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Letter

pubs.acs.org/journal/ascecg

Lemon Juice Based Extraction of Pectin from Mango Peels: Waste to


Wealth by Sustainable Approaches
Jhumur Banerjee,†,‡,§ R. Vijayaraghavan,§ Amit Arora,† Douglas R. MacFarlane,*,§ and Antonio F. Patti*,§

Centre for Technology Alternatives for Rural Areas (CTARA), Indian Institute of Technology Bombay, Powai, Mumbai 400076,
India

IITB Monash Research Academy, Indian Institute of Technology Bombay, Powai, Mumbai 400076, India
§
School of Chemistry, Green Chemical Futures, Monash University, Wellington Road, Clayton, Victoria 3800, Australia
*
S Supporting Information

ABSTRACT: Valorization of mango peels to recover pectin


has the potential to increase the economic viability of a
biorefinery utilizing this waste resource. Replacement of
conventional mineral acids used for extraction of pectin with
natural food grade acids would assist in making the process
more environmentally friendly and safe for food applications.
In this work, we have evaluated the effect of a natural
acidifying agent, lemon juice, in combination with sonication,
on pectin extraction. Sonication was used for 20 min at 80 °C,
compared to the conventional process which involves boiling
for 150 min, thus improving the energy cost of the process considerably. More than 26%w/w of the mass of dried mango peel
was extracted as pectin; this was classified as low methoxyl pectin (degree of esterification ≤ 50%). Pectin having this DE is of
importance in low calorie food products as a nutraceutical additive.
KEYWORDS: Mango peels, Pectin, Sonication, Valorization

■ INTRODUCTION
Pectin is a structural carbohydrate that is found abundantly in
5−10%. The majority of the carbohydrates were found to be
soluble dietary fiber such as pectin.16 Compared to other
plant processing waste.1 It provides strength to plant tissues commercial sources12 such as apple pomace (10−15% pectin),
citrus peel (25−35% pectin), sugar beet (10−20% pectin) and
and helps in adherence. Many fruits contain a significant
sunflower (15−25% pectin), the quantity of pectin in mango
quantity of pectin in their peels. The application of pectin as a
peels varies from 20 to 30% of total peel weight.9 Techniques
gelling and thickening agent in food,2 as soluble dietary fiber,3
such as stirring with hot acids and microwave assisted hot acid
as a drug delivery carrier, as a film-forming polymer and
treatment have been reported to yield 9−26% (w/w) of pectin
recently as a prebiotic oligosaccharide has generated significant
from the mango peels.17−19 These previously reported methods
interest in efficient and economical methods for its
include treatment of the peels with mineral acids (hydrochloric
production.4−7 Recent studies have also indicated its role in
acid, nitric acid, sulfuric acid) at high temperature for long
prevention of pathogenic adhesion,8 chelation of heavy metals durations.17,20 The resultant byproduct from such industrial
inside the body and repair of connective tissues.6 processes is thus rich in mineral acids, which ultimately creates
Commercially, pectin is produced from citrus peels and apple a waste stream that needs to be treated and discarded.21,22
pomace.9 The cost of pectin10 used in industries is typically > Other methods, in which citric acid is used as a chelating
US$11/kg. On the other hand, because many other pectin rich agent,2 need to employ large quantities of this relatively
fruit peels and pomace are waste produced from food expensive acid, because of its weakly acidic nature.23 In all cases,
industries5,11 and typically have value < $0.1/kg, a recovery the result is a relatively expensive process that therefore creates
approach for pectin from this waste could provide new a high cost product.
commercial sources from material that is otherwise discarded. The use of natural sources of the acidifying agent such as
Thus, in a recent report, pectin was shown to be a valuable date juice and lemon juice has been reported24,25 in limited
product component in the design of a biorefinery.12
Mango peels consist of approximately 20−40% of the total
Special Issue: Building on 25 Years of Green Chemistry and
mango processing waste (by weight) generated in indus-
Engineering for a Sustainable Future
tries.13,14 The other major components, as determined
experimentally15 on an Indian variety of mango peels, were as Received: June 14, 2016
follows: cellulose, 15−18%; hemicellulose, 5−11%; lignin, 9− Revised: August 31, 2016
12%; pectin, 20−35%; proteins, 6−10%; ash, 2%; extractives, Published: September 5, 2016

© 2016 American Chemical Society 5915 DOI: 10.1021/acssuschemeng.6b01342


ACS Sustainable Chem. Eng. 2016, 4, 5915−5920
ACS Sustainable Chemistry & Engineering Letter

Figure 1. (a) Pectin yield from sonication assisted extraction (30 kHz, 20 min) for different temperatures (40, 50, 60 and 80 °C) (water-peel, pH
4.0; lemon treated, pH 2.5, time: 20 min). (b) Comparison between pectin yield from conventional vs sonication assisted extraction. (T = 80 °C, t =
150 min for conventional and 20 min for sonication. Means that do not share a common label are significantly different.)

studies. Conventional boiling for duration of 90−180 min was juice as acidifying agent is compared with mineral acid (HCl)
carried out and found to yield significant quantities of pectin and also with an aqueous medium with no added acid. Lemon
from passion fruit25 and lemon peels.24 The advantages of using juice mainly consists of citric acid (45−63 g/L), malic acid (1−
such agents is considered to be the reduction of mineral acid 7 g/L), ascorbic acid (0.2−0.4 g/L) and tartaric acid (0.05−0.1
waste as byproduct and the improved nature of the final g/L).32,33 To the best of our knowledge, lemon juice as an
product due to the food grade nature of the extracting solvent. acidifying agent for pectin extraction using sonication has not
Recently, sonication based methods were shown to be often been reported previously and provides an advantage over
greener, efficient and more economical compared to heating for existing methods due to its intrinsic biocompatibility and
extraction processes.26 15−17% yield of pectins was obtained acceptability for food grade applications.
from tomato peels27 within 15 min of sonication compared to
15−21% yield in 24 h through the conventional heating
process. Sonication of Artocarpus peels for 24 min at 60 °C
■ EXPERIMENTAL SECTION
Kensington variety mangoes and Lisbon variety lemons were
recovered 14.5% of pectin.28 In another recent study on purchased from a local market in Clayton, Melbourne, and mango
grapefruit peels, a sonication method was compared with the peels were separated from the flesh. The obtained peels were washed
with water to remove adhered pulp and debris. The washed peels were
conventional process in terms of the pectin characterization. A
blanched in hot water at 85 °C for 5 min. Washed peels were dried in a
lowering in degree of esterification was observed in the case of hot air food drier at 55 °C with an air flow of 2 m/s until constant
sonication and the pectins obtained were found to be superior weight was obtained. The dried peels were ground in a food grinder to
to that of the conventionally extracted pectins in terms of obtain a powder. A fine powder of particle size <500 μm was obtained,
thermal stability and bioactivity.29 The increased efficiency in packed and stored in airtight bags at 4 °C until further use. Lemon
most of these studies was correlated with a several fold decrease juice obtained from the fruit was filtered and centrifuged to remove
in extraction time. A significant disruption in vegetal tissues was any solids. An aliquot was mixed with 1:1 volume of ethanol to detect
observed due to the sonication which helped in release of the for any pectin in the juice that was subtracted from the final yield of
pectin obtained from the mango peel mixtures. All reagents and
pectin.30,31 chemicals used were of analytical grade. Pectin standard was purchased
In this work, we demonstrate a method for the extraction of from Sigma-Aldrich (USA).
high pectin yields from mango peels using a combination of Pectin Extraction. Because mango peels are rich in valuable
lemon juice as acidifying agent and a sonication process. Lemon polyphenols, we extracted the polyphenols first, followed by the use of

5916 DOI: 10.1021/acssuschemeng.6b01342


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the residual powder for pectin extraction. The extractive free powder
of mango peels was prepared using Soxhlet extraction apparatus.13
Hexane was used as solvent to remove wax, lipids and pigments (2.4 ±
■ RESULTS AND DISCUSSION
The yields from sonication assisted extraction for aqueous and
1.2%) followed by 4 h of extraction (until the percolated extract lemon juice acidified mixtures at different temperatures are
became colorless) with ethanol to remove further any proteins, free shown in Figure 1a. The yield of pectin was found to be
sugars and polar extractives such as polyphenols. The ethanol was affected by acidity. In general, the yield of pectin increases with
recovered by distillation and further used in separating pectins. The acidity until pH 2.0, below which the likelihood of hydrolysis of
yield of ethanol extract was found to be 15.2 ± 1.5% of total powder pectin is high.39 In our study, an increase in yield with lemon
weight. juice, as compared to water, is thus justified on the basis of
For the pectin extraction, a slurry of the extractive free mango peel previous reports.37,39,40 Acidity helps in release of protopectin
powder (1 g) was prepared in distilled water (20 mL)11 with and from the native structure of the cell wall in which protopectin is
without the addition of acids/lemon juice. The natural pH of mango often bound between the hemicellulose, lignin and cellulose
peel solution at solid−liquid ratio of 1:20 was found to be 4.0, which layers.41
was used as a control. The pH of the solution was kept at 2.5 for all the The role of temperature along with sonication was observed
experiments except the control23 and accordingly a specific quantity of clearly in the case of lemon juice acidification. The highest yield
lemon juice or mineral acid was used. HCl was chosen as mineral acid was obtained at 80 °C (26.6% w/w). This yield was 1.7 times
based on a previous study.17 that of 60 °C. This is in agreement with previous studies in
Pectin extraction was compared between the two methods:
which increased temperature was shown to enhance the
sonication and conventional boiling with no acid (water, pH 4.0),
mineral acid (pH 2.5) and lemon juice (pH 2.5). On the basis of
solubility of protopectin. For this reason, 80 °C was chosen
preliminary experiments, we found that maximum pectin was extracted as the optimum temperature for the next set of sonication
at 80 °C through sonication in the presence of lemon juice (Figure based extractions. Figure 1b shows the difference between the
1a). Thus, 80 °C was chosen for subsequent extraction experiments. In conventional extraction and sonication based extraction. With
the second step, a conventional process (boiling with mineral acid at the help of statistical analysis (ANOVA), it was found that the
80 °C for 150 min) was compared with sonication (Elmasonic S60H, lowest yields were obtained when no acid was added (p < 0.05).
37 kHz, 550 W; conditions: 20 min,34 80 °C). The process flowchart Among the acidified media, lemon juice acidification with
for the comparative study between the two methods is shown in sonication and conventional extraction was found to yield a
Figure 1b along with results. The quantity of pectin in filtered and similar quantity of pectin (p < 0.05). In the case of mineral acid,
centrifuged lemon juice was found to be negligible and was determined the conventional method was found to yield more pectin than
to avoid any yield due to lemon juice. In the case of sonication, sonication, which may be due to effect of pH and temperature
temperature variations were minimized by regulating the temperature on the hydrolysis of pectin. The improved yield with sonication
of the water. After the treatment the flasks were allowed to cool to may be due to the cavitation effect,26 which aids in disruption of
room temperature. The obtained viscous mass was centrifuged at 6000 cell wall structure and easy separation of protopectin is
rpm for 15 min to separate the solids. The supernatant was collected, achieved. The natural extracting ability of lemon juice may also
and a 1:1 volume of absolute alcohol was added to precipitate pectic be linked to presence of the combination of different acids.24
polysaccharide. The solutions were stored at 4 °C overnight. The Also, the presence of salts in the juice may form a buffer that
precipitated gels were again centrifuged (6000 rpm, 15 min), and the
prevents substantial change of pH over the process duration
pellet was collected and washed three times with alcohol−water (1:1)
mixture to remove any adhering sugars or polyphenolics. The obtained
and this may be advantageous compared to the use of pure
pellet was filtered and dried at 45 °C to constant weight. The yield was organic acids such as citric acid (pKa1 for citric acid = 3.1, hence
calculated with respect to the weight of initial powder weight. pH = 2.5 is within the buffer region of this acid).25
Characterization of Pectin. Attenuated total reflectance Fourier The characterizations of commercial, conventional and
transform infrared spectroscopy (ATR/FT-IR) was performed on lemon extracted pectins through FTIR are shown in ESI
dried pectin samples using a PerkinElmer Cary 630 ATR/FT-IR Figure 2. The characteristic bands around 3300−3500 cm−1
equipped with MicroQuant software. The recorded spectrum was an denote OH stretching. The broad absorption observed in
average of 32 scans with spectral resolution of 4 cm−1 from 500 to our samples compared to the commercial pectin may be due to
4000 cm−1.35 The background spectrum was collected, followed by the hydrogen bonding of hydroxyl groups and carboxylic acid
sample spectra collection.35 The spectrum of lemon juice extracted dimers of polysaccharide.27 The band at around 2950 cm−1 is
pectins was compared with the commercial pectin. The FTIR due to the CH stretching of galactrouronic ring. Bands
technique is also a standard industrial method for qualitative analysis around another important region that helps in the detection of
of pectins.4 The other method of characterization was determination esterified vs unesterified (methylated) functional groups is the
of the degree of esterification (DE) value. This is the percentage of fingerprint region 1750−1350 cm−1. Both the commercial
galactrouronic acids present in methyl ester form.35 The DE value sample and the extracted pectins produced an intense band
determines the gelling nature of pectin.7,36,37 Generally, pectins having around 1740 cm−1. The patterns were found to be well
DE value >50% are termed “high methoxyl pectins”, which form gels correlated to that of commercial pectin. The change in intensity
in the presence of sugar.38 Low methoxyl pectins have a DE value
of predicted −COOCH3/−COOH region may be related to
<50% and are often prepared by controlled hydrolysis of high
methoxyl pectins. Low DE pectins form gels in the presence of calcium
the lower degree of esterification in the case of sonication and
ions. The degree of esterification was determined by a titration method lemon juice extracted pectin. The band around 1600 cm−1 is
that is commonly followed in industry.36 The method was modified due to the carboxylate group. Small bands at around 1200 and
slightly in terms of the pectin weight dissolved in water (200 mg/40 1100 cm−1 may be due to the glycoside ring, CO stretching
mL instead of 500 mg/100 mL36). and OH bending.27 Bands around 650 cm−1 may be due to
Statistical Analysis. The difference between triplicate values in the presence of the pyranoid ring structure of pectin.27 Similar
yields obtained by sonication were evaluated using a one way analysis observations were reported for tomato, citrus and pumpkin
of variance (ANOVA) with Tukey test. Analysis was carried out with pectin where a high intensity band at 1749 cm−1 was correlated
the help of Minitab 16 (Minitab Inc. USA). with stretching of a methylated carboxyl (CO).42
5917 DOI: 10.1021/acssuschemeng.6b01342
ACS Sustainable Chem. Eng. 2016, 4, 5915−5920
ACS Sustainable Chemistry & Engineering Letter

Figure 2. Degree of esterification for different pectins obtained from sonication assisted extraction (80 °C, 20 min).

The DE values for extracted pectins are shown in Figure 2.


Commercial pectin has the highest DE value, followed by

*
ASSOCIATED CONTENT
S Supporting Information
mineral acid and lemon juice extracted pectins. Mineral acid
helps in extraction of pectins with a DE value of 69%, whereas The Supporting Information is available free of charge on the
sonication with lemon juice yields a lower DE value of around ACS Publications website at DOI: 10.1021/acssusche-
50%. This may be due to increased demethylation in the meng.6b01342.
presence of lemon juice with sonication. A similar observation Flowchart of the pectin extraction process and FTIR
with respect to sonication was reported for grapefruit and other spectra of pectins (PDF)


pectic materials where a lowering in DE value was observed
with increase in time and temperature.30,43 The commercial
grades of pectins generally suggested for food grade have a DE AUTHOR INFORMATION
value >60% for the gelation in the presence of sugars.44 On the Corresponding Authors
other hand, in many recent studies low DE pectin was found to *D. R. MacFarlane. Email: doug.macfarlane@monash.edu.
be useful in low calorie and low fat products, because it requires Phone: +61 3 9905 4540. Fax: +61 3 9905 4597.
calcium for gelling and little or no sugar.45 Typically, low DE *A. F. Patti. Email: tony.patti@monash.edu. Phone: +61 3 9905
pectin has higher value than the higher DE products.10 Low DE 1620. Fax: +61 3 9905 4597.
pectin was also found to be beneficial in lowering serum
Notes
cholesterol level in humans as shown in several in vivo studies.46
In a study done on mucin secretion in rats and the cultured The authors declare no competing financial interest.
HT29-MTX cell line, low DE pectins were found to increase
the secretion of mucin;46 this in turn suggested increased gut
protection and a strong intestinal barrier against infections, e.g.,
■ ACKNOWLEDGMENTS
We are grateful to the School of Chemistry, Monash University
bacterial infections. Thus, the pectin extracted from the mango for the facilities and Ministry of Food Processing Industries
peels by the methods described here may find increasing (MOFPI) under Department of Science and Technology
application as nutraceutical additives in food products. (DST) India for providing funding through the external

■ CONCLUSIONS
This initial study of the valorization of a large food processing
competitive grants program (Project ID: SERB/MOFPI/
0036/2013). D.R.M. is grateful to the Australian Research
Council for support under the Australian Laureate Fellowships
waste resource demonstrates the use of lemon juice (pH 2.5) as program.
an extraction medium for pectin from mango peels. Valor-
ization of these peels by pectin production not only adds value
to the generated waste but mango peel pectin’s qualitative
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