Phytochemical and Pharmacological Evaluation of Ethanolic Extract of Thorns of Mangrove Plant Species-Phoenix Paludosa Roxb
Phytochemical and Pharmacological Evaluation of Ethanolic Extract of Thorns of Mangrove Plant Species-Phoenix Paludosa Roxb
Phytochemical and Pharmacological Evaluation of Ethanolic Extract of Thorns of Mangrove Plant Species-Phoenix Paludosa Roxb
Presented By:
Examination Roll No.: 131027
Examination Session: 2013-2014
Phoenix paludosa Roxb
Department of Pharmacy
Jashore University of Science and Technology.
Jashore-7408, Bangladesh
Plant Preview
Phoenix paludosa Roxb
Family: Arecaceae
Description:
Perennial palm growing up to a height of 6 m. Stem slender, cylindrical,
unbranched, distinct leaf scars. Leaves pinnately compound, waxy,
glabrous, midrib strong, ending into strong sharp spine at apex, few
pairs of lower segments modified into sharp spines.
Distribution:
Andaman Is., Assam, Bangladesh, Cambodia, India, Malaya, Myanmar,
Nicobar Is., Sumatera, Thailand, Vietnam.
Traditional Uses
Anti-helminthic, Whole plant is used in construction of house, used as
medicine for insomnia.
Literature Review of Phoenix paludosa Roxb
Findings Plant parts Reference
Lima, A., Parial, R., Das, M., & Das, A. K. (2010). Phytochemical and
Phytochemical and pharmacological studies of ethanolic extract from the leaf of mangrove
pharmacological studies Leaves plant Phoenix paludosa Roxb. Malaysian Journal of Pharmaceutical
Sciences, 8(2), 59-69.
Saha, S., Islam, M. K., Anisuzzman, M., Hasan, M. M., Hossain, F., &
antioxidant, analgesic and Leaves Talukder, C. (2012). Evaluation of antioxidant, analgesic and antidiarrheal
antidiarrheal activity activity of Phoenix paludosa roxb Leaves. International Journal of Basic
Medical Sciences and Pharmacy (IJBMSP), 2.
Development and Akkak, A., Scariot, V., Marinoni, D. T., Boccacci, P., Beltramo, C., & Botta, R.
(2009). Development and evaluation of microsatellite markers in Phoenix
evaluation of microsatellite Leaves
markers dactylifera L. and their transferability to other Phoenix species. Biologia
Plantarum, 53(1), 164-166.
Aims and Objectives
So the leaves of Phoenix paludosa were selected for the present study that was arranged as follows.
A: Phytochemical tests: To identify the presence or absence of different phytochemical groups.
B: Pharmacological assessment: To evaluate different pharmacological activities like Phytochemical
Screening, Antioxidant activity, Blood coagulation activity, Blood anti-coagulation activity, Anti-
hyperglycemic activity.
Rotary
Collection Filtration
Evaporation
Vacuum
Chopping Maceration desiccation
60.00
50.00
40.00
30.00 Ascorbic acid 77.83
20.00
10.00
0.00
1 2 4 8 16 32 64 128 256 512 1024
P. paludosa extract 181.02
Concentration
Standard P. paludosa
Finding: P. paludosa activity of DPPH free radical scavenging is comparable with Ascorbic acid
Sadhu, S.K., Okuyama, E., Fujimoto, H., Ishibashi, M., 2003. Chemical and Pharmaceutical Bulletin 51, 595-598.
Evaluation of Antioxidant Activity (Cont.)
Hydrogen Peroxide Radical Scavenging Assay
Different concentrations (6.25 to 800 μg/ml) of extracts were added to a H 2O2 solution (40 mM) and at 230
nm absorbance was determined after ten minutes to obtain SC50 values (Keser et al., 2012).
% Scavenged vs Concentration
120.00
Result:
100.00
80.00
Test Sample
% Scavenged
SC50 (µg/ml)
60.00
40.00
Ascorbic acid 89.63
20.00
0.00
6.25 12.5 25 50 100 200 400 800 P. paludosa extract 107.59
Concentration
Standard P. Paludosa
Finding: P. paludosa activity of H2O2 radical scavenging is comparable with ascorbic acid
Keser, S., Celik, S., Turkoglu, S., Yilmaz, O., Turkoglu, I., 2012. Chemistry Journal 2, 9-12.
Evaluation of Antioxidant Activity (Cont.)
Determination of Total Phenolic Content (TPC)
Total phenolics of plant extracts were measured using Folin-Ciocalteu reagent and by evaluating the
regression equation of the calibration curve (y = mx + c), expressed in mg gallic acid equivalents (GAE) per
gram of dry powder (mg GAE/g) (Javanmardi et al., 2003).
Absorbance of Gallic Acid
Absorbance Linear (Absorbance)
Result:
Absorbance
1
0.77
0.8
f(x) = 5.55 x − 0.03 0.63 Total phenolic content
0.6
R² = 0.95 Sample
(mg GAE/g)
0.35
0.4 0.3
0.18
0.2 0.09 P. paludosa 451.63
0
0 0.02 0.04 0.06 0.08 0.1 0.12 0.14 0.16
Concentration (mg/ml)
0.5
Result:
0.42
0.4 f(x) = 0.42 x + 0.03
R² = 0.99 0.34
0.27
Total flavonoid content
Absorbance
0.3 Sample
(mg QE/g)
0.2
0.13
0.1
0.01
P. paludosa 89.05
0
0 0.25 0.5 0.75 1
Concentration (mg/ml)
P. paludosa 280.50
0
0 1 2 3 4 5 6 7
Concentration (mg/ml)
1.2
1
0.92 Sample
0.8 0.67
(mg AAE/g)
0.6
0.4
0.2 0
0.23
P. paludosa 249.59
0
0 0.1 0.2 0.3 0.4 0.5 0.6
Concentration (mg/ml)
50 48.48
43.92
40
Coagulation Time (min)
36.88
Fresh Blood
30 1.25 mg/ml 27.75
100 mg/ml
200 mg/ml
10 mg/ml
50 mg/ml
5 mg/ml
25 mg/ml
mg/ml
20
2.5
10 7.45 8.57
4.35 5.45
2.21
0
Phytomenadione vit k1 P. paludosa Negative Control
Figure: Effect of extract on clotting time (min) of blood in a blood coagulation activity test.
Result: P. paludosa extract, further evaluation was carried out to evaluate the in vitro blood anti-coagulation
activity.
Ikese, C., Okoye, Z., Kukwa, D., Adoga, S., Lenka, J., 2015. International Journal of Pharmaceutical Sciences and Research 6, 3391.
Evaluation of in vitro Blood Anticoagulation Activity
The anticoagulant activity was evaluated by prothrombin time test. Blood samples were obtained from
normal individuals and pure platelet plasma (PPP) was isolated by centrifugation. Plasma, plant extracts
and Cacl2 were added together and clotting time was recorded with stop watch (Mao et al., 2009).
58.75
350 mg/ml
60
175 mg/ml
50
Coagulation Time (min)
87.5 mg/ml
0.9% saline
40
5 mg/ml
33.88
30
24.9
20
13.5
10 8.54
0
warfarin P. peludosa Control
Figure: Effect of extract on clotting time (min) of blood in a blood anticoagulation activity test.
Result: P. paludosa extract exhibited considerable and significant blood anticoagulant activity.
Mao, W., Li, H., Li, Y., Zhang, H., Qi, X., Sun, H., Chen, Y., Guo, S., 2009. International Journal of Biological Macromolecules 44, 70-74.
Evaluation of Antihyperglycemic Activity (Cont.)
In vitro Evaluation of α-Glucosidase Enzyme Inhibitory Activity
Mixture of potassium phosphate buffer, enzyme solution and sample or standard solution was first incubated.
Then pNPG (substrate) and Na2CO3 solution was added to it and absorbance was measured at 405 nm to obtain
% inhibition Result:
40
20
Test Sample IC50 (mg/ml)
0
0.1 0.2 0.3 0.4 0.5 1
-20
Acarbose 0.382
-40 P. paludosa extract 1.864
-60
-80
Findings: P. paludosa plant extracts exhibited antihyperglycemic activity by inhibition of α-glucosidase enzyme.
Lawag, I.L., Aguinaldo, A.M., Naheed, S., Mosihuzzaman, M., 2012. Journal of Ethnopharmacology 144, 217-219.
Qaisar, M.N., Chaudhary, B.A., Sajid, M.U., Hussain, N., 2014. Tropical Journal of Pharmaceutical Research 13, 1833-1836.
Conclusion
According to the results of the present investigation, following results are
1. The plant extract revealed the presence of reducing sugar, phenolic compound,
tannins, terpenoids, saponins, steroid, flavonoids, alkaloids & glycosides
2. The Plant extract exhibit good antioxidant activity like DPPH free radical
scavenging activity, H2O2 radical scavenging activity, TPC, TFC, TTC, TAC
I wish to express my profound sense of gratitude to all the respectable teachers and other
members of Pharmacy Discipline for their support to carry out this thesis work.