Devarajan Saravanan et al.

/ Journal of Pharmacy Research 2011,4(6),1633-1636

Research Article Available online through
ISSN: 0974-6943 http://jprsolutions.info
Formulation and evaluation of sustained release glucomannan-
sodium alginate beads; Isolated from Araucaria cunninghamii Linn.
Devarajan Saravanan1*, Chembeti Praveen Kumar 1 , Kolleboyina Mallikarjuna Rao 1 ,
Chembeti Pradeep Kumar 1, Yerrabrolu Surendra Reddy 1 , Tirampuram Lokesh1
1
Ratnam Institute of Pharmacy, Pidathapolur, Nellore, Andhra Pradesh-524346.
Received on: 11-02-2011; Revised on: 16-03-2011; Accepted on:21-04-2011
ABSTRACT
In the present work, glucomannan (polysaccharide used as antidiabetic and antihyperlipidemic activity) has been isolated from the seeds of Araucaria cunninghamii by a simple
method without using toxic chemicals. The glucomannan content was found to be about 5-9% (w/w) in original Araucaria seeds. The isolated glucomannan was characterized by FTIR
and Mass Spectroscopy and was used for the preparation of glucomannan-sodium alginate sustained release beads. Five formulations of glucomannan-sodium alginate sustained release
beads were prepared by using increasing concentrations of sodium alginate (1 - 5%w/v), calcium chloride (2%w/v) solution as a gelling agent and 0.5%w/v of Cunninghamii
glucomannan (CGM) as active pharmaceutical ingredient. The prepared beads were characterized by FTIR and evaluated for post formulation parameters such as swelling index, angle
of repose, tapped density, bulk density, porosity, Carr’s index and in-vitro drug release. The surface morphology of prepared beads was studied by SEM analysis. From the above
research work the results revealed that, as the concentration of sodium alginate increases, the percentage cumulative drug release has been retarded. So among the five formulation G 5
(sodium alginate (5%) and CaCl2 (2%)) was considered as a best formulation in the present research work.

Key words:Isolation, Cunninghamii glucomannan, sodium alginate, sustained release, beads, in-vitro dissolution.

INTRODUCTION
Both natural and synthetic polymeric systems have been investigated for the controlled release
of drug. Hydrophilic polyionic carbohydrates such as alginate (ALG) 1, 2 have been paid much
attention in recent years. Since the preparation of beads by these materials involves the use of
aqueous solvents, environmental problems associated with organic solvents would be minimized.
These beads would also have no immunogenicity and bio adhesive properties3, which could
serve as a potential advantage in mucosal drug delivery.ALG, a naturally occurring copolymer
of guluronic and manuronic acids, can be ionically cross-linked by the addition of divalent
cations in aqueous liquid. The relatively mild gelation process has enabled not only proteins4
but also cells5 and DNA6 to be incorporated into ALG matrices with retention of full biological
activity. ALG is chemically very stable between a pH of 5 and 10. But high acid concentrations
causes decarboxylation of ALG. Therefore in order to improve the stability of ALG capsules in
acidic gastric juice, complex coacervation of oppositely charged polyelectrolytes such as
chitosan (CHI) 7 and polylysine (PLL) has been commonly used8.
It has been recently reported that the drug delivery particulates were prepared using ALG, PLL
and pectin to release theophylline, chlorthiazide and indomethacin9. Pectin is more resistant to
cleavage in the gut than ALG gel. It breaks down in the presence of micro-flora in the colon and
has been used as colonic drug. Use of pectin especially helped in forming a more robust
particulate that was more resistant in acidic pH and modulated the release profiles of the
encapsulated model drugs in the alkaline pH. This particulate system may have potential use
as a carrier for drugs that are poorly absorbed after oral administration.Glucomannan is a high
molecular weight water soluble non ionic polymer extracted from the seeds of Araucaria
cunninghamii. Araucaria cunninghamii is a species of Araucaria known as Moreton Bay Pine
or Hoop Pine. Other less commonly used names include Colonial Pine and Richmond River
Pine. The species is found in the coastal rainforests of Eastern Australia and in New Guinea.
The trees can live up to 450 years and grow to a height of about 60 meters. The bark is rough
and splits naturally but does not peel. The leaves on young trees are awl-shaped, 1-2 cm long,
about 2 mm thick at the base and scale-like, incurved, 1-2 cm long and 4 mm broad on mature
trees. The cones are ovoid, 8-10 cm long and 6-8 cm diameter and takes about 18 months to
mature. They disintegrate at maturity to release the nut-like seeds10. Glucomannan is a linear
random copolymer of β (1g 4) that linked D-mannose and D-glucose and approximately one
in 15 of the sugar units11, 12 are acetylated. It is similar to pectin which is not hydrolyzed by
digestive enzyme in human beings and is considered as an indigestible dietary fiber that has
received recognition for reducing the risk of developing diabetes and heart disease. A diet rich
in high-viscosity CGM is generally recommended for improved glucose and lipid profile
control, suggesting a therapeutic potential in the treatment of the insulin-resistance syndrome13.
CGM could be hydrolyzed by ß-mannanase to produce manno-oligosaccharides which plays an
important role in biological systems14. In addition, CGM could form strong, elastic, heat-
stable gels when heated with mild alkali15 and has also been used as drug carrier16, food
preservative17, 18 and for enzyme entrapment19.
*Corresponding author.
D. Saravanan,
Assistant professor,
Ratnam Institute of Pharmacy,
Pidathapolur,
Nellore-525346.
The aim of the present study was to isolate glucomannan from Araucaria cunninghamii and
the characterization of the isolated glucomannan by FTIR and Mass spectroscopy. The isolated
glucomannan was used for preparation of sustained release sodium alginate beads. Sustained
release beads were prepared by using alginate (ALG) and cunninghamii glucomannan (CGM).
The prepared beads were characterized by FTIR and evaluated for swelling index, angle of
repose, tapped density, bulk density, porosity, Carr’s index, SEM analysis and In-Vitro drug
release.
MATERIALS AND METHODS
The seeds of Araucaria cunninghamii were purchased from Bharat Vastu Bhandar (Herbs,
seeds and plants merchants), Dhamawala bazaar, Dehradun, India. Acetic acid, calcium chloride
and sodium hydroxide were purchased from Rankem laboratory, New Delhi, India. Sodium
chlorite was purchased from G. S. chemicals, Bombay, India. Potassium hydroxide was
purchased from M/S Hi-media Ltd., Bombay, India. Boric acid, Barium hydroxide and
Ethanol were procured from Changshu yangyuan chemicals, China. Sodium alginate was
procured from Loba chemi, Bombay, India. Other reagents and solvents used were of analytical/
spectroscopic/HPLC grade as the case desired.
Isolation and purification of glucomannan from Araucaria cunninghamii
The seeds of Araucaria cunninghamii were shade dried and powdered. About 100 gms of dry
powder was suspended with stirring in water (1.67 lts) at 70-80ºC. Acetic acid (11 ml) and
sodium chlorite (33.3 gm) were added for every hour for a total period of 7 hours. The reaction
mixture was cooled and the solid was washed by decantation with tap water. They were then
transferred to a filter and washed with distilled water and ethanol. The white product was then
air dried to yield (78 gm) of holocellulose.
The holocellulose (78 gm) was shaken with aqueous potassium hydroxide (560 ml) at room
temperature for 3 hours. The alkaline extract was removed by filtration through whatmann’s
filter paper and the residue was washed with water. The still wet residue was extracted in the
same way with 17.5% sodium hydroxide containing 4% boric acid. The alkaline extracts and
washings (745 ml) were then poured into ethanol (2.780 l) containing glacial acetic acid (80
ml). The precipitate formed was then washed on the centrifuge with 80% aqueous ethanol,
ethanol and petroleum ether (b.p 30-60ºC).
Crude glucomannan (4 gm) was dissolved in 10% sodium hydroxide (100 ml) and 5% aqueous
barium hydroxide (200 ml) was added with constant stirring over a period of 2 hours. The
precipitate thus formed was collected by centrifuging, washed twice with water, acidified with
acetic acid and poured into ethanol (500 ml). The precipitate was recovered in the usual way.
Obtained product glucomannan was dried at 50ºC for 3 hours and was used for further
characterization. The polysaccharide content (PC) was calculated using the following formula,
PC % = (m1/m2 ) X100
Where m1and m2 are the weight of final white powder and original Araucaria cunninghamii
seeds, respectively20.
FTIR spectroscopy
FTIR spectrum of the raw glucomannan and glucomannan beads were recorded on the FTIR
(Impact 410 spectrometer) under dry air at room temperature using KBr pellets in the range
between 4000 and 400cm”1. The peaks were assigned by comparison with the data reported in
the literature21.

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Mass spectroscopy In-Vitro dissolution studies
Molecular weight of glucomannan was recorded on mass spectrometer, JEOL GC MATE II The drug release study was carried out using USP dissolution test apparatus (XXIII),
paddle type. Study was conducted in 900 ml of dissolution medium maintained at a
Cunninghamii glucomannan preparation temperature of 37 ± 0.5°C with a paddle rotation speed of 100 rpm. The pH of the medium
0.5% (w/v) solution of CGM was prepared in water and the solution was centrifuged to remove was varied over the course of the experiment: 0.1 N hydrochloric acid (pH 1.2) was used
the insoluble material. The supernatant liquid was then poured into an equal volume of for the first 1 hour and of phosphate buffer pH 7.4, till the complete release of drug took
methanol to precipitate CGM. The solid was then filtered, redispersed in water and freeze- place. Samples of 5 ml volume were withdrawn at predetermined time intervals and were
dried. The product got was a fluffy white material, approximately 92% of original powder, replaced with fresh dissolution medium to maintain sink conditions.
which was used for this study.
RESULTS AND DISCUSSION
Formulation of sustained release glucomannan-sodium alginate beads The glucomannan was isolated from Araucaria cunninghamii by a simple method without
The mixture of sodium ALG (1-5% w/v) and CGM (0.5% w/v) were dissolved in deionized using toxic chemicals. The yield was 19%w/w. The isolated glucomannan was characterized
water. Whole solution was then added drop by drop through a needle (0.8 mm hypodermic) by FTIR and Mass spectroscopy.
from a constant height, with a sterilized glass syringe into a beaker containing 100 ml of CaCl2
solution (2% w/v). ALG–CGM beads were hardened in CaCl2 solution for 30 mins and were The FTIR spectrum of the glucomannan in the wavelength range of 4000–400cm”1(Result
then filtered and rinsed with distilled water and dried at 60°C for 3 hours using hot air oven. are shown in Figure-1). In the spectrum of polysaccharide, the wide band observed at
The wet bead diameter was 2.8±0.1 mm as determined microscopically with a vernier scale 3000–3700cm”1could be attributed to the O–H stretching of the glucomannan. The band
micrometer. The composition of glucomannan, sodium alginate and CaCl2 for each batch was at 2922cm “1 was attributed to the asymmetric stretching of C–H, while the band at
shown in Table-1. 1577cm”1 was ascribed to adsorbed water and the bands at 1410 and at 1280cm”1 to the
angular deformation of C–H. The C–O ether bond has shown stretching at about 1162cm”1
Table 1: Formulations while the C–O alcohol bond is shown stretching at 1112 and 1048cm”1. The characteristic
peaks observed at 808–900cm”1were assigned to the ß-pyranose between mannose and
Formulations Sodium alginate Calcium chloride Glucomannan glucose unit. The peak at 1628cm”1 was attributed to carbonyl of acetyl group. These
(%w/v) (%w/v) (% w/w)
results were in agreement with the data reported by Zhang et al. (2001).
G1 1 2 0.5
G2 2 2 0.5 The results indicated that, molecular weight of expected glucomannan (M+) fragments was
G3 3 2 0.5 found to be 666.51 (Result are shown in Figure-2).
G4 4 2 0.5
G5 5 2 0.5

Post formulation studies

Post formulation parameters such as swelling index, angle of repose, tapped density, bulk
density, porosity and Carr’s index of prepared glucomannan beads was determined as per
standard I.P procedure22.

SEM analysis
Blank beads were added to deionized water and lyophilized. For SEM analysis (PHILIPS
XL 30 ESEM, Amsterdam, Netherlands), cross-sections of the freeze-dried blank beads
were obtained using a razor blade. The sections were then coated with gold–palladium for
70s in an argon atmosphere, before observing them under the microscope. The mean bead
diameter and standard deviation were calculated from a sample population of atleast 20
lyophilized beads randomly selected from the population. A partial bead becomes elliptical
in shape due to shrinkage. The average over the short diameter and the long diameter is
considered as the size of the bead.

100.0 Figure. 2: Mass spectrum of glucomannan from Araucaria cunninghamii.

95

90

85

80

75

70

65

60 852

55

50
%T
897
45

40 666

35 2143 617

30

25 1280
1256
20

15 2854
2922 1410 1162
10 1628 1112
3428
1577
5 1048

0.0
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600 450.0
cm-1

Figure.1: FTIR spectrum of glucomannan from Araucaria cunninghamii

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The post formulation studies of glucomannan sustained release beads were carried out as The SEM micrograph of the beads of ALG-CGM (Result are shown in Figure-5, 6, 7 and
per I.P. The results indicate that the bulk density, tapped density, carrs index, angle of 8). The surface of ALG-CGM bead was smooth and had clear cylindroids dents (Figure-
repose and porosity was found to be within the limits. The values obtained for the post 5). The above phenomenon indicated that the phase separation happened when mixture of
formulation parameters for the formulations G 1to G5 (Result are tabulated in Table-2 and sodium ALG and CGM was dropped into a cross linking solution containing Ca2+. After
Table-3). gelling for a while, beads were rinsed with distilled water. At this time, CGM was lost
from the surface of beads were created. At the inner of the beads, homogenous and
Table 2: Post formulations study of glucomannan-sodium alginate beads
honeycomb like open cavities were formed in ALG beads (Fig. 6), while a compact core
Batch Bulk density Tapped density Porosity Carr’s index Angle of repose was found in ALG-CGM bead and separated partly from the surface due to the inner phase
(g/cc) (g/cc) (%)
(?) separation. The surface morphology of beads shown in Figure-7 and 8.
G1 0.5210 0.6314 12.50 12.56 15.500
G2 0.5322 0.6132 13.10 12.10 16.250
G3 0.5434 0.6250 13.04 13.12 16.481
G4 0.5208 0.5952 12.50 12.60 14.530
G5 0.5421 0.6015 12.55 13.05 16.120

Table 3: Comparisons of swelling index properties and yield of different formulations

Formulations Swelling index (%) Yield (%)

G1 98 90
G2 97 92
G3 98 85
G4 99 98 Figure. 5 Figure. 6
G5 99 96

We use IR spectra to discuss whether CGM exits within beads after gelling and the
interaction between carbohydrates. Figure-3 and Figure-4 give the IR spectra of the beads
at the range of 4000–900 cm”1. In ALG–KGM beads, some peaks disappeared or became
weak due to interaction or superposition between groups of ALG and CGM. The stretching
of —CH at 2922.5 cm”1 in ALG and that of at 2922 cm”1 and 2854 cm”1 in CGM
disappeared. The carbonyl of aceto groups at 1628 cm”1 of CGM also cannot be seen. The
absorption band at 1627 cm”1 of COO— in ALG may be coupled with the absorption band
at 1577 cm”1 of intra-molecular hydrogen bonds in CGM and forms a new large peak at
1738 cm”1. A weak peak at 1435 cm”1 was formed due to merging of 1460 cm”1 of ALG and Figure. 7 Figure. 8
1410 cm”1 of CGM. A peak at 1048 cm”1 in CGM is disappeared. Moreover, the absorption
band at 1384 cm”1 in ALG interacts with CGM and forms a broad peak at about 1365 cm”1. Figure. 5, 6, 7 & 8: Scanning electron micrograph of glucomannan-sodium alginate beads of
The above phenomenon indicated that CGM was contained within beads and faintness Araucaria cunninghamii
hydrogen binding and electrostatics interaction exists between ALG and CGM. To study the effect of sodium alginate concentration on glucomannan release, the sodium
alginate was used at 5 different concentrations: 1(G1), 2 (G2), 3 (G3), 4 (G4), 5 (G5) %w/
v. The release profiles of these formulations (Result are shown in Figure-9 and Table-4).
The results indicated the more sustained effect with all increase in concentrations of
sodium alginate. The alginate disintegration was monitored by the exchange of Ca2+ and
Na+ in the dissolution medium. The sodium alginate concentration in the formulation
greatly influenced the steady state release of glucomannan from alginate beads. Increased
alginate gel density per unit volume was also thought to affect the decreased pore size
within the gels and thus glucomannan release becomes slow.

Figure. 3: FTIR spectrum of glucomannan-sodium alginate beads

Figure. 9: Percentage drug release profile of glucomannan-sodium alginate beads

Table-4: Comparative % drug release profiles of all formulations of glucomannan-sodium

alginate beads

Time(hrs) Buffer G1 G2 G3 G4 G5

1 0.1 N HCl 0 0 0 0 0
2 7.4 pH phosphate 34% 30% 24% 19% 18%
3 Buffer 58% 45% 41% 33% 25%
4 72% 63% 60% 55% 40%
5 98% 85% 74% 61% 58%
6 - 99% 89% 75% 72%
7 - - 99% 90% 86%
8 - - - 100% 98%

Figure. 4: FTIR spectrum of sodium alginate beads

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CONCLUSION
A simple method without using toxic chemicals for isolating cunninghamii glucomannan
from Araucaria seeds has been investigated. The cunninghamii glucomannan from Araucaria
seeds was white, hard to dissolve in water and its content was about 19% (w/w). The
molecular weight of glucomannan from Araucaria cunninghamii was 666.51 and prepared
beads showed the higher degree of swelling and slow drug release was found for beads of
formulation G5 (5% w/v sodium alginate and 2% w/v calcium chloride). Thus, the
concentration of polymer had major influence on swelling process, matrix integrity. Hence
from the above results it can be proved that linear relationship exists between swelling
process and concentration ratio of sodium alginate. From the in- vitro drug profiles, it was
found that drug release rate decreased as the concentration of polymers sodium alginate
increases. The cunninghamii glucomannan from Araucaria species in eastern Australia and
in New Guinea exhibited a potential application as both a food and medicine for human.
ACKNOWLEDGEMENTS
We are grateful thanks to our principal Prof. M. Gobinath, Ratnam institute of pharmacy,
Pidathapolur, Andhra Pradesh- 524 346 for providing all facilities to completion of this
research work.
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