See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/349650925

Jackfruit seed slimy sheath, a novel source of pectin: Studies on antioxidant

activity, functional group, and structural morphology

Article  in  Carbohydrate Polymer Technologies and Applications · February 2021

DOI: 10.1016/j.carpta.2021.100054

CITATIONS READS

29 1,601

10 authors, including:

Manoj Kumar Jayashree Potkule

Central Institute for Research on Cotton Technology Institute of Chemical Technology, Mumbai
282 PUBLICATIONS   4,024 CITATIONS    13 PUBLICATIONS   371 CITATIONS   

SEE PROFILE SEE PROFILE

Maharishi Tomar Sneh Punia Bangar

Indian Grassland and Fodder Research Institute Clemson University
62 PUBLICATIONS   1,269 CITATIONS    257 PUBLICATIONS   3,954 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Innovations in starch-based edible films and coatings View project

Competitive invitation for book submission to Book Series “Cereals: Science and Processing Technology View project

All content following this page was uploaded by Manoj Kumar on 07 March 2021.

The user has requested enhancement of the downloaded file.

 Carbohydrate Polymer Technologies and Applications 2 (2021) 100054

Contents lists available at ScienceDirect

Carbohydrate Polymer Technologies and Applications

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

Jackfruit seed slimy sheath, a novel source of pectin: Studies on

antioxidant activity, functional group, and structural morphology
Manoj Kumar a,∗, Jayashree Potkule a, Maharishi Tomar b, Sneh Punia c,d, Surinder Singh e,
Sharmila Patil f, Sudha Singh g, T. Ilakiya h, Charanjit Kaur i, John F. Kennedy j
a
Chemical and Biochemical Processing Division, ICAR-Central Institute for Research on Cotton Technology, Matunga, Mumbai 400019, India
b
Division of Seed Technology, ICAR - Indian Grassland and Fodder Research Institute, Jhansi 28400, India
c
Department of Food Science and Technology, Chaudhary Devi Lal University, Sirsa, India
d
Department of Food, Nutrition, & packaging Sciences, Clemson University, Clemson, SC 29634, United States
e
Dr. S.S. Bhatnagar University Institute of Chemical Engineering and Technology, Panjab University, Chandigarh 160014, India
f
Quality Evaluation and Improvement Division, ICAR – Central Institute for Research on Cotton Technology, Matunga, Mumbai 400019, India
g
Department of Vegetable Science, University of Horticulture and Forestry, Nauni, 173230 Solan, Himachal Pradesh, India
h
Department of Vegetable Science, Tamil Nadu Agricultural University, Coimbatore 641003, Tamil Nadu, India
i
Division of Food Science and Post-Harvest Technology, ICAR - Indian Agricultural Research Institute, New Delhi 110012, India
j
Chembiotech Laboratories, Advanced Science and Technology Institute, Kyrewood House, Tenbury Wells, Worcestershire WR15 8FF, UK

a r t i c l e i n f o a b s t r a c t

Keywords: In the current study, pectin was extracted from the unexplored slimy sheath present over seed coat of jackfruit by
Jackfruit seed slimy sheath pectin (JSSP) using oxalic. Extracted jackfruit seed slimy pectin (JSSP) demonstrated higher total phenolic content (65.7 mg
Fourier-transform infrared (FTIR) spectroscope GAE/g) than commercial pectins from apple (51.3 mg GAE/g) and citrus (30.7 mg GAE/g). Antioxidant activity
analysis
using ferric reducing antioxidant power assay for JSSP was also higher (10.4 µM) compared to commercial pectins
Scanning electron microscopy (SEM)
(4.21–5.96 µM) which was attributed to co-extraction of phenolics along with JSSP. Microstructure of the JSSP
Antioxidant activity
was observed using scanning electron microscopy and it was found that smooth and flattened morphology with
irregularities in particle size of JSSP. Functional group analysis of JSSP using Fourier-transform infrared (FTIR)
spectroscopy was found similar to that of commercial apple and citrus pectins. These parameters established
jackfruit seed slimy sheath as a novel source of antioxidant rich pectin for application in functional foods and
medical field.

1. Introduction utilization can be in form of extraction of biological macromolecules in-

Jackfruit is the largest tree-borne, tropical fruit, widely cultivated
and produced in countries like India, Bangladesh, Thailand, Indone-
sia and Nepal (FAO, World Atlas, accessed on 2019). The edible
portion of jackfruit is nutritious as it is a source of various min-
erals (iron, potassium, and calcium), vitamins (A, B, C), carbohy-
drates, proteins and other phytoceuticals (APAARI, 2012). Apart from
the edible portion, a lot of work has been done on utilization of
non-edible portion viz. seeds (Akter & Haque, 2018; Nayak, Pal, &
Santra, 2015; Suryadevara, Lankapalli, Danda, Pendyala, & Katta, 2017;
Tramontin et al., 2019; Zhang, Hu, Zhu, Wu, & Tan, 2018; Zhang et al.,
2019) and peels (Govindaraj, Rajan, Hatamleh, & Munusamy, 2018;
Jancy, Shruthy, & Preetha, 2019; Sundarraj, Thottiam Vasudevan, & Sri-
ramulu, 2018) left after the extraction of edible flakes from the fruit. The
cluding starch, pectin and proteins. Among all, pectin is the most valu-
able valorized product from jackfruit waste. It is a natural additive ex-
tensively used in the food industry as a stabilizer, gelling agent, textur-
izer and pharmaceutical industry (Liu, Cao, Huang, Cai, & Yao, 2010).
The estimated consumption of pectins was 34000 metric tonnes in 2016
and its demands may soar up to 48,735 metric tonnes with a CAGR
of 3.7% by the end of 2026 (Sundarraj et al., 2018). Despite increas-
ing demand, it is becoming difficult for the food processing industries
for achieving high production of pectin from traditional sources namely
apples and citrus peels. Hence, wastes generated from the fruits and veg-
etable processing industry can act as a sustainable and potential source
of pectin while also addressing the issue of environmental pollution.

Abbreviations: JSSP, Jackfruit Seed Slimy Sheath Pectin; JSS, Jackfruit Slimy Sheath; FTIR, Fourier Transform Infrared Spectroscopy; GAE, Gallic Acid Equivalents;
SEM, Scanning Electron Microscopy; FAO, Food and Agriculture Organization; APAARI, Asia-Pacific Association of Agricultural Research Institutions; TPC, Total
Phenolic Content; DPPH, 2,2-diphenyl-1-picrylhydrazyl; KBr, Potassium bromide; AOA, Antioxidant activity; RT, Room temperature.
∗
Corresponding author.

https://doi.org/10.1016/j.carpta.2021.100054
Received 17 November 2020; Received in revised form 15 February 2021; Accepted 26 February 2021
Available online 28 February 2021
2666-8939/© 2021 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
M. Kumar, J. Potkule, M. Tomar et al.
Accordingly, slimy sheath present over the jackfruit seed coat can be
constructively utilized for the production of high-value pectin.
Commercial pectins are most commonly extracted using strong acids
but increasing concerns of consumers for the safety of the food prod-
ucts have led to exploring the new extraction ways with low chemical
residues and higher extraction efficiency. As far as there are no pub-
lished findings for the extraction of pectin from the jackfruit seed slimy
sheath layer. However, Sundarraj et al., (2018), extracted jackfruit peel
pectin using organic acid (citric, tartaric, & oxalic acid) and inorganic
acids (hydrochloric & nitric acid) and achieved the highest yield of
38.4% by using 0.05N oxalic acid (temperature = 90 °C and time = 1 h).
Therefore, in the current study oxalic acid (0.05N) was used for the ex-
traction of pectin from the unexplored slimy sheath which is present
just over the seed coat of jackfruit (for more clarification see the video
uploaded along with this research article). The pectins extracted from
the fruit wastes are usually associated with the biological activities (an-
tioxidant, antitumor, anti-inflammatory) (Ho et al., 2015; Kumar et al.,
2020), hence, the antioxidant property of JSSP was compared with ex-
isting pectic substances from the citrus and apple our study. The phe-
nolic content of the novel jackfruit seed slimy sheath pectin (JSSP) was
also evaluated to establish the correlation with the antioxidant activ-
ities. Further, the functional groups and microstructure of JSSP were
compared with the commercially available pectin with the help of FTIR
spectroscopy and SEM, respectively. This study will provide the basis
of application of novel pectin from jackfruit to be utilized in food and
medical field.
2. Materials and methods
2.1. Materials
Jackfruit flakes of ripened fruit were purchased from the local mar-
ket, Ghatkopar, Mumbai, Maharashtra, India and jackfruit seed slimy
sheath were recovered for the extraction of pectin. The pectin from ap-
ple and citrus were procured from Sigma chemicals Co. P. O. St. Louis,
USA.
2.1.1. Reagents
2,2-diphenyl-1-picrylhydrazyl, Folin-Ciocalteu reagent, D-
galacturonic acid were purchased from HiMedia (Mumbai, Maha-
rashtra, India). 2,4,6-tri(2-pyridyl)-s-trizine, gallic acid, potassium
bromide procured from Sigma-Aldrich Co. (St. Louis, USA), absolute
ethanol 99.9%, methanol, oxalic acid, sodium carbonate, ferric chloride
and other chemicals and reagents were analytical grade.
2.2. Experimental method
2.2.1. Extraction of Pectin
Pectin was extracted from undiscovered slimy sheath present over
the seed coat of jackfruit by using method given by Sundarraj et al.,
(2018) Taking 25 mL deionized water into the 50 mL of beaker and
acidified with 0.05 N oxalic acid. Jackfruit seed coat slimy sheath (JSS)
powder was prepared by first drying JSS in the hot air oven at 50 °C
for 24 h (Meta lab scientific industries, Mumbai, India). After drying
JSS was powdered using a mortar pestle and stored in an airtight con-
tainer at 4 °C. 2.5 g of JSS powder was accurately weighed and mixed
in acidified 25 mL deionized water. The mix was placed in a water bath
(Instrucare solution, Mumbai, India) at 90 °C for 1 h and 200 rpm. Ox-
alic acid extracted hot solution was filtered through muslincloth in a
100 mL beaker and allow to cool at RT. 1:2 volume of 95% ethanol was
added for precipitation of pectin and dried at 50 °C for 16 h in a hot air
oven. After drying it was powdered using a mortar pestle and stored in
an airtight container. The flow diagram of pectin extraction using acid
extraction has been presented in Fig. 1. JSSP yield on dry weight basis
was determined using the following formula,
[ ]
Pectin yield(%) = 100X P∕Bi
2
Carbohydrate Polymer Technologies and Applications 2 (2021) 100054
P: Amount of extracted jackfruit seed slimy layer pectin on a dry weight
basis.
Bi: Initial amount of jackfruit seed slimy layer powder on a dry
weight basis.
2.3. Analytical methods
The methods employed for the analysis of galacturonic acid, total
phenolic content, antioxidant activity, functional groups identification
and microstructure are as follows:
2.3.1. Galacturonic acid (GalA) content
The galacturonic acid content of JSSP, JSS powder, commercial ap-
ple and citrus pectin was determined by the carbazole and sulphuric
acid spectrophotometric method (Jin & Yang, 2019). 1 g of sample was
dissolved with distilled water at 55 °C and then diluted up to 100 mL.
1 mL of pectin solution was mixed with 6 ml of concentrated sulphuric
acid (98%, v/v) and heated in a boiling water bath for 20 min. After
cooling to RT, 0.5 mL of 0.15% carbazole solution (prepared in abso-
lute ethanol) was added, mixed with gentle stirring, and then allowed
to stand in the dark for 1.5 h. The absorbance was measured at 530 nm
against the blank reagent with D-galacturonic acid as standard.
2.3.2. Protein content
The protein content of JSSP, JSS powder, commercial apple and cit-
rus pectin was determined by Bradford method (Kruger, 2009).
2.3.3. Total phenolic content (TPC)
TPC of JSSP, JSS powder, commercial apple and citrus pectin was
determined by Asgari, Labbafi, Khodaiyan, Kazemi and Saeid Hos-
seini, (2019) method. 0.5 mL (1% w/v) sample was poured into the
10 mL test tube. Later, Folin-Ciocalteu reagent (2.5 mL, 10% v/v) added
in sodium carbonate (2 mL, 7.5% w/v). The mix was then vortexed
for 1 min and then stand at RT for 1 h. Absorbance of the sample
was recorded at 750 nm using UV 1700 spectrophotometer, Shimadzu,
Japan. Calculate TPC of JSSP, JSS powder, apple and citrus pectin and
values were expressed as mg gallic acid equivalents (GAE)/g of sample
on a dry weight basis.
2.3.4. Antioxidant activity by DPPH assay
DPPH method described by Hendel, Larous, and Belbey, (2016) was
used. 100 µl (10 mg/mL) of methanol extract solution were added into
the 5 mL 0.004% DPPH-methanol solution. A1 recorded at 517 nm af-
ter 30 min reaction at RT and 5 mL of DPPH solution were added as
a control to measure A0 using UV 1700 Spectrophotometer, Shimadzu,
Japan. The clearance rate (I%) of DPPH was estimated using the follow-
ing equation,
[( ) ]
I% = 𝐴0 − 𝐴1 ∕𝐴0 × 100
Where, A1 : Absorbance of the pectin samples, A0 : Absorbance of control
against methanol
2.3.5. Antioxidant activity by Ferric Reducing Antioxidant Power (FRAP)
Assay
Reducing power of jackfruit slime layer sample was determined
by FRAP assay with some modifications as described by Jin and
Yang, (2019). All the required solutions were freshly prepared before
use. FRAP reagent was made fresh by adding 300 mM acetate buffer of
pH 3.6 adjusted by using NaOH and HCl, 10 mM 2,4,6-tri(2-pyridyl)-
s-trizine in 40 mM HCl and 20 mM FeCl3 in the 10:1:1 ratio. 3 mL of
FRAP reagent were added into the 1 mL of each sample (mg/ml) and
kept at 37 °C for 30 min and reading was recorded at 593 nm (UV 1700
spectrophotometer, Shimadzu, Japan.).
M. Kumar, J. Potkule, M. Tomar et al. Carbohydrate Polymer Technologies and Applications 2 (2021) 100054

Fig. 1. Flow diagram of pectin extraction using acid extraction.

3
M. Kumar, J. Potkule, M. Tomar et al. Carbohydrate Polymer Technologies and Applications 2 (2021) 100054

Fig. 2. (A) Total phenolic content (B) Antioxidant activity using DPPH, (C) FRAP, (D) CUPRAC of JSS powder, JSSP, commercial citrus pectin, commercial apple
pectin. (E) Denotes the Pearson’s correlation among the total phenolic content and antioxidant activities. Where, JSSP: jackfruit slimy sheath pectin; JSS: jackfruit
slimy sheath; GAE: gallic acid equivalents; I%: percent inhibition; DPPH: 2,2-diphenyl picryl hydrazyl; FRAP: ferric reducing antioxidant power; CUPRAC: cupric
reducing antioxidant capacity. Values are expressed as means of triplicate measurements: Means with different alphabets are significantly different at p < 0.05.

2.3.6. Antioxidant activity by CUPRAC
CUPRAC stands for ‘cupric reducing antioxidant capacity’. Antioxi-
dant activity was measured as per Apak et al., (2004) which measures
the copper (II) or cupric ion reducing ability of the samples. This is
a simple and widely applicable antioxidant capacity index for dietary
polyphenols, vitamins C and E.
2.3.7. Fourier-transform infrared (FTIR) spectroscope analysis
FTIR spectroscope was determined by using IR Prestige 21, Shi-
madzu, Japan. A proper amount of sample was ground together with
the dried Potassium bromide (KBr) powder to form a pellet. After mak-
ing fine powder and pressing, the infrared absorption reading of sample
was taken at 400–4000 cm−1 . KBr was used as a control.
2.3.8. Scanning Electron Microscopy (SEM)
SEM imaging was performed using the method by Liew, Chin and
Yusof (2014) with some modifications. Surface microstructure of vari-
ous pectins was studied by SEM at 10 kV accelerating voltage and using
two magnifications 100× and 500×. 100× magnification was critical for
studying the uniformity particle size of pectins and 1000× was used to
study the surface characteristics of the individual pectin particle. The
samples were placed on the sample holder were vacuum sputtered with
gold-palladium mixture to improve its conductivity and viewed under
SEM.
2.4. Statistical analysis
The data reported in all the tables are an average of triplicate ob-
servation unless otherwise specified. Statistical significance of the vari-
ables viz., total phenolic content, antioxidant activity by DPPH, FRAP
and CUPRAC were evaluated through Kruskal Wallis test, a nonparamet-
ric one-way ANOVA for independent samples using IBM SPSS version
17, depicted in Fig. 2 as the assumptions of parametric ANOVA were
not met for these variables. The correlation between total phenolic con-
tent, antioxidant activity by DPPH, FRAP and CUPRAC was evaluated
by calculating their linear relationships through Pearson’s correlation
coefficient. The correlation study was performed using Jamovi version
1.2.27 at 5% level of significance.
3. Results and discussion
3.1. Extraction Yield of pectin
The organic acid (oxalic acid) was utilized for the pectin extraction
from the JSS. A maximum yield of 35.52% pectin was achieved from
JSS by using oxalic acid (0.05N) for an incubation time of 60 min at
90 °C. Sundarraj et al., (2018) reported coinciding results while extract-
ing pectin from the jackfruit peel waste using oxalic acid. Many other
researchers (Cho et al., 2019; Cui et al., 2020; Priyangini, Walde, &
Ramalingam, 2018), reported 16.24–21.28% of pectin from grapefruit
peel, 6.4% pectin from apple peel, 3.5–9.8% from pea hull pectin, 4.2–
74.5% from cocoa pod husks by using acid extraction methods. Xu et al.
(2018) reported 21.5% of jackfruit peel pectin extraction by microwave-
assisted extraction method. The higher yield of JSSP in case of oxalic
acid was attributed to the partial hydrolysis of the protopectin present
in cell wall into the smaller pectins during the extraction process (Chan
& Choo, 2013). Extraction yields of pectin (20.1%) from dragon fruit
peel using ammonium sulphate and oxalic acid was found highest which
is in agreement with our results (Muhammad et al., 2014). The higher
yield of JSSP was also contributed by the higher temperature (90 °C) and

4
M. Kumar, J. Potkule, M. Tomar et al.
Fig. 3. FTIR Spectra (a) jackfruit seed slimy sheath pectin (b) jackfruit seed slimy sheath powder (c) commercial citrus pectin (d) commercial apple pectin.
optimum time (1 h) of incubation for the hydrolysis of high polymeric
pectin into low polymeric pectin (Vriesmann et al., 2012).
3.2. Galacturonic acid (GalA)
The galacturonic acid content of JSSP, JSS powder, commercial ap-
ple and citrus pectin was determined by the spectrophotometric method
(Jin & Yang, 2019) using D-galacturonic acid as standard. Pectin con-
sist of 𝛼-1,4 linked galacturonic acid and includes rhamnogalacturo-
nan (RG) I, rhamnogalacturonan (RG) II, xylogalacturonan, and homo-
galacturonan. Pectin use as a food additive if they contain a minimum
of 65% galacturonic acid content (Flutto, 2003). GalA content of JSSP
(68.8 ± 2.57%) was higher than the JSS (28.8 ± 3.92) but less than
commercially available citrus (76 ± 3.32%) and apple (80.1 ± 3.98%)
pectin.
5
Carbohydrate Polymer Technologies and Applications 2 (2021) 100054
3.3. Total Phenolic Content (TPC)
TPC was measured by the Folin-Ciocalteu reagent and sodium car-
bonate method (Asgari et al., 2019) using gallic acid as a standard. It was
important to analyze the TPC of the extracted pectin as acid extraction
or hot water extraction leads to co-extraction of the phenolic compounds
(Sucheta, Misra, & Yadav, 2019). As evident from [Fig. 2 (A)], TPC
varied significantly (p < 0.05), TPC of the JSSP (65.7 mg GAE/g) was
found maximum compared to JSS powder (32.9 mg GAE/g), commer-
cial citrus (30.7 mg GAE/g) and apple pectins (51.3 mg GAE/g).The co-
extraction of phenolic compounds of JSS with pectin may be the reason
for the high phenolic content of JSSP. Parallel results were reported by
Sucheta et al. (2019) hot water extraction of black carrot pomace pectin
resulted in higher antioxidant activity compared to ultrasound-assisted
and microwave-assisted extraction of pectin because of co-extraction of
phenolic compounds and anthocyanins with pectin.
M. Kumar, J. Potkule, M. Tomar et al.
3.4. Antioxidant activity (AOA) by DPPH
DPPH is a well-established stable free radical when dissolved in al-
cohol it exhibits a peculiar absorption at 517 nm. Antioxidants from
the samples act as H+ donor and function as free radicals’ scavengers.
This converts the dark purple color of DPPH assay solution to light yel-
low, leading to a reduction in absorbance value (Nisar et al., 2018;
Kazemi, Khodaiyan, & Hosseini, 2019). DPPH (%) inhibition varied sig-
nificantly (p < 0.05) among all the samples with JSSP exhibiting the
highest (25.29 ± 4.03) followed by JSS (12.35 ± 4.13) and commercially
available citrus (3.91 ± 1.43) and apple (6.78 ± 2.52) pectin as shown in
Fig. 2 (B). The high scavenging activity of the JSSP compared to others
may be due to the higher content of phenolic compounds as discussed
6
Carbohydrate Polymer Technologies and Applications 2 (2021) 100054
Fig. 4. SEM images (a) jackfruit seed slimy
sheath pectin (Left: 100×, Right: 1000×) (b)
jackfruit seed slimy sheath powder (Left: 100×,
Right: 1000×) (c) commercial citrus pectin
(Left: 100×, Right: 1000×) (d) commercial ap-
ple pectin (Left: 100×, Right: 1000×).
in Section 3.2. Secondly, the DPPH scavenging activity of pectin is also
affected by the amount of galacturonic acid and molecular weight. JSSP
may have high galacturonic acid content with low molecular weight
which have assisted in quenching more free radicals (Qin, Liu, Cheng,
& Wang, 2019).
3.5. Antioxidant activity (AOA) by FRAP
FRAP was measured as described method (Jin & Yang, 2019) using
ferrous sulphate as a standard. FRAP assay usually assess the antioxi-
dant potential of plant source. Ferric salt is an oxidant and an antioxi-
dant compound present in plant sources acts as a reducing agent. Ferric
complex/electron-transfer reactions in which Fe3+ reduce to the Fe2+ .
M. Kumar, J. Potkule, M. Tomar et al.
JSSP showed higher AOA 10.4 ± 0.51 µM similar to the DPPH assay.
Significant (p < 0.05) difference observed in the AOA of JSSP (10.4
µM), JSS (6.36 ± 0.75 µM) and commercial pectins from citrus (4.21 ±
0.6 µM) and apple (5.96 ± 0.19 µM) [Fig. 2 (C)]. A direct correlation
was observed in the AOA and TPC of the samples which validated the
co-extraction of phenolic compounds with pectin. Kazemi, Khodaiyan,
Labbafi, Saeid Hosseini and Hojjati, (2019) also reported similar co-
relation while extracting pectin from Pistachio green hull.
3.6. Antioxidant assay by CUPRAC
JSSP showed higher AOA 44.36 ± 2.6 µmol trolox (TR)/g followed by
JSS (24.71 ± 1.83 µmol TR/g), apple pectin (18.47 ± 0.84 µmol TR/g),
and citrus pectin (15.68 ± 0.71 µmol TR/g) [Fig. 2 (D)]. The AOA using
CUPRAC assay was also found significantly higher in case of JSSP as
that of DPPH, and FRAP assays.
3.7. Correlation analysis
The correlation coefficient was studied among TPC, antioxidant ac-
tivity by DPPH, FRAP and CUPRAC [Fig 2 (E)]. The results of Pearson’s
correlation test indicated that all these parameters were positively corre-
lated with each other (Pearson’s r > 0). FRAP showed significant corre-
lation with DPPH (r = 0.982, p < 0.05). CUPRAC was significantly corre-
lated with TPC (r = 0.960, p < 0.05) and FRAP (r = 0.968, p < 0.05). Non-
significant positive correlations were observed with p > 0.05 between
DPPH and TPC; FRAP and TPC; CUPRAC and DPPH. Similar correlation
trends were observed by Sethi et al. (2020), Zhang et al. (2017). The
strong correlation between TPC and CUPRAC could result from the ten-
dency of phenolic compounds to accept an electron, forming reasonably
stable phenoxyl radicals, consequently disrupting the chain of oxidation
reactions in cellular components (Landete, 2012).
3.7. Fourier-transform infrared (FTIR) spectroscope: analysis of extracted
and commercial pectins
FTIR is an important spectroscopy technique use in the primary
structural study of pectin (Zhang et al., 2019). The infrared analy-
sis of the 4 samples viz. JSSP (a), JSS powder (b), commercial citrus
pectin (c) and commercial apple pectin (d) can be seen in Fig. 3. A
sharp peak can be seen around 3408, 3385, 3431 and 3421 cm−1 re-
spectively for (a), (b), (c) and (d). These peaks strongly resemble the
stretching vibration of the hydroxyl functional group. C–H groups of
D-galacturonic acid consists of CH, CH2 , and CH3 type of groups. The
vibrations of these C–H groups were found at 2920, 2927, 2935 and
2935 cm−1 respectively for (a), (b), (c) and (d). The signals in the
range of 1200–1450 cm−1 were annotated to the C–O–H bending and
C–O stretching vibrations. The peaks from 950–1200 cm−1 are char-
acteristic fingerprint confirming the occurrence of carbohydrate moi-
ety in all the four samples and represent C–O–C (ether linkage) and
O–H (hydroxyl group) of pyranose ring (Xu et al., 2018). The results
of FTIR analysis of extracted pectin from JSS showed a very similar
spectrum when compared with standard pectin from apple and citrus.
Coinciding results were obtained in the infrared analysis of other pectin
sources viz. tomato processing waste, fig skin, sunflower heads, cus-
tard apple peel, citrus (Govindaraj et al.,2018; Gharibzahedi, Smith,
& Guo, 2019; Shivamathi et al., 2019; Sengar, Rawson, Muthiah, &
Kumar Kalakandan, 2019; Muthusamy, Manickam, Murugan, Chen-
drasekar, & Pugazhendhi, 2019; Zhang, Zhang, Liu, Ding, & Ye, 2015).
3.8. Scanning Electron Microscopy (SEM) Imaging
SEM analysis was carried out to see the detailed surface microstruc-
ture of the extracted pectin from jackfruit seed slimy layer and to com-
pare the morphological characteristics of the extracted pectin with that
7
Carbohydrate Polymer Technologies and Applications 2 (2021) 100054
of commercial pectin from apple and citrus. Fig. 4 shows the microstruc-
ture of the JSSP obtained by acid extraction method (a), JSS powder
obtained by oven drying at 60 °C (b), commercial citrus pectin (c), com-
mercial apple pectin (d). The SEM analysis reveals that the pectin ex-
tracted from jackfruit seed slimy sheath with oxalic acid resulted in a
smooth and flattened microstructure at 100× with irregularities in the
particle size. Similarly, jackfruit seed slimy sheath powder was found to
variable size particle at 100×. The variability seen in the particle size at
100× in all the four-pectin sample may due to difference the crushing
method as (a) and (b) was crushed using mortar and pestle in the lab
(coarser and variable in size) and (c) and (d) obtained from the industrial
process (finer and constant in size). Further, (a) showed wrinkled sur-
face at 1000× which was comparable with the microstructure of (c) and
(d). An interesting finding in SEM images of (b) showed the presence of
globular structures at both 100× and 1000× (showed with arrowhead in
Fig. 4 (b). These globular structures may be due to the presence of globu-
lar proteins in the jackfruit seed slimy sheath powder. The hypothesis of
presence of protein is also confirmed by determining the protein content
as shown in supplementary Fig. 1. The microstructure of (b) appeared
rougher, irregular and ruptured which may be due to the presence of
other sugar, proteins, fibres and macromolecules. A similar pattern of
SEM images was observed by Muñoz-Almagro, Valadez-Carmona, Men-
diola, Ibáñez, and Villamiel, (2019), Taghi Gharibzahedi, Smith, and
Guo, (2019) in cacao pod husk and fig skin pectins respectively.
4. Conclusion
The JSSP was first time extracted by the acidic extraction using oxalic
acid and resulted in the modest recovery of 35.52 %. Phenolic content
has shown a positive correlation with the antioxidant activities using
DPPH and FRAP assays. JSSP demonstrated to have the highest antiox-
idant activities compared to commercial pectin from apple and citrus.
Microstructure and functional group analysis of JSSP displayed great
resemblance with the commercial pectin samples. The findings of the
current study provide an important basis of development of JSSP antiox-
idant extracts for use in food, health products, medicine, and cosmetics.
Declaration of competing interest
None
Supplementary materials
Supplementary material associated with this article can be found, in
the online version, at doi:10.1016/j.carpta.2021.100054.
References
Akter, B., & Haque, M. A. (2018). Utilization of jackfruit (Artocarpus heterophyllus) seed’s
flour in food processing: a review. The Agriculturists, 16(02), 131–142. https://doi.org/
10.3329/agric.v16i02.40351.
(2012). Jackfruit improvement in the asia-pacific region – a status report (p. 182). Bangkok,
Thailand: Asia-Pacific Association of Agricultural Research Institutions.
Asgari, K., Labbafi, M., Khodaiyan, F., Kazemi, M., & Saeid Hosseini, S (2019). High-
methylated pectin from walnut processing wastes as a potential resource: Ultrasound
assisted extraction and physicochemical, structural and functional analysis. Interna-
tional Journal of Biological Macromolecules. https://doi.org/10.1016/j.ijbiomac.2019.
10.224.
Chan, S. Y., & Choo, W. S. (2013). Effect of extraction conditions on the yield and chemical
properties of pectin from cocoa husks. Food Chemistry, 141, 3752–3758.
Cho, E. H., Jung, H. T., Lee, B. H., Kim, H. S., Rhee, J. K., Yoo, S. H., et al. (2019). Green
process development for apple-peel pectin production by organic acid extraction. Car-
bohydrate Polymers. https://doi.org/10.1016/j.carbpol.2018.09.086.
Cui, J., Ren, W., zhao, C., Gao, W., Tian, G., Bao, Y., Zheng, J., et al. (2020). The structure–
property relationships of acid- and alkali-extracted grapefruit peel pectins. Carbohy-
drate Polymers, Article 115524. https://doi.org/10.1016/j.carbpol.2019.115524.
Flutto, L. (2003). Pectin | properties and determination. Encyclopedia of Food Sciences and
Nutrition, 4440–4449.
Gharibzahedi, S. M. T., Smith, B., & Guo, Y. (2019). Pectin extraction from common fig skin
by different methods: The physicochemical, rheological, functional, and structural
evaluations. International Journal of Biological Macromolecules, 136, 275–283. https:
//doi.org/10.1016/j.ijbiomac.2019.06.040.
M. Kumar, J. Potkule, M. Tomar et al.
Govindaraj, D., Rajan, M., Hatamleh, A. A., & Munusamy, M. A. (2018). From waste to
high-value product: Jackfruit peel derived pectin/apatite bionanocomposites for bone
healing applications. International Journal of Biological Macromolecules, 106, 293–301.
https://doi.org/10.1016/j.ijbiomac.2017.08.017.
Hendel, N., Larous, L., & Belbey, L. (2016). Antioxidant activity of rosemary (Rosmarinus
officinalis L.) and its in vitro inhibitory effect on Penicillium digitatum. International
Food Research Journal, 23(4), 1725–1732.
Ho, G. T. T., Ahmed, A., Zou, Y.-F., Aslaksen, T., Wangensteen, H., & Barsett, H (2015).
Structure–activity relationship of immunomodulating pectins from elderberries. Car-
bohydrate Polymers,, 125, 314–322.
Jancy, S., Shruthy, R., & Preetha, R. (2019). Fabrication of packaging film reinforced
with cellulose nanoparticles synthesized from jack fruit non-edible part using response
surface methodology. International Journal of Biological Macromolecules. https://doi.
org/10.1016/j.ijbiomac.2019.09.066.
Jin, Y., & Yang, N. (2019). Array-induced voltages assisted extraction of pectin from grape-
fruit peel and its characterization. International Journal of Biological Macromolecules.
https://doi.org/10.1016/j.ijbiomac.2019.10.2.
Kazemi, M., Khodaiyan, F., & Hosseini, S. S. (2019). Eggplant peel as a high potential source
of high methylated pectin: Ultrasonic extraction optimization and characterization. LWT.
https://doi.org/10.1016/j.lwt.2019.01.060.
Kazemi, M., Khodaiyan, F., Labbafi, M., Saeid Hosseini, S., & Hojjati, M. (2019). Pistachio
green hull pectin: Optimization of microwave-assisted extraction and evaluation of its
physicochemical, structural and functional properties. Food Chemistry, 271, 663–672.
https://doi.org/10.1016/j.foodchem.2018.07.212.
Kruger, N. J. (2009). The Bradford method for protein quantitation. In The protein protocols
handbook (pp. 17–24). Totowa, NJ: Humana Press.
Kumar, M., Tomar, M., Saurabh, V., Mahajan, T., Punia, S., del Mar Contreras, M., &
Kennedy, J. F. (2020). Emerging trends in pectin extraction and its anti-microbial func-
tionalization using natural bioactives for application in food packaging. Trends in Food
Science & Technology.
Landete, J. M. (2012). Updated knowledge about polyphenols: Functions, bioavailabil-
ity, metabolism, and health. Critical Reviews in Food Science and Nutrition, 52(10),
936–948.
Liew, S. Q., Chin, N. L., & Yusof, Y. A. (2014). Extraction and characterization of pectin
from passion fruit peels. Agriculture and Agricultural Science Procedia, 2, 231–236.
https://doi.org/10.1016/j.aaspro.2014.11.033.
Liu, L., Cao, J., Huang, J., Cai, Y., & Yao, J. (2010). Extraction of pectins with different
degrees of esterification from mulberry branches to bark. Bioresource Technology, 101,
3268–3273.
Muhammad, K., Zahari, N. I. M, Gannasin, S. P., Adzahan, N. M., & Bakar, J. (2014). High
methoxyl pectin from dragon fruit (Hylocereus polyrhizus) peel. Food Hydrocolloids,
42, 289–297.
Muñoz-Almagro, N., Valadez-Carmona, L., Mendiola, J. A., Ibáñez, E., & Vil-
lamiel, M. (2019). Structural characterization of pectin obtained from cacao pod husk.
Comparison of conventional and subcritical water extraction. Carbohydrate Polymers.
https://doi.org/10.1016/j.carbpol.2019.04.040.
Muthusamy, S., Manickam, L. P., Murugan, V., Chendrasekar, M., &
Pugazhendhi, A. (2019). Pectin extraction from Helianthus annuus (sunflower) heads
using RSM and ANN modelling by a genetic algorithm approach. International Journal
of Biological Macromolecules. https://doi.org/10.1016/j.ijbiomac.2018.11.036.
Nayak, A. K., Pal, D., & Santra, K. (2015). Screening of polysaccharides from tamarind,
fenugreek and jackfruit seeds as pharmaceutical excipients. International Journal of
Biological Macromolecules, 79, 756–760. https://doi.org/10.1016/j.ijbiomac.2015.05.
018.
Nisar, T., Wang, Z.-C., Yang, X., Tian, Y., Iqbal, M., Guo, Y., et al. (2018). Characterization
of citrus pectin films integrated with clove bud essential oil: Physical, thermal, bar-
rier, antioxidant and antibacterial properties. International Journal of Biological Macro-
molecules, 106, 670–680. https://doi.org/10.1016/j.ijbiomac.2017.08.068.
Priyangini, F., Walde, S. G., & Ramalingam, C. (2018). Extraction optimization of pectin
from cocoa pod husks (Theobroma cacao L.) with ascorbic acid using response sur-
View publication stats
Carbohydrate Polymer Technologies and Applications 2 (2021) 100054
face methodology. Carbohydrate Polymers. https://doi.org/10.1016/j.carbpol.2018.
08.103.
Qin, Z., Liu, H.-M., Cheng, X.-C., & Wang, X.-D. (2019). Effect of drying pretreatment
methods on structure and properties of pectins extracted from Chinese quince fruit.
International Journal of Biological Macromolecules. https://doi.org/10.1016/j.ijbiomac.
2019.06.209.
Sengar, A. S., Rawson, A., Muthiah, M., & Kumar Kalakandan, S. (2019). Comparison of
different ultrasound assisted extraction techniques for pectin from tomato processing
waste. Ultrasonics Sonochemistry, Article 104812. https://doi.org/10.1016/j.ultsonch.
2019.104812.
Sethi, S., Joshi, A., Arora, B., Bhowmik, A., Sharma, R. R., & Kumar, P. (2020). Significance
of FRAP, DPPH, and CUPRAC assays for antioxidant activity determination in apple
fruit extracts. European Food Research and Technology, 246(3), 591–598.
Shivamathi, C. S., Moorthy, I. G., Kumar, R. V., Soosai, M. R., Maran, J. P., Kumar, R. S.,
Varalakshmi, P. et al., (2019). Optimization of ultrasound assisted extraction of pectin
from custard apple peel: Potential and new source. Carbohydrate Polymers, 115240.
doi:10.1016/j.carbpol.2019.115240
Sucheta, Misra, N. N., & Yadav, S. K (2019). Extraction of pectin from black carrot po-
mace using intermittent microwave, ultrasound and conventional heating: Kinetics,
characterization and process economics. Food Hydrocolloids, Article 105592. https:
//doi.org/10.1016/j.foodhyd.2019.105592.
Sundarraj, A. A., Thottiam Vasudevan, R., & Sriramulu, G. (2018). Optimized extraction
and characterization of pectin from jackfruit (Artocarpus integer) wastes using re-
sponse surface methodology. International Journal of Biological Macromolecules, 106,
698–703. https://doi.org/10.1016/j.ijbiomac.2017.08.065.
Suryadevara, V., Lankapalli, S. R., Danda, L. H., Pendyala, V., & Katta, V. (2017). Studies
on jackfruit seed starch as a novel natural superdisintegrant for the design and evalua-
tion of irbesartan fast dissolving tablets. Integrative Medicine Research, 6(3), 280–291.
https://doi.org/10.1016/j.imr.2017.04.001.
Taghi Gharibzahedi, S. M., Smith, B., & Guo, Y. (2019). Ultrasound-microwave assisted
extraction of pectin from fig (Ficus carica L.) skin: Optimization, characterization and
bioactivity. Carbohydrate Polymers, 114992. doi:10.1016/j.carbpol.2019.114992
Tramontin, D. P., Cadena-Carrera, S. E., Cruz, A. B., Bella Cruz, C. C., Bolzan, A.,
Quadri, M. B., et al. (2019). Biological activity and chemical profile of Brazilian jack-
fruit seed extracts obtained by supercritical CO2 and low pressure techniques. The
Journal of Supercritical Fluids, Article 104551. https://doi.org/10.1016/j.supflu.2019.
104551.
Vriesmann, L. C., Teofilo, R. F., & Petkowicz, C. L. de. O. (2012). In Extraction and char-
acterization of pectin from Cacao Pod Husks (Theobroma cacao L.) with citric acid: 49
(pp. 108–116). LWT - Food Science and Technology.
Xu, S. Y., Liu, J. P., Huang, X., Du, L. P., Shi, F. L., Dong, R., Cheong, K. L., et al. (2018).
Ultrasonic-microwave assisted extraction, characterization and biological activity of
pectin from jackfruit peel. LWT, 90, 577–582. https://doi.org/10.1016/j.lwt.2018.01.
007.
Zhang, B., Deng, Z., Tang, Y., Chen, P. X., Liu, R., Ramdath, D. D., & Tsao, R. (2017).
Bioaccessibility, in vitro antioxidant and anti-inflammatory activities of phenolics in
cooked green lentil (Lens culinaris). Journal of Functional Foods, 32, 248–255.
Zhang, L., Zhang, X., Liu, D., Ding, T., & Ye, X. (2015). Effect of degradation methods on
the structural properties of citrus pectin. LWT - Food Science and Technology, 61(2),
630–637. https://doi.org/10.1016/j.lwt.2014.11.002.
Zhang, Y., Hu, M., Zhu, K., Wu, G., & Tan, L. (2018). Functional properties and utiliza-
tion of Artocarpus heterophyllus Lam seed starch from new species in China. Interna-
tional Journal of Biological Macromolecules, 107, 1395–1405. https://doi.org/10.1016/
j.ijbiomac.2017.10.001.
Zhang, Y., Zhang, Y., Li, B., Wang, X., Xu, F., Zhu, K., & Chu, Z. (2019). In vitro hy-
drolysis and estimated glycemic index of jackfruit seed starch prepared by improved
extrusion cooking technology. International Journal of Biological Macromolecules. https:
//doi.org/10.1016/j.ijbiomac.2018.10.075.