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Journal of Pharmacognosy and Phytochemistry 2015; 4(2): 40-43
E-ISSN: 2278-4136
P-ISSN: 2349-8234
JPP 2015; 4(2): 40-43
Received: 20-04-2015
Accepted: 24-05-2015
Geedhu Daniel
PhD Scholar, Department of
Biochemistry, Kongunadu Arts
and Science College, Coimbatore-
641029, Tamilnadu, India.
S. Krishnakumari
Associate Professor in
Biochemistry, Department of
Biochemistry Kongunadu Arts
and Science College, Coimbatore-
641029, Tamilnadu, India.
Correspondence:
S. Krishnakumari
Associate Professor in
Biochemistry, Department of
Biochemistry Kongunadu Arts
and Science College, Coimbatore-
641029, Tamilnadu, India.
Quantitative analysis of selected primary metabolites in
aqueous hot extract of Eugenia uniflora (L.) leaves
Geedhu Daniel, S. Krishnakumari
Abstract
Purpose of present study was to evaluate the presence of selected primary metabolites in aqueous hot
extract of Eugenia uniflora leaves. Primary metabolites like lipids and amino acids were estimated using
standard procedures. Quantitative analysis is very essential for identifying the compounds present in the
medicinal plants. The results obtained from the present study provides evidence that aqueous hot extract
of Eugenia uniflora leaves contains various primary metabolites and this justifies the use of this plant
species as traditional medicine for treatment of various diseases. The results are very much encouraging
but scientific validation is necessary before being put into practice.
Keywords: Primary metabolites, Lipids, Amino acids, Aqueous extract, Eugenia uniflora.
1. Introduction
Plants have been an integral part of traditional medicine across the continents since time
immemorial. Medicinal plants have their values in the substances present in various plant
tissues with specific physiological action in human body. Many of the plant species that
provide medicinal herbs have been scientifically evaluated for their possible medicinal
applications. India is endowed with a rich wealth of medicinal plants. India recognizes more
than 2500 plant species which have medicinal values
[1]
. Plants are like natural laboratories
where a great number of chemicals are biosynthesized and in fact they may be considered the
most important source of chemical compounds.
The identification of plants is useful to human beings from natural strands commenced in
prehistoric studies. Experiments and trails are the two main ways through which humans have
learnt various uses of the plants
[2]
. In recent times, focus on plant research has increased all
over the world and a large body of evidence has collected to show immense potential of
medicinal plants used in various traditional systems. More than 13,000 plants have been
studied during the last 5 year period
[3]
. Over three-quarters of the world population relies
mainly on plants and plant extracts for health care.
According to the Food and Agriculture Organization (FAO), more than 50,000 plant species
are used in the traditional folk medicine throughout the world
[4]
. The drugs are derived from
the whole plant or from different parts like leaves, stem, bark, root, flower, tuber and seed etc.
More than 30% of the entire plant species, at one time or other was used for medicinal
purposes. It has been estimated that in developed countries such as United States, plant drugs
constitute as much as 25% of the total drugs, while in fast developing country such as India,
the contribution is as much as 80%
[5]
.
Since ancient times, people have been exploring the nature particularly medicinal plants in
search of new drugs. Medicinal plants are used by 80% of the world population for their basic
health needs. India is the birth place of renewed system of indigenous medicines such as
Siddha, Ayurveda and Unani. Traditional systems of medicines are prepared from a single
plant or combinations of more than one plant. This efficacy depends upon the current
knowledge about taxonomic features of plant species, plant parts and biological property of
medicinal plants which in turn depends upon the occurrence of primary and secondary
metabolites
[7]
.
Plant synthesizes a wide range of chemical compounds which are classified based on their
chemical class, biosynthetic origin and functional groups into primary and secondary
metabolites. Primary metabolites directly involved in growth and development while
secondary metabolites are not involved directly and they have been worked as biocatalysts.
Primary metabolites are widely distributed in nature, occurring in one form or another in
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Journal of Pharmacognosy and Phytochemistry
virtually all organisms. They are like chlorophyll, amino acids,
nucleotides, carbohydrates etc., which have a key role in
metabolic processes such as photosynthesis, respiration and
nutrient assimilation. They are used as industrial raw materials
and food additives.
Eugenia uniflora L. is a widely distributed tree in South
American countries, mainly in Brazil, Argentina, Uruguay, and
Paraguay
[8]
. Its leaves are used in popular medicine as
infusion in the treatment of fever, rheumatism, stomach
diseases, disorders of the digestive tract, hypertension, yellow
fever, and gout. It may also reduce weight, blood pressure, and
serve as a diuretic
[9]
. Pitanga fruits, also known as Brazilian
cherry or Suriname cherry, contain various volatile compounds
that are also found in the essential oil of pitanga leaves
[10, 11]
.
Like the leaves, pitanga fruits may also have health benefits. In
the Brazilian food industry, pitanga fruits have mostly been
used to produce juice and frozen pulp. Pulp production has
high economic potential because the product has consumer
appeal and high concentrations of antioxidant compounds,
such as anthocyanins, flavonols, and carotenoids
[12]
.
Plants produce the majority of the world's lipids, and most
animals, including humans, depend on these lipids as a major
source of calories and essential fatty acids. Like other
eukaryotes, plants require lipids for membrane biogenesis, as
signal molecules, and as a form of stored carbon and energy.
In addition, soft tissues and bark each have distinctive
protective lipids that help prevent desiccation and infection.
Plant lipids also have a substantial impact on the world
economy and human nutrition. More than three-quarters of the
edible and industrial oils marketed annually are derived from
seed and fruit triacylglycerols. These figures are particularly
impressive given that, on a whole organism basis, plants store
more carbon as carbohydrate than as lipid. Since plants are not
mobile, and since photosynthesis provides fixed carbon on a
regular basis, plant requirements for storage lipid as an
efficient, light weight energy reserve are less acute than those
of animals.
The amino acids have several roles in plants, for example they
act as osmolytes, detoxify heavy metals, regulate ion transport,
stomatal opening, affect synthesis and activity of enzymes,
gene expression and redox homeostasis
[13]
. Positively charged
polyamines are involved in the stress response through their
interaction with the negatively charged macromolecules, such
as DNA, RNA, proteins and phospholipids, resulting changes
in the physical and chemical properties of the membranes, in
the structure of nucleic acids and in the enzyme activities
[14]
.
In addition, polyamines are able to detoxify the reactive
oxygen species accumulating during abiotic stress.
Considering the potential pharmacological benefits of Eugenia
uniflora, the aim of the study was to quantitatively estimate
primary metabolites like lipids and amino acids in aqueous hot
extract of Eugenia uniflora leaves.
Materials and Methods
Plant material
Fresh leaves of Eugenia uniflora (Linn), Family- Myrtaceae,
were collected from Wayanad district, Kerala during the
month of April 2014. Taxonomic authentication was done by
Dr. V.S Ramachandran, Taxonomist, Department of Botany,
Bharathiar University, Coimbatore, Tamil nadu, India.
Sample Processing
The leaves were washed, shade dried at room temperature and
powered in a mixer grinder.
Hot Water Decoction: 10g of the powdered sample was
dissolved in 100ml of distilled water which was boiled for one
and half hours and filtered. The decoction was stored at 4 °C
for further usage.
Quantitative estimation of Lipids
Lipids are an essential constituent of all plant cells. The
vegetative cells of plants contain 5 to 10% lipid by dry weight,
and almost all of this weight is found in the membranes.
Membrane lipids are important for improvement of
photosynthesis against high temperature stress and improved
photosynthesis means improved stress tolerance as well
[14]
. In
the present study free fatty acids, total cholesterol,
phospholipids, triglycerides were estimated using standard
procedures.
Table 1: Qunatitative estimation of Lipids
Parameters
References
Free fatty acids
Horn and Mehanan,1981
[15]
Total cholesterol
Parekh and Jung,1970
[16]
Phospholipids
Rouser,1970
[17]
Triglycerides
Rice,1970
[18]
Quantitative estimation of Aminoacids
Amio acids have traditionally been considered as precursor
and constituents of proteins. Many amino acids also acts as
precursor of other nitrogen containing compound eg:-nucleic
acids
[19]
. Amino acids such as tryptophan, methionine,
histidine, proline and arginine were quantitatively estimated
using standard procedures.
Table 2: Quantitative estimation of Amino acids
Parameter
References
Tryptophan Methionine Proline
Sadasivam and Manickam, 1996
[20]
Histidine
Kapeller and Adler,1933
[21]
Arginine
Sakaguchi,1925
[22]
Statistical Analysis
All the analyses were performed in triplicate and the results
were statistically analyzed and expressed as mean (n=3) ±
standard deviation (SD).
Results
Lipids are the major form of carbon storage in the seeds of
many plant species. Lipids are the most effective source of
storage energy, function as insulators of delicate internal
organs and hormones and play an important role as the
structural constituents of most of the cellular membranes.
Free amino acids and polyamines take part in several
metabolic processes and they are involved in the protection
against abiotic stresses.
The results obtained from the present study are shown in Table
3 and Table 4.
Table 3: Quantitative estimation of lipids
Primary
metabolites
Aqueous hot extract of Eugenia uniflora
leaves(mg/g)
Triglycerides
1.88 ± 0.08
Total cholesterol
0.67 ± 0.07
Free fatty acids
2.70 ± 0.10
Phospholipids
0.93 ± 0.07
Values are expressed by mean ± SD of 3 Samples
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Journal of Pharmacognosy and Phytochemistry
Table 4: Quantitative estimation of Amino acids
Primary
metabolites
Aqueous hot extract of Egenia uniflora
leaves(mg/g)
Tryptophan
0.70 ± 0.04
Methionine
0.19 ± 0.04
Histidine
1.06 ± 0.05
Proline
0.38 ± 0.04
Arginine
1.84 ± 0.08
Values are expressed by mean ± SD of 3 Samples
Discussion
In the present study hot water extract of Eugenia uniflora
shows higher amount of free fatty acids (2.70 ± 0.10 mg/ g)
followed by triglycerides (1.88 ± 0.08 mg/g).The fatty acid is
found in every cell of the plant and is essential to growth. As
part of complex lipids, fatty acids are also important for
thermal and electrical insulation, and for mechanical
protection. Triglycerides make up the structure of all vegetable
oils and fats found in nature. It acts as energy reserve when
stored as adipose tissue also acts as insulator, shock protection.
Phospholipids are important components of the lipid bi-layer
of the cell membrane of all cells. The cell membrane has an
essential general role of maintaining cell order and integrity
and a number of disease control mechanisms involve
compounds that directly (by partitioning into the membrane
and inducing disorder) or indirectly (by inhibiting fatty acid
biosynthetic pathways) target the phospholipids of the cell
membrane
[23]
. The level of Phospholipids in the aqueous hot
extract of Eugenia uniflora leaves was 0.13 ± 0.07mg/g. The
level of cholesterol was very low compared to others 0.67 ±
0.07 mg /g.
Tryptophan is an essential amino acid which acts as building
blocks in protein biosynthesis. In addition tryptophan
functions as a biochemical precursor for many compounds
[24]
.
The level of tryptophan in aqueous hot extract of Eugenia
uniflora leaves was 0.70 ± 0.04mg/g.
Methionine is needed to produce two sulphur containing
amino acids cysteine and taurine which helps the body to
eliminate toxins, build up strong, healthy tissue and promote
cardiovascular health
[25]
. The level of methionine in the
aqueous hot extract of Eugenia uniflora leaves was 0.19 ± 0.04
mg / g.
Histidine is found abundantly in haemoglobin.It has been used
in the treatment of reheumatoid arthritis, allergic diseases,
ulcers and anemia. Deficiency can cause poor hearing
[26]
. The
level of histidine in the aqueous hot extract of Eugenia
uniflora leaves was 1.06 ± 0.05 mg/g.
Proline is a proreogenic aminoacid with an exceptional
conformational rigidity and is essential for primary
metabolism
[27]
. Proline accumulation has been reported during
conditions of drought
[28]
high salinity
[29]
high light and UV
irradiation
[30]
, heavy metals
[31]
, oxidative stress
[32]
and in
response to biotic stresses
[33, 34]
. The level of proline in
aqueous hot extract of Eugenia uniflora leaves was 0.38 ±
0.04mg/g.
Arginine play an important role in the healing of wounds, in
muscle growth and in fetal and child development. It turns into
nitric oxide in the body and cause vasodilation, a relaxant of
arterial walls and facilitiates blood flow. The level of arginine
was more compared to other amino acids in the aqueous hot
extract of Eugenia uniflora leaves was 1.84 ± 0.08 mg / g.
The values of amino acid concentration in this study showed
that aqueous hot extract of Eugenia uniflora leaves contains
considerable amounts of lipids and amino acids and confirmed
that the nutritional quality of the sample was commendable
and may included in treating various diseases.
Conclusion
Plants and its products are used as medicine from the ancient
time1. Recently there has been a shift in universal trend from
synthetic to herbal medicine
[35]
. It is estimated by the World
Health Organization that approximately 75-80% of the world's
population uses plant medicines either partly or entirely as
medicine. Interest in plant derived drug increases mainly due
to the increasing use, and misuse, of existing synthetic drugs.
This poses the need for search and development of new drugs
to cure diseases. The chemical substances of the medicinal
plants which have the capacity of exerting a physiologic action
on the human body are the primary features. The bioactive
compounds of plants compounds are considered to be most
important. The phytochemical research that has been done
based on the ethno-pharmacological information forms the
effective approach in the discovery of new medicinal agents
from higher plants.
The results obtained in the present study indicate Eugenia
uniflora leaves have the potential to act as a source of useful
drugs because of presence of various phytochemical
components such as various lipids and amino acids. The
results are very much encouraging but scientific validation is
necessary before being put into practice.
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