Antioxidant component of Coconut haustorium

1. FUNCTIONAL COMPONENTS OF THE COCONUT HAUSTORIUM (Cocos
nucifera)
TIMOTHY M. ZAGADA
B.S. Nutrition Student, College of Human Ecology, University of the Philippines, Los Baños College,
Laguna 4031 Philippines
Antioxidant activity in foods has been suggested to have protective effects against
chronic diseases in humans. This paper gives information about the functionality of the
antioxidants that are found in the Coconut haustorium (Cocos nucifera) using the method
suggested by the Institute of Plant Breeding Manual by Rodriguez et.al., The mean (all in
mg/100g ) of total phenols was determined with 102.87, Flavonoids with 167.88, and Saponins
with the largest observed mean 8051.86 and the total antioxidant activity has a mean of
89.36 . Results show that phenols deviate by 17.34, flavonoids by 18.43 and saponins by
702.5. The antioxidant activity also deviated from its mean only by 2.99 or only 10% (CV). No
tannins and alkaloids were detected.
The results of the functional components were correlated with the antioxidant
activity by computation of the correlation coefficient (r). Phenols obtain an r value of 0.48,
flavonoids with 0.68 and the highest was saponins with 0.81 which has the strongest
relationship to the antioxidant activity.
INTRODUCTION
The tenet "Let food be thy medicine and
medicine be thy food," espoused by Hippocrates
nearly 2,500 years ago, is receiving renewed
interest. In particular, there has been an
explosion of consumer interest in the health
enhancing role of specific foods or
physiologically-active food components, so-
called functional foods, according to their
generally accepted definition, are "any food or
food ingredient that may provide a health benefit
beyond the traditional nutrients it contains"
(Hasler, 1998).
Antioxidants are compounds that protect
cells against the damaging effects of reactive
oxygen species, such as singlet oxygen,
superoxide, peroxyl radicals, hydroxyl radicals
and peroxynitrite (Buhler, 1962). Antioxidants
may be considered functional by virtue of having
potential for such actions as anti cancer and anti
cardiovascular-disease effects or being modifiers
of oxidative damage and defence mechanisms
(Fore, 2000).
Fruits and vegetables are good sources of
antioxidants other than vitamin C and E, B-
Carotene. Dietary intake of fruits, vegetables and
their constituents, which are potent effectors of
biological systems in humans, has been
suggested to have protective effects against
chronic diseases (Agarwal & Rao, 2000).
The Coconut (Cocos nucifera) haustorium is
the embryo plant grows into the inside of the nut
to make an organ of spongy tissue ("apple" or
"sucker"). It commences to develop at the start of
germination when the shoot or roots have not yet
emerged through the husk. It takes about 4
months to expand to almost fill the cavity of the
nut, except in very large nuts. It transfers
nutrients from the endosperm to the young
seedling. ((IFAS), 2000). Since the “apple” is
slightly sweet and slightly oily with a cotton
wool-like texture, it is usually placed in one side
for eating. In places where coconuts grow,
children walking to school may grasp the leaves
of a sprouted seednut and uproot it. Still holding
it by the leaves, they swing it against the trunk of
the nearest mature palm to split the husk and
crack open the shell. Then they pick out and eat
the “apple” (Harries, 1994). The Haustorium is
used as a major ingredient of the “Minatamis na

2. Functional Components of the Coconut Haustorium
Tumbong” and is also cooked as a vegetable
(Martin 1999). The Haustorium is also shown to
possess many active enzymes; among them are
cellulases, lipases and proteinases.
MATERIALS AND METHODS
Collection of samples
A geminating coconut seed from the local market
of Los Baños was obtained with the aid of the
coconut vendor to determine the presence of the
Haustorium. The Coconut was free from
parasites and pests with the husk firm and nut
punctured.
Preparation of Samples
The germinating coconut seed is cut
into half using a bolo without damaging the
haustorium. It was then separated from the seed
by slowly wedging it with a spoon. The
Haustorium was chopped and diced as
preparation for the masceration process.
Treatment and Variables
In the extraction process, five grams of
the haustorium was needed per replicate then
placed inside a mortar. Twenty ml of methanol
was then added and was mascerated for 15
minutes. The mixture was filtered to gain an
extract about 5-8 m to be placed in vials. The
extraction process was repeated six times
representing the number of trials or replicates.
The functional components; phenols, flavonoids,
tannins, saponins, and alkaloids served as the
dependent variable and the independent variable
were the samples or replicates.
Chemical Analyses
Procedure of the chemical analysis
followed the method suggested by the Institute
of Plant Breeding Manual by Rodriguez et.al with
slight modifications.
Total Phenols
1. Obtain 0.20 ml of the extract
2. Add 2.80 ml of distilled water
3. Add 1ml of 0.2M Na2CO3
4. Add 0.20 ml of Folin Ciocalteus reagent
5. Heat in boiling H20 bath for 15 min.
6. Cool to room temperature.
7. Get Trans reading at 710nm.
Flavonoid
1. Get o.50ml of the xtract
2. Add 4.50ml of distilled H20
3. Add 0.20ml of F-C reagent.
4. Add 0.50ml of saturated Na2CO3
5. Vortex mix for 15 seconds
6. Add 4.3ml of distilled H2O.
7. Mix and let to stand at room temperature for
an hour.
8. Get Trans reading at 725nm
Tannins
1. Obtain 0.5ml of the extract.
2. Add 3.0ml of 1% Vanillin reagent.
3. Stand at room temperature for 20 minutes.
4. Get Trans reading at 500nm.
Saponins
1. Do the extraction process with ethanol as the
solvent.
2. Pass 2.5ml of the extract in the PVPP
column (Polyvinylpolypyrollidone) which
was soaked in distilled water overnight.
3. Get 0.50ml of the passed extract and put in a
separate test tube.
4. Add 0.50 ml of acetic HAC to the extract.
5. Add 3.0ml of L-B reagent (Lieberman-
Buchard).
6. Heat at 90-100C for 30 minutes.
7. Cool to room temperature.
8. Get Trans Reading at 450nm.
Alkaloids
1. Get 2ml of the extract.
2. Add 10ml of 1% Hydrochloric acid
3. Add 5.0 ml of concentrated ammonia and
transfer into a 50-ml separatory funnel.
4. Extract thrice with CHCL3 at 10ml each
time.
5. Combine extract and pass in a sodium
sulfate column.
6. Evaporate extracts.
7. Add 2.0ml 1% HCL and 1.0ml CHCL3.
Antioxidant Activity (AOA)
1. Get 2ml of extract.
2. Add 2ml of phosphate buffer.
3. Add 2ml KFe(CN)6 and put in water bath at
50C for 20 minutes.
4. Add 2.5ml of 10% TCA.
5. Add 2.0ml of distilled water.
6. Add 0.5ml of FeCl3
7. Get Trans reading at 500nm.

3. Functional Components of the Coconut Haustorium
RESULTS AND DISCUSSION
A. Functional components and AOA of the
coconut haustorium.
Table 1.Functional components and AOA of the
coconut haustorium
The mean phenolic content of the
coconut Haustorium is 102.87 mg/100g which is
relatively higher compared to other fruits like
watermelon (64), cherry (41), pineapple (67) and
is relatively closer to avocado (101). All were
based on the study by Vinson, et.al entitled
“Phenol Antioxidant Quantity and Quality in
Foods: Fruits”. According to BRAVO (1998),
the presence of polyphenols in plant foods is
greatly influenced by genetic factors,
environmental conditions, degree of ripeness,
variety, etc.
Flavonoid was also present and the
obtained mean value was 167.88mg/100g.
According to Torel (1986), The antioxidant
efficiency of Flavonoids depends on: a) their
concentration wherein an increase in the
concentration from 1 to 100ug/ml, increases the
antioxidant efficiency of Flavonoids, and b) the
degree of saturation of the fatty acid such that an
increase in the unsaturation results in a decrease
of the antioxidant efficiency.
The value obtained from saponins was
702.50mg/100g. Coconut oil is used in the
manufacture of soaps, detergents, and shampoos
because it has high levels of lauric acid, an
ingredient that gives soap a quick-lathering
property and also due to its hardness, ready
stability, and free leathering properties of the
sodium soaps made with it (Technonet, 1990). In
the study conducted by Tiocson (2006), it was
determined that the Haustorium contains large
amounts of Lauric acid. According to Martin
(1999) the haustorium is an active site of enzyme
secretion, allowing the ready absorption of the
flesh including its oil reserves thus; it is
manifested by the high saponins reading in the
experiments’ results.
No observations were accounted for
tannins and alkaloids which was manifested by
the absence of turbidity during the experiment.
According to Hagerman (2002), tannins have an
astringent sensation which was not reflected by
the coconut Haustorium.
B. Standard Deviation and the Correlation of
the Funtional Components to the Antioxidant
Activity.
Table 2. The mean, standard deviation and correlation
coefficient.
The standard deviation measures the
average deviation or the average distance of the
observations from the mean of the data set. It is
also a measure of dispersion and variability of
the data. A small standard deviation in the
experiment means that the results are close to the
mean of the data which can be expressed in
terms of accuracy or consistency of the results.
The standard deviation for phenols was 17.34
which means that the result of the experiment
deviates by 17.34 (102.87±17.34). The standard
deviation computed for Flavonoids was 18.43
wherein the experiments’ results also deviate
with a value of 18.43 (167.88±18.43).
The saponins exhibits the most
deviation wherein there is 702.5 average distance
from the mean (8051.86±702.5). The standard
deviation becomes larger as the observations
become more dispersed from the mean. The
MAX and MIN values for the data obtained from
the saponins analysis was 8814.39mg/100g and
7255.23mg/100g with a computed Range of
1559.16. Since all observations contribute to the
computation, the higher the value of the
observation, the higher is the standard deviation
which was exhibited by the saponins.
The total Antioxidant activity of the
haustorium obtained a standard deviation value
of 2.99 therefore the results of the experiments

4. Functional Components of the Coconut Haustorium
deviates only by 2.99 from the mean
(89.36±2.99) with a coefficient of variation (CV)
of 10%
Correlation Analysis is a statistical
technique used to determine the strength of the
relationship between two variables. X and Y. It
provides a measure of strength of the linear
relationship between two variables measured in
at least interval scale (Albacea, Reaño, Collado,
Comia, Tandang 2005). The dependent variables
are the functional components; Phenols,
Flavonoids and Saponins and the independent
variable was the total Antioxidant activity
(AOA) from the haustorium. Tannins and
alkaloids were not included in the correlation
analysis since both have negative results in the
experiment.
Computing for the correlation
coefficient (r) of phenols, the result absolute
value was 0.48 which means that there is a
moderate strength of linear relationship, basing
from the STAT 1 manual, UPLB (2005). The r2
gives the percentage of the total variation in the
values of the dependent variable that is
accounted for or explained by the independent
variable. From the table above r2 value of
phenols was 0.23 which means only 23% of the
total variation of the antioxidant activity is
explained by the amount of phenols.
For the Flavonoids, the computed
coefficient of correlation was r=0.68 which
means that there is a strong linear relationship
between the Flavonoids and AOA.
The computed coefficient of correlation
for saponins was r=0.87. The r2
value is equals
to 0.81, thus, 81% of the Antioxidant activity
was related to saponins which shows that there is
a very strong linear relationship between
Saponins and the AOA of the Haustorium.
SUMMARY AND CONCLUSION
The functional component of the
coconut haustorium; total phenols, Flavonoids,
saponins and the antioxidant activity were
determined except for tannins and alkaloids
which exhibited negative results due to the
absence of turbidity in the experiment.
In terms of the tests’ accuracy and
consistency the standard deviation was
computed. Phenols have a standard deviation of
17.34. Flavonoids with 18.43, and Saponins with
702.5. The saponins showed a huge dispersion or
deviation because of the high values of the data.
However, the total Antioxidant activity of the
Haustorium showed a small value of deviation
(2.99) and a coefficient of variation of 10%.
The functional component and the
antioxidant activity were correlated using the
coefficient of correlation (r). Phenols have a
moderate strength of linear relationship to
antioxidants (0.48). Flavonoids has a strong
linear relationship (0.68) and saponins has the
strongest linear relationship with the antioxidant
activity (0.87).
In conclusion, the antioxidant activity
(AOA) showed a low value of dispersion (Std
dev of 89.36±2.99) which can be attributed to the
accuracy of the experiment and saponins are
most likely related to it with the strongest linear
relationship.
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5. Functional Components of the Coconut Haustorium
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