Pharmacognostical Evaluation and Phytochemical Screening of Neolamarckia Cadamba
Rubi Verma*, Fatma Chaudhary and Jayendra Kumar
School of Pharmacy, Monad University, India
Submission: March 23, 2019; Published: April 22, 2019
*Corresponding author: Rubi Verma, School of Pharmacy, Monad University NH-24, Hapur Road, Delhi, India
How to cite this article: Rubi Verma, Fatma Chaudhary, Jayendra K. Pharmacognostical Evaluation and Phytochemical Screening of Neolamarckia Cadamba. J Complement Med Alt Healthcare. 2019; 9(2): 555760. DOI:10.19080/JCMAH.2019.09.555760
The Pharmacognostical studies of Neolamarckia Cadamba (Roxb.) Leaf the purpose of identification and differentiation from related species. The macroscopic and microscopic features of the Leafs were studied, including the use of powder microscopy with the aid of suitable tools and reagents. Physicochemical parameters such as ash value, extractive value and weight loss on drying were also determined. The Leafs powder was successively extracted with different solvents followed by preliminary phytochemical screening of the extracts. Preliminary phytochemical screening of different extracts revealed the presence of alkaloids, carbohydrate, protein, gum, steroid, tri-terpenoids, saponin, flavonoids and tannin in the Leafs. The scientific parameter is necessary to identify the exact plant material and to find its quality and purity. These studies indicated the possible information for correct identification and standardization of this plant material.
Neolamarckia Cadamba Miq; syn. Anthocephalus Kadamba (Family: Rubiaceae) commonly known as ‘Kadamba’ in Ayurveda1 is a large deciduous tree between 37.5-45meter height. The stem of younger trees appears greyish-green with smooth bark. As it gets older, the bark gets rough and grey with longitudinally fissured. Leafs glossy, dark green, opposite, simple pulvinus base sub sessile to petiolate, broadly ovate to elliptical-oblong, entire, apex marinate and venation pinnate. The flowers that appear from August to October are orange to yellow. Inflorescence in clusters, terminal globose heads, subsessile and fragrant. Fruitlets numerous with upper parts containing 4 hollow or solid structures. Seed trigonal or irregularly shaped. The trees found in the greater part of India in moist localities in West Bengal, Bihar, Andhra Pradesh, Karnataka, Kerala and peninsular India [1-2].
The plant finds its application in several traditional and folklore systems of medicine around the globe. The tribal of India use the Leafs paste orally against dyspepsia and locally applied in mouth ulcer in children. Leafs are nutritious, astringent and their decoction is reported to be used for gargling in aphthae or stomatitis . Dried powdered Leafs used as anthelmintic and the tribal people of India used hot water extract of the Leafs as an astringent, stomatitis and for washing wounds inthroat [4-8]. The flowers are used as vegetable and as gurgle to remove the foul smell from mouth. The fruit is cooling and said to destroy the phlegm and impurity of blood when eaten. The ripened fruit are aromatic, acidic with astringents property. Lod has taken ripened fruits as carminative/masticate. Fruit juice is given during fever and gastric disturbance. Similarly stem barks reported to possess astringent, febrifuge, antiseptic and acts as diuretics. Juice of the bark given orally against cough, fever and in inflammation of eyes. Dried stem bark also used as folk medicine (ethno medicines) in the treatment of various skin diseases, anemia, uterine complaints and for improvement of semen quality. Lod has apply stem bark paste on swelling of legs and juice to cure eye inflammation.
The fresh juice of the bark is applied to the heads of infant when the fontanels sunken. Mundas prescribe the bark paste duly suspended in water in reducing blood sugar in the patients with diabetes mellitus . Several pharmacological and biological tests have been reported on this plant are evident from literatures. Alcoholic extracts of dried Leafs possess analgesic, anti-inflammatory antimicrobial, wound healing, antioxidant and anti- alarial antibacterial and antifungal, activities. However, only a few phytochemicals have been reported on this plant in the literature types of sapogenins such as cadambagenicacid, quinovic acid and ß-sitosterol was isolated from the bark.
Few alkaloids are also reported from the bark and Leafs like
cadambine, 3α-dihydrocadambine and isohydrocadambine .
In the light of all the above and keeping the medicinal overview
of N. Cadamba, the present investigation was being carried
out to study some Pharmacognostical features of the Leafs as
a whole including its intact and powdered form available in
the literature . The studies were carried out in accordance
with WHO General Guidelines for Herbal Drug Standardization
methodologies. The findings from this study would be useful
as standards for the species as well as a source of reference for
further scientific investigation of the species .
The Leafs of the Neolamarckia Cadamba was collected from
Monad University Hapur, Uttar Pradesh. For authentification, I
made a herbarium in which plant part are attached. Then it was
authentified from the taxonomist of NISCAIR, New Delhi. After
authentication, fresh Leafs were collected in bulk, washed with
potable water to remove adhering dirt followed by rinsing with
distilled water, and then shade dried and powdered .
The following macroscopic characters for the fresh and
dried Leafs were noted: surfaces, size and shape, fracture,
texture, color, odour and taste. Leafs are slightly aromatic with
unpleasant taste. Anatomical characteristics of the Leafs Fresh
Leafs pieces were subjected to dehydration procedure from
aqueous and alcohol .
The shade dried powdered Leafs screened through sieve no.
40 was used for the powdered drug analysis. The specimens were
separately treated with glycerin, N/20 iodine solution, 10 % w/v
alcoholic ferric chloride (for detection of phenolic compounds),
phloroglucinol-hydrochloric acid (1:1) for detecting lignin and
ruthenium red solution (for detection of mucilage). After staining,
the samples through temporary micro slide preparation taking
the mount ant glycerin and were observed under a compound
The dried and powdered Leafs (50g) was successively extracted
with petroleum ether (60-80°C), chloroform, ethanol
and water by reflux for 24h by Soxhlet apparatus. Following extraction,
the liquid extracts were concentrated under reduced
pressure using rotary evaporator to yield dry residues. The extracts
were subjected to preliminary phytochemical screening
using standard procedures to determine the nature of phytoconstituents
content . The result of the preliminary phytochemical
screening of different extracts (Table 1) showed presence ofalkaloids
(in chloroform and methanol extracts), carbohydrates,
proteins, gum (in aqueous extract), steroids (in petroleum ether,
chloroform and ethanol extracts), triterpenoid (in petroleum
ether and chloroform extracts), saponin (in chloroform, ethanol
and aqueous extracts) and flavonoids and tannin (in ethanol and
aqueous extracts) .
The physicochemical parameters including ash values (total
ash, acid insoluble ash, water soluble ash and sulphated ash),
extractive values (ethanol, ether and water soluble) and loss on
drying were performed according to the standard treatises .
Leafs coriaceous, entire margin, elliptical-oblong or ovate,
pulvinus base, with acute or shortly acuminate. It is often used
in the form of powder (nygrodhadi kvatha churna) which is a
Isolated fragments of uniseriate conical hairs either whole
or broken are found. Few, whole unicellular conical hairs, pieces
of epidermis of lower surface with wavy anti clinical walls and
stomata; few pieces of isolated stomata and prismatic crystal of
calcium oxalate are found in the microscopy.
The microscopic study of Neolamarckia Cadamba leaf
showed the presence of simple elongated, unicellular trichomes,
rubiaceous types of stomata on the lower side of the leaf, starch
grains, crystals of calcium oxalate, wedge-shaped vascular
bundles, and phloem in the form of ring and oil globules. The leaves of Neolamarckia Cadamba having methyl salicylates
aroma when crushed by hands.
TLC was performed to develop phytochemical finger
printing. It was performed using 2x10cm TLC plates coated by
silica gel G. 10μl Volume of each extract was applied on plates
with the help of capillary a thin layer (0.25mm). In addition, a
binder like gypsum is mixed into the stationary phase to make
it stick better to the slide. TLC plate on the side with the white
surface draw a thin line with pencil. The thin end of the spotter
is placed in the dilute solution; the solution will rise up in the
capillary (capillary forces). Touch the plate briefly at the start
line. Allow the solvent to evaporate and spot at the same place
again. This way you will get a concentrated and small spot. A
TLC plate can be developed in a beaker. Place a small amount
of solvent (mobile phase) in the container. The solvent (eluent)
travels up the matrix by capillarity, moving the components of
the samples at various rates because of their different degrees
of interaction with the matrix (stationary phase) and solubility
in the developing solvent. Non-polar solvents will force nonpolar
compounds to the top of the plate, because the compounds
dissolve well and do not interact with the polar stationary phase.
Allow the solvent to travel up the plate until~1 cm from the top.
Take the plate out and mark the solvent.The components, visible
as separated spots, are identified by comparing the distances
they have traveled with those of the known reference materials.
Measure the distance of the start line to the solvent front. Then
measure the distance of center of the spot to the start line. Divide
the distance the solvent moved by the distance the individual
spot moved. The resulting ratio is called Rf – value.
The Rf (retardation factor) depends on the following
1. Solvent system
2. Absorbent (grain size, water content, thickness)
3. Amount of material spotted
The chromatograms were developed at room temperature
in a 10x10cm twin trough chamber using solvent systems
Toluene: Ethyl acetate in a ratio of 6:4 for the ethanolic extract of
Neolamarckia Cadamba. After the development chromatograms
of saponin were derivatized with 20% Antimony trichloride in
chloroform in a ratio of 20:100ml followed by heating at 110°c in
preheated oven for 10min. These chromatograms were scanned
and evaluated under wave lengths of 254nm & 366nm using a
camag TLC to get graphical representation of finger prints. From
the TLC finger printing of the ethanolic extract drugs, presence
of the saponins, alkaloids, glycosides, steroids, flavonoids as the
principle chemical compounds were identified.
A plant may be considered as a biosynthetic laboratory,
not only for the chemical compounds such as carbohydrates,
proteins and lipids that are utilized as food by man, but also for a
multitude of compounds like alkaloids, glycosides , steroids and
sterols, saponins, flavonoids, phenolic compounds and volatile
oils that exert a physiological effect. The compounds that are
responsible for therapeutic effects are usually the secondary
metabolites. A systematic study of a crude drug embraces through
consideration of both primary and secondary metabolites
derived as a result of plant metabolism in addition to its macro
and microscopic studies. N. Cadamba is often confused with other
species due to their relative similarities. The species has been
taxonomically described as distinct species by earlier workers.
The Leafs finds its application in several other traditional and
folklore systems of medicine around the globe that have been
previously described and surprisingly no pharmacopoeia
standards are available for them in the literature. Owing to its
importance in applications, the present study was designed and
conducted. From the present study, it can be concluded that the
macroscopic and microscopic findings together will help future
investigators in proper identification of the plant.Further, the
powder microscopy, preliminary phytochemical screening and
physicochemical parameters would aid in standardization of the
plant material. The wide spectrum of biological activity of this
plant is due to presence of several phytoconstituents that needs
to be studied further [14-21].
In the present study, leafs of Neolamarckia Cadamba Roxb.
Pharmacognostical Evaluation for the identification of various
Phytoconstituents and rest of extracts were utilized for pharmacological
screening. The various extracts after the Pharmacoganostics
Evaluation have shown the presence of following
active principles. Distilled water extract: Steroids, Glycosides,
Alkaloids, Tannins, Phenolic compounds, Flavonoids. From the
ongoing studies, it can be concluded that the above macroscopic
and microscopic studies together may be used as a tool for identification
of Neolamarckia Cadamba with its Pharmacognostical
characteristics, discriminating it from its other species diversity.
The authors are thankful to NISCAIR Herbarium for plant
identification (New Delhi) and grateful to the Dr. Jayendra Kumar
Principal of School of pharmacy, Monad University. Hapur U.P, for
providing necessary facilities to carry out this research.