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1. Introduction
Nanotechnology is a promising approach of twenty-first century. It is an attractive area of
research related to production of nanoparticles of variable size and shape, in addition to
their potential advantages in clinical medicines.[1] It is on doorstep of providing a host of
new objects and approaches in modernising therapeutic and pharmaceutical field. In
present era, it significantly promotes the formation of biological medicine and
bioavailability enhancement of phytomedicines. Nanonisation of herbal drugs is opening
the new epoch of herbal drug discovery.[2]
Herbal medicines are extensively used all over the world since ages. The efficacy of
several species of therapeutic plants relies on the release of biologically active compounds.
The majority of the active components of extracts are incapable to pass the lipid
*Corresponding author. Email: nazishjahanuaf@yahoo.com
2015 Taylor & Francis
N. Jahan et al.
membranes of the cells because either they have markedly high molecular size or poor
water solubility, thereby suffers from low absorption and poor bioavailability.[3] Many
phytomedicines, thereby due to their poor absorption, exhibit least or no considerable
in vivo activity despite of their amazing in vitro potential. Due to these obstacles, some
extracts are not used clinically. It has been extensively suggested to incorporate herbal
drugs with nanotechnology, as nanostructured systems might be capable to strengthen the
action of herbal extracts decreasing the necessary dosage and side effects, and improving
bioactivity.[4] Numerous nano-oriented approaches are being projected with the aim of
optimising the technological features of drugs.[5,6] Nanosuspension technology is
introduced as a result of countless efforts made by formulation scientists and has been
evolved as a potent candidate for the delivery of the poorly soluble drugs in a more
efficient and pronounced manner.[7] Because by considering the limitations of previously
used technologies of particle size reduction, pharmaceutical scientists have been waiting
for such a universal approach that would be able to address the formulation-related
problems and barriers of poorly soluble drugs up to their desired level.[8,9]
Silybum marianum commonly known as milk thistle is a member of Asteraceae family
and genus Silybum. Silymarin is the major bioactive compound isolated from its seed
which has innumerable applications. Silymarin is employed for the oral therapy of chronic
liver disorder but it has poor aqueous solubility thereby poor bioavailability.
Pharmacokinetic studies have revealed that after oral administration just 23% 47% of
silymarin enters the systemic circulation.[10] Elettaria cardamomum generally known as
cardamom has well-known culinary value. Seeds of this plant are used in folk medicines
for the treatment of cardiac and gastrointestinal disorders.[11] Coriandrum sativum
commonly known as coriander has various pharmacological properties. Coriander is used
as anti-inflammatory, antihypertensive, antiedemic, antiseptic, antidiabetic, lipolytic and
myorelaxant.[12] This project had been designed to prepare the nanosuspension of three
poor aqueous soluble plants extract to enhance their bioactivities. Nanosuspensions of
S. marianum, E. cardamomum and C. sativum were prepared by nanoprecipitation method.
Prepared nanosuspensions were characterised for their particle size and morphology by
using scanning electron microscopy (SEM).
2. Experimental section
2.1. Chemicals and reagents
Polyvinyl alcohol (PVA), 2,2-diphenyl-1-picrylhdrazyl (DPPH), ascorbic acid, butylated
hydroxyl toluene (BHT), nitroblue tetrazolium regent, hydroxylamine hydrochloride,
ammonium thiocyanate and linoleic acid were purchased commercially from SigmaAldrich (St. Louis, MO, USA). All other chemicals were of analytical grade of standard
Merck (Darmstadt, Germany).
N. Jahan et al.
Figure 1. SEM image of particle size (a) and morphology (b) of Silybum marianum nanosuspension.
Figure 2. SEM image of particle size (a), (b), (c), (d) and morphology (e) of Elettaria cardamomum
nanosuspension.
N. Jahan et al.
Figure 3. SEM image of particle size (a) and morphology (b) of Coriandrum sativum
nanosuspension.
terms of IC50 value (Table 1) that indicates the concentration of antioxidant required to
neutralise 50% free radicals. Lower value of IC50 thereby indicates high inhibitory effect.
DPPH scavenging activity is a well-known method to determine the antioxidant potential
of medicinal plants in short time period.[18] Among plants extract, lowest IC50 value for
DPPH free radical scavenging assay was shown by E. cardamomum (5.21 0.1 mg/ml).
Nanosuspension of C. sativum (0.59 0.01 mg/ml) exhibited minimum IC50 value. Nitric
oxide radicals overproduction is also said to be related to different types of
neurodegenerative and muscles diseases. Nitric oxide radical scavenging potential of
different plants and their nanosuspension was determined by the generation of nitric
oxides radical in vitro by sodium nitroprusside. These free radical lead to the production of
nitrite ions by reacting with oxygen. Nitric oxide scavengers compete with oxygen and
thereby diminish the formation of nitrites ions.[19] S. marianum nanosuspension showed
Table 1. IC50 value of different plants formulations and standards.
IC50 value (mg/ml)
Plants formulation
and standards
DPPH radical
scavenging
Nitric oxide
radical scavenging
Superoxide
radical scavenging
13.62 1.02
1.95 0.09
5.21 0.1
1.74 0.07
16.51 1.22
0.59 0.01
2.43 0.2
3.49 0.8
7.73 0.11
0.34 0.02
7.99 0.1
2.14 0.4
5.63 0.23
1.32 0.03
1.22 0.01
2.69 0.2
8.15 0.4
1.19 0.31
4.0 0.1
1.63 0.08
4.81 0.22
0.81 0.11
2.6 0.07
2.02 0.11
lowest IC50 value (0.34 0.02 mg/ml) for this assay. Superoxide radical is regarded as a
powerful biological cause of reactive oxygen species because it leads to formation of
potent and hazardous hydroxyl radicals.[20] For this assay, lowest IC50 value was also
shown by C. sativum nanosuspension (0.81 0.11 mg/ml). Result depicted in Table 1
clearly reflected that nanosuspensions of plants showed more scavenging of free radicals
than crude plant extracts. Nanosuspensions of plants even show better results than
standard compounds BHT and ascorbic acid because when particle size is reduced up to
nano range, not only surface area but concentration gradient is also increased which
results in dramatic increase of the dissolution velocity as compared to a micronised
product and activity of extracts was improved by formulating their nanoparticles.[21]
Moreover, the use of surfactant and continuous stirring helped the nanosuspension
particles to avoid agglomeration, thus avoid the Ostwalds ripening [22] and led to
enhanced activity of drug. In the earlier work, same relation of particle size reduction and
drug activity was demonstrated by some scientists.[23,24] Thus, it can be stated that
particle size reduction of plants extract significantly increased their in vitro antiradical
potential which might lead towards their better in vivo activity. Lipid peroxidation that is
generally known as primary toxicological event is caused by the formation of free radicals
from a diversity of sources. It leads to oxidative degradation of lipids thereby damaging
cellular membranes. Ammonium thiocyanate method was used to evaluate the anti-lipid
peroxidative effects of plants extract [25] and nanosuspensions and results were reported
in terms of percentage inhibition (Figure 4). Nanosuspensions of all plants exhibited more
percentage inhibition of lipid peroxidation than their crude extracts. Percentage inhibition
for S. marianum nanosuspension was found to be 83.76 1.1% as compared to its crude
extract (39.76 5.01%). Nanosuspenion of C. sativum was found to be more effective
towards inhibition of lipid peroxidation after 72 h (91.75 2%) among all plants extract,
N. Jahan et al.
nanosuspensions and standards. BHT and ascorbic acid exhibited considerable amount of
percentage inhibition but not more than plants nanosuspension. Appreciable inhibition of
peroxidation might be ascribed to the presence of well-known antioxidants, for instance
flavonols, xanthones, di-anthraquinones and flavans potentially responsible for the
considerable activity of the extracts. Due to nanosizing, surface area was increased and
molecules interacted very rapidly with solvent molecules which resulted in increased
solubility. Therefore, results of inhibition of lipid peroxidation might be based on
enhanced solubility of the samples. The crude extract of plants and standards due to larger
particle size were failed to scavenge free radicals in manner similar to that of
nanosuspension.
4. Conclusion
It is concluded that nanosuspensions of the selected plants S. marianum, C. sativum and
E. cardamomum significantly enhanced the antiradical potential as compared with their
crude extracts.
Disclosure statement
No potential conflict of interest was reported by the authors.
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