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 Packialakshmi N. et al/ International Journal of Research in Pharmaceutical and Nano Sciences. 3(5), 2014, 380 - 396.

Research Article CODEN: IJRPJK ISSN: 2319 – 9563

International Journal of Research

in
Pharmaceutical and Nano Sciences
Journal homepage: www.ijrpns.com

STUDIES ON STRYCHNOS POTATORUM SEED AND SCREENING THE WATER

QUALITY ASSESSMENT OF DRINKING WATER

N. Packialakshmi*1, C. Suganya1, V. Guru1

*1
PG and Research Department of Microbiology, Jamal Mohamed College (Autonomous), Tiruchirappalli,
Tamilnadu, India.

ABSTRACT
This research was aimed to finding the efficiencies of powdered seeds of Strychnos potatorum natural water treatment
agents alternative to the use of synthetic chemicals. The optimum dosages and turbidities were observed to the alum and
Strychnos potatorum. The results obtained the Strychnos potatorum prove the plant can be used for the treatment of
turbidity in drinking water. Performance of Strychnos potatoram seed extract as primary coagulant and compared with
the performance of alum. S. potatoram seed extract is effective as a prime coagulant compared with alum, it produces
water with slightly higher residual turbidity and residual color, but the residual turbidity and residual color are within
the WHO drinking water guideline values for turbidity (5 NTU) and color (15 TCU). The effectiveness of Strychnos
potatoram in the removal of turbidity, total hardness, pH, and total dissolved solids (TDS) has been investigated.
Prelimainary phytochemical screening were carried out and also IR - Spectrum were analysed. Strychnos potatorum
seeds column compounds were tested for their antibacterial properties against some pathogenic gram positive and gram
negative bacteria. The growth of Proteus vulgaris, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus
epidermidis, Escherichia coli were significantly inhibited. The maximum zone of inhibition was found in Pseudomonas
aeruginosa, Klebsiella pneumoniae and Staphylococcus epidermidis.

KEYWORDS
Strychnos potatorum Seeds, MPN, Phytochemical, IR - Spectrum and Antibacterial activity.

INTRODUCTION
Author for correspondence: Water is used for several purposes by humans but
N. Packialakshmi,
the level of purity of the water being consumed is
PG and Research Department of Microbiology,
very crucial since it has a direct effect on health.
Jamal Mohamed College (Autonomous),
Safe drinking water should generally be free from
Tiruchirappalli, Tamil Nadu, India.
heavy metals, turbidity, organic compounds and
pathogen. Conventional treatments of water often
Email: [email protected].
include sedimentation, filtration and disinfection.
Among the coagulating agents used in water

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 Packialakshmi N. et al/ International Journal of Research in Pharmaceutical and Nano Sciences. 3(5), 2014, 380 - 396.

treatment, ferric sulphate or alum (aluminium plant and dried under shade. After drying, it was
sulphate) is some of the most widely used salts. powdered and used for our studies.
Aluminium is strongly neurotoxic and may be Sampling Sites
involved in the development of Alzheimer’s disease. The water samples were collected from different
It is well known fact that most of the chemical areas such as Kallanai, Kollidam, Pulivalam,
disinfectants used for antibacterial activity generate Thottiyam, Veerampatti, Sathanur, Keeranur,
various unwanted chemicals known as disinfection Uyyakondanthirumalai and Kajamalai representing
by products (DBPs) in water. There DBPs are the locations along North, South, West, East, North
associated with harmful effects on humans such as East, North West, South East, South West directions
hemolytic anemia, cancer risk, nervous system effect of the Trichy Taluk. However, the sampling
and liver effects. Addressing these problems calls locations include urban as well rural areas.
out for a tremendous amount of research to be Sample Collection
conducted to identify robust new methods of The samples were collected in the month of May
purifying water at lower cost and with less energy, 2014 and taken in pre- cleaned polyethylene bottles.
while at the same time minimizing the use of Physico- Chemical Analysis of Drinking Water
chemicals impact on the environment. We highlight The collected samples were analyzed for different
some of the natural herbals like Strychnos physico - chemical parameters such as pH,
potatorum(nirmali) used by humans throughout alkalinity, Calcium, Magnesium, Total solids, Total
history in treating drinking water1. dissolved solids and Total suspended solids.
S. potatorum(nirmali) is a moderate-sized tree found Preparation of Plant Extract
in Southern and central parts of India, Sri Lanka and The Strychnos potatorum plant seed were collected
Burma, used predominantly as a traditional and dried at room temperature for 2 - 3 days and
medicinal extract. Seeds of S. potatorum are used in further dried at 60° C. The dried seed were extracted
dysuria, polyuria, urolithiasis, also in epilepsy. The with aqueous. The extracts were filtered with the
seeds resemble those of nuxvomica but are non- Whattman filter paper and then dried by using rotary
poisonous. The ripe seeds are used for clearing evaporator. The filtrate was stored in screw cap
muddy water. Sanskrit writings from India reported bottle at -20° C for further use.
that the seeds were used to clarify turbid surface Colorimeter
water over 4000 years ago which indicated that they A colorimeter is a device used in colorimeter. In
were the first reported plant-based coagulant used scientific fields the word generally refers to the
for water treatment. The nuts of this species of device that measures the absorbance of
Strychnos are very largely used in some parts of particular wavelengths of light by a
India for clearing muddy water. The tree, which specific solution. This device is most commonly
grows to a very large size, produces a shining, black, used to determine the concentration of a
one-seeded berry2. The present study also used to known solute in a given solution by the application
treat the seed for water quality assessment. of the Beer-Lambert law, which states that the
concentration of a solute is proportional to the
MATERIALS AND METHODS absorbance3.
Plant Sample Collection
Strychnos potatorum seeds were collected from MPN METHODOLOGY
Velur near Viralimalai, Pudukottai district, Tamil To detect the coliform bacteria from the collected
Nadu India. The seed were identified by the Rapinat water samples. Coliforms can be identified from the
Herbarium, St. Josephs College, Tiruchirappalli, following three methods.
Tamilnadu, India. The seeds were separated from the

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Presumptive test Mayer’s Reagent

Three sets of test tubes (each contain three tubes and Mercuric chloride (1.358 g) was dissolved in 60 ml
one is control tube) were taken. 10 ml of double of distilled water, potassium iodine (5 g) was
strength lauryl phosphate tryptose broth is added dissolved in 10 ml of distilled water. The two
into first set of tubes. In the second and third set of solutions were making up to 30 ml of distilled water.
tubes add 10 ml of single strength of lauryl tryptose Test for Flavonoids
broth. Durhams tubes were inserted in all tubes Ferric Chloride test
without air bubbles inside the Durham’s tube. The 1 ml of extract was taken and a few drops of dilute
setup is sterilized. After cooling 10 ml of water ferric chloride solution were added. The color
sample were added into first set of tubes, 1 ml in changed to pale green or red brown color which
second set of tubes and 0.1 ml in third set of tubes. indicates the presence of flavonoids.
After incubation all the tubes were observed for the Test for Saponins
presence of gas production. The number of coli Foam test
forms in the water sample was calculated by 1 ml of extract was diluted separately with distilled
comparing the standard chart. water to 20 ml and shaken with graduated cylinder
Confirmed test for 15 minutes. Formation of air bubbles indicates
A loopful of culture from positive lauryl tryptose the presence of saponins.
broth was transferred to brilliant green lactose bile Test for carbohydrates
broth tubes. The tubes were incubated at 37° C for 24 Molisch’s test
hours. The tubes were examined for the presence of Small quantity of extract was dissolved separately in
gas in Durham’s tube within 48 hours, which 4 ml of distilled water and filtered, 2 ml of filtrate, 2
constitute a positive confirmed test. drops of alcoholic napthol solution are added. The
Completed test mixture is shaken well and 1 ml of concentrated
A loopful of culture from the positive tubes were sulphuric acid is added slowly along the sides of the
transferred to the Eosin methylene Blue agar test tube and allowed to stand. Formation of reddish
plates. The EMB agar plates were incubated at 37° C brown ring or violet ring indicates the presence of
for 24 hours. Following incubation the plates were carbohydrate.
examined5. (Dubey and Maheswari 2006). Test for tannins
Enumeration of microbes by spread plate Lead acetate test
technique To 5 ml of extract solution and 1 ml of lead acetate
Prepare plates with suitable nutrient medium place solution was added. Flocculant brown precipitate
0.1 ml of diluted sample in the center of the plate. indicates the presence of tannins.
Spread the inoculum over the medium by pushing Test for sterols
the glass spreader (L –Rod) backward and forward Libermannburchard reaction
while rotating the plate. Incubate the plates at A small amount of extract of sample and a few
suitable temperature6. crystal of sodium nitrate were taken in a dry test
tube and heated gently for a minute. It was cooled
PHYTOCHEMICAL ANALYSIS7 and added 0.5 ml of concentrated sulphuric acid.
Test for Alkaloids Orange color indicates the presence of sterols.
Mayer’s test Test for glycosides
To the little amount of extract were taken and add a A portion of the extract was hydrolysed with
few drops of Mayer’s reagent formation of hydrochloric acid for few hours on a water bath and
precipitate indicate the presence of alkaloids. the hydrolysate was subjected to legal’s test to detect
the presence of different glycosides.

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Legal’s test with 20 ml petroleum ether and 20 ml of 10 %

1 ml of sodium nitroprusside solution was added and aqueous sodium chloride solution. The separating
then it was made alkaline with sodium hydroxide funnel was shaken carefully and the lower layer was
solution. If the extract produced pink to red color, allowed to drain the beaker.
which indicates the presence of glycosides. Preparation of Column
Test for oil and fats A plug of cotton is placed to the bottom of the
Few drops of 0.5N alcoholic potassium hydroxide column so that silica and soil will not pass through
were added to small quantity of various extract the column. Slurry of silica was prepared and poured
along with a drop of phenolphthalein. The mixture into the column carefully. It is allowed to settle and
was heated on a water bath for 1-2 hours. Formation sand is added to the upper portion of the silica.
of soaps or particle neutralization of alkali indicates Loading a Sample
the presence of fixed oil and fats. The sample was added using a pasture’s pipette
Test for phenolic compounds carefully above the sand. The eluent is added on top
Few drops of extracts were taken separately in water of the sand. The mobile phase slowly flows down
tested for the presence of phenolic compounds with through the silica gel column by gravity leaving
dilute ferric chloride solution (5%) which gives behind zone of colour and a component was eluted
violet color. from column.
Test for protein and Aminoacids Microbial strains used
Biuret test Different microbial strains were used to evaluate the
A few drops of extract were taken in water and 1 ml antimicrobial effect of which two were gram
of 4% copper sulphate was added to it. Violet or positive bacterial strains (i.e) Staphylococcus
pink colour is conformed proteins are present. aureus, staphylococcus epidermidis, and three
Test for gums and mucilage strains were gram negative bacterial strains (i.e)
About 10 ml of the extract was added to 25 ml of Escherichia coli, Proteus vulgaris, Klebsiella
absolute alcohol with stirring and filtered. The pneumonia. The strains were obtained from Jamal
precipitate was dried in air and examined for its Mohamed College, Trichy, Tamil Nadu, India and
swelling properties and for the presence of maintained agar slants.
carbohydrates. Disc Diffusion Method
IR – spectrum analysis Disc diffusion method was carried out for
FTIR relies on the fact that the most molecules antibacterial susceptibility testing according to the
absorb light in the infra-red region of standard method to assess the presence of
electromagnetic spectrum. This absorption antibacterial activities of the plant extract
corresponds specifically to the bonds present in the (Ravikumar, 2011; Kim, 1999; Nusrat S, 2008) (8,9).
molecule. The frequency ranges are measured as Muller Hinton agar (MHA) plates were prepared.
wave numbers typically over the range 4000 - Overnight nutrient broth culture of test
400 cm-1. organisms were seeded over the MHA plates
Column chromatography using sterile cotton swab so as to make lawn
Column chromatography is used to purify liquids by culture. The discs which had been impregnated
separating an organic solvent from mixture of with aqueous extracts of leaf were placed on
solvent. the MHA with the control disc and subjected to
Preparation of seed extract antibacterial screening. The plates were then
The seed extract was prepared by grinding the incubated at 37° C for 18 to 24 hours
mixture in mortar and pestle containing 22 ml of depending on the species of bacteria used in this
acetone 3 ml of petroleum ether and calcium test. After the incubation, the plates were examined
carbonate. The pigments was filtered and mixed for inhibition zone.

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Chi – Square Test (x²) concentration is given in ( Table No.7). Best zone
In this study chi - square test (x²) was applied. The of inhibition was produced against Pseudomonas
purpose of chi – square test (x²) was to decide aeruginosa (23mm), and better zone of
whether the set of observed data ( Antibiogram inhibition against Klebsiella sp., (18 mm),Bacillus
of microorganisms) agrees with the standard subtilis(14 mm), Proteus vulgaris (14 mm),
antimicrobial disc susceptibility test (NCCLS, Escherichia coli (15 mm), and least was produced
2002). against Staphylococcus aureus (13 mm),
Staphylococcus epidermidis (12 mm), Maximum
RESULTS zone of inhibition were observed in
The present study deals with the treatment of Pseudomonas sp., and Klebsiella sp., and least
drinking water by using plant Strychnos potatorum zone of inhibition observed in Staphylococcus
seed powder. Drinking water contains the coliform aureus and Staphylococcus epidermidis
bacteria it was observed by using MPN method, (Figure No.11).
Eosin Methylene Blue agar plate confirmed the Seed column compound crystals were confirmed
coli forms (Figure No.5). Seed powder and in polarizing microscope (Figure No.10), The
chemical substance alum were added with the result of the antibacterial activity of column
drinking water separately and mixed well, separated compounds 64 µg concentration is
turbidity level will observed in colorimeter. given in (Table No.8). Best zone of inhibition
Initial and after adding (seed powder and alum) was produced against Pseudomonas aeruginosa
the OD values are note down, after that (32mm), Escherichia coli(24 mm), Klebsiella sp.,
colonies were observed by using Spread plate (24 mm), and better zone of inhibition against
method. Initial level too much of colonies were Proteus vulgaris (16 mm), Bacillus subtilis (16
present in the petriplates (Figure No.6), alum mm), Staphylococcus aureus (18 mm), and least
will inhibit the colonies within 30 minutes was produced against Staphylococcus epidermidis
(Figure No.7) and seed powder will inhibit the (14 mm), Maximum zone of inhibition were
organisms within 1 hour (Figure No.8). To observed in Pseudomonas sp., and Klebsiella sp.,
analyzed the Physical and chemical parameters of E.coli and least zone of inhibition observed in
drinking water before and after treating with Staphylococcus epidermidis (Figure No. 12). Seed
Strychnos potatorum seed (Table No.2 and 3). compound gives the maximum zone of
The present study shows the presence of inhibition, when compared with the seed crude
Strychnos potatorum using the organic solvent extract .
extraction technique the bioactive constituents
have been extracted and subjected to preliminary DISCUSSION
colour test to identify the nature of the In earlier studies Rajendran et al., (2013) shows
compounds such as Alkaloids, Reducing sugar, the Seeds of Strychnos potatorum(S. potatorum)
Phytosterol, phenolic compounds, tannins, protein and Moringa oleifera(M. oleifera) have shown
and amino and acids, absence of gums and promising result as the source of natural
mucilage, fixed oil and fats, were confirmed by coagulant in the clarification of turbid water.
suitable chemical test (Table No.4), Seed crude Direct filtration with S. potatorum seeds as
powder and column separated compounds having coagulant appeared effective in clarifying turbid
the functional groups, it was identified through water. This property is attributed due to the presence
IR – Spectrum (Table No.5 and 6) (Figure No.1 and of polyelectrolytes, proteins, lipids, carbohydrates
2). and alkaloids containing the –COOH and free -OH
The result of the antibacterial activity of surface groups in the seed. Among the other plant
Strychnos potatorum seed crude extract 64 µg materials investigated, seeds of M. oleifera were

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 Packialakshmi N. et al/ International Journal of Research in Pharmaceutical and Nano Sciences. 3(5), 2014, 380 - 396.

found to be one of the most effective sources of Pharmacognostic and phytochemical characteristics
primary coagulant for water treatment1. of leaves of an endangered medicinally important
In previous Vijayaraghavan and Sivakumar et tree species Strychnos potatorum13. In previous
al., studied the application of plant based study Packialakshmi et al., (2014) deals with the
coagulants for waste water treatment. Natural phytochemical characteristics of leaves and bark of
coagulants function by means of adsorption an endangered medicinally important tree species of
mechanism followed by charge neutralization or strychnos potatorum. Preliminary phytochemical
polymeric bridging effect. Frequently studied plant- screening revealed the presence of alkaloids,
based coagulants include Nirmali seeds (Strychnos flavonoids, sapiens, carbohydrates, tannins, sterols,
potatorum), Moringa oleifera, Tannin and Cactus. glycosides, oil and fats, phenolic compounds,
Utilization of these coagulants represents important protein and amino acids, gums and mucilage. The
progresss in sustainable environmental technology functional groups were identified through IR –
as they are renewable resources and their application spectrum 14.
is directly related to the improvement of quality of In earlier studies Mallikharjuna et al., (2009)
10
life for under developed communities . alkaloid fractions isolated from Strychnos
The earlier study Mallikharjuna et al.,(2007) potatorum seed were tested for their
studied the phytochemical screening of Strychnos antimicrobial properties against some pathogenic
potatorum seed sample and described along with gram positive, gram negative and acid – fast
physical and chemical compound such as bacteria and fungi. These fractions have shown
Alkaloids, Reducing sugar, Phytosterol, Fixed oil considerable antimicrobial activity against both
(11)
and fats, phenolic compounds, and tannins .In bacteria and fungi at the tested concentrations
earlier studies Venkatesh et al (2011) a phyto – (100 and 200 µg / ml)15. In previous study the
pharmacological review on Strychnos potatorum Packialakshmi et al., (2013) Strychnos potatorum
linn, this review will be helpful to create against some water borne pathogens, 62 µg
interest towards Strychnos potatorum and may be concentration the zone of inhibition occurred in
useful in developing new formulations with Salmonella sp., Klebsiella sp., Enterobacter sp.,
more therapeutic and economical value12. E.coli16.
The previous study was showed by Srikanth
Kagithoju. et al., (2013) to investigate the
Table No.1: Water samples collection from different places of Tiruchirappalli District
S.No Places of water samples collected Types of water sample
1 Kallanai – Vennaru River
2 Kallanai – Kollidam River
3 Thottiyam River
4 UyyakondanThirumalai Well
5 Kajamalai Well
6 Pulivalam Well
7 Kuruvikarankulam well
8 Gundur pond
9 Sathanur Pond
10 Veerampatti Pond

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Table No.2: Physical and chemical parameters of water samples before treating with Strychnos
potatorum seed
EC CO3 HCO3 Cl SO4 Ca Mg Na K Na2CO3
S.No pH -1
(dsm ) mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L
1 7.5 0.67 - 2.4 4.3 - 2.0 1.7 2.7 0.3 -
2 7.9 0.61 1.0 3.3 1.8 - 1.9 1.3 2.6 0.3 1.1
3 7.5 0.72 - 4.3 2.9 - 1.8 1.9 3.2 0.3 0.6
4 7.7 0.54 1.2 2.3 1.9 - 1.3 1.5 2.4 0.2 0.7
5 7.4 1.44 0.4 8.7 5.3 - 4.0 3.2 6.9 0.3 2.6
6 8.4 0.83 2.2 4.4 1.7 - 1.9 0.8 5.3 0.3 3.9
7 8.1 2.4 4.0 11.9 8.1 - 4.8 9.1 9.8 0.3 2.0
8 7.9 0.74 - 4.9 2.5 - 1.9 1.8 3.4 0.3 1.2
9 7.7 0.57 - 3.3 2.4 - 1.1 1.5 2.8 0.3 0.7
10 7.1 0.11 - 0.4 0.7 - 0.4 0.6 0.01 0.13 -

Table No.3: Physical and chemical parameters of water samples after treating with Strychnos potatorum
seed
EC CO3 HCO3 Cl SO4 Ca Mg Na K Na2 CO3
S.No pH
(dsm-1) mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L
1 5.1 0.63 - 4.6 1.7 - 2.2 3.0 0.72 0.40 -
2 4.7 1.17 - 8.0 3.7 - 2.9 3.4 4.3 1.11 1.7
3 5.2 0.90 - 5.1 4.0 - 1.9 2.4 3.91 0.85 0.8
4 4.8 0.64 - 4.2 2.3 - 1.5 1.9 2.51 0.52 0.8
5 5.1 1.52 - 10.1 5.2 - 4.1 3.4 6.88 0.85 2.6
6 4.9 0.91 - 7.2 2.0 - 1.8 1.4 5.59 0.39 4.0
7 5.7 1.45 - 8.8 5.7 - 3.4 2.4 8.01 0.66 3.0
8 4.5 0.78 - 5.2 2.6 - 1.8 1.4 4.24 0.42 2.0
9 4.7 0.75 - 2.6 4.9 - 1.6 1.3 4.18 0.34 0.3
10 5.0 0.36 - 2.1 1.5 - 0.9 1.8 0.71 0.21 0.6

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Table No.4: Phytochemical analysis of Strychnos potatorum seed

S.No Phytochemical Constituents Seed Extract
1 Alkaloids Positive
2 Flavonoids Positive
3 Saponins Positive
4 Carbohydrates Positive
5 Tannins Positive
6 Sterols Positive
7 Glycosides Positive
8 Oil and Fats Negative
9 Phenolic Compounds Positive
10 Protein and Amino acids Positive
11 Gums and mucilage Negative

Table No.5: Infrared spectrum analysis by Strychnos potatorum seed crude powder
S.No Peak value Stretching Interpretation
1 673.16 - Benzene
2 1089.78 C-C Stretching Secondary alcohols
3 1440.83 C-H Bonding CH2 Symmetric
4 1462.04 C-H Bonding Alkyl group
5 1525.69 C=C Stretching Alkenes
6 1558.48 C=C Stretching Alkenes
7 1639.49 C=O Stretching Amino acid
8 1651.00 C=O Stretching Amino acid
9 1687.71 C=O Stretching α, β- unsaturated, 6 membered ring
10 1705.07 C=O Stretching Aryl group
11 2854.65 C-H Stretching Alkyl group
12 2924.09 C-H Stretching Methyl group
13 3437.15 O-H Stretching Alcohol group

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Table No.6: Infrared spectrum analysis by Strychnos potatorum of seeds column separated compounds
S.No Peak value Stretching Interpretation
1 601.79 C-Cl Stretching Halogen
2 655.80 C-Cl Stretching Halogen
3 923.90 C-H Def Alkenes
4 1074.35 C-O Stretching Ether
5 1276.88 N=O Stretching Nitro compounds
6 1344.38 N=O Stretching Nitro compounds
7 1413.82 C=C Stretching Aromatic compou nds
8 1635.64 N=O Stretching Nitro compounds
9 2266.36 N-H Stretching Amino acids
10 2382.09 N-H Stretching Amino acids
11 2927.94 O-H Stretching Alcohols
12 3232.70 N-H Stretching Amide
13 3772.76 N-H Rocking Amines

Table No.7: Antibacterial activity of Strychnos potatorum seed crude extract

(Zone of inhibition in mm)
Seed crude extract
Bacterial strains
S.No Sample µg/ml Standard Observed
used X2 = Ʃ[(O-E)2]/E
value value
1 E. coli 21 15 1.714
2 Klebsiella sp., 21 18 0.727
3 P. vulgaris 21 14 2.909
4 Strychnospotatorum seed 64µg P. aeruginosa 21 23 0.190
5 S. aureus 21 13 3.047
6 S. epidermidis 21 12 3.857
7 Bacillus subtilis 21 14 2.909

Table valuex2 (0.05) = 3.841, Chi – square value significance at 5% level

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Table No.8: Antibacterial activity of Strychnos potatorum seed column extract

(Zone of inhibition in mm)
Seed column extract
S.No Sample µg/ml Bacterial strains Standard Observed X2 = Ʃ[(O-E)2]/E
used value value
1 E. coli 22 24 0.181
2 Klebsiella sp., 22 24 0.181
3 P. vulgaris 22 16 1.636
Strychnospotatorum
4 64µg P. aeruginosa 22 32 4.545
column compound
5 S. aureus 22 18 0.727
6 S. epidermidis 22 14 2.909
7 Bacillus subtilis 22 16 1.636
Table value x 2 (0.05) = 3.841, Chi – square value significance at 5% level

Figure No.1: Infrared spectrum analysis by Strychnos potatorum seed crude powder

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Figure No.2: Infrared spectrum analysis by Strychnos potatorum seed column separated compound

Figure No.3: Strychnos potatorum plant

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Figure No.4: Strychnos potatorum seed and seed powder

Figure No.5: Eosin Methylene Blue agar plate confirm the coli form bacteria

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Figure No.6: Water sample initial stage using spread plate method

Figure No.7: Water samples treated with S.potatorum seed powder

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Figure No.8: Water samples treated with alum

Figure No.9: Strychnos potatorum seed Column chromatography and S. potatorum seed dried crystals

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Figure No.10: Strychnos potatorum seed separated column compound crystals observed in Polarizing
Microscope (Size; 100µm / 161 pixels)

Figure No.11: Zone Inhibition formed by Strychnos potatorum seed crude extract

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Figure No.12: Zone Inhibition formed by strychnos potatorum seed column extract

CONCLUSION 3. "Colour." Encyclopædia Britannica:

This research study concludes that the Strychnos Encyclopedia Britannica Online, Encyclopedia
potatorum seed are very effective for drinking Britannica Inc., Web, 17 Nov, 2011.
water treatment and also it is very effective against 4. Kannan N. Laboratory manual in general
the pathogenic organisms. microbiology, Palani Paramount Publication,
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