Investigation
for Bioactive Compounds of
Berberis Lyceum Royle �and Justicia Adhatoda L.
������������������������������������������
M. Gulfraz, M. Arshad,
N. Nayyer, N. Kanwal and U.
Nisar
ABSTRACT
Gulfraz, M. and M. Arshad (Associate
Professors, Department of Biochemistry, University of Arid Agriculture,
Key Words: Berberis
lyceum, Justicia adhatoda,
Chemical analysis, medicine.
INTRODUCTION:
��������� �Medicinal plants are plants whose extracts can
be used directly or indirectly for the treatment of different ailments.
Therefore, the use of traditional medicine and medicinal plants in most
developing countries, as a basis for the maintenance of good health, has been
widely observed (Edward,2001). Scientists throughout
the world are trying to explore the precious assets of medicinal plants to help
the suffering humanity. Furthermore, in the world more than 30% of the
pharmaceutical preparations are based on plants (Shinwari
and Khan, 1998).
���������� However, an increasing
reliance on the use of medicinal plants in the industrialized societies has
been traced to the extraction and development of several drugs and
chemotherapeutics from these plants.������������������������������������������������������������������������������������������
The use of medicines from plants in the form of local medicine dates
back to 4000-5000 B.C. While the medicinal values of these plants are due to
the presence of small doses of active compounds which produces physiological
actions in the human and animal body (Zaidi, 1998).
Some of the important bioactive compounds found in medicinal plants are
alkaloids, glycosides, resins, gums, mucilages etc.
(Sack and Forehlich, 1982). It was observed that
developed countries mostly imports raw materials of valuable medicinal plants
from developing countries. Where they are screened, analyzed and used in drug
preparations, and returned as high priced medicines to developing countries
(Shinawie,2002).
������������ In
����������� Berberis
lyceum is locally
known as simbuli or simbulu
belonging to family Berberidaceae. It is about 4-6 feet
in height with thorny branches. The leaves are somewhat obviate, with ciliated
teeth on their margins. The flowers are drooping racemes, with yellow petals.
The berries (fruit) grow in loose bunches (Zaidi,1998).
���������� Berberis
lyceum is valued mainly for its fruits and roots, which contain alkaloids
like berberine and plamitine.
These alkaloids are effective against eye diseases, febrifuge, and piles (Gosh et
al,1990). Whereas,an extract made from its roots (known as �rasaunt�) which is being used against many infections
including eye�s disorders. (Chopra et al. 1998).
Whereas in some areas of
��������� Justicia adhatoda is one of the most important
specie and dominant�
vegetation of hilly areas of
������� Chemical compounds found in leaves and
roots of this plant includes essential�
oils,� fats,� resins, sugar, gum, amino acids, proteins and
vitamin�C`etc. (�
Dymock,1972).
������� The medicinal properties of
Adhatoda vasica
Nees are well known in
��������� �The leaves are mostly used in the treatment of
respiratory disorders in Ayurveda.The alkaloids, vasicine and vasicinone present
in the leaves, possess respiratory stimulant activity(Baquar,1997).
Whereas, vasicine, at low
concentrations, induced bronchodilation and
relaxation of the tracheal muscle. However,� at high concentrations, vasicine offered significant protection against
histamine-induced bronchospasm in guinea pigs. Vasicinone, the auto-oxidation product of vasicine has been reported to cause bronchodilatory
effects both in vitro and in vivo (Shinawie,
2002).
� �������Therefore keeping in view the importance
of these valuable medicinal plants, the present study was undertaken with the
following aims and objectives.
1. To assess the bioactive compounds
of Berberis lyceum royle
and Justicia adhatoda.
2. To compare the chemical analysis of compounds found
in Berberis lyceum fruits and Justicia adhatoda.
3. To highlight the values of these
bioactive compounds against certain infections in human population.
MATERIALS AND METHODS
Collection of samples
�����������
�
�����������In order to analyze the
bioactive compounds present in root and fruit of Berberis
lyceum and leaves, fruit
and root of Justicia adhatoda,the
plant samples were collected from different localities of Murree, and surrounding hilly areas. Whereas chemical
analysis of the root, leaves and fruit samples were
carried out with following procedures.
Preparation of samples
���������� After collection of the roots , leaves and fruits samples of Berberis lyceum and Justicia adhatoda were washed
and subjected to sun dried, followed by oven dried. Finally the samples were
crushed and� converted
into powdered form and saved for further analysis.
Chemical analysis of root, leaves and fruit samples
����������� Both roots and fruits samples were
analyzed for protein, carbohydrate, lipid, vitamins, and fibers. Sodium, Potassium, Calcium, phosphorus.Sulphur,
Iron and Zinc by routine
chemical analysis. Whereas alkaloids of these valuable plants species
were separated by chemical extraction methods followed by Column and thin layer
chromatography.
Instrumentation
��������� In order to extract and purifying
alkaloids from roots, leaves and fruits samples, following procedure were
adopted.
���������� Fifty grams (each of roots, fruits
and leaves) samples were soaked in the ethanol (80%) for 24 hours and filtered.
The ethanol was evaporated and half volume of NaOH
(3-4%) was added. The pH of the mixture was adjusted to 10 with NaOH. The mixture was run through a column using silica gel
to separate the alkaloids through column chromatography,
those were further identified on thin layer chromatography using reference
standards.
��������� The concentration level of these
alkaloids was determined with the help of spectrophotometer at 650 nm� and that was
compared with standard alkaloid compounds. Finally the PH of alkaloids were
obtained and were compared with standard alkaloids .
(Source: Vennestrom and Klayman,1988)
Vasicine
Vasicinone
(Source: Atal,1980)
Structures of alkaloids
RESULTS AND DISSCUSSION:
�������� Results
of biochemical compounds found in roots, leaves and fruits of Berberis lyceum and Justicia
adhatoda is given in tables 1-5 whereas
comparison of these Bio active compounds are given in Figure 1 and 2.
�������� Higher concentration of alkaloids was
found in roots as compared to the fruits (Tables 1 and 2). Furthermore, high
concentration of proteins (4.5 % ), fat (2.6 %), fiber
(2.5%) ,sodium (1.5% ) calcium (2.2%), sulphur
(0.2%), iron (0.3% zinc (0.3%), palmitine (3.1%) and berberine (4.5% were present in
the roots. However, the level of these chemicals was low in fruits except sugar
(4.5 %), and vitamin C (0.8%) was high in fruits as compared to roots (Tables 1
and 2). Furthermore, the comparison of these compounds are
presented in figure 1.
��������� The pH values and concentration level
(mg/l) of various bioactive compounds (Alkaloids) are given in table 5.Which
shows that bioactive compounds observed in higher amount in these valuable
plants and could be used against various infections and diseases.
������������� The data obtained about various
compounds found in roots and leaves of Justicia
adhatoda are given in the tables 3 and 4.While,
comparison of these chemical compounds are presented in figure 2. It was
observed that roots of Justicia adhatoda contained higher concentration of protein fat
and alkaloids like vasicine and vasicinone.(Table 3 ).The leaves of Justicia
adhatoda contained higher concentration of sugar
and vitamin C ( Table 4 ).
����������� It was observed that roots, leaves
and fruit of both plant species contained higher concentrations of chemicals
those can be used against various disorders in human population. The extract of
roots and leaves of Justicia adhatoda are commonly used by rural population against
diabetes, cough and certain liver disorders.�
( Sivarajan and Balachandran, 1994 ).
����������� The roots of Berberis
lyceum are commonly used by people for their body pain to repair cut,
wounds and also against high grade fever (Ivanovska
and Philipov,1996). Similarly fruits of this plant
also have various medicinal values ( Chopra , 1998 ).
Table 1.��
Analysis of Bioactive compounds from roots of Berberis
lyceum.
Constituent
|
Percentage
|
Constituent
|
Percentage
|
Dry matter |
61.2 |
Calcium |
2.2 |
Moisture |
20.5 |
Sodium |
1.5 |
Protein |
4.5 |
|
0.2 |
Fat |
2.6 |
Iron |
0.3 |
��
Sugar |
3.5 |
Zinc |
0.2 |
Fiber |
2.5 |
Berberine |
4.5 |
Palmatine |
3.1 |
Vitamin C |
0.3 |
Table 2. ���Analysis of
bioactive compounds from fruits of Berberis
lyceum.
Constituent
|
Percentage
|
Constituent
|
Percentage |
Dry
matter |
62.5 |
Calcium |
1.8 |
Moisture |
12.5 |
Sodium |
0.6 |
Protein |
2.5 |
|
0.1 |
Fat |
1.8 |
Iron |
0.2 |
Sugar |
4.5 |
Zinc |
0.8 |
Fiber |
1.5 |
Berberine |
2.9 |
|
|
Vitamin
C |
0.8 |
Table 3.� Analysis of Bioactive compounds from roots of Justicia
adhatoda.
Constituent |
Percentage |
Constituent |
Percentage |
Dry matter |
66.4 |
Calcium |
3.1 |
Moisture |
24.6 |
Sodium |
2.4 |
Protein |
8.5 |
|
1.2 |
Fat |
2.5 |
Iron |
0.7 |
Sugar |
2.6 |
Zinc |
0.5 |
Fiber |
5.2 |
Berberine |
0.3 |
Vasicine |
7.5 |
Vitamin C |
5.2 |
Table 4. Analysis of Bioactive compounds from leaves of Justicia adhatoda.
Constituent |
Percentage |
Constituent |
Percentage |
Dry matter |
50.4 |
Calcium |
1.5 |
Moisture |
15.3 |
Sodium |
1.4 |
Protein |
6.5 |
|
1.3 |
Fat |
1.6 |
Iron |
1.2 |
Sugar |
16.4 |
Zinc |
0.6 |
Fiber |
6.4 |
Vasicinone |
3.5 |
Vasicine |
4.5 |
Vitamin C |
1.5 |
Table�� 5. Spectrophotometric analysis of various alkaloids
at 470nm and their PH values.
Alkaloids |
Concenteration |
PH |
1 |
0.724 |
7.8 |
2 |
0.668 |
7.9 |
3 |
0.716 |
7.7 |
4 |
0.163 |
6.9 |
5 |
0.113 |
7.0 |
6 |
0.097 |
6.9 |
7 |
0.58 |
7.2 |
8 |
0.51 |
6.7 |
9 |
0.95 |
7.6 |
Figure
1:�
���Comparison of bioactive compounds in roots
and fruits of Berberis lyceum.
Figure 2:������ Comparison of bioactive compounds in
roots and leaves of Justicia adhatoda.
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