Collection, Extraction and Phytochemical Analysis of Indian Borage Leaves

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International Journal of Plant Pathology and Microbiology 2024; 4(2): 52-56

E-ISSN: 2789-3073
P-ISSN: 2789-3065
www.plantpathologyjournal.com Collection, extraction and phytochemical analysis of
IJPPM 2024; 4(2): 52-56
Received: 22-07-2024 Indian borage leaves (Plectranthus amboinicus)
Accepted: 26-08-2024

Mohammed Asif Killedar Mohammed Asif Killedar, Sowmya KL and Ramalingappa B


Research Scholar, Department
of Microbiology, Davangere
University, Shivagangotri, DOI: https://doi.org/10.22271/27893065.2024.v4.i2a.93
Davangere, Karnataka. India
Abstract
Sowmya KL This study focuses on the collection, extraction, and phytochemical analysis of the leaves of
Research Scholar, Department Plectranthus amboinicus, commonly known as Indian borage. The leaves were subjected to a
of Microbiology, Davangere systematic extraction process using methanol, followed by filtration and concentration to obtain the
University, Shivagangotri, extracts. Phytochemical screening revealed the presence of various bioactive compounds, including
Davangere, Karnataka, India alkaloids, glycosides, tannins, saponins, terpenoids, and flavonoids, while fixed oils and fatty acids
were found to be absent. The extracted compounds are noted for their potential pharmacological
Ramalingappa B
properties, such as antioxidant, antibacterial, anticancer, and antiplatelet activities, highlighting the
Senior Professor, Department
therapeutic potential of Plectranthus amboinicus in traditional medicine. The methodology involved
of Microbiology, Davangere
University, Shivagangotri, drying the plant material, grinding, and soaking in methanol, ensuring a comprehensive extraction of
Davangere, Karnataka. India phytochemicals. This research contributes to the understanding of the medicinal value of Indian borage
and supports its use in herbal remedies.

Keywords: Plectranthus amboinicus, ethyl acetate extraction, phytochemical analysis

1. Introduction
Indian Borage, also known as Plectranthus amboinicus or Coleus amboinicus, is a versatile
and aromatic herb native to tropical regions of Africa and Asia, particularly India (Ashaari et
al., 2021) [5]. It's commonly known by several names, including Cuban Oregano, Mexican
Mint, and Spanish Thyme. Indian Borage features thick, fleshy, and ovate leaves with a
distinctive aromatic scent. The plant is typically bushy and can grow up to 1-2 feet in height.
The leaves are green and have a somewhat wrinkled texture, and they often bear a slightly
fuzzy surface. The leaves have a strong, pungent aroma that is reminiscent of oregano and
thyme. Indian medicine has long used Plectranthus amboinicus (Lour) Spreng, popularly
known as Indian borage. It is sometimes referred to as country borage in English and is a
member of the Lamiaceae family (Kirtikar and Basu., 1999) [16]. Many medical conditions
can be treated with a decoction of the leaves, most notably respiratory conditions like
congestion, bronchitis, sour throats, and digestive disorders including diarrhea, colitis,
dysentery, and so on. The leaves of P. amboinicus have been shown to possess a wide range
of pharmacological characteristics, including urolithiasis, antiepileptic, antitumorogenic,
antimutagenic, radioprotective, neuropharmacological, and antibacterial capabilities (Khare
et al., 2011) [15]. Significant antioxidant efficacy, reducing power, nitric oxide radical
scavenging, superoxide anion radical scavenging, and ferrous ion chelating capacity are all
displayed by the leaves (Kumaran and Karunakaran., 2006) [17]. Gram-positive and Gram-
negative bacteria are inhibited by the essential oils, flavonoids, and terpenes found in them
(Gurge et al., 2009; Bhatt and Negi., 2012) [12, 9]. The leaves of Indian borage have also been
shown to exhibit antimutagenic, antitumorogenic, and antigenotoxic properties (Annapurani
and Priya., 1999) [4]. While a wealth of material exists on P. amboinicus leaves, little is
known about the plant's stem. Against this backdrop, the current study was conducted to
ascertain the phytochemical composition of P. amboinicus stem and to assess some
functional features of the plant using various in vitro models.

Correspondence 2. Materials and Methods


Mohammed Asif Killedar
Research Scholar, Department
2.1 Collection and Extraction of P. amboinicus
of Microbiology, Davangere The leaves of P. amboinicus were collected from the plants grown in the campus of
University, Shivagangotri, Shivagangothri, Davangere University, and Davangere. The plant material was recognized
Davangere, Karnataka. India by Dr. Ramalingappa B., Senior Professor., Department of Microbiology.

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International Journal of Plant Pathology and Microbiology https://www.plantpathologyjournal.com/

Leaves of P. amboinicus in requisite quantities were washed persistent froth is obtained. The persistence of froth for 10
and cut into ∼2 cm length pieces and ground by using mins signifies the presence of saponins.
mortar and pestle. The solvent (ethyl acetate, 30 ml) was
combined with the fresh leaves extract (10 ml), by using the 2.2.5 Test for Terpenoids and Phytosterols (Salkowski
separating funnel method, obtained the ethyl acetate extract test): To 0.5 ml of test solution, add equal quantity of
(Owolabi et al., 2018) [20]. The obtained greenish brown chloroform and then add sulphuric acid along the sides of
extract was filtered through Whatman No.1 filter paper, and the test tube avoiding shaking. The appearance of reddish
the light brown filtrate was used for chemical analysis. The brown color in the chloroform layer signifies presence of
test was carried out on the same day when the leaves were phytosterols.
collected.
2.2.6 Test for Flavonoids
2.2 Phytochemical Screening of P. amboinicus extract 2.2.6.1 Lead acetate test: To 1 ml of test solutions, add 1
The test sample was subjected to phytochemical analysis in ml of 10% lead acetate solution. The appearance of yellow
order to find out the presence of phytochemical constituents colored precipitate signifies the presence of flavanoids.
according to Praveen Bhatt et al., 2012 [9].
2.2.6.2 FeCl3 test: To 2 ml of test solutions, add ferric
2.2.1 Test for Alkaloids chloride solutions drop by drop. The formation of greenish
2.2.1.1 Mayer's test: To 2 ml of test solutions, few drops of black color indicates presence of flavonoids.
Mayer’s reagent (potassium mercuric iodide solution) was
added and observed for turbidity or precipitation. The 2.2.7 Test for Proteins (Biuret test): Add 2 ml of Biuret
appearance of cream colored precipitate signifies the reagent to 2 ml of test solutions. Shake well and warm it on
presence of alkaloid. water bath. Appearance of red or violet color indicates
presence of proteins.
2.2.2 Test for Cardiac Glycosides (Keller-Killiani test):
To 2 ml of test solutions, 1 ml of glacial acetic acid and 1-2 2.2.8 Test for fixed oils and fatty acid (Spot test): A drop
drops of ferric chloride solution was added. Then 0.5 ml of of Extract was added to the filter paper and observed. Oil
concentrated sulphuric acid was slowly added along the staining on the filter paper indicates the presence of fixed oil
sides of the test tube avoiding shaking of test tube. A and fats.
reddish brown ring at the interface of two liquids indicates
the presence of deoxysugar characteristic of cardenolides. 3. Results and Discussion
3.1 Collection of the sample
2.2.3 Test for Tannins and Phenolic compounds (Lead The leaves of Indian borage were collected from campus of
test): To 2 ml of test solutions, 1-3 drops of Ferric chloride Shivagangothri, Davangere. It was packed with polythene
was added and the mixture was observed for blue or green bag and brings to lab for experiment purpose. Then, the
color. leaves washed with tap water and it was disinfected
removed the water contains by the tissue paper and used for
2.2.4 Test for Saponins (Foam test): The test solutions are the experiment purpose as shown in the Fig.1 (Sindhu and
taken in a test tube and shaken vigorously till a stable Poonkothai., 2021) [22].

A B C
Fig 1: Collection of Indian borage leaves from campus of Shivagangothri, A- P. amboinicus; B –Collected P. amboinicus in lab condition;
C- P. amboinicus washed with distilled water

3.2 Extraction by Ethyl acetate method distinguishing the leaf pigments and oil (Abdulkadir and
The fresh leaf of Indian borage was crushed with the help of Ali., 2015) [1]. The leaf oil which also referred as ethyl
mortar and pestle to get the leaf juice. Brown colour leaf acetate extract, which will use for the investigations of
juice is subjected to separating funnel by taking 10 ml of phytochemical analysis as shown in the Fig.2.
juice and Ethyl acetate about 30 ml in the funnel, which will

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International Journal of Plant Pathology and Microbiology https://www.plantpathologyjournal.com/

A B C D E
Fig 2: Steps of Indian borage leaves extraction method. A- Fresh P. amboinicus leaves; B- Maceration of P. amboinicus using Morter and
Pistle; C- Extract of P. amboinicus ; D- P. amboinicus and ethyl acetate (1:3) in separating funnel; E-Ethyl acetate extract.

3.3 Phytochemical analysis of P. amboinicus compound was identified using specific reagents that
The analysis confirmed the presence of alkaloids, produced characteristic colour changes or reactions (Zahari
glycosides, tannins, saponins, terpenoids, flavonoids, and et al., 2020) [26]. Phytochemical analysis was conducted to
proteins through various chemical tests (Janakiraman et al., determine antioxidant compounds as shown in the Fig. 3
2023) [14]. Notably, fixed oils and fatty acids were absent as and the results were tabulated in Tab.1.
indicated by the lack of oil staining on filter paper. Each

A B C D

E F G H
Fig 3: Phytochemical analysis report of P. amboinicus A- Alkaloids; B- Glycoside; C- Tannis and Phenolic compounds; D- Saponin; E-
Terpenoids and Phytosterols; F- Flavonoids; G- Protein; H- oils and fatty acid

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International Journal of Plant Pathology and Microbiology https://www.plantpathologyjournal.com/

Table 1: Phytochemical analysis of Plectranthus amboinicus leaves.


Test solution Crude Extract
Phytochemical Test
Observation Inference
Alkaloids Mayer’s test Cream colour ppt +
Glycoside Keller Killiani test Brown ring +
Tannins and Phenolic compounds Lead test Green colour +
Saponins Foam test Foam persists for 10 mins +
Terpenoids and Phytosterols Salkowaski’s test Reddish- Brown ring +
Lead acetate test Yellow ppt +
Flavonoids
FeCl3 Greenish –black colour +
Proteins Biuret’s test Violet colour +
Fixed oil and fatty acids Spot test No staining -

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