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COMPARISON OF THE LEAF DRUG RUTA GRAVEOLENS AND ITS SUBSTITUTE

RUTA CHALEPENSIS

Article in International Journal of Research in Ayurveda and Pharmacy · December 2018

DOI: 10.7897/2277-4343.096180

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 Rubeena Mattummal et al / Int. J. Res. Ayurveda Pharm. 9 (6), 2018

Research Article
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COMPARISON OF THE LEAF DRUG RUTA GRAVEOLENS AND ITS SUBSTITUTE RUTA CHALEPENSIS
Rubeena Mattummal 1*, Divya Kallingilkalathil Gopi 1, Sunil Kumar Koppala Narayana 2
1
Research Assistant, Department of Pharmacognosy, Siddha Central Research Institute, Central Council for Research in
Siddha, Ministry of AYUSH, Govt. of India, Arumbakkam, Chennai 600106, India
2
Research Officer & HOD, Department of Pharmacognosy, Siddha Central Research Institute, Central Council for
Research in Siddha, Ministry of AYUSH, Govt. of India, Arumbakkam, Chennai 600106, India

Received on: 25/08/18 Accepted on: 25/10/18

*Corresponding author
E-mail: [email protected]

DOI: 10.7897/2277-4343.096180

ABSTRACT

Ruta graveolens is known as common rue, belongs to family Rutaceae. Rue oil is obtained from steam distillation of fresh plant material and used as
anthelminthic, antispasmodic, antiepileptic, rubefacient and emmenagogue. It’s a potential drug used in all the traditional systems of medicine. Aruvatha
mattirai and Aruvathac curanam are the Siddha formulations prepared from dried leaves of R. graveolens. The leaves of R. chalepensis commonly
known as fringed rue, is reported to be used as a substitute owing to similar morphology in dried form. So this current study is to compare and contrast
the authentic drug, R. graveolens from its substitute. Methods: Dried leaves of both species were collected from medicinal garden of Siddha Medical
College, Arumbakkam. The macroscopic and microscopic details including powder were studied following the standard pharmacopoeial procedures.
Results: Macroscopically leaves are different colored with dissimilar shape, in microscopic study R. graveolens showed toothed lamina with glandular
trichomes while R. chalepensis showed lamina without trichomes. Quantitative microscopy of both species was carried out and the epidermal number,
stomatal number, stomatal index, vein islet number, vein islet ratio, palisade ratio was recorded. Powder microscopy of R. graveolens leaves showed
the presence of pitted and spiral vessels while R. chalepensis showed annular and spiral vessels. Conclusion: The study provides the characteristics
features to distinguish R. graveolens from R. chalapensis.

Keywords: Adulteration, Powder microscopy, quantitative microscopy, Substitution

INTRODUCTION the presence of alkaloids, flavonoids, coumarins, tannins, volatile

oil, sterols and triterpenes8. The pharmacological activities like
Ruta graveolens L. is indigenous of Southeastern Europe but is antioxidant, anticancer and anti-inflammatory effects are reported
widely naturalized in Southern Europe and cultivated worldwide. from this species9.
It is a perennial shrub with smooth erect stems; leaves are green
colored compound leaves with lobed obovate leaflets, flowers are All Ruta species are associated with phyto photodermatitis10 and
found in terminal umbels with four spoon-shaped yellow shining plants should not be touched with bare hands, especially on sunny
non- fringed petals; fruit is a capsule containing many seeds1. R. days. In 14 species of genus Ruta; R. graveolens and R.
graveolens used in traditional medicines for the relief of pain, eye chalepensis are available in India and also cultivated in gardens.
problems, rheumatism and dermatitis for a long time. Medicinal The plants are explored very less for its medicinal properties.
properties like anti-inflammatory, analgesic, antibacterial, Even though the taxonomical characters to recognize the Indian
antidiabetic2 and antifertility3 effects of R. graveolens extracts species are very apparent in case of flower morphology,
have been reported in various studies. The phytochemicals being references to it are mystified in the traditional literature11. So the
discovered and isolated from the extracts are rutin, quercetin, anatomical study of the aerial parts of the plant can be a solution
rutacridone, gravacridondiol and the essential oil contains to mitigate this drawback for the proper identification and
monoterpenes like limonene, α-pinene and 1,8-cineole4. In authentication of the plants since both are used in traditional
Siddha leaves of Ruta graveolens are used in the preparation of systems of medicines, the adulteration or substitution of drugs can
Aruvatha mattirai and Aruvathac curanam5. The plant is not prevent to some extent.
mentioned in the classical Ayurveda books but described for
cough, cold and dysmenorrheal in late nighantus of 20th century. MATERIALS AND METHODS

Ruta chalepensis is indigenous to Mediterranean region and Botanically identified and authenticated leaves of Ruta
Canary islands6 is found growing on rocky places, dry banks and graveolens (Voucher No: SCRIP010) and Ruta chalepensis
thickets, usually found on limestones7 and is acclimatized world (Voucher No: SCRIP011) were procured from the medicinal
over. It is a perennial herb growing up to 80 centimeters tall. The garden of Siddha Medical College, Arumbakkam. The
leaves are compound; the inflorescence is a cluster of flowers, macroscopy of the samples were documented by Nikon
each with four or five bright yellow petals with rolled, fringed COOLPIX5400 digital camera. Sectioning of the fresh samples
edges. The fruit is a textured capsule which is divided into pointed were done to reveal the anatomy of the parts and the rest were
lobes. The plant is abortifacient, anthelmintic, emmenagogue and dried, powdered, passed through mesh no.60, and preserved in an
ophthalimic7. R. chalepensis is a rich source of important air-tight covers for powder microscopy. Transverse sections of
secondary metabolites such as furanocoumarins and alkaloids. the leaves were hand cut using a 7’o clock platinum blade stained
Phytochemical screening of the aerial parts of the plant showed with safranin and photographed using Nikon ECLIPSE E200

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trinocular microscope attached with Nikon COOLPIX5400 a group of phloem cells, and xylem facing towards pith. The pith
digital camera under bright field light. Quantitative studies were is formed of parenchyma cells (Fig. 2).
carried out according to standard procedure. Magnifications were
indicated by the scale-bars. A pinch of the powdered leaves were In R. chalepensis, the petiole appears circular in TS with single
mounted in glycerine on a clean microscopic slide. Slides were layered epidermis followed by a layer of hypodermis. The outer
observed under Nikon ECLIPSE E200 trinocular microscope and cortex is large and formed of 8 to 10 layers of chlorenchyma cells
diagnostic characters were identified, photographed and with randomly distributed volatile oil cells. The middle cortex is
documented12. made up of few layers of collenchyma cells containing rosette
crystals. A ring of 4 to 5 collateral vascular bundles are seen in
RESULTS AND DISCUSSIONS the inner cortical region similar to that of R. graveolens.
Parenchyma cells in the center forms the pith (Fig. 3).
Macroscopy
The leaves of R. graveolens are bluish green colored when fresh Lamina
and turn to yellowish brown when dried; leaves alternate, entire
and fern like compound with deeply lobed obovate leaflets, gland The TS of leaf of R. graveolens showed both upper and lower
dotted with dark spots; unipinnate to tripinnate; leaflets are linear epidermis with cuticle followed by mesophyll cells. The
to oval or oblong; 7.5 to 20 cm long and 2-3cm broad; leaves epidermis bears glandular trichomes on both surfaces giving a
become fragile on drying and mostly found as powdery in the toothed appearance to the epidermal layer. The mesophyll cells
bulk sample. The odor is strongly aromatic and tastes slightly consist of two layers of palisade cells and loosely arranged
bitter. spongy cells. Intervening the palisade layer some volatile oil cells
is present. In between palisade and spongy layers a good number
In R. chalepensis the leaves are, grey- green colored, 10 to 12 cm of rosette crystals of calcium oxalates are also found. The
long and 5 to 8 cm broad, compound and each divided into several vasculature is formed of xylem and phloem elements (Fig. 4).
segments which are subdivided into smaller leaflets; blong-
elliptical to linear shaped leaflets with prominent dark spots and In R. chalepensis, the outer thick walled epidermis is single
oil glands. The leaves have a strong and unpleasant smell tasting layered without having any trichomes. The mesophyll
bitter (Fig. 1). differentiated into upper double layered palisade and lower
spongy parenchyma cells which are organized loosely. In the
Microscopy midrib region a layer of few cells of palisade also found on the
lower epidermal region just below the vascular bundle. Volatile
Petiole oil cells can be seen on the palisade layer. Abundance of rosette
TS of petiole of R. graveolens is circular in outline with outer crystals of calcium oxalate is distributed in the lamina. The
epidermis followed by single layered hypodermis. The outer vascular bundle is formed of normal elements (Fig. 5).
cortex consists of 3 to 4 layers of chlorenchyma cells with few
volatile oil cells distributed randomly. The middle cortex is Quantitative microscopy
formed of 4 to 5 layers of collenchymas cells some of which
contains prismatic crystals and rosette crystals. The inner cortex The leaves of both species were subjected to quantitative
is made up of few parenchyma cells and a ring of 5 conjoint, microscopy. It revealed the presence of peltate glandular
collateral vascular bundles at the center. A discontinuous trichomes on both the surfaces of R. graveolens leaf and
pericycle layer surrounds the vascular ring which is followed by abundance of rosette crystals on R. chalepensis leaf (Fig. 6 & 7).
Table 1: Quantitative microscopy of Ruta graveolense and R. chalepensis

R. graveolens R. chalepensis
Parameters Upper (/mm2) Lower (/mm2) Upper (/mm2) Lower (/mm2)
Epidermal number 88 – 128 80 - 176 56 - 164 80 – 100
Trichome number 8 – 10 10 - 12 - -
Stomatal number 40 – 54 48 - 60 40 - 52 52 – 76
Stomatal index 29 – 31 25 - 38 24 - 41 40 – 43
Palisade ratio 3.25 - 5 6 - 6.75
Vein islets number 12 - 16 5–6
Vein termination number 29 - 39 15 – 20

Table 2: Comparison between R. graveolens and R. chalepensis

Characteristic features Ruta graveolens L. Ruta chalepensis L.

Macroscopy
Leaves
1. Colour Blue green Grey-green
2. Size 7.5 to 20 cm long, 2-3cm broad 10 to 12 cm long, 5 to 8 cm broad
3. Shape Obovate- linear to oval Oblong- elliptical to linear leaflets
Flowers Non-fringed petals Fringed petals
Fruits Capsule with blunted tips Capsule with pointed tips
Microscopy
Leaves
1. Lamina Toothed surface Smooth surface
2. Trichomes Glandular Absent
3. Rosette crystals Scares Abundant
Petiole
1. Cortex Narrow cortex with 2 to 3 layers of chlorenchyma Broad cortex with many layers of chlorenchyma
2. Prismatic crystals Present Absent

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Powder Microscopy
1. Colour Brownish yellow Yellowish brown
2. Odour Strongly aromatic Unpleasant
3. Taste Slightly bitter Bitter

Powder Microscopy photographs illustrated in the Fig 4.4 and 4.5 gives the greater
evidence of occurrence of trichomes on both the leaf surfaces of
The leaf powder of R. graveolens was brownish yellow with R. graveolens giving it a toothed appearance. The stomatal
characteristic aroma and tastes slightly bitter and showed diversity characteristics (type, shape, size and orientation) in the
characters like epidermis of petiole in surface view, leaf foliar epidermis have a great value in plants systematic and
epidermis in surface view, cortical parenchyma of petiole, spongy taxonomic studies17. The presence of anomocytic stomata in R.
parenchyma, pith parenchyma with vasculature, vessels with graveolens also reported in earlier records18,19 but they reported
annular and pitted thickening, fibers with wide lumen and xylem the occurrence only on the lower surface. In this study we could
parenchyma cells, vessel fragments and oil cavity cells (Fig. 8). observe obviously the amphistomatic distribution of stomata in
both species which similar to the findings of Dhale et al.,
The leaf powder of R. chalepensis was yellow brown with 201019.According to Metcafe and Chalk20 palisade ratio is a
unpleasant smell and bitter taste. It showed the characters like leaf reliable taxonomic character, which is constant for a taxon and it
epidermal cells in surface view, fragment of mesophyll cells, will not vary with environment. The palisade ratio we recorded is
spongy parenchyma, pith parenchyma with vasculature, vessels nearly same as findings of Dhale et al., 201019.The presence of
with annular and spiral thickenings, thick walled fibers with wide broad outer cortex with chlorenchyma layers in the petiole can
lumen, rosette crystals, starch grains and oil globules (Fig. 9). distinguish R.chalepensisfrom R. graveolenswhere only 2 to 3
layers are observed. The abundant of rosette crystals of calcium
The two species of Ruta growing in India are very difficult to oxalates in leaves is also a significant character in R. chalepensis
identify and differentiate since both are morphologically very while comparing with that of scares amount in R. graveolens. But
similar and having almost same anatomical features with R. graveolenspossess prismatic crystals in petiole whereas which
negligible differences. These were used interchangeably in is absent in R. chalepensis.
ancient times13 and now a day’s R. chalepensis is become an
adulterant for R. graveolens. The studies available in the literature CONCLUSION
is also confusing and contradicting each other and need a very
clear description and characterization of the two for the The pharmacognostic characters described to distinguish the Ruta
authentication of plant species. The comparison of macro and species will be a supportive tool to compare the plants. Since most
microscopic characters are being supportive for the better of the pharmacognostic publications pertaining to these plants are
discrimination between the two plants. The blue-green contradicting each other further DNA finger printing and
appearance of leaves in R. graveolens distinctly differentiates R. molecular studies are necessary to accomplish the identity and
chalepensis with grey green leaves and fringed petals of R. authenticity of the species.
chalepensis from non-fringed flowers of R. graveolens.
ACKNOWLEDGEMENT
Presence of essential-oil cavities mentioned as the characteristic
feature of Rutaceae14 was present in random distribution in both. The authors extend their heartfelt thanks to Director General,
Even though these plants are gland dotted and scented, they show Central Council for Research in Siddha, and Assistant Director,
a distinct difference in the odor and taste. This study gives the key Incharge, Siddha Central Research Institute, Chennai for the
distinguishing character of presence of glandular trichomes in R. support.
graveolens leaf which supports the works of Jayaweera15 while R.
Kannan et al., (2012)16 strongly disagrees the findings that they
couldn’t observed the trichomes even in both species. But the

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11. Al-Said MS, Tariq M, Al-Yahya MA, Rafatullah S, Ginnawi
OT, Ageel AM. Studies on Ruta chalepensis, an ancient Rubeena Mattummal et al. Comparison of the leaf drug Ruta graveolens
and its substitute Ruta chalepensis. Int. J. Res. Ayurveda Pharm.
medicinal herb still used in traditional medicine. J
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Source of support: Nil, Conflict of interest: None Declared
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