Pharmacognostic studies on Rumex Vesicarius

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Asian Journal of Plant Science and Research, 2011, 1 (1):102-115
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Pharmacognostic Studies on Rumex vesicarius
K. N. V. Rao
*
, Sunitha Ch, David Banjii, Sandhya S and Saikumar P
Department of Pharmacognosy, Nalanda College of Pharmacy, Nalgonda, Andhra Pradesh, India
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ABSTRACT
Rumex vesicarius is a branched succulent herb which belongs to the family Polygonaceae and is
distributed in India. The whole plant is medicinally important and cures several diseases.
Therefore in this context the detailed pharmacognostic study of various parts like
leaf,stem,petiole and root has been carried out with the aim to establish its Pharmacognostical
standards. The parameters selected were microscopical studies, proximate analysis, fluorescence
analysis and preliminary phytochemical screening including thin layer chromatographic
analysis.The microscopical studies of leaf,stem and root revealed the presence of sessile
glandular trichomes, spiral and scalariform xylem vessels.In physico-chemical evaluation the
ash values and extractive values were studied. Fluorescence analysis performed showed the wide
range of fluorescence colours for the crude powder as well as the extracts. The powder of
R.vesicarius was successively extracted with hexane,benzene,chloroform,ethylacetate,ethanol
and water for the identification of the best solvent. Preliminary phytochemical screening was
carried out for all the extracts and maximum chemical constituents was observed in the
ethanolic extract. The inorganic elementary analysis performed revealed the presence of
sodium,chloride and iron.The TLC studies performed produced fluorescent spots for the solvent
systems selected for flavonoids,steroids and anthraquinones .
Keywords: Scalariform , Fluorescence studies, adulterants , flavonoids, sessile glandular
trichomes.
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INTRODUCTION
Standardisation of a drug means confirmation of its identity and determination of its quality and
purity and detection of nature of adulterant by various parameters like morphological,
microscopical,physical observations. Standardisation and evaluation of herbal drugs mainly done
by two method they are organoleptic and botanical characters. In the organoleptic studies like
colour,odour,tast and texture where as in botanical microscopic and macroscopic characters are

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observed[1,2].Rumex the ancient Latin name for the docks or sorrels. vesicarius, vesica, a
bladder; from the inflated pods following the flowers on these herbs.It is a pale ,green
,dichotomously branched,succulent herb.Leaves are fleshy, sour, alternate, elliptic-ovate,broadly
ovate,entire,acute (or) obtuse,cordate at base,long petiole.Flowers are white,monoecious. Fruits
are nutlets,seeds are erect and trigonous [3] .Several C-glycosides,Flavonoids and
Anthraquinones are known to be constituents of this plant. The folklore claims that the plant is a
potent diuretic, astringent, carminative, stomachic and tonic[3,4,5]. Anti-bacterial and Anti-
oxidant activities of Rumex vesicarius was performed.[6,7] So far there is no pharmacognostic
report so this paper is aimed to report the pharmacognostic and phytochemical studies.
MATERIALS AND METHODS
Rumex vesicarius Plants were collected from local market in Nalgonda. It was identified and
authenticated by Prof. A. Lakshma Reddy, Retired Professor, Dept. of Botany, Nagarjuna Govt.
College (Autonomous) Nalgonda. The plant herbarium was prepared and deposited in the Dept.
of Pharmacognosy for further reference. The plant was identified as Rumex vesicarius Linn.
(Polygonaceae) under the voucher no: NCOPNLG/ph’cog/2010-2011/034.
Instruments Used
Micro senior precision rotary microtome (latest Spencer 820 type), Sisco muffle furnace
(3003137),Rotary vacuum evaporator,Hot air oven.
Chemicals and Reagents
All the chemicals and reagents like chloral hydrate, phloroglucinol, Hexane, Benzene,
Chloroform, Ethylacetate, Ethanol, used were of analytical grade.
Anatomical studies:
Transverse Section of Leaf, Stem and Root:[8-11]
Microtome Section was done for leaf,stem and root to obtain a thin section. The sections were
stained with phluroglucinol and hydrochloric acid in the ratio 1:1. Photo micrographs of different
magnifications were taken to study the anatomical features.
Powder Microscopy:[8,9]
Shade dried Leaf, Stem and root was powdered with the help of an electric grinder till a fine
powder was obtained. This fine powder was subjected to powder microscopy studies, as per
standard procedures mentioned.
Measurement of Cell Structure and Content:[8-10]
The length and width of phloem fibres,Length and Width of Trichomes, Width of Xylem
Vessels, Diameter of Calcium oxalate Crystals and diameter of the starch grains were measured
using stage micrometer and the eyepiece micrometer by standard methods.
Determination of Physico Chemical Parameters:[8,12,13]
Total ash, acid insoluble ash, water soluble ash, crude fiber content, moisture content, alcohol
soluble extractive value, water soluble extractive value, chloroform soluble extractive value and
petroleum ether soluble extractive values of Aerial parts of Rumex vesicarius were determined as
per standard procedures

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Leaf Constants:[8,17]
The stomatal number, stomatal index, vein islet number and vein termination number were
determined as per standard references.
Determination of Fluorescence Analysis:[12-15]
Powdered Leaf, Stem and root was subjected to analysis under ultra violet light after treatment
with various chemical and organic reagents.
Extraction
The collected aerial parts of the plant were washed and dried under the shade. Around 25 g of
the coarsely powdered aerial parts of the plant was packed in a soxhlet apparatus and
exhaustively extracted with the solvents of increasing polarity. The extract so obtained was
concentrated under vacuum using rotary vacuum evaporator and dried in dessicator until use.
Preliminary Phtyochemical Screening[8,13,15-19]
The extract so obtained were subjected to various chemical tests as per the procedure mentioned
in the standard reference books to determine the nature of chemical constituents present the in
the plant .
Thin Layer Chromatography Analysis[20,21]
Thin layer chromatography (TLC) is a chromatography technique used to separate mixtures of
chemical compounds. It is the most basic method of confirming the presence of a phytochemical
compound. The conditions maintained for performing the TLC analysis of the root are as
mentioned below:
Adsorbent: Precoated silical gel plates
Solvent systems:
STEROIDS: Benzene:Chloroform (3:7).
ANTHRAQUINONES: Ethyl acetate:Methanol:Water ( 100:13.5:10)
FLAVONOIDS: Benzene:Pyridine:Formic acid ( 72:18:10 )
Toulene:Dioxan:Glacial acetic acid ( 90:25:4 )
Chloroform:Methanol ( 9.3:0.6)
Detection by U.V at long wavelength
RESULTS AND DISCUSSION
ANATOMY OF THE LEAF: The leaf has dorsi-ventral orientation.
• Upper epidermis:Upper epidermis was single layer of thin walled closely arranged cells and
covered externally by a layer of cuticle. The upper epidermis was not a continous layer but slight
projection is seen in the centre .
• Lower epidermis:Lower epidermis was single layer of parenchymatous cells which is
interrupted by the stomata.The type of stomata is Anisocytic type of stomata.
• Mesophyll:The region between the upper and lower epidermis is constituted by
mesophyll.The mesophyll was formed of chlorenchymatous cells and is the seat of
photosynthesis.Mesophyll was differentiated in to upper palisade parenchyma and lower spongy
parenchyma.Palisade parenchyma consists of two or three layers of compactly arranged cells in
perpendicular to the surface.Spongy parenchyma consists of loosely arranged cells having air
filled spaces in between. The chloroplast are most abundant in Palisade cells than spongy
parenchyma.

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• Vascular bundles:Vascular bundles ranges from 2 to 4 in number.The pholem lies towards
the lower epidermis and the xylem lies towards the upper epidermis.
ANATOMY OF THE STEM:
EPIDERMIS
HYPODERMIS
XYLEM
PHLOEM
ENDODERMIS
CALCIUM OXALATE
CRYSTAL
CORTEX
Fig 2:- T.S of Rumex vesicarius Stem

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o Epidermis:Epidermis was outer most layerof compactly arranged cells. The cells are
elongated and rectangular and covered externally by cuticle. And the unicelluar trichomes was
seen on the epidermis region.
o Hypodermis:The hypodermis was inner to the epidermis and is formed of a few layers of
dead,thick walled sclerenchymatous cells.
o Cortex:The entire region inner to the hypodermis is called Cortex.It is formed of thin-walled
parenchymatous cells and having the large intercellular spaces in between them.The cortex
consists of a calcim oxalate crystals
o Vascular bundles:They are arranged as collateral and conjoint.Pholem towards the
epidermis and xylem towards the centre of ground tissue.And Pholem and Xylem are
differentiated by the endodermis and the cambium.
ANATOMY OF PETIOLE:
 Epidermis:It was the outer layer of the petiole and consists of oval or nearly circular cells of
a single layer which are thickened and slightly cutinized upon their exterior surface, and
presenting a fringed apperance.
 Collenchyma:They are arranged in several layers or thick angled cells underlyng the
epidermis.
 Parenchyma:Under the collenchymas there are a few layered parenchymatous cells.They
had a clear intercellular spaces.
 Vascular bundles:They are arranged in a collateral manner. They are in the form of v-shape.
Pholem and xylem is in the single bundle.
EPIDERMIS
COLLENCHYMA
PARENCHYMA
PHOLEM
XYLEM
Fig 3:- T.S of Rumex vesicarius Petiole

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ANATOMY OF THE ROOT:
 Cork:It was the outer most region of the root and is one cell thick in nature.The cells are
thin walled on the surface.
 Cortex:The region inner to Cork was formed of several layers of loosely arranged thin
walled parenchymatous cells.
 Pholem: Below the Cortex pholem is present .pholem consists of sieve tubes and pholem
fibres.
 Xylem:It is present in the middle region and xylem consists of Xylem vessels ,Xylem
parenchyma and xylem fibres.They are radially arranged.and the medullary rays are also present
in the xylem..Xylem vessels are in different radii.Xylem bundles are present alternately on
different radii number of each bundle ranges from 2 to 6 and the condition is called tetrarch.and
pith is usually absent.The diameter of the vessels in the xylem ranges from 113.6-28.4µ.
POWDER MICRSCOPY OF LEAF:
CORK
CORTEX
PHLOEM
MEDULLARY RAYS
XYLEM
Fig 5:-SPIRAL XYLEM VESSEL Fig 6:-UNICELLUAR TRICHOME
Fig 4:- T.S of the Rumex vesicarius Root

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Fig 7:-ANISOCYTIC STOMATA WITH
EPIDERMAL CELLS Fig 8:-SESSILE GLANDULAR TRICHOME
Fig 9:-CALCIUM OXALATE CRYSTAL Fig 10:-PARENCHYMA
Fig 11:-STRACH GRAIN

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POWDER MICROSCOPY OF STEM:
Fig 12:-NON-LIGNIFIED PHLOEM FIBRE Fig 13:-UNICELLULAR TRICHOME
Fig 14:-PARENCHYMA Fig 15:-BUNDLE OF PHLOEM FIBRES
Fig 16:
-
CALCIUM OXALATE CRYSTAL
Fig 17:-XYLEM VESSEL

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POWDER ANALYSIS FOR ROOT:
MEASUREMENTS:
This helps in identification of adulteration. The results obtained are tabulated in Table-1,2,3.
TABLE 1:-QUANTITATIVE MICROSCOPY OF LEAF
PARAMETERS LENGTH µm WIDTH µm
Trichome 37.5-217.35-462.5 12.5-32.5-62.5
Calcium oxalate crystal - 25-41.25-62.5
Xylem vessel - 25-38.125-75
Starch grain - 12.5-36.25-62.5
TABLE 2:-QUANTITATIVE MICROSCOPY OF STEM
PARAMETERS LENGTH µm WIDTH µm
Trichome 125-318.12-1587.5 12.5-28.12-50
Fibre 125-406.8-850 12.5-30-50
Xylem vessel - 25-52.12-150
Calcium oxalate crystal - 12.5-51.25-100
TABLE 3:-QUANTITATIVE MICROSCOPY OF ROOT
Determination of physico chemical properties:
The physico chemical properties help to estimate the amount of impurities like soil and particle
present in the drug. It also helps to assess the calculi salts present in the drug sample. Ash values
were used to detect the presence of any siliceous contamination and presence of any water
soluble salts .Alcohol and water soluble extractive values indicate the presence of the
adulterants, faulty processing and poor quality of the drug. While petroleum ether soluble
extractive value indicates percent of lipid content present in the crude drug.. Crude fibre content
is useful technique for differentiation of similar drugsThe results obtained for the proximate
analysis are tabulated in Table-4.
PARAMETER WIDTH µm
Xylem vessel 25-47.5-100
Fig 18:-SCALARIFORM XYLEM VESSEL Fig 19:-CORK CELL

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TABLE 4:-PHYSICO-CHEMICAL ANALYSIS OF AERIAL PARTS
PARAMETERS VALUES IN % W/W
Total ash 19.03
Acid insoluble ash 0.044
Water soluble ash 6.83
Sulphated ash 14.6
Water soluble Extractive 120
Ethanol soluble Extractive 51.40
Chloroform soluble Extractive 4.80
Pet-ether soluble Extractive 10.4
Moisture content 6.6
Crude Fibre Content 31.3
Determination of leaf constants
The stomatal number, stomatal index, vein islet &termination nos. obtained are tabulated in
Table-5
TABLE 5:-LEAF CONSTANTS OF RUMEX VESICARIUS
PARAMETERS UPPER SURFACE LOWER SURFACE
Stomatal number 12/sq.mm 15/sq.mm
Stomatal index 24 27.7
Vein Islet number 9
Vein Termination number 8
TABLE 6:- FLUROSCENCE ANALYSIS FOR LEAF POWDER
DRUG+REAGENT DAY LIGHT SHORT U.V
245nm LONG U.V
365nm
Drug+HNO
3
Yellowish Brown Light Green Dark Green
Drug+H
2
SO
4
Brownish Yellow Dark Green Fluroscent Green
Drug+NH
3
Light Green Fluroscent Green Dark Green
Drug+HCL Pale Green Fluroscent Green Black
Drug+Methanol Light Yellow Light Green Blackish Green
Drug+Fecl
3
Yellow Fluroscent Green Dark Green
Drug+5%NaOH Yellow Fluroscent Green Green
Drug+5%KOH Yellow Fluroscent Green Dark Green
Drug+1%KOH Yellowish Green Fluroscent Green Black
Drug+50%H
2
SO
4
Pale Green Pale Green Dark Green
Drug+50%HNO
3
Brown Fluroscent Green Dark Green
Drug+Methanolic NaOH Yellowish Brown Brown Yellow
Fig 20:-ANISOCYTIC STOMATA Fig 21:-VEIN ISLET

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Determination of fluorescence analysis
Fluorescence analysis is a tool to determine the kind of chemical nature of the drug. The
fluorescence obtained in short wavelength, long wave length and day light in table 6,7,8.
TABLE 7:-FLUORESCENCE ANALYSIS FOR STEM POWDER
DRUG+REAGENT
DAY LIGHT
SHORT U.V
245nm LONG U.V
365nm
Drug+HNO
3
Yellowish Brown Fluorescent Green Black
Drug+H
2
SO
4
Brownish Green Light Green Dark Green
Drug+NH
3
Yellow Light Green Dark Green
Drug+HCL Pale Yellow Pale Green Black
Drug+Methanol Pale Brown Light Green Yellow
Drug+Fecl
3
Yellowish Green Fluorescent Green Black
Drug+5%NaOH Pale Brown Light Green Dark Green
Drug+5%KOH Yellow Fluorescent Green Dark Green
Drug+1%KOH Pale Yellow Pale Green Dark Green
Drug+50%H
2
SO
4
Pale Brown Pale Green Black
Drug+50%HNO
3
Pale Yellow Pale Green Black
Drug+Methanolic NaOH Pale Yellow Greenish Yellow Yellow
TABLE 8:-FLUORESCENCE ANALYSIS FOR ROOT POWDER
DRUG+REAGENT
DAY LIGHT SHORT U.V
245nm LONG U.V
365nm
Drug+HNO
3
Yellow Light Green Black
Drug+H
2
SO
4
Brown Light Green Dark Green
Drug+NH
3
Reddish Brown PaleGreen Light Green
Drug+HCL Pale Yellow Pale Green Black
Drug+Methanol Pale Brown Pale Greenish Yellow Brownish Black
Drug+Fecl
3
Yellow Fluorescent Green Dark Green
Drug+5%NaOH Brown Pale Brown Brown
Drug+5%KOH Brown PaleGreen Dark Green
Drug+1%KOH Brown Pale Yellow Dark Green
Drug+50%H
2
SO
4
Brownish Black Pale Green Light Brown
Drug+50%HNO
3
Pale Brown Pale Green Brownish Black
Drug+Methanolic NaOH Dark Brown Brownish Black Black
TABLE-9:-FLUORESCENCE ANALYSIS OF EXTRACTS
EXTRACTS LONG WAVELENGTH SHORT WAVELENGTH DAYLIGHT
Hexane extract Black Green Reddish Brown
Benzene extract Black Green Reddish Brown
Chloroform extract Black Fluorescent Green Brown
Ethyl acetate extract Black Fluorescent Green Reddish Brown
Ethanol extract Black Dark Brown Brown
Aqueous extract Brown Greenish Brown Reddish Brown
EXTRACTION
• Extraction was carried out by the soxhaltion based on the increasing order of polarity as
follows:

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TABLE 10:-NATURE AND %YIELD OF DIFFERENT EXTRACTS
EXTRACTS COLOUR CONSISTENCY %W/W
Hexane Greenish Black Oily 3.46
Benzene Greenish Yellow Gummy 3.04
Chloroform Greenish Yellow Oily 2.092
Ethylacetate Greenish Black Waxy 1.824
Ethanol Reddish Green Oily 12.604
Water Brown Amophorous 26.092
Preliminary phytochemical screening:
Chemical test helps in the confirmation of the chemical nature of the active principles present in
the plant extract. The results of the chemical tests are tabulated in Table-10.
TABLE 11:-PRELIMINARY PHYTOCHEMICAL SCREENING OF RUMEX VESICARIUS
TEST HEXANE BENZENE CHLOROFOM ETHYLACETAE ETHANOL WATER
Carbohydrates - - + - - +
Proteins - - - - - -
Aminoacids - - - - - -
Fats and Oils + + + - - -
Steroids + + - - - -
Glycosides + + + + + -
Anthraquinone + + + + + +
Flavonoids - - - + + -
Alkaloids - - - - - -
Saponins - - - - + +
Tannins and
PhenolsCompounds - - - - - -
TABLE 12:-TEST FOR INORGANIC ELEMENTS OF THE POWDER
Test OBSERVATION
Test For Calcium -
Test For Magnesium -
Test For Sodium +
Test For Potassium -
Test For Iron +
Test For Sulphate +
Test For Chloride +
Test For Phosphate +
Test For Carbonate -
Test For Nitrate -
TABLE 13:- TLC ANALYSIS OF RUMEX VESICARIUS
PHYTOCONSTITUENTS SOLVENT SYSTEMS RF VALUES COLOUR OF SPOTS
Steriods Benzene:chloroform (3:7) 0.4
0.3
0.1
Orange
Orange
Orange
Anthraquinones Ethylacetate:methanol:water(100:13:5:10) 0.66
0.55 Brown
Brown
Benzene:Pyridine:Formic acid
( 72:18:10 )
0.4
0.75
0.65
0.82
0.95
Yellow fluorescence
Orange
Orange
Yellow fluorescence
Yellow fluorescence

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Flavonoids
Toulene:Dioxan:Glacial acetic acid
( 90:25:4 )
chloroform:Methanol ( 9.3:0.6)
0.93
0.81
0.84
0.25
Orange
Orange
Yellow fluorescence
Yellow fluorescence
CONCLUSION
Rumex vesicarius is widely cultivated in India for its culinary purposes. The microscopical
studies, physico chemical parameters,fluorescence analysis and chemical tests performed will
guide in the proper identification of the plant species from other species of Rumex as well as help
in authentication of the purity of the plant.By above all these parameters we can build up a
suitable plant profile.
Acknowledgement
The authors are grateful to the Principal and management of Nalanda College of Pharmacy in
providing all the support and help to carry out the work in the library and laboratory of the
college.
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