Tropical Almond ( Terminalia catappa ) Leaf Extract as Effective Anti-

Hyperglycemic Activities in Rattus rattus (Rat)

A Concept Paper
Presented to
Dr. Fe M. Dela Cruz
College of Education
GRADUATE STUDIES
CARAGA STATE UNIVERSITY
Ampayon, Butuan City

In Partial Fulfillment
Of the Requirements of the Course
Science Investigatory Project

JULITO S. GULLE JR.

09451429768/09382223813
MSEd – Biology II

August 2019
 Chapter I
Introduction

Background of the Study

Tropical Almond Terminalia catappa is a fast-growing deciduous or semi-

evergreen tree, usually growing about 15 meters tall with specimens up to 40 meters
recorded. The canopy has a pagoda-like habit when young, but this becomes less
noticeable as the tree ages and the branches elongate and droop at the tips. The bole can
be straight or twisted, it is generally set with short woody spines, can be 100 - 150cm in
diameter and is often buttressed at the base with buttresses up to 3 meters high. An
important, multi-purpose tree, providing food, medicines and a host of other
commodities. This is one of the most common trees of many tropical coasts. It is widely
cultivated in the tropics, as a shade tree, for ornament and for its edible seeds. It is often
planted in avenues as a shade-tree, for which it is suitable because of its very regular
shape.

Plants grow best in the lowland tropics at elevations below 800 meters. It grows
best in areas where the mean maximum and minimum annual temperatures are within the
range 21 - 32°c, though it can tolerate 10 - 36°c. It prefers a mean annual rainfall in the
range 1,500 - 3,400mm, though can tolerate from 700 - 4,200mm. Succeeds in any
moderately fertile, well-drained soil in a sunny position. Prefers sands and loamy sands,
though it also does well on silts, loam, and even clays if the drainage is good. Prefers soil
that is neutral to moderately alkaline and rich in bases; however it will also grow in
strongly acid soils. A particularly useful plant for coastal gardens, being tolerant of salt
spray, established plants are drought tolerant. Prefers a pH in the range 5 - 7, but tolerates
4.3 – 8 (Tropical Plants Database, Ken Fern. tropical.theferns.info. 2019-08-28)..

Plants are fast growing, often in excess of 1 meter a year when young. Trees can
yield two crops a year in some areas, and in places have been known to exhibit more or
less continuous fruiting. A tree may yield around 5 kg of kernels per year. There has been
some selection in cultivating forms with larger seeds. The tree sheds its leaves all at once,
quite suddenly, usually twice a year (January or February and July or August). Unlike
most tropical trees, the leaves turn first yellow, then vivid red before falling, giving a
well-marked 'autumn color'. The inconspicuous flowers have a sweet, delicate fragrance.
Various species of biting and stinging ants have been found inhabiting hollow twigs on
this tree (Tropical Plants Database, Ken Fern. tropical.theferns.info. 2019-08-28).

Terminalia catappa L. belongs to the family Combretaceae. T. catappa is used

primarily as an ornamental shade, and salt-tolerant street tree, but the leaves provide food
for the Tasar silkworm, and the seeds are edible like almonds with sim Malay peninsular
and through the Canary islands this tree is known as the tropical almond. been claimed to
have therapeutic effects for liver related diseases. In India, it is used as cardiac stimulant.
Its leaves are widely used as a folk medicine in Southeast Asia for the treatment of
dermatitis and hepatitis. More and more pharmacological studies have reported that the
extract of T. catappa fruits have anticancer, antioxidant, antitranscriptase, anti-
inflammatory, antidiabetic effects and hepatoprotective activities components and related
mechanisms remain unknown. In the present work, phytochemical, antimicrobial and
invitro antioxidant activities were carried out from catappa bark methanol extract.
Antimicrobial activity of T. catappa bark extract was investigated against bacterial and
fungal cultures whereas inviro antioxidant activity was performed with DPPH method
(Praveena (2014).

Objectives

This research aimed to determine the phytochemicals content of Tropical Almond

(Terminalia catappa) leaves extract (TCL) and its effects as a cardiovascular stimulant of
rats/mice.

Hypothesis

HO. There is no significant effect of leaf extract as cardiovascular stimulant.

HO1. Leaf extract of Tropical Almond is effective for cardiovascular stimulant.

 Chapter II

Review of Literature

Glucose is derived from thediet and utilized by cells with the help of insulin.
Insulin insuffi-ciency elevates the blood glucose and causes hyperglycemia, whichleads
to diabetes. The ethanolic leaf extracts (300 mg/kg and500 mg/kg) decreased blood
glucose levels in diabetic rats. A similar effect has been found in treatment with different
extracts ofT.catappafruits (Nagappa et al., 2003), which confirm the anti-diabetic effects
ofT.catappa(Malviya et al., 2010). The secretion of insulin by beta cells of the pancreas
was affected by diabeticagents. On the other hand, the lower concentration of insulin
inthe toxin treated rat group was due to damage to the pancreatic cells, which were not
able to produce insulin due to DNA alkylation arising from the production of carbonium
ions (Wright et al.,1999). STZ-induced diabetes not only affects the pancreas, but
alsocauses damage to the liver, the kidneys and cardiac cells (Biswaset al., 2011). Related
studies have reported that the induction ofdiabetes in animals by beta cell destruction,
through the produc-tion of free radicals, leads to the alkylation of DNA and a conse-
quent increase in the blood glucose concentration (Takasu et al.,1991). In the present
study, there was a substantial increase inserum insulin levels of diabetic animals
afterT.catappa(300 mg/kg and 500 mg/kg) treatment. Hence, it might be recognized
thatsecondary metabolites of this plant extract stimulate the beta cells.

Ever since the dawn of civilization, medicinal plants have been of great
importance to the health of individuals, communities in Nigeria, and the whole world.
Man has continually investigated tropical and subtropical medicinal plants in order to
assess the importance of developing natural, sustainable, and affordable drugs and
cosmetics. The genus Terminalia L. are perennial shrubs or trees of the Combretaceae,
and nearly 200 species are identified. The genus is distributed in tropics and sub tropics
regions, a few species are found in Africa, northern Australia, Pakistan, India, Sri Lanka
and many other south Asian countries. Tropical almond (Terminalia catappa L.) is a
large tropical tree growing up to 35 m with an upright, symmetrical crown and horizontal
branches. This plant has been studied extensively. The bark is rich in tannins, the fruit is
bitter, acrid,astringent, an aphrodisiac. The leaves contain several flavonoids including
kaempferol and quercetin, saponins, and phytosterols. Fallen leaves of T. catappa have
been widely used in traditional medicine for the treatment of liver diseases, headache,
colic, and as a diuretic and cardiotonic T. catappa extracts have shown antidiabetic,
actinociceptive antiparasitic, antibacterial, and antioxidant activities, while the aqueous
extract of the bark is used traditionally to treat dysentery and diarrhea.

Traditional medicine has been practiced from many centuries, especially in India
tribal and rural areas people were used to treat several diseases, due to availability and
low cost and negligible side effects it occupies great importance in many formulations
and ayurvedic drugs. Nature has provided a source of medicinal agents for thousands of
years and an impressive number of modern drugs have been isolated from natural
sources, many based on their use in traditional medicine1 . There has been an increasing
the incidence of multiple resistances in human pathogenic microbial strains , largely due
to the indiscriminate use of synthetic antimicrobial drugs commonl treatment of
infectious diseases2 . The development of bacterial resistance to presently available
antibiotics has necessitated the search for new antibacterial agents, to overcome this
problem studies have been conducted with the various medicinal plants, screening
antimicrobial activity as well as for the discovery of new antimicrobial compounds3-4
and antioxidant activity.

Besides saponin, another group of phytochemicals, such astriterpenoid, quinon,

phenolic are also a common plant active compound that can be found in TCL and has
been reported to promote various activities like antistress, growth promotion, appetite
stimulation, tonic, and immune booster (Citarasu, 2010; Chakraborty et al., 2012;
Chakraborty et al., 2014). Meanwhile, tannins and flavonoid which were a presence in
TCL are the main group of plant phenolic compound that act as primary antioxidants or
free radical scavengers. According to Chansue and Assawawongkasem (2008), tannin
from T. catappa extract with water has the ability as an antibacterial substance for
ornamental fish. The tannin could inhibit the growth of bacteria in intestinal of fish by
binding iron and form a chelate. On the one hand, flavonoid is one of the most
widespread groups of phytochemicals in the plant that was shown to improve
reproduction of Japanese Medaka (Oryzias latipes) (Kiparissis, 2001). It acts as
antimicrobial properties on fish pathogen (Rattanachaikunsopon & Phumkhachorn,
2007), and as a growth promoter on juvenile Pargus major (Ji et al., 2007), Carassius
auratus (Ahilan. B et al., 2010), Catla catla (Kaleeswaran et al., 2011). Thus, the presence
of secondary metabolites in TCL might play a role in the survival and blood profile of
Betta sp.

Gao et al. identified various phytoconstituents from the fruits, seeds, and barks of
the T. catappa. The fruit has 1.95 g of protein, 12.03 g of carbohydrate, and 1.21 g of ash.
β-carotene (2,090 μg) and vitamin C (138.6 mg) are present in high amounts. The
mesocarp of fruits dehydrated by the sun having ash, protein, glucose, moisture, tannin,
carbohydrate, and oil with 3,434.5 kcal/kg calorific value is very essential for its nutritive
value. The seed is composed of fixed oil (51.2%), olein (54%), and stearin (46%). The
seeds yield 4.13% moisture, 4.94% crude fiber, 23.78% crude protein, 4.27% ash,
51.80% fat, and 16.02% carbohydrate; the total calorific value is 548.78 kcal. The bark
contains glycoside, cardiac tannins, volatile oils, saponin, steroid, glycosides, and
phenols. Classified in the oleic-linoleic acid group, the oils contain huge levels of
unsaturated fatty acids, exclusively oleic (up to 31.48%) and linoleic (up to 28.93%).
More recently, Mininel et al. isolated punicalagin (polyphenol), its derivatives, and other
several compounds in the leaves of T. catappa.

The leaves of T. catappa contain 1-degalloyl-eugeniin, 2,3-(4,4’,5,5’,6,6’-

hexahydroxy-diphenoyl)-glucose, chebulagic acid, gentisic acid, corilagin, geraniin,
granatin B, kaempferol, punicalagin, punicalin, quercetin, tercatain, tergallagin, terflavin
A, and terflavin B. The seeds contain carbohydrates, protein, fat, fiber, iron, ascorbic
acid, arachidic acid, β-carotene, linoleic acid, myristic acid, oleic acid, palmitic acid,
palmitoleic acid, stearic acid, phosphorus, potassium, niacin, riboflavin, thiamin, and
water. The fruit contains glucose, pentosans, corilagin, brevifolin carboxylic acid, β-
carotene, cyanidin-3-glucoside, ellagic acid, gallic acid, and tannin. Shikhamandloi et al.
identified quercetin in the leaf of T. catappa. The phytoconstituents like flavonoids,
carotenoids, and phenolic compounds may be responsible for the traditional use of this
plant.
 Chapter III

Methodology

The following method was adopted from the existing research articles. This
research is entitled The Effects of Terminalia catappa L. Leaves Extract on the Water
Quality Properties, Survival and Blood Profile of Ornamental fish (Betta sp) Cultured.

Plant Material

Terminalia catappa’s dried cut brown leaves were collected on the campus of
Carga State University main campus, Ampayon, Butuan City. In purpose of eliminating
the extraneous matter, the collected T. catappa’s leaves were washed with deionized
water and immediately undergo an air-dried process or dried in an oven at 40 degrees
Celsius for 12 hours. T. catappa’s powder (8 opening/cm passed) was obtained using a
mill. The powder was extracted with ethanol 95% for three days (100 g/L). After
filtration, the extract was evaporated by using rotary evaporator and stored at 4 degrees
Celsius until used as a crude extract.

Phytochemical Tests

Table 1. Animals and Experimental set up

Experimental Set-up

Control Group Experimental Group (Diabetic)

T. catappa L. extract T. catappa L. extract T. catappa L. extract No treatment of
10 mL. 20 mL. 30 mL. T. catappa L. extract
The preliminary phytochemical tests such as alkaloid, saponin, steroid,
triterpenoid, quinon, phenolic, tannin, and flavonoid were performed to detect the
presence of possible phytochemicals in the extract of T. catappa leaves.

Animals are divided into five groups. Group 1 is the control group, group 2, 3, 4
and 5 serve as the experimental group that administered with the same volume of
diabetic. Group 2 is treated with 10ml, group 3 is treated with 20mL, group 4 is treated
with 30mL, and group 5 receives no treatment of T. catappa L. extract. The rats are
monitored every 24 hours. Before and during the experiment rats were fed with standard
diet (Gold Mohr, Lipton India Ltd). After randomization to various groups and before
initiation of experiment, the rats were acclimatized for a period of 7 days under standard
environmental conditions of temperature, relative humidity, and dark/light cycle. Animals
described as fasting were deprived of food and water for 16 hours ad libitum Ahmed S.
M. et.,al 2005.

Statistical Analysis

The experiment was repeated three times with six animals for each group.
Statistical analysis was carried out by one way ANOVA using the standard statistical
software package of social science (SPSS) version 12.0. P0.05 was considered as the
level of significance Divya, N. et.,al (2018).

References
Ahilan. B, Nithiyapriyatharshini, A., & Ravaneshwaran, K. (2010). Influence of certain
herbal additives on the growth, survival and disease resistance of goldfish,
Carassius auratus (Linneaus) Tamilnadu. Journal of Veterinary and Animal
Sciences, 6(1), 5-11.

Chakraborty, S. B., Horn, P., & Hancz, C. (2014). Application of phytochemicals as

growth-promoters and endocrine modulators in fish culture. Reviews in
Aquaculture, 6(1), 1-19.

Chansue, N., & Assawawongkasem, N. (2008). The in vitro antibacterial activity and
ornamental fish toxicity of the water extract of Indian Almond leaves (Terminalia
catappa Linn.). KKU Veterinary Journal, 18(1), 36-45.

Citarasu, T. (2010). Herbal biomedicines: a new opportunity for aquaculture industry.

Aquaculture International, 18(3), 403-414.

Divya, N., Vijaya-Anand, A., 2014. Phytochemical investigation and in vitro antidiabetic
activity of Terminalia catappa leaves. Int. J. Phyto. Pharm. 4, 132–134. Edelman,
D.M.K., Olsen, T.K., Dudley, A.C., Harris, E.Z., Oddone, J., 2004. Utility of
hemoglobin A1c in predicting diabetes risk. J. Gen. Intern. Med. 12, 1175–1180.

Dr. Syed Mansoor Ahmed, Department of Pharmacology, M.M.U College of Pharmacy,

Karnataka, India. E-mail: mansoor276783@yahoo.com.

Dukes. Phytochemical and Ethnobotanical Database. 2008:11.

Gao J, Tang X, Dou H, Fan Y, Zhao X, Xu Q. Hepatoprotective activity of Terminalia

catappa L. leaves and its two triterpenoids. J Pharm Pharmacol. 2004;56:1449–55.

Hyttel, P., Sinowatz, F., & Vejlsted, M. (2010). Essentials of domestic animal
embryology. UK: Elsevier Health Sciences.

Ji, S. C., Takaoka, O., Jeong, G.-S., Lee, S.-W., Ishimaru, K., Seoka, M., & Takii, K.
(2007). Dietary medicinal herbs improve growth and some nonspecific immunity
of red sea bream Pagrus major. Fisheries Science, 73(1), 63-69.
Kaleeswaran, B., Ilavenil, S., & Ravikumar, S. (2011). Growth response, feed conversion
ratio and antiprotease activity of Cynodon dactylon (L.) mixed diet in Catla catla
(Ham.). Journal of Animal and Veterinary Advances, 10(4), 511-517.

Kiparissis, Y. (2001). Effects of flavonoids and other phytochemicals on fish Cypia

monooxygenases, embryonic and reproductive development, Trent University,
Canada.

Malviya, N., Jain, S., Malviya, S., 2010. Antidiabetic potential of medicinal plants.

Acta Pol.

Mandloi S, Mishra R, Varma R, Varughese B, Tripathi J. A study on phytochemical and

antifungal activity of leaf extracts of Terminalia cattapa. Int J Pharm Bio Sci.
2013;4:1385–93.

Meena, K., & Raja, T. K. (2006). Immobilization of Saccharomyces cerevisiae cells by

gel entrapment using various metal alginates. World Journal of Microbiology and
Biotechnolog, 22(6), 651-653.

Mininel FJ, Leonardo Junior CS, Espanha LG, Resende FA, Varanda EA, Leite CQ, et al.
Characterization and quantification of compounds in the hydroalcoholic extract of
the leaves from Terminalia catappa Linn. (Combretaceae) and their mutagenic
activity. Evid Based Complement Alternat Med 2014. 2014 676902.

Nagappa, A.N., Thakurdesai, P.A., Raob, N.V., Singh, J., 2003. Antidiabetic activity of

Terminalia catappa Linn fruits. J. Ethnopharmacol. 88, 45–50.

Praveena, K. (2014), Phytochemical, Anti-microbial and In-vitro Antioxidant activity of

Terminalia catappa, International Journal of Pharmacy & Life Sciences, 5(2), pp.
3325-3329.

Rattanachaikunsopon, P., & Phumkhachorn, P. (2007). Bacteriostatic effect of flavonoids

isolated from leaves of Psidium guajava on fish pathogens. Fitoterapia, 78(6),
434-436
Tropical Plants Database, Ken Fern. tropical.theferns.info. 2019-08-28.
<tropical.theferns.info/viewtropical.php?id=Terminalia+catappa

Vučurović, V. M., & Razmovski, R. N. (2012). Sugar beet pulp as support for
Saccharomyces cerivisiae immobilization in bioethanol production. Industrial
Crops and Products, 39, 128-134.