Pharmacognostic Standardization of an Ethnomedicinal Aquatic Herb, Monochoria hastata (L.) Solms for its Antibacterial Potentiality

Abstract

Objectives: To evaluate antibacterial potentiality, pharmacognostic characteristics and quality control parameters including heavy metals, like lead (Pb) and arsenic (As) accumulation in the aerial parts of an aquatic herb, Monochoria hastata (L.) Solms. Methods: Antibacterial assay was done by agar well diffusion method. Pharmacognostic studies like morpho-anatomical and physicochemical analyses were carried out for organoleptic, microscopic and macroscopic evaluations of living aerial parts, and powder microscopy, fluorescence, proximate and elemental analysis of the dried powder sample. Elements and heavy metals contents were determined by carbon, hydrogen, nitrogen, sulphur / oxygen (CHNS/O) analyzer and inductively coupled plasma mass spectrometry (ICP-MS), respectively. Results: M. hastata (L.) Solms aerial parts showed antibacterial activity against gastrointestinal and topical pathogens. It exhibited an amphistomatic and hydromorphic anatomical characters. The distinguishing features were the presence of stomata in upper and lower epidermis, broad air chambers, cuticle, collateral vascular bundles, sclereidal fibres, pitted tracheids, pitted vessels, calcium oxalate crystals and annular vessels in leaf. The powder sample contained very less amount of acid insoluble ash than water soluble ash and complete absence of foreign organic matter. Carbon, hydrogen, nitrogen and sulfur (CHNS) ratio was 33:6:5:1 and lead content was lesser than the recommended upper limit though the arsenic content was higher than the permissible upper limit. Conclusions: Though the plant has traditionally been used as a potent Ethnomedicinal herb to cure boils, gastritis, hepatopathy and as laxative, but no such evaluation of pharmacognostic identity and quality parameters have been done so far. This is the first report on its pharmacognostic characters and quality control issues like heavy metal accumulation and physicochemical parameters for future use as powder drug.

Full text

Pharmacognosy Journal, Vol 10, Issue 3, May-Jun, 2018 533
Pharmacogn J. 2018; 10(3):533-540.
A Multifaceted Journal in the eld of Natural Products and Pharmacognosy
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Original Article
INTRODUCTION
Indigenous people use medicinal plants either as original
or as semi-synthetic herbal preparations in various
ailments without side eects.1 Medicinal plants are
considered as safer and cost eective also.2 70% of
Indian population use herbs as remedy based on their
traditional and indigenous knowledge. Indian Systems
of Medicine (ISM) including Ayurvedic, Unani, Siddha,
Homoeopathy, Naturopathy and tribal medicines use
about 400 plants. Herbs are the potent source of several
novel modern drugs. Pharmaceutical sector in India
use 280 medicinal plant species.3 Hydrophytes or
plants grown on aquatic, wet and marshy lands also
have medicinal uses like the plants grown in terrestrial
habitat.4 India is a country having a vast ethnic diversity
and is rich in dierent indigenous knowledge system.
Many ethnic communities inhabit in Malda
(24º 40′ 20″ N and 25º 32′ 08″ N latitude and 87º 45′
50″ E and 88º 28′ 10″ E longitude), a district situated
in West Bengal province of this nation where ‘Kaviraaj’,
Pharmacognostic Standardization of an Ethnomedicinal
Aquatic Herb, Monochoria hastata (L.) Solms for its Antibacterial
Potentiality
ABSTRACT
Objectives: To evaluate antibacterial potentiality, pharmacognostic characteristics and quality
control parameters including heavy metals, like lead (Pb) and arsenic (As) accumulation in the
aerial parts of an aquatic herb, Monochoria hastata (L.) Solms. Methods: Antibacterial assay
was done by agar well diffusion method. Pharmacognostic studies like morpho-anatomical
and physicochemical analyses were carried out for organoleptic, microscopic and macroscopic
evaluations of living aerial parts, and powder microscopy, uorescence, proximate and
elemental analysis of the dried powder sample. Elements and heavy metals contents were
determined by carbon, hydrogen, nitrogen, sulphur / oxygen (CHNS/O) analyzer and inductively
coupled plasma mass spectrometry (ICP-MS), respectively. Results: M. hastata (L.) Solms
aerial parts showed antibacterial activity against gastrointestinal and topical pathogens. It
exhibited an amphistomatic and hydromorphic anatomical characters. The distinguishing
features were the presence of stomata in upper and lower epidermis, broad air chambers,
cuticle, collateral vascular bundles, sclereidal bres, pitted tracheids, pitted vessels, calcium
oxalate crystals and annular vessels in leaf. The powder sample contained very less amount
of acid insoluble ash than water soluble ash and complete absence of foreign organic matter.
Carbon, hydrogen, nitrogen and sulfur (CHNS) ratio was 33:6:5:1 and lead content was lesser
than the recommended upper limit though the arsenic content was higher than the permissible
upper limit. Conclusions: Though the plant has traditionally been used as a potent Ethno-
medicinal herb to cure boils, gastritis, hepatopathy and as laxative, but no such evaluation of
pharmacognostic identity and quality parameters have been done so far. This is the rst report
on its pharmacognostic characters and quality control issues like heavy metal accumulation
and physicochemical parameters for future use as powder drug.
Key words: Fluorescence characteristics, Heavy metal accumulation, M hastata (L.) solms,
Pharmacognostic standardization, Traditional medicinal plants.
Debabrata Misra1, Manab Mandal1, Narendra Nath Ghosh2, Vivekananda Mandal1*
Debabrata Misra1, Manab
Mandal1, Narendra Nath
Ghosh2, Vivekananda
Mandal1*
1Plant and Microbial Physiology and
Biochemistry Laboratory, Department
of Botany, University of Gour Banga,
Malda, West Bengal, INDIA.
2Department of Chemistry, University
of Gour Banga, Malda, West Bengal,
INDIA.
Correspondence
Vivekananda Mandal
Department of Botany, University of
Gour Banga, Malda, West Bengal, INDIA.
Phone no: +91 9679008986
E-mail: mandalvivek@gmail.com
History
• Submission Date: 18-12-2017;
• Review completed: 23-12-2017;
• Accepted Date: 05-02-2018
DOI : 10.5530/pj.2018.3.87
Article Available online
http://www.phcogj.com/v10/i3
Copyright
© 2018 Phcog.Net. This is an open-
access article distributed under the terms
of the Creative Commons Attribution 4.0
International license.
Cite this article: Misra D, Mandal M, Ghosh NN, Mandal V. Pharmacognostic Standardization of
an Ethnomedicinal Aquatic Herb, Monochoria hastata (L.) Solms for its Antibacterial Potentiality.
Pharmacog J. 2018;10(3):533-40.
‘Vaidya’, ‘Ojha’, ‘Jan Guru’ and aged knowledgeable
persons of dierent ethnic communities living in
Malda are the Ethno-medicine practitioners (Figure 1).
ey have information regarding the use of dierent
medicinal plants curing several ailments.5
ere are many pertinent literatures which support
the Ethno-medicinal use of herbs in various diseases
and ailments. Some of the published works done so
far on the traditional uses of edible and medicinal
plants in Malda district are mentioned chronologically
in the Table 1.
Monochoria hastata (L.) Solms is one of the potent
Ethno-medicinal herbs used in India. Root stock
of this plant increases vitality.12 Young shoots
are used as green leafy vegetables and leaf juice is
applied to cure boils by Bodo, Koch-Rajbongshi
and Rangia tribes.13 Root stock and leaves of M.
hastata (L.) Solms are also used in gastropathy,

Misra et al.: Pharmacognostic Standardization of Monochoria hastata (L.) Solms
534 Pharmacognosy Journal, Vol 10, Issue 3, May-Jun, 2018
hepatopathy and anti-lipooxygenase (LOX) activity.14 e leaves of
the herb are laxative and its paste is served to
cattle with diarrhoea as a tonic.15 Hence, this herb might have some anti-
enteric ecacy against some gastrointestinal microora. In West Bengal,
this aquatic plant is used as medicinal herb and fodder.16 In Malda, this
plant species is dominant as emergent hydrophytes in the water bodies
such as Piasbari nursery pond, Sagardighi, Asokpally pond, Rajgunge,
pond near Ramkeli.17
e identity, quality, purity and safety of an Ethno-medicinal herb to be
used as powder drugs, is an important concern due to conventionally
used immediate natural sources may contain some hazardous contami-
nants including heavy metals, toxic substances and foreign materials. It
can be achieved by a stepwise standardization process which includes
pharmacognostic studies like organoleptic, macroscopic as well as
microscopic and eco-physiological features and also the evaluation of
physicochemical parameters, viz. moisture content, ash values, uorescence
analysis, as well as by toxicological investigation, viz. heavy metals and
foreign matter contents etc.18 Organoleptic or physical properties like
colour, odour, size, shape, taste etc.; microscopic and macroscopic char-
acters are very important to identify a plant as a crude drug source.19
Estimation of proximates such as ash values, loss on drying, presence of
foreign organic matter etc. in herbal material helps us to justify its safety
and ecacy and to ensure reproducible quality of it as crude powder
drug.20 Ash value gives an indication of its purity level in presence of
various impurities like carbonates, oxalates and silicates.21 Loss on drying
can be estimated by quantifying the amount of moisture present in the
plant sample. It is very much noteworthy regarding its storability as
moisture gives suitable growth condition to microorganisms contamina-
tion and spoilage.22 A medicinal plant material should be entirely free
from any visible sign of contamination by any organic foreign materials
such as moulds or insects, and other animal contamination, including
animal excreta as they may produce toxic substances like aatoxins.23
Fluorescence analysis is also an important tool for the screening of some
compounds which do not uoresce themselves but when they are treated
with some reagents and solvents are converted into uorescent derivatives.24
Medicinal properties of a plant directly or indirectly depend on the
elemental concentrations of that plant. Medicinal plants contain major
elements which are widely used as precursors of dierent metabolite
biosynthesis pathways. Elemental concentrations of medicinal plants
regulate the curative ability of certain diseases. Carbon and hydrogen
are the major elements commonly present in all organic molecules while
nitrogen and sulphur is present in some special organic compounds like
alkaloids, glucosinolates, camalexin, defensins etc.25 Hence, it is important
to know the elemental concentration in medicinal plants which gives
indications on what kind of organic metabolites are present in the plant
sample.26 Evaluation of CHNS ratio is also important as it may act as a
purity index. is ratio is aected when there is any adulterant in the
crude drug sample.
Water hyacinth (Eichhornia crassipes) and other aquatic plants have
been reported to uptake arsenic of dierent concentrations and thereby
reduce arsenic pollution.27 Investigation of heavy metals accumulation
in frequently utilized medicinal plants collected from environmentally
diverse locations of north western India proved that consumption of raw
medicinal herbs grown in toxic heavy metal sites caused serious conse-
quences on human health.28 Arsenic (As) contaminated drinking water
beyond the permissible limit has deleterious eects on human health viz.
cardiovascular problem, gastrointestinal disorders, haematological ailments,
neurological eects, skin diseases etc. Higher levels of lead (Pb) are
carcinogenic and eects on the central nervous system and memory
(dyslexia).29 Hence, checking the level of heavy metal contamination
in herbal plant material, specially the aquatic one is an important con-
cern before its medicinal use. International agencies like World Health
Organization (WHO), Food and Agriculture Organization (FAO) has
suggested the analysis of absorption, elimination and toxic proles of
Table 1: Published works on the traditional uses of edible and medicinal plants of Malda district.
Title of the work Name of the author (s) with year of
publication
Inventory of some ethno-medicinal plants in wetlands areas in Maldah district of West Bengal Chowdhury and Das (2009)6
Wild edible plants consumed by local communities of Maldah district of West Bengal, India Chowdhury and Mukherjee (2012)7
Some less known plants from Malda district of West Bengal used for the treatment of arthritis, rheumatism and
gout Mitra and Mukherjee (2013)8
Indigenous knowledge of plants in local healthcare management practices by tribal people of Malda district,
India Saha et al. (2014)9
Ethnobotany of Chanchal Block of Malda District of West Bengal (India): plants used in local healthcare Saha et al. (2014)10
Ethnobotany, traditional knowledge and socioeconomic importance of native drink among the Oraon tribe of
Malda district in India Saha et al. (2015)11
Figure 1: Ethnic communities residing in dierent development blocks
of Malda district of West Bengal. (Source: District Human Development
Report : Malda, Development and Planning Department, Government of
West Bengal (2007).

Misra et al.: Pharmacognostic Standardization of Monochoria hastata (L.) Solms
Pharmacognosy Journal, Vol 10, Issue 3, May-Jun, 2018 535
antibacterial assay. Overnight culture of each bacterial strain grown in
broth was diluted to 0.5 McFarland turbidity standard for inoculation
and seeded on Mueller Hinton agar (MHA) (HiMedia M173-500G)
plates by using sterilized swabs. e medium was allowed to dry under
laminar air ow and wells were cut by using sterilized corn borer
(7 mm diameter). 20 µl each of the concentrated extracts and fraction
was placed into the agar wells. 3µl Ciprooxacin antibiotic (50µg/ml)
was used as positive control and respective solvents as negative control.
Plates were incubated at 37°C for 24 h and diameter of growth inhibition
zones were determined.31
Organoleptic Evaluation
Various sensory parameters like color, odor, appearance, texture and
taste of the plant material had been studied as per standard procedure.32
Macroscopic and Microscopic Evaluation
Dierent characteristic features of leaf lamina and petiole of M. hastata
(L.) Solms like size, shape, margin, surface, venation pattern etc. including
the type of stem and nature of inorescence were morphologically studied.33
All the slides prepared for the leaf and powder microscopy were visual-
ized under a compound microscope (Magnus MLX) in 40X objective and
photographs were taken using the camera, Magnus (MagCam DC10).
Leaf and Powder Microscopy
In this study, fresh leaf was dipped in distilled water and transverse
sections (T.S.) were cut manually. in sections were mounted on glass
slide with glycerine without any staining reagent. Various histological
and eco-physiological characters, such as cell composition, adaptive
structures and histochemical features were qualitatively studied under
microscope and photomicrographs were taken. For powder microscopy,
shade dried leaves were nely powdered in a mixer-grinder machine and
studied under the microscope. A pinch of the powder was placed in the
water taken in a watch glass and remained undisturbed for 2 h. 1drop of
it was taken on a glass slide and stained with one drop of 1.0 % w/v
safranin in 70% ethanol. Aer 5 min it was washed two times with
distilled water and stained with 0.1% w/v light green in 90% ethanol
for 5 min. Aer washing two times with distilled water the slide was
covered with cover slip and examined under microscope. Dierent cell
components i.e. vessels, bres, cortex cells, calcium oxalate crystals were
studied.34
Biochemical test and Microscopy for Calcium Oxalate
Crystals
e presence of calcium oxalate crystals in the leaf tissues was conrmed
by the chemical method as described by.35 e leaf powder sample was
treated with 2N acetic acid for 15 min to remove phosphate and carbonate.
ese were then treated with 1% silver nitrate in 15% hydrogen peroxide
(1:1, v/v) for 15 min at 22°C. en the treated sample was washed in
distilled water and counter stained with 2% safranin for 3 min and
observed under compound microscope.
Determination of Extractive values
e powder sample was taken in conical asks and soaked with various
solvents like absolute n-hexane, diethyl ether, chloroform, dichloro-
methane, ethyl acetate, methanol, 90% methanol in water, 50% methanol
in water and water separately and capped. e capped asks were then
agitated in an orbital shaker at 100 rpm for 24 h. Aer obtaining the
ltered extract, it was then transferred into weighed Petri plates and
concentrated to dryness by keeping ltrates for complete evaporation
of solvent. e extractive value in percentage was calculated by using
following formula:
heavy metals in the herbal medicines and also put forwarded the other
relevant necessary chemical, biological, and environmental analysis as a
critical issue in their guidelines.30 ough very little is known regarding
the potential inuences of heavy metals on pharmacological activities
of plant derived natural drugs. Yet, the heavy metal content analysis is
necessary as a safety measure.
e present investigation was dealt with the study of antibacterial
bioassay guided organoleptic and morpho-anatomical studies, analysis of
orescence, estimation of proximate contents, determination of CHNS
ratio, levels of arsenic and lead heavy metals present in the leaf of the
aquatic medicinal herb, M. hastata (L.) Solms. is kind of study helps
to channelize indigenous knowledge of herbal use and contributes to
improve health care delivery system by alternative and complementary
medicines in today’s pluralistic society.
MATERIALS AND METHODS
Microorganisms and Chemicals
Gram ‘+’ bacterial strains of Staphylococcus aureus MTCC 96 (S. aureus)
and Streptococcus mutans MTCC 497 (S. mutans), and Gram ‘–’ strain of
Escherichia coli MTCC 571 (E. coli) were purchased from the Microbial
Type Culture Collection and Gene Bank (MTCC), Chandigarh, India. It
is an aliate member of the World Federation for Culture Collections
(WFCC) and is registered with the World Data Centre for Microorganisms
(WDCM). All the chemicals including n-Hexane, ethyl acetate, methanol,
formalin, absolute alcohol, safranin, fast green, acetic acid, canada
balsam, H2SO4, NaOH, KOH, H2SO4, HNO3, HCl, Na2HPO4, FeCl3, distilled
water, aniline, silver nitrate, hydrogen peroxide, acetic acid, silica gel,
light green, safranin etc. were of analytical grade and were procured
either from Merck Limited India or from Sisco Research Laboratories,
Mumbai, India.
Collection and Authentication of Plant Materials
Fresh leaves of M. hastata (L.) Solms were collected from Jalalpur
wetland of Kaliachak-I Development Block of Malda district in West
Bengal and was identied by Dr. R. Gogoi, Scientist-D, Central National
Herbarium of Botanical Survey of India, Howrah-711 103, West Bengal
(India) with the specimen voucher no. UGB/DM/01.
Processing of Plant Sample, Extraction and Partial
Purication
Collected plant materials were washed thoroughly under running tap
water, rinsed in distilled water and dried in hot air chamber at 50°C for
one week. Dried leaves were uniformly grinded using a mixer-grinder
machine and the leaf powder was stored at 4°C in an air tight container.
Fiy g of leaf powder soaked in 500 ml absolute ethyl acetate and 50%
methanol in water were separately extracted using a soxhlet apparatus
at 40°C for 48 hrs. e extracts were ltered through Whatman No. 1
lter papers and made pigment-free by passing through the activated
charcoal columns. e de-pigmented ltrates were collected and
concentrated using a rotary vacuum evaporator (Supert, R-150, Mumbai,
India). e concentrated extract was stored at 4 °C until use for antibac-
terial activity test. e ethyl acetate extract was further puried using
silica gel (100-200 mesh) column as the stationary phase, and n-Hexane
and ethyl acetate at a ratio of 1:1 (v/v) as the mobile phase. e puried
fraction was air dried and kept in an Eppendorf tube for further studies.
Culturing of Microorganisms and Evaluation of
Antibacterial activity
e bacterial strains were maintained on agar slant at 4°C and activated
at 37°C for 24 h on nutrient agar (HiMedia, Mumbai, India) before any

Misra et al.: Pharmacognostic Standardization of Monochoria hastata (L.) Solms
536 Pharmacognosy Journal, Vol 10, Issue 3, May-Jun, 2018
=×
e
p
W
Extractive value(%) 100
W
Where, We = Weight of dried extract and Wp = Weight of plant powder
sample.
Qualitative Assessment of Phytochemical Constituents
Phytochemical compounds present in the 50% methanol extract were
assessed through the phytochemical screening tests following specic
protocols with some minor modication.36
Fluorescence Analysis
Fluorescence analysis of leaf powder was carried out by standard
protocol.37 In this analysis the plant sample was treated with various
basic and acidic solvents such as aqueous and alcoholic sodium hydroxide
(NaOH), hydrochloric acid (HCl), sulphuric acid (H2SO4), nitric
acid (HNO3), ferric chloride (FeCl3) and dibasic sodium bi phosphate
(Na2HPO4) and then observed in UV/ visible chamber under visible,
short UV-A wavelength (366 nm) and long UV-B wavelength (254 nm)
of light simultaneously.
Preparation of Ash and Estimation of Various Ash values
3gm of leaf powder was incinerated in a Silica crucible over the heater.
e charred mass was heated in mue furnace at 600-650ºC until the ash
became white and free from carbon. It was cooled and weighed on the
ash less lter paper. e water soluble ash, acid insoluble ash, sulphated
ash and total ash were determined following the standard protocol.38
Estimation of Moisture Content
e loss on drying was determined by estimating the moisture content
evaporated on drying at a temperature not exceeding 115ºC as per the
procedure put forwarded by e Indian Pharmacopoeia.39 e amount
of moisture was calculated using the following formula:
−
=×
Ww Wd
% moisture content (Wb) 100
Ww
Where, Ww = Wet weight of sample and Wd = Dry weight of sample.
Determination of Foreign Organic Matter
100g of leaf powder sample was weighed and spread on a tray in a thin
layer. e sample was inspected with the unaided eye and also with the
use of a 6x lens, and the foreign organic matter was separated manually.
Separated foreign matter was weighed and the percentage value was
determined as per Indian Pharmacopaea.40
CHNS and Heavy Metals Contents Analysis
Total carbon, hydrogen, nitrogen and sulphur contents were determined
using a CHNS analyser (Make: ermo Fisher Scientic Instruments;
Model: FLASH 2000 CHNS/O Analyzers; Country: USA), and lead and
arsenic contents were determined by ICP-MS according to the method
described by Mandal et al. (2017).41
Statistical Analysis
All the experiments except elements and heavy metals analysis have been
done in triplicate and the results accepted as Arithmetic Mean ± Standard
Error at 5% level.42
RESULTS
Antibacterial Evaluation
e dierent solvent extracts and puried fraction of M. hastata (L.)
Solms leaf exhibited antibacterial ecacy by showing bacterial zone
inhibition as shown in the Figure 2 (A-C).
Organoleptic Evaluation
e leaves showed green leathery appearance from both side without
any trichome. e leaf powder was green in colour, rough in texture,
mildly aromatic in odour and insipid in taste. e stem covered with the
remains of old leaf sheaths at the base was white, prostate to erect, so,
spongy and herbaceous in nature.
Macroscopic Evaluation
Morphological study indicated that leaves were simple, arrow shaped
or hastate with smooth margin, petiole was hollow, up to 3-4  long.
e lamina had parallel venation. Surface was smooth. e size of the
leaf varied from 45 to 60 cm in width and length, respectively. Root
was adventitious, laterally grown in tus from the tip of the rhizomatous
stem. Stem was short rhizomatous, simple covered with leaf sheath.
Inorescence was 6-9 cm long shortly stalked spike with 25-60 densely
arranged 13-16 mm long brilliant purple blue sub-umbellate raceme
owers (Figure 3 A-C).
Figure 2: Antibacterial evaluation of M. hastata (L.) Solms leaf.
(A) against S. mutans; (B) against S. aureus; (C) against E. coli. Here,
‘1’ indicates bacterial zone inhibition by 50% methanol extract,
‘2’ indicates bacterial zone inhibition by puried ethyl acetate fraction,
‘3’ indicates bacterial zone inhibition by absolute ethyl acetate extract
and ‘4’ indicates bacterial zone inhibition by antibiotic drug.
Figure 3: Macroscopy of M. hastata (L.) Solms. (A) Whole plant; (B) Leaf
with inorescence; (C) Inorescence.

Misra et al.: Pharmacognostic Standardization of Monochoria hastata (L.) Solms
Pharmacognosy Journal, Vol 10, Issue 3, May-Jun, 2018 537
Figure 5: Microscopy of M. hastata (L.) Solms leaf petiole. (A) T.S. of
petiole showing cuticle, epidermis and other visceral parts; (B) T.S. of
petiole showing vascular bundles and air-chambers; (C) T.S. of petiole
showing central air canal with diaphragm; (D) Presence of raphides.
Figure 6: Powder microscopy of M. hastata (L.) Solms leaf. (A) Sclereidal
bre; (B) Pitted tracheid, pitted vessel and raphide; C. Needle shaped
calcium oxalate crystals; (D) Annular vessel.
Figure 4: Microscopy of M. hastata (L.) Solms leaf. (A) Transverse section
(T.S.) of lamina; (B) T.S. of lamina showing epidermis, chlorenchyma tissue
and vascular bundles; (C) T.S. of midrib.
Leaf and Powder Microscopy
T.S. of lamina showed that the upper and lower both the epidermis
contained stoma. Wide air spaces were present in the median part. Epidermal
cells were covered with a very thin cuticle. Collateral vascular bundle
showed wide meta-xylem and narrow proto-xylem elements enclosed in a
parenchymatous sheath. Vascular bundle of mid-rib showed wide circular
thin walled meta-xylem with circular mass of phloem (Figure 4 A-C).
T.S. of petiole showed thick cuticle and ground tissue including palisade
layer followed by 3 layers of air-chambers divided by thin uniseriate
partition laments. Collateral vascular bundles were located in the inter-
cepts of the partition laments. A central air canal was surrounded by
a thin membranous plate of circular cells, called diaphragm. Needle
shaped raphides with pointed ends were also observed (Figure 5 A-D).
Powder microscopy of M. hastata (L.) Solms leaves indicated the presence
of sclereidal bres, pitted tracheids, pitted vessels, calcium oxalate crystals
and annular vessels (Figure 6 A-D).
Determination of Extractive values
e extractive values calculated from the yields in dierent solvents are
shown in the Figure 7. e extractive values in dierent solvents indicated
that water and 50% methanol extract showed the highest extractive
values while n-hexane and diethyl ether extract showed the lowest extrac-
tive values compared to other solvents. Chloroform, dichloromethane
and ethyl acetate had more or less same extractive value which is lesser
than water and methanol but greater than n-hexane and diethyl ether.
Phytochemical, Proximate and Fluorescence analysis
Phytochemical constituents present in the 50% methanol extract were
phenols, avonoids, saponins, steroids, carbohydrates, glycosides and
alkaloids. e results of proximates and uorescence analysis are presented
in Table 2 and Table 3, respectively.

Misra et al.: Pharmacognostic Standardization of Monochoria hastata (L.) Solms
538 Pharmacognosy Journal, Vol 10, Issue 3, May-Jun, 2018
DISCUSSION
e crude extracts and the puried fraction have potential antibacterial
ecacy against both Gram-positive and Gram-negative bacteria. e
amphistomatic and hydromorphic characters, features like presence of
cuticle, collateral vascular bundles, sclereidal bres, pitted tracheids,
pitted vessels, calcium oxalate crystals, annular vessels are the major
identifying characters of M. hastata (L.) Solms as the crude drug
source.43 Sclereidal bres conrm the presence of woody material in
the crude drug component, and also indicates its purity. Low moisture
content suggests better stability of the powder sample against microbial
degradation.44 e unique uorescence characteristic features are helpful
to assess the purity level of the powder drug sample. Ash content analysis
showed very signicantly lesser amount of acid insoluble ash than that
of water soluble ash which ensures the presence of negligible amount
of contaminants like silica, carbonates etc. Carbon, hydrogen and nitrogen
ratio (C: H: N: S) was 33: 6: 5: 1. M. hastata (L.) Solms contains a signi-
cant amount of nitrogen and sulphur which was not detected in another
aquatic medicinal herb, Eclipta prostrata, yet it contains amine and
nitrile compounds.45 is is an indication to denitely have nitrogenous
compounds in M. hastata (L.) Solms. e powder sample was completely
free from any organic impurities. Lead content in the leaf powder sample
was within the permissible limit while the arsenic content was higher
than the upper permissible limit as recommended by the Department
of Ayurveda, Yoga and Naturopathy, Unani, Siddha and Homeopathy
(AYUSH) for medicinal plants of India.46 It may be due to bioaccumu-
lation of arsenic absorbed by this aquatic plant from the water body
where it grew. In Malda, arsenic (As) concentration in ground water is
higher (0.05 – 1.434 mg/Lt) than the permissible limit of BSI (0.05mg/Lt).
Arsenic aected development blocks are English Bazar, Kaliachak-I,
Kaliachak-II, Kaliachak-III, Manikchak, Ratua-I and Ratua-II.47 Hence,
this particular aquatic herb may be used in bioremediation and restoration
of arsenic polluted wetlands and from the herbal use point of view the
plant sample must be checked before use as the arsenic content is high.
CONCLUSION
In can be concluded that M. hastata (L.) Solms leaf has potential antibac-
terial activity. e heavy metal contents analysis suggests that appropriate
safety measures should be taken before use of this herb as medicine. is
study will be helpful for authentication of M. hastata (L.) Solms as the
Elements and Heavy Metals Analysis
Carbon, hydrogen, nitrogen and sulphur (CHNS) contents of the leaf
powder sample were 27.85 %, 5.25 %, 4.31 % and 0.85 %, respectively.
Lead and arsenic contents in the powder sample are shown in the Figure
8.
Figure 7: Extractive values of M. hastata (L.) Solms leaf powder in dierent
solvents.
Figure 8: Lead (Pb) and Arsenic (As) contents of M. hastata (L.) Solms
leaf powder.
Table 2: Proximate analysis of M. hastata (L.) Solms powder sample.
Proximates Contents
Tot al as h 28%
Acid insoluble ash 2%
Water soluble ash 15%
Sulphated ash 8%
Moisture content 8.65%
Foreign organic matter 0%
Table 3: Fluorescence analysis of powder of M. hastata (L.) Solms leaf.
Experiments Visible/Day
light
UV Light
254 nm 365nm
Powder only Green Brown Black
Powder + 1N NaOH
(aqueous) Ye l l ow Green Black
Powder + 1N NaOH
(alcohol) Ye l l o w Brown Black
Powder + 1 N HCl Ye l l o w Brown Black
Powder + 50% H2SO4Light Brown Dark brown Black
Powder + Nitric acid Orange Brown Black
Powder + Ferric chloride Ye l l o w Green Black
Powder + Na2 HPO4Brown Dark
brown Black

Misra et al.: Pharmacognostic Standardization of Monochoria hastata (L.) Solms
Pharmacognosy Journal, Vol 10, Issue 3, May-Jun, 2018 539
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crude drug source. It will also help further researchers to maintain the
standards of this plant for their research projects. is pharmacognostic
standardization study will be used in pharmaceutical preparations if
necessary to recognize of M. hastata (L.) Solms from its adulterants. It
ensures the identity, quality and purity of the crude drug for the human
welfare. is study may be a diagnostic tool for using M. hastata (L.)
Solms as a phytoremediator of arsenic.
ACKNOWLEDGEMENTS
We are thankful to Edward Food Research and Analysis Centre (EFRAC)
for conducting the analysis of elements and heavy metals contents of the
plant sample.
CONFLICT OF INTEREST STATEMENT
We declare that we have no conict of interest.
ABBREVIATIONS USED
AYUSH:Ayurveda, Yoga and Naturopathy, Unani, Siddha and
Homeopathy; CHNS/O: Carbon, hydrogen, nitrogen, sulphur / oxygen;
FAO : Food and Agriculture Organization; ICP-MS: Inductively
coupled plasma mass spectrometry; ISM: Indian Systems of Medicine;
LOX: Anti-lipooxygenase; MHA: Mueller Hinton agar; MTCC:
Microbial Type Culture Collection and Gene Bank; WHO: World
Health Organization.
SUMMARY
M. hastata (L.) Solms is an evergreen perennial aquatic herb growing
in wide geographical range. ough the plant has been used by certain
ethnic communities in Asia, but the characterization of active principles
have not been done so far. e present research article focuses on the
evaluation of pharmacognostic identity and quality parameters to be
used as herbal drug and thus validating its ethno-medicinal use to cure
boils, gastritis, hepatopathy and as laxative.
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Misra et al.: Pharmacognostic Standardization of Monochoria hastata (L.) Solms
540 Pharmacognosy Journal, Vol 10, Issue 3, May-Jun, 2018
GRAPHICAL ABSTRACT SUMMARY
• M. hastata (L.) Solms is an evergreen perennial aquatic herb growing in wide
geographical range. Though the plant has been used by certain ethnic com-
munities in Asia, but the characterization of active principles have not been
done so far. The present research article focuses on the evaluation of pharma-
cognostic identity and quality parameters to be used as herbal drug and thus
validating its ethno-medicinal use to cure boils, gastritis, hepatopathy and as
laxative.
Debabrata Misra is a Ph. D. Research Scholar at the Plant and
Microbial Physiology and Biochemistry Laboratory, Department
of Botany, University of Gour Banga, Malda in West Bengal. He
He is currently working on purication and characterization of
antibacterial compounds from folklore aquatic plant for their use
in gastrointestinal micro ora.
Manab Mandal is a Ph. D. Research Scholar at the Plant and
Microbial Physiology and Biochemistry Laboratory, Department
of Botany, University of Gour Banga, Malda. He is currently en-
gaged in the research on high altitude plant resource as antibac-
terial agents.
Narendra Nath Ghosh is a Ph. D. Research Scholar at the De-
partment of Chemistry, University of Gour Banga, Malda. His re-
search focuses on the theoretical and computational chemistry
of uorescent dyes.
Vivekananda Mandal, Professor of Department of Botany, Uni-
versity of Gour Banga, Malda has more than ten years experi-
ence in the area of probiotics and antibiotic research. Dr Mandal
had published more than 50 research papers in national and in-
ternational journals and conferences.
ABOUT AUTHORS
Cite this article: Misra D, Mandal M, Ghosh NN, Mandal V. Pharmacognostic Standardization of an Ethnomedicinal Aquatic Herb,
Monochoria hastata (L.) Solms for its Antibacterial Potentiality. Pharmacog J. 2018;10(3):533-40.
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review. Int. J Biodiver.Conserv. 2012;4(3):101-12.
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terization of bringaraj and guduchi herbs by ICP-MS analysis, optical absorption
and EPR spectroscopic methods. Spectrochim. Act. Part A: Mol. Biomol. Spect.
2008;71(1):31-38.
46. Anonymous. 2007. http://www.qcin.org/documents/Certication_Scheme_AYUSH/.
47. Panigrahi S. Groundwater Arsenic Contamination: A Study of Malda District of
West Bengal, India. Imp. J Interdiscip. Res. 2016;2(8):742-7.