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Extraction and characteristic of Dioscorea alata mucilage

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AESAP 2019 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 542 (2020) 012016 doi:10.1088/1755-1315/542/1/012016

Extraction and characteristic of Dioscorea alata mucilage

D Fortuna1, S S Mardjan2, T C Sunarti3, E Darmawati2, S M Widayati4, and N

Purwanti2,5
1
Doctoral Program in Agricultural Engineering Science, Department of Mechanical and
Biosystem Engineering, Bogor Agricultural University, Bogor Indonesia
2
Department of Mechanical and Biosystem Engineering, Bogor Agricultural University,
Bogor Indonesia
3
Department of Agricultural Industry Technology, Bogor Agricultural University, Bogor
Indonesia
4
Indonesia Center for Agricultural Postharvest Research and Development (ICAPRD), Bogor,
Indonesia
5
Corresponding author, E-mail: nanik_purwanti@apps.ipb.ac.id

Abstract. Dioscorea alata (DA) mucilage is classified as water-soluble polysaccharides,

which has potential as a source of hydrocolloid. This research was conducted to investigate
effects of salt types on water to tuber ratio to produce the most optimal mucilage yield. This
research was conducted using Completely Randomized Factorial Design 3 x 5. The DA
mucilage was extracted using two factors, i.e., types of salt (without salt, sodium chloride, and
calcium chloride) and water to tuber ratios (2:1, 4:1, 6:1, 8:1, and 10:1). The results showed
that the types of salt significantly affect (p<0.05) on starch content, but did not have a
significant effect on mucilage yield and water content of the mucilage. The water to tuber
ratios significantly affected (p<0.05) the mucilage yield and starch content of the mucilage,
however it did not affect the water content. Addition of calcium chloride when water to tuber
ratio was 4:1 was recommended to produce the optimal mucilage yield (1.58%) with relatively
low starch content (7.86%).

1. Introduction

Hydrocolloids are hydrophilic polymers derived from various sources such as plants, animals,
microorganisms or chemical modification, that have multi functionalities such as thickener, gelling
agent, stabilizer, etc. The world market of hydrocolloids was estimated to reach USD 8.77 billion
(around IDR 119.9 trillion) in 2018, but price instability and shortage of raw materials were
considered to be obstacles in hydrocolloids market [1].
Dioscorea sp. is one of the tubers belongs to Dioscoreaceae family that has a potential as
hydrocolloids source because of its mucilage content. The viscous mucilage mainly contains protein
and polysaccharides in the form of glycoprotein, a mannan that is firmly associated with protein [2].
Dioscorea mucilage is water soluble carbohydrates-protein complexes with a ratio between soluble
carbohydrates and protein of 1:8.1. Monosaccharide composition in mucilage of Dioscorea alata
(DA) mainly composed of mannose (95.4%). Other units like glucose, galactose, arabinose, xylose,
and rhamnose also present at low quantities, i.e., below 5% [3]. A previous research showed that
mucilage yield extracted from DA is higher than that of taro and sweet potato. On dry basis, DA
produced 6.51 g of mucilage (/100 g material) compared to 3.23 g per 100 g taro and 5.12 g per 100 g
sweet potato [4]. Rehydrated mucilage of DA can be used to produce oil-in-water or air-in-water
emulsion (foam) even at concentration of 0.1% with good stability. Emulsions with 0.5% DA
mucilage was reported to have soft creamy and thixotropic semisolid texture [5]. In addition, DA

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IOP Conf. Series: Earth and Environmental Science 542 (2020) 012016 doi:10.1088/1755-1315/542/1/012016

mucilage has antioxidant, antiradical, and immunostimulatory activity. DA mucilage var. purpurea
(Roxb.) Ming-Jen has an inhibition concentration (IC50) of 0.547 mg/ml [6,7].
Several studies on mucilage extraction from Dioscorea sp. have been done using water as the
solvent. Ratio of water to solid material is one of the main factors during extraction [6]. Some studies
used water to solid ratios that vary from 0: 1 to 10: 1 [9,10]. The mucilage yield increased with
increasing water to solid ratio before it turned to be constant and then, decreased at a certain point.
The ratio of water to solid plays an important role in reducing resistance to mass transfer during
extraction. Enzymes can be activated to facilitate separation of protein and starch from the mucilage
by increasing water to solid ratio. However, the optimal ratio must be defined to minimize extraction
cost [10].
One of the challenges in extracting mucilage from DA is high viscosity and high water-binding
capacity of glycoprotein that inhibit separation of mucilage from starch [3]. Salts have been widely
used to release mucilage during starch extraction from various plants [10]. Sodium chloride (NaCl) at
a concentration of 1% was used to remove mucilage during starch extraction of Dioscorea tuber [11].
Addition of 0.3 M NaCl was reported to be effective in removing mucilage from taro tuber [12]. The
addition of NaCl with concentrations of more than 0.5% reduces water holding capacity; therefore,
mucilage would be easier extracted from the tubers [13].
This research was performed to investigate effects of salt types on water to tuber ratio during
mucilage extraction from Dioscorea alata tuber to produce the most optimum mucilage yield. The
experiments were carried out using two salts commonly used in food industry, i.e., NaCl (monovalent
salt), and calcium chloride (CaCl2, divalent salt).

2. Materials and methods

2.1 Materials and chemicals

Dioscorea alata (yellow yam) was harvested from one of the farming complexes within Bogor
Agricultural University. The tubers were harvested after their stems withered, which approximately
12 months after planting.
Analytical grade reagents used in this study were absolute ethanol (Cat. No. 1009835000, Merck,
Germany), NaCl (Cat. No. 1064041000, Merck, Germany), CaCl2 (Cat. No. 1023780500, Merck,
Germany) and distilled water.

2.2 Extraction of mucilage from Dioscorea alata tuber.

Fresh tubers were washed, peeled, and sliced around 1 cm thickness. Sliced tubers (250 g) were steam
blanched for 10 min and further blended for 10 min in 1% w/w of salt solutions, with different water
to tuber ratios. The slurry was stored at 10C for 24 hours and then, filtered through a 105m sieve.
Further, the filtrate was centrifuged (SorvallTM LegendTM XTR centrifuge, Fisher Scientific Ltd, UK)
at 12,000g for 20 min at 4C from which the supernatant was collected. The supernatant was mixed
with ethanol with a ratio of 1:1 (v/v) and kept at 4C for 24 hours and then, centrifuged at 12,000g for
20 min at 4C to precipitate the mucilage. The precipitate was freeze-dried and then, ground to
obtained mucilage powder. The powder was stored at room temperature for further analysis.
The experiments in this research was a 3 × 5 factorial in Completely Randomized Design. The
independent variables were three types of salt (control/water, NaCl, and CaCl2) and five of water to
tuber ratios (2:1, 4:1, 6:1, 8:1, and 10:1). The experiments were carried out in duplicates. The data
were evaluated for their significant differences by Duncan’s Multiple Range Test (DMRT) (p = 0.05).

2.3 Chemical composition of fresh Dioscorea alata tubers

The moisture and crude ash were determined using gravimetric method, crude fat content was
analysed using soxhlet extraction method, crude protein content was measured using Kjeldahl
method, and carbohydrate content was calculated by difference method [3,8]. Starch content was
determined using Luff Schoorl method. Each method of analysis was performed in triplicate.

2.4 The mucilage yield

The mucilage yield was expressed as ratio (% w/w, YM) between the weight of extracted mucilage (A)
and the initial weight of fresh tuber (B) [14], as shown in equation (1).
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AESAP 2019 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 542 (2020) 012016 doi:10.1088/1755-1315/542/1/012016

(1)

2.5 Starch content of the mucilage

The starch content of DA mucilage was determined using Megazyme’s Total Starch (-
amylase/Amyloglucosidase, AA/AMG) Assay kit (K-TSTA) [8]. This analysis was performed to
check purity of the mucilage.

2.6 Mucilage morphology

The morphology of dried mucilage was observed using an environmental scanning electron
microscope (SEM) (FEI Quanta 650, FEI Company, Hillsboro, Oregon, USA). The samples were
placed on carbon conductive tape without coating. Then, the scanning was operated at low vacuum
mode using a Large Field Detector (LFD) at 500× magnification, 10 mm of working distance, with
pressure, voltage and temperature of 60 Pa, 10 kV, and 20C respectively.

3. Results and discussion

3.1 Chemical composition of fresh Dioscorea alata tubers

Chemical composition of the fresh tubers (without peel) is stated in table 1. On a wet basis, moisture
was the main content of fresh tubers. On a dry basis, the highest content was carbohydrate, followed
by starch and then, protein. The values presented in table 1 are in accordance with previous studies
that reported varying chemical contents of tubers, i.e., 15.24 – 70.88% of carbohydrate, 14.25 – 77.40
% of starch, and 2.86 – 13.20% of protein [9,10,15,16]. These results confirm previous researches that
DA mucilage mainly contains carbohydrates and protein. This composition might lead to high viscous
slurry that makes separation process becomes challenging.
Table 1. Chemical composition of fresh Dioscorea alata tubers

Parameter % dry basis % wet basis

Moisture - 61.65
Crude Ash 2.76 1.06
Crude Fat 0.31 0.12
Protein 32.41 12.43
Carbohydrates 64.51 24.74
Starch 44.82 17.19

3.2 Mucilage yield

This research indicated that types of salt did not significantly affect the mucilage yields (p>0.05), as
shown in table 2. On the other hand, water to tuber ratios significantly affected the yields (p>0.05).
There were no interaction effects between types of salt and water to tuber ratios. The result shows that
the mucilage yield increased with increasing water to tuber ratios, and the highest yield was obtained
when water to tuber ratio was 6:1. Increasing the ratio beyond this resulted in decreasing mucilage yield.
Nevertheless, this research resulted in 10× higher mucilage yield than a previous study [9] at the highest
water to tuber ratio (10:1). This might be attributed to centrifugational process of filtrate during
separation.
The mucilage yield increased with increasing ratio of water to tuber. At high water to tuber ratio,
more volume of water is available to penetrate and diffuse through the tuber matrices. More water
presents would bind more water-soluble compounds in the tuber cells, including the mucilage, and
they are easier to be extracted from the cells. A higher volume of water presents during extraction
would also make less viscous slurry, therefore, separation of water-soluble compounds from the
tubers is more efficient. This particular results are in line with previous studies that reported a higher
yield of mucilage extracted from various Dioscorea sp. [17,18,19].
Table 2. The mucilage yield (%) affected by types of salt and
water to tuber ratios
The type of salts Mucilage yield (%)

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AESAP 2019 IOP Publishing
IOP Conf. Series: Earth and Environmental Science 542 (2020) 012016 doi:10.1088/1755-1315/542/1/012016
Water 1.47
NaCl 1.46
CaCl2 1.31
The water to tuber ratio Mucilage yield (%)
2:1 0.58 a
4:1 1.58 bc
6:1 2.06 c
8:1 1.46 b
10:1 1.40 b
Means with the same superscript in the same column are not
significantly different (p>0.05)
Salt addition was expected to have positive effects in the mucilage extraction from the tubers,
which is not significantly observed in table 2. The addition of salt causes osmotic pressure, which
tends to push water together with all water-soluble compounds out of the cells. The addition of NaCl
also causes an increase in ionic strength, affects hydrogen bonds and ionic bonds, changes the higher-
order structure of mucilage; therefore, the mucilage viscosity decreases due to increased surface
hydrophobicity [19]. Another author described that the apparent viscosity of Mesona blumes gum is
lower with addition of CaCl2 than NaCl [20]. This was also inline with another study on viscosity of
Dicerocaryum zanguebarium mucilage that decreased steadily when CaCl2 or NaCl was added [20].
Nevertheless, salts can be attributed to the loss of the hydrophilic–hydrophobic balance of the
polymer networks [21], which might affect water holding capacity of polysaccharides [22]. This
might be the reason for insignificant effects of salt addition to the mucilage yields as shown in table 2.

3.3 Starch content of mucilage

Table 3 shows that water to tuber ratios, types of salt, and their interactions had significant effects
(p>0.05) on starch content in the mucilage. Different water to tuber ratios did not significantly affect
the starch content in the mucilage when water did not contain any salts. When either NaCl or CaCl2
was added, the results can be devided into two groups of starch content. The group with low starch
content was attributed to water to tuber ratio of 2:1 and 4:1; meanwhile, the group with high starch
content was attributed to water to tuber ratios higher than 4:1. This research aimed for as low starch
content in the mucilage as possible, although the presence of starch in the mucilage was inevitable.
Overall, starch contents in the mucilage obtained in this research were much lower that the value
reported previously by Alves et al. [9] which was about 43%.
Table 3. Starch content (%) of mucilage extracted with various water to tuber
ratios and different salts.
Types of salt
Water to tuber ratio
Water NaCl CaCl2
2:1 16.99 a A 14.04 a A 4.42 b A
4:1 17.40 a A 17.82 a AB 7.86 b A
6:1 19.64 a A 18.78 a B 16.62 a B
8:1 19.79 a A 18.18 a B 15.44 a B
10:1 18.77 a A 18.68 a B 14.54 a B
Means followed by the same letter do not significantly different by Duncan’s test (p>0.05)
A–B
Values with different superscripts within the same column significantly different
(p<0.05).
a–b
Values with different superscripts within the same row significantly different (p<0.05).

The mucilage with the lowest starch content was produced using water to tuber ratio lower than 4:1
added with CaCl2 (table 3). It is still not clear from the results how salts affected starch content in the
mucilage, especially with low ratio of water to tuber. However, it has been known that salts encourage
breakdown of starch, i.e., depolymerization of glucose chains into smaller molecules [23] and affect
starch gelatinization. When starch is gelatinized, other molecules such as proteins are easier to be
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IOP Conf. Series: Earth and Environmental Science 542 (2020) 012016 doi:10.1088/1755-1315/542/1/012016

extracted. The gelatinization inhibition effect of Ca2+ is lower than that of Na+ with concentrations of
less than 1 M. Protein solubility can also increase when Ca2+ presents rather than Na+ [24]. These
might lead more effective separation process, therefore, higher purity of mucilage.

3.4 Mucilage morphology

figure 1 and figure 2 show morphologies of DA mucilage powder captured using SEM at 500× and
5000× magnification, respectively. SEM was used to obtain three-dimensional information on the
effects of the types of salt on surface morphology of DA mucilage particles. The surface morphology
and structure of mucilage particles are affected by extraction, purification, and preparation methods
[30]. This research shows that types of salt affected porosity of DA mucilage particles. Without salt,
particles of DA mucilage were fragmented with slightly porous surface and irregular shape (figure 1a
and figure 2a). Particles of DA mucilage with NaCl salt were dense with smooth surface (figure 1b
and figure 2b). Last, addition of CaCl2 during mucilage extraction produced mucilage particles with
porous structure (figure 1c and figure 2c). A research indicated that high concentration of salts affects
starch gelatinization and surface morphology of the starch from various tubers [31]. However, further
studies are required to explain salt effects on morphology of dried mucilage.

(a) (b) (c)

Figure 1. Scanning electron microscope image of Discorea alata mucilage (The water to tuber ratio
of 6:1) at magnifications of 500x (a) without salt, (b) NaCl, (c) CaCl2

(a) (b) (c)

Figure 2. Scanning electron microscope image of Discorea alata mucilage (The water to tuber ratio
of 6:1) at magnifications of 5000x (a) without salt, (b) NaCl, and (c) CaCl2

4. Conclusions

This research shows that water to tuber ratios used during extraction of DA mucilage affected the
yield. The ratio together with addition of salt affected starch content as well as morphology of
mucilage particles. Water to tuber ratio of 4:1 with addition of CaCl2 salt resulted in mucilage with
the optimal yield (1.58%) and high purity (low starch content). When NaCl was added, water to tuber
ratio of 6:1 must be applied to get the highest yield of mucilage (2.06%).

Acknowledgments
The research was financially supported by Beasiswa Unggulan Dosen Indonesia Dalam Negeri
(BUDI-DN), The Indonesia Endowment Fund for Education (LPDP), Ministry of Finance Republic of
Indonesia, and Ministry of Research, Technology and Higher Education of the Republic of Indonesia.

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IOP Conf. Series: Earth and Environmental Science 542 (2020) 012016 doi:10.1088/1755-1315/542/1/012016

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