The present study is the comparative analysis of phytochemical constituents and microbial load of an edible plant Alternanthera philoxeroides (Mart.) Griseb collected from unpolluted and polluted site. Preliminary phytochemical analysis was performed with acetone, aqueous, chloroform, ethanol and petroleum ether extracts (unpolluted and polluted site) of A philoxeroides that showed the presence of alkaloids, carbohydrates, saponins, phenols, flavonoids, aminoacids, diterpenes, tannin, terpenoids, protein, steroid, oxalate, coumarin and quinones. The ethanol extract showed higher number of phytochemical constituents when compared to the other extract of unpolluted site. The microbial load is also enumerated in the unpolluted and polluted site. In conclusion, phytochemical analysis revealed the presence of many phytoconstituents in ethanol extract and the microbial load is less in the unpolluted site when compared to the polluted site.

1. Effect of environmental pollution on the quality of an edible plant Alternanthera philoxeroides (Mart.) Griseb
IRJBB
Effect of environmental pollution on the quality of an
edible plant Alternanthera philoxeroides (Mart.) Griseb
U. Arul Pamila1*
and S. Karpagam2
1,2
Department of Botany, Queen Mary's College (A), Chennai, India.
The present study is the comparative analysis of phytochemical constituents and microbial load
of an edible plant Alternanthera philoxeroides (Mart.) Griseb collected from unpolluted and
polluted site. Preliminary phytochemical analysis was performed with acetone, aqueous,
chloroform, ethanol and petroleum ether extracts (unpolluted and polluted site) of A
philoxeroides that showed the presence of alkaloids, carbohydrates, saponins, phenols,
flavonoids, aminoacids, diterpenes, tannin, terpenoids, protein, steroid, oxalate, coumarin and
quinones. The ethanol extract showed higher number of phytochemical constituents when
compared to the other extract of unpolluted site. The microbial load is also enumerated in the
unpolluted and polluted site. In conclusion, phytochemical analysis revealed the presence of
many phytoconstituents in ethanol extract and the microbial load is less in the unpolluted site
when compared to the polluted site.
Keywords: Alternanthera philoxeroides, phytochemical constituents, microbial load.
INTRODUCTION
Medicinal plants have been a basic source of antibiotics
against a variety of diseases over the years. Herbal
plants are known to be the excellent stimulators of
immune system due to antioxidant properties of
phytochemical constituent. Herbal plants have been
recently popularized in modern medicine, since many
therapeutically important compounds are derived from
them (Ghosh et al., 2006. Herbal plant based drugs have
been in use against various infections (Ahmad et al.,
2009). In the past, medicinal plants was the first line of
treatment known to man and traditional medicinal
practice remain an important part of the primary
healthcare delivery system in most of the developing
world (Akerele., 1998). Phytochemicals are naturally
occurring in the medicinal plants in all the parts namely,
leaves, stem and roots that have defense mechanism
and protect from various diseases. Phytochemicals are of
primary and secondary compounds. Chlorophyll, proteins
and common sugars are included in primary constituents
and secondary compounds have terpenoids, alkaloids
and phenolic compounds (Wadood et al, 2013;
Krishnaiah et al,. 2007).
Plants accumulate various secondary metabolites
including alkaloids, glycosides, polyphenols etc. They are
effective in the treatment of infectious diseases with
minimal side effects that are often associated with
synthetic antimicrobials (Iwu, et. al., 1999). A good
number of drugs that are being used today are isolated
from plant sources and some from animals or minerals.
Alternanthera Forsskaal species are invasive aquatic
weeds posing a strong threat to agro-biodiversity in
several countries in the world. Weeds are a continuous
and ubiquitous threat to agricultural productivity (Riaz et
al., 2009).
*Corresponding author: U. Arul Pamila, Department of
Botany, Queen Mary's College (A), Chennai, India.
pamilastalin2004@gmail.com
International Research Journal of Biochemistry and Biotechnology
Vol. 4(1), pp. 061-067, June, 2017. © w w w .premierpublishers.org, ISSN: 2167-0438x
ResearchArticle

2. Effect of environmental pollution on the quality of an edible plant Alternanthera philoxeroides (Mart.) Griseb
Pamila and Karpagam 062
B
Figure 1A. A. philoxeroides produces papery w hite flow ers on stalks, Figure 1B. philoxeroides forming
floating Cooum River Maduravoyal, Chennai
Alternanthera philoxeroides (Mart.) Griseb
(Amaranthaceae) is commonly known as alligator weed
and joyweeds (Fig. 1) that can be found in many parts of
the world, infesting rivers, lakes, ponds and irrigation
canals, as well as many terrestrial habitats. It is an
emergent stoloniferous perennial herb. The leaves are
dark green, elliptic, glabrous and opposite, 3.5-7.1 cm
long and 0.5-2 cm wide. Mature aquatic plants have
hollow stems up to 10 m long that form thick interwoven
mats throughout the water body and emerge up to 20 cm
out of the water when the plant flowers. Flowers are
white, inflorescences terminal and axillary, 1.4-1.7 cm in
diameter, on a short stalk. In the native range the species
is known to set seed. In much of the invasive range seed
production is not observed (Bassett, 2006).
Habitats: Although more suited to aquatic and riparian
habitats, where the species forms dense mats in shallow
slow-moving water bodies, it is also a vigorous colonizer
of terrestrial habitats where the extensive (up to 2 m)
deep rhizome system can sustain the population
throughout extended dry periods. Often, it grows at the
interface between the aquatic and terrestrial environment.
Spread is predominantly vegetative, from axillary buds at
each node in the warm summer months. (Julien and
Bourne, 1988) list a number of habitats which will sustain
populations of the species, including, but not exclusive
too, freshwater habitats, coastal areas, managed
terrestrial habitats including cultivated agricultural land,
disturbed areas and urban habitats. In addition, natural
and semi-natural habitats are prone to invasion, including
forests, riverbanks and wetlands (Clements et al., 2014).
It is an emergent, semi-aquatic plant which originated in
South America, introduced range includes New Zealand,
Australia, Asia, and more recently parts of Europe
(Julien, 1995); A. philoxeroides is found throughout India,
including Assam, Bihar, West Bengal, Tripura, Manipur,
Andhra Pradesh, Karnataka, Maharashtra, Delhi and the
state of Punjab (Pramod et al., 2008).
A philoxeroides plays a vital role in human health care.
All parts of plants are used in traditional systems of
medicine for the treatment of numerous human ailments.
They were reported to treat bacterial, fungal and viral
diseases. Young tender shoots and leaves of
A.philoxeroides are eaten as a vegetable in Southeast
Asia (Scher Federal., 2009). The plant is used as leafy
vegetable. It contains an immense variety of bioactive
compounds such as antimicrobials, antioxidants,
phytochemicals, essential fatty acids and dietary fibres
(Gupta and Prakash., 2008). Due to its dietary
importance, many scientific studies have been carried out
on the nutritive values of herbal plant of A. philoxeroides
(Gayathri et al., 2006). Therefore, the present work is
aimed to compare and explore the microbial load and
phytochemical analysis of acetone, aqueous, chloroform,
ethanol and petroleum ether extract of Aphiloxeroides
collected from unpolluted and polluted sites Pechiparai of
Kanyakumari District, Tamil Nadu and Cooum River,
Maduravoyal (Fig. 2).
MATERIALS AND METHODS
Collection and authentication of plant materials
The A. philoxeroides specimens were collected from two
different sites Pechiparai of Kanyakumari District, Tamil
Nadu and Cooum River Maduravoyal, Chennai. The
collected plants were identified in the Department of
Botany, Queen Mary's College and confirmed by Prof. P.
Jayaraman, Director, Plant Anatomy Research Centre
(PARC) Chennai.
Plant material
Fresh plants were washed thoroughly three to four times
with running tap water then finally with sterile water
followed by shade drying at room temperature for 20-30

3. Effect of environmental pollution on the quality of an edible plant Alternanthera philoxeroides (Mart.) Griseb
Int. Res. J. Biochem. Biotechnol. 063
Figure 2. Location map of study area
days and powdered by using an electric blender and
stored in airtight container.
Preparation of extract
Each sample of 10g were taken and soaked for 24h in
30ml of aqueous, acetone, chloroform ethanol, and
petroleum ether separately. The extracts were filtered
using Whatman filter paper No. 1, evaporated to dryness
and re-dissolved in DMSO (Dimethyl Sulphoxide). The
extracts were preserved in airtight container and kept at
4-5
0
C for further use.
Enumeration of microbial load from fresh plants
Freshly collected plant twig of 10 cm was washed
in 100ml of sterile water; the twig was placed on sterile
blotting paper to remove excess water and again washed
in 100ml of sterile water and repeated again for third
wash. The three sterile waters were serially diluted and
plated on NA plates separately.
The pour plate method was used to cultivate
serially diluted portions of the medicinal plant samples
under investigation. Enumeration was carried out on
nutrient agar. Triplicate plates of appropriate dilutions
were prepared. The NA plates were incubated at 37
0
C
for 24h for bacterial growth. The developed microbial
colonies were counted and tabulated.
Enumeration of microbial load from powdered plant
samples
The plant sample (0.1g) was added to 20ml of lukewarm
NA media, mixed thoroughly and poured in Petriplates,
incubated it for 24h and observed for bacterial colonies.
The developed microbial colonies were counted.
The plant sample (0.1g) was suspended in 1ml sterile
water shaken well and serially diluted and 0.1 ml was
plated on NA plates and incubated. After 24h, the
developed microbial colonies were counted and
tabulated.
Phytochemical Analysis
The plants have primary and secondary metabolites
which can be used for medicinal and other uses. There
is a need to analyze the plants for such phytochemical
screening. Phytochemical screening was carried out by
using the standard protocols as described by Harborne
(1973). The alkaloids are determined by Wagner’s Test
(Tiwari et al., 2011); carbohydrates by Benedict’s Test;
saponin by Foam Test phenol by Ferric Chloride Test;
flavonoids by Lead Acetate Test; diterpenes by Copper
Acetate Test; aminoacids by Ninhydrin Test and Tannins
by Ferric Chloride Test; terpenoids by Salkowski's Test,
proteins by Biuret Test (Khanam et al., 2014) and oxalate
by Ethanoic acid glacial (Ugochukwu et al., 2013).
Further detection of steroids was carried out by Harborne
(1973); detection of coumarin was done by Mace method
(Mace, 1963) and quinone by conc. H2SO4.
Xanthoproteins by conc. HNO3 and NH3 Test, carboxylic
acid by effervescence test (Kumar et al., 2013) cardiac
glycosides by Kellerkillani synthesis (Misra et al., 2011)
anthocyanin by HCl and NH3, leucoanthocyanin by
isoamyl alcohol (Ashvin Godghate et al., 2012); and
glycosides by Modified Borntrager's Test (Kokate et al.,
2006).

4. Effect of environmental pollution on the quality of an edible plant Alternanthera philoxeroides (Mart.) Griseb
Pamila and Karpagam 064
Table 1: Phytochemical content of A. philoxeroides collected from unpolluted and polluted site.
Name of the test
Ethanol Chloroform Acetone Petroleum ether Aqueous
UP P UP P UP P UP P UP P
Alkaloids + + - + - + - + + +
Carbohydrates + - + - - - + - - -
Saponins + - + - - - + - + -
Phenols + - - - - - - - + -
Flavonoids + + - - + - - - + -
Aminoacids + + - - - - - - + +
Diterpenes + + + + + + + - + +
Tannins + - - - - - - - - -
Terpenoides + + - - + - + + + +
Protein + + + - - - + + + +
Steroid + + + - - - + - - +
Oxalate + + + - + - + - - -
Cardiac glycosides - - - - - - + - - -
Carboxylic acid - - - - - - - - - +
Xanthoprotein - - + - - - + - - -
Coumarin + - + - + - + - + -
Quinones + + - - + - - + + -
Glycosides - - - - - + - - + +
Note: "+" indicates present and "-" indicates absent. Anthocyanin and Leucoanthocyanin absent; UP: Unpolluted site
plant and P-polluted site plant.
Table 2: Analysis of microbial load of unpolluted and polluted site collected from fresh plant A. philoxeroides
Washing a twig of
plant
Unpolluted site
(Fresh plant
twig)
Polluted site
(Fresh plant
twig)
I wash 57 x 109 Uncountable
II wash 34 x 109 187 x 109
III wash 13 x 109 69 x 109
RESULTS AND DISCUSSION
The phytochemical analysis was carried out from the
dried (polluted site and unpolluted site) plant powder
using aqueous, acetone, chloroform, ethanol, and
petroleum ether, which showed the presence of many
bioactive compounds in the plant in Table 1. Twenty
different phytochemical tests were carried out for the five
different extracts. Ethanol extracts showed the presence
of fourteen major phytoconstituents in unpolluted site
plant and nine major phytoconstituents in polluted site
plant. The preliminary phytochemical analysis showed
the presence of alkaloids, carbohydrates, saponins,
phenols, flavonoids, aminoacids, diterpenes, tannin,
terpenoids, protein, steroid, oxalate, coumarin and
quinone in the ethanol extract (unpolluted site plant). The
presence of these metabolites suggests great potential
for the plant as a source of useful phytomedicines (Kunle
et al., 2003). Antibiotic activities are common for
alkaloids some are even used as antiseptics in medicines
(Cordell 1983).
The preliminary phytochemical analysis showed primary
and secondary metabolites such as alkaloids,
carbohydrates, saponins, phenols, flavonoids, diterpenes,
tannin, terpenoids, steroid, oxalate, anthocyanin,
leucoanthocyanin, Xanthoprotein, coumarin and
glycosides in the ethanol extract of A bettzickiana (Pamila
and Karpagam., 2017). The presence of carbohydrates,
aminoacids, proteins, cardiac glycosides, alkaloids,
steroids, flavonoids, total phenolics and tannin contents
were reported in A.philoxeroides (Fang et al., 2006). The
medicinal value of plants depends on the presence of
phytoconstituents. The chloroform extract showed the
presence in minimal amount of phytoconstituents. The
results showed the presence of some secondary
metabolites which proves that the plant is of great
medicinal values.
The results of the microbial load of fresh plant
A.philoxeroides collected from different location namely
unpolluted and polluted site are presented in Table 2. In

5. Effect of environmental pollution on the quality of an edible plant Alternanthera philoxeroides (Mart.) Griseb
Int. Res. J. Biochem. Biotechnol. 065
(A)
(a) (b) (c)
(B)
(d) (e) (f)
Plate 1: Photographs of microbial load of polluted (A) and unpolluted (B) site collected from fresh plant
A.philoxeroides; (A). Fresh plant tw ig polluted (a) I w ash, (b) II w ash, (c) III w ash; (B) Fresh plant tw ig
unpolluted (e) I w ash, (f) II w ash, (f) III w ash.
Table 3: Analysis of microbial load of unpolluted and polluted site dry plant pow er of A.philoxeroides
Plant dry powder (0.1g powder diluted 1ml of
sterile water)
Unpolluted site Polluted site
Dilution 7 15 x 10
7
36 x 10
7
Dilution 8 10 x 10
8
29 x 10
8
Dilution 9 4 x 10
9
17 x 10
9
(C)
(g) (h) (i)
(D)
(j) (k) (l)
Plate 2: Photographs of microbial load of polluted (C) and unpolluted (D) site dry plant pow der sample C-
polluted dry plant pow der: (g) 7th
dilution, (h) 8th
dilution, (i) 9th
dilution; D-Unpolluted dry plant pow der: (j) 7th
dilution, (k) 8th
dilution, (l) 9th
dilution.

6. Effect of environmental pollution on the quality of an edible plant Alternanthera philoxeroides (Mart.) Griseb
Pamila and Karpagam 066
the polluted site plant, the microbial load is uncountable
in first wash; the microbial load is 187x10
9
cfu/g in
second wash; and the microbial load is 69x10
9
cfu/g in
third wash. In unpolluted site plant, the microbial load is
57x10
9
cfu/g in first wash, 34x10
9
cfu/g in second wash,
and 13x10
9
cfu/g in third wash,
The results of the microbial load of dry powdered sample
A. philoxeroides are presented in Table 3. The result of
the microbial load of the polluted site dry plant powder
sample counted 42 colonies and unpolluted site counted
21 colonies. The result of the microbial load of polluted
site dry plant powder developed colonies 36x10
7
cfu/g,
29x10
8
, and 17x10
9
, unpolluted site dry plant powder
developed colonies 15x10
7
cfu/g, 10x10
8
, and 4x10
9
cfu/22-g. Contamination by microorganisms is
dispensing hygiene is not good, contamination may be
even worse (Obuekwe et al., 1998). Inadvertent
contamination by microbial or chemical agents during
processing could also caused deterioration, thereby
compromising safety and quality, and rendering the
medicinal plant material less effective and possibly affect
the consumer (W.H.O., 2003). Thus, the result of the
microbial load is higher amount in the polluted site when
compared to the unpolluted site and also the microbial
count is more in the fresh plant when compared to the dry
plant powder.
CONCLUSION
These simple reliable standards will be useful to use
herbs as a home remedy. Also the manufacturers can
utilize them for identification and selection of the raw
material for drug production. The microbial load findings
from this study emphasized the need for constant quality
assessment of herbal drugs on sale in order to ensure
the production of therapeutic products suitable for human
consumption.
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Accepted 25 April, 2017.
Citation: Pamila UA, Karpagam S (2017). Effect of
environmental pollution on the quality of an edible plant
Alternanthera philoxeroides (Mart.) Griseb. International
Research Journal of Biochemistry and Biotechnology,
4(1): 061-067.
Copyright: © 2017 Pamila and Karpagam. This is an
open-access article distributed under the terms of the
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unrestricted use, distribution, and reproduction in any
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