Kapil et al., (2025).
Plants and Secondary Metabolites, Volume 6; ISBN: 978-81-984754-0-4
Chapter 10
Exploring the phytoconstituents of flowers
Eragrostis unioloides (Retz.) Nees ex Steud.,
through TLC profiling & phytochemical
screening
Manas Jyoti Kapil1, Bhagwati Prashad Sharma2, Nibedita Jena3* and
Sanjeet Kumar3
1
Department of Pharmaceutics, Royal School of Pharmacy, The Assam Royal Global
University, Guwahati, Assam, India
2
Department of Botany, Sidharth Government College Nadaun, Himachal Pradesh, India
3
Ambika Prasad Research Foundation, Odisha, India
Email-Id: [email protected]
ORCID: https://orcid.org/0009-0000-0238-0412
DOI: https://doi.org/10.5281/zenodo.15206120
Licensed under a Creative Commons Attribution 4.0 International License
ABSTRACT
Eragrostis unioloides, a grass species, has been traditionally used for various purposes. To explore its potential
medicinal properties, a phytochemical screening and thin-layer chromatography (TLC) analysis were conducted.
The results revealed the presence of tannins, saponins, reducing sugars, and alkaloids in different extracts. The TLC
analysis showed distinct Rf values for various compounds in different mobile phases. This study lays the
groundwork for future research into the medicinal properties of E. unioloides and its potential applications in
traditional medicine. The findings highlight the importance of screening grass species for bioactive compounds,
which could lead to the discovery of new medicines.
Keywords: Bioactive compounds, grass, therapeutic agents
INTRODUCTION
Grasses are one of the most diverse and widespread groups of plants, with many
species being used in traditional medicine for various purposes (Gebashe et al.,
2020). Despite their importance, grasses remain an underexplored source of
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bioactive compounds (Gono et al., 2022). Recent studies have highlighted the
potential of grasses as a source of novel medicines, with many species showing
promising pharmacological activities (Zeng et al., 2018; Mukarram et al., 2021;
Kiełtyka-Dadasiewicz et al., 2024). Eragrostis unioloides, a species of grass, has
been traditionally used in various cultures for its medicinal properties (Saxena and
Brahmam, 1996; Siddabathula and Prasanna, 2023; Panda and Panda, 2012).
However, there is limited scientific evidence to support its use, and further research
is needed to explore its potential as a source of bioactive compounds (Nayak et al.,
2024). This study aims to investigate the phytochemical composition of E.
unioloides flowers and to identify potential bioactive compounds that may be
responsible for its medicinal properties (Aldughaylibi et al., 2022). The search for
bioactive compounds in grasses is an important area of research, as it may lead to
the discovery of new medicines and provide insights into the pharmacological
activities of these compounds (Dincheva et al., 2023). This study will contribute to
our understanding of the phytochemical composition of E. unioloides and its
potential applications in traditional medicine (Sinha et al., 2023).
METHODOLOGY
The flowers of E. unioloides were collected from the Athagarh division of Cuttack
district in Odisha, India (Figure 3). The plant species was identified by the authors
(Figure 1-2). The Soxhlet extraction for phytochemical analysis was carried out and
nine secondary metabolites were detected using standard methods (Devi et al., 2023;
Marndi et al., 2024; Mety et al., 2024).
Test for tannin: 1 ml of flower extract was taken in a test tube, and 3-5 drops of
10% lead acetate solution were added to it. The gelatinous precipitate formation
confirmed the presence of tannin.
Test for saponin: About 1 ml of distilled water was added to 1 ml of the flower
extract and shaken well. The formation of persistent froth was observed, confirming
the presence of saponin.
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Figure 1: Flowers of E. unioloides from Odisha state, India
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Figure 2: Flowers of E. unioloides from Himachal Pradesh, India
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Figure 3: Collected flowers of E. unioloides for experimental work
Test for flavonoids: About 1 ml of the flower extract was taken. 2 ml of NaOH
solution and 3-5 drops of dilute HCl were added to it. The colour initially turned to
an intense yellow colour with NaOH solution and later became colourless, which
confirmed the presence of flavonoids.
Test for terpenoids: To 1 ml of the flower extract, 5-6 drops of chloroform were
added. Then, 4-6 drops of concentrated sulphuric acid were added to it. The reddish-
brown interface confirmed the presence of terpenoids.
Test for phenols: About 1 ml of the flower extract was taken. A few drops of 5%
ferric chloride solution were added. The dark bluish-black appearance confirmed
the presence of phenolic compounds.
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Test for reducing sugars: About 1 ml of the flower extract was taken. Two drops
of Fehling’s solution A, followed by Fehling’s solution B, were added and kept in
the water bath for a few minutes. The presence of red-orange precipitate confirmed
the presence of reducing sugars.
Test for steroids: To 1 ml of the flower extract, 1 ml of chloroform followed by 1
ml of concentrated sulphuric acid were added. The appearance of upper red and
lower yellow color with green fluorescence provides the presence of steroids.
Test for alkaloids: About 1 ml of the flower extract was taken. 3-4 drops of
Dragendroff’s reagent were added to it. The formation of a reddish-brown
precipitate confirmed the presence of alkaloids.
Test for carbonyl compounds: About 1 ml of flower extract was taken, and 3-4
drops of 2,4- Dinitrophenylhydrazine (DNPH) reagent was added. The formation of
yellow crystals confirmed the presence of carbonyl compounds.
TLC (Thin-layer chromatography) analysis
TLC plates were prepared on 6 cm glass slides using silica gel powder. Slides were
washed with clinical laboratory detergent and dried. Clean and dried slides were
wiped with ethyl acetate for removing surface adherents. 3 g of silica was taken in
20 ml of distilled water, and slurry was prepared by constant stirring and was finally
poured over the slides and slides were left undisturbed till the drying of the silica
layer. Then, the slides were activated at 50ᵒ C for 15-20 minutes before running the
TLC. Methanol: chloroform (1:1), ethyl acetate: chloroform (1:9), and ethyl acetate:
distilled water: acetic acid (8:1:1) were used as mobile phases. Then spots were
marked, the RF (Retention Factor) values were recorded and visualized under the
UV chamber (Kumar and Jena, 2014; Mishra and Bhatnagar, 2024).
RESULTS AND DISCUSSION
The phytochemical screening of E. unioloides flowers revealed the presence of
various bioactive compounds, including tannins, saponins, reducing sugars, and
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alkaloids (Figures 4-6, Table 1). The presence of these compounds suggests that the
plant may have potential medicinal properties. The TLC analysis showed distinct Rf
values for various compounds in different mobile phases, indicating the presence of
a range of bioactive compounds with different polarities. The results suggest that E.
unioloides may have potential applications in traditional medicine, particularly in
the treatment of diseases where these compounds have shown efficacy. The presence
of tannins and saponins in the plant suggests that it may have antimicrobial and anti-
inflammatory properties, while the presence of alkaloids may indicate potential
analgesic and anti-inflammatory activities. The reducing sugars detected in the plant
may also contribute to its medicinal properties.
TLC Results and Polarity of Mobile Phases: The TLC analysis of E. unioloides
flowers was conducted using different mobile phases with varying polarities (Figure
7, Table 2). The results are as follows:
1. Chloroform: Methanol (1:1): This mobile phase has a moderate polarity.
The Rf values of 0.28 and 0.46 for the acetone extract suggest that the
compounds present in the extract have moderate polarity, allowing them to
move up the plate but not too far. The Rf value of 0.93 for the hydro-
ethanolic extract suggests that the compounds present in this extract are
relatively non-polar.
2. Ethyl acetate: Chloroform (1:9): This mobile phase has a relatively low
polarity. The Rf values of 0.79 and 0.87 for the acetone extract suggest that
the compounds present in the extract are relatively non-polar, allowing them
to move up the plate. The Rf value of 0.91 for the hydro-ethanolic extract
suggests that the compounds present in this extract are also non-polar.
3. Ethyl acetate: Distilled water: Acetic acid (8:1:1): This mobile phase has
a relatively high polarity. The Rf values of 0.93 and 0.96 for the acetone
extract suggest that the compounds present in the extract are relatively polar,
allowing them to move up the plate. The Rf values of 0.75 and 0.83 for the
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hydro-ethanolic extract suggest that the compounds present in this extract
are also polar but to a lesser extent.
Overall, the results of the present study provide a foundation for further research
into the medicinal properties of E. unioloides and its potential applications. Further
studies are needed to isolate and characterize the bioactive compounds present in
the plant and to evaluate their pharmacological activities.
Table 1: Qualitative phytochemical screening of Eragrostis unioloides flowers using
different extracts
Bioactive compounds Extracts
n-Hexane Ethanol Aqueous
Tannin Not detected Detected Detected
Saponin Detected Not detected Detected
Flavonoids Not detected Not detected Not detected
Terpenoids Not detected Not detected Not detected
Phenols Not detected Not detected Not detected
Reducing sugar Not detected Detected Detected
Steroids Not detected Not detected Not detected
Alkaloids Not detected Detected Detected
Carbonyl compounds Not detected Not detected Not detected
Table 2: TLC analysis of Eragrostis unioloides flowers using different mobile
phases
Mobile phases Extracts Rf values of spots
S1 S2
Chloroform: Acetone 0.28 0.46
Methanol (1:1)
hydroethanolic 0.93 Nil
Acetone 0.79 0.87
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Ethyl acetate: Hydroethanolic 0.91 Nil
chloroform
(1:9)
Ethyl acetate: Acetone 0.93 0.96
Water: Acetic
acid (8:1:1) Hydroethanolic 0.75 0.83
Figure 4: Detection of secondary metabolites of E. unioloides flowers using n-hexane extract
Figure 5: Detection of secondary metabolites of E. unioloides flowers using ethanol extract
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Figure 6: Detection of secondary metabolites of E. unioloides flowers using aqueous extract
Figure 7: Spots on TLC plates under UV light of E. unioloides acetone extracts using
mobile phases; a) (Chloroform: methanol), b) (Ethyl acetate: chloroform) and c) (Ethyl
acetate: water: acetic acid)
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During the literature survey, it was noticed that there are fewer or no reports
available on the E. unioloides flower, but researchers have documented the
medicinal values and bioactive compounds of grass species. Sujatha et al., (2022)
reported the high cytotoxicity activity of ethyl acetate extracts of E. amabilis and E.
pilosa. Jabeen et al., (2023) reported the phytochemical and allelopathic potential
of invasive grass species.
CONCLUSION
The phytochemical screening and TLC analysis of Eragrostis unioloides flowers
revealed the presence of various bioactive compounds, including tannins, saponins,
reducing sugars, and alkaloids. These findings suggest that E. unioloides may have
potential medicinal properties, warranting further research into its pharmacological
activities and potential applications. The study highlights the importance of
screening grass species for bioactive compounds, which could lead to the discovery
of new medicines.
REFERENCES
Aldughaylibi FS, Raza MA, Naeem S, Rafi H, Alam MW, Souayeh B, Farhan M, Aamir M,
Zaidi N and Mir TA. (2022). Extraction of bioactive compounds for antioxidant,
antimicrobial, and antidiabetic applications. Molecules (Basel, Switzerland).
27(18): 5935. DOI: 10.3390/molecules27185935
Devi RS, Satapathy KB and Kumar S. (2023). Validation of tribal claims for formulation of
future drugs through evaluation of ethno-pharmacological values of Ludwigia
adscendens. Medicinal Plants. 15(4): 691-697.
Dincheva I, Badjakov I and Galunska B. (2023). New insights into the research of bioactive
compounds from plant origins with nutraceutical and pharmaceutical
potential. Plants (Basel, Switzerland). 12(2): 258. DOI: 10.3390/plants12020258
Gebashe F, Aremu AO, Gruz J, Finnie JF and Van Staden J. (2020). Phytochemical profiles
and antioxidant activity of grasses used in South African traditional
medicine. Plants (Basel, Switzerland). 9(3): 371. DOI: 10.3390/plants9030371
94
Kapil et al., (2025). Plants and Secondary Metabolites, Volume 6; ISBN: 978-81-984754-0-4
Gono CMP, Ahmadi P, Hertiani T, Septiana E, Putra MY and Chianese G. (2022). A
comprehensive update on the bioactive compounds from seagrasses. Marine
Drugs. 20(7): 406. DOI: 10.3390/md20070406
Jabeen S, Ali MF, ud Din AM, Javed T, Mohammed NS, Chaudhari SK, Javed MA, Ali B,
Zhang L and Rahimi M. (2023). Phytochemical screening and allelopathic potential
of phytoextracts of three invasive grass species. 13: 8080. DOI: 10.1038/s41598-
023-35253-x
Kiełtyka-Dadasiewicz A, Esteban J and Jabłońska-Trypuć A. (2024). Antiviral, antibacterial,
antifungal, and anticancer activity of plant materials derived from Cymbopogon
citratus (DC.) Stapf species. Pharmaceuticals (Basel, Switzerland). 17(6): 705.
DOI: 10.3390/ph17060705
Kumar S and Jena PK. (2014). Chromatographic, antibacterial and FT-IR analysis of
Dioscorea pentaphylla L. tuber extracts. Plant Science Research. 36(1& 2): 83-90.
Marndi S, Barik M and Kumar S. (2024). Phytochemical and cytotoxicity analysis of
Eulophia explanata Lindl. (Tuber extracts). African Journal of Biological Sciences.
6(4): 2751-2756.
Mety SS, Lal S, Topno SC, Rote K, Jena N, Kumar S and Behera B. (2024). Qualitative
phytochemical analysis of Solanum virginianum L. flower: a medicinal plant of
India. In: Plants and Secondary Metabolites, Volume 1. Ambika Prasad Research
Foundation, Odisha, India.
Mishra S and Bhatnagar S. (2024). Comparative analysis of methanol extract (leaf) of three
species of Sansevieria using chemical and biological parameters. World Journal of
Pharmaceutical Life Science. 10(2): 116-118.
Mukarram M, Choudhary S, Khan MA, Poltronieri P, Khan MMA, Ali J, Kurjak D and
Shahid M. (2021). Lemongrass essential oil components with antimicrobial and
anticancer activities. Antioxidants (Basel, Switzerland). 11(1): 20. DOI:
10.3390/antiox11010020
Nayak N, Bhujle RR, Nanje-Gowda NA, Chakraborty S, Siliveru K, Subbiah J and Brennan
C. (2024). Advances in the novel and green-assisted techniques for extraction of
95
Kapil et al., (2025). Plants and Secondary Metabolites, Volume 6; ISBN: 978-81-984754-0-4
bioactive compounds from millets: a comprehensive review. Heliyon. 10: e30921.
DOI: 10.1016/j.heliyon.2024.e30921
Panda PC and Panda S. (2012). Floral Diversity of Nandankanan Wildlife Sanctuary.
Nandankanan Biological Park, Forest and Environment Department Government
of Odisha, Bhubaneswar, Odisha, India.
Saxena HO and Brahmam M. (1996). The Flora of Orissa, Volume 4. Regional Research
Laboratory, Bhubaneswar and Orissa Forest Development Corporation Limited,
Bhubaneswar, Odisha, India.
Siddabathula N and Prasanna PV. (2023). Grasses of Telangana. Botanical Survey of India,
Kolkata, West Bengal, India.
Sinha MK, Dimri R, Sharma BP, Kumar S and Marndi S. (2023). Medicinal Grasses of India,
Volume 1. Ambika Prasad Research Foundation, Odisha, India.
Sujatha M, Johnson MA, Vanila D, Almeida RS, and Coutinho HDM. (2022). Biochemical
profile, in vitro toxicity, and cytotoxic activity of Eragrostis amabilis (L.) Wight.
Arn. And Eragrostis pilosa (L.) Beauve. Letters in Applied NanoBioscience. 11(2):
3480-3487.
Zeng Y, Pu X, Yang J, Du J, Yang X, Li X, Li L, Zhou Y and Yang T. (2018). Preventive and
therapeutic role of functional ingredients of barley grass for chronic diseases in
human beings. Oxidative Medicine and Cellular Longevity. 2018: 3232080. DOI:
10.1155/2018/3232080
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