Journal of Pharmacognosy and Phytochemistry 2014; 3 (1): 14-17

ISSN 2278-4136
ISSN 2349-8234
JPP 2014; 3 (1): 14-17 Phenolic Compound, Free radical assay, Anti-microbial
Received: 13-03-2014
Accepted: 16-04-2014
and Anti-fungal Investigation of Pterospermum
Md. Taraquzzaman
semisagittatum: A Herbal Flora of Bangladesh
Department of Pharmacy, State
University of Bangladesh, Dhaka-
1205, Bangladesh. Md. Taraquzzaman, Md. Nur Alam, Arshida Zaman Boby, Faisal Asif, Md.
E- mail: [email protected] Ashraful Islam and Md. Al Amin Sikder
Md. Nur Alam
Department of Pharmacy, State ABSTRACT
University of Bangladesh, Dhaka- The present study was on Pterospermum semisagittatum investigated for its phenolic content, free radical
1205, Bangladesh. scavenging (DPPH) assay, Antimicrobial and fungal activity. The highest phenolic content was found in
E- mail: [email protected]
the pet ether (PESF) (4.25±0.08 mg) followed by carbon tetrachloride soluble partitionate (CTCSF)
Arshida Zaman Boby (2.70±0.27 mg), chloroform soluble partitionate (CSF) (0.56±0.29 mg), methanolic extract (ME)
Department of Pharmacy, State (2.41±0.14 mg) and aqueous soluble partitionate (AQSF) (1.67±0.06 mg). In free radical scavenging
University of Bangladesh, Dhaka-1205, (DPPH) assay, methanolic extract (ME), carbon tetrachloride soluble partitionate (CTCSF), pet ether
Bangladesh. (PESF), chloroform soluble partitionate (CSF) and aqueous soluble partitionate AQSF respectively, IC 50
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was 121.50±0.71, 112.50±1.14, 102.50±0.65 and 53.50±2.12. In Anti-microbial and anti-fungal
Md. Ashraful Islam investigation only chloroform soluble partitionate observed the Slight amount of activity. This study
Department of Pharmacy, State profound that Pterospermum semisagittatum has the remarkable presence of phenolic and antioxidant
University of Bangladesh, Dhaka- compound on preliminary investigational evaluation.
1205, Bangladesh.
E- mail: [email protected] Keywords: Pterospermum semisagittatum, Antioxidant, DPPH, Phenolic Content, Anti-microbial activity,
Anti-fungal activity.
Md. Al Amin Sikder
Department of Pharmaceutical
Chemistry, Faculty of Pharmacy,
1. Introduction
University of Dhaka, Dhaka-1000, Medicinal plants have bestowed a good deal of arising new medicinal molecules. Scientist look
Bangladesh around plants, herbs, microorganisms, marine resources etc. to incorporate new lead
E- mail: [email protected] compounds. Bangladesh is blessed with numerous medicinal plants used for traditional healing,
but very few are light of research [1]. In plants antioxidant activity plays a significant role in
biological molecules brings about to, hydroxyl radicals, superoxide, nitric acid and various
reactive oxygen varieties (ROS) i.e. hypochloric acid, hydrogen peroxide, and proxynitrite[2,3].
Also Phenolic intensifies are widely distributed in the plant kingdom. Plant secondary lipid
precursors and aromatic amino acids. Phenolic compounds build an important portion of the
secondary plant compounds. Lignin, a complex polymer of phenylpropane units is,
quantitatively, the most important phenolic compound in plants. Other abundant compound
classes are the flavonoids, coumarins, stilbenes and polyflavonoids. Now a day scientist has
investigate medicinal plants for new antimicrobial activities [4-5].
The flora for example Pterospermum semisagittatum is endogenous in Bangladesh. This plant
widely used in Herbal medicine as for gastrointestinal disorders, respiratory tract disorders, skin
disorders, burning sensations, hepatic disorders, malaria, rheumatic pain cancer, tumor and heart
palpitation etc. [6]. In these Study, the antioxidant, poly phenol and antimicrobial activity of
Pterospermum semisagittatum was examined.

2. Materials and Methods

2.1 Plant collection
Correspondence: The leaves of Pterospermum semisagittatum (family Sterculiaceae) were accumulated from
Faisal Asif Dhaka, Bangladesh area. A voucher specimen (DACB-39205) has been banked in the National
Department of Pharmacy, State Herbarium, Dhaka.
University of Bangladesh, Dhaka-
1205, Bangladesh.
E- mail: [email protected]

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 Journal of Pharmacognosy and Phytochemistry

2.2 Reagents and chemicals the control, and A1 is the absorbance of the extract/ standard. Then
All chemicals, i.e. methanol, n-hexane, carbon tetrachloride, the inhibition curves were prepared and IC50 values were
chloroform, Gallic acid, 1, 1- diphenyl-2- picrylhydrazyl (DPPH), calculated. BHT was used as positive control.
Folin-Ciocalteu reagent, butylated hydroxytoluene (BHT), ascorbic
acid and other reagents used in these experiments were of the 2.6 The antimicrobial and fungal Investigation
highest analytical grade. The bacterial and fungal strains used for the experiment were
collected as pure cultures from the Institute of Nutrition and Food
2.3 plant Extraction and Isolation Science (INFS), University of Dhaka. Standard disc of
The sun dried powdered leaves approximately 500 gm were Ciprofloxacin (30 μg/disc) and blank discs impregnated with
macerated in 1.5 L of methanol for a week. The extract was filtered solvents followed by evaporation were used as positive and
a through fresh cotton bed and finally with Whatman filter paper negative control. Both gram positive, gram-negative and fungal
number 1 and concentrated with a rotary evaporator at reduced activity were taken for the test and they are listed in the table 2 the
temperature and pressure. An aliquot (5 gm) of the concentrated test material having antimicrobial activity and fungal activity
methanol extract was fractionated [7]. Partition protocol and the inhibited the growth of the microorganisms and a clear, distinct
resultant were evaporated to dryness with a rotary evaporator to zone of inhibition was visualized surrounding the discs. The
yield to which petroleum ether (650.0 mg), carbon tetrachloride antimicrobial activity of the test agents was determined by
(950.0 mg), chloroform (450.0 mg) and aqueous (2.05 gm) soluble measuring the diameter of the zone of inhibition expressed in mm.
material from the crude methanol extract of leaves. All method as following Ayafor et al. (1972) and Bauer et al.
(1966) [11, 12].
2.4 Phenolic Content Investigation
Total phenolic content of Pterospermum semisagittatum leaf 3. Result and Discussion
extracts was assessed by using the method Demiary et al. (2009) The methanol extract of leaf of Pterospermum semisagittatum and
and Skerget et al. (2005)[8,9]. Necessitating gallic acid as a standard its different partitionates i.e. aqueous (AQSF), chloroform (CSF),
and Folin-Ciocalteu reagent as an oxidizing agent. To 0.5 ml of carbon tetrachloride (CTCSF), and soluble partitionates pet ether
extract solution (2 mg/ml) in water, 2.5 ml of Folin-Ciocalteu (PESF) were subjected to total phenolic activity, Free radical
reagent (10 times diluted with water) and 2.0 ml of sodium Scavenging activity and Anti-microbial activity. Based on the
carbonate (7.5 % w/v) solution were added. After 20 minutes of absorbance values of the various extract solutions, reacted with
incubation at 250C and the absorbance was assessed using a UV Folin-Ciocalteu reagent and Calculated with the standard solutions
visible spectrophotometer at 760 nm. Total phenolics were of gallic acid curve (R2=0.9985).The colorimetric analysis of the
quantified by calibration curve obtained from the known total phenolic contents are acceded Figure 1 and As well table
concentrations of gallic acid (0-100 μg/ml) and were expressed as 1.Among all extractives of Pterospermum semisagittatum, the
gm of GAE (gallic acid equivalent) / 100 gm of the dehydrated highest phenolic content was found in PESF (4.25±0.08 mg of
extract. GAE / gm of dried extracts) followed by CTCSF (2.70±0.27 mg of
GAE / gm of dried extracts). A significant amount of phenolic
2.5 Free radical scavenging (DPPH) Investigation compounds were present in CSF (0.56±0.29 mg of GAE / gm of
The free radical scavenging activity of the extract, based on the dried extracts), ME (2.41±0.14 mg of GAE / gm of dried extracts)
scavenging activity of the stable 1, 1- diphenyl-2- picrylhydrazyl and AQSF (1.67±0.06 mg of GAE / gm of dried extracts).
(DPPH) free radical, was determined by the method described by In free radical scavenging (DPPH) assay, methanolic extract (ME),
Braca et al. (2001) [10]. 2.0 ml of a methanol solution of the extract Carbon tetrachloride soluble partitionate (CTCSF), pet ether
at different concentration (500 to 0.977 μg ml) were mixed with 3.0 (PESF), chloroform soluble partitionate (CSF) and aqueous soluble
ml of a DPPH methanol solution (20μg/ml). After 30 minutes partitionate AQSF respectively, for IC50 was 121.50±0.71,
reaction period at room temperature in a dark place the absorbance 112.50±1.14, 102.50±0.65 and 53.50±2.12. In comparison to the
was measured against at 517 nm against methanol as blank by UV standard tert-butyl-1- hydroxytoluene (BHT) (IC50 27.75±0.35) and
spectrophotometer. The percentage inhibition activity was ascorbic acid (ASA) (IC50 5.95±0.21) in Table 1. The percent of
calculated from [(A0–A1)/A0] ×100, where A0 is the absorbance of inhibition versus Concentration was plotted in Figure 2.

Fig 1: Total phenolic content various dried extracts

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 Journal of Pharmacognosy and Phytochemistry

Table 1: Total Phenolic content of various dried extracts and IC50 Value of In vitro Free radical scavenging (DPPH) activity of
Pterospermum semisagittatum
Total Phenolic Content (mg of Free Radical Scavenging
Name of the Samples
GAE/gm of dried extract) (DPPH) Value of IC50
AQSF 1.67±0.06 27.75±1.06
CSF 0.56±0.29 53.50±2.12
CTCSF 2.70±0.27 112.50±1.14
PESF 4.25±0.08 102.50±0.65
ME 2.41±0.14 121.50±0.71
BHT NA 27.75±0.35
ASA NA 5.95±0.21
AQSF= aqueous soluble fraction,CSF= chloroform soluble fraction, CTSF= carbon tetrachloride soluble fraction, PESF= pet ether soluble
fraction, ME= methanol extract, BHT = Butylated hydroxytoluene, ASA = Ascorbic Acid and NA = Not Analyzed. All calculation is
performed with triplicate mean± Standard Deviation.

Fig 2: Comparative DPPH radical scavenging activity of different extractive of Pterospermum semisagittatum leaves along with BHT and
Ascorbic acid including Mean ± Standard deviation

Table 2: Antimicrobial activity of test samples of Pterospermum semisagittatum

Diameter of zone of inhibition (mm)
Test microorganisms
AQSF CSF CTCSF PESF ME Ciprofloxacin
Gram positive bacteria
Bacillus cereus - 10 - - - 40
Bacillus megaterium - 14 - - - 50
Bacillus subtilis - 8 - - - 36
Staphylococcus aureus - - - - - 42
Sarcina lutea - 10 - - - 42
Gram negative bacteria
Escherichia coli - - - - - 50
Pseudomonas aureus - - - - - 49
Salmonella paratyphi - 8 - - - 37
Salmonella typhi - 12 - - - 35
Shigella boydii - - - - - 45
Shigella dysenteriae - 10 - - 40
Vibrio mimicus - - - - - 42
Vibrio parahaemolyticus - 12 - - - 36
Fungi
Candida albicans - - - - - 35
Aspergillus niger - 8 - - - 34
Saccharomyces cerevisiae - - - - - 45
AQSF= aqueous soluble fraction, CSF= chloroform soluble fraction, CTSF= carbon tetrachloride soluble fraction, PESF= pet ether soluble fraction,
ME= methanol extract

The exploratory antimicrobial activity of the extracts was number of Gram positive and Gram negative bacteria and fungi.
determined at 400 μg/disc by the disc diffusion method against a The results are given in the Table 2.
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During the measurement of zone of inhibition, only a chloroform Morelli I et al. Antioxidant principles from Bauhinia
soluble fraction has the slight amount of antimicrobial and tarapotensis. Journal of Natural Products 2009; 64(7):892-
antifungal activity. Highest amount of activity shown against 895
Bacillus megaterium (14 mm), Salmonella tophi (12 mm),Vibrio 11. Ayafor JF, Kimbu SF, Ngadjui BT. Limonoids and phytol
parahaemolyticus (12mm), Bacillus cereus (10 mm), Sarcina lutea derivatives from Cedrela sinensis. Tetrahedron 1972;
(10 mm), Shigella dysenteriae (10 mm), Bacillus subtilis (8 mm), (28):9343
Salmonella paratyphi (8 mm) and antifungal activity Aspergillus 12. Bauer AW, Kirby WMM, Sherries JC, Tuck M. Antibiotic
niger (8 mm) etc. But the comparative Standard ciprofloxacin is susceptibility testing by a standardized disc diffusion
much higher active than Pterospermum semisagittatum. method. Ame J Clin Pathol 1966; 45:493-496.

4. Conclusion
The results of Phenolic content and Free radical scavenging
(DPPH) were relatively significant. Further more focused research
on this plant is recommended. Scientists can obtain more
significant medicinal compounds from this plant.

5. Acknowledgement
The authors wish to best regards for the phytochemical research
laboratory of State University of Bangladesh and Institute of
Nutrition and Food Science (INFS), University of Dhaka,
Bangladesh for supplying test organisms to perform these
investigations.

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