Industrial Crops and Products 44 (2013) 290293

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Industrial Crops and Products

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Alginic acid and derivatives, new polymers from the endangered Pancratium
maritimum L.
Adnen Sanaa a, , Abdennacer Boulila b , Mohamed Boussaid a , Najeh Ben Fadhel a
a
National Institute of Applied Sciences and Technology (INSAT), Department of Biology, Laboratory of Plant Biotechnology, North Urban Center, BP 676, 1080 Tunis Cedex, Tunisia
b
National Institute of Physico-chemical Research and Analysis, Laboratory of Natural Substances, Sidi Thabet 2020, Ariana, Tunisia

a r t i c l e i n f o a b s t r a c t

Article history: Alginic acid and derivatives were reported for the rst time from the bulb of Pancratium maritimum
Received 3 October 2012 L. The brown algae Sargassum muticum was used as a source of sodium alginate. The alginic acid and
Received in revised form 9 November 2012 calcium alginate production methods were performed. FT-IR spectra of sodium alginate extracted from
Accepted 11 November 2012
P. maritimum reported an intense peak around 1614 cm1 and a second intense peak around 1431 cm1 .
Our analysis showed that the sodium alginate characteristic peaks of P. maritimum and S. muticum were
Keywords:
similar. Chemical data in our study can provide some insight into the bioactive molecules in the species
Alginic acid
with important industrial and pharmaceutical uses and may lead to use P. maritimum as a new source of
FT-IR
Pancratium maritimum L.
sodium alginate.
Sodium alginate 2012 Elsevier B.V. All rights reserved.

1. Introduction et al., 1998), pancratistatine (Pettit et al., 1995), N-dimethyl-

Pancratium maritimum L. (Amaryllidaceae) is distributed in the
Mediterranean, the Atlantic, the Black and Caspian coasts (Dothan,
1986). The habitat of P. maritimum in the immediate vicinity of
the sea is characterized by direct exposure to sea breezes and to
continuous sprays by high air humidity and by strong radiation
(Eisikowitch and Galil, 1971). The species is severely threatened
in its original range, the sandy coasts of the Mediterranean Sea by
over collection, urbanization and tourism development (Sanaa and
Ben Fadhel, 2010). The plant has attracted attention as a source of
valuable acaricidal, insecticidal (Abassy et al., 1998) and antifungal
activities (Sur-Altiner et al., 1999).
Many alkaloids have been reported for this species such as dihy-
drolycorine, norpluviine (Sandberg and Michel, 1968), lycorine,
6-O-methylhemanthidine, O,N methylnorbelladine, hippeastrine,
hordenine, harbanthine, ungiminorine, ungiminorine-N-oxyde,
vitattine (Tato et al., 1988), tazettine, pancracine, lycorinine, galan-
thamine, sickernbergine, homolycorine, trispheridine, heman-
thamine, pseudolycorine, 9-O-dimethylhomolycorine, 11-hydrox-
yvitattine, ungiminorine, zefbetaine, narciclasine-4-O--d-glucop-
yranoside (Abou-Donia et al., 1991), 3,11-dihydroxy-1,2-dehyd-
rocrinane, buphanisine, crinine,3-methoxy-6-dihydroxy-3-me-
thoxy-1,2-dehydrocrinane, 8-hydroxy-9-methoxycrinine (Sener
Corresponding author. Tel.: +216 71703829; fax: +216 71704329.
E-mail address: sanaaadnen@yahoo.fr (A. Sanaa).
galanthamine, lycoramine (Youssef and Frham, 1998), pancrita-
mine, acetyllycoramine, N-dimethyllycoramine (Youssef, 1999),
graciline, 6 dioxytazettine, galanthane, dimethylmarithidine and
crinane-3-one (Berkov et al., 2003).
Some alkaloids have shown anti-tumor (pancratistatine and
ungiminorine: Pettit et al., 1995), anti-viral (lycorine), anti-
cholinesterase (galanthamine) and analgesic activities (galan-
thamine and lycorine: Torras-Claveria et al., 2013). Besides, the
major components of ower volatiles of the species were hep-
tacosane, hexadecanoic acid, benzyl benzoate, octacosane and
hexacosane (Sanaa et al., 2012).
Recently, a great deal of interest has been developed to isolate
novel bioactive compounds from coastal plants with industrial and
pharmaceutical interests such as alginic acid and derivatives. In
fact, they are largely used in food, in textile and in pharmaceutical
industries (Littlecott, 1982). Our aim in this work is to isolate and
identify alginic acid and derivatives from P. maritimum.
2. Materials and methods
Samples of P. maritimum were collected from Bizerte coast in the
north of Tunisia (37 18 37.68 N, 9 51 24.24 E). A voucher speci-
men (P.MAR 12) was deposited at the herbarium of the National
Institute of Applied Sciences and Technology. In this study, the
brown algae Sargassum muticum was used as source of sodium
alginate.

0926-6690/$ see front matter 2012 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.indcrop.2012.11.014
 A. Sanaa et al. / Industrial Crops and Products 44 (2013) 290293 291

Fig. 1. Flow chart for the production of sodium alginate.

After McHugh et al. (2001).

3. Experimental
P. maritium and S. muticum were submitted to the same steps of
sodium alginate extraction. Bulb (50 g) of P. maritimum and 50 g of
S. muticum were broken each into pieces and stirred with a solution
of 100 ml Na2 CO3 (1.5%) at 65 C. HCl (5%) was added to the ltered
extract to pH 12. Alginic acid formed gelatinous pieces that must
be separated from the water. Ethanol was added to give a 50:50
mixture of alcohol and water to precipitate the alginic acid. Then
a solution of NaOH (40%) was added gradually until the resulting
paste reaches pH 67 (Venegas et al., 1993).
Alginates were present in the cell walls of brown algae as the cal-
cium, magnesium and sodium salts of alginic acid (Lee et al., 2011).
The goal of the extraction process was to obtain dry, powdered,
sodium alginate. The rationale behind the extraction of alginate
from the seaweed and P. maritimum was to convert all the alginate
salts to the sodium salt, dissolve this in water, and remove the sea-
weed and P. maritimum residues by ltration. The alginate must
292 A. Sanaa et al. / Industrial Crops and Products 44 (2013) 290293

then be recovered from the aqueous solution. There were two dif-
ferent ways of recovering the alginate. The rst was to add acid to
form alginic acid. This did not dissolve in water and the solid alginic
acid was separated from the water. The alginic acid was separated
as a soft gel and some of the water must be removed from this. After
this has been done, alcohol was added to the alginic acid, followed
by sodium carbonate which converts the alginic acid into sodium
alginate (Na Alg). This did not dissolve in the mixture of alcohol and
water, so it could be separated from the mixture, dried and milled
to an appropriate particle size that depends on its particular appli-
cation. The second way of recovering the sodium alginate from the
initial extraction solution was to add a calcium salt. This caused
calcium alginate to form with a brous texture; it did not dissolve
in water and can be separated from it. The separated calcium algi-
nate was suspended in water and acid was added to convert it into
alginic acid. This brous alginic acid is easily separated, placed in a
planetary type mixer with alcohol, and sodium carbonate is grad-
ually added to the paste until all the alginic acid is converted to
sodium alginate (Na Alg).
The sodium alginate (Na Alg) extracted from the brown algae Fig. 2. FT-IR spectra of sodium alginate from P. maritimum and S. muticum.
was used to the comparative FT-IR analysis with the sodium algi- , FT-IR spectra of sodium alginate of Sargassum muticum; , FT-IR spectra of sodium
nate extracted from the bulb of P. maritimum. The paste of sodium alginate of Pancratium maritimum.
alginate (Na Alg) can be extruded as pellets, oven dried and milled
for further spectral analysis with a Perkin-Elmer FT-IR Spectrum
1000 apparatus in the 4000400 cm1 region. These essentials of
the process were illustrated in the ow diagram in Fig. 1. 4.3. FT-IR analysis

FT-IR spectroscopy has been used to characterize alginic acid

4. Results and discussion
4.1. Alginic acid method
First, alginic acid was formed in soft, gelatinous pieces that must
be separated from the water. Filtration was not possible because of
the soft jelly-like nature of the solid. Second, an excess of sodium
carbonate was used in the original extraction, this was still present
in the ltered extract so that when acid was added, carbon diox-
ide will be formed. Fine bubbles of this gas were attached to the
pieces of alginic acid and lifted them to the surface where they
were continuously scrapped away. Moreover, the mass of alginic
acid contained 9899% water. This water content must be reduced.
Centrifuging increased the solids to 78% and this was sufcient.
The 78% alginic acid was placed in a mixer and, allowing for the
water contained in the alginic acid, enough alcohol was added to
give a 50:50 mixture of alcohol and water. Then solid sodium car-
bonate was added gradually until the paste of sodium alginate was
extruded as pellets, oven dried and milled. Our results were com-
parable to those observed by McHugh et al. (2001).
4.2. Calcium alginate method
and derivatives (Mackie, 1971). This analysis showed that the
sodium alginate characteristic peaks of P. maritimum and S.
muticum were similar (Fig. 2). FT-IR spectra reported an intense
peak around 1614 cm1 and a second intense peak around
1431 cm1 . Our results were in agreement with the studies
done by Huang et al. (1999), who described two intense peaks
around 1614 cm1 and 1431 cm1 respectively. Moreover, in the
IR alginic acid presented bands at 1730 and 1631 cm1 , while
sodium alginate presented a doublet at 1614 and 1431 cm1 ,
evidencing the presence of salied carboxyl groups (Chandia et al.,
2001).
In conclusion, P. maritimum L. is mainly grown on sandy
Mediterranean coasts. Populations were at present endangered
and represented by scattered individuals as a result of habitats
destruction. This study is the rst assay to identify alginic acid and
derivatives in the species. They are largely used in cosmetics, in
pharmaceutical and in food industries. From a chemical point of
view, the alginate molecule may look very simple. This simplica-
tion of its chemical structure may lead potential commercial users
to treat it as a commodity. Adding to that, chemical data in our study
can provide some insights to use P. maritimum as a new source of
sodium alginate and the cultivation of the species could be of great
interest.

Our ndings were similar to those obtained by Romero- Acknowledgements

Gonzalez et al. (2001). Calcium alginate was formed after adding
calcium chloride to the ltered extract. The calcium alginate was
formed as bers. Besides, this brous material was separated on a
metal screen and washed with water to remove excess calcium.
It was then stirred in dilute acid and converted to alginic acid,
which retained the brous characteristics of the calcium alginate.
Then solid sodium carbonate was added gradually until the paste
of sodium alginate was extruded as pellets, oven dried and milled.
The disadvantage of this second method, compared to the alginic
acid method, was that an extra step is added to the process. The
advantage was that the handling of the brous calcium alginate and
alginic acid was much simpler and alcohol was not needed (Horn
et al., 2000).
This research was supported by a grant of the Ministry of Scien-
tic Research and Technology (Research grant 99/UR/09-10). We
wish to express our thanks to Dr. Fatma Karoui for proof reading
the manuscript.
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