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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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Corresponding author:
Abdalbasit Adam Mariod, Faculty of Sciences and Arts-Alkamil,
King Abdulaziz University, Jeddah, Saudi Arabia.
Email: basitmariod@yahoo.com
Yield%
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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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
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.
Lsd0.05
SE
OH concentration
in NaOH
0.1
0.2
0.5
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.
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CG
3000
1649.02
1537.16
1454.23
1319.22
1230.50
50
2941.24
%T
1078.13
100
1500
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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).
386
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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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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).
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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
389
Taste
Texture
1.6c
2.4a
2.1b
0.1286a
0.0259
1.3c
1.8b
2.9a
0.3854a
0.0361
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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