Research Article
ejbps, 2015, Volume 2, Issue 6, 162-165.
SJIF Impact Factor 2.062
ISSN 2349-8870
Volume: 2
European Journal of Biomedical and Pharmaceutical Sciences
Issue: 6
Pharmaceutical sciences
162-165
Year: 2015
http://www.ejbps.com
European Journal of Biomedical
Shashank et al.
AND
EVALUATION THE QUANTITY OF TANNINS IN AILANTHUS EXCELSA ROXB.
Bhatt Shashank1*, Dhyani Suresh2, Kumar Sandeep3
1,3
Department of Biotechnology, NIET, NIMS University, Jaipur, Rajasthan- 303121 India
2
Himgiri Zee University, Dehradun, Uttarakhand- 248197, India.
*Author for Correspondence: Bhatt Shashank
Department of Biotechnology, NIET, NIMS University, Jaipur, Rajasthan- 303121 India.
,
Article Received on 20/08/2015
Article Revised on 12/09/2015
Article Accepted on 05/10/2015
ABSTRACT
The leaves of Ailanthus excelsa Roxb. is a good source of tannins. The quantity of tannins differ in root, bark, and
leaves. The tannins were extracted from leaves collected from different places in Mandsaur District Mandsaur
(Madhya Pradesh). The percentage of tannins’ content was calculated with standard tannic acid and found 18.4,
18.0, 19.2, 18.8 and 17.6% respectively. The S.Em. value 0.163299 and CD 5% 0.481733 were found.
KEYWORDS: Ailanthus excelsa Roxb., Secondary Metabolites, Tannins.
INTRODUCTION
Plants play an important role in most of the organisms
that are dependent upon directly or indirectly. These
have an important place in the medicinal world since
ancient times. The traditional herbal plants have been a
great source of therapeutic agents. In Aurveda, not only
within India but also outside, has revived ancient
traditional methods have been revived. Medicinal plants
have been treated as an important source of healthcare.
They have been an important part of international trade
and commerce. The medicinal plants have been
recognized as non-narcotic due to less side effects and
easy availability at low costs.
The world Health Organization (WHO) has estimated
that near about 80% population of developing countries
depend on traditional medicines for primary healthcare.
These medicines are prepared by plants’ bark, roots,
fruits and leaves which play a major role in the
preparation of medicines. Herbal medicines are prepared
from plant leaves, stems, roots, bark etc. These materials
contain active ingredients that are used for treatment of
chronic ailments. Plants can be considered into medicinal
category if they possess pharmacological activities.
Approximately, 15,000-20,000 plant species have good
medicinal values. Gum, resins and latex, which are
excretory parts of plants are used in the preparation of
drugs (Khan et al., 2011). The primary metabolites have
essential role in photosynthesis, respiratory, growth and
development. Secondary metabolites are not directly
involved into normal growth, development (Schanderi,
1970). These secondary metabolites play a major role in
defense mechanism against herbivores (Ehrlich et al.,
1964; Fraenkel, 1959; Whittaker et al., 1975). Various
www.ejbps.com
effective contents are present in the medicinal plants.
Alkaloids, flavonoids, glycosides, tannins, saponins etc.
are the most effective contents against various diseases.
These contents are also used for the development of new
medicines by which people can develop their immune
system.
Tannins are polyphenolic secondary metabolites of
higher plants. Tannin containing plants’ extracts are also
used in the treatment of various diseases such as diarrhea
(Yoshida et al., 1991), diuretics (Okuda et al.,1983;
Hatano, et al., 1991), stomach and duodenal tumors
(Saijo et al., 1989), anti-inflammatory, antiseptic, and
haemostatic pharmaceuticals (Haslam, 1989). They are
also used in the production of ink (iron gallate ink) and
used to clarify wine, beer, and fruit juices (Wurdig et al.,
1989). Ailanthus excelsa Roxb. (Fig. 2) is one of the
important plants that show medicinal properties. The
Plant belongs to Simaroubaceae family and its common
name is Mahanimba due to its resemblance with neem
tree (Azadirachita indica). It is also called Maharukha
due to its large size. The generic name ‘Ailanthus’ comes
from ‘ailanthos’ that means ‘Tree of Heaven.’
Ailanthus excelsa Roxb. is one of the medicinal plants
that is used in the Indian System of medicines. Its parts
are used, as Folk medicine, in the treatment of several
ailments in which asthma, cough, colic pain, cancer,
diabetes, diarrhea, dysentery, cholera and microbial
infections are included (Kritikar et al., 1995).
162
Shashank et al.
European Journal of Biomedical and Pharmaceutical Sciences
MATERIAL AND METHODS
Collection of Plant Material
The fresh leaves (Fig.3) of Ailanthus excelsa Roxb.
belonging to the family Simaroubaceae, were collected
from five different places in Mandsaur, districts
Mandsaur, Madhya Pradesh (Fig.1). These places were
indicated by M-1, M-2, M-3, M-4 and M-5. The District
Mandsaur is situated at the northern projection of
Madhya Pradesh between the parallels of latitude 23 0 45'
50" North and 250 2' 55" North, between the meridians of
longitude 740 42' 30" East and 750 50'20" East. The plant
material was taxonomically identified, confirmed and
authenticated by Dr. Rakesh Mohan Painuli, Incharge,
Herbarium, Department of Botany, HNB Garhwal
University, Srinagar (Garhwal), Uttarakhand The
identification code is (HNB 20705). The collected green
leaves were shade dried and then the dried material was
crushed to coarse powder in grinder. The powder was
stored in an airtight container for extraction.
Figure 1- Map of Madhya Pradesh.
Figure 2- Photograph of Ailanthus excelsa Roxb.
Plant.
Figure 3- Photograph of Ailanthus excelsa Roxb. leaf
collected from Mandsaur District Mandsaur
(Madhya Pradesh).
Preliminary Screening of Secondary Metabolites
In the previous research publication, it had been proved
that the tannin contents are present in the leaf part of
Ailanthus excelsa Roxb. The gelatin and lead acetate
tests were performed for the identification of tannins in
95% ethanolic extract (Bhatt s, et al. 2012).
Quantitative Estimation of Tannins
Estimation of tannins was carried out by using FolinDenis reagent [Anonymous, 1980]. Tannin-like
compounds were reduced into phosphotungstomolybdic
acid in alkaline solution to produce a highly colored blue
solution, the intensity of which was proportional to the
amount of tannins. Weighed 500 mg sample of each
place was transferred into 250ml conical flasks. After
adding 75 ml water, it was heated gently in a flask and
boiled for 30min. The samples were centrifuged at 2000
rpm for 20 minutes and collected the supernatants in
each 100ml volumetric flask and made up the volume.
Transferred 1ml of the each sample from 100ml
volumetric flask and added 75ml water. 5ml Folin-Denis
reagent was added and mixed 10ml sodium carbonate
solution and diluted it with distilled water and volume
made upto 100 ml., shaked well and read the absorbance
at 700 nm after 30 minutes. The datas were calculated by
standard tannic acid curve.
RESULT AND DISCUSSION
Generally, most of the parts of a plant such as in the
bark, wood, leaves, fruit, roots and seeds have high
concentration of tannins. Tannins are the effective
compounds that have good antibacterial activity against
various bacterial species. The quantities of tannins were
determined with Folin-Denis method and percent
quantities were calculated by standard tannic acid graph
(Fig. 4A). The standard tannic acid and sample readings
were mentioned respectively in table-1and 2.
The five replicates of Ailanthus excelsa Roxb. were
taken and calculated the quantities of tannins content in
percentage. The percent tannin contents- 18.4%, 18.0%,
19.2%, 18.8% and 17.6% were found in five replicates.
www.ejbps.com
163
Shashank et al.
European Journal of Biomedical and Pharmaceutical Sciences
M-3 indicated higher percent quantity of tannins in the
plant leaf compared to others then M-4, M-1, M-2 and
M-5 showed decreasing order quantities. These datas
were calculated in two ways ANOVA-CRD and found
their significant difference among the replicates. These
differences are due to microclimatic conditions and soil
category in which the plants grow. The graphical
presentation of the various tannin contents (%) are given
in figure 4 (B).
antimicrobial agent and water-soluble plant polyphenols
that are precipitate proteins. These contents precipitate
proteins of microorganism by which they control the
growth of microorganisms. Tannins have various
physiological
activities
such
as
anti-irritant,
antisecretolytic, antiphlogistic, antimicrobial and
antiparasitic effects.
Tannins are generally found in different parts of plant
and used in leather industries. These are good
Table- 1: Quantitative Estimation of Standard Tannic Acid
No. of
Tannic Acid Solution
Distilled
Folin- Denis
Tubes
(1 mg/ml) ml.
Water(ml)
Reagent (ml)
Blank
0.000
0.50
0.25
T1
0.010
0.49
0.25
T2
0.020
0.48
0.25
T3
0.030
0.47
0.25
T4
0.040
0.46
0.25
T5
0.050
0.45
0.25
T6
0.060
0.44
0.25
T7
0.070
0.43
0.25
T8
0.080
0.42
0.25
T9
0.090
0.41
0.25
T10
0.100
0.40
0.25
Sodium
Carbonate (ml)
1.25
1.25
1.25
1.25
1.25
1.25
1.25
1.25
1.25
1.25
1.25
O.D. at
700 nm
0.000
0.068
0.175
0.393
0.278
0.360
0.429
0.573
0.571
0.673
0.775
Tannic
Acid Conc.
0.0
0.02
0.04
0.06
0.08
0.10
0.12
0.14
0.16
0.18
0.20
Table –2: Quantity of Tannin Contents (%) in Leaves of Ailanthus excelsa Roxb. of Mandsaur (M.P.)
Replication
Extract of Ailanthus
Folin- Denis
Sodium
O.D. at
Content
No.
excelsa Leaves (ml.)
Reagent (ml)
Carbonate(ml)
700 nm
(%)
M-1
1 ml
0.25
0.75
0.046
18.4
M-2
1 ml
0.25
0.75
0.045
18.0
M-3
1 ml
0.25
0.75
0.048
19.2
M-4
1 ml
0.25
0.75
0.047
18.8
M-5
1 ml
0.25
0.75
0.044
17.6
S.Em.
0.163299
CD 5%
0.481733
CD 1%
0.657103
SIG/NS
SIG
Figure 4 (A) - Standard Tannic Acid Curve
www.ejbps.com
Figure 4 (B) - Curve of Different Quantities of Tannin
Contents in Leaves of Ailanthus excelsa Roxb.
Collected from Mandsaur (M.P.)
164
Shashank et al.
European Journal of Biomedical and Pharmaceutical Sciences
CONCLUSION
These datas indicated the quantities of tannins in leaves
of Ailanthus excelsa Roxb. that were collected from
different places. The environmental factors as soil
quality, water source, atmosphere etc. are effective in the
quantities of primary and secondary metabolites.
Therefore, the quantities differ from each other but our
all study indicated that the quantities of tannins in shade
dried leaves were good and it will increase in green
leaves of Ailanthus excelsa Roxb. Thus, it was concluded
that this plant can used in the treatment of various
diseases.
9.
10.
11.
12.
13.
ACKNOWLEDGEMENT
Praying and dedicating my research article to Maa
Saraswati, the goddess of knowledge
and wisdom, I
am unable to find words to express my deepest gratitude
to my parents, Mr. Krishna Kumar Bhatt and Mrs.
Subhadra Bhatt whose encouragement helped me to go
ahead on this bright path. I share the credit of my work to
respectable elder brother, Mr. Mayank Bhatt who very
often guided me in the work.
14.
Okuda T, Hatano T, Yazaki K. Praecoxin B, C, D
and E. Novel ellagitannins from Stachyurus praecox.
Chem. Pharma. Bull., 1983; 31(1): 333-336.
Saijo R, Nonaka GI, Nishioka I. Tannins and related
compounds. LXXXIV. Isolated and characterization
of five new hydrolysable tannins from the bark of
Mallotus japonicas. Chem. Pharma. Bull., 1989;
37(8): 2063-2070.
Schanderi SH. Methods in Food Analysis: Academic
Press, New York., 1970; 709.
Whittaker RH. Communities and ecosystems, 2d ed.,
Macmillan Publishing Co. New York, 1975; 385.
Würdig G, Woller R. Chemie des Weines. Stuttgart,
Eugen Ulmer GmbH and Co., 1989; 926s.
Yoshida T, Ohbayashi H, Ishihara K, Ohwashi W,
Haba K, Okano Y, Shingu T, Okuda T. Tannins and
related polyphenols of melastomataceous plants. I.
Hydrolyzable
tannins
from
Tibouchina
semidecandra COGN. Chem. Pharma. Bull., 1991;
39: 2233-2240.
The research work would have only been a dream, had
my way not been enlightened, by my well wishers and
the above respectables. Last but not least, the Almightly
God is unforgettable without whose kindness and grace
nothing could have happened.
REFERENCE
1. Anonymous. Official methods of analysis. 13th
edition. Association of official agricultural chemists,
Washington, D.C., 2004; 1980.
2. Bhatt S, Dhyani S. Preliminary phytochmical
screening of Ailanthus excelsa Roxb. Int J Curr
Pharm Res., 2012; 4(1): 87-89.
3. Ehrlich PR, Raven PH. Butterflies and plants: a
study of coevolution. Evolution (Society for the
Study of Evolution)., 1964; 18(4): 586-608.
4. Fraenkel GS. The raison d’être of secondary plant
substances: these odd chemicals arose as a means of
protecting plants from insects and now guide insects
to food. Science., 1959; 129(3361): 1466-70.
5. Haslam E. Plant Polyphenols-Vegetable Tannins
Revisited- Chemistry and Pharmacology of Natural
Products, First Published, Cambridge University
Press, Cambridge., 1989; 165.
6. Hatano T, Yazaki K, Okonogi A, Okuda T. Tannins
of Stachyurus species. II. Praecoxins A, B, C and D,
four new hydrolysable tannins from Stachyurus
praecox leaves, Chem. Pharma. Bull., 1991; 39:
1689-1693.
7. Khan NA, Iqbal SA. Importance of Medicinal
Plants. Discovery Publishing House Pvt. Ltd. New
Delhi, First Edition., 2011.
8. Kiritikar KR, Basu BD. An I.C.S. Indian Medicinal
Plants. International Book Distributors, Dehradun,
India., 1995; 1: 371-372.
www.ejbps.com
165