Food Research 7 (1) : 211 - 223 (February 2023)

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Physicochemical characteristics, rheology, and emulsifying properties of
ultrasound-extracted pectin from 'saba' banana peel
1,2,*
Rivadeneira, J.P., 1Castillo-Israel, K.A.T. and 2Wu, T.
1
Institute of Food Science and Technology, College of Agriculture and Food Science, University of the
Philippines Los Baños, Laguna, Philippines 4031
2
Department of Food Science, The University of Tennessee, Knoxville, Tennessee, United States of America
37996
Article history: Abstract
Received: 1 October 2021
Received in revised form: 16
November 2021
Banana peel is an abundant agricultural waste in the Philippines. Valorization of the waste
Accepted: 28 February 2022 was done by extracting pectin from the 'saba' banana peel using ultrasound technology.
Available Online: 13 The ultrasound-extracted pectin (UEP) was characterized physicochemically. UEP was
February 2023 also compared with commercial low-methoxy pectin (LMP) in terms of rheology and
Keywords:
emulsification ability. Results showed that UEP has higher protein and ash content, and
Banana peel, lower methoxyl, total pectic content, and interfacial tension. During the emulsion
Ultrasound, preparation, increasing the concentration of both UEP and LMP resulted in an increased
Pectin, production of stable emulsion droplets. Also, UEP-stabilized emulsion had improved
Emulsifier
stability when higher oil fraction and ionic strength were used. In terms of pH, higher
DOI: volumes of stable emulsion were produced by UEP and LMP at conditions close to
https://doi.org/10.26656/fr.2017.7(1).770
pectin's isoelectric pH. Lastly, UEP and LMP produced emulsions that were stable in all
stages of in vitro digestion.

1. Introduction to medium-scale banana industry (Padam et al., 2014).

Banana is one of the major crops in the Philippines. There is increasing scientific evidence showing banana
In 2020, the Philippines produced around 9.05 million peels' physical properties, chemical compositions, and
metric tons of bananas. Of this volume, about 2.5 million nutritive potentials can be utilized to produce value-
metric tons is of the 'saba' variety (Figure 1). According added products for special food applications and
to the Philippine Statistics Authority (PSA) (PSA, nutraceuticals (Pathak et al., 2016). Analysis of the
2020a; PSA, 2020b; PSA, 2020c; PSA, 2020d), 25% or chemical composition of 'saba' banana peels has shown
more than 600,000 metric tons ‘saba’ is utilized for food that a relatively high amount of pectic content is present
processing. Of this, around 66% is composed of peel in the peels, which can be extracted as pectin
waste (Guerrero et al., 2016); hence, it is safe to say that (Rivadeneira et al., 2020). Between the unripe and ripe
the country produced more than 400,000 metric tons of banana peels, the peels generated by the earlier are 39%
waste in 2020. These wastes are often thrown away and cruder (Castillo-Israel et al., 2015).
end up in dumpsites. The accumulation of waste banana
The market for pectin on a global scale is estimated
peel could pose environmental problems as it generates
to account for a value of USD 1.0 billion in 2019. It is
methane and causes pollution.
expected to reach USD 1.9 billion by 2025 due to the
increasing demand for natural and organic products
(Markets and Markets, 2019). In the Philippines, data
from the Department of Trade and Industry showed that
about 95,000 kilograms were imported in 2011, valued at
around $52.4 million (Philippine News Agency, 2015).
Figure 1. 'Saba' banana Currently, the market price of commercial pectin
Waste banana peels are resources with low depends on the brand, but the lowest price is PhP 5000.
commercial value. However, it can be converted to It is mostly (85%) produced from citrus peels (56% from
highly commercial outputs that can improve the overall lemons, 30% from limes, and 13% from oranges), and
economics of processing units, particularly for the small apple pomace (14%), with a minor fraction being
*Corresponding author. eISSN: 2550-2166 / © 2023 The Authors.
Email: [email protected] Published by Rynnye Lyan Resources
 212 Rivadeneira et al. / Food Research 7 (1) (2023) 211 - 223

obtained from sugar beet. The world's largest producers Spencer Umfreville Pickering, who discovered the said
of dried citrus peel are in Argentina and Mexico, but phenomenon (Bon, 2014).
producers are also located in Peru, Spain, and Bolivia. In
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South America and Sicily, pectin is often extracted It was reported that pectin extracted from citrus
directly from the wet citrus peel after juicing. Brazil and reduced the interfacial tension of oil droplets, allowing
Mexico use orange and lime peel as the main pectin's the formation of oil/water emulsion (Verkempinck et al.,
raw materials, while current production in Germany 2018). Short-term stability due to bridging flocculation
mainly relies on apple pomace (Ciriminna et al., 2016). was also reported. Similarly, pectin from tomato and
broccoli extract was investigated for emulsifying and
The extraction of pectin from plant sources involves stabilizing properties (Santiago et al., 2018). The pectin
using water, acid, base, or enzyme in combination with extracted from broccoli was observed to have better
heat, microwave, ultrasound, or pulse electric field. emulsification capacity. They attributed it to its higher
Among these methods, ultrasound technology was protein content and possibly high amount of acetyl
chosen for this study because of the simplicity of groups that can provide better steric stabilization.
operation and extraction efficiency. In terms of intrinsic Additionally, sizes of emulsion droplets at various pH
benefits, ultrasound has shown more effective mixing, were monitored for 30 days, and they concluded that the
faster energy and mass transfer, lower thermal and stability of pectin-stabilized emulsion was highest at the
concentration gradient, lower extraction temperature, isoelectric pH of pectin. They furthered that lower
reduced equipment size, faster response to process dissociation of the pectin carboxylic groups at this pH
extraction control, faster start-up, increased production, led to less intramolecular repulsions, more compact
and elimination of process steps (Chemat et al., 2008). conformation of the adsorbed species at the oil-water
Studies on the use of ultrasound for pectin extraction interface, and better electrostatic interactions with
have already been done for orange peel (Yousuf et al., positively charged proteins.
2018), dragon fruit peel (Lin et al., 2018), tomato waste
(Grassino et al., 2015), jujube waste (Bai et al., 2015), This study aimed to increase the value of bananas by
grape pomace (Minjares-Fuentes et al., 2014), grapefruit extracting the pectin through ultrasonication and
peel (Bagherian et al., 2011; Wang, Ma, Xu et al., 2015), investigating the properties of banana pectin as an
and sisal waste (Maran and Priya, 2015). emulsifier. This study valorized 'saba' banana peel by
extracting pectin, a high-value food ingredient, using
In the food industry, pectin is mainly used as a ultrasound technology. Furthermore, physicochemical
gelling agent in products like jams and jellies. Due to its characterization of ultrasound-extracted pectin was
structural characteristics, pectin is also used as an performed, and a rheology study was conducted to
emulsifier or a stabilizer for emulsion products (Freitas evaluate UEP's effect on emulsions.
et al., 2021). Emulsifiers are surface-active materials that
can interact with the oil and water phases (Dickinson,
2. Materials and methods
2009). The hydrophilic portion of the emulsifier is
exposed to the aqueous phase, while the hydrophobic 2.1 Raw materials
part is in the non-polar phase. The most common Peels from unripe matured 'saba' bananas were
emulsifiers are low-molecular-weight compounds, obtained from a banana chips processing plant in Santa
typically consisting of a small hydrophilic head group Cruz, Marinduque, Philippines. It was sliced into small
and one or more hydrocarbon chains. The other type of pieces (approximately 2×2 cm2) and then oven-dried at
emulsifier is the macromolecular polymer interacting 50°C until constant weight. The dried banana peel was
with the protein component of the food (Dickinson, powdered using a countertop blender (Oster®, Classic
2009). The polymers, in sufficient concentration, form a Series Accurate Blend™, Boca Raton, Florida) and
network that can effectively trap the emulsion droplets to stored in a desiccator at room temperature. Chemicals in
prevent them from flocculating. Meanwhile, the protein analytical reagent grade were purchased from Fisher
component of the emulsion is adsorbed at the interphase, Scientific, USA.
creating a highly viscoelastic interfacial film that
2.2 Ultrasound-extraction of pectin
stabilizes the emulsions (Corredig, 2009). As a result, the
emulsion can maintain its homogeneous appearance and The protocol for the extraction of pectin from 'saba'
consistency of texture. Emulsions can also be stabilized banana peel using ultrasound technology was adapted
by particles such as silica, natural clays, and polymers. from Maran and Priya (2015) with slight modifications.
These are called Pickering emulsions, whose primary Approximately 10% (w/v) of powdered banana peel in
characteristic is the particle's affinity for both oil and 0.003 M sulfuric acid was sonicated (Sonics,
water. Pickering emulsion was named after Percival VibraCell™, Newton, Connecticut) at a 60% pressure

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 Rivadeneira et al. / Food Research 7 (1) (2023) 211 - 223 213

amplitude for 20 mins in a cold-water bath (10-15°C). methoxyl group multiplied by 100.
The sonicated mixture was filtered with a Miracloth
(Calbiochem®, Millipore Corp., Billerica, 2.3.3 Total pectic content

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Massachusetts), and the resulting filtrate was centrifuged The total pectic content was measured following the
(Thermo Fisher Scientific, Sorvall Lynx 6000, Osterode, method described by Blumenkrantz and Asboe-Hansen
Germany) at 15,000×g for 15 mins. The supernatant was (1973). A 7.5 mg sample was weighed into a test tube
filtered with a Miracloth to remove any remaining containing a magnetic stir bar. While in an ice bath (10-
residue. The pooled filtrate volume was measured, and 15°C), 5 mL cold H2SO4 was added, and the mixture was
the pectin was precipitated by adding an equal volume of stirred to allow complete solubilization of the material. A
95% ethanol. The mixture was centrifuged, and the total of 1.25 mL of deionized water was then added
precipitate was collected by filtration using Miracloth. (dropwise), followed by continuous stirring for 5 mins.
Pectin was subjected to freeze-drying at −80°C and Another 1.25 mL of deionized H2O was added
0.004 mBar for 48 hrs (Labconco, FreeZone™, Kansas (dropwise), followed by constant stirring for 5 mins. The
City, Missouri). sample was filtered with a 0.45 μm syringe filter. The
2.3 Physicochemical characterization of ultrasound- filtered sample was diluted to 1:10 using deionized
extracted pectin and low-methoxy pectin water.

The pectin samples were characterized A 1 mL of the diluted sample/standard was drawn
physicochemically to obtain the equivalent weight, out and transferred to a test tube in an ice bath (10-15°
methoxyl content, total pectic content, protein, ash, and C). Approximately 6 mL of cold H2SO4/sodium
interfacial tension. In addition, the samples were tetraborate (4.767 g sodium tetraborate in 1 L of H 2SO4)
subjected to Fourier-transform infrared (FTIR) was added to the sample. It was mixed thoroughly and
spectroscopy to identify the major functional groups and was allowed to cool in an ice bath (10-15°C). Tubes
to atomic force microscopy (AFM) to visualize the were boiled for 5 mins at 100°C, then placed
topography of the structures. The analyses were done in immediately in an ice bath (10-15°C). One-tenths
triplicates, and for the quantitative analyses, the average millilitres of 0.0125 M m-hydroxydiphenyl (0.15% in
values were reported and compared. 0.5% NaOH - stored at 4°C in a container wrapped with
aluminium foil) was added. The sample was repeatedly
2.3.1 Equivalent weight mixed in a vortex until it reached room temperature. The
absorbance of the sample was read (Thermo Scientific,
The equivalent weight was measured using Evolution 201, Madison, Wisconsin) at 520 nm against a
Rangana's method (Rangana, 1995). A 0.5 g sample was blank after 20 mins. For the standard calibration curve, 0
weighed into a 250-mL conical flask and mixed with 5 to 100 μg/mL of galacturonic acid in deionized H 2O was
mL 95% ethanol. Approximately 1 g of NaCl, 100 mL of used. For blank, 0.1 mL of 0.5% NaOH, instead of m-
deionized H2O, and six drops of phenol red were added hydroxydiphenyl, was added. The concentration of the
to the mixture. The mixture was titrated with 0.1 N total pectic substance, in terms of galacturonic acid, was
NaOH to the purple endpoint. Equivalent weight was derived from the standard curve.
calculated using the formula:
(1) 2.3.4 Protein

2.3.2 Methoxyl content Protein was measured using the protocols described
by Walker (2002). Solution A was prepared by
The methoxyl content was also measured by dissolving 1 g sodium bicinchoninate (BCA), 2 g
Rangana's method (Rangana, 1995) with modification. Na2CO3, 0.16 g sodium tartrate, 0.4 g NaOH, and 0.95 g
To the neutral solution (which contained 0.5 g of pectic NaHCO3 in 80 mL deionized H2O. The solution was
substance), after the determination of equivalent weight, adjusted to pH 11.25 using 10 M NaOH and then diluted
25 mL of 0.25 N NaOH was added. The solution was to 100 mL using deionized H2O. Solution B was
shaken manually for 30 s and set for 30 mins at room prepared by dissolving 0.4 g CuSO 4.5H2O in 8 mL
temperature in a flask with a stopper. A 25 mL portion of deionized water. The solution was then diluted to 10 mL.
0.25 N HCl was added, and the resulting solution was Solution C was made by combining solutions A and B at
titrated with 0.1 N NaOH until the purple endpoint. The a 50:1 (v/v) ratio. Approximately 2 mL of solution C
methoxyl content was calculated using the formula: were added to a 0.1 ml sample, mixed using a vortex,
(2) and then incubated at 60°C using a water bath (SI
Analytics, Weilheim, Germany) for 15 mins. It was then
Factor 3.1 is the converted equivalent weight of the cooled in an ice bath (10-15°C) to reach room

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 214 Rivadeneira et al. / Food Research 7 (1) (2023) 211 - 223

temperature. The absorbance was read against a blank Around 12.5 mg of UEP was mixed with 250 mg fine
(0.1 mL deionized water in place of the sample) at 562 potassium bromide. The mixture was pulverized
nm. For the standard, 0 to 60 μg of bovine serum manually using a mortar and pestle and put into a pellet-
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albumin (BSA) was used. The concentration of the forming die. A force was applied manually to form
protein, in terms of BSA, was derived from the standard transparent pellets. For background measurement, a
curve. pellet holder containing KBr (without pectin) was
inserted into the sample chamber (Thermo Scientific,
2.3.5 Ash Nicolet NEXUS 670 FTIR, Madison, Wisconsin).
The ash content was measured following the Infrared radiation (400-4000 cm-1) was bombarded to the
protocols described by the Association of Official sample at a resolution of 4 cm-1 with data spacing every
Analytical Chemists (AOAC) (2000). The evaporating 1.928 cm-1 for 64 scans. The same procedure was done
dish containing the sample was ignited in a furnace for the potassium bromide with pectin. The resulting
(Thermolyne, FB 1400, Dubuque, Iowa) for 5 hrs at 550° spectra were used to identify relevant peaks. LMP was
C. The ignited sample was removed from the furnace, also analyzed for comparison.
cooled in a desiccator, and then weighed. The sample 2.3.8 Atomic force microscopy of ultrasound-
was re-ignited for 30 mins. This procedure was repeated extracted pectin and low-methoxy pectin
until a constant weight was obtained. The ash content
The sample for the atomic force microscopy was
(3)
prepared following the protocols described by Li et al.
was calculated using the formula: (2019). UEP and LMP solutions (0.1 and 1% w/v) with a
final pH of 6 were prepared. The solutions were
2.3.6 Interfacial tension centrifuged at 2,000×g for 10 mins, and the supernatant
The interfacial tension was measured following the was collected for analysis. A freshly peeled mice sheet
protocols described by Li et al. (2014). Approximately attached to an AFM sample mounting disk (previously
2% (w/v) of pectin in deionized water was prepared. rinsed with 1.0 mL water at 2,000 rpm for 40 s in a
Using the Kruss FM40 Easy Drop (Hamburg, Germany), P6700 spin coater (Specialty Coating Systems Inc.,
around 15-25 μL of the solution was dosed at 10 μL/min Indianapolis, IN) was coated with 20 µL of the
on a cuvette filled with 2.5 mL n-dodecane, which was supernatant at 2,000 rpm for 20 s. The surface was
pre-purified by passing through an alumina column and covered and dried at room temperature for two hrs. The
distilled three times. The maximum droplet formed after samples were scanned under an AFM (Bruker Corp.,
dosing was incubated in the n-dodecane solution at room Multimode VIII, Santa Barbara, California) using a
temperature (25°C) for 60 mins. The droplet was then ScanAsyst-AIR probe in the ScanAsyst-Air mode. The
illuminated using the built-in illuminator, and the scanning area was 5 µm x 5 µm, and the scan rate was 1
equipment measured the interfacial tension of the pectin Hz. The generated AFM images were flattened by the
solution. Nanoscope Analysis software (Bruker Corp., Santa
Barbara, CA).
In principle, the interfacial tension between the inner
and outer phases of the pectin solution droplet resulted in 2.4 Preparation and inspection of emulsion ultrasound-
increased pressure inside the drop. The interfacial extracted pectin and low-methoxy pectin as emulsifiers
tension was calculated using the software's built-in The preparation of emulsion was adapted from the
Laplace equation (Equation 4), which describes the protocols described by Qi et al. (2020) with slight
relationship between the pressure difference and the modifications. Four set-ups were prepared wherein one
(4)
parameter was varied in each set-up. The parameters
considered were pectin concentration, oil fraction, pH,
and ionic strength.
interfacial tension.
2.4.1 Various pectin concentrations
Where Δp = pinner – pouter , σ = interfacial tension, R1,
R2 = radii of horizontal and vertical circles of curvature. UEP and LMP (0.5, 1, 2% w/v, based on the final
emulsion volume) were dissolved separately, with
2.3.7 Fourier transform infrared spectroscopy of continuous stirring, in 0.02 M NaCl solution. The final
ultrasound-extracted pectin and low-methoxy pectin pH was adjusted to 6, using 0.1 M HCl or NaOH. The
The protocol for the FTIR spectroscopy as described solution was added to soybean oil to form a 30:70 (v/v)
by Rivadeneira et al. (2020) was adopted in this analysis. oil/water mixture.

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then added to soybean oil to form a 30:70 (v/v) oil/water

mixture. The apparent viscosity profile was recorded
2.4.2 Various oil fractions from shear rates: 2 to 200 s-1 (TA Instruments

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UEP and LMP (2% w/v) were dissolved separately, Rheometer, AR 2000, New Castle, Delaware). Storage
with continuous stirring, in 0.02 M NaCl solution. The modulus (G') was also measured from 0.1 to 100 Hz at
final pH was adjusted to 6, using 0.1 M HCl or NaOH. 25°C and 0.1% strain.
The solution was mixed with soybean oil such that 30, 2.6 Emulsifying property of the emulsion
40, and 50% of oil fractions were achieved.
2.6.1 In vitro digestion of emulsion (oral phase)
2.4.3 Various pH A 30:70 (v/v) emulsion was prepared following the
procedure in 2.4.2. Approximately 14 mL of this sample
UEP and LMP (2% w/v) were dissolved separately,
was gently mixed with 0.30 mL of fresh artificial saliva
with continuous stirring, in 0.02 M NaCl solution. The
(Pickering Laboratories, Mountain View, California), 3
final pH was adjusted to 3, 4, 5, 6, and 7 using 0.1 M
mM CaCl2, 6 mM K2HPO4·3H2O, 6.4 mM KCl, 15 mM
HCl or NaOH. The solution was added to soybean oil to
NaCl, 62 mM NaHCO3, 0.005% (w/v) mucin-type II
form a 30:70 (v/v) oil/water mixture.
from the porcine stomach, and 0.1% (w/v) α-amylase
2.4.4 Various ionic strengths type VI−B for 10 seconds (Borreani et al., 2017). The
emulsion was viewed under a 40×objective.
UEP and LMP (2% w/v) were dissolved separately,
with continuous stirring, in 0, 0.02, 0.05, 0.1, and 0.2 M 2.6.2 In vitro digestion of emulsion (gastric and
NaCl solution. The final pH was adjusted to 6, using 0.1 intestinal phase)
M HCl or NaOH. It was added to soybean oil to form a
The protocols for the gastric and intestinal in vitro
30:70 (v/v) oil/water mixture.
digestion of emulsion adapted to the protocol described
2.4.5 Homogenization and ultrasonication by Borreani et al. (2017) with slight modifications. Four
and four-tenths millilitres of simulated gastric fluid
All oil-water mixtures were homogenized with a (SGF) consisting of 53 mM NaCl, 1 mM CaCl2, 5.7 mM
dispenser (Kinematica, Polytron 10-35 GT, Luzern, Na2CO3, and 14.8 mM KCl (preincubated at 37°C for 10
Switzerland) at 10,000 rpm for 2 mins. Then, the mins) were mixed with the sample from the oral phase.
resulting emulsion was subjected to ultrasonication The solution was adjusted to pH 2, added with 0.70 mg
(VCX-750, Sonics & Materials, Newton, CT) following of pepsin , and then stirred at 37°C for 120 mins (Liang
the protocol described by Wang, Ma, Xu et al. (2015) et al., 2016). The emulsion was viewed under a 40×
with slight modifications – exposing the homogenized objective.
samples to ultrasound at a pressure amplitude of 60% for
one minute. For the intestinal phase, 3 mL of bile extract solution
(4 g bile for every 100 mL of 0.5 M, pH 7 phosphate
2.4.6 Visual inspection of emulsion stability buffer) and 1.2 mL of pH 7 electrolyte mixture (0.5 mM
CaCl2, 2 mM KCl, 21 mM NaCl) were added to the
The phase separation was promoted following the
sample from the gastric phase. The solution was adjusted
protocol described by Ercelebi and Ibanoğlu (2007), with
to pH 7 and then combined with 0.50 g of pancreatin and
slight modifications. The emulsion was transferred to a
5.6 g of lipase from the porcine pancreas. The final
25-mL plastic centrifuge and centrifuged at 4000×g for 5
mixture was maintained at 37°C and pH 7 with
mins. The thickness of the formed cream layer, measured
continuous stirring for 120 mins. The emulsion was
manually using a calliper, was used to calculate the
viewed under a 40× objective.
cream fraction (thickness of the cream layer/thickness of
the whole sample). The stabilized emulsion was also 2.6.3 Fatty acid analysis
viewed under a 40× magnification of the polarized
microscope (Olympus®, BX51, Tokyo, Japan) with a Approximately 0.5 g of the sample from the
built-in digital camera (Olympus®, DP70, Tokyo, intestinal digestion was added to a 50 mL ethanol/diethyl
Japan). ether (1:1) mixture. Five drops of phenolphthalein were
added as an indicator. The sample was then titrated with
2.5 Rheology of emulsion 0.1M KOH in ethanol until the pink endpoint (which
lasted for at least 10 s) (AOCS, 2003). Acid number,
The rheological analysis of emulsion adapted the
which refers to the milligrams of KOH needed to react
protocol by Qi et al. (2020). UEP and LMP (2% w/v)
with the acid in a gram of sample, was calculated using
were dissolved separately, with continuous stirring, in
the following formula:
0.02 NaCl solution. The solution's pH was adjusted to 6,
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(5) Ultrasound extraction of pectin from other substrates

used the same parameters as 'saba' banana peel. Orange
Where C = Molarity of KOH (mmol/mL), V = Volume peel (Yousuf et al., 2018), dragon fruit peel (Lin et al.,
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of KOH used (mL), and M = Mass of the sample (g). 2018), tomato waste (Grassino et al., 2015), jujube waste
(Bai et al., 2015), grape pomace (Minjares-Fuentes et al.,
3. Results and discussion 2014), grapefruit peel (Bagherian et al., 2011; Wang,
3.1 Characteristics of ultrasound-extracted pectin and Ma, Xu et al., 2015), and sisal waste (Maran and Priya,
commercial low-methoxy pectin 2015 all used 20-30 mins exposure to ultrasonic waves.
While all these substrates used the same technology, the
The physicochemical characteristics of UEP and
studies focused on the optimization of the ultrasound-
LMP are presented in Table 1. The equivalent weight, or
extraction method and the yield of pectin. Other
the free galacturonic acid in the molecular chain of
substrates, except jujube waste, produced a higher yield
pectin (Rangana, 1995), was close to that of the LMP.
than 'saba' banana peel. However, the characteristics of
High equivalent weight indicates the high gel-forming
pectin were not studied.
activity of the two pectins (Salma et al., 2012). The
methoxyl content for UEP was within the typical values One of the proposed mechanisms for ultrasound-
for LMP, which ranges from 0.1 to 7% (Featherstone, assisted extraction is the disruption of the cell structure
2015). This characteristic, referring to the methyl ester of the plant tissue upon exposure to ultrasound
group attached to the galacturonic acid chain, determines irradiation. The said effect on plant tissue increases the
the appropriate protocol for the gelation of pectin. The accessibility of the solvent to the internal plant structure,
purity of UEP from 'saba' waste banana peel, as thereby initiating the release of the cell contents (Li et
described by the total pectic content, was within the al., 2004; Zhao et al., 2007; Supardan et al., 2012; Anese
acceptable limit (>65%) for food use set by the Joint et al., 2013; Wang, Guo, Zhang et al., 2015; Raji et al.,
FAO/WHO Expert Committee on Food Additives and 2017). The other proposed mechanism is the series of
the European Commission (Müller-Maatsch et al., 2016). expansion and compression in the medium brought about
Results also showed that UEP has a relatively higher by the ultrasound (Adetunji et al., 2017). The expansion
protein content, providing a potential synergistic effect pulls the molecules apart while the compression puts
on some food applications, especially for emulsion them together. The series of activities creates a bubble
stabilization. The mineral content (ash) of UEP was also that grows and, subsequently, experiences collapse as the
higher than LMP. This result was expected since the negative pressure exerted exceeds the local tensile
latter has higher purity. Lastly, the interfacial tension of strength of the liquid (Luque-García and Luque De
UEP was lower than LMP, which translates to higher Castro, 2003).
interfacial activities and higher stability of emulsion
(Kyomugasho, 2015). For the FTIR (Figure 2), bands from 1800 to 1500
-1
Table 1. Physicochemical characteristics of ultrasound-
cm contain the relevant functional groups for
extracted pectin (UEP) and commercial low-methoxy pectin characterizing pectin (Manrique and Lajolo, 2002). The
(LMP). spectra for UEP and LMP exhibited absorption peaks at
around 1630 - 1600 cm-1, which accounts for the
Characteristics (dry basis) UEP LMP stretching of the carboxylic acid group of the ester
Equivalent weight, g/eq 500 400 carbonyl group. A peak at around 1740 cm -1, which
Methoxyl content, % 3.18±0.1 7.61±0.1 describes the stretching of the C=O bond of the ester
Total pectic content, % 69.23±0.6 93.71±1.0 carbonyl group, was also present in LMP but not in the
Total protein, % 8.47±0.2 1.94±0.2 UEP. These findings confirmed the result of the
Ash, % 5.32±0.2 1.90±0.2 characterization where higher methoxyl content was
Interfacial tension, mN•m-1 35.40±0.5 43.59±0.2 observed for LMP.
Values are presented as mean±SD of triplicates.
AFM can be used to picture an individual pectin
Relative to an acid-extraction method (Castillo-Israel molecule and its aggregation. Figure 3 shows that both
et al., 2015), the use of ultrasound technology in UEP and LMP, measured at 5µm × 5µm, are
extracting pectin from 'saba' banana peel resulted in a combinations of elongated single polymers and
higher-purity product. A similar trend was observed aggregates. For higher concentrations (1%), UEP's
upon comparison with microwave-assisted extraction maximum height was higher than that of LMP, while the
(Rivadeneira et al., 2020) of pectin from the same opposite trend was observed at a lower concentration
substrate. Hence, ultrasound is the most effective method (0.1%). The height difference was minimal for the latter
for extracting pectin from the 'saba' banana peel in terms concentration. At high concentrations (1%), it can be
of the purity of the product.
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 Rivadeneira et al. / Food Research 7 (1) (2023) 211 - 223 217

said that some chains of the UEP overlapped, as shown said functional groups for interfacial interaction. A
by the differences in the heights within the scanned area consensus is that protein acts as an anchor between
(Adams et al., 2003). pectin and the formed oil droplet (Mengual, 1999;

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Funami et al., 2007).

Figure 2. FTIR spectra of ultrasound-extracted pectin (UEP)

from 'saba' waste banana peel and commercial low-methoxy
pectin (LMP).

Figure 4. Appearance of stabilized oil/water emulsion using

ultrasound-extracted pectin (UEP) from 'saba' waste banana
peel and commercial low-methoxy pectin (LMP) at different
conditions.

Figure 3. Atomic force microscopic images of ultrasound-

extracted pectin (UEP) from 'saba' waste banana peel and
commercial low-methoxy pectin (LMP) at concentrations of
0.1% and 1% (w/v). The size of the images is 5 µm × 5µm.

3.2 Phase separation of the emulsion

Based on Figure 4, both UEP and LMP were able to

produce a stable emulsion. This was displayed by the
formed cream layer on top of the mixture after
centrifugation, which is used to accelerate the
destabilization of the emulsion. Results (Figure 5) also Figure 5. Effect of pectin concentration, oil fraction, pH, and
showed that increasing the concentration of both UEP ionic strength on the cream fraction of oil/water emulsion
and LMP increased the volume of the stabilized stabilized by ultrasound-extracted pectin from ‘saba’ waste
emulsion. A significant difference (P<0.05) was banana peel (UEP) and commercial low-methoxy pectin
observed between the two pectins at 0.5%. The (LMP) (A: 30% oil, pH 6, 0.02 M NaCl; B: 2% pectin, pH 6,
emulsifying property of pectin can be attributed to its 0.02 M NaCl; C: 2% pectin, 30% oil, 0.02M NaCl; D: 2%
covalently bound proteins and carbohydrate moiety pectin, 30% oil, pH 6). (Grouping information based on
Tukey's Method at 95% confidence level).
(Funami et al., 2007; Schmidt et al., 2015). In this study,
higher protein content means maximum exposure of the For emulsions with UEP as an emulsifier, increasing

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the ratio of the oil component resulted in a significantly emulsifier, an increase in the volume of the stabilized
higher (P<0.05) volume of the stable emulsion. This emulsion was observed for UEP at higher NaCl
trend can be validated by the size of the emulsion concentrations. The increase in ionic strength increases
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droplets, as seen in Figure 6. Smaller droplets were the electrostatic shielding effect of Na+, resulting in the
formed at higher oil fractions; hence, the higher stability. reduction of both the charge around the emulsion droplet
The same trend was also observed in a recent study and the repulsion between pectin molecules (Zhuang et
involving multiple lights scattering to measure emulsion al., 2019). Like the effect of pH near the isoelectric pH,
stability. They found out that the stability of emulsions the pectin molecules can pack on the surface droplets
increases as the oil content of emulsions increases with more coverage, hence more stability. A similar
(Mengual, 1999). effect of ionic strength on stability was reported using
pomegranate peel pectin (Zhuang et al., 2019).

In general, UEP produced a higher volume of

stabilized emulsion than the LMP. The presence of a
higher concentration of protein in UEP may be
responsible for improved emulsion stability. The
adsorption of protein at the oil/water interface is
followed by protein unfolding, which promotes the
interactions and surface tension reduction (Dickinson
and McClements, 1995). Studies by Youssef and Barbut
(2009) on the effect of protein level on the stability of
the emulsion and that of Loi et al. (2019) presented the
same trend. That is, protein contributes to the stability of
an emulsion. They furthered that increasing the protein
composition in an emulsion improves the latter's
stability.

The appearance of droplets is presented in Figure 5

to explain the behaviour of the emulsions further.
Figure 6. Effect of pectin concentration, oil fraction, pH, and Emulsions are considered unstable when the droplets can
ionic strength on the microscopic appearance of oil/water fuse to form bigger and bigger droplets. In contrast,
emulsion stabilized by ultrasound-extracted pectin from small droplet sizes indicate more stability (Oyeneyin,
‘saba’ waste banana peel (UEP) and a commercial low- 2015). Minimal changes in the size of small droplets
methoxy pectin (LMP) (A: 30% oil, pH 6, 0.02 M NaCl; suggest that the interfacial attraction between the oil
B:2% pectin, pH 6, 0.02 M NaCl; C: 2% pectin, 30% oil, droplets is strong due to the pectin polymer forming a
0.02M NaCl; D: 2% pectin, 30% oil, pH 6).
fine film coating around individual oil droplets
Varied pH also resulted in differences in the cream (Maphosa and Jideani, 2018). Upon centrifugation, the
fraction. The volume of the stabilized emulsion produced emulsions' microscopic appearance showed that smaller
by both UEP and LMP at pH 3 and 4 were higher than at droplets were formed for all parameters when UEP was
pH 5 to 7. In a related study, Akhtar et al. (2001) used as an emulsifier; hence, a more stable emulsion.
compared citrus pectin-stabilized emulsions at pH 4.7 This finding supports the calculated cream fractions of
and 7 and observed that emulsions were more stable at the emulsions, where UEP has higher values than LMP.
the lower pH. According to Nakauma et al. (2008), It was also observed that changes in the size of droplets
emulsions flocculate at the isoelectric point of the were most visible for various oil fractions in UEP
stabilizer's carbohydrate moiety due to reduced emulsion. As the oil fraction increased, smaller droplets
repulsion. Their study utilizing sugar beet pectin were formed, and a higher cream fraction was recorded,
decreased emulsion stability at pH above the pectin's hence, increasing stability.
isoelectric point. The same trend was also observed in 3.3 Rheological property of the emulsion
the sizes of the emulsion droplets, which are smaller at
the isoelectric pH of the pectin. Using UEP and LMP as emulsifiers, both emulsions
exhibited a non-ideal liquid with shear-thinning
The presence of NaCl also affected the stabilization behaviour wherein the apparent viscosity decreases with
of the emulsion. While the amount of NaCl resulted in a increasing shear rate (Figure 7). Between the two, UEP
non-significant (p<0.05) change in the volume of produced an emulsion with higher viscosity. This finding
stabilized emulsion when LMP was used as an can be explained by the difference in the stabilizers' level

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 Rivadeneira et al. / Food Research 7 (1) (2023) 211 - 223 219

FULL PAPER
Figure 7. Rheology of oil/water emulsion stabilized by ultrasound-extracted pectin (UEP) from 'saba' waste banana peel (UEP)
and commercial low-methoxy pectin (LMP) (2% pectin, 30% oil, pH 6, 0.02M NaCl).
of esterification, with LMP doubling the methoxyl calorie emulsion-based functional food products.
content of UEP. A study comparing the effect of the
citrus pectin's degree of esterification (DE) on the
intrinsic viscosity showed that increasing the DE
decreases the latter (Morris et al., 2000). They furthered
that pectin with lower DE is more rigid. It may also be
attributed to the charge distribution along the molecule
(Yoo et al., 2006). Lower DE translates to lower surface
charge, less repulsion, and, thus, higher viscosity of the
continuous phase. Lastly, based on the microscopic
appearances of the respective emulsions, UEP produced
smaller droplets, which decreased the average distance
between droplets, leading to an increase in both the
hydrodynamic interaction and the viscosity (Pal, 1996).

3.4 In vitro digestion of emulsion

Digestion of lipids consists of an interfacial reaction

since it depends on the lipase adsorption onto the oil
droplet surface (Torcello-Gómez et al., 2011). Therefore,
incorporating it in an emulsion can improve digestion
since a larger surface area will be available for lipase
adsorption.
Figure 8. Appearance of oil/water emulsion stabilized by
Results (Figure 8) showed that both emulsions using ultrasound-extracted pectin (UEP) from 'saba' waste banana
UEP and LMP were found to be stable in all stages of in peel (UEP) and commercial low-methoxy pectin (LMP) at
different stages of digestion.
vitro digestion. While UEP provided a more stable
emulsion before digestion, subjecting both emulsions to
oral, gastric, and intestinal digestion did not affect the
original stability.

In the final phase of digestion, hydrolysis of the

ingested lipid generates free fatty acid (FFA) and
diacylglycerols (Pafumi et al., 2002). Figure 9 shows a
drastic increase in fatty acid in the first hour of intestinal
digestion and a gradual increase in the remaining hour.
Between the two emulsifiers, UEP resulted in a lower
concentration of fatty acid, indicating its ability to inhibit
lipid digestion.

Overall, the observed stability of emulsion

Figure 9. Free fatty acid concentration during the intestinal
throughout the in vitro digestion and the lower fatty acid digestion of oil/water emulsion stabilized by ultrasound-
concentration in the intestinal phase suggest that the UEP extracted pectin (UEP) from 'saba' waste banana peel (UEP)
can be used in inhibiting lipid digestibility and can and commercial low-methoxy pectin (LMP) (2% pectin, 30%
potentially be used in the development of reduced- oil, pH 6, 0.02 M NaCl).

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 220 Rivadeneira et al. / Food Research 7 (1) (2023) 211 - 223

4. Conclusion Food Chemistry, 136(2), 458–463. https://

Using UEP from 'saba' waste banana peel as an doi.org/10.1016/j.foodchem.2012.08.013
FULL PAPER

emulsifier resulted in a more stable emulsion than an Association of Official Analytical Chemists (AOAC).
LMP. Despite having lower purity, the synergistic effects (2000). Official methods of analysis, 17th ed. USA:
of the protein and pectin established a lower interfacial AOAC
interaction with oil and water. Higher pectin and ion Bai, F., Wang, J. and Guo, J. (2015). Optimization for
concentration, higher oil fraction, and pH close to ultrasound-microwave assisted extraction of pectin
pectin's isoelectric pH were the conditions that increased from jujube waste using response surface
the volume of the stabilized emulsion. For both UEP and methodology. Advance Journal of Food Science and
LMP, in vitro digestion did not affect the stability of the Technology, 7(3), 144–153. https://doi.org/10.19026/
emulsion, with the former performing better in inhibiting ajfst.7.1283
lipid digestion. The production of UEP, considering the
Bagherian, H., Ashtiani, F.Z., Fouladitajar, A. and
results of this study, has increased the value of the
Mohtashamy, M. (2011). Comparisons between
Philippine 'saba' banana. The utilization of waste banana
conventional, microwave- and ultrasound-assisted
peel as a source of high-value food ingredients is another
methods for extraction of pectin from grapefruit.
leap towards zero-waste food production.
Chemical Engineering and Processing: Process
Intensification, 50(11–12), 1237–1243. https://
Conflict of interest doi.org/10.1016/j.cep.2011.08.002
The authors declare no conflict of interest. Blumenkrantz, N. and Asboe-Hansen, G. (1973). New
method for quantitative determination of uronic
Acknowledgements acids. Analytical Biochemistry, 54(2), 484–489.
https://doi.org/10.1016/0003-2697(73)90377-1
This study was funded by the Department of Science
and Technology's Accelerated Science and Technology Bon, S.A. (2014). The phenomenon of Pickering
Human Resource and Development Program (DOST- stabilization: a basic introduction, In Ngai, T. and
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