TJPS Vol.

42 (Supplement Issue) 2018

Thai Journal of Pharmaceutical Sciences (TJPS)

34th International Annual Meeting in Pharmaceutical Sciences and
2nd CU FPhS - RIKEN CDB Symposium
(IAMPS34 and 2nd CU FPhS - RIKEN CDB)

Effect of pH and non-ionic polymer on in situ gelation from Gellan gum

Rawi Tishyadhigama, Angkana Tantituvanont, Jutarat Kitsongsermthon*

Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmaceutical Sciences,

Chulalongkorn University, Bangkok 10330, Thailand

* Corresponding author: Tel. +66(0)22188402; Fax. +66(0)22188401; E-mail address: Jutarat.Kit@gmail.com

Keywords: gellan gum, in situ gel, hydroxyethyl cellulose (HEC)

Introduction
The last decade, various in situ gel-forming systems have been developed and investigated.
This interest has been sparked by the advantages, such as ease of administration, reduced frequency of
administration, and improvement of patient comfort and compliance.1 In situ gel is the system which provides
rapid sol-to-gel transformation due to the existence of stimuli. There are many polymers that are used for in
situ gels such as gellan gum, alginate, pectin and poloxamer.2 Each of them needs specific stimulus, such as
pH, temperature, ions, etc, for sol-to-gel transformation.
Gellan gum is bacterial polysaccharide produced by Pseudomonas elodea. In aqueous solution at high
temperature, gellan gum is in form of a random coil, while cooling of the solution promotes the formation
of a three-fold left-handed double helix, stabilized by internal hydrogen bonding, as weak gel.3-5 Addition of
cations can reduce the effect of electrostatic repulsions among helices, augmenting the development of
stronger network. In addition, ionic bridges among carboxylic groups of neighbouring chains have been
reported to occur when divalent cations such as calcium chloride, magnesium chloride are used.8 Gellan gum
can be used alone or combining with other polymers to produce a variety of textures and there are many
reports of utilization of gellan gum for drug delivery.
The objective of this study was to investigate the effect of pH and non-ionic polymer on the gelation of
in situ gel from gellan gum. Appearance of in situ gel, viscosity, and gel capacity of formulations at different pH
(5, 6, 7) or different concentrations of hydroxyethyl cellulose (HEC) (0%, 0.15%, 0.30%) were examined.

Methods
In situ gel preparations
Six formulations were composed of ingredients as listed in Table 1. A solution of distillated water and
gellan gum were heated to 70 °C and stirred. For IG-5 and IG-6, HEC was added and mixed well with the
heated solution. When the mixture was cooled down to 40 °C, calcium chloride solution was added to obtained
0.1% w/v in the final solution. pH was adjusted with diluted sodium hydroxide or diluted hydrochloric acid for
IG-1, IG-2, and IG-3. All of formulations were stored for 24 hours at cool place.

Table 1. Formulation of in situ gel

Formulation Gellan gum HEC(%w/v) Calcium chloride Distillated Adjust pH to
code (%w/v) (%w/v) water
IG-1 0.15 - 0.1 q.s. to 100 5
IG-2 0.15 - 0.1 q.s. to 100 6
IG-3 0.15 - 0.1 q.s. to 100 7
IG-4 0.15 - 0.1 q.s. to 100 -
IG-5 0.15 0.15 0.1 q.s. to 100 -
IG-6 0.15 0.30 0.1 q.s. to 100 -

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Viscosity
The viscosity measurement was carried out using Brookfield viscometer model DVII+. The developed
formulations were placed in the beaker and spindle no. 61 and no. 63 were used for solution (without calcium
chloride) and for in situ gel (with calcium chloride), respectively, at 140 rpm.
Gelling capacity
The gelling capacity of prepared formulations was determined by placing a drop of gellan gum
solutions without calcium chloride in a beaker containing 50 ml of freshly prepared 1% w/v calcium chloride
solution. Gel formation was observed and the time for gelation and the time taken for the formed gel to
redissolve were recorded.

Result
Viscosity
The viscosity of formulations at pH 5, 6 and 7 (IG-1, IG-2 and IG-3, respectively) before adding calcium
chloride were lower than the viscosity after adding calcium chloride and this showed that the gelation occurred
in the presence of calcium ions. The viscosity of formulations at pH 5, 6 and 7 after gelation were significantly
different among each other (p-value < 0.05) and the formulation at pH 6 exhibited the highest viscosity, as
demonstrated in figure 1. Figure 2 shows the effect of HEC concentration on gellan gum gelation. The viscosity
of formulations before gelation significantly increased (p-value < 0.05) when the concentration of HEC
increased, but, after gelation, the significant differences of the viscosity was found only on the formulation with
0.30%w/v HEC (IG-6) (p-value < 0.05).

*
* *

Figure 1. Effect of pH before and after adding calcium chloride *significant difference (p-value < 0.05)

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*
* *

Figure 2. effect of HEC concentration before and after adding calcium chloride *significant difference (p-value < 0.05)

Gelling capacity
Table 2 shows the gelation time and the stability of gel of all formulations. All of formulations exhibited
rapid sol-to-gel transformation and the gelation was occurred within 30 seconds. Gels from the acidic
formulations (IG-1 and IG-2) were stable than that from the neutral formulation and they were more stable
when HEC concentration in the formulations increased. Figure 3 shows the appearance of in situ gel that
immediately occurred upon contacting with 1% w/v calcium chloride solution. Water-soluble color was added in
the gellan gum solutions for better observation. Only formulation at pH7 (IG-3) formed unshaped gel.
The formulations that contained HEC (IG-5 and IG-6) formed hard gel and the gel stability depended on
concentration of HEC.

Table 2. Gelling capacity

Formulation pH %HEC Gelation time Gel stability
IG-1 5 - +++ ++
IG-2 6 - +++ ++
IG-3 7 - +++ +
IG-4 6 - +++ ++
IG-5 5.96 0.15 +++ ++
IG-6 6.08 0.30 +++ +++
Gelation time: +++ immediately within 30 seconds
Gel stability (Gel redissolving time): + within 60 -120 minutes, ++ within 120 -150 minutes, +++ within 180 -210 minutes

HEC HEC HEC

pH 5 pH 6 pH 7 0% 0.15% 0.30%

Side view

Top view

Figure 3. Appearance of in situ gels

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Discussion
All of formulations demonstrated rapid sol-to-gel transformation by adding calcium chloride.
The viscosity of gellan gum solution decreased as the pH increased, due to the electrostatic repulsion of
ionized carboxyl groups (COO-) between the chains of gellan gum.Thus, when the pH increased, the number of
ionized carboxyl groups increased and all of them were dissociated to ionized form at pH 7 and above.2,6 When
calcium chloride was added for in situ gelation, cations could reduce the electrostatic repulsion and divalent
cations could strengthen the gel network by bonding with two chains of carboxyl groups.2,7,8 The more the
ionized carboxyl groups (by increase of pH), the stronger the gel network, as shown by higher viscosity of the
gel at pH 6 than the viscosity of the gel at pH 5. This phenomenon was not observed for the gel at pH 7
which was adjusted to pH 7 by using sodium hydroxide. For the gel at pH 7, sodium ions were already in the
solution, so sodium ions formed the ionic bond with carboxyl groups at better efficient rate than calcium.
However, sodium ion had only one valence electron, it formed a single bond with only one carboxyl group on
gellan gum chain (no interconnecting-chain network) and it resulted in forming weak gel.
HEC is non-ionic water soluble polymer with various functions, such as thickener and binder, widely
used by the cosmetic and pharmaceutical industries.9 The viscosity of solutions dramatically increased when
the concentration of HEC increased, due to its viscosity-enhancing property. Nevertheless, after gelation,
the concentration of HEC had little effect on the gel, since calcium ions induced much stronger gel network.
Therefore, the development of in situ gelling system by using HEC for adjusting the viscosity of solutions,
as in formulations, had less influence on the quality of occurred gel.

Conclusion
This study demonstrated that the gelation of gellan gum depended on pH since pH involved in the
dissociation of carboxyl groups. The optimum pH of gellan gum gelation was 6 and it was the native pH of
gellan gum. HEC, usually used as a thickener, could increase the viscosity of formulations in both solutions and
in situ gels. This combination of gellan gum and HEC can be further developed for in situ gelling drug delivery
systems.

Acknowledgements
The authors would like to thank Winner Group Enterprise PLC. for providing gellan gum
(KELCOGEL®)as gift sample and Research and Development manager of S.C. Artistry Co.,Ltd. for provision of
equipment.

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