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

net/publication/314199407

The effect of pretreatment using hydrochloric acid on the characteristics of

buffalo hide gelatin

Article · February 2017

DOI: 10.14710/jitaa.42.1.14-22

CITATION READS

1 335

4 authors, including:

Sri mulyani Mulyani Yudi Pranoto

Universitas Diponegoro Universitas Gadjah Mada
15 PUBLICATIONS   13 CITATIONS    57 PUBLICATIONS   900 CITATIONS   

SEE PROFILE SEE PROFILE

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

Nanoencapsulation of liquid smoke and bioactive compounds and its application as natural food preservative View project

All content following this page was uploaded by Yudi Pranoto on 13 March 2017.

The user has requested enhancement of the downloaded file.

 J. Indonesian Trop. Anim. Agric.
J I TAA pISSN 2087-8273 eISSN 2460-6278
http://ejournal.undip.ac.id/index.php/jitaa
42(1):14-22, March 2017
Journal of the Indonesian Tropical Animal Agriculture
Accredited by Ditjen Penguatan Risbang No. 60/E/KPT/2016 DOI: 10.14710/jitaa.42.1.14-22

The effect of pretreatment using hydrochloric acid on the

characteristics of buffalo hide gelatin
S. Mulyani1,3, F. M. C. S. Setyabudi2 , Y. Pranoto2 and U. Santoso2
1Doctoral Program, Faculty of Agricultural Technology, Gadjah Mada University,
Jl. Flora No.1, Bulaksumur, Yogyakarta 55281 - Indonesia
2Faculty of Agricultural Technology, Gadjah Mada University,
Jl. Flora No.1, Bulaksumur, Yogyakarta 55281 - Indonesia
3Permanent Address: Faculty of Animal and Agricultural Sciences, Diponegoro University,
Tembalang Campus, Semarang 50275 – Indonesia
Corresponding E-mail : umar_s@ugm.ac.id

Received August 31, 2016; Accepted January 04, 2017

ABSTRAK

Tujuan penelitian adalah untuk mengidentifikasi variasi konsentrasi asam klorida terhadap yield
dan sifat fisikokimia gelatin kulit kerbau. Design penelitian menggunakan Rancangan Acak Lengkap,
dengan perlakuan variasi konsentrasi asam klorida 0.3M, 0.6M, 0.9 M, 1.2 M dan 1.5M, ulangan
sebanyak empat kali, dilanjutkan uji beda nyata terkecil. Yield, kekuatan gel, viskositas, pH dan
intensitas warna merah gelatin dipengaruhi (P<0.05) oleh variasi konsentrasi HCl. Yield tertinggi
(29.17%) diperoleh pada konsentrasi 0.9M. Kekuatan gel dan viskositas tertinggi pada konsentrasi
1.2M, masing-masing 293.41 g bloom dan 22.17 cP. Gelatin kulit kerbau mempuyai pH 5.08 -5.52.
Tingkat kecerahan warna gelatin (L*) 69.92 – 70.97, sedangkan warna merah (a*) 0.55-1.54 dan kuning
(b*) 17.54 -19.59. Kadar air gelatin 7.05±0.07 – 8.92±0.06%, kadar protein 83.38±0.06 – 91.11±0.03%
dan kadar abu 0.53±0.04 – 1.23±0.09%. Berdasarkan absorbsi spectra fourier transform infrared (FTIR)
terdeteksi empat area amida yaitu amida A (3402.43 – 3448.72 cm-1), amida I (1635.65 cm-1), amida II
(1527.62 cm-1) dan amida III (1242.16 cm-1). Variasi konsentrasi HCl mempengaruhi yield, karakteristik
dan struktur sekunder gelatin kulit kerbau. Konsentrasi 0.9M merupakan kondisi optimum untuk
memperoleh yield tertinggi. Karakteristik gelatin kulit kerbau dengan pretreatment menggunakan asam
klorida telah memenuhi standar kualitas Gelatin Manufacturing Institute of America (2012) .
Kata kunci : asam klorida; gelatin , kulit kerbau, pretreatment, yield

ABSTRACT

The objective of the research was to identify the effect of the concentration of hydrochloric
acid (HCl) on yield and the characteristics of gelatin from buffalo hide. A completely
randomized design was used with various concentrations of HCl treatment 0.3M, 0.6M, 0.9 M,
1.2 M and 1.5M, and four times replication, followed by least significant difference test. The
HCl concentration gave significantly effect (P<0.05) on yield, viscosity, pH, gel strength and
redness of gelatin. The highest yield (29.17%) was obtained at a concentration of HCl 0.9M.
The highest gel strength and viscosity at concentration of 1.2M, were 293.41g bloom and
22.17 cp, respectively. The pH of the gelatin were 5.08 to 5.52. The lightness level (L*)
was 69.92-70.97, whereas the redness r a n g e d (a*) from 0.55 to 1.54 and the yellowness r a n g e d

14 J.Indonesian Trop.Anim.Agric. 42(1):14-22, March 2017

(b*) from 17.54 to 19.59. The moisture, protein and ash concentrations of gelatin were 7.05±0.07-
8.92±0.06%, protein content of 83.38±0.06 -91.11±0.03%, and the ash content 0.53 ± 0.04 to 1.23 ±
0.09%, respectively. Based on the spectra absorption of infrared were detected four areas amides: amide
A (3402.43-3448.72 cm-1), amide I (1635.65 cm-1), amide II (1527.62 cm-1) and amide III (1242.16
cm-1). The differences in the concentration of HCl had impact on yield, characteristics and secondary
structure of buffalo hide gelatin. The HCl concentration at 0.9M was the optimum condition to obtain
the highest yield. The characteristics of buffalo hide gelatin after pretreatment using hydrochloric acid
had met the standards of GMIA (2012) .
Keywords: Hydrochloric acid, gelatin, buffalo hide, pretreatment, yield

INTRODUCTION higher acid concentration may cause over-

hydrolysis of collagen molecules and losing
Buffalo is the one of tropical livestock protein during washing process (Binsi et al.,
producing skin by-product that may be used as 2009; Jamilah and Harvinder, 2002). The
raw material for producing gelatin. The skin by- hydrolysis rate of the collagen molecules
product of buffalo had thick and strong structure influences the yield and characteristics of gelatin.
because slaughtering age buffalo was older than Therefore, the optimization of the acid
slaughtering age of cows and commonly used as concentration prior to the extraction is required to
working livestock (Huda et al., 2011). The avoid over-hydrolysis. This study was aimed to
thickness of skin by-product is 6-8 mm with clarify the effect of the concentration of the HCl
approximately 11.5 % of the animal live weight concentration on the gelatin yield and the
(Spanghero et al., 2004). The thickness and characteristics of buffalo hide.
firmness of skin by-product indicated. A high
collagen fibers. A high collagen of the hide is MATERIALS AND METHODS
supported by the high pyrrolidine group of amino
acid hydroxyproline.. The pyrrolidine group The extraction of buffalo hide gelatin
attributes the collagen with a high thermal Wet salt preserved buffalo hide was obtained
stability (Haugh and Draget, 2009), so heat from CV. Panji Jaya in Cegoroyoso village of
extraction only could not optimize the extraction Pleret sub district of Bantul, Yogyakarta with the
process. Therefore, acid pretreatment process is criteria of male buffalo of 2-3 years. The hide
required to optimize the extraction process (Niu et washed using tap water and then the hide was
al., 2013). soaked in 2% (w/v) camphor solution till the hair
The acid pretreatment results in increasing was completely removed. Subsequently, the
weak intermolecular and intra- molecular processed hide was scratched to remove the hair
structural bond in the skin collagen protein and the fat and then was washed using tap water
because of partial breakdown of the amino acid till its pH was 7-7.5. After the removal of the hair
bond chain (Kolodziejska et al., 2007). This and the fat, the processed hide was kept frozen at
condition is used in the extraction process -18º C (Said et al., 2011). Before the pretreatment,
resulting in collagen solubility (Johnston-Banks, it was thawed at 4ºC for 20 hours and cut into
1990). Acid pretreatment at different acid small pieces (1x1cm) (Ktari et al., 2014).
concentrations has an impact on yield and gel The gelatin from buffalo hide was extracted
strength (Ahmed and Benjakul, 2011; Nikoo et according to the method of Niu et al. ( 2013)
al., 2014). An acid helps the release and dissolve with modification. The pieces of the buffalo hide
of some proteins, fat and other components of were soaked in 0.5M NaOH (1:4;w/v) for 2 hours.
collagen. This also disturbes cross-interaction Subsequently, the hide was soaked in HCl at the
molecules of the collagen thereby increasing the concentrations of 0.3M, 0.6M, 0.9M, 1.2M
efficiency of the extraction. Hydrochloric acid is and 1.5M (1:4;w/v). The processed hide was
a type of strong acid that could be dissociated drained and washed six times till reaching the pH
perpectly, thus producing many hydrogen ions to 5-6. The pretreated hide was extracted with
break crosslinks in collagen molecules. distillated water (1:4;w/v) at 65ºC in water bath
Hydrochloric acid requires lower concentration to (Memmert WNB7-45 type) for 5 hours and then
achieve an optimal extraction condition compared followed in similar manner at 70ºC for the second
to other type of acid (Niu et al., 2013).More step extraction. The extracting result was roughly

Buffalo Hide Gelatin by HCl Pretreatment (S. Mulyani et al.) 15

sieved and dried using cabinet drier at 50-55ºC for expressed in L, a, and b value was used to
48 hours. The extracted gelatin yield was measure the color of gelatin. The L value for the
calculated using formula of Ktari et al. (2014): lightness were in the range of 100 = white and 0 =
black. The a values were in the range of -50
Dry gelatin we ight
Yield (%)  x 100 ( green) and +50 (red). The b values were in the
Fresh hide weight range -50 (blue) and (yellow) (Jamilah and
Harvinder, 2002).
Proximate Analysis of Hide and Gelatin
The moisture, protein, and ash contents Determination of Fourier Transform Infrared
of the buffalo hide and gelatin were determined (FTIR) Spectroscopy
using proximate analysis following AOAC Gelatin powder (2 mg) was made pieces
guideline (2005). Analysis of each proximate was in 100 mg of kalium bromide (KBr). The sample
done in three replication. strip was read through FTIR (Shimadzu PC-8201)
in the range of wave number 4000-650 cm-1
Determination of Hydroxiproline (Kaewruang et al., 2013).
A hundred milligram samples of skin or
gelatin was crushed and hydrolyzed by adding Statistical Analysis
5ml 6N HCl for 12 hours at 110ºC in dry- This study was designed using complete
bath. Then, NaOH 6 N was used to neutralize the random design with various HCl concentration as
samples. Two ml acetate or citrate buffer and 0.3 treatments with four replicates. The Analysis of
M NaCl were added till 25 ml volume was variance and least significant difference test (Steel
reached. The solution was transferred to the and Torri, 1980) were performed using SPSS 20.0
300 µl isopropanol and oxidant solution for Windows. Especially for proximate,
containing testing flask. At this time the mixture hydroxyproline and FTIR were determined using
was quickly mixed for 4 minutes, and then was descriptive analysis based on the average mean
added 4 ml of Ehrlich's solution in 3 ml of 60% with three times replicates.
acid per chloride (v/v). The solution mixture was
shake for 25 minutes in water bath (60ºC). The RESULTS AND DISCUSSIONS
optical density of solution mixture was measured
using a spectrophotometer at wavelength of 660 Yield
nm. The calculation of hydroxyproline content Yield is often related to the extraction
was followed by standard hydrokxyproline efficiency in gelatin production. The increasing
solution curve (Sigma Chemical). high yield is indicative of production process
efficiency (Kolodziejska et al., 2007). The
Determination of Gel Strength gelatin extraction yield at various HCl
Gelatin powder (6.67 g) was dissolved in concentrations in the pretreatment process are
100 mL of distilled water (6.67%; w/v) and was shown in Table 1. The values were in the range of
stirred using magnetic stirrer and then heated at 14.67% to 29.17%, with the highest yield at the
60ºC for 15 min and incubated 16-18 hours at HCl concentration of 0.9 M. The values
10ºC. Subsequently, it was measured using texture decreased to 25.59% and 24.53%, when the HCl
analyzer TA-XT plus HD (Stable Micro System concentration was increased up to 1.2 and 1.5 M.
Ltd., UK) at the probe speed of 0.5 mm/sec and 4 The HCl concentrations gave significantly
mm depth (Benjakul, et al, 2009). affect (P<0.05) on the yield of buffalo hide
gelatin. Acid pretreatment is required as H+ ion
Determination of Acidity (pH) provider that plays an important role in breaking
Zero point two gram sample was weighted, both intra and intermolecular bonds of collagen,
dispersed into 20 ml of distilled water at 80ºC, so collagen is easily dissolve in water. The
and homogenized using magnetic stirrer. The increasing acid concentration was followed by the
acidity of solution was measured using pH meter increased H+ concentration in solution, finally the
(British Standard 757). collagen hydrolysis process was accelerated. The
higher hydrolysis rate caused the triple helix
Measurement of Color collagen breakdown into more bigger α, β and ϒ
Chromameter (Konika Minolta Sensing, chains. In other words the collagen conversion
INC, Japan) with the Hunter system and into gelatin increased in accordance with the

16 J.Indonesian Trop.Anim.Agric. 42(1):14-22, March 2017

Table 1. Yield and the Characteristics of Buffalo Hide Gelatin by Pretreatment Using HCl

Concentration Gel Strength Viscosity* Measurement of Color

Yield* (%) pH*
of HCL (g bloom)* (cP) L ns
a* bns
0.3 M 14.67±1.40c 265.08±3.64b 18.65±1.20ab 5.52±0.03a 69.92±1.94 0.55±0.22a 17.54±0.59
0.6 M 26.44±2.10b 239.44±13.45c 17.92±1.20b 5.16±0.02b 70.97±0.91 1.54±0.19b 18.90±0.58
0.9 M 29.17±2.10a 242.35±12.90c 16.37±1.50b 5.15±0.05bc 70.66±1.46 1.51±0.13b 18.14±1.52
1.2 M 25.59±0.80b 293.41± 7.30a 22.17±1.70a 5.09±0.04c 70.55±0.47 1.21±0.56b 19.59±1.80
1.5 M 24.53±0.90b 248.94±13.45bc 18.23±2.80ab 5.08±0.03c 70.82±0.83 1.27±0.98b 17.87±1.24
* Means within the same column followed by different superscript are significant different (P<0.05); ns = non-
significant (P>0.05). L: lightness, a: redness, b; yellowness

increasing yield value. On the contrary, the collagen structure became more open and weak.
increase in the HCl concentration more than 0.9M The water holding capacity lessened and
resulted in the decrease in the yield. Continuing increased the quantity of water evaporation
hydrolysis process may cause more shorter during drying process (Sompie et al. 2015). The
collagen molecule chain and hence there were moisture of buffalo hide gelatin ranged from
many dissolved collagen in the washing process. 7.05 ± 0.07 to 8.92 ± 0.06%. These value were
Once an optimum condition has been met, the lower than the results of Alfaro et al. (2014) and
increase in the acid concentration would decrease standard commercial gelatin is 9-14% (Eastoe and
the gelatin yield because of the over-hydrolysis of Leach, 1977).
the collagen and there were many proteins lost in The protein content of gelatin directly related
the washing process (Niu et al., 2013: Jamilah and to the gel strength and viscosity (Said et al.,
Harvinder, 2002). 2011). The protein content of buffalo hide gelatin
were in the range of 83.38 ± 0.06 to 91.11 ±
Chemical Composition of Hide and Gelatin 0.03%. ( T a b l e 2 ) . T h i s r e s u l t w a s
The composition of the hide and the gelatin almost consistent with other studies. Said et al.
from buffalo was represented in the Table 2. The (2011) examined the gelatin of goat skin, the
raw material of the wet salt preserved buffalo hide protein content were in the range of 89.37
has gone through the process of soaking, so that it ±1.19% - 90.74 ± 1.82%. Sompie et al (2015)
looked like fresh skin. Said et al. (2011) reported reported that gelatin from pig skin, were in the
that fresh skin contained 64% moisture, 33% range of 86.03 - 89.22%. This result was also
protein and 0.2% mineral. Alfaro et al. (2014) similar to commercial gelatin approximately at
suggested that the protein content in the hide 89.63% (Pranoto et al., 2006). The collagen
described maximum collagen content found in dissolving rate is influenced by the material and
skin tissue. time used for the pretreatment process (Wang
In general, the hydroxyproline content of the et al., 2008; Said et al., 2011). The protein
buffalo gelatin was lower than the hidroxyproline content of gelatin from buffalo hide increased
buffalo hide (% dry matter). The collagen from 0.3M to 0.6 M and 0.9 M. The increase in
conversion process into gelatin was carried out HCl concentration resulted more broken crosslink
through pretreatment and extraction process at 65- of both intra and intermolecular collagen that
70ºC so that the hydrolysis process caused the there are many proteins dissolved in the extraction
decrease in the hydroxyproline content. process. The big number of the dissolved protein
The moisture content of gelatin tended to caused the increase in the protein content of
diminish by the increasing HCl concentration gelatin. At the concentration of 1.2 M and 1.5 M
used in the pretreatment process. The acid the protein content did not further increase
treatment brokedown both the intra- and because the optimum concentration has been met.
intermolecular collagen crosslink so that the The increase in the acid concentration would

Buffalo Hide Gelatin by HCl Pretreatment (S. Mulyani et al.) 17

Table 2. Chemical Composition of Hide and Gelatin from Buffalo by Pretreatment Using HCl

Buffalo Hide Gelatin

Composition (%) Buffalo Hide
HCl 0.3M HCl 0.6M HCl 0.9M HCl 1.2M HCl 1.5M
Moisture 68.62±0.13 8.92±0.06 7.41±0.05 7.09±0.01 7.05±0.08 7.64±0.21
Protein 30.22±0.06 89.35±0.01 83.38±0.06 91.11±0.03 90.93±0.01 90.89±0.03
Ash 0.72±0.01 1.23±0.09 0.87±0.07 0.56±0.01 0.59±0.04 0.53±0.04
Hydroxy-prolin 0.25±0.04 2.18±0.02 0.71±0.02 0.44±0.01 0.46±0.01 0.46±0.02
Results are means ± standard deviation (n=3)

result in over-hydrolysis, therefore many proteins improved the effect in formation process of gel.
lost during washing (Niu et al., 2013; Sompie et Subsequently, amino acid monomer chains joined
al., 2015). to form a continuous three-dimensional structures
The gelatin extract from buffalo hide had the and bond water to form compact gel structure.
ash content approximately from 0.53 ± 0.04 to The difference in the gelatin gel strength related
1.23 ± 0.09%. These valus met GMIA standard, to amino acid chain length, imino acid
ranging from 0.3 - 2.0% and the maximum value composition (prolin and hydroxyprolin), the
is 2.6% recommended by Muyonga et al. (2004) concentration of gelatin and the distribution of
The highest ash content found in the treatment at molecule weight (Arsenen and Gildberg, 2007;
the HCl concentration of 0.3 M, and then Karim and Bhat, 2009). Gelatin with long chains
decreasing at the HCl concentration of 0.6M had high gel strength values. The values have met
and 0.9M. Low ash content is indicative of the GMIA standard (2012), which were 50-300 g
good quality gelatin extraction process (Uriarte et bloom to be applicable in general industry.
al., 2011; Chandra and Shamasundar et al., 2015).
Degree of acidity (pH)
Gel Strength The HCl concentration significantly effected
Gel strength is a key parameter in the (P<0.05) on gelatin acidity (Table 1). The Increasing
determination of gelatin quality. The gel HCl concentration, degreased in pH of gelatin, at
strength indicates the capability of gelatin to 1.2 M and 1.5M of HCl concentration, the pH was
change from gel phase into sol phase and vice unaffected significantly. The higher HCl
versa (Kusumawati et al., 2008). The value of concentration caused a progressive increase in the
buffalo hide gelatin at the HCl concentration of number of hydrogen ions (Kusumawati et al.,
0.3 M to 1.5 M was in the range of 239.44 - 2008). The pH value at the HCl concentrations of
293.41 g bloom. 0.3 M to 1.5 M was in the range of 5.08 – 5.52. It
The results of the measurement of the gel still met industrial standard (GMIA, 2012), which
strength described that the HCl concentrations were 3.8 – 6.0.
had significant impact (P<0.05) on the gel
strength (Table 1). The HCl concentration of 1.2 Viscosity
M showed the highest gel strength value The viscosity of the buffalo hide gelatin
(293.41±0.73 g bloom), while the gel strength were in the range of 16.37 to 22.17 cp. The values
value at the HCl concentration of 1.5 M decreased have met GMIA quality standard (2012) for food
to the same value those found at the HCl industry, which were 1.5 – 7.5 cp. The HCl
concentrations of 0.9 and 0.6 M. The concentration gave significantly effect (P<0.05)
concentration of pretreatment material had impact on the viscosity. One of the influencing factors of
on the gelatin gel strength. High concentration of the gelatin viscosity is pH value (Kusumawati et
acid could cause a decrease in the gel strength al., 2008). Considering the pH values summarized
(Ockerman and Hansen, 2000). At the in Table 1, the pH of the buffalo hide gelatin was
concentration of 1.2 M, HCl brokedown amino in the range that still met industrial quality
acid polymer chain at the right limit. This standard and optimal pH for gelatin production.

18 J.Indonesian Trop.Anim.Agric. 42(1):14-22, March 2017

Jamilah and Harvinder (2002) suggested that the (more higher HCl concentration), this was still in
viscosity of the gelatin could be increased by the expected limit. There was an increase in the
extracting the gelatin at the pH of 3-10.5. In the red color intensity at 0.6 M of HCl concentration
range of the optimum pH for collagen hydrolysis as compared to 0.3 M, but this was not
(the breaking of the collagen chain took place in significantly different (P>0.05) with 0.9 M,
limit of polypeptide) the protein chains are still 1.2M, and 1.5M. The higher collagen hydrolysis
long and the gelatin viscosity was high (Said caused the increasing supply of free amino group.
et al., 2011; Pelu et al., 1998). The amino group reacts to the skin carbonyl
compound is non- enzymatic browning reaction
Color this cause the increase in the redness and
The color and the lightness of the gelatin yellowness of the gelatin (Nagarajan et al.,
present esthetic characteristics influencing the 2012; Sae-Law and Benjakul, 2015; Sae-Law et
acceptability and the applicability of the gelatin. al., 2016).
In general, the color of the gelatin did not have
any significant impact on its function (Table 1). Fourier Transform Infrared (FTIR)
However, more light colors are more liked Spectroscopy
because they provided more flexibility to be The FTIR is applied to observe the change in
mixed into certain food system (Rahman and the functional group and the secondary
Jamalulain, 2012; Shyni et al., 2014). The results structure of the gelatin (Muyonga et al., 2004;
of the measurement of the colors of the buffalo Kaewruang et al., 2013). Figure 1 shows the
hide gelatin were expressed in brightness values FTIR spectra of buffalo hide gelatin through the
(L*), redness intensity (a*) and yellowness pretreatment with HCl at the various
intensity (b*). The HCl concentration did not have concentrations from 0.3 to 1.5M. Muyonga et al.,
any significant impact (P>0.05) on the brightness (2004) reported that there were four identified
value or on the yellowness intensity, but it amide areas in the wave numbers of 3600 – 2300
had significant impact (P<0.05) on the intensity of per cm (amide A), 1656 – 1644 per cm (amide I),
the red color of the gelatin. The use of the HCl 1560-1335 per cm (amide II) and 1240 – 670 per
concentrations of 0.3 – 1.5 M was still in the safe cm (amide III).
limit for pretreatment process. This means that The amide A of the buffalo hide gelatin at
though the hydrolysis process proceeding higher the HCl concentrations of 0.3 M, 0.6 M, 0.9 M,
with the increase in the concentration of H+ 1.2 M and 1.5 M were respectively detected in the

Figure 1. FTIR Spectra of Buffalo Hide Gelatin by Pretreatment using HCl

Buffalo Hide Gelatin by HCl Pretreatment (S. Mulyani et al.) 19

wave numbers of 3402.43 per cm, 3425.58 helix transformation into coil random structure
per cm, 3425.58 per cm, 3448.72 per cm and may take place more frequently with the increase
3425.58 per cm. The peak of the amide A in the acid concentration of 0.9 M. The change
related to the stretch of the group N-H and was related to the loss of the triple helix structure as a
indicative of the existence of hydrogen bond. result of collagen denaturizing into gelatin
Vibration normally take place in the wave (Muyonga et al., 2004; Nagarajan et al., 2012).
numbers of 3400 – 3440 per cm. The lower wave
numbers indicated bigger molecular degradation CONCLUSION
of the gelatin. The degradation produces free
amino group that interracts with other reactive The difference in HCl concentrations had
groups (Kaewruang et al., 2013). The gelatin at impact on the yield, the characteristics and the
the HCl concentration of 1.2M had amide A peak secondary structure of the buffalo hide gelatin.
with the highest wave number (Figure 1). In other The HCl concentration of 0.9 M represented the
words, the molecular degradation was the optimum concentration to produce the highest
smallest with the longest molecular chain that the yield. In general, the characteristics of the
viscosity value and the strength of the gel were buffalo hide gelatin with the pretreatment process
the highest. using HCL had the characteristics that met the
The amide I presents the stretch C=0 or GMIA standard.
hydrogen bond in a pair with COO. The absorbing
area of the amide I is often used to analyze REFERENCES
protein secondary structure (Nagarajan et. al.,
2012). The amide I of the buffalo hide gelatin at Lee, J.W., S.B. Choi, Y.H. Jung, J.F. Keown and
the HCl concentrations of 0.3 M, 0.6 M, 0.9 M, L.D. Van Vleck. 2000. Parameter estimates
1.2 M and 1.5 M, respectively were detected in for direct and maternal genetic effects on
the same wave number of 1635.64 per cm. yearling, eighteen-month, and slaughter
Yakimates et al. (2005), Nagarajan et al. (2012) weights of Korean native cattle. J. Anim.
stated may represent the peak absorption took Sci. 78:1414-1421.
place in the wave number of 1633 per cm Ahmad, M and S. Benjakul. 2011. Characteristics
representing the characteristic of the coil structure of gelatin from the skin of unicorn
of the gelatin. Concerning with transmission leatherjacket (Aluterus monoceros) as
intensity, the gelatin at the highest HCl influenced by acid pretreatment and
concentration of 0.9 M experienced triple helix extraction time. Food Hydrocoll. 25:281-288
dispersal into coil structure whose quantity was Alfaro, A.T., F. C. Biluca, C. Marquetti, I.B.
the highest. Therefore, the pretreatment with the Tonial and N.E.de Souza. 2014. African
HCl at the concentration of 0.9 M produced the catfish (Clarias gariepinus) skin gelatin:
highest yield (Table 1). Extraction optimization and physical-
The amide II of the buffalo hide gelatin at chemical properties. Food Res. Int. 65:416-
the HCl concentrations of 0.3 M, 0.6 M, 0.9 M, 422
1.2 M, and 1.5 M was detected in the range of the AOAC. 2005. Official Methods of Analyses
wave numbers of 1527.62 – 1342.46 per cm Association (18th Ed.) Association of
(Figure 1). The absorption of the amida II takes Analytical Chemist, Washington, D.C
place because of the vibration of the N-H group Arsenen, J.A. and A.Gildberg. 2007. Extraction
and the stretch of the group C-N (Nagarajan et al., and characterization of gelatin from Atlantic
2012). The amide III was detected in the wave salmon (Salmo salar) skin. Bioresour.
numbers of 1242.16 – 871.82 per cm. This Technol. 98 (1):53-57
showed the combination of the stretch peak of Benjakul, S., K. Oungbho, W. Visessanguan, Y.
C-N and the deformation of N-H in the amide Thiansilakul, and S. Roytrakul. 2009.
was observed as the absorbance of the group Characteristic of gelatin from the skins of
CH2 of the glycine backbone chain and the bigeye snapper, Priacanthus tayenus and
proline side chain. The gelatin at the HCl Priachantus macracanthus. Food Chem.
concentration of 0.9 M was indicative of the 116:445-451
highest transmittance intensity for both amide II Binsi, P.K., B.A. Shamasundar, A.O. Dileep, F.
and III. T h i s indicated that the bigger random Badii and N.K. Howell. 2009. Rheological
molecular structure may be exist because the a- and functional properties of gelatin from the

20 J.Indonesian Trop.Anim.Agric. 42(1):14-22, March 2017

 skin of Bigeye snapper (Priacanthus 63-68
hamrur). Food Hydrocoll. 23: 132-145. Muyonga, J.H., G.G.B. Cole and K.G. Duodo.
Chandra, M.V. and B.A. Shamasundar 2015. 2004. Extracting and physicochemical
Rheological properties of gelatin from swim characterization of Nile perch (Latec
bladders of freshwater fish Catla catla. Food niloticus) skin and bone gelatin. Food
Hydrocoll. 48: 47-54 Hydrocoll.18:581-592
Eastoe, J.E. and AA. Leach 1977. Chemical Nagarajan, M., S. Benjakul, T.Prodptan, P.
Constitution of Gelatin. In: Ward AG, Courts Songtipya and H. Kishimura. 2012.
A. editors. The Science and Technology of Characteristic and functional properties of
Gelatin. Academic Press, New York. gelatin from splendid squid (Loliga
GMIA. 2012. Gelatin Handbook. Gelatin formasana) skin as affected by extraction
Manufacturers Institute of Amerika temperature. Food Hydrocoll. 20: 389-397
Haug, I.J. and K.I. Draget. 2009. Gelatin in Nikoo, M., S. Benjakul, M. Bashari, M.
Handbook of Hydrocolloids. Norwegian Alekhorshied, A.I. Cissouma, N. Yang and
University of Science and Technology X. Xu. 2014. Physicochemical properties of
(NTNU), Norway skin gelatin from farmed amur sturgeon
Huda, N., A.A. Putra and R. Ahmad. 2011. (Acipenser schrenckii) as influenced by acid
Physicochemical and nutritional pretreatment. Food Biosci.5:19-26
characteristics of Indonesian buffalo skin Niu, L., X. Zhiou., C. Yuan., Y. Bai., K., Lai.,
crackers. Int. J. Meat Sci. 1 (1): 36-51 F.Yang and Y. Huang. 2013. Characterization
Jamilah, B and K.G. Harvinder. 2002. Properties of tilapia (Oreochromis niloticus) skin
of gelatins from skins of fish black tilapia gelatin extracted with alkaline and different
(Oreochromis mossambicus) and red tilapia acid pretreatments. Food Hydrocoll. 3:336-
(Oreochromis nilotica). Food Chem. 77:81- 341
84 Ockerman, H.W and G. L. Hansen. 2000. Animal
Johnston-Bank, F.A. 1990. From Tannery to By-Product Processing and Utilization.
Table: An account of gelatin production. J. CRC-Press
of the Soc. of Leather Technol. and Chem. Pelu, H., S. Harwanti and E. Chasanah. 1998.
68:141-145. Ekstraksi gelatin dari kulit ikan tuna melalui
Kaewruang, P., S, Benjakul, T. Prodpran, and S. proses asam. J. Penelitian Perikananan
Nalinanon. 2013. Physicochemical and Indonesia. 4(2):66-74.
functional properties of gelatin from the skin Pranoto, Y., M.L. Chong and H.J. Park. 2006.
of unicorn leatherjacket (Aluterus Characterization of fish gelatin film added
monoceros) as affected by extraction with gellan and κ-Carragenan. LWT- Food
condition. Food Bio Sci. 2: 1-9. Sci. and Technol. 40:766-774.
Karim, A.A. and R. Bath. 2009. Fish Gelatin; Rahman, M.N.A and S.A.S.K.A., Jamalulail.
Properties, Chalenges and Prospect as an 2012. Extractions, physicochemical
alternative to mammalian gelatin. Food characterizations and sensory quality of
Hydrocoll. 23: 563-576. chicken feet gelatin. Borneo Sci. 30:1-13.
Kolodziesjka; E. Skierka., M.Sadowska., W. Said, M.I., J.C. Likadja and M. Hatta. 2011.
Kolodziesjka and C. Nieckowska. 2007. Pengaruh waktu dan konsentrasi bahan
Effect of extracting time and temperature on curing terhadap kuantitas dan kualitas
yield of gelatin from different fish offal. gelatin kulit kambing yang diproduksi
Food Chem. 107:700-706. melalui proses asam. Jurnal Ilmu dan
Ktari. N., I. Bkhairia, M, Jridi, I. Hamza, B.S. Teknologi Peternakan. 1:119-128
Riadh and M. Nasri. 2014. Digestive acid Sae-Leaw, T and S. Benjakul. 2015.
protease from Zebra blenny (Salaria Physicochemical properties and fishy odour
basilisca): Characteristics and application in of gelatin from seabass (Lates calcarifer)
gelatin extraction. Food Res. Int. 57:218-224 skin stored in ice. Food Bio Sci. 10: 59-68
Kusumawati, R., Tazir and A. Wawasto. 2008. Sae-Leaw, T., S.Benjakul, and N.M. O’Brien.
Pengaruh Perendaman dalam Asam Klorida 2016. Effect of defatting and tannic acid
terhadap kualitas Gelatin Tulang Kakap incorporation during extraction on properties
Merah (Lutjanus sp). J. Pasca Panen dan and fishy odour of gelatin from seabass skin.
Bioteknologi Kelautan dan Perikanan. 3(1): LWT-Food Sci. and Technol. 65:661-667.

Buffalo Hide Gelatin by HCl Pretreatment (S. Mulyani et al.) 21

 See, S. F., P.K. Hong, K.L. Ng, W.M. Wan Aida Spanghero, M., L.Gracco, R. Valusso and
and A.S. Babji. 2010. Physicochemical Piasentier. 2004. In vivo performances,
properties of gelatin extracted from skins of slaughtering traits and meat quality of
different freshwater fish species. Int. Food bovine (Italia simmental) and buffalo (Italian
Res. J. 17:809-816. Mediterranean) bulls. Livest. Prod. Sci.
Shyni, K., G.S. Herma, G. Ninan, S. Mathew, S., 91:120-141.
Joshy and P.T. Lakshmanan. 2014. Isolation Uriarte-Montoya, M.H., H. Santacruz-Ortega,
and characterization of gelatin from skins of F.J.Cinco-Moroyoqui, O. Rouzaud-Sandez,
Skipjack tuna (Katsuwonus pelamis), dog M.Plascencia-Jatomea and J.M.Ezquerra-
shark (Scoliodon sorrakowah), and rohu Brauer. 2011. Gian squid skin gelatin:
(Labela rohita). Food Hydrocoll.39: 68-76 Chemical composition and biophysical
Sithusamran, S., S. Benjakul and H. Kishimura. characterization. Food Res. Int. 44:3243-
2014. Characteristics and gel properties of 3249
gelatin from skin of Seabass (Lates Wang, L., A.M. Rau, J.F. Kerry, and J.P. Kerry.
calcarifer) as influenced by extraction 2008. Effect of pH and addition of corn oil
condition. Food Chem. 152:276-284. on the properties of gelatin based
Sompie, M., S.E. Surtijono, J.H.W. Pontoh and biopolymer film. J. Food Eng. 90:11-19.
N.N. Lontaan. 2015. The Effects of acetic Yakimates, I., N.Wellner, A.C. Smith,
acid concentration and extraction R.H.Wilson, I.Farhat and J. Mitchell. 2005.
temperature on physical and chemical Mechanical properties with respect to water
properties of pigskin gelatin. Proced. Food content of gelatin films in glassy state.
Sci. 3: 383-388. Polym. 46:12577-12585.

22 J.Indonesian Trop.Anim.Agric. 42(1):14-22, March 2017

View publication stats