CHARACTERISTICS OF HYDROGEL FORMULATED FROM MICRO PARTICLE OF ι-

AND κ-CARRAGEENAN COMBINED WITH P0LYVINYL ALCOHOL

ABSTRACT
Eucheuma seaweed from tropical waters habitat mainly contains carrageenan, a type of hydrocolloid
potential for hydrogel production. The aim of this study is to determine the characteristics of hydrogel
formulated from micro particle of ι- and κ-carrageenan combined with polyvinyl alcohol (PVA) which could
be used further for wound dressing application. The concentrations of ι- and κ-carrageenan used were
1.0%, 1.5%, 2.0%, 2.5% and 3.0% w/w, and the PVA concentration was 12.5% w/w. The hydrogel was
irradiated using 60Co γ-rays with the irradiation dose of 25, and 2 kGy.h-1. Concentration of ι- and κ-
carrageenan micro particle influenced the gel fraction, water absorption, tensile strength, elongation,
surface morphology and chemical structure of the hydrogel. The optimum characteristic of composite
hydrogel was obtained from 3.0% w/w of ι- and κ-carrageenan micro particle which had a gel fraction of
61.67%, water absorption 1067.12%, tensile strength 32.37×103 MPa, and elongation 251.67%.
Interestingly, the product with the optimum characteristics had a porous surface morphology,
transparent, flexible, strong as well as having high elasticity and water absorption which are applicable
for wound dressing purposes.
Keywords: Eucheuma, ι- and κ-carrageenan, micro particle, hydrogel. Commented [ASD3]: Max number of keywords is 5.

Introduction

Eucheuma seaweed, as a source 3 types of carrageenan: iota (ι), kappa (κ), and lambda (λ)

carrageenan, can be used as raw material for various applications including hydrogels (Abad

et al., 2014), but only ι- and κ-carrageenan are able to be used for hydrogel production (Li et Commented [ASD4]: In-text citations throughout the whole
document are not written in accordance to the 6th APA guidelines.
al., 2014). The synthesis of hydrogel involves heating and cooling carrageenan solution, Please use a reference manager.

freeze-thaw process and polymerization of the solution under radiation (Hoffman, 2012).

Hydrogel can be applied in chemical, biomedical or pharmaceutical products including

wound dressing. Synthesis of hydrogel for wound dressing can be combined with some other

matrices such as polyvinyl alcohol (PVA), polyvinyl pyrrolidone (PVP), and hydroxyethyl

starch. Composite of κ-carrageenan hydrogels with PVA has been reported by Kartika et al.,

(2015) and Varshney, (2007), with PVP by Abad et al. (2014) and Erizal (2008), and

combination of κ- and ι-carrageenan with PVP by Utomo et al. (2015). Most of these

experiment were using macro particle, while synthesis of hydrogels using micro particle is still

rare.

The aim of this study is to find out the optimum concentrations of ι- and κ-carrageenan

micro particles in combination with PVA to produce hydrogel. Parameters used to see the Commented [ASD5]: These aims are different from those in the
abstract, which one is correct?
quality of the hydrogel include gel fraction, water absorption, tensile strength, elongation, and

surface morphology.

Material and methods Commented [ASD6]: Mention all chemicals used in this study,
including the grades and the brands of each chemical.

1
Materials used in this study were Eucheuma denticulatum and Kappaphycus alvarezii

seaweeds harvested from Nusa Penida Island, Bali, Indonesia. Quality of the dry seaweeds Commented [ASD7]: Month and year of collection? How were
they preserved? Were they collected as dry or fresh seaweed?
as raw material were evaluated by analyzing their moisture content, clean anhydrous weed

(CAW) and impurities based on Indonesian National Standard (SNI) method of analysis for dry

seaweeds (Badan Standarisasi Nasional, 2015).

Extraction of ι- and κ-carrageenan

Extraction process for E. denticulatum was conducted as follows: 2000 g of dry seaweed

was soaked in water for 30 min, washed and rinsed several times. The seaweed was treated

with 12 L of 8% KOH at 60-65 °C for 120 min, rinsed with water and extracted using 40 L of

water at 80-85 °C for 120 min. Celite (3% w/w) was added in the extraction process. The

extract was mixed with 99% isopropyl alcohol (IPA) with filtrate to IPA ratio of 1: 2 (v/v) with

continuous stirring untill ι-carrageenan fibre was formed. The carrageenan fiber was dried in Commented [ASD9]: What temperature?

the sun. Extraction process for K. alvarezii was conducted with similar procedure as E.

denticulatum until filtrate was produced. The filtrate was cooled down to a temperature of 25

°C so that κ-carrageenan gel was formed. The gel was put in the filter cloth, pressed and then

sun dried.

Preparation of micro particle

ι-carrageenan produced was firstly milled using hammer mill and subsequently milled using

ultra centrifugal mill (UCM) ZM 200 Retsch with 0.08 mm sieve and 6000 rpm speed, and then

filtered using Vibratory Sieve Shaker (VSS) AS 200 Retsch of 200 mesh size.

Characterization of ι- and κ-carrageenan micro particle

To see the chemical bonds of the carrageenan compound, analysis was conducted using

Fourier transform infrared spectrophotometer (FTIR) Perkin Elmer. The samples were coated Commented [ASD10]: Series?

with potassium bromide (KBr) using a vacuum pump and then analyzed using FTIR at a

wavenumber between 4000-450 cm-1. In order to measure the particle size, samples were

analyzed using particle size analyzer (PSA) (Beckman, Delsamax Pro, Coulter). Samples were Commented [ASD11]: Series?

firstly dispersed using alcohol, filled into an ultrasonic bath for 10 min and then analyzed. The

2
surface morphology of micro particles was then analyzed using scanning electron microscope

(SEM) (JEOL, type, country origin). Samples of ι- and κ-carrageenan micro particles was firstly Commented [ASD12]: This has not been addressed.

coated with gold powder in a smart coater and then observed under SEM with 500 times

magnification.

Preparation of composite hydrogel

PVA solution (100 mL; 12,5% w/v) was autoclaved for 15 min, and blended with 100 mL

mixture of κ- and ι-carrageenan micro particle in a proportion of 6 : 4 by weight (Utomo et al.,

2015). Various concentrations ie. 1.0%; 1,5%; 2.0%; 2.5%; and 3.0% of the mixture were

prepared and heated at 80 oC for 15 min with continuous stirring until homogenous solution

was formed (Dafader et al., 2015; Erizal, 2008; Varshney, 2007). The solution was poured into

plastic containers (30 x 16.5 x 0.3 cm3) and cooled down to 25 °C for about one hour until

hydrogel was formed. The hydrogel was then dehydrated using freeze-thaw process. The

freezing temperature was -5 °C conducted in a freezer for 18 hours, and thawing was done for

about 8 hours. The process was conducted in a 4 cycles of freeze-thaw process (Kartika et

al., 2015; Kenawy et al., 2014). The hydrogel were irradiated with γ rays at the dosage of 25
60
kGy (dosage rate of 2 kGy/h) using Co irradiator IRKA PATIR BATAN (Brant et al., 2012;

Dafader et al., 2015; Varshney, 2007).

Characterization of Composite Hydrogel

Gel fraction and water holding capacity

Gel fraction and water holding capacity (WHC) tests were conducted using method of Dafader

et al. (2015); Kenawy et al. (2014); and Varshney (2007). Hydrogel sample was cut into small

pieces of 2×2×0.5 cm3, dried in an oven at 60 C for 24 h. For the gel fraction test, the dry

samples were packed in a stainless steel screen of 300 mesh. The screen containing sample

was then soaked in distilled water at a temperature of 80 C in a waterbath shaker set at 100

rpm for 24 h to remove dirth. Samples were then taken from the waterbath and re-dried in an
o
oven at 60 C until a constant weight was achieved. For WHC test, dry samples were soaked

in distilled water for 24 h and then weighted (A g). The sample was then dried in an oven at a
o
temperature of 60 C for 24 h (B g).

3
 Weight of dry clean sample (g)
Gel fraction = --------------------------------------------- x 100%
Weight of dry original sample (g)

(A – B) g
WHC = ------------------- x 100%
Bg

Tensile strength and Elongation test

Tensile strength test of hydrogel was conducted using strograph-R1 Toyeseki (Erizal, 2008;

Varshney, 2007; Wu et al., 2001). Sample with the dimension of 10×0.6×0.2 cm3 was cut into

a dumbbell shape to get a standard size and shape of ASTM standard. Sample was tested Commented [ASD15]: Spell out at first mention.

using strograph-R1 for tensile strength and elongation test.

SEM Analysis

Hydrogel sample was characterized using SEM to see the surface morphology. The sample

was firstly freeze-dried for 2×24 h then coated with gold in a smart coater. The surface

morphology was analyzed using SEM at 500 magnifications.

FTIR Analysis

To see the functional group of the hydrogel, sample was analyzed using FTIR at wavenumber

between 4000-450 cm-1. The sample was firstly freeze-dried for 2×24 h then coated with KBr

powder using specac in vacuum condition. Commented [ASD16]: Replace these paragraphs with “SEM and
FTIR analyses were conducted as previously described” to avoid
redundancy.

Results and Discussion

The quality of raw materials

Moisture content of the dry K. alvarezii and E. denticulatum were 36.66% and 38.97%,

respectively. These results were higher than national standard (SNI 2690:2015) in which

maximum moisture contents must be 30.0%. Clean anhydrous weed (CAW) values of K. Commented [ASD17]: Only spelled out at first mention, use
abbreviations afterwards.
alvarezii and E. denticulatum were 49.50% and 42.94%, respectively. These values were

within SNI 2690:2015 of max 50.0%. In the previous research (Utomo et al., 2015) the CAW

of K. alvarezii and E. denticulatum were 49.50% and 54.67%, respectively. The slightly low

CAW values of K. alvarezii and E. denticulatum were resulted from the high content of dirth,

sand, salt, and rock. The impurities of K. alvarezii was 4.65% higher than the standard of SNI

2690:2015 i.e. below 3.0% and E. denticulatum was 1.85% which was well within the standard

4
of SNI 2690:2015. However, the impuritis of K. alvarezii was higher than the SNI standard.

The high impurity of K. Alvarezii was probably caused by the high content of mud since this

seaweed was cultivated not very far from the estuary.

Micro particle of ι- and κ- carrageenan

The dried carrageenan both from E. denticulatum and K. alvarezii were milled and filtered

producing micro particle powder of ι- and κ-carrageenan. Carrageenan in micro particle form

has a better physico-chemical character than macro particle. Physical treatment through

milling was very effective to produce micro particle size of carrageenan powder. Commented [ASD20]: I suggest that these paragraphs are
removed since they do not contain results from the current study.
In this study, the particle size of ι- and κ-carrageenan were measured both using PSA.

Based on PSA results (Fig. 1), from 10 samples measured, shown by different colours, the

mean diameter of ι-carrageenan (Fig. 1a) was 686 nm or 0.68 μm and that of κ-carrageenan

(Fig 1b) was 641 nm or 0.64 μm. The intensity of of ι-carrageenan is higher than that of κ-

carrageenan, in contrast, the distribution of the radius is wider κ-carrageenan than that of ι-

carrageenan. This means that the particles size of ι-carrageenan is more homogenous than

that of κ-carrageenan.

a b

Commented [BR21]: Elaborate the caption here, about what are

Fig. 1 Micro particle size of carrageenan measured using PSA (a) ι- and (b) κ-carrageenan. the several different color line mean?
From 10 samples measured, shown by different colours, the mean diameter of ι-carrageenan
Commented [ASD22R21]: This has not been addressed. What
(a) was 686 nm or 0.68 μm and that of κ-carrageenan was 641 nm or 0.64 μm does the color blue, pink, yellow etc. indicate? Put the explanation in
the caption.

5
Chemical Structure

FTIR analysis is conducted to look at functional group of ι- dan κ-carrageenan micro particle.

FTIR Spectrum of ι- and κ-carrageenan micro particle is shown in Fig. 2 and the wavenumber

data of the spectra is presented in Table 1. Both spectra has a wavenumber of 1262 cm-1

showing that micro particle ι- and κ- carrageenan has S=O group from sulphate ester.

Glicosidic bond of β(1-4) on κ- and ι-carrageenan micro particle were shown by wave number

of 1042 and 1028 cm-1. Wave number of 928 cm-1 on κ-carrageenan and 933 cm-1 on ι-

carrageenan micro particle shows C–O group of 3,6-anhydro-D-galactose. Wave number of

846 cm-1 and 850 cm-1 shows C–O–S group of D-galactose-4-sulphate of κ- and ι-carrageenan

micro particle. These results is in accordance with the study conducted by Relleve et al. (2015)

and Duan et al. (2016).

Fig. 2 FTIR spectra of (a) ι- and (b) κ- carrageenan micro particle

Table 1 Characteristics of infrared absorbtion of κ- and ι-carrageenan micro particle

Wavenumber (cm-1) Functional group

κ-carrageenan ι-carrageenan
- 806 C–O–S of 3,6-anhydro-D-galactose-2-sulphate
846 850 C–O–S of D-galactose-4-sulphate
928 933 C–O stretching of 3,6-anhydro-D-galactose
1042 1028 C–O–C stretching of glycosidic β(1-4)
1262 1262 O=S=O stretching of Ester sulphate

6
 The difference between ι- and κ-carrageenan micro particle is shown by the absorbtion

at wave number of 806 cm-1 which shows C–O–S group of 3,6-anhydro-D-galactose-2-

sulphate of ι-carrageenan. Wave number between 800-850 cm−1 shows 3,6-anhydro-D-

galactose-2-sulphate and D-galactose-4-sulphate. Wave number area of 800-850 cm−1 shows

the sulphate group of carrageenan (Relleve et al., 2015). Based on this analysis, ι- and κ-

carrageenan micro particle produced from the extraction are actually ι- and κ-carrageenan.

Characteristics of composite hydrogel Commented [ASD23]: Mention the standard characteristics, if

applicable.
Visual and organoleptic characteristics of the hydrogel prepared from micro particle of ι- and

κ-carrageenan at various concentrations can be seen in Fig 3. The hydrogel was transparent,

flexible, strong and elastic.

Fig. 3 The typical hydrogel produced from this experiment with ι- and κ-carrageenan
concentrations of (a) 1%, (b) 1,5%, (c) 2%, (d) 2,5%, (e) 3% w/w loadings, it was
transparent, flexible, strong, elastic and having a high water holding capacity

Results of physical characteristics analysis of hydrogel in various concentration ι- and

κ-carrageenan micro particle covering gel fraction and water holding capacity using Commented [ASD24]: Abbreviate.

gravimetry, as well as tensile strength and elongation measured using Toyeseki instrument

are presented in Fig 4.

Gel fraction of the composite tended to decrease with increasing carrageenan

concentration, Fig. 4a shows that the gel fraction was 80.91% when the carrageenan

concentration was 1% until finaly the gel fraction became 61.67 when the carrageenan

concentration was 3%. This is unusual fenomena since the addition of carrageenan should

increase the gel fraction of the composite since carrageenan itself is a gel forming substance.

It was probably that PVA had more profound effects than carrageenan, with higher

concentration of carrageenan indirectly lower the concentration of PVA that this will lower the

7
gel fraction of the composite. Besides, there are some factors affecting gel fraction of the

hydrogel such as polymer composition and irradiation dose. The gel fraction ranged between Commented [ASD25]: Please provide references to support this
argument.
61.67-80.91%. The highest gel fraction is owned by hydrogel product with the lowest
Commented [ASD26]: Is there a range of standard values or
concentration of carrageenan. The high concentration of hydrogel concentration could have those of commercial ones that you can compare these numbers to?

the high proportion of cross linking resulted from γ-ray irradiation process so that PVA react

with ι- and κ-carrageenan micro particles. Hydrogel with concentration of κ- and ι-carrageenan Commented [ASD27]: Please provide references to support this
argument.
micro particle 3% had the lowest gel fraction i.e. 61.67%. The increase of concentration κ- and

ι-carrageenan micro particle up to 3% could possibly cause degradation of κ- and ι-

carrageenan micro particle caused by γ-ray irradiation process. Commented [ASD28]: Please provide references to support this
argument.

85 a 1200
80 b
Gel Fraction (%)

75 1000
WHC (%)

70
800
65
60 600
55
50 400
1 1.5 2 2.5 3 1 1.5 2 2.5 3
i- and K- carrageenan (%)
i- and K- carrageenan (%)

40 300
c d
Tensile strength (MPa)

250
30
Elongation (%)

200
20 150
100
10
50
0 0
1 1.5 2 2.5 3 1 1.5 2 2.5 3
i- and K- carrageenan (%) i- and K- carrageenan (%)

Fig. 4 Correlation between concentration and physical characteristics of carrageenan (a) gel
fraction, (b) water holding capacity (WHC), (c) tensile strength (d) elongation

Different from the gel fraction fenomena, the increasing concentration of carrageenan,

the water holding capacity of the composite was also increased. The water holding capacity

started from 531.17% when the carrageenan concentration was 1% and then continuously

increased until 1067.12 when carrageenan concentration reaching 3%. The characteristic of Commented [ASD29]: Is there a range of standard values or
those of commercial ones that you can compare these numbers to?
carrageenan is very strong in retaining water consequently the higher concentration of

carrageenan applied in the composite the higher was the the water holding capacity. The

characteristics of this composite in term of tesile strength and elongation (Fig 4c and 4d), these Commented [ASD30]: When transitioning from one topic to the
other, please use a different paragraph.

8
two parameters had similar pattern with water holding capacity in that they increased with the

increasing carrageenan concentration. When the concentration of carrageenan 1% the tensile

strength and elongation were 9,330 MPa and 113.33%, respectively, these parameters

increased gradually until the concentration of carrageenan was 3%. At this point, the tensile

strength was 32,370 Mpa and the elongation was 251.67 %. Figure 4a shows that gel fraction
decreased with increasing carrageenan concentration from 1-3%. While water holding
capacity, tensile strength, and elongation increased with increasing carrageenan
Commented [ASD31]: Is there a range of standard values or
those of commercial ones that you can compare these numbers to?
Commented [ASD32]: Focus on the figure that you are
discussing to avoid confusion. This paragraph discusses Fig 5b-d.
Please remove this sentence.

concentration. Gel fraction, water holding capacity, tensile strength and elongation of the

hydrogel were very much affected by the numbers of cross link (Brant et al., 2012).

For comparation, tensile strength and elongation of Medifoam® N has tensile strength

0.033 kgf mm−2. The mean tensility of other 8 commercially available polyurethane (PU) foam

dressing ranged about 0.01 kgf mm−2 ~ 0.025 kgf mm−2. The greatest elongation properties

among 8 dressings tested was 1101 %. Medifoam® N has also good elongation property

(412 %). The remaining 8 products tested were ranged around 180–377 %. (Lee, et al. 2016).

So far no certain quality standard for these parameters.

Based on research conducted by Varshney (2007) the optimum concentration of PVA

was 8%, Dafader et al., (2015); Kenawy et al., (2014); and Brant et al., (2012) using PVA

concentration of 10% and Wu et al., (2001) using concentration of PVA 15%, PVA

concentration of this research used was 12.5% with four cycle freeze-thaw process conducted

to synthesize the hydrogel. According to Kenawy et al., (2014), hydrogel formulated with PVA Commented [ASD33]: Separate this sentence for clarity. Please
explain why you chose PVA 12.5%.
can be synthesized using freeze-thaw process. Hydrogel which is synthesized with

PVA/PVP/PU through freeze-thaw process followed γ-ray irradiation is able to improve the

physico-chemical characteristics (gel fraction, tensile strength and elongation) compared to

those only using γ-ray irradiation (Park et al., 2003). According to Yang et al., (2008), hydrogel

from PVA/chitosan processed with γ-ray irradiation followed by freeze-thaw process had water

holding capacity, mechanical characteristics, and thermal stability higher than those only

treated with freeze-thaw process followed with γ-ray irradiation. The irradiation using dose of

25 kGy was based on a study conducted by Brant et al., (2012) and Dafader et al., (2015), this

dose is also recommended for sterilization process for health products (Varshney, 2007).

9
 The effects of irradiation to hydrogel depend on compatibility degree, the area of

molecular interaction and type of homopolymer used. Compatibility is affected by molecular

weight of the homopolymer. Among micro particle of ι- and -κ carrageenan as well as PVA

enable them to make hydrogen bonds, ionic bonds and electrostatic interaction. So that cross

linking or degradation of ι- and -κ carrageenan micro particle as well as PVA can be happen

on the hydrogel with a certain irradiation dose.

According to Dafader et al., (2015) when hydrogel synthesized from PVA and

carrageenan and exposed to irradiation with a certain dose, the carrageenan will be degraded

into lower molecular weight compounds. Commented [ASD36]: Mention your results first, then compare
them to the others.
The degraded carrageenan molecules are able to prevent the cross linking formation Commented [ASD37]: How much would be degraded? Can you
measure it? Or can you estimate?
between PVA and carrageenan resulting the decrease of the gel fraction together with the

increase of carrageenan concentration. The addition of ι- and κ-carrageenan micro particle

from 1 to 3% on the hydrogel can decrease gel fraction produced. On the other hand, water Commented [ASD38]: This paragraph should be move in the
paragraph discussing gel fraction.
holding capacity, tensile strength, and elongation increase with increasing concentration of ι-

and κ-carrageenan micro particle up to 3%.

Water holding capacity is a parameter showing the capacity of the hydrogel to hold

water and swell. As stated by Carabeo (2005), a substance with water holding capacity more

than 10% from its initial weight is called hydrogel, and those with water holding capacity more

than 95% from its initial weight is called super absorben hydrogel. Fig. 5(b) shows that the

WHC ranged from 531.17 to 1067.12%, the higher concentration of the hydrogel the higher is

its WHC since the hydrophilic group of the hydrogel is higher. When hydrophilic group of

hydrogel was ionised, its WHC increases due to the higher electrostatic force (Dafader et al.,

2015). These phenomena showed the interaction between ι- and κ- carrageenan micro particle

with PVA, water holding capacity increases proportionally with the increase of ι- and κ-

carrageenan concentration. Commented [ASD39]: This paragraph should be move upward.

We have passed the discussion of 5c and d. Here we are going back
According to Varshney (2007), mechanical properties of hydrogel can be represented to 5b. Please rearrange this paragraph for flow and clarity.

by tensile strength and elongation which are closely related to flexibility. It can be seen from

Fig. 5(c), the tensile strength of the hydrogel ranged between 9.33×103 to 32.37×103 MPa.

The highest tensile strength is achieved by hydrogel with highest concentration of ι- and κ-

carrageenan micro particle. It could be due to the higher cross linking between ι- and κ-

10
carrageenan micro particle with PVA. According to Kenawy et al. (2014), polysaccaride

addition can increase tensile strength since the the cross link is also increased. Tensil strength

of PVA hydrogel added with polysaccaride (such as carrageenan) can directly increase tensile

strength and elongation (Varshney, 2007). This indicates that there is a formation of three

dimensional structure of the hydrogel. The addition of ι- and κ-carrageenan micro particle

concentration from 1 to 3% of hydrogel was able to increase tensile strength of the hydrogel.

In contrast, elongation is closely related to elasticity of hydrogel (Varshney, 2007). Fig. 5(d)

shows the elongation ranged between 113.33-251.67%. The highest elongation is achieved

by hydrogel with the highest concentration of ι- and κ-carrageenan of 3% it might be caused

by the higher proportion of cross link between ι- and κ-carrageenan with PVA (Sabangan,

2008). A study conducted by Rosiak et al. (2001) found that the addition of polysaccaride can

increase elongation, however, too much polysaccaride will decrease hydrogel elongation

(Kenawy et al., 2014). This is in accordance with Sabangan (2008), that if some PVA is

hydrolyzed the elongation will increase. Commented [ASD41]: Rearrange the position of this paragraph.

Surface morphology of the hydrogel was observed under Scanning electrone

microscope (SEM). This method has been used by several researchers (Dafader et al., 2015;

Hezaveh et al., 2013a; Kenawy et al., 2014; Varshney, 2007). According to Nanaki et al.,

(2015), SEM can be used to see not only surface morphology but also physico-chemical

characteristics of hydrogel. Results of SEM analysis of ι- and κ-carrageenan micro particle

with 500 magnifications can be seen in Fig. 5. Commented [ASD43]: Mention your results first. Then compare
them with those of other studies.

a b

c d e

Fig. 5 Micrograph of hydrogel surface of ι- and κ-carrageenan at concentrations of (a) 1%,

(b) 1,5%, (c) 2%, (d) 2,5%, (e) 3% w/w loadings

11
 Fig. 5 shows that surface morphology of ι- and κ-carrageenan microparticles have

porous surface with various size. Inner part of freeze-dried hydrogel shows many porous

structures The porous texture of hydrogel was probably caused by evaporation of water

contained in the hydrogel at the time of freeze-dry process. According to Varsney (2007),

hydrogel with porous texture can absorp water and the unlimited expansion until the hydrogel

become fragile and broken down.

Spektrum FTIR micro particle of ι- and κ-carrageenan, PVA and hydrogel is presented

in Fig. 6.

Fig. 6 FTIR Spectra (a) κ-carrageenan, (b) ι-carrageenan, (c) PVA, and (d) 1% hydrogel, (e)
1,5% hydrogel, (f) 2% hydrogel, (g) 2,5% hydrogel, (h) 3% hydrogel
Functional group characteristics of hydrogel wound dressing are listed in Table 2.
Absorbtion on wavenumber 1260-1262 cm-1 on the hydrogel shows that there is a sulphte
Commented [ASD44]: This figure is redundant with Fig.3 (it
contains similar information for the carrageenans). The numbering in
the figure is not in order. The spectra is overlapping and difficult to
read. Please indicate with a straight line, the diagnostic wavenumbers
of k- and i-carrageenans, and PVA.
Commented [ASD45]: When discussing wavenumbers, start
from the highest number to the lowest. Do not go back and forth to
avoid confusion.

ester group of O=S=O. Absorbtion on wavenumber 923-927 cm-1 shows group of C–O of 3,6- Commented [BR46]: Full stop

anhydro-D-galactose. Absorbtion on wavenumber 850-853 cm -1

is C–O–S group of D- A sentence consist of one subject, one verb, if you have two subject
OR one subhect two verb so that is not correct.

galactose-4-sulphte. Glycosidic bound of β(1-4) hydrogel is seen at absorbtion on

wavenumber 1093-1097 cm-1. Absorbtion on wavenumber 3391 and 2943 cm-1 on PVA shows Commented [ASD47]: These sentences are redundant. They
have been mentioned before. Please remove.
the group of O–H and C–H, while absorbtion on wavenumber 3650-3590 cm-1 shows the group Commented [ASD48]: Are these overlapping with those from
carrageenans?
-1
of O–H free alcohol (Kenawy et al., 2014). Absorbtion on wavenumber 1096 and 919 cm on Commented [ASD49]: Are these overlapping with those from
carrageenans?
-1
PVA identify C–O group and O–H. Absorbtion on wavenumber 1144 cm on PVA shows C–

12
O–H group. Absorbtion on wavenumber 1100 and 927 cm-1 are C–O of secondary alcohol and

O–H on PVA (Khan et al., 2014).

Table 2 Functionl group chrcteristics of hydrogel wound dressing

Wavenumber (cm-1) Functionl group
ι-carrageenan κ-carrageenana PVA Hydrogel Commented [ASD50]: Unfortunately, the diagnostic
806 - - - C–O–S wavenumbers for hydrogel are not apparent from the Figure
(absorbance appearance is too low). Maybe separate the spectra of
850 846 - 853-850 C–O–S
samples from those of the carrageenans and PVA.
933 928 - 923-927 C–O stretching
- - 919 - O–H bending
1028 1042 - - C–O–C stretching
- - 1096 1096-1093 C–C stretching
- - 1144 - C–O–H stretching
1262 1262 - 1262-1260 O=S=O stretching
- - - 2921-2927 C–H stretching
- - 2943 - C–H stretching
2975 2961 - - C–H stretching
3466 3434 3391 3455-3435 O–H stretching

The changes of functional group on hydrogel is characteried by the missing of the

absorbtion on wavenumber 1144 nd 919 cm-1 showing the stretching of C–O–H group and

absorbtion on O–H group on PVA and absorbtion on wavenumber 806 cm-1 which shows the

absorbtion of C–O–S group of 3,6-anhydro-D-galactose-2-sulphate of ι-carrageenan micro

particle. The changes of the weak absorbtion on wavenumber 3435-3445 cm-1 shows the

decrease of O–H group of ι- and κ-carrageenan micro particle as well as PVA.

Conclusion Commented [ASD53]: Mention the best treatment.

The physical characteristics of the hydrogel produced was transparent, flexible, strong, and

elastic. Concentration of ι- and κ-carrageenan micro particle affected significantly on the water

holding capacity, tensile strength and elongation of hydrogel. Concentration of ι- and κ-

carrageenan micro particle also affected significantly on surface morphology and cross linking

process of hydrogel chemical structure. In this study, the best concentration of ι- and κ-

carrageenan micro particle in hydrogel formula was 3%, so that further research using higher

concentration of ι- and κ-carrageenan micro particle as well as optimum irradiation dose needs

to be conducted to get a better result in conjuntion with the application of hydrogel for woud

dressing.

13
Acknowledgments Commented [ASD54]: Add this section and mention the funding
sources of this study (grant number?)
The authors would like to thank to the Ministry of Marine Affairs and Fisferies of Republic of

Indonesia, for funding this research.

References Commented [ASD55]: References are not written in accordance

to the 6th APA guidelines. Please use a reference manager.
Abad, L. V., Aranilla, C. T., Relleve, L. S., & Rosa, A. M. D. (2014) Emerging Applications of
Radiation-Modified Carrageenans. Nucl Inst Meth Phys Res B. 336:167-172.
http://doi.org/10.1016/j.nimb. 2014.07.005.
Brant, A. J. C., Giannini, D. R., Pessoa J. O. C. P. , Andrade, A. B., Riella, H. G., & Lugao, A.
B. (2012) Influence of Dissolution Processing of PVA Blends on The Characteristics of
Their Hydrogels Synthesized by Radiation - Part I: Gel fraction, Swelling, and
Mechanical Properties. Rad Phys Chem 81:1465-1470. http://doi.org/
10.1016/j.radphyschem.2012.01.048.
Carabeo, M. M. C. (2005) Swelling Capacity of Different Polyvinyl Alcohol and κ-Carrageenan
Blend Films. Sci Chem. BSc Thesis, Mapúa Inst Technol 34 pp
Badan Standarisasi Nasional (BSN) - Indonesian National Standardization Board. (2015)
Rumput Laut Kering-Dry Seaweed. SNI 2690: 2015
Dafader, N. C., Manir, M. S., Alam, M. F., Swapna, S. P., Akter, T., & Huq, D. (2015) Effect Of
Kappa-Carrageenan On The Properties Of Poly(Vinyl Alcohol) Hydrogel Prepared By
The Application Of Gamma Radiation. Transaction 2(1):1-12.
http://doi.org/10.15764/STAC.2015.01001.
Darwis, D., Nurlidar, F., Warastuti, Y., & Hardiningsih, L. (2010) Pengembangan Hydrogel
Berbasis Polivinil Pirolidon (PVP) Hasil Iradiasi Berkas Elektron Sebagai Plester
Penurun Demam. Sains Dan Teknologi Nuklir Indonesia (Hydrogel Development Based
on Polyvinil Pyrolidon (PVP) Produced from Electron stray as Fever Reducing Band).
Indon Nucl Sci Technol 57-66. ISSN 1411-3481.
Duan, F., Yu, Y., Liu, Tian, L., & Mou, H. (2016) An Effective Methode for the Preparation of
Carrageenan Oligoscchrides Directly from Eucheuma cottonii Using Cellulase and
Recombinant κ-carrageenase. Alga Res 15:93-99.
Erizal. (2008) Pengaruh Pembalut Hydrogel Kopolimer Polivinilpirrolidon (PVP)-κ-
Carrageenan Hasil Iradiasi dan Waktu Penyembuhan pada Reduksi Diameter Luka
Bakar Tikus Putih Wistar (Effects of Hydrogel Copolymer Polyvinilpirrolidon (PVP)-κ-
Carrageenan wound dressing from Irradiation and Healing Time on Diameter Reduction
of Wisntar Mouse Burn Wound). Indon J Chem 8(2): 271-278.
Hezaveh, H., & Muhamad, I. I. (2013) Controlled Drug Release via Minimization of Burst
Release in pH-response kappa-Carrageenan/Polyvinyl Alcohol Hydrogels. Chem Eng
Res Desn 91(3):508-519. http://doi.org/ 10.1016/j.cherd. 2012.08.014.
Hoffman, A. S. (2012) Hydrogels for Biomedical Applications. Adv Drug Deliv Rev 64:18-23.
http://doi.org/10.1016/j.addr.2012.09.010.
Kartika, R., Gardi, A., & Darma, G. C. E. (2015) Formulasi Basis Sediaan Wound dressing
Hydrogel dengan Teknik Beku Leleh Menggunakan Polimer Kappa Carrageenan
(Supply Basis Formulation for Wound Dressing using Freeze-Thaw Technique of
Kappa-Carrageenan Polymer). Unisba. 643-648. ISSN 2460-6472.
Kenawy, E., Kamoun, E. A., El-Meligy, M. A., & Mohy, M. S. (2014) Physically Crosslinked
Poly(vinyl alcohol)-hydroxyethyl Starch Blend Hydrogel Membranes : Synthesis and
Characterization for Biomedical Applications. Arab J Chem 7(3):372-380.
http://doi.org/10.1016/j.arabjc.2013.05.026.
Lee, S.M., Park, I.K., Kim, Y.S. et al. (2016) Physical, morphological, and wound healing
properties of a polyurethane foam-film dressing. Biomater Res 20, 15 (2016).
https://doi.org/10.1186/s40824-016-0063-5
Li, L., Ni, R., Shao, Y., & Mao, S. 2014 Carrageenan and Its Applications in Drug Delivery. A
Review. Carbohydr Polym. 103:1-11. http://doi.org/10.1016/j.carbpol.2013. 12.008.
Nanaki, S. G., Kyzas, G. Z., Tzereme, A., Papageorgiou, M., Kostoglou, M., Bikiaris, D. N., &
Lambropoulou, D. A. (2015) Synthesis and Characterization of Modified Carrageenan

14
 Micro particles for The Removal of Pharmaceuticals from Aqueous Solutions. Coll Surf
B: Biointerfaces. 1-36. http://doi.org/10.1016/ j.colsurfb.2015.01.053.
Park, K. R., & Nho, Y. C. (2003) Synthesis of PVA/PVP Hydrogels Having Two-Layer by
Radiation and Their Physical Properties. Rad Phys Chem. 67:361–365.
http://doi.org/10.1016/S0969-806X(03)00067-7.
Razzak, M. T., Darwis, D., Zainuddin, M., & Sukirno. (2001) Irradiation of Polyvinyl Alcohol
and Polyvinyl Pyrrolidone Blended Hydrogel for Wound Dressing. Rad Phys Chem.
62:107-113.
Relleve, L. & Abad, L. (2015) Characterization and Antioxidant Properties of Alcoholic Extracts
from Gamma Irradiated κ-Carrageenan. Rad Phys Chem.
http://doi.org/10.1016/j.radphyschem. 2015.02.028.
Rosiak, M. T., Darmawan, D., & Zainuddin, S. (2001) Irradiation of polyvinyl alcohol and
polyvinyl pyrrolidone mixed hydrogel for wound dressing. Rad Phys Chem, 62 (2001),
pp. 107-113
Sabangan, K.I. (2008) The Effect of Blend Ratio on the Mechanical Properties of Different
Polyvinyl Alcohol and Kappa - Carrageenan Blend Films. Science in Chemistry. Mapua
Institute of Technology.
Utomo, B. S. B., Peranginangin, R., Suryaningrum, D., Darmawan, M., Fransiska, D., &
Rahmad, W. (2015) Teknologi Formulasi Campuran Iota dan Kappa Carrageenan
Murni dalam Pembuatan Hydrogel. Laporan Teknis Penelitian (Formulation Technology
of Pure Iota and Kappa-Carrageenan in Hydrogel Processing. Tech Res Rep) 1-30.
Varshney, L. (2007) Role of Natural Polysaccharides in Radiation Formation of PVA–Hydrogel
Wound Dressing. Nucl Inst Meth Phys Res B. 255:343-349. http://doi.org/
10.1016/j.nimb.2006.11.101.
Wu, M., Bao, B., Yoshii, F. & Makuuchi, K. (2001) Irradiation of Crosslinked, poly(vinyl alcohol)
Blended Hydrogel for Wound Dressing. Radioanal Nuclear Chem 250(2):391-395.
Yang, X., Liu., Q., Chen, X., Yu., F., & Zhu, Z. (2008) Investigation of PVA/ws-Chitosan
Hydrogels Prepared by Combined γ-Irradiation and Freeze-Thawing. Carbohydr Polym
73:401-408. http://doi.org/10.1016/ j.carbpol.2007.12.008.

15