of monosaccharides, being D-GalA the
predominant one, followed by D-galactose or L-arabinose (Yapo, 2011).
HG is the most abundant pectic polysaccharide in plant cell walls. It is
composed of a linear chain of (1,4)-linked α-D-GalA units, in which
some of the C-6 carboxyl groups are methyl-esterified. In this context,
the degree of methoxylation is defined as the percentage of esterified
galacturonic acid units of the total number of galacturonic acid units in
the molecule.
Pectin structure determines in a great manner its physicochemical
properties and applications. The degree of esterification of pectin is
important to the functional properties in the plant cell wall and also
affects its application as gelling and thickening agent. According to
Hosseini, Khodaiyan, and Yarmand (2016a, 2016b), the degree of es-
terification (DE) can be defined as the percentage of carboxyl groups
esterified with methanol. On the other hand, Gnanasambandam and
Proctor (2000) established the degree of esterification as the percentage
of the total number of carboxyl groups esterified or methoxyl content of
total pectin. Moreover, Flutto (2003) determined that the degree of
esterification is the percentage of esterified galacturonic acid units of
the total number of galacturonic acid units in the molecule.
For instance, pectin is divided into two major groups depending on
the degree of esterification: high methoxyl pectin, with degree of es-
terification higher than 50% and low methoxyl pectin, with degree of
esterification lower than 50% (Thakur, Singh, & Handa, 1997) (Fig. 1).
These different forms of pectin have several applications. High meth-
oxyl pectin form gels when heated in acid solutions with a low pH
(2–3.5) and in the presence of high concentrations of sugar (55–75%).
Low methoxyl pectin can form a gel in a wide range of pH (2–6) with or
without a little amount of sugar in the presence of divalent ions such as
Fig. 1. Structures of a) high and b) low methoxyl pectin.
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Trends in Food Science & Technology 76 (2018) 28–37
calcium (Ca2+) (Fishman, Cooke, Chau, Coffin, & Hotchkiss, 2007).
High and low methoxyl pectins have different physicochemical
properties, and thus diverse applications. High methoxyl pectin can be
used as a gelling agent, stabilizer, emulsifier and thickener in the food
industry for the production of jams and jellies while low methoxyl
pectin can be used like a fat replacer in spreads, ice cream, fruit pre-
paration for yoghurt, heat reversible bakery glazing, emulsified meat
products or low calorie products such as diet carbonated beverages.
Over the last years, pectin has led to an increased interest as po-
tential emerging prebiotic with improved properties, such as their su-
perior ability for modulating microbiota and for reaching and causing
effects on the distal part of the colon (Gómez, Gullón, Yáñez, Schols, &
Alonso, 2016; Li, Xia, Nie, & Shan, 2016). The global hydrocolloids
market has shown that pectin is a fast growing functional ingredient in
many areas such as food, cosmetics, animal feed, edible packaging
films, personal care products and pharmaceutical industries.
The main sources for commercial pectin production are citrus peels
(Putnik et al., 2017), apple pomace (Kumar & Chauhan, 2010) and
sugar beet pulp (Chen, Fu, & Luo, 2015). Moreover, pectin from non-
conventional sources such as cocoa husks (Chan & Choo, 2013), mul-
berry branch bark (Liu, Jiang, & Yao, 2011), faba bean hulls (Korish,
2015), sisal waste (Santos, Vieira, Braz-Filho, & Branco, 2015), water-
melon rind (Prakash Maran, Sivakumar, Thirugnanasambandham, &
Sridhar, 2014), pomegranate peels (Pereira et al., 2016), pumpkin
(Ptichkina, Markina, & Rumyantseva, 2008) and banana peel (Oliveira,
Giordani et al., 2016) has been also evaluated. In the industry, com-
mercially available pectin is obtained chemically using acid extraction
but some innovative extraction approaches have been developed to
improve ectraction process and pectin quality. Some of the most re-
levant findings are reported below.
2. Extraction methods
Several methods have been applied to extract pectins from different
plant sources. The utilization of a suitable method for pectin extraction
is significant to maximize its extraction yield and improve the product
quality. Among the extraction techniques, there are conventional
techniques such as extraction with the mineral acids (Table 1, Fig. 2)
M. Marić et al. Trends in Food Science & Technology 76 (2018) 28–37

Table 1
Effect of conventional chemical extraction conditions on pectin yield from several plant food materials, wastes and by-products.
Plant material Treatment Treatment Extraction solvent Yield Reference
conditions (%)

°C min

Orange peel MA 82 50 0.5% HA, pH 1.7 2.95 Georgiev et al. (2012)

Lemon peel MA 82 50 0.5% HA, pH 1.7 3.15 Georgiev et al. (2012)
Peach pomace MA 60–80 10–80 NA, pH 1.2–2.53 2.6–18.0 Pagán, Ibarz, Llorca, Pagán, and Barbosa-Cánovas (2001)
Banana peels OA 70–90 120–240 Citric acid, pH 2.0–4.5 5.2–12.2 Oliveira, Giordani et al. (2016) and Oliveira, Rosa, et al. (2016)
Passion MA 70–98.7 30–90 HA, pH 1-3 7.48–14.80 Kulkarni & Vijayanand (2010)
fruit peel
Passion fruit peel MA 50 5–60 1.0 M NA, 4.8–6.2 Oliveira, Giordani et al. (2016) and Oliveira, Rosa, et al. (2016)
2.0 pH 2.0
Passion fruit peel MA 35–85 30–120 0.1 N Citric acid 3.68–7.71 Liew, Chin, Yusof, and Sowndhararajan (2016)
Pomegranate peels OA 70–90 40–150 Citric acid, pH 2-4 3.92–11.18 Pereira et al. (2016)
Carrot pomace OA 50–90 30–150 Citric acid, 5–15.2 Jafari et al. (2017)
pH 0.5–2.5
Apple pomace MA 85 180 Sulphuric acid, pH 2.0 14.5 Wikiera et al. (2015a, b)
Grapefrit peel MA 90 90 0.1 N HA 19.16 Bagherian, Zokaee Ashtiani, Fouladitajar, and Mohtashamy (2011)
Kiwifruit OA 50 60 1% Citric acid, pH 2.20 3.83 Yuliarti et al. (2015)
pomace
Pistachio gren hull OA 50–90 30–150 Citric acid, 7.31–19.02 Chaharbaghi, Khodaiyan, and Hosseini (2017)
pH 0.5–2.5
Husk tomato waste MA 100 15–25 0.1 N HA 9.6–19.8 Morales-Contreras et al. (2017)
Melon peel OA 35–95 40–200 Citric acid, pH 1-3 2.87–28.98 Raji, Khodaiyan, Rezaei, Kiani, and Hosseini (2017)
Potato pulp MA 90 60 HA, pH 2.04 9.72 Yang et al. (2018)
Potato pulp MA 90 60 Sulphuric acid, pH 2.04 8.38 Yang et al. (2018)
Potato pulp MA 90 60 NA, pH 2.04 9.83 Yang et al. (2018)
Potato pulp OA 90 60 Citric acid, pH 2.04 14.34 Yang et al. (2018)
Potato pulp OA 90 60 Acetic acid, pH 2.04 4.08 Yang et al. (2018)

MA: Extraction using mineral acids. OA: Extraction using organic acids. HA: Hydrochloric acid. NA: Nitric acid.

Fig. 2. An schematic representation of pectin extraction process assisted either by conventional and non-coventional processes. CE: Conventional extraction. NCE:
Non-conventional extraction. EAE: Enzyme-assisted extraction. MAE: Microwave-assisted extraction. UAE: Ultrasound-assisted extraction.

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M. Marić et al. Trends in Food Science & Technology 76 (2018) 28–37

Table 2
Effect of ultrasound extraction conditions on pectin yield from several plant food materials, wastes and by-products.
Plant material Frequency (kHz) Power Treatment conditions Extraction solvent Yield (%) Reference
(W)
Temperature °C Time min

Pomegranate peel 20 130 W 50–70 15–35 Distilled water, pH 1-2 4.23–24.18 Moorthy et al. (2015)
Grape pomace 37 35–75 20–60 Citric acid, 3.19–29.38 Minjares-Fuentes et al. (2014)
140 W pH 1-2
Tomato waste 37 60–80 15–90 Ammonium oxalate (16 g L -1)/oxalic 15.1–35.7 Grassino et al. (2016)
acid (4 g L−1)
Grapefruit peel 20 60–80 20–40 0.5 M Hydrochloric acid l, pH 1.5 23.10–27.51 Wang et al. (2015)
800 W
Grapefruit peel 20 30–80 10–60 0.5 M Hydrochloric acid, 10–18.11 Xu et al. (2014)
800 W pH 1.5
Grapefruit peel 24 50–70 4–30 0.1 N Hydrochloric acid, 13.51–17.92 Bagherian et al. (2011)
200 W pH 1.5
Passion fruit peel 20 45–85 2–20 1M nitric acid, pH 2.0 7.53–12.67 Oliveira, Giordani et al. (2016) and Oliveira,
750 W Rosa, et al. (2016)

Table 3
Effect of microwave extraction conditions on pectin yield from several plant food materials, wastes and by-products.
Plant material Power (W) Treatment conditions Solvent Yield (%) Reference

Time
Min

Pomelo peel 390–650 3–7 Hydrochloric acid, 0.05–2.93 Chen et al. (2016)
pH 1-3
Sour orange peel 300–700 1–3 Citric acid, 5.2–26.4 Hosseini et al. (2016a, 2016b)
pH 1.5–3
Pumpkin biomass 1200 2–10 35% Hydrochloric acid, 3.1–7.4 Košťálová, Aguedo, and Hromádková (2016)
pH 2.5
Sugar beet pulp 150–250 2–4 Sulphuric acid, 5–32.4 Li et al. (2012)
pH 1-2
Papaya peel 320–640 0.5–3 pH 1-3 2.49–24.69 Prakash Maran & Prakash (2015)
Orange peel 160–480 1–3 Sulphuric acid, pH 1-2 7.42–18.59 Prakash Maran, Sivakumar, Thirugnanasambandham, and Sridhar (2013)
Water melon rind 160–480 1–3 Hydrochloric acid, 13.09–24.18 Prakash Maran et al. (2014)
pH 1-2
Dragon fruit peel 150–550 10–30 distilled water 4.02–7.42 Thirugnanasambandham et al. (2014)
Banana peel 300–900 1.5–5 Hydrochloric acid, 1.09–2.17 Swamy and Muthukumarappan (2017)
pH 1-3
Passion fruit peel 356–450 3–9 Tartaric acid, 30 Seixas et al. (2014)
pH 2

Table 4
Effect of enzyme-assisted extraction on pectin yield and degrees of esterification from several plant food materials, wastes and by-products.
Plant material Enzyme Treatment conditions Yield (%) DE (%) Reference

°C h

Apple pomace Celluclast 1.5L 50 18 14.5 – Wikiera et al. (2015a, b)

Apple pomace Celluclast 1.5L 40 3 4.01–6.53 – Wikiera et al. (2015a, b)
Apple pomace Econase 40 3 3.63–4.22 – Wikiera et al. (2015a, b)
Apple pomace Viscoferm 40 3 4.67–7.00 – Wikiera et al. (2015a, b)
Kiwifruit pomace Celluclast 1.5L 25 0.5 4.48 – Yuliarti et al. (2015)
Passion fruit peel Celluclast 1.5L 35–85 0.5–2 2.59–9.17 52.6–86.9 Liew et al. (2016)
Lime peel Laminex C2K 50 4 22.5 82.2 Dominiak et al. (2014)
Lime peel Validase TRL 50 4 26.3 79.1 Dominiak et al. (2014)
Lime peel Multifect B 50 4 17.7 79.3 Dominiak et al. (2014)
Lime peel GC880 50 4 17.9 78.4 Dominiak et al. (2014)
Rapeseed cake Celluclast1.5L 50 4.5 5.41 45.77 Jeong et al. (2014)

and innovative techniques, such as ultrasound-, microwave-, and
enyme-assisted extraction with different mechanisms (Tables 2–4).
2.1. Conventional methods
Traditionally, pectin is extracted in water acidified with 0.05–2M
sulfuric, nitric, phosphoric, acetic or hydrochloric acid between 80 and
100 °C for 1 h with continuous stirring (Georgiev, Ognyanov,
Yanakieva, Kussovski, & Kratchanova, 2012). Conventional extraction
(solid-liquid extraction) depends on numerous factors such as tem-
perature, pH value, solvent properties, solid to solvent ratio, dry solids,
particle size, and diffusion rate. The maximum pectin yield can be
obtained at pH 2.0 using hydrochloric acid (Kulkarni & Vijayanand,
2010) (Table 1). However, conventional extraction using mineral acids
for the recovery of pectin has some important drawbacks such as losses
of some volatile compounds, increased costs for manufacturers and is-
sues from an environmental point of view as well as the degradation of
targeted valuable compounds found in plant materials. These

31
M. Marić et al.
shortcomings have lead to consideration of using some other solvents
and techniques. In this line, organic acids such as citric and acetic acid
have attracted considerable interest (Yang, Mu, & Ma, 2018). Although
strong mineral acids are cheaper and more effective than organic acids
to extract pectin, organic acids seem to be more interesting from an
environmental point of view. Organic acids have a lower hydrolyzing
capacity than mineral acids (because of their lower dissociation con-
stant), and are therefore expected to cause less depolymerization of
pectins. The fundamentals of conventional methods are different from
innovative extraction approaches since the extraction occurs due to
changes in the cell structure caused by electromagnetic or sound waves.
Pectin structure undergoes modification during extraction due to re-
action with extractants or other solutes. As a result, different extraction
methods can produce pectin with various structure features. Some of
the most relevant and interesting innovative approaches are shown
below.
2.2. Ultrasound assisted-extraction
Sound waves consist of mechanical vibrations, which can be applied
in treatments to the solid, liquid, or gas with frequencies higher than
20 kHz (Roselló-Soto et al., 2015; Zinoviadou et al., 2015). Ultrasound
represents the sound that is above the hearing of human being since the
sound that could be heard by humans goes from 16 Hz up to 16 kHz and
is mainly characterized by frequency (kHz range – MHz range) and
wavelength (Šic Žlabur et al., 2015). This technique is used for various
food processing purposes including extraction, drying, emulsification
and homogenization, as well as for microorganism and enzyme in-
activation, among others (Herceg et al., 2009; Misra et al., 2017).
For extraction purposes, the collapse of cavitation bubbles near cell
walls induced by ultrasound produces cell disruption, thus causing
stronger and enhanced solvent entrance into the cells and intensifica-
tion of the mass transfer (Barba, Brianceau, Turk, Boussetta, &
Vorobiev, 2015; Misra et al., 2017; Tiwari, 2015; Zhu, Wu, et al., 2017).
Besides the shock wave could facilitate swelling and hydration, and as
consequence the larger pores in the cell wall of the plant and structural
modification of plant tissue occurred (Zhu, Guan, et al., 2017). That is
how the diffusion of the inner cell material is promoted and mass
transfer increased. There are significant benefits of the ultrasound
versus conventional heating methods for the extraction such as low
energy consumption, shortened treatment time, less solvent usage, in-
creased safety of the operators, increased yield and in environmental
terms it is non-thermal technology (Chemat et al., 2017; Roselló-Soto
et al., 2015). Due to their ability to enhance extraction processes, ul-
trasound has been used to extract pectin from different plant food
materials, wastes and by-products. Some of the most important findings
are reported below and in Table 2.
During pomegranate (Punica granatum) processing, high amounts of
by-products are produced. These by-products mainly consist of peel
(78%) and seeds (22%). Besides being an important source of anti-
oxidants, pomegranate by-products have been also a core of the re-
search for the extraction of pectins (Moorthy, Maran, Surya,
Naganyashree, & Shivamathi, 2015). In order to extract pectins, ultra-
sound-assisted extraction (UAE) was conducted using an ultrasound
device of 20 kHz frequency with an immersed sonotrode and maximum
power of 130 W. The used probe was a flat tip with a 2-cm diameter.
After the extraction was carried out, the experimental results were
further analysed over the process variables (solid-liquid ratio, pH, ex-
traction time and temperature). Authors obtained an excellent corre-
lation between the predicted values and the experimental ones, thus
concluding that the highest pectin extraction yield was obtained after
using a solid-liquid ratio of 1:18 g/ml with processing parameters of
extraction time of 29 min and extraction temperature of 62 °C, respec-
tively.
In another study, UAE of pectin from grape pomace was the matter
of the research conducted by Minjares-Fuentes et al. (2014). They used
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Trends in Food Science & Technology 76 (2018) 28–37
RSM as a tool to optimize the extraction process. In addition, the Box-
Behnken Design (BBD) was precisely used to establish the appropriate
temperature, duration of extraction and pH to obtain a desirable yield
of extracted pectins with high average molecular weight and degree of
esterification (DE). In this research, citric acid (CA) was used as ex-
tracting agent to replace the conventional used mineral acids, and the
authors obtained a higher extraction of homogalacturonan-like poly-
saccharides compared to the conventional methodology. The results
found using the model were confirmed after being compared with the
experimental ones. Moreover, the predicted RSM values were coherent
with the experimental data. The RSM model predicted pectin extraction
yield, which was ≈3.2–29.4% with average molecular weight of the
110–205 kDa and DE of 20.1–62.1%. Furthermore, when UAE was
compared to the conventional extraction methods at equivalent tem-
perature (75 °C), time (60 min) and pH (2.0), the authors obtained an
increased molecular weight average and yield of extracted pectin (20%
more) when UAE was used.
On the other hand, the effects of conventional heating extraction
(CHE) and ultrasound-assisted heating extraction (UAHE) on pectin
yield and swelling behaviour of the grapefruit peel were evaluated (Xu
et al., 2014). The most promising results were found when the authors
used UAHE (25.9 min), obtaining up to 26.74% pectin yield, while the
maximum pectin yield determined using CHE as processing technology
was 23.44% after 72.26 min. When recalculated, the difference in
pectin yield was 12.35% higher for the UAHE compared with CHE.
Besides the temperature was 20 °C lower for the UAHE during the ex-
traction procedure and the treatment time was 39.53% shortened,
which gives opportunity for the UAHE approach as industrially poten-
tial for the future. At the end, the authors concluded how the main
mechanism of ultrasonic cavitation significantly damaged vegetable
tissue, which resolved as both improved contact of the solvent with the
inner cell material and better diffusion of the material after the contact
with the solvent.
Another similar research with the same by-product was conducted
by Wang et al. (2015). The UAHE and CHE were used to extract pectin
and the results were compared. The experiment was performed under
constant ultrasonic frequency of 20 kHz and cycle of 50%. Authors es-
tablished a model for extraction and gained excellent correlation with
the predicted grapefruit peel pectin yield and the obtained one
(27.46%/27.34%). They concluded how UAHE produced 16.34%
higher amount of the extracted pectin, shortened time by 37.78% and
decreased temperature up to 13.3 °C compared to CHE.
Tomato wastes and by-products, which mainly consist of seeds, skin
and pulp, have also attracted the interest of both food industry and
researchers due to their high content in valuable compounds, and
particularly pectin. For instance, Grassino et al. (2016) used two dif-
ferent methods to extract pectin out of tomato waste. Waste undergoes
conventional extraction (CE) and UAE. UAE was performed in dual
frequency using an ultrasonic bath Elma – Elmasonic P 300H (37/
80 kHz). In order to obtain stronger agitation, the lower frequency
(37 kHz) was applied for the full research. The CE was conducted in two
steps. The first step was performed under reflux and last for 24 h with
second step, where fresh volume of solvent was used. The UAE was
conducted during 15, 30, 45, 60 and 90 min. The obtained results
showed an increased pectin yield after CE at 60 °C compared to UAE
treatment but a shorter extraction time (15 min) was needed for UAE.
Moreover, UAE produced pectin of better quality (anhydrouronic acid,
methoxy and calcium pectate contents and degree of esterification). The
obtained extracts were analysed with the NMR and FTIR spectroscopy
and in all investigated samples the existence of predominantly ester-
ified pectin was observed. After the comparison of results was con-
ducted, it was noticed pectin yield was more or less the same for both
methods at 80 °C but with significant shortening of the treatment time
(CE/36 h and UAE/15 min). UAE extraction of pectin was found useful
and gives the opportunity to use an eco-friendly approach at reduced
temperature (compared to CE) and with significant savings of energy.
M. Marić et al.
Other plant material with huge interest is that obtained from sugar
industry. Sugar beet pulp (SBP) is generated in huge amounts as by-
product during sugar production. Subcritical water in combination with
ultrasound was used to extract pectin out of SBP (Chen et al., 2015). In
this work, the authors used a RSM design to optimize process para-
meters for obtaining an appropriate yield of pectin. UAE was applied
with frequency of 25 kHz and 10 min. Results showed the maximum
pectin (24.63%) under the processing conditions of extraction time
(30.49 min), extraction pressure (10.70 MPa), liquid/solid ratio (44.03)
and extraction temperature (120.72 °C). Moreover, the composition of
the pectin was studied as well, obtaining 59.12% of galacturonic acid
(GalA) and 21.66% of arabinose (Ara). Furthermore, the flow beha-
viours were evaluated with influence of pectin on both pasting and
thermal properties of maize starch. It was concluded that pasting
temperature was increased with the addition of pectin and all other
pasting properties were significantly decreased. Moreover, the pectin
addition significantly increased the gelatinisation temperature as well
as enthalpy.
2.3. Microwave-assisted extraction
Microwaves (MW), as processing tool, has played an important role
in the food science and technology over the last two decades. It can be
easily found in laboratories and industry as technology for the i) drying
of foodstuffs, ii) microwave-assisted extraction (MAE), iii) enzyme in-
hibition and inactivation, microorganism inactivation, and especially as
good choice for the processing of the various wastes and by-products
generated in food industry (Bouras et al., 2015; Deng et al., 2015; Sahin
et al., 2017). From the fundamental science, MW belong to non-ionizing
radiation with frequencies of starting 300 MHz up to 300 GHz (Barba
et al., 2016; Koubaa et al., 2015; Roselló-Soto et al., 2016). While en-
tering into food matrix, MW generates heat after getting in contact with
the cellular polar compounds. The generated heat reacts with the mo-
lecules causing ionic conduction and dipoles rotation (Wang, Ding, &
Ren, 2016). Ionic conduction is based on electrophoretic transfer of ions
and electrons initiated by the MW energy, which generates electric field
responsible for the particle movements, while alternate displacement of
polar molecules refers to dipoles rotation. Accordingly, the rotation
displacement of the polar molecules starts in the moment when the
molecules are in effort to align with existing electric field. This two
coupled mechanisms create synergy and promotes the release of energy
with the consequence of prompt heating. This is an internal heating,
thus promoting both rapid material damage and the increased diffusion
of the targeted compounds. During the propagation of the MW, the
molecules in contact absorb the energy, being the amount of energy
delivered higher or lower according to the dielectric constant.
Throughout the MAE as dielectric constant is higher, the molecule will
absorb more energy resulting in increased heating of the solvent and
giving the MAE at higher temperatures. On the other hand, while the
targeted extracted compounds do not withhold high temperatures the
solvents with low polarity are applicable. Under such MAE the heat
would disrupt the cell walls with the expansion mechanism and the
diffusion of desired valuable compounds out of the cellular material
would be directed towards the cooler solvent. The usage of MAE is
applicable for various plant materials in order to get valuable bioactive
compounds. MAE is an innovative extraction technology and combines
MW with traditional extraction procedures involving solvents as well
(Carbonell-Capella et al., 2017; Périno, Pierson, Ruiz, Cravotto, &
Chemat, 2016). In general, the conventional extraction technologies
require huge amounts of solvents with prolonged treatment time while
MAE is in favour on account of the shortened time of processing, sig-
nificant increase in yield, lower energy consumption and the decreased
usage of solvents. Therefore, being MAE an environmental friendly
solution in processing (Chemat et al., 2017).
The extraction of pectin from the food waste and by-products as-
sisted by MAE has been the matter of interest of several research groups
33
Trends in Food Science & Technology 76 (2018) 28–37
(Table 3). As already mentioned, MAE possess many advantages over
the traditional technologies, and with careful optimization of the pro-
cess it could be even more acceptable and feasible.
Citrus is a major processed crop that results in large quantities of
pectins (Putnik et al., 2017). Among citrus fruits, pomelo (Citrus
grandis) is mostly cultivated in Southeast Asia. Besides being rich in
various compounds like carotenoids, vitamin E, phenolic compounds,
lignin, fiber and essential oils, the pomelo fruit is also abundant in
pectins. Together with mentioned bioactive compounds in the fruit the
by-products generated out of pomelo fruit contain high amounts of
terpenoids and pectins as well (Chen, Hu, Yao, & Liang, 2016). The
pectin in pomelo peels is mainly found in the cell walls and has some
health-related properties (eg. ability to decrease the level of choles-
terol) (Voragen, Coenen, Verhoef, & Schols, 2009). In addition, pectin
with the peel origin has been classified as low-methoxyl one, being thus
spread for the production of the low-sugar products. In the work
mentioned above, the pectin extraction undergoes microwave extrac-
tion but also classical conventional extraction was conducted for the
comparison of results. Both microwave and conventional procedures
were carried out as two-step processes. The first step was used to extract
essential oils by solvent-free microwave extraction (SFME) or conven-
tional extraction hydrodistillation (HD) and the second step was ap-
plied to get pectin using now oil-free pomelo peels. Hot-solvent mi-
crowave extraction (HSME) was carried out for pectin extraction and
compared with conventional acidic solution extraction (CASE). The first
benefit observed was a significant shortening of treatment time to ob-
tain pectins using HSME compared to CASE (HSME/5.6 min vs CASE/
90 min). Moreover, under the processing conditions (520 W, pH = 2.0
value and 5.6 min) the yield of extracted pectin was 3.29% higher using
HSME than that obtained with CASE. Optical and scanning electron
microscopy studies were conducted to evaluate the morphological
changes of peels after using MW, obtaining positive results.
In another study, the effects of MAE on the extraction of pectin from
lime were evaluated. It should be taken into account lime is one of the
prevalent citrus fruit in India (Thirugnanasambandham & Sivakumar,
2015), thus having an important amount of wastes and by-products
generated in the process from farm to plate. The fruit is relatively small
in diameter (2–5 cm) with light green colour and contains sour pulp. As
abundant source of vitamin C, lime is used in food industry to accent
the flavors. Although the lime possess lower amount of the vitamin C in
comparison with lemmon, it is still widely used. By making lime pro-
cessed the waste is emerged in form of bagasse, which remains in
thousands of tones since only in India more the 500 industrial facilities
are developing products out of lime. The bagasse contains pectin, which
could be recovered by various extraction methods. For instance, MAE of
pectin from lime bagasse was evaluated (Thirugnanasambandham &
Sivakumar, 2015). MAE was conducted in a microwave oven and the
process optimization was carried out in terms of treatment time, MW
power, pH and temperature. Prior to experimental part the RSM was
employed to predict optimal conditions with the objective of pectin
yield estimation. As suggested, the process was conducted to obtain
pectin using MAE, thus for the model developing the yield of the pectin
was used as response value while the MW power, temperature, pH and
the sample mass were taken into account as independent variables. The
statistical analysis was carried out by ANOVA in Design-Expert pro-
gramme. The authors observed that MW power was a crucial factor for
the extraction of pectin from the lime bagasse, and therefore, the MW
power levels were used in the range from 100 to 500 W. This was
proven by the fact the pectin yield increased as MW power was in-
creased. Namely, when the power level was increased the solubility of
the sample was enhanced with the consequence of better extraction
productivity. The second important parameter to take into account
during MAE was the extraction time, which was studied under the
duration of 200–800 s. It was concluded that pectin yield was enhanced
in huge percentages when the treatment time was increased up to 600 s.
At the end, the authors concluded that even the pH and mass of the
M. Marić et al.
sample played a significant role in MAE extraction of lime bagasse
giving the optimal MAE conditions under the following processing
conditions (400 W, 500 s, pH = 1 and mass of the sample of 6 g). With
these optimal parameters the yield of the extracted pectin was 7.8 g/
100 g. This approach showed promising results in both terms of getting
valuable compound (pectin) and decreasing of level of the wasted
material out of lime processing.
Sour orange (Citrus aurantium) is a citrus fruit with a bitter taste
compared to other fruits from citrus family. This fact has a significant
influence on consumer's acceptance, thus being sour orange less at-
tractive for consumption. Therefore, it is more used as medicinal or
dietary supplement. After being processed in juice factories as by-pro-
duct, sour orange peel (SOP) occurs. It can be used for animal's feeding
but it can be additionally processed since it contains significant pectin
amounts. In the study of Hosseini et al. (2016a, 2016b), SOP undergo
MAE to obtain pectin. Authors used BBD to predict optimal conditions
of the MAE, thus decreasing large number of experiments. The influ-
ence of treatment time (seconds), microwave power (W) and pH as
independent variables was established in order to foresee response in
form of pectin yield (%). The achieved results correlated in a very good
manner with the predicted model. The optimal conditions for the ex-
traction of pectin from SOP were as follows: microwave power of
700 W, 180 s of treatment time, and pH = 1.5. Such conditions resulted
in 29.1% of extracted pectin out of SOP. Short time of MAE procedure
coupled with significant yield of pectin showed the potential of MAE to
be used as an eco-friendly solution for the arisen environmental pro-
blem represented in huge amounts of generated waste and by-products
from food industry and in this case SOP.
On the other hand, one of the interesting tropical fruit containing
high amounts of water-soluble-fiber and vitamin C is red pitaya
(Hylocereus polyrhizus), better known as dragon fruit. This wide spread
fruit in India comes from the cactus family of plants (Cactaceae). After
the processing of dragon fruit in juice industry huge amounts of dragon
fruit peels (DFP) remain. This by-product is good source of pectin. The
pure utilization of DFP as animal feed is insufficient in terms of en-
vironmental protection. Therefore, the DFP has been matter of the re-
search conducted by Thirugnanasambandham, Sivakumar, and Prakash
Maran (2014). MAE was applied to obtain pectin from DFP and RSM in
combination with the BBD employed for the optimization of MAE were
used. The DFP was purchased from the local juice processing factory.
Based on RSM, the authors selected MW power (W), temperature (°C),
duration of MAE (seconds) and solid-liquid ratio (g/mL) as independent
variables with only one response and it was the yield of extracted pectin
(%). The experimental results correlated well with predicted model and
the results showed the best yield of extracted pectin of 7.5% under the
following parameters: 400 W of MW power, 45 °C, 120 s and 24 g/mL of
solid-liquid ratio. It was shown and concluded that MAE was a suitable
technique for pectin extraction out of the DFP from both technological
and environmental point of view with promising industrial application.
Cocoa husks arise as by-product in chocolate manufacturing. Huge
amounts of cocoa husks remain after making the final product, thus
representing a significant environmental problem. Therefore, although
the pectin amount/yield in cocoa husks is lower per dry weight com-
pared to that obtained for citrus fruit peels, these by-products are still
very utilizable as they are also an important source of high-added value
compounds (eg. bioactive compounds). Furthermore, making cocoa
husks useful, environmentally friendly production can be achieved by
decreasing the overall amount of generated by-products. This by-pro-
duct has been processed by hot acid extraction (HAE) and using mi-
crowaves (MW) as pre-treatment (Mollea, Chiampo, & Conti, 2008). As
mentioned previously, HAE was used as a recovery tool for pectin ex-
traction out of cocoa husks. Mollea et al. (2008) obtained the best re-
sults under pH 2.5 and extraction duration of 60 min. Moreover, they
also designed and conducted the experimental work using MW as pre-
treatment. The results showed a significant improvement in pectin yield
(3.6%) after using MW compared to the samples without MW
34
Trends in Food Science & Technology 76 (2018) 28–37
pretreatment. Although it is not such benefit for the laboratory re-
search, the progress was obvious in terms of industrial processing
considering the lower energy consumption and the decrease in the
overall amount of waste.
The sugar beet pulp (SBPP) is also a very important by-product,
which in comparison with various citrus peels contain pectin with
higher degree of acetylation, larger neutral content of sugar, lower
molecular weight and feruloyl groups as well (Li, Jia, Wei, & Liu, 2012).
This by-product can be used in beverage industry for the purposes of
oil-in-water emulsions stabilization. Furthermore, if extracted the
pectin from SBPP, it could be useful in pharmaceutical industry in order
to develop drugs, flocculants and biosorbents. In their work, Li et al.
(2012) used MAE for pectin extraction from the SBPP. The SBPP was
purchased from an industrial supplier and exposed to MAE. The pro-
cedure was optimized by the RSM with BBD based on the inputs ob-
tained by previous experiences from the research group. The in-
dependent variables were as follows: MW power (150–250 W),
treatment time (2–4 min), pH of sulfuric acid (1–2) and solid-liquid
ratio (g/mL), while the response was monitored based on pectin yield
(%) and the average molecular weight (103) obtained by viscometry.
The obtained results indicated that the response of the yield and the
average molecular weight were of 5.2–32.4% for pectin yield and
7.8 × 103 to 315.3 × 103 for average molecular weight, respectively. In
addition, the authors concluded that the optimal pectin yield was ob-
tained under the conditions of 152.63 W (MW power), 3.53 min (MAE
extraction time), 1.57 pH of sulfuric acid and 18.92 solid-liquid ratio,
while the pH of sulfuric acid had the most significant influence on the
average molecular weight of SBPP.
The papaya (Carcia papaya L.) is a delicious fruit with origins of
South Mexico and Costa Rica. It is well known that papaya possess high
nutritive value and interesting bioactive composition (Parniakov et al.,
2015). Besides having nutritive value papaya fruit is widely used in
medicine for the purposes of preventing strokes, heart attacks and
diabetic heart diseases due to its high antioxidant and biological ac-
tivities. On the other hand, since the papaya production rate exceed 6
million tonnes per year in the world, enormous amounts of waste and
by-products are generated from farm to plate, including among them
papaya peel (WCCP) and seeds (Maran & Prakash, 2015; Parniakov
et al., 2014). The peels of papaya fruit are a rich source of various
bioactive compounds and can be used as untreated in households for
skin care protection. Yet the industrial recovery of the WCCP is required
because of huge environmental problems in disposal of WCCP coupled
with the benefits of the extracts which could be isolated. One of the
valuable compounds in papaya peels is pectin, which has the potential
to be used in pharmaceutical industry. On the account of everything
mentioned, the research of MAE of WCCP was carried out (Maran &
Prakash, 2015). The by-product in form of WCCP was taken from the
local industrial fruit juices company in India. Prior to the experimental
part was put into effect, the four factor experimental design with mi-
crowave power (W), pH, extraction time (seconds) and solid-liquid ratio
(g/mL) as independent variables and yield of extracted pectin out of
WCCP as response value was introduced. For this study, 29 experiments
were performed using MAE and to analyse results the analysis of var-
iance (ANOVA) was conducted. Full experiments were carried out with
experimental parameters of MW power (320–640 W), pH (1–3), ex-
traction time (20–180 s) and solid-liquid ratio (5–25 g/mL). The opti-
mized MAE extraction resulted in various yields between the lowest
(2.49%) up to the highest (26.69%). The best results considering the
above mentioned variables were achieved under the MW power of
512 W, pH = 1.8, 140 s and solid-liquid ratio of 1:15 g/mL, being the
obtained pectin yield from WCCP ≈25.41%.
2.4. Enzyme-assisted extraction (EAE)
The enzymes are used to improve extraction process by hydrolyzing
matrix of the plant cell wall. Enzymatic reactions disintegrate the cell
M. Marić et al.
wall and increase cell permeability. Enzyme aided extraction (EAE)
depends on reaction time, type and concentration of enzyme, tem-
perature, pH value and particle size of plant material (Poojary, Orlien,
Passamonti, & Olsen, 2017; Roselló-Soto et al., 2016) (Table 4). EAE
presents important benefits to extract pectin from plant food materials,
wastes and by-products. This technique can increase pectin extraction
yield compared to traditional processes and can be used under low
process temperatures, reducing energy consumption. EAE can be also
combined with conventional chemical solvent extraction, thus re-
quiring lower amounts of solvent and reducing the extraction time in
full correspondence with green extraction approaches. In addition, the
mild conditions used during EAE eliminate equipment corrosion, which
is frequently observed when high temperatures are used for extraction
purposes (Roselló-Soto et al., 2016; Saha et al., 2017). Moreover, EAE
does not require any additional step for waste neutralization and the
obtained pectin is of increased quality due to the high efficiency and
specifity of enzymes. However, one of the main drawbacks of EAE is the
cost of enzymes, which are relatively expensive for high amounts of raw
material. In addition, available enzymes cannot completely hydrolyze
plant cell walls, thus difficulting pectin extraction and it is difficult to
scale-up EAE process as several factors can influence enzymes response
(eg. dissolved oxygene, temperature, nutrient availability, among
others) (Saha et al., 2017).
The most commonly used enzymes during pectin extraction process
are cellulase, protease, hemicellulase, alcalase, xylase, poly-
galacturonase, α-amylase, neutrase, b-glucosidase, en-
dopolygalacturonase and pectinesterase (Jeong et al., 2014; Wikiera,
Mika, Starzyńska-Janiszewska, & Stodolak, 2015b; Yuliarti, Goh, Matia-
Merino, Mawson, & Brennan, 2015).
For instance, Yuliarti et al. (2015) compared the different extraction
methods of pectin from gold kiwifruit pomace and evaluated which was
the most advantageous in terms of pectin yield, protein, ash, non-starch
polysaccharide, galacturonic acid, neutral sugar composition, molar
mass, viscosity and degree of branching. The extraction methods in-
cluded acid extraction with 1% citric acid solution of pH 2.2 at 50 °C for
60 min, water extraction at 25 °C for 30 min and enzymatic extraction
at 25 °C for 30 min with Celluclast 1.5L. Celluclast 1.5L contains cel-
lulases, polygalacturonase, pectin lyase and rhamnogalacturonan lyase.
It breaks down the cellulosic materials of plant cell walls and converts
them to glucose, cellobiose and longer glucose polymers, thus being a
useful tool to extract pectin from gold kiwifruit. The optimum condi-
tions for Celluclast 1.5L activity are in the range of pH 4.5–6.0 and
temperatures ≈50–60 °C. The highest kiwifruit pomace pectin yield
was obtained by enzymatic extraction (4.5%). Possible explanations
include hydrolysis of cellulose and release of pectin trapped within the
cellulose matrix.
Wikiera et al. (2015b) compared the effectivenes of enzymatic ex-
traction of pectin using commercially available enzymatic preparations
of Celluclast 1.5L at a dose of 25, 50 and 75 μl per 1 g of apple pomace
for 18 h at 50 °C, pH 4.5 and acid extraction with sulphuric acid for
3 h at 85 °C. The obtained results showed that the lowest dose of Cel-
luclast 1.5L was more efficient (10.8%) than acid extraction (8.2%) for
pectin extraction. Wikiera, Mika, and Grabacka (2015a) also reported
an optimization of pectin extraction from apple pomace using three
different commercially available enzymatic preparations (Celluclast,
Econase and Viscoferm). Various pH values were tested (4.0, 4.5, 5.0
and 5.5) for finding the optimal exraction yield. Pectin yield depends on
the initial pH of the reaction mixture, and all the tested enzymatic
preparations were more efficient at pH 4.5. Moreover, several tem-
peratures were tested for optimizing pectin extraction, obtaining a
significant dependence between the optimal extraction temperature
and the enzyme preparation type (50 °C for Celluclast, 40–50 °C for
Viscoferm and 50–60 °C for Econase). Similarly to Celluclast, Viscoferm
and Econase are used to promote the degradation of plant cell wall due
to their high cellulolytic or xylanolytic activity, thus facilitating pectin
extraction. In addition, the enzymatic extraction was compared to the
35
Trends in Food Science & Technology 76 (2018) 28–37
conventional acid based extraction. The extraction yield achieved using
enzymatic preparations was higher (Celluclast 18.95% and Viscoferm
17.86%) than that obtained after acidic extraction (14.52%).
3. Conclusions
The application of UAE can lead to increased pectin yields with
decreased extraction time and temperatures. Moreover, the combina-
tion of ultrasound with heating (UAHE) could be also a useful tool to
reduce pectin extraction time from grapefruit peel and improving its
yield compared to conventional heating extraction (CHE). UAHE can
decrease the optimal temperature used in conventional heating ex-
traction to achieve the maximum pectin yield and can preserve better
the colour and microstructure of pectin compared to CHE. In addition,
hot-solvent microwave extraction was demonstrated to be superior in
both decreased extraction time and increased pectin yield compared to
conventional acidic solution extraction. Similarly, the use of enzyme-
assisted extraction improved pectin yield compared to control samples
(without enzymes). This fact can be attributed to the ability of enzymes
to induce cellulose hydrolysis, and the subsequent release of pectin
trapped within the cellulose matrix. At this stage of development, there
is a need to optimize and standardize the specific ultrasound-, micro-
wave and enzyme-assisted processing conditions according to the tar-
geted plant materials, food wastes and by-products in order to extract
the maximum pectin yield with the highest quality. Moreover, it is also
necessary to optimize non-specific extraction parameters (extraction
time, temperature, pH, origin of extracted material, material to solvent
ratio and number of extractions) because they will limit the amount and
quality of the extracted pectin, which will modify its physicochemical,
functional and technological properties, thus having a significant im-
pact in pectin potential applications.
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