EXTRACTION AND SCREENING OF

PHYTOCHEMICALS FROM THE

LEAVES OF SELECTED
FERN SPECIES

Gabriel Angelo Celebria

Raphael Yturralde
Bryan Atas
 Phytochemicals
• Naturally occurring, non-nutritive plant
chemicals

• any chemical, nutrient, or antioxidant that

contain protective, disease preventing
compounds derived from a plant source

• Determines the medicinal value of the

plant
 Ferns

• Spore-forming plant of the family Pteridophyta

• Used as ornamental plants, vegetable, herbal

medicines, building materials
 Significance of the study
Results would provide information

• regarding the presence of phytochemicals

in ferns

• Types of phytochemicals in each fern

species
 Statement of the problem
• Do the leaf extracts of ten species of
ferns contain alkaloids, flavonoids,
saponin, and tannins?
 Objectives of the study
• To collect leaf samples from ten selected
species of ferns.

• To obtain extracts from the leaves of ten

selected species of ferns.

• To determine the presence of alkaloids,

flavonoids, saponins and tannins in the
leaf extracts of ferns.
• Alkaloids
• Flavonoids
• Saponins
• Tannins
Our study will be concerned with only four
types of phytochemicals
 Alkaloids
• usually containing at least one nitrogen
atom in a heterocyclic ring

• Protects plants from herbivores

» McKey(1974)
e.g. ephedrine

• anti-cholinergenic, anti-inflammatory,
anti-spasmodic, hyperglycemic, sedative,
tranquilizer and vasodilator
» (www.shaman-australis.com/~auxin/heimia.html).
 Flavonoid
• class of plant secondary metabolite based
around a phenylbenzopyrone

• produce pigmentation on plants

e.g. flavone
• protect against UV-B radiation in apples
» (Solovhcenko A. and Schimitz-Eilberger M. 2003)

• anti-allergic, anti-inflammatory, anti-microbial,

anti-oxidative and anti-cancer activity
 Saponins
• glycosides of steroids, steroid alkaloids or
triterpenes found in plants

• form a waxy protective coating on plant skins

• protect against cancers, decrease blood

lipids, lower blood glucose response, inhibit
dental caries, inhibit platelet aggregation,
anti-microbial, and lowers cholesterol
» (www.eas.com/glosary/glossary.asp?glos_pk=278)
 Tannins
• astringent, bitter-tasting plant polyphenols
that bind and precipitate proteins
Hydrolysable
• Anti-microbial Condensed
•• Divided
Have polyol
into K and others• 2006)
hydrolysable
» (Yao polymers of 2 to 50
and condensed
carbohydrate (or more) flavonoid
molecule at units that are joined
• the
defend
centerthe plants against herbivores by
by carbon-carbon
decreasing protein availability
bonds
• Not susceptible to
hydrolysis
 Ferns
• common name of any cryptogamous or
spore-forming plants.

• Belongs to the family Pteridophyta

 Ferns used in the study
• Asplenium nidus • M. punctatum ‘Grandiceps’

• A. musifolium • Nephrolepis exaltata

• Davallia solida • N. biserrata

• Drynaria quercifolia • N. hirsutula

• Microsorium • Phlebodium aureum

heterocarpum
Asplenium nidus A. musifolium
• epiphyte • epiphyte

• leaves are simple, • leaves are simple,

stiff, oblanceolate, stiff, oblanceolate,
light green, wavy and light green,
margins with shiny crowded
surface.
Davallia solida Drynaria quercifolia
• epiphytic, dimorphic • epiphytic fern, with 2
fern kinds of leaves
– Sterile leaves are
pinnate, sub opposite, – nest leaves which are
broadly deltoid long concave, resembling
– Fertile leaves are the oak-leaf
lobed, with each lobe – foliage leaves, used
bearing several for photosynthesis
sporangia.

• used in haemoptysis
and is anti-helminthic
Microsorium M. punctatum
heterocarpum ‘Grandiceps’
• short creeping • leaves are bright
rhizomes green, elongated,
• simple, narrowly more or less ruffled at
decurent almost to the margin, and
the base, and ovate- forking in various
lanceolate fronds segments at the apex.

Phlebodium aureum
• a creeping fern with thick rhizomes covered
with rusty brown hair like scales
Nephrolepis biserrata N. hirsutula
• sori pinnae are about • leaflets are variably
5cm long, fresh spaced ,linear-
green, leathery at the lanceolate.
ends of the veins • sori are dot like and
are near the margins.

N. exaltata
• leaflets are overlapping with each other and
are tiny
Methodology
 Preparation and collection of samples

Preparation of extracts

Flavonoid screening

Tannin screening
Alkaloid analysis

Saponin screening
 Preparation and collection of
samples
• All ferns (except Drynaria quercifolia) were
taken from WVSU

• Drynaria quercifolia was taken from

Mandurriao

• Additional leaves of A. nidus and N.

bisserata were taken from PSHS-WV
 Flavonoid screening
Flavonoid screening

Bate-Smith and Metcalf Wilstatter “cyanidin” test

test for test for y-benzopyrone

leucoanthocyanins nucleus
Alkaloid screening
Alkaloid screening (fresh)
 Saponin screening
•Libermann – Buchard Test
 Tannin screening
Test tube method

Ferric chloride test Gelatin test

Results and Discussion
 Table 1. Presence and absence of alkaloids, flavonoids,
leucoanthocyanins, triterpenoid saponin,
condensed tannin and hydrolyzable Results
tannin in the leaves selected fern species.
flavonoids tannin

alkaloid Bate- saponin

Fern species Wilstatter Ferric
smith & gelatin
Cyanidin chloride
Metcalf
A. musifolium positive positive positive negative positive Both*

A. nidus positive positive positive positive positive Both*

D. solida positive negative negative positive positive Both*
D. quercifolia positive positive positive positive positive condensed
M. heterocarpum positive positive positive positive positive condensed
M. punctatum positive positive positive positive positive Both*
N. bisserata positive positive positive negative positive Both*
N. exaltata positive positive positive positive positive Both*
N. hirsutula positive positive positive positive positive Both*
P. aureum positive positive positive positive positive Both*
*both- condensed and hydrolyzable tannins are present
 Discussion
• Phytochemicals are present in the plants
structures because they provide defensive
capabilities to plants.
 Alkaloid
• Alkaloids provide plants with defense against
herbivores,
Fern species viruses, bacteria and fungi.
alkaloid

A. musifolium positive

•A. nidus
alkaloids are concentrated
positive in the plant parts
that are most susceptible
D. solida positive to attacks, such as
D. quercifolia positive
leaves
M. heterocarpum
and stems
positive
» McKey (1974).
M. punctatum positive
N. bisserata positive
N. exaltata positive
N. hirsutula positive
P. aureum positive
 Flavonoids
flavonoids

Fern species Bate-smith & Wilstatter

Metcalf Cyanidin

A. musifolium positive positive

A. nidus positive positive

D. solida negative negative
D. quercifolia positive positive
M. heterocarpum positive positive
M. punctatum positive positive
N. bisserata positive positive
N. exaltata positive positive
N. hirsutula positive positive
P. aureum positive positive
 Flavonoids
• are present in all the leaves of selected
ferns except the giant hare’s foot fern (D.
solida)
• D. solida usually grows in places near bodies of
water and away from direct sunlight.

• protect plants from UV-B radiation

» (Solovchenko A. and Schimitz-Eilberger 2003)
 Saponins
saponin
Fern species
A. musifolium negative

A. nidus positive

D. solida positive

D. quercifolia positive

M. heterocarpum positive

M. punctatum positive

N. bisserata negative

N. exaltata positive

N. hirsutula positive

P. aureum positive
 Saponin
• Saponin is one of the most common
phytochemical in plants
» (Cornel University 2004).

• Only the leaves of sword fern (N.

biserrata) and the dwarf bird’s nest fern
(A. musifolium) lack saponins
 Tannin
tannin

Fern species Ferric

gelatin
chloride
A. musifolium positive Both*

A. nidus positive Both*

D. solida positive Both*
D. quercifolia positive condensed
M. heterocarpum positive condensed
M. punctatum positive Both*
N. bisserata positive Both*
N. exaltata positive Both*
N. hirsutula positive Both*
P. aureum positive Both*
 Tannins
• protect plants from ruminants or
herbivores
» (Robins C. 1987).

• All leaf samples contain tannin

Summary, Conclusion and
Recommendation
 Summary
• The tests show that 7 of 10 ferns show the
presence of all the phytochemicals tested
in this study; specifically, alkaloids,
flavonoids, saponins and tannins. At most
one phytochemical is absent from the fern
leaves.
 Conclusion
• Most of the fern species contain the tested
phytochemicals in their leaves.

• Of the ten fern species tested, only three

lacked one type phytochemical.
• All ferns contain alkaloids and tannins in their
leaves; which, implies that the ferns have a
potential medicinal value.

• The presence of flavonoids in all fern species,

except the hare’s foot fern, implies that most of
the fern leaves have anti-oxidative properties.

• The presence of saponin in all of the tested fern

leaves, except for A. musifolium and N.
bisserata, suggest that eight out of ten tested
fern species have anti-microbial properties in
their leaves.
 Recommendation

• Further test regarding it should be

performed
– Anti-microbial
– Anti-oxidative
– Anti-cancer
– quantification
 Alkaloid screening
• N. hirsutula
alkaloid screening

• showing the
formation of a
white precipitate
 Flavonoid

• Flavonoid screening showing Bate-Smith

Metcalf and Wilstatter “cyanidin’’ test on the
extracts of Phelbodium aureum
 Saponin
• Saponin screening
showing the darker
control on the right

• A. nidus
 Tannin

• Ferric chloride test, showing the tannic

acid on the center
 END
Thank You