IJPBA_2059_23_20230717_V1.pdf

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RESEARCH ARTICLE
Pharmacognosy and Phytochemistry of Leaves of Tephrosia uniflora
H. C. Sahana, E. Akila, V. B. Narayanaswamy
Department of Pharmacognosy, RR College of Pharmacy, Bengaluru, Karnataka, India
Received: 28 February 2023; Revised: 20 March 2023; Accepted: 15 April 2023
ABSTRACT
The need for and significance of pharmacognostic analysis of leaf extracts of Tephrosia uniflora are
discussed in the current research. The various extracts like petroleum ether, chloroform, alcohol, and
aqueous leaf extracts of T. uniflora were prepared and studied for their organoleptic characteristics,
macroscopic, powder microscopic, physicochemical analysis, phytochemical analysis, and fluorescence
analysis of T. uniflora. The physicochemical evaluation shows that loss on drying was found to be
0.14% w/w, total ash value was 6.94% w/w, acid insoluble ash value was 3.16%w/w, and the extractive
value of petroleum ether extract was 3.2% w/w. Chloroform extracts were found to be 3.9% w/alcohol
extracts was found to be 4% w/aqueous extracts were found to be 4.8% w/Phytochemical analysis shows
the presence of carbohydrates, phenols, flavonoids, alkaloids, and tannins. These are just a few of the
parameters that need to be evaluated for a pharmacognostic study of T. uniflora.
Keywords: Fluorescence analysis, pharmacognostic study, physicochemical analysis, phytochemical
study, Tephrosia uniflora
INTRODUCTION
Traditional medicine, which includes herbal
medicines, has recently been defined by the WHO
as including therapeutic practices that were utilized
for hundreds or even thousands of years before
modern medicine’s rise and diffusion and are still
in use today. Traditional medicine is a synthesis
of the healing techniques used by generations
of indigenous doctors. The only conventional
medicines that can be considered “herbal drugs”
are those that primarily use medicinal plant
extracts. The use of herbal medicine is growing in
popularity because of the toxicity and side effects
of allopathic medicines. This led to a sudden
rise in the number of companies that produce
herbal medicines. Herbal treatments have been
the principal treatment method in traditional
medical systems since the dawn of civilization.
*Corresponding Author:
H. C. Sahana,
E-mail: sahanachandru29@gmail.com
The practices are still in use today because of their
biological benefits, importance to the preservation
of human health, and inclusion in many cultural
beliefs.[1,2]
Tephrosia uniflora is a semi-erect perennial
with silky stem hairs that can grow up to 1 m
tall. T. uniflora is an herb that grows on rocky
terrain and is sub-fruticose and abundantly
branching. slender and angular branches from the
base, pubescent growth. Most axillary or geminate
flowers are single. A few pink, pea-shaped flowers
appear in the leaf axils. Up to 4 mm long flower
stalks are common. Calyx has up to 6 mm-long
fangs and a tube that is covered with velvet-like
hair.About 1 cm long and externally hairy in velvet.
Compound leaves have leaf stalks that are 3–8 mm
long and an axis that is about 2 cm long. Leaflets
range in size from 5 to 9, are up to 5.5 cm long and
13 mm broad, have an inverted-lance or elliptic
form, are pointy or blunt, and are hairless or velvet-
hairy above and appressed hairy below. Stipules
can be up to 9 mm long. Pods measure between 3.8
and 4.8 cm in length, 4–4.5 mm in width, and 7–8
Available Online at www.ijpba.info
International Journal of Pharmaceutical Biological Archives 2023; 14(2):1-5
ISSN 2582 – 6050

Sahana, et al.: Pharmacognosy and phytochemistry of leaves of Tephrosia uniflora
IJPBA/Apr-Jun-2023/Vol 14/Issue 2 2
seeds in length. The Arabian Peninsula, Pakistan,
and NW India, particularly Rajasthan, are the
locations where rock Tephrosia is found. February
through March and August through November are
when flowers bloom.[3-7]
Tephrosia plants come in a wide diversity, and
many of them have had their chemical makeup
and pharmacological properties investigated.
Flavonoids, rotenoids, and sterols are among
T. uniflora’s key chemical classes. Flavonoids are
the substances that have been isolated and identified
the most frequently, which should be emphasized.
T. uniflora has been studied for its components,
which include elongatin (1), 12a-hydroxyrotenone
(5), sitosterol, and stigmasterol.[8,9]
In herbal medicine, T. uniflora is commonly found
and used to treat a wide range of ailments, such
as stomachaches, diarrhea, asthma, inflammation,
and respiratory issues. They’ve also been applied to
the treatment of snake bites. Prenylated flavonoids
and isoflavonoids produced by the genus have
been shown to have cytotoxic, antiplasmodial,
anticancer, antibacterial, and anti-inflammatory
properties. Some flavonoids, particularly
isoflavonoids, prevent bacteria from producing
DNA, metabolizing their food, or forming cell
membranes.[10]
By using step-by-step pharmacognostic research,
standardization can be accomplished. These
research projects aid in the identification and
authentication of the plant’s T. uniflora. To ensure
the reproducible quality of herbal medicine,
which will contribute to its safety and efficacy,
correct identification and quality assurance of
the raw materials are prerequisites that cannot be
ignored. Its morphological, powder microscopy,
phytochemical screening, and physico-chemical
properties are pharmacognostic approaches utilized
in the standardization of plant material.[11]
MATERIALS AND METHODS
Collection, Identification and Authentication of
Plant Material
The plant’s leaves, of T. uniflora, were collected.
After drying in the shade, it turns into a coarse
powder. The plant material that had been obtained
was acknowledged and confirmed by scientist
(Dr.) K Madhavachetty, M.Sc., M. Ed., M.Phil.,
PhD, PGDPD, assistant professor, Department of
Botany, Tirupathi, India.
Morphological and Microscopical Features
T. uniflora was examined under a microscope and
with the naked eye for macroscopical evaluation,
noting the leaf’s color, size, scent, and other
diagnostic characteristics. There were differences
in the leaves’ macroscopic characteristics.
Observing the type of leaf, form, arrangement,
apex, margin, venation, base, texture, etc. was
part of evaluating the leaves. Powder microscope,
T. uniflora’s full plant was coarsely ground up and
examined under a microscope. A chloral hydrate
reagent was used to macerate the powder. After
being macerated, the powder was colored using
phloroglucinol and HCl reagents. Glycerine was
used to mount small amounts of the dyed powders
on a slide. Photographs were obtained under a
photomicroscope of the various cellular features
and inclusions.[12]
Physicochemical Parameters
T. uniflora leaves have undergone physicochemical
analysis in accordance with the WHO and
pharmacopoeias’ recommendations. Some of these
requirements are total ash, acid-insoluble ash,
water-soluble ash, extractive values that are water-
soluble, and extractive values that are alcohol-
soluble.[13-15]
Preparations of Extracts
Extract was made using the previously powdered
medication. Different extracts are made by
extracting plant material with increasing amounts
of polarity from Pet ether, Chloroform, Ethanol,
and Water. About 50 g of the air-dried powdered
plant material were consecutively extracted in a
Soxhlet apparatus with petroleum ether (40–60°),
chloroform, and ethanol. Water extraction was
carried out by maceration. The marc was air dried
below 50°C each time before extraction using

the following solvent: With the aid of Whatmann
filter paper, the extracts were purified, the solvent
evaporated at room temperature, and precise weight
measurements were taken. In order to calculate
the extractive value (%), air-dried medication was
used.
Preliminary Phytochemical Screening
To determine the presence or absence of
major primary and secondary metabolites like
carbohydrates, protein, alkaloids, steroids, phenol,
glycosides, terpenoids, and flavonoids, among
others, a preliminary qualitative phytochemical
screening of plant extract from T. uniflora leaves
was performed using standard methods.[16-18]
Fluorescent Analysis
T. uniflora leaf powders and extracts were treated
with various solvents and reagents to produce
distinct fluorescence characteristics, which were
then observed in the visible light, short UV
(254 nm), and long UV (366 nm) spectrums.[19]
RESULTS AND DISCUSSION
Morphology of T. uniflora Leaves
T. uniflora is an herb that grows on rocky terrain
and is sub-fruticose and abundantly branching.
slender and angular branches from the base,
pubescent growth. Most axillary or geminate
flowers are single. A few pink, pea-shaped flowers
appear in the leaf axils. Calyx has up to 6 mm-
long fangs and a tube that is covered with velvet-
like hair. About 1 cm long and externally hairy in
velvet. Compound leaves have leaf stalks that are
3–8 mm long and an axis that is about 2 cm long.
Leaflets range in size from 5 to 9, are up to 5.5 cm
long and 13 mm broad, have an inverted-lance or
elliptic form, are pointy or blunt, and are hairless
or velvet-hairy above and appressed hairy below.
Stipules can be up to 9 mm long. Pods measure
between 3.8 and 4.8 cm in length and 4–4.5 mm in
width [Figure 1].
Microscopic Characteristic of Powder
T. uniflora of Leaves
Inthisactivity,powderedplantmaterialisviewedunder
the microscope (Magnification ×45). All the lignified
cells stained pink color. Calcium oxalate crystals were
observed under the polarized light microscope. The
powder characters are listed in Figure 2.
Table 2: Yield of extracts obtained from successive
extraction of leaves of Tephrosia uniflora
Plant
name
Type of
extract
Appearance/State Yield
(% w/w)
Tephrosia
uniflora
leaves
Pet ether Yellowish green/Semisolid 3.2
Chloroform Greenish black/Semisolid 3.9
Ethanol Darkgreen, black/Semisolid 4
Water DarkBrown black/Semisolid 4.8
Table 1: Physicochemical constants of leaves of Tephrosia
uniflora
S. No. Parameters Percentage yield (% w/w)
1. Losson drying 0.14
2. Acidinsoluble ash 3.16
3. Sulphated ash 2.6
4. Watersoluble ash 5.3
5. Total ash 6.94
Table 3: Preliminary phytochemical screening of various
extracts of Tephrosia uniflora
Chemical
tests
Tephrosia uniflora leaf extracts
Pet ether Chloroform Ethanol Water
Proteins and
Aminoacids
‑ ‑ ‑ ‑
Carbohydrates ‑ ‑ + +
Steroids ‑ ‑ ‑ ‑
Phenols ‑ ‑ + +
Saponins ‑ ‑ ‑ ‑
Flavonoids ‑ ‑ + +
Alkaloids ‑ ‑ + +
Glycosides ‑ ‑ ‑ ‑
Tannins ‑ ‑ + +
+: Indicates presence and –: Indicates absence
Figure 1: Morphological characters of Tephrosia uniflora of
leaves
Whole plant leaves and flowers

Table 4: Fluorescence analysis of powder leaves of Tephrosia uniflora
S. No. Treatment Day light Short UV (254 nm) Long UV (366 nm)
1. Powder Green Green Green
2. Powder+Water Green Light green Light green
3. Powder+1NHCl Light green Light green Bluish white
4. Powder+1NH2
SO4
Pale green Light green Bluish green
5. Powder+1NHNO3
Pale yellowish green Light green Yellowish green
6. Powder+Aceticacid Dark green Dark green Red
7. Powder+1NNaOH Yellowish green Light green Dark green
8. Powder+1N Alc.NaOH Yellowish green Light green Dark green
9. Powder+1NKOH Green Light green Dark green
10. Powder+1N Alc.KOH Light green Light green Dark green
11. Powder+Ammonia Green Light green Dark green
12. Powder+Iodine Reddish brown Dark green Dark green
13. Powder+Fecl3
Brownish green Dark green Dark green
14. Powder+Ethanol Light green Light green Red
Table 5: Fluorescence analysis of various extracts of
Tephrosia uniflora
S.
No.
Extracts Daylight UV light
Short 254 nm Long 365 nm
1 Pet ether Green Yellowish green Reddish
2 Chloroform Greenish black Dark green Reddish
3 Ethanol Greenish black Greenish black Reddish
4 Water Brownish black Green Greenish
Determination of Physicochemical Constants
The physico-chemical constants of Tephrosia
unifora leaf parts were determined for loss of
drying, Ash value, and Extractive value as per
the method described in pharmacopoeias, and the
results are mentioned in Tables 1 and 2.
Preliminary Phytochemical Screening of
Extracts
Preliminary phytochemical investigations of
extracts revealed the presence of different
secondary metabolites. Ethanol and aqueous
extracts indicated the presence of flavonoids,
carbohydrates, alkaloids, phenols, and tannins,
respectively. The result is given below in
Table 3.
Fluorescence Analysis
The selected plant is made into a course powder,
treated with the required chemical reagents, and
observed under visible and ultraviolet rays; the
results are given in Tables 4 and 5.
Figure 2: Powder microscopy of leaves of Tephrosia
uniflora (a) Trichomes, (b) Calacium oxlate crystals,
(c) Xylem vessels, (d) Lamina with vascular tissue,
(e) Browinsh matter, (f) Epidermal cells alinged with
stomata, (g) Epidermal cells with stomata
b
a
c d
e f
g

CONCLUSION
T. uniflora plant leaves were evaluated for
pharmacognostic characterization, measurement
of physiochemical parameters, and phytochemical
screening of the crude extracts in the current study.
The chosen plants were examined macroscopically
and microscopically to confirm their authenticity
and purity. As part of the usual procedure,
physicochemical analyses such as ash value, acid-
insoluble ash value, and extractive value were
performed.
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