Journal of King Saud University – Engineering Sciences (2014) 26, 3–9

King Saud University

Journal of King Saud University – Engineering Sciences

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ORIGINAL ARTICLE

Effect of process conditions on the gel viscosity

and gel strength of semi-refined carrageenan (SRC)
produced from seaweed (Kappaphycus alvarezii)
Awang Bono *, S.M. Anisuzzaman 1, Ong Wan Ding 2

Chemical Engineering Programme, School of Engineering and Information Technology (SEIT), Universiti Malaysia Sabah
(UMS), 88400 Kota Kinabalu, Sabah, Malaysia

Received 7 February 2012; accepted 20 June 2012

Available online 28 June 2012

KEYWORDS Abstract Kappaphycus alvarezii or commonly known Euchema cottonii is a good source of kappa-
Euchema cottonii; carrageenan and can be found cultivated in the coastal areas of Malaysia, Philippines and Indone-
Rhodophyceae; sia. Carrageenans have many applications and are utilized in human food and pet-food industry.
Alkali treatment; Carrageenans are also utilized in non-food industry such as pharmaceuticals, cosmetics, printing
Cooking time; and textile formulations. Currently, the Southeast Asian region is producing semi refined carra-
Cooking temperature geenan (SRC). There are various works in producing SRC; however, there are limited efforts to
develop the optimization of cooking process parameters. Hence, the present study features on
the cooking process (alkaline treatment) where the parameters (concentration of potassium hydrox-
ide solution, cooking time and cooking temperature) and the ranges are identified experimentally.
The effects of these parameters on carrageenan quality such as gel viscosity and gel strength were
studied. The optimization of cooking process parameters and the experimental design was con-
ducted based on the Central Composite Design (CCD) of Response Surface Methodology
(RSM). The experimental result showed that gel viscosity increases with the decrease of cooking
time, cooking temperature and potassium hydroxide (KOH) concentration (% w/w). In contrast,
gel strength increases as cooking time, cooking temperature and KOH concentration (% w/w)
increases. From the optimization, the best conditions for alkaline treatment found were cooking

* Corresponding author. Tel.: +60 88 320000x3027, +60 16

4150102; fax: +60 88 320348.
E-mail addresses: bono@ums.edu.my (A. Bono), anis_zaman@
ums.edu.my, anisbg2001@yahoo.com (S.M. Anisuzzaman),
ding2487@gmail.com (O.W. Ding).
1
Tel.: +60 88 320000x3222, +60 16 8237107; fax: +60 88 320348.
2
Tel.: +60 12 8872787; fax: +60 88 320348.
Peer review under responsibility of King Saud University.

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1018-3639 ª 2012 Production and hosting by Elsevier B.V. on behalf of King Saud University.
http://dx.doi.org/10.1016/j.jksues.2012.06.001
4 A. Bono et al.
temperature 80 C, cooking time 30 min and KOH concentration 10 (% w/w) which are similar to
current practice in industry.
ª 2012 Production and hosting by Elsevier B.V. on behalf of King Saud University.
1. Introduction
Carrageenan is a generic name for polysaccharides, extracted
from different species of red seaweeds (Rhodophyceae); for in-
stance, Gigartina, Chondruscrispus, Euchema and Hypnea
(Campo et al., 2009). Chemically, carrageenans are a family
of hydrophilic linear sulfated galactans found exclusively in
the cell walls of red marine algae (Rhodophyceae). The diverse
family of polysaccharides however shares a common galactan
backbone of alternating 3-linked-b-D-galactopyranose (G-
units) and 4-linked-a-D-galactopyranose (D-units). The
important families of these carrageenans are widely used in
industry. They include k-carrageenan (kappa, G4S-DA) and
i-carrageenan (iota, G4S-DA2S) which have the ability to form
thick solution or gels and k-carrageenan (G2S-D2S6S) which
have thickening abilities (Jouanneu et al., 2010).
Malaysia, Indonesia and Philippines particularly have suit-
able sheltered areas that are favorable for cultivation. k-Carra-
geenan is obtained through the extraction of tropical red
seaweed Kappaphycus alvarezii, better known in trade as
Euchema cottonii (Duduku et al., 2008; Campo et al., 2009).
K. alvarezii is the most important source of k-carrageenan in
the world and highly demanded for its cell wall polysaccharide
(Muñoz et al., 2004).
For over the past four decades, K. alvarezii has been shown
as an industrially important carrageenophyte (Bindu and
Levine, 2010). The Southeast Asian countries have produced
96.5% of the total production of k-carrageenan extracted from
red seaweed; of which 55% is contributed by Philippines, fol-
lowed by Indonesia (38%) and Malaysia (2.5%).
Carrageenan is inarguably one of the most useful resources.
Carrageenans are widely utilized in food industry because of
their physical and functional properties, such as thickening,
gelling and stabilizing abilities (Bono et al., 2011). Referring
to Arifin et al. (2003), carrageenans are used as stabilizer,
emulsifier, thickener, filler and gelling agent for dairy-based
products; for instance, cheese, ice-creams, yogurt, butter, sau-
sages, paste thickener and others. Carrageenan also helps to
improve the quality of meat products. The largest application
for semi-refined carrageenan is the usage in producing pet food
at about 5500 tonnes annually (McHugh, 2003).
In non-food products, carrageenans are used in pharmaceu-
tical, cosmetics, printing and textile formulation industries
(Campo et al., 2009). The physical properties of carrageenan
have been investigated widely where various works had been
reported on methods used to extract carrageenan (Gunning
et al., 1998; Hayashi et al., 2010; Hoffmann et al., 1995; Hung
et al., 2008; Muñoz et al., 2004; Montolalu et al., 2008; Ride-
out et al., 1998; Uy et al., 2005). However, there are limited ef-
forts to develop the optimization of process parameters in
alkaline treatment. Hence, the present study emphasizes on
the production of SRC by alkaline treatment where the process
parameters such as cooking temperature, cooking time and the
KOH concentration will be investigated and optimized using
Response Surface Methodology. The rheological properties
of k-carrageenan (K. alvarezii species); gel strength and viscos-
ity are studied to determine the best condition for the alkaline
treatment.
2. Methodology
Basically, the laboratory work was divided into three parts; the
first part of the experiment was preparation of SRC with
cooking and drying process. Second part was characterization
of SRC followed by optimization of the process parameters in
alkaline treatment using Response Surface Methodology
(RSM).
2.1. Preparation of SRC from K. alvarezii
2.1.1. Preparation of raw seaweed
Raw seaweed was prepared by sun drying the fresh seaweed
collected from farm. Sun dried seaweed was washed thor-
oughly with distilled water for two reasons; to remove contam-
inants and to standardize the moisture content of the seaweed.
Wet seaweed was then dried in oven at 60 C for 15–16 h to re-
move the excess moisture.
2.1.2. Cooking of seaweed
The dried seaweed was cooked in a batch cooker in potassium
hydroxide (KOH) solution. The cooking temperature was con-
trolled to be constant, whereas the KOH concentration and
cooking time were fixed to the desired value. The experimental
runs were conducted according to the experimental design
which will be explained in Section 2.3.
The cooked seaweed was then cooled down to room tem-
perature and washed with distilled water to remove the excess
KOH. These processes using seaweed was classified as SRC.
2.1.3. Drying of cooked seaweed
After washing, the wet SRC was then dried in oven at 60 C
for 24 h. The dried sample was then chopped and grinded be-
fore it was kept in an air tight container to prevent the effect of
moisture from the surroundings. The samples were kept in
container for the characterization of carrageenan analysis.
2.2. Characterization of carrageenan
2.2.1. Gel viscosity measurement
The viscosity measurement was carried out according to Muñoz
et al. (2004) work. SRC powder (1.5 g) was dissolved in 100 ml
hot distilled water (1.5% solution) at 90 C and dispersed with
agitation using a magnetic stirrer for 20 min. The sample was
allowed to stabilize in a water bath at 80–90 C for 15 min to
eliminate bubbles that form as a result of the heating process
at temperature close to boiling temperature.
Viscosity was determined with a Brookfield DV-E Low Cost
Digital Viscometer using spindle No. 5 (19 mm in diameter,
approximately 65 mm in length) and capable of rotating at
Effect of process conditions on the gel viscosity 5

cooking temperature (60–85 C), cooking time (30–180 min)

Table 1 Range of process parameters.
and KOH concentration (5–12% w/w). Thus the experiment
Factors Lower limit Upper limit was designed to use the range of process parameters as shown
Cooking temperature (C) 60 80 in Table 1.
Cooking time (min) 30 90 The experimental runs and the conditions were conducted
KOH concentration (% w/w) 6 10 as suggested by Design-Expert Software version 7 with Cen-
tral Composite Design (CCD) of RSM (Bono et al., 2008;
Zhang et al., 2007).
30 rpm. After six complete revolutions of the viscometer, viscom- The optimum conditions of cooking process parameters
eter readings were taken. Viscosity was expressed as centipoises were established using the procedure included in Design-Ex-
(cP). The maximum viscosity using spindle No. 5 at 30 rpm pert Software. The input to the procedure was the range of
was 1250 cP. If the viscosity is very high and exceeds the maxi- process parameters as used in the experimental runs, whereas
mum reading, the rotational speed is adjusted to a lower value the target responses were chosen as the maximized condition.
(<30 rpm) but the spindle number is still maintained. According
to Sen and Erboz (2010), each run needed eight replicates. 3. Results and discussion

2.2.2. Gel strength measurement
SRC solution was prepared by dissolving 1.5 g of SRC powder
to 100 ml of distilled water with continuous magnetic stirring
at 90 C for 20–30 min. The sample was allowed to stabilize
in a water bath at 80–90 C for 15 min to eliminate bubbles
that form. The viscous solution was poured into three 50 ml
beakers (each of height 22 ml) moulded and left to cool for
20–30 min before sealing them. Samples were stored for 24 h
and maintained at ambient temperature (28 C) prior to anal-
ysis (Thrimawithana et al., 2010).
TA-XT plus texture analyzer (Stable Micro Systems,
England) was used to evaluate the compressibility of
carrageenan produced. A Load Cell of 5 kg was used where
the maximum force is ±5 kg. A fixture and an analytical probe
(P 0.5/R = 12.7 mm diameter) were compressed into the sam-
ple to a depth of 15 mm at a cross-head speed of 2 mm s1.
Each run was replicated three times.
2.3. Experimental design and optimization
Based on previous research works (Higuera et al., 2007)
showed, ranges of alkali treatment process parameters were
Based on the experimental design suggested by Design-Ex-
pert Software version 7, 20 runs of experiments were con-
ducted. The results are presented in Table 2.
The ANOVA analysis depicted that the experimental re-
sults can be fitted into a model, as shown in the following
Eqs. (1) and (2) in terms of actual factors like cooking temper-
ature (A), cooking time (B) and KOH concentration (C).
Gel Viscosity ¼ 2426:10  47:62A þ 59:89B  336:48C
 0:46AB þ 8:16AC  3:91BC ð1Þ
Gel Strength ¼ 179:31  2:42A  1:83B  29:57C
þ 0:02AB þ 0:49AC þ 0:04BC ð2Þ
3.1. Influence of process parameters on quality of SRC
3.1.1. Effect of temperature, time and KOH concentration upon
gel viscosity of SRC
Fig. 1 shows the properties of gel viscosity produced with var-
ious cooking times and cooking temperatures. It could be seen
that the gel viscosity clearly decreased with the cooking time,

Table 2 Experimental results.

Run Factor 1A: Cooking Factor 2B: Cooking Factor 3C: Concentration Response 1 Response 2 gel
temperature (C) time (min) of KOH (% w/w) viscosity (cP) strength (g/cm2)
1 70.00 60.00 8.00 739.60 51.49
2 70.00 60.00 4.64 738.30 27.41
3 60.00 30.00 6.00 746.20 26.22
4 70.00 60.00 8.00 739.60 51.49
5 60.00 90.00 6.00 1261.00 10.50
6 60.00 30.00 10.00 890.20 30.68
7 70.00 60.00 8.00 739.60 51.49
8 80.00 30.00 10.00 1291.84 94.29
9 60.00 90.00 10.00 467.50 23.88
10 70.00 60.00 11.36 741.00 75.57
11 70.00 60.00 8.00 739.60 51.49
12 70.00 9.55 8.00 934.90 49.65
13 80.00 90.00 6.00 453.00 61.57
14 80.00 30.00 6.00 494.80 50.40
15 86.82 60.00 8.00 568.60 99.71
16 53.18 60.00 8.00 910.70 3.27
17 70.00 60.00 8.00 739.60 51.49
18 70.00 110.00 8.00 544.40 53.33
19 70.00 60.00 8.00 739.60 51.49
20 80.00 90.00 10.00 312.10 114.40
6 A. Bono et al.

Figure 1 Effect of cooking temperature (C) and time (min) upon gel viscosity (cP).

but increased with the cooking temperature when % KOH was
left to be constant at 8% w/w.
Fig. 2 depicted the effect of KOH concentration and cook-
ing time and observed that at lower cooking temperature, the
gel viscosity decreased with the increasing KOH concentra-
tion, whereas the opposite trend showed at higher temperature.
Fig. 3 showed gel viscosity increased with cooking time at
lower KOH concentration, whereas at higher KOH concentra-
tion, the gel viscosity decreased.
All the interactions of between factors (cooking tempera-
ture and cooking time, KOH concentration and cooking tem-
perature, KOH concentration and cooking time) and gel

Figure 2 Effect of cooking temperature (C) and KOH concentration (% w/w) upon gel viscosity (cP).
Effect of process conditions on the gel viscosity 7

Figure 3 Effect of cooking time (min) and KOH concentration (% w/w) upon gel viscosity (cP).

viscosity can be interpreted from Eq. (1). The influence of this 3.1.2. Effect of temperature, time and KOH concentration upon
interaction is quite significant. These also can be observed gel strength of SRC
from Figs. 1–3 where the gel viscosity changes in between high- Fig. 4 showed the gel strength increased with low cooking tem-
er and lower value factors. perature on high cooking time. But the gel strength decreased

Figure 4 Effect of cooking temperature (C) and time (min) upon gel strength (g/cm2).
8 A. Bono et al.

Figure 5 Effect of concentration of KOH (% w/w) and cooking temperature (C) upon gel strength (g/cm2).

Figure 6 Effect of KOH concentration (% w/w) and cooking time (min) upon gel strength (g/cm2).

Table 3 Optimum process conditions for alkaline treatment.

Cooking temperature (C) Cooking time (min) Concentration of KOH (% w/w) Gel viscosity (cP) Gel strength (g/cm2)
80.00 30.00 10.00 1291.84 94.29
Effect of process conditions on the gel viscosity
with the cooking time at low cooking temperature and in-
creased with the cooking time at high cooking temperature.
The relationship between cooking temperature and KOH
concentration on gel strength of carrageenan is shown in
Fig. 5. From Fig. 5, it could be seen that the gel strength
increased with cooking temperature at low and high KOH
concentration. The gel strength increased with KOH concen-
tration at low and high temperature.
Fig. 6 depicted that the effect of cooking time on gel
strength is slightly changed with the variation of KOH concen-
tration. However, the strength increased significantly with the
KOH concentration at low and high cooking time.
All the interactions of between factors (cooking time and cook-
ing temperature, KOH concentration and cooking temperature,
KOH concentration and cooking time) and gel strength can be
interpreted from Eq. (2). The influence of this interaction is quite
significant. These also can be observed from Figs. 4–6 where the
gel strength changes in between higher and lower value factors.
3.2. Optimum process parameters of alkaline treatment
The result of optimization showed that the best conditions for
alkaline treatment would be as follows as displayed in Table 3.
These values were found to be related as followed by the unre-
ported local (Sabah, Malaysia) carrageenan industry.
4. Conclusion
Alkali treatment process conditions such as cooking tempera-
ture, cooking time and KOH concentration are shown as
important parameters for the production of semi refined
carrageenan.
The optimization of process parameters of alkaline treatment
was developed in this study based on the characterization of car-
rageenan; gel strength and viscosity. Considering gel viscosity
and gel strength as the main criteria to produce the best quality
of SRC and comparing with the industrial process parameters
and previous research works, the best conditions for alkaline
treatment would be as follows: cooking temperature 80 C, cook-
ing time 30 min and KOH concentration 10 (% w/w).
It could be suggested that an experimental design using the pro-
cedure called Design-Expert Software version 7 was able to deter-
mine optimum conditions for the targeted parameters of SRC.
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