PHYTOCHEMICAL SCREENING, ANTIMICROBIAL AND

CYTOTOXICITY PROPERTIES OF PIKAW (COLOCASIA SP.

CF. FORMOSANA HAYATA)

Prepare by: Juliet D. Panuncial

 ABSTRACT
The phytochemical, antibacterial, antifungal, and
cytotoxic properties of the ethanol extract of Pikaw
(Colocasia sp. cf. formosana Hayata) were conducted in
this study. These were appropriately conducted in two
accredited science laboratories, the Department of
Science and Technology in Tuguegarao City, Cagayan
and Center for Natural Sciences, Saint Mary‘s University in
Bayombong, Nueva Vizcaya.
 There are favorable results of the phytochemical
screening, bacterial assay, fungal assay and cytotoxicity
assay of the Pikaw ethanolic extract. Pikaw has
phythochemicals that include flavonoids, tannins,
saponins, essential oil, triterpenes, fatty acids, sugar,
coumarins, anthrones, phenols, alkaloids, steroids and
anthraquinones
 In addition, pikaw ethanolic extract cannot inhibit
bacteria S. aureus, E. coli and B. subtilis but it has high
ability to inhibit the fungus C. albicans. The range of the
zones of inhibition of Pikaw ethanolic extract on Candida
albicans is 29-31. This range is comparable with
miconazole, cltrimazole and ketoconazole. Hence, the
Pikaw ethanolic extract can be made into products to
serve as substitute of commercially available antifungal
diseases caused by Candida albicans.
 Pikaw has also a cytotoxic property because
after 18 hours the LC50 = 941.528 ppm, after 21 hours
the LC 50 = 743.894 ppm and after 24 hours the
LC50=634.807 ppm.
Recommendations include the (1) preparation
antifungal cream, ointment and other antifungal
products made out of pikaw ethanolic extract that
resemble the commercial preparations of
miconazole, clotrimazole and ketoconazole; and (2)
isolation the flavonoids since this has a role on the
cytotoxic property of the pikaw ethanolic extract
then retest cytotoxicity.
 INTRODUCTION
Background of the Study
Nueva Vizcaya is blessed with flora from Diadi to
Alfonso Castañeda. Its forest areas have hidden water falls
that contribute to its cool climate. Where perennial streams
or springs were located at the edge of forest, along
roadsides is a morphologically distinct wild form or species
of taro (Colocasia sp. cf. formosana Hayata) growing on
stream banks, and among rocks next to small waterfalls.
This wild taro is popularly known as the edible pikaw
among Ilocano.

But there are many species of wild taro. Pikaw is just

one of them.
Matthews, et al., (2007) revealed that to understand the
origins and history of a crop, it is necessary to consider
where wild populations of the same species were naturally
distributed before people began interacting with them
and using them. Ethnographic observation of how people
use wild populations can also help by suggesting how wild
plants were taken into cultivation, and how they were
domesticated.
 Domestication, or the genetic modification of a
useful species, requires selection from a variable
population, and isolation of selected forms from the
dominant genetic influence of a wild gene pool. Their
survey has not proven the existence of natural, indigenous
wild taro (C.esculenta) in the Philippines; wild taro is
abundant in some areas, whether or not breeding
populations are present. Wild taro is economically
significant, and is subject to loose management as a
community resource. Priorities for future research are on
the natural and cultural history of taro in the Philippines.
 In the mountainous Ifugao region, in northern
Luzon, wild taro did not appear abundant. This might
be due to the much dissected nature of the terrain,
which made access difficult, and the lack of large
open habitats that are not closely controlled and
cultivated. It is also possible that the cooler climate of
this northern mountain region is not ideal for wild taro.
On the dryslopes of Mt Arayat, further south, and in
western Luzon, wild taro was completely absent. By
contrast, wild taro was very abundant in open, wet,
and humid areas, and at middle altitudes on Mt
Banahaw and Mt Apo.
Possible source populations may or may not be located at
higher altitudes on these mountains, in less-disturbed and
more inaccessible forest areas. The most common
phenotype seen, from Ifugao to Mt Banahaw and Mt Apo,
displayed green leaves, with white lower parts, relatively
small corms, and long stolons. In all obvious respects, this is
like the wild type taro present in northern Australia, Papua
New Guinea, Indonesia, Myanmar, and Vietnam—all
areas where flowering and breeding populations are
present (observations by Matthews, and others).
 As noted in the introduction, Brown (1920) did
not mention wild taro in his list of wild food plants of the
Philippines. He did indicate that ̳personal judgment‘
was used in making the list, and that: ̳Many people
may consider some of those (plants) treated in the
present list as worthless, and some common species
which are omitted as good‘, and that his list
was ̳probably very far from complete‘. In later
publications, Brown (1941) and Monsalud et al. (1966)
mentioned the use of other wild aroids, including
species that are also cultivated, but made no mention
of taro as a wild plant.
From their own observations, and those of their local
informants, it is clear that wild taro is widely used and
popular as a vegetable food, mainly for the leaves, but
also to some extent for the stolons, and as a source of
leaves for pig fodder.
Cordero-Fernando (1992) stated that people in the Bikol
region (southern Luzon) cook and eat only the leaves of
the gabi plant, and that most gabi grow wild. In the past,
wild taro may have been important as a source of starch
in times of hunger, and as a free and nutritious leaf
vegetable at all times, especially among rural
households.
 Taro has been focused on cultivars that are grown
primarily for their corms. Corms may have more nutritional
or economic value, as a starchy food and for trade, in
most social contexts. Pardales and Villanueva (1984) did
not mention leaf varieties in their report on field trials with
Philippine and Hawaiian cultivars at Baybay in Leyte.
Lebot et al. (2004) surveyed the corm qualities of 2,298
taro accessions collected in Asia (including the
Philippines) and the Pacific, and also did not comment on
the use of leaves. In Vietnam, a national collection of 350
taro accessions included approximately 25% with edible
petioles or young leaves (Nguyen, 2000). Wild and
cultivated taros with edible leaves are likely to be under-
represented in national collections of the Philippines and
most other countries.
To encourage further research on edible- leaf forms
of taro in the Philippines, a collection of several
distinct forms of wild taro in the vicinity of Mt.
Banahaw, and transferred them to a living taro
collection maintained by the College of Agriculture,
University of Los Baños.
 In the study of Adaoag, pikaw is under
classification Araceae. It is a herbaceous aroid. It has
an underground fleshy stem or corm that extends to a
large cylindrical stalk, and wide ovate leaves. The
leaves are cuneate-based and acute-apexed.
Clusters of naked female flowers are in a compact
mass on the basal part of a fleshy stalk (spadix) give
rise to berries. On the same stalk are the male and
neutral flowers protected by a hood, the spathe. The
fruits are in triangular tube containing the berries
inside.
 Pikaw is documented in the highland municipalities
of Abra like Boliney, Sallapadan Lacub, Malibcong,
Daguiman and Bucloc along river systems and streams at
higher altitudes. It anchors its roots in between and
among rocks and well adapted to sandy clay soil with
good drainage and high organic matter.
The plant contains a sap that causes skin irritations or
itchiness found concentrated in the flowers and which
gives a permanent black stain to clothes. Corms are not
gathered by local
folks as they are organs for plant horizontal growth and
food reservoir for the plant. The phytochemical analysis
results show the plant contains gums, mucilages,
glycosides, carbohydrates, reducing sugars, tannins and
derivatives, proteins and derivatives, flavonoids,
deoxysugars, unsaturated sterols and triterpenes,
polyphenolic compounds Adaoag.
 The young leaves are the edible portions leaving
the corms for further growth. These parts are hashed and
bundled piled into the cooking pot. They are treated with
fish sauce, onions, garlic, vinegar, and ginger and
sometimes with coconut milk, topped with meat,
sardines, or dried fish. It is best not to disturb/stir pot while
cooking so the calcium oxalate crystals in the sap will be
thoroughly cooked.
 Philippines has different epidemics that people
suffer and that the researchers want to help all the people
that will be affected of diseases and after knowing about
the plant Pikaw, the researchers became curious about its
chemical properties that can help not just Ilocanos but
also other people needed to be cured. This reason made
the researchers to continue and aim for the solutions that
can help all people who are in need. The researchers
aimed to know all the properties of the plant that can
treat diseases.
 It inspired the researchers to continue their journey to
know about the plant and how can it help people.
Ilocanos kept on eating Pikaw and the researchers
want to tell them that the plant that they eat is one of
the possible cures for their sickness. All the possible
effects of its chemical properties of Pikaw it can be
cure of their sickness. The researches always kept in
mind that if they prove that Pikaw is an effective plant
they can help all the people who are suffering in all
there sickness.
Pikaw is one of the many species of wild taro
Significance of the Study
This study would benefit the research community
most especially the consumers of this plant in the upland
areas of Nueva Vizcaya. This study can help the people to
understand the properties of Pikaw and what can it do for
their health. There are some chemical properties of the
plant that can treat some illnesses.
Scope and Limitations of the Study
The study will be confined on the phytochemical
screening to know the chemical properties of pikaw;
antimicrobial to know if it is possible to kill bacteria and
fungi and cytotoxic properties to know if it is possible to kill
cells. This study does not know if any attempt has been
made to assess the commercial value of wild taro.
 METHODOLOGY
Procedural Flowchart

PREPATION OF THE CRUDE EXTRACT

 Gathering of Pikaw ( Colocasia sp.cf. formosana Hayata )

Air-drying

Pulverization

Extraction

Filtration

Water Bath

Phytochemical Screening Antimicrobial Assay Cytotoxicity Assay

PHYTOCHEMICAL SCREENING
 Phytochemical Screening

Capillary Tube Spotting

Cromatography

Preliminary Methanolic Dragendroff’s

Test Pottasium Hydroxide Reagent

REAGENTS Vanillin Magnesium Ninhydrin

Sulfuric Acid Acetate

Naphthol- Antimony Potassium Ferricy

Sulfur Acid ( III ) Chloride ferric Chloride
ANTIMICROBIAL ASSAY
 Antimicrobial Assay

Preparation of Culture
Media

Inoculation Procedure

Preparation of Culture
Media

Streak Plate Procedure Streaking Patterns

Materials
• 1 Kilogram of fresh pikaw leaves for air-drying
• All standards materials for phytochemical screening,
antimicrobial assay, and cytotoxicity study obtain from a
standard laboratory ( DOST, SMU)
Methods/Procedures
Phytochemical screening:
Phytochemical examination were carried out for all the
extracts as per the standard methods.
Reagent used:
1. Preliminary Test. (Essential oils). Heat at 90°C; violet spot under UV
365 nm.
2. Vanillin Sulfuric Acid. (Higher alcohols, steroids, triterpenes,
essential oils, phenols, fatty acids). Heat at 90°C (char). Triterpenes
and sterols appear mainly as blue violet spots under long wave UV
365 nm. Essential oils from zones with wide range of colors under long
wave UV 365 nm. Phenols appear as brown spot under visible light.
Fatty
acids as yellow spot under visible light.
3. Napthol-sulfuric Acid. (Sugars). Heat at 90°C (char). Blue dark spot
under visible light.
4. Methanolic Potassium Hydroxide (KOH-MetOH). (Anthraquinones,
coumarins,
anthrones). Anthraquinones give orange coloration under visible
light. Coumarins react to form blue colored zone under long wave
UV 365 nm. Anthrones give yellow zones long wave UV 365 nm.
5. Potassium Ferricyanide-ferric Chloride. (Tannins,
flavonoids, phenols). Blue spots under visible light.
6. Dragendroff’s Reagent. (Alkaloids). Brown-orange
visible spots immediately upon immersing into test
reagent; colors are not stable.
7. Antimony (III) Chloride. (Flavonoids, steroids). Intense
yellow to orange upon immersing for glycoside
flavonoids; fluorescent colors under long wave UV 365 nm
for steroids.
8. Magnesium Acetate. (Anthraquinones). Heat at 90°C
(char). Orange-violet color after heating at 90°C.
9. Ninhydrin. (Amino Acids). Violet spot upon dipping.
RESULTS AND DISCUSSION
The DOST and CNS Lab Results on Phytochemical
Screening of Pikaw
Phytochemical analyses were conducted at the Saint
Mary‘s University-Center for Natural Sciences of
Bayombong and DOST Laboratory Testing in Tuguegarao.
This was done to establish validity and reliability of results.
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Table 1: Phytochemical Components of Pikaw

Phytochemical SMU Lab Result DOST Lab Result
Flavonoids Possitive Possitive
Tannins Possitive Possitive
Saponins Possitive
Essential Oil Possitive Not Tested
Triterpenes Possitive Not Tested
Fatty Acids Possitive Not Tested
Sugar Possitive Not Teted
Coumarins Possitive Not Tested
Anthrones Possitive Not Tested
Phenols Possitive Not Tested
Alkaloids Possitive Not Tested
Steroids Possitive Not Tested
Anthhrauinones Possitive Not Tested
 At SMU-CNS the following phytochemicals were
found: essential oils, triterpenes, fatty acids, sugar,
coumarins, athrones, tannins, flavonoids, phenols,
alkaloids, steroids and anthraquinones. At DOST Lab,
the following phytochemicals were found: flavonoids,
saponins and tannins.
In both labs, the following were found: essential
oils, triterpenes, fatty acids, sugar, coumarins, athrones,
tannins, flavonoids, phenols, alkaloids, steroids,
anthraquinones and saponins.
 CONCLUSIONS AND RECOMMENDATIONS

There are favorable results of the

phytochemical screening, bacterial assay, fungal
assay and cytotoxicity assay of the Pikaw ethanolic
extract. Pikaw has phythochemicals that include:
Flavonoids, Tannins, Saponins, Essential Oil,
Triterpenes, Fatty Acids, Sugar, Coumarins, Anthrones,
Phenols, Alkaloids, Steroids and Anthraquinones
 Pikaw cannot inhibit these bacteria, namely: S.
aureus, E. coli and B. subtilis but it has high ability to
inhibit the fungus named C. albicans.

The range of the zones of inhibition of Pikaw

ethanolic extract on Candida albicans is 29-31. This
range is comparable with miconazole, cltrimazole
and ketoconazole but not with fluconazole. Hence,
the Pikaw ethanolic extract can be made into
products to serve as substitute of commercially
available antifungal diseases caused by Candida
albicans.
 Pikaw has also a cytotoxic property because
after 18 hours the LC50 = 941.528 ppm, after 21 hours
the LC 50 = 743.894 ppm and after 24 hours the
LC50=634.807 ppm.

The researchers recommend to (1) prepare

antifungal cream, ointment and other antifungal
products made out of pikaw ethanolic extract that
resemble the commercial preparations of
miconazole, clotrimazole and ketoconazole; and (2)
isolate the flavonoids since this has a role on the
cytotoxic property of the pikaw ethanolic extract
then retest cytotoxicity.