POLYPHENOLIC CONTENT AND ANTIOXIDANT

ACTIVITY OF RIPE AND UNRIPE PAPAYA {Carica papaya

Linn (Caricaceae)} FRUIT PEELS

A
RESEARCH FINDING
BY
USMAN MUHAMMAD
NAS/15/BIO/1033

DEPARTMENT OF BIOLOGICAL SCIENCE, FACULTY OF NATURAL AND

APPLIED SCIENCES, SULE LAMIDO UNIVERSITY KAFIN-HAUSA. JIGAWA
STATE.

AUGUST, 2019
 INTRODUCTION
 Carica papaya (papaya) is a tree-like herbaceous plant, a
member of the small family Caricaceae and widely
cultivated for its edible fruit (Mohammed et al., 2014), By-
products derived from plants contain a variety of
phytochemicals and phenolic compounds such as
flavonoids, phenolic acids, saponin, tannins and others
(Jamal et al., 2017).
 Fruits peels are the major by-products obtained during the
processing of various fruits and some studies show that
these are good sources of polyphenols, carotenoids and
other bioactive compounds which possess various
beneficial effects on human health(Yario et al., 2004).
 CONT.
 Polyphenols are secondary metabolites characterized by
the presence of an aromatic ring bearing free hydroxyl
groups or engaged with a carbohydrate (Hanen et al.,
2012). These molecules are present in all plant parts and
are involved in many physiological processes such as cell
growth, root formation, seed germination and fruit ripening
(Ivanova et al., 2005).
 Antioxidant is any substance that when present at low
concentrations compared with that of an oxidizable
substrate significantly delays or inhibits oxidation of that
substrate (Anuj et al., 2016)
 STATEMENT OF RESEARCH
PROBLEMS
 Papaya releases a latex fluid when not quite ripe, which can
cause irritation and provoke allergic reaction in some people
which is due to lack of antioxidant activity .
 Excessive consumption of papaya can cause carotenemia, the
yellowing of soles and palms, which is otherwise harmless.
 Papain is also a potential allergen, according to Purdue
University, people who eat too much papaya and ingest high
levels of papain may develop symptoms consistent with hay
fever or asthma, including wheezing, breathing difficulties and
nasal congestion.
 JUSTIFICATION
 Traditionally peel of Carica papaya have been used
for treatment of a wide range of ailments, like in
treatment of malaria, dengue, jaundice and antiviral
activity (Arvind et al., 2013).
 Fruit peels of Carica papaya are rich in flavonoids,
tannin, saponin and phenolic compound (Dharma et
al., 1996, and Yario et al., 2004).
 polyphenols exhibit various biological activities such
as antioxidant, antifungal, antibacterial, antiviral
(Hanen et al., 2012).
 AIM AND OBJECTIVES
Aim of the study
 The study is aimed at determining and comparing
the polyphenolic contents and antioxidant activities
of the ripe and unripe Carica papaya fruit peels.
Objectives of the study
 To determine the polyphenolic contents of ripe and
unripe Carica papaya fruit peel.
 To determine the antioxidant activities of the ripe
and unripe Carica papaya fruit peel.
 MATERIALS AND METHODS
MATERIALS
 Folin ciocalteu reagent, Gallic acid, quercetin,
methanol, DPPH (2,2 diphenyl picryl
hydraxyl), ascorbic acid were collected from
departmental laboratory.
 All other chemicals, reagents and solvents are
also collected from laboratory, and were of
analytical grade.
 SAMPLE COLLECTION

 The plant materials (i.e. ripe and unripe Carica

papaya fruits) were purchased from Kafin-
Hausa market.
 SAMPLE PREPARATION
 Both the purchased ripe and unripe Carica
papaya was washed with tap water to remove
dirt. The peel was then removed from the
Carica papaya and allowed to shade dried for
about 3weeks, the shade dried peel was then
ground into a fine powder using laboratory
wooden pestle and mortar and was packed in
air tight containers and stored in a cool dried
place until required for use.
 SAMPLE EXTRACTION
 An extract was prepared using modified method used by
Vincent et al., (2015), 25g of the powdered Carica papaya
fruit peel was macerated in 500 ml of distilled water in a
conical flask. The content was mixed throughly and left to
stand for about 72 hours with an occasional shaking to
increase the extraction capacity. Thereafter, the soaked
peel was filtered with a muslin clothe and the resulting
aqueous crude was evaporated using a heating mantle to
obtain a powdered extract. The solid mark referred to as
the crude extract was stored in an air-tight experimental
bottle until ready for use.
 METHODOLOGY
TOTAL PHENOL
 The total phenolic content in the aqueous extract of ripe and unripe Carica
papaya fruit peels was determined spectrophotometrically with Folin
ciocalteu reagent using the modified method of Wolfe et al., (2003). An
aliquot of the extract (0.5 ml) was mixed with 2.5 ml of 10% Folin
ciocalteu reagent and 2 ml of Na2CO3 (7.5% w/v). The resulting mixture
was shaked for 15s and incubated at 40oc for 30 min for colour
development.
 The absorbance of the samples was measured at 765nm using UV visible
light spectrophotometer. The total phenolic content was expressed as mg/g
Gallic acid equivalent from the calibration curve using the equation
Y = 4.194x + 0.1713, R2 = 0.984
Where: Y was the absorbance and x was the Gallic acid equivalent
(mg/g)
The experiment was conducted in duplicate and the results are
expressed as ± SD values.
 TOTAL FLAVONOIDS
The method of Ordon et al., (2006) was used to estimate total
flavonoid contents of the extract solution based on the formation
of complex flavonoid–aluminum. A volume of 0.5 ml 2% AlCl 3
ethanol solution was added to 0.5 ml of extract solution. After on
hour of incubation at the room temperature, the absorbance was
measured at 420nm using UV visible light spectrophotometer. All
determinations were done in triplicate and values were calculated
from calibration curve obtained from quercetin using the equation:
Y = 4.405x, R2 = 0.990
Where Y was the absorbance and x was the quercetin
equivalent (mg/g).
 TOTAL FLAVONOLS
Total flavonol content was determined by adopting the
procedure described by Kumaran and Karunakaran (2007).
The reacting mixture consisted of 2.0 ml of the sample, 2.0 ml
of AlCl3 prepared in ethanol and 3.0 ml of (50 g/l) sodium
acetate solution. The absorbance was read 420 nm after 2.5
hour of incubation at 20oc. Total flavonol content was
calculated as quercetin (mg/g) equivalent from the calibration
curve using the equation:
Y = 4.405x, R2 = 0.990
Where Y was the absorbance and x was the quercetin
equivalent (mg/g).
DPPH radical scavenging activity
The method of Liyana–pathiranan and Shahidi (2005) was used for the
determination of scavenging activity of DPPH free radical in the extract
solution. A solution of 0.135 mM DPPH in methanol was prepared and
1.0 ml of this solution was mixed with 1.0 ml of extract prepared in
methanol containing 0.025 – 0.5 mg of the plant extracts and standard
drug separately (ascorbic acid). The resulting mixture was shake
thoroughly and left in the dark at room temperature for 30 min. the
absorbance of the mixture was measured spectrophotometrically at 517
nm. The ability of the plant extract to scavenge DPPH radical was
calculated by the equation:
DPPH radical scavenging activity = {(Abscontrol – Abssample)/
(Abscontrol)} x 100
Where: Abscontrol is the absorbance of DPPH radical + methanol
Abssample is the absorbance of DPPH radical + extract or standard.
 DATA ANALYSIS
 All the analysis were performed in duplicate
and the result were presented in a bar chart.
 RESULT
Table 1. percentage yield of extract recovered

EXTRACT YIELD% COLOUR

RIPE 34.64 Dark brown
UNRIPE 32.88 Dark brown
 120

100

80
Content

60 phenolic mgGAE/g
flavonoid mgQUE/g
40 flavonol mgQUE/g

20

0
ripe unripe
C. papaya sample

Fig. 1 variation in polyphenolic content in ripe and

unripe Carica papaya fruit peel.
% of radical scavenging activity 120

100

80

60 RSA OF STD
RSA OF RIPE
40 RSA OF UNRIPE

20

0
0.03125 0.0625 0.125 0.25 0.5
Concentration mg/ml

Fig. 2 variation in scavenging activity of

Carica papaya fruit peel
 DISCUSSION

 After extraction unripe Carica papaya fruit peel found to

have higher percentage yield of extract.
 This result shows that the unripe Carica papaya fruit peel
has higher TPC.
 This study is similar to study carried by Maisarah et aI.,
(2013), unripe Carica papaya fruit peel have TPC of 94.89
± 0.22 mg GAE/ 100 g and ripe Carica papaya fruit peel
have 30.32 ± 6.90 mg GAE/ 100 g dry weight and TFC
unripe Carica papaya fruit peel have TFC of 333.14 ± 1.03
and 53.44 ± 6.64 and mg rutin equivalent/ 100 g of ripe
Carica papaya fruit peel.
 The result from Govindarajan et al., (2007)
shows that the flavonol content of unripe
extract was 42.89 ± 0.22 mg GAE/ 100 g and
ripe extract was 30.32 ± 6.90 mg GAE/ 100 g.
 All the result from the above mentioned
researchers shows that the unripe Carica
papaya fruit peel possessed higher
polyphenolic content than the ripe peel.
 This variations in the polyphenolic contents
are due to stage of ripening.
 CONCLUSION
 The present result demonstrate that the aqueous
extract of unripe Carica papaya fruit peel possessed
high phenolic content, flavonoid content, flavonols
contents and antioxidant activity than the ripe Carica
papaya fruit peel.
 RECOMMENDATION
 The use of unripe carica papaya peel is recommended as it
possess antioxidant activities.
 Unripe carica papaya peels is of important, as such is should not
be discarded
 Further studies is recommended to evaluate the other contents of
ripe and unripe carica papaya fruit.
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