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Article

Extraction and characterization of gelatin from


two edible Sudanese insects and its applications
in ice cream making
Abdalbasit Adam Mariod1,2 and Hadia Fadul2

Abstract
Three methods were used for extraction of gelatin from two insects, melon bug (Coridius viduatus) and
sorghum bug (Agonoscelis versicoloratus versicoloratus). Extraction of insect gelatin using hot water gave
higher yield reached up to 3.0%, followed by mild acid extraction which gave 1.5% and distilled water extraction which gave only 1.0%, respectively. The obtained gelatins were characterized by FTIR and the spectra of
insects gelatin seem to be similar when compared with commercial gelatin. Amide II bands of gelatins from
melon and sorghum bug appeared around at 15421537 cm1. Slight differences in the amino acid composition of gelatin extracted from the two insects were observed. Ice cream was made by using 0.5% insects
gelatin and compared with that made using 0.5% commercial gelatin as stabilizing agent. The properties of
the obtained ice cream produced using insects gelatin were significantly different when compared with that
made using commercial gelatin.

Keywords
Amino acid, melon bug, sorghum bug, gelatin, ice cream, stabilizing agent, FTIR
Date received: 26 December 2013; accepted: 29 May 2014

INTRODUCTION
Gelatin is dened as a product obtained by the partial
hydrolysis of collagen derived from the skin, white connective tissue, and bones of animals. Gelatin is a gelling
protein, which has widely been applied in the food and
pharmaceutical industries. Most of commercial gelatin
(95%) is made from the hide of pigs and cattle, and the
remaining part (5%) comes from the bones of porcine
and bovine (Cho et al., 2005). The amount of gelatin
used in the worldwide food industry is increasing annually. However, frequent occurrences of bovine spongiform encephalopathy (BSE) and foot/mouth diseases
have been problems for human health and thus by-products of mammalians are limited in utility of processing
in functional food, cosmetic, and pharmaceutical
products. Therefore, the study of gelatin from sh
Food Science and Technology International 21(5) 380391
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DOI: 10.1177/1082013214541137
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by-products, such as skin, (Alfaro et al., 2014) skin


and bone (Gudmundsson, 2002), and from insects
(Mariod et al., 2011b) has increased for the replacement
of mammalian resources.
The melon bug Coridius (C.) viduatus (Heteroptera:
Dinioridae; formerly Aspongopus viduatus) and
the sorghum bug Agonoscelis (A.) versicoloratus versicoloratus (Heteroptera: Pentatomidae; formerly
Agonoscelis pubescens) are commonly known in
Sudan as am bqh (Watermelon bug) and endt aldrh
(Sorghum bug), respectively. The two insects belong
to suborder Heteroptera, order Hemiptera. In some
areas of Sudan, the collected bugs were extracted and
the obtained oil was used for cooking and some
1
Faculty of Sciences and Arts-Alkamil, King Abdulaziz University,
Jeddah, Saudi Arabia
2
Department of Food Science & Technology, College of
Agricultural Studies, Sudan University of Science & Technology,
Khartoum North, Sudan

Corresponding author:
Abdalbasit Adam Mariod, Faculty of Sciences and Arts-Alkamil,
King Abdulaziz University, Jeddah, Saudi Arabia.
Email: basitmariod@yahoo.com

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Mariod and Fadul


medicinal uses, e.g. against camels dermatological
infections. Insects oil extracted from Coridius viduatus
and Agonoscelis versicoloratus versicoloratus are used as
famine food in Western parts of Sudan and has traditional medicinal uses (Mariod et al., 2004). Mustafa
et al. (2008) reported a high antibacterial activity of
melon bug crude and the phenolic compounds-free
oils by agar diusion assay against seven bacterial
isolates.
Gelatin from insects may provide an alternative
source that is acceptable as a Halal product. In
Sudan, many edible insects are consumed in several
parts of the country, e.g. desert locust, sorghum and
melon bugs (Mariod et al., 2004, 2006). The approximate analysis of C. viduatus and adult A. v. versicoloratus adults showed 8.3 and 7.6% moisture, 27.0 and
28.2% crude protein, 54.2 and 57.3% fat and 3.5 and
2.5% ash on a dry matter basis, respectively.
The two bugs proteins contained 16 known amino
acids, including all of the essential ones. Compared
with the amino acid prole recommended by FAO/
WHO, the bug protein was of medium quality due to
its medium content of essential amino acids (Mariod
et al., 2011a).
Gelatins extracted from melon bug and sorghum bug
were studied (Mariod et al., 2011b). The two insects
showed 27.0 and 28.2% crude protein, respectively.
Extraction of gelatin using hot water gave a high
yield followed by mild acid and distilled water extraction. SDS-PAGE pattern showed low molecular weight
chains, and the two gelatins contained protein with
molecular weight of 40 kDa as main component. The
dierential scanning calorimetry thermograms results
conrmed no dierence between extraction methods
concerning the extracted gelatin quality. FTIR spectra
of melon and sorghum bug gelatins were similar and
the absorption bands were situated in more than six
bands in melon bug gelatin and only six bands in sorghum bug gelatin. Amide II bands of gelatins from
both melon and sorghum bug appeared at around
1554 cm1, while Amide I bands (17341632 cm1)
appeared only in melon bug. Mariod et al. (2011b) studied the microstructures of the insect gelatin examined
with the scanning electron microscope and showed that
melon bug exhibited the nest gelatin network with
very small voids. Melon bug gelatin showed the ner
structure with smaller protein strands and voids than
sorghum bug gelatin.
During sh gelatin extraction, the acid process refers
to the extraction that is carried out in an acid medium
(Gilbert et al., 2002; Gomez-Guillen and Montero,
2001), and in some cases an acid pretreatment before
an extraction is applied. The alkaline process refers to a
pretreatment of sh skin with an alkaline solution, in
most cases followed by neutralization with an acid

solution, so the extraction may be carried out in an


alkaline, neutral, or acid medium (Jamilah and
Harvinder, 2002; Montero and Gomez-Guillen, 2000;
Zhou and Regenstein, 2004). Choo et al. (2010) dened
ice cream as a frozen and aerated dairy-based dessert
that usually associated with happiness, pleasure and
fun. And they emphasized that the consumption of
ice cream psychologically evokes an enjoyable state of
a person. Ice cream is a popular dairy product throughout the world, and the ice cream formula backbone are
milk fat, milk solids not fat, sweetener, stabilizer, and/
or emulsiers (e.g. gelatin), water, and air (Varnam and
Sutherland, 1994). Citrus ber as a single stabilizer had
a positive eect on the melting resistance of ice cream,
and when combined with an emulsier, it produced
desirable ice cream properties (Dervisoglu and Yazici,
2006). Pintor et al. (2013) reduced up to 25% of butyric
and vegetable fats with 3% of inulin on ice cream formulations. Those authors obtained ice cream with good
textural and sensory characteristics. The substitution of
fat with inulin increased the ice cream mix viscosity,
improved air incorporation, and produced ice cream
with soft and homogeneous textures.
The objective of this study was to obtain an edible
gelatin from these two insect species with good functional properties, to compare it with commercial gelatin, and to study the possibility of using the
obtained gelatin in ice cream manufacturing.

MATERIALS AND METHODS


Samples preparation
Coridius viduatus and Agonoscelis versicoloratus versicoloratus adult bugs were collected from Ghibaish, West
Kordofan state and Abu Haraz, Gezira state in Sudan,
respectively; the collected bugs were stored in a tight
polyethylene bag, killed by treatment with hot water
for a few minutes and then sun dried (Mariod et al.,
2004), then ground to obtain homogeneous samples,
before representative samples were taken for chemical
analysis.
Chemicals and reagents
All solvents (n-hexane, diethyl ether, acetic acid,
NaOH) and chemicals (KBr pellets and 6 N HCL)
used were of analytical grade.
Defatting of samples
The ground-dried adult insect samples (200 g) were
weighed and put in an extracting bottle, and the samples were treated with diethyl ether (samples:solvents
1 : 2). Then the samples were put in a shaker (IKA ,
Werke GmbH & Co., Germany) and shaken at 240 rpm
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Food Science and Technology International 21(5)


for 24 h. After extraction, the samples were left to settle,
and then ltered through Whitman NO. 1. The insect
cakes were dried by air at room temperature to 24 h,
and the ltrates (oil with diethyl ether) were taken to
separate the oil from solvent using under vacuum
evaporator Model (1KA-WERKE\GMBH & Co-KG
D-79219 Staufen Germany).
Gelatin extraction
Gelatin extraction using distilled water and alkali
pretreatment. Following the method of Zhou and
Regenstein (2005) repeated by Mariod et al. (2011b),
cleaned, powdered and dried insects (20 g) were pretreated with 120 g of NaOH (1 : 6 w/v) with varying
NaOH concentrations (0.1, 0.2, and 0.5 mol/L) at 2
4  C for 60 min. The pretreated insects were drained
and rinsed with tap water and were then ready for the
gelatin extraction. After the pretreatment, samples were
mixed with 60 g distilled water (1 : 3 w/w, total 80 g) and
extracted at 50  C for 180 min in an incubator (EN 400Ankara-Turkey). The obtained insect gelatin was
freeze-dried in the freeze dryer Model (Edwards
Modulyo, Crawley, West Sussex, UK).
Gelatin extraction using mild acid and distilled
water. The cleaned defatted powdered insects (20 g)
were treated with a mild acid (50 mM acetic acid) as a
swelling step for 3 h. The pre-treated insects were
drained and rinsed with tap water and were then ready
for the gelatin extraction. After the pretreatment, samples were mixed with 60 g distilled water and overnight
(1618 h) gelatin was extracted at a moderate temperature (45  C) in the incubator (EN 400-Ankara-Turkey).
The extracted gelatin was freeze-dried in a freeze dryer
(Edwards Modulyo) (Gomez-Guillen et al., 2002; Zhou
and Regenstein, 2005; Mariod et al., 2011b).
Gelatin extraction using hot water. Following the
method of Muyonga et al. (2004), the cleaned defatted
powdered insects (20 g) were treated with a mild acid
(50 mM acetic acid) or NaOH with varying OH concentrations (0.1, 0.2, and 0.5 mol/L).The pretreated samples
as mentioned earlier were transferred to a beaker
covered with warm (above 60  C) water and gelatin was
extracted in a water bath (LWB-111D SCOTT
SCIENCE UK) by three sequential 5-h extractions at
50, 60 and 70  C followed by boiling for 5 h. Extraction
of pH was between 3.5 and 4. The obtained insect gelatin
was ltered through Whitman lter paper and freezedried in Freeze dryer Model (Edwards Modulyo).

Yield%

Weight of insect gelatin


 100
Weight of defatted insects powdered

Characterization of gelatin
Fourier transform infrared spectroscopy of
gelatin. The IR spectra were recorded with PerkinElmer Spectrum, model FTIR -8400s-Shimadzu
(Japan) spectrophotometer. Infrared data management
system (IRDM) was used during Fourier transform
infrared spectroscopy (FTIR) data collection. The
instrument was purged with dry nitrogen to remove
the moisture content, thus protecting the samples
from interference by water vapor and other impurities,
such as CO2. The prepared insect and commercial gelatin (obtained from various animal by-products) were
placed on a 13 mm magnetic lm holder and placed in
the spectrometer. The scanning of the background and
the gelatins were done using the software. The wave
number was set from 4000 cm1 to 400 cm1. The spectra of both background and gelatins were obtained
from 40 scans with the resolution of 2.0 and interval
of 0.5. Windows-based software program was used to
obtain the frequency of each band using the label
peaks command of the software or using the vertical
cursor by moving it to nd the frequency at the maximum absorbance for the selected band. The results
obtained were then collected for subsequent analysis
(Muyonga et al., 2004).
Amino acid composition by amino acid analyzer. The
content of dry matter and total N were determined
according to procedures described by AOAC (1990).
The content of amino acids (except for tryptophan) in
powdered gelatin (commercial, melon and sorghum bug
gelatin) was determined using Amino Acid Analyzer
(Sykam amino acid analyzer S433- Germany) under
the experimental conditions recommended for protein
hydrolyzates. Samples were acid hydrolyzed with
5.0 mL of 6 N HCl in vacuum-sealed hydrolysis vials
at 110  C for 24 h. The ninhydrine was added to the
HCl as an internal standard. Hydrolyzates were suitable for analysis of all amino acids. The tubes were
cooled after hydrolysis, opened, and ltered, and
then 200 mL of ltrate was taken and evaporated at
140  C for an hour and 1.0 mL of diluted buer was
added to the dried sample. The sample was ready
for analysis. Nitrogen in amino acids was determined
by multiplying the concentration of individual
amino acids by corresponding factors calculated from
the percentage N of each amino acid (Sosulski and
Imadon, 1990). The ammonia content was included
in the calculation of protein nitrogen retrieval, as it
comes from the degradation of some amino acids
during acid hydrolysis (Mosse, 1990; Yeoh and
Truong, 1996). The ammonia nitrogen content was calculated by multiplying the ammonia content by 0.824
(N 82.4% NH3).

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Manufacturing ice cream using insect gelatin. The ice
cream formula (ingredients) used in this study was as
follows: Fat 12%, MSNF (milk solids-not-fat; i.e. caseins, whey proteins and lactose) 10%, sugar (sucrose)
15%, gelatin 0.5%, egg yolk 0.3%, and water 62.2%.
Preparation of the ice cream mix. The mix (unfrozen
ice cream) was pasteurized using a double boiler to prevent scorching. The liquid ingredients (milk, cream)
were placed in the upper section of the double boiler,
and then whipped into the eggs and the skim-milk
powder. The gelatin was mixed with the sugar and
added to the liquid with constant mixing, stirred, and
heated to about 70  C. The container was placed in cold
water to cool as rapidly as possible to below 18  C
(Go and Hartel, 2013). The mixture was piped to
the pasteurization machine. The mix was homogenized
by the application of intensive air pressure (2000
pounds per square inch, 141 kg/cm2) and the hot mixture was forced through a small opening into the homogenizer. The ice cream mix was aged by storage in the
refrigerator overnight. This process improved the whipping qualities of the mix and the texture of the ice
cream. After the aging process was completed, the
mix was removed from the refrigerator and stirred in
the avoring (Go and Hartel, 2013). The mix was
frozen for 2030 min. The ice cream was scraped from
the dasher and scooped into suitable containers.
Immediately the ice cream was placed in a deep freezer
to harden to a temperature of 10 F (23  C). The ice
cream cartons were moved to a tunnel set at 30 F
(34  C). Constantly turning ceiling fans create a
wind chill of 60 F (51  C). The cartons moved
slowly back and forth through the tunnel for 23 h
until the contents are rock solid. The cartons were
then stored in the refrigerator (Fred, 1994). The developed ice cream was produced by adding 0.5% commercial gelatin (A), 0.5% insects gelatin (B), and 1.0%
insects gelatin (C) as stabilizing agents.
Sensory evaluation of the produced ice cream. Most
aspects of quality can be measured by sensory panels.
More tightly, food quality and acceptance can be indicated by correlating the quantitative and qualitative
measurements of the various sensory characteristics
contributing to the overall appreciation of the food,
with the consumer assessment or preference ratings
(Williams, 1970; Larmond, 1982). Sensory evaluation
has been dened as a scientic discipline used to
evoke, measure, analyze, and interpret those responses
to products as perceived through the senses of sight,
smell, touch, taste, and hearing. Ranking tests decide
on the attribute to be ranked, e.g. taste and texture, and
they allow tasters to evaluate samples and place them in
rank order according to the presence or absence of the

attribute from very to not at all and then record


the responses. In ranking tests, a small number (e.g. 8
15) of experienced panelists is used (Sidel and Stone,
1993). The produced ice cream (A, B, C) was subjected
to panel tests, taste and texture assessments were
obtained by using the ranking test, and the ranks
obtained by the panelists were converted to scores
and the results obtained by the panelists were statistically analyzed. Ten semi-trained panelists were selected
among the student of Department of Food Science and
Technology, Sudan University of Science and
Technology. The procedures for the samples preparation, the training of the panelists, and the establishment of the sensory attributes were based on Meilgaard
et al. (1999). The tests were conducted in the laboratory
of the Department of Food Science and Technology,
Sudan University of Science and Technology.
Statistical analysis. Each reported value is the mean of
determinations for triplicate samples prepared from
each extracted gelatin, and data generated were subjected to SAS software package Versions 9.3. Onefactor RCD was performed. Means were then tested
and separated using DMRT as reported by Steel et al.
(1997). When analysis of variance (ANOVA) revealed a
signicant (P  0.05) and highly signicant (P  0. 01)
eects, data means were compared using a least signicant dierence (LSD) test. To avoid zero standard deviation for amino acid composition, constant (plus/
minus) were added to the actual reading.

RESULTS AND DISCUSSION


Gelatin yield
Table 1 shows the yield of gelatin extracted from two
insects (sorghum bug and melon bug) using mild acid
and distilled water extraction. The melon bug provided
high yields than sorghum bug did. The yield of gelatin
extracted from sorghum and melon bugs using distilled
water with pretreatment of NaOH with varying concentrations 0.1, 0.2 and 0.5 mol/L was low compared with
that obtained from sorghum and melon bugs using mild
acid and distilled water extraction method shown in
Table 2. Sorghum bug obtained yield of 215, 250 and
287 mg/g is representing 1.07, 1.27 and 1.40%, respectively, while melon bug obtained yield of 176, 197,
218 mg/g, respectively, representing 0.88, 0.99 and
1.0%, respectively. The yields of gelatin extracted
from sorghum bug using distilled water seem to be
higher than that extracted from melon bug (Table 2).
Gelatin was extracted from sorghum bug using hot
water extraction pretreated with NaOH with varying
concentrations (0.1, 0.2 and 0.5 mol/L) shown in
Table 3. This high yield compared with the other two
methods (mild acid and distilled water and distilled
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Table 1. Yield of gelatin extracted from two edible insects using mild acid and distilled water extraction method
expressed as mg gelatin per g of powdered defatted insectsa
Weight (mg)

Yield (%)

Weight of SB
and MB

SBG

MBG

Lsd0.05

SE

SBG

MBG

Lsd0.05

SE

20 g

263.53  12.13b

293.43  3.39a

13.5291

4.0867

1.333  0.06b

1.463  0.06a

0.1285

0.0689

Mean  SD value (s) bearing same letter (s) within rows are not significantly different (P  0.05).
SB: Sorghum bug; MB: Melon bug; SBG: Sorghum bug gelatin; MBG: Melon bug gelatin.

Table 2. Yield of gelatin extracted from two edible insects using distilled water extraction method, pretreated with NaOH
(0.1, 0.2 and 0.5 g/L) expressed as mg gelatin per gram of powdered defatted insectsa
Weight of
SB and
MB (g)
20

OH concentration
in NaOH (mol/L)
0.1
0.2
0.5

Lsd0.05
SE 

Weight of SBG (mg)

Weight of MBG (mg)

Yield of SBG (%)

Yield of MBG (%)

215.30  0.35b
250.70  0.35ab
287.90  0.35a
69.09**
19.97

176.80  0.35b
197.60  0.35ab
218.50  0.35a
33.88**
9.791

1.077  0.35a
1.273  0.35a
1.400  0.35a
0.3791n.s
0.1095

0.884  0.35b
0.993  0.35ab
1.100  0.35a
0.1787*
0.05164

a
Mean value(s) bearing same letter(s) within columns (for each quality attribute) are not significantly different (P  0.05).
SB: Sorghum bug; MB: Melon bug; SBG: Sorghum bug gelatin; MBG: Melon bug gelatin.

Table 3. Yield of gelatin expressed as mg gelatin per g of powdered defatted insects


extracted from sorghum bug using hot water extraction method, pretreated with NaOH
(0.1, 0.2 and 0.5 g/L)a
Weight of SB (g)
20

Lsd0.05
SE 

OH concentration
in NaOH
0.1
0.2
0.5

Weight of SBG (mg)

Yield of SBG (%)

720.30  0.35a
530.30  0.50b
323.70  1.85c
1.798
0.5619

3.597  0.00a
2.650  0.01b
1.600  0.00c
0.00051
0.0001581

Mean  SD value(s) bearing same letter(s) within columns (for each quality attribute) are not differ significantly (P  0.05).
SB: Sorghum bug; SBG: Sorghum bug gelatin.

water extraction), the yield of gelatin using hot water


with 0.1 NaOH gave high yields of 720 mg/g representing 3.59%, followed by hot water with NaOH 0.2 mol/
L of 530 mg/g representing 2.65% and hot water with
NaOH 0.5 mol/L of 323 mg/g representing 1.6%. In the
low concentration of alkali NaOH, the yield was high
and vise to visor. Mariod et al. (2011) reported that
during insect gelatin extraction, alkaline and acid pretreatment showed the eects of removing non-collagenous proteins with minimum collagen loss, and from
our results it is clear that alkaline pretreatment followed by hot water extraction showed a better eect
than acid pretreatment and even better than alkaline

pretreatment followed by distilled water extraction.


These ndings were in good agreement with that of
Mariod et al. (2011) and Zhou and Regenstein (2005),
who reported that the alkaline and acid pretreatment
were critical for the yield and gel property of insect and
pollock skin gelatin, respectively.
Generally, extraction of gelatin from sorghum bug
using hot water extraction pretreated by varying concentrations (0.1, 0.2, and 0.5 mol/L) of NaOH gave
high yield and that is highly signicant dier
(p  0.05) in comparison with other extraction methods.
The yield signicantly (p  0.05) decreased when NaOH
concentration increased (Table 3). There was a

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Mariod and Fadul


signicant dierence in the gelatins obtained from the
two insects using mild acid extraction (Table 1), but the
highly signicant dierence (p  0.05) was found in
the amount of gelatin extracted from two insects
using the distilled water extraction method (Table 2).
FTIR of insect gelatin
The FTIR analysis was used to determine the types of
functional groups present in insects gelatin molecules.
The functional groups are represented by the peaks
obtained (Figures 15) and since the absorption is
based on the vibration mode of atoms and is very specic, each peak at dierent wave number will only
assign to specic functional groups (Bell, 1974). The
analysis was done in mid-infrared (mid-IR) region,
where the wave number ranges between 4000 cm1 to
400 cm1. Most IR applications employ the mid-IR.
Comparisons can be made between melon and sorghum
bug gelatin proles using dierent methods of extraction and with commercial gelatin through the assignment of specic functional compound in the specic
wavelength. Both gelatins (melon and sorghum bug gelatin) showed similar spectra, in which absorption bands
were situated at 610 bands.
The FTIR spectra of melon and sorghum bug gelatins are depicted in Figures 15.
Amide I band of gelatins extracted from melon and
sorghum bug at around 16501643 cm1 is compared
with commercial gelatin which is at 1649 cm1. Amide
II bands to both melon and sorghum bug gelatins at a
round of 15421533 cm1 and commercial gelatin at
1537 cm1 (Figures 15). Amide I and amide II bands
of gelatins are reported to be at 17001600 cm1 and
15601500 cm1 (Muyonga et al., 2004). Amide I band
of gelatins extracted from melon bugs using two

CG

3000

1649.02
1537.16
1454.23
1319.22
1230.50

50

2941.24

%T

1078.13

100

dierent methods of mild acid and distilled water


appeared at around 1652 cm1, to both gelatins used
distilled water extraction used pretreatment with
NaOH with three concentrations (0.1, 0.2 and
0.5 mol/L) and 1650 to gelatin used a mild acid
(Figure 2), amide II bands of gelatin from melon bug
used the two methods appeared at around (1542 and
1541 cm1) in MBG1 and MBG2, respectively.
It can be seen from Figure 2 that the FTIR spectra
exhibited by melon bug gelatin seem to be similar to
commercial gelatin (control), in the alkyl group-general
(29232921 cm1) both gelatin from melon bug at
2921 cm1, alkyl group-hydroxy or possibly amino substituent (about 14611411 cm1), aliphatic alcohol with
carbonyl substitution (about 10971120 cm1) in
MBG3 (1097 cm1). Small amide III bands that
appeared at around 11101230 cm1 in the regions
were attributable to both controls MBG2 and MBG4
and not found in MBG1and MBG3 gelatin samples.
But also Amide I band of gelatins from sorghum bug
used three dierent methods appeared around 1668
1643 cm1 (Figures 3 and 4), amide II bands appeared
at (15411537 cm1) in SBG1(1537), SBG4 (1541) and
SBG5 (1533), also similarly commercial gelatin
appeared at (1537 cm1), and amid II bands as alkyl
group-general around (29292925 cm1) to both gelatin extracted used distilled water with three varying
NaOH concentrations and gelatin extracted using hot
water with 0.1 mol/L (SBG5), but not found in gelatin
extracted using a mild acid and hot water with NaOH
concentration 0.2 and 0.5 (SBG6 and SBG7) Figure 4.
In the aliphatic secondary amide II or benzoguanamide
(32783244 cm1) in SBG2 and SBG4 not found in
other sorghum bug gelatin. Alkyl group-hydroxy or
possibly amino substituent (about 14541407 cm1).
From Figures 25 the FTIR spectra exhibited by

1500

Figure 1. Fourier transform infrared spectra of commercial gelatin.

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melon and sorghum bugs gelatin seem to be similar,
and also similar to control (commercial gelatin).
Generally, gelatin extracted from melon bug using
two methods and pretreated with varying NaOH concentrations (0.1, 0.2 and 0.5 mol/L) seem to be very
similar to those extracted from sorghum bug that
used three methods and pretreated with dierent
NaOH concentrations (0.1, 0.2, and 0.5 mol/L), except
that there were some bands in 991, 879, 703, 661, 659,
621,and 619 cm1 in sorghum bug gelatins that were
not found in melon bug gelatin.
Amino acid composition
The essential amino acids cannot be manufactured in
human bodies, but can be obtained from food. Eight
amino acids are generally regarded as essential for
humans: phenylalanine, valine, threonine, tryptophan,
isoleucine, methionine, leucine, and lysine (Young,
1994). Additionally, tyrosine (or aromatic amino
acids), deciency in these amino acids may hinder the
healing recovery process (Mat et al., 1994). Mariod
et al. (2011a) reported 360.5 and 268.8 mg/g crude

protein as total amino acids of powdered adults of


C. viduatus and A. versicoloratus, respectively.
The total amount of the essential amino acids found
in C. viduatus and A. versicoloratus was higher than the
values recommended by FAO/WHO (1991) (Mariod
et al., 2011a).
The amino acid prole of gelatin extracted from two
insects using dierent methods analyzed by amino acid
analyzer is presented in Table 4. Signicant dierence
(P  0. 05) in the amount of total amino acids was
found. The total amount of amino acids of insect gelatin was lower than that of the commercial gelatin.
The total amount of amino acids of sorghum bug
gelatin was higher than that of water melon bug.
Commercial gelatin had higher amount of glycine
(123.87 mg/g N) than sorghum bug gelatin extracted
using mild acid (51.76 mg/g N) and gelatin from
melon bug using distilled water (51.47 mg/g N), and
gelatin from melon bug using mild acid (50.70 mg/g
N); the least amount of glycine was reported in gelatin
extracted from sorghum bug using distilled water and
hot water methods which are 48.49 and 48.24 mg/g,
respectively.

3000

925.77

1097.42
1110.92
1045.35

1411.80
1342.36
1415.65
1342.36

1139.85
1076.21

1650.95
1542.95
1461.94
1402.15
1652.88
1541.02
1456.16
1402.15

2956.67
2921.96
2852.52
2956.67
2921.96
2852.52

MBG4

1741.60
1654.81
1652.88
1568.02
1564.16

MBG3

2956.67
2921.96
2852.52

%T

2956.67
2923.88
2852.52

MBG2

3284.55

MBGI

1500

Figure 2. Fourier transform infrared spectra of gelatin from melon bug (MBG1,MBG2, MBG3, and MBG4) extracted using
mild acid and distilled water method pretreated by alkali (NaOH) of three concentrations (0.1, 0.2, and 0.5).

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Mariod and Fadul

3000

786.90
744.47
703.97
621.04
588.25
514.96
387.67

1116.71

1274.86

1407.94
1344.29
1280.65
1120.56

1149.50
1122.49
1269.07

1541.02
1454.23
1396.37

1118.64

1413.72
1450.37
1398.30

1668.31
1589.23
1774.39
1654.81
1643.24
1620.09
1656.74

2968.24
2929.67
3278.76

3739.72

SBG4

2925.81
2856.38

SBG3

3244.05

%T

3392.55

SBG2

991.34

SBGI

1500

Figure 3. Fourier transform infrared spectra of gelatin from Sorghum bug (SBG1, SBG2, SBG3 and SBG4) extracted
using mild acid and distilled water method pretreated by alkali (NaOH) of three concentrations (0.1, 0.2, and 0.5 mol/L).

3000

1078.13

1230.50

1274.86

1116.71
1149.50
1122.49

1269.07
1118.64

1541.02
1454.23
1396.37

1450.37
1398.30

1413.72

1537.16
1454.23
1319.22

1649.02

1774.39
1654.81
1643.24
1620.09

2968.24
2929.67
3278.76

3739.72

SBG4

2925.81
2856.38

SBG3

3244.05

%T

3392.55

SBG2

1656.74

2941.24

CG

1500

Figure 4. Fourier transform infrared spectra of gelatin from Sorghum bug (SBG2, SBG3 and SBG4) extracted using hot
water method pretreated by alkali of three OH concentrations (0.1, 0.2 and 0.5 mol/L) compared to commercial gelatin
(CG).

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Food Science and Technology International 21(5)

3000

659.61

1108.99
1024.13
1091.63

1419.51
1344.29
1276.79

661.54
619.11

1078.13

1537.16
1533.30
1456.16 1454.23
1398.30
1319.22
1230.50

1649.02
1662.52
1566.09

SBG7

1417.58
1340.43

1660.60
1562.23

%T

879.48

SBG6

1647.10

2922.81
2854.45

SBG5

2941.24

CG

1500

Figure 5. Fourier transform infrared spectra of gelatin from Sorghum bug (SBG5, SBG6 and SBG7) extracted using hot
water method pretreated by alkali of three OH concentrations (0.1, 0.2, and 0.5 mol\L) compared with commercial gelatin
(CG).

Glycine is located at every third position of the triple


helix of collagen. Slightly lower glycine content was
observed in comparison with degradation induced by
the thermal process (hot water extraction) or by
endogenous proteases during gelatin preparation
(Muyonga et al., 2004).
Other protein bands might represent the contaminating proteins in gelatin, suggesting incomplete removal
of non-collagenous proteins prior to extraction. The
high signicant dierence was observed in the amount
of proline of both melon and sorghum bug in comparison with commercial gelatin, and also the commercial
gelatin had a higher amount of proline (200.74 mg/g N)
than gelatin extracted from sorghum bug using hot
water method (90.42 mg/g N), and gelatin extracted
from melon bug using mild acid method (70.01 mg/g
N), and gelatin from sorghum bug using mild acid
extraction (62.60 mg/g N), and gelatin from melon
bug using distilled water method (58.94 mg/g N), and
nally gelatin from sorghum bug using distilled water
method.
The amount of sulfur-containing amino acid
(methionine) in the commercial gelatin sample (control)
was high with amount of 5.18 mg/g N, when compared
with gelatin extracted from sorghum bug using hot
water method (5.11 mg/g N) followed by gelatin
extracted from melon bug using mild acid method
(4.60 mg/g N), and gelatin from sorghum bug using

distilled water extraction of 3.84 mg/g N, and gelatin


from melon bug using distilled water (3.22 mg/g N),
and the last was from sorghum bug using mild acid
method (2.91 mg/g N). Aromatic amino acid (tyrosine
and phenylalanine) levels were found in commercial
gelatin (control) as 1.34 and 15.87 mg/g N, respectively,
and these levels increased during hot water extraction
to 6.63 and 16.30 mg/g N, respectively. Tyrosine level in
gelatin of sorghum bug extracted using mild acid was
5.55 mg/g N, which was lower (4.16 mg/g N) than that
from melon bug using mild acid extraction, and gelatin
from sorghum bug using distilled water which was
3.58 mg/g N. The least is gelatin from melon bug
using distilled water, which was only 3.17 mg/g N.
Phenylalanine amount in gelatin from melon bug
using mild acid was 16.23 mg/g N which was higher
than that in gelatin from sorghum bug using mild
acid and also gelatin from melon bug using distilled
water of 11.77 and 11.50 mg/g N, respectively.

Ice cream developed using insects gelatin


Gelatin assists in absorbing some of the free water in
the ice cream mix and helps prevent the formation of
large crystals in the ice cream (Go and Hartel, 2013).
The used insect gelatin fullled all the legal requirements of the Sudanese Standards and Metrology

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23.50b  1.87
13.04a  1.19
8.23a  1.07
54.01b  4.65
123.87a  8.97
117.04a  7.45
38.52b  3.98
5.18a  2.54
13.47b  1.25
34.95a  3.92
1.34f  0.87
15.87c  2.09
23.76a  3.25
35.44a  2.64
18.05f  1.63
100.16a  6.66
200.74a  13.65
626.51

Aspartic acid
Thereonine
Serine
Glumatic acid
Glycine
Alanine
Valine
Methionine
Isoluecine
Leucine
Tyrosine
Phenylalanine
Histidine
Lysine
Ammonia
Arginine
Porlinie
Total

18.40c  1.53
8.37e  1.08
6.09c  1.04
43.52c  4.61
48.49e  4.67
18.11f  1.59
25.92f  3.85
3.84d  2.67
9.97f  0.97
20.19f  2.67
3.58d  1.17
10.93f  1.96
7.39f  0.51
21.01e  2.51
30.56c  2.54
35.12e  2.78
52.86f  4.67
311.55

SBG (D.W)
14.60d  1.46
9.45d  1.09
5.62d  1.02
35.70d  4.58
51.76b  4.61
21.42d  4.23
31.65d  3.76
2.91f  2.63
11.36d  1.12
23.69d  3.74
5.55b  2.47
11.77d  1.69
9.94c  0.98
24.94c  2.39
35.62a  2.69
51.03c  4.92
62.60d  5.98
347.09

SBG (AA)
27.60a  2.91
12.59b  1.17
7.95b  1.05
63.94a  4.73
48.24f  4.53
24.09b  4.27
43.27a  4.02
5.11b  3.52
14.78a  1.28
31.84b  3.89
6.63a  3.11
16.30a  1.74
14.33b  2.97
32.20b  2.55
31.20b  2.57
58.07b  2.95
90.42b  7.18
438.20

SBG (HW)
13.34f  1.22
7.58f  1.07
4.30f  0.95
27.10e  4.43
51.47c  4.58
21.08e  4.19
28.96e  3.89
3.22e  2.61
10.28e  1.01
20.63e  2.68
3.17e  1.13
11.50e  1.28
8.53e  0.93
18.37f  1.66
28.51d  2.48
33.87f  2.67
58.94e  6.03
291.99

MBG (D.W)
13.94e  1.33
9.99c  1.13
4.50e  0.96
24.88f  4.38
50.70d  4.67
23.48c  4.25
37.31c  3.91
4.60c  2.55
12.90c  1.23
25.57c  2.95
4.16c  1.25
16.23b  1.72
9.78d  0.96
23.29d  2.30
28.03e  2.51
40.95d  3.86
70.01c  6.52
330.39

MBG (AA)

18.5263*
21.5241*
13.3621*
32.5437*
17.8765**
21.5243**
13.2906*
18.5629*
28.9814*
23.0357*
14.8762*
5.8544*
15.9813*
17.8194*
19.1396*
23.8647**
41.5928**

Lsd0.05

9.7345
6.5873
3.2941
10.5682
3.3974
8.3120
2.5807
6.9014
8.4693
3.0155
1.6842
4.9744
3.8746
5.7498
3.8539
6.5247
12.6830

SE

Note: Mean  SD value(s) bearing same letter(s) within rows (for each amino acid) are not significantly different (P  0.05).
*Statistically significant at 0.05 level, **statistically significant at 0.01 level, ***statistically significant at 0.001 level.
SBG (DW): gelatin from sorghum bug used distilled water extraction; SBG (MA): gelatin extracted from sorghum bug using mild acid method; SBG (HW): gelatin extracted from sorghum
bug using hot water method; MBG (DW): gelatin extracted from melon bug using distilled water method; MBG (MA): gelatin extracted from melon bug using mild acid method.

Control (CG)

Amino acid
(mg/100 g)

Samples

Table 4. Amino acid composition (mg/g) of gelatin extracted from two insects

Mariod and Fadul

389

Food Science and Technology International 21(5)


Table 5. Preference ranking of formulated ice cream
(B, C) as compared to commercial ice cream (A)
Sample

Taste

Texture

A: Commercial gelatin (0.5%)


B: SBG MBG (0.5%)
C: SBG MBG (1.0%)
Lsd0.05
SE 

1.6c
2.4a
2.1b
0.1286a
0.0259

1.3c
1.8b
2.9a
0.3854a
0.0361

The FTIR spectra exhibited by melon and sorghum


bugs gelatin seem to be similar, and also similar to the
commercial gelatin. Amino acid composition of insects
gelatin did not vary greatly, and even did not vary
depending on the method of extraction. The developed
ice cream products (B, C) showed a signicant
(P  0.05) dierence with regard to their taste and texture when compared with the commercial gelatin ice
cream (A).

Mean  SD value(s) bearing same letter(s) within columns (for


each quality attribute) are not significantly different (P  0.05).
SBG: Sorghum Bug Gelatin; MBG: Melon Bug Gelatin.

Organization (SSMO) for ice cream and was categorized as safe ice as adult insects are usually used in
Sudanese diet.
Sensory analysis of developed ice cream using ranking test. The results of analysis of variance (ANOVA)
of the ranks obtained by the panelists for ice cream,
processed using insect gelatin (B and C) in comparison
with that produced using commercial gelatin (A) as
control are presented in Table 5. It is clear that the
dierence was signicant (P  0.05) between the developed products of ice cream as regards to its taste and
texture. The ice cream produced using commercial gelatin (A) gave better taste and melt mouse than that with
insect gelatin (B and C). The results of analysis of variance (ANOVA) of the ranks for the taste and texture of
the products obtained by the panelists are presented in
Table 5. It is clear that the dierence between the three
developed ice cream products (A, B, and C) was signicant (P  0.05) as regard to their taste and texture,
which means the commercial gelatin ice cream (A)
has a better taste and texture than that of insect gelatin
ice cream (B and C), and ice cream B is better than C.

CONCLUSIONS
The gelatin extracted from the two insects C. viduatus
and A. versicoloratus using three dierent methods
showed good results in the yield. Extraction of gelatin
using hot water gave a high yield and reached up to
3.0%, followed by mild acid extraction of 1.5% and
distilled water extraction of 1.0%. During insect gelatin
extraction, alkaline and acid pretreatment showed the
eects of removing non-collagenous proteins with minimum collagen loss, and alkaline pretreatment followed
by hot water extraction showed a better eect than
acid pretreatment and even better than alkaline pretreatment followed by distilled water extraction.
Extraction of gelatin by hot water pre-treated with
varying concentrations of NaOH was found as the
best method for extraction of gelatin from insects.

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