Talisay Almond (Terminalia catappa L.

) As pH Indicator

ABSTRACT
A convenient method of extraction of anthocyanin from Talisay almond (terminalia
catappa L.) red leaves extract has been developed using aqueous solution. It has been shown
that by changing the pH of the extract, which the hue color changes from dark red to red
orange. In the present study we have made use of anthocyanin extracted from talisay red
leaves as natural indicator for acid-base indicator. Significant changes were observed and
usefulness of talisay red leaves as natural acid-base indicator has been exemplified.

Chapter 1

Introduction

All pH indicators change color depending on whether they donate or accept proton
(acids are proton donors and bases are proton acceptors). A pH indicator is just a weak acid
with differently colored acid and conjugate base forms. Therefore pH indicators are themselves
acids or bases. Many flowers, fruits, and vegetables contain chemical substances that change
color in solutions of different pH. The plant pigments can act as acid-base indicators.
The blue and red pigments of flowers were isolated and extensively studied by Willsatter
in 1913. Many indicators have been extracted from plants since then. Selective natural
indicators (such as litmus) to indicate specific pH levels have been developed based on their
changes. The substances in the plant produces such as tea, red cabbage or grapes react with
acids or bases resulting in the changes at the molecular level which causes their color to be
different at different pH. Red cabbage juice has been used by I. Jansons, (2001) as a natural pH
indicator which is known to contain anthocyanin’s which has pigments that react in a different
way to acids and bases.
Apples, autumn leaves, strawberries and cranberry juice appear red and blueberries,
cornflowers and violets are appear blue while grapes, blackberries and red cabbage appear
purple due to the presence of cyaniding based colorant molecules.
Anthocyanin is probably the most readily available acid/base indicator, it is the plant
pigment that makes red cabbage pucornflowers, blue, and poppies red. It changes color from
red in acid solution to purplish to green in mildly alkaline solution to yellow in very alkaline
solution. Researchers who have used anthocyanin as acid-base indicators for economic use of
the pigments of some common flowers as acid/alkali indicators Bose, B (1983), have been
shown by using common flowers like Hibiscus rosa sinensis, Ipomoea fistulosa and Clitoria
turnetea contain various anthocyanin pigments in the form of purple-red, violet and blue colors.
The acetone extracts of the pigments turn red in presence of acid and show absorption max. at
500 nm (H. rosa sinensis) r at 525 nm(I. fistulosa and C. turtenetea). The extracts when treated
with alkali turn green and show a greater absorbance in the range 575-625 nm depending upon
the flowers used.
Indicators are pigments or dyes that can be isolated from a variety of sources including
plants, fungi, and algae.(4,5) Virtually any plants that is red, blue, or purple in color contains a
class of organic pigments known as anthocyanin that can change color with pH (6). Some
naturally colored substances change colors when acidity or alkalinity of their environment
changes, for example, grape juice, brown tea, and some flower pigments. These substances
are called acid or base indicators.(7) Indicators change color at a particular stage of chemical
reaction.(8) A number of commonly used indicators in the laboratories are methyl red, methyl
range, phenolphthalein, phenol red, methyl yellow, pentamethoxy red, bromophenol blue,
thymol blue and so forth. (9)Most type indicators are available for different types of titrimetric
analyses. For acid-base titrations, organic dyes which are either weak acids or bases, serve
excellently as indicators. (1, 3)

Acid-base indicators are substances that are revealed through the characteristic color
which changes the degree of acidity or alkalinity of solutions. (10, 11) The choice of an indicator
for a particular titration depends on the characteristic of the neutralization curve. Majority of
indicators in use today are synthetic. A synthetic indicator in man-made chemical substance in
the laboratory which is used to determine pH of a substance such as litmus paper. (12, 13)
litmus paper contains naturally occurring substances that indicate pH levels but the item as a
whole is made in the laboratory such as methyl red, methyl orange, penolpththalein, phenol red,
methyl yellow, pentamethoxy red, bromophenol blue, thymol blue. Synthetic indicators have
certain disadvantages such as high cost, availability and chemical pollution, hence natural
indicators obtained from various plant parts like flowers, fruits and leaves will be more
advantageous. (14, 15) In addition, some of these synthetic indicators have toxic effects on
users such as diarrhea, pulmonary edema, hypoglycema and pancreatic and they can result in
abdominal cramps, skin rash, eruptions, erythema, and epidermal necrosis and cause
environmental pollution. (16, 17)
 On the bases of these rationales of the hazardous effects of synthetic indicators, there
has been an increasing interest in the search for alternative sources of indicators from natural
sources of plant origin and probability cheaper, readily available, easy to extract, less toxic to
users and environmentally friendly. Quite a number of dyes are obtainable from natural
products. For instance, in Nigeria, several researchers have extracted different types of dyes
from a variety of local plants. Also, several studies by various investigators have reported the
effectiveness of natural indicators in acid-base titrations. Some flowers such as Rose,
Allamanda and Hibiscus work in nature like litmus paper, changing color in the presence of
acids or bases, these flowers are usually mildly aciic or alkaline themselves and they change
color when mixed with a substance that has an opposite pH. (23). others are plants and leaves
such as red cabbage extract, blueberry juice, black tea, beet juice and tomato leaves.
An indicator does not change color from pure acidic to pure alkaline at specific hydrogen
ion concentration, but rather color change occurs over a range of hydrogen ion concentrations.
This is the color change interval expressed as the pH range. A natural indicator is a natural
substance usually from plant origin that can be used to determine the pH of another substance
(6,21). Hence in this study, the researcher aim to evaluate the properties of talisay almond,
terminalia catappa L. leaves in order to ascertain their analytical potentials as indicators.

Keywords: talisay, pH indicator, anthocyanin

 Chapter 2

Methodology

The acidic or basic nature of materials is usually determined using pH meter and
commercially-sold or synthetic indicating means. In this study, an eco-friendly pH indicator was
prepared from the aqueous extract of talisay almond (Terminalia catappa L.) red leaves
containing anthocyanin pigment. The talisay pH indicator paper was calibrated using buffer
solutions pH 0 to 14 to come up with a colored chart. This served as reference in the
subsequent effectiveness and acceptability tests conducted of the product. The indicator
produced showed a highly acceptable response from the respondents proving that it can be a
good alternative to commercially-sold pH indicating means. The shelf-life test also revealed that
it could withstand the storage time of 3 months without losing its effectivity. These findings have
significant implications to further refine the product and prolong its storage time, and
consequently, the production of talisay pH indicator paper.

Experimental Component
Preparation and storing. The indicator was prepared by collecting and weighing about
100 grams of fresh and clean talisay red leaves; grinding the sample in mortar and pestle and
then mixing the mush with 50 mL water in an amber-colored bottle. The solution mix for 7 days.
The mixture was then filtered.
Technically, the acidity or alkalinity of solutions is determined through its pH. The “power
of hydrogen” or simply pH uses the mathematical logarithm of the concentration of hydrogen
ions in solutions (Brown, Le May, Bursten, & Murphy, 2009). This chemical measurement
makes use of various indicating ways ranging from the sophisticated bench tester digital, pH
meter, and hydron paper which can be used in school laboratory activities, in analyzing
commonly used household materials, in farming, and in small scale industries. The most
popular and common litmus and pH hydron paper establishes as the acidic or basic nature of
substances and solutions by causing a chemical reaction that produces visual specific color
change to the chemical in the paper thus, “indicating” the nature of the material being tested
(Brady & Holum, 1988).
Sold in the market are several types of expensive commercial indicator such as methyl
orange that can distinguish with multiple color changes. These indicator have been prepared by
infusion in colored extracts from plant parts. For hundreds of years these isolated natural
indicators had been utilized even before chemists manufactured acid-base indicators. In fact,
Robert Boyle first reported the use of natural dyes as acid-base indicators in 1664 in his
collection of essays Experimental History of Colors (Bhise et al, 2014).
The prominent colors in plant extracts are due to the presence of naturally occurring
organic substances such as anthocyanins, azo compounds, flaovenes, flavonol, xanthine,
among others. Literature reveals that extracts containing anthocyanin are pH sensitive in that
they exhibit different color reactions when added to an acid or base (Khan & Farooqui, 2011). In
the Philippines there are plenty of plants that can be sources for preparing indicators. One of
these is talisay almond, terminalia catappa Linn. of the Family Crombetaceae and locally called
talisoy. This plant does not seem to have varieties. However, it has been reported to have a
sweet nature occurring in the cultivated and semi cultivated state throughout the country
(Quisumbing, 1978). This tropical attractive, long-lived tree reaches 5-10 meters in height and is
abundant in Luzon. It has a large trunk which divides into a number of upright branches (Padua,
1996). Its flowers are 10-22 mm long, borne in small hairy panicles emerging directly from the
trunk and oldest, thickest branches and on some twigs, as do the cluster of its fruits (World Agro
Forestry, 2015). They are small, fragrant, auxiliary or cauliflorous, 5-petalled, yellowish-green
color or red color, the source of pigments that contain chemicals from the naturally-colored
anthocyanin family of compounds (Ecocrop FAO, 2007). Anthocyanins are a large group of
naturally colored red-blue family of flavonoids found in pigments, responsible for the attractive
colors of plant parts. They constitute the largest group of pigments in nature. They are the
universal water soluble red pigments of flowers, fruits and leaves (Sharp, 1987). Chemically,
they are glycosides and their aglycones, the sugar free pigments, are identified as the
anthocyanidins (Khan & Farooqui, 2011). The known anthocyanin isolated from talisay almond,
terminalia catappa is the cyanidin-3-O-beta-D-glucoside (as cited in Gunasegaran, 1992).This
anthocyanin cation is a cyanidin cation linked to a ß-D- glucosyl moiety at position 3 (Chemical
Entities of Biological Interest [ChEBI], 2014)
From these evidences and owing to its abundance, it is viable and beneficial to prepare
an eco-friendly and affordable talisay pH indicator paper as a substitute for expensive pH
indicating means. Such will be used as an instructional material for household or domestic
activities and local industries. Explicitly, this study aimed to (1) prepare pH indicator from talisay
almond (terminalia catappa L.) red leaves extract; (2) establish the effectiveness of the
indicator.
 The present work is both experimental and descriptive. The experimental component
involved the following steps: preparing, storing, calibrating and testing for effectiveness of
talisay pH indicator. The subsequent descriptive part is to gather data which were encoded,
analyzed and interpreted.
Testing for effectiveness. An experiment was performed using talisay pH indicator and
its color chart for reference to screen its effectiveness and acceptability compared to the
commercially methyl orange. This practical activity has shown and identified the nature of some
commonly used chemicals in the laboratory.
Three spot plate were arranged horizontally in three sets: A, B, and C on the working
table at the Bio Laboratory. In set A, the first spot in plate contained talisay pH indicator; the
second had methyl orange indicator. The color of talisay indicator were recorded when they
were tested on acid solution. For first spot plate test, 0.1 M Hydrochloric acid, HCl, 0.1 M H 2SO4
and acetic acid was added. All results obtained were recorded and tabulated.
The same procedure of indicator distribution was done in Sets B. The initial colors of the
different indicators were recorded when they were tested on basic solution. For first spot plate
test, Naoh, and Nh4oh was added. All results obtained were recorded and tabulated.
The same procedure of indicator distribution was done in Sets C. The initial colors of the
different indicators were recorded when they were tested on salt solution. For first spot plate
test, 0.32N, 0.8 N, 0.16N sodium thiosulfate was added. All results obtained were recorded and
tabulated.
 Results

In the activity, the students determined the indicator’s usefulness as an instructional

material and as substitute in identifying the nature of some commonly used materials in the
laboratory. They evaluate the product and give their feedback. These feedbacks were gathered,
tabulated, analyzed and interpreted as follows:
Interpretation
Acid talisay indicator is highly acceptable
Base talisay indicator is acceptable
Salt talisay indicator is slightly acceptable
- talisay indicator is not acceptable in methyl orange indicator thus may acceptable in
phenolphthalein indicator

Discussions

Aqueous extract of tropical almond (terminalia catappa L.) red leaves containing
anthocyanin was prepared and used to infuse strips of filter paper to be utilized as pH indicator.

Calibration of the Indicator Paper

The standardization of the locally prepared indicator paper showed a clear picture
presentation of its different color reactions with buffer solutions from pH 0 to 14. Figure 2
illustrates the color chart produced for the indicator. This representation consequently served as
reference for the succeeding investigations on the effectiveness of the indicator paper.
Figure 2. Talisay pH Indicator Color Chart

Establishing Effectiveness of Talisay pH Indicator Paper

Table 1 showed the effectiveness of the indicator in determining the acidity or alkalinity
of some substances as compared with other indicating means. The reaction of talisay pH
indicator on the test materials was specific and comparable. Litmus paper, on the other hand,
gave a descriptive result, that is, the test material is either acidic or basic.

Table 1
pH of Some Common Laboratory Chemicals

Materials Talisay Indicator Paper Litmus Paper

(base on the color chart)

ACIDS :
HCl dark red 4(red) red remained red

H2so4 red 7(dark red) light red remained red

Acetic acid dark red 5(dark red) red remained red

Base:
Naoh dark blue 14(light blue) remained blue blue

Nh4oh lighter shade 6 (slightly No change in No change in

of blue basic) color color
Salt:
0.32N light red violet No change in No change in
Light violet color color

0.8 N light orange No change in No change in

Light orange color color

0.16 yellow Remained Remained

Orange yellow yellow yellow
 Recommendations

The indicator can stay for a period of time, 3 months in the case of the present study.
The storing did not affect its quality and affectivity. Hence, it is recommended that the
experimental component of this study be continued. The focus of which should be in the
refinement of the color chart as reference for future analysis. Along this line, there is a need to
investigate further the best proportion of the plant’s leaves extract to the extracting solvent. And
also, it is recommended to test for shelf-life in one year to determine talisay pH indicator storage
capacity. These steps would surely ensure the quality and usability of the product.
 References

Padua, L.S (1996). Philippine plants: Medicinal Plants. Manila, Philippines. Pundasyon sa
Pagpapaunlad ng Kaalaman sa Pagtuturong Agham, Ink (Foundation for the
Advancement of Science Education, Inc.).
Quisumbing, E. (1978). Medicinal plants of the Philippines. Caloocan City: Katha Publishing C.,
Inc.