Antimicrobial activity and pharmacognostic studies of the flowers of Butea monosperma
My Research Artcicle published
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Research Article
Pharmacognostical studies on Butea monosperma (Lam.) Taub (Faboideae)
flower
Shruti V. Hegde*, G. R. Hegde, Shruti Mannur, Shreedevi S. Poti
Post graduate Department of studies in Botany, Karnatak University, Dharwad- 580 003,
Karnataka.
Abstract:
Butea monosperma, a deciduous tree belonging to the family Faboideae, is found growing in many
parts of India. All the parts of plant are highly medicinal with its mention in different systems of
medicine. Several review works have summarized the potential efficiency of this plant. This
Pharmacognostic study comprises taxonomic details, macro and microscopic characters, physicochemical
details and study of phytochemical components of all five successive extracts. The work is
done for pharmacognostic standardization and authentication of flowers of Butea monosperma.
Keywords: Butea monosperma , Pharmacognosy, Phytochemical analysis
1. Introduction:
Butea monosperma (Lam.) Taub (Syn. Butea frondosa Willd. Family Faboideae), a
deciduous tree, is found chiefly in the mixed or dry deciduous forests of Central and Western
India. This plant is popularly known as dhak or palas, palash, mutthuga, bijasneha, khakara,
chichara and commonly known as „Flame of the forest‟. This tree grows to 50 ft high, with
stunning flower clusters. Tree is almost leafless during spring season forming an orange- red
hue of flowers on the upper portion, giving the appearance of flame from a distance 1,2.
B. monosperma is extensively used in Ayurveda, Unani, Homeopathy and Traditional
systems of medicine. Flowers of B. monosperma are used as anticonvulsant, antioxidant,
antistress, antigout, diuretic, antileprotic, anti-inflammatory, antiulcer, astringent,
ISSN (Online) 2249-6084 (Print) 2250-1029
International Journal of Pharmaceutical and
Phytopharmacological Research (eIJPPR)
[Impact Factor – 0.7826]
Journal Homepage: www.eijppr.com
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antiestrogenic activity, antihepatotoxic, eye disorder 2,3, diarrhea4, depurative, tonic, leprosy, skin diseases and thirst5. Phytochemical studies of flower extract have shown chemical constituents like triterpene, flavonoids and glycosides like butein, butin, isobutrin, coreopsin, isocoreopsin, sulphurein, monospermoside, isomonospermoside, chalcones, aurones and steroids6,7,8.
Each plant drug possesses unique properties in terms of its botany, chemical constituents and therapeutic potency. So it is important to study pharmacognostic characters of each medicinal plant to differentiate the genuine plant sample. Isolation and pharmacological studies have been extensively made on all parts of B. monospema but, very less is known about pharmacognosy. Present work is to frame a standard Pharmacognostic parameters for the flowers of Butea monosperma useful in authentification and standardization of the drug, which can guarantee the quality and purity of the drug.
2. Materials and Methods
2.1 Plant material:
Flowers of B. monosperma were collected from the Halligeri village, Dharwad District, Karnataka. The plant was authentically identified using Flora of Karnataka9. The recent name has been given based on IPNI10. Herbarium specimen (Voucher specimen number: KUD/BOT/2012/10) was prepared and deposited in Department of Botany, Karnatak University, Dharwad, along with powder samples (KUD/BOT/2012/10) for future reference study.
The collected flowers were cleaned and shade dried. Fresh samples were used for anatomical studies and dried parts were powdered, sieved and stored in an airtight container for further use.
2.2 Macroscopic and Microscopic analysis:
Key morphological features were observed for easy identification. Microscopic studies were carried out by using dissecting microscope (AJAY® OPTIK INDI: AJ-2. CM/L-9018771). Powder studies were carried out by using reagents and stains like iodine, potassium iodide, ferric chloride, Sudan III, ruthenium red and phloroglucinol with Con. HCl (1:1) 11,12,13. Safranin (4%) and toludine blue were used to double stain the transverse sections 11,14. All the reagents of analytical grade were procured from Hi-Media, Mumbai, India. Organoleptic characters like colour, texture, odor and taste were determined for flower powder 11.
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2.3 Photo documentation:
Photomicrographs of free hand sections and powder microscopy were taken using compound binocular microscope at different magnifications (Carl Zeiss Axio Imager M2 model) with inbuilt analogue camera (ProgRess C5- JENOPTIK). Computer images were captured using ProgRes® CapturePro 2.8- JENOPTIK optical system software.
2.4 Physico-chemical analysis:
Physico-chemical parameters of the powdered drug such as total ash, water-soluble ash, acid-insoluble ash and sulphated ash were determined. Extractive value, solubility tests, moisture test, mineral content and nutritive value (ash, fat, fiber, protein and carbohydrate) of flowers were determined as per standard procedures15,16. Foaming index, bulk and tapped density, hausner ratio and carr index, swelling index, moisture sorption capacity, pH and hydration capacity of the powder sample13,17 were studied.
2.5 Calculation of % carbohydrate and nutritive value:
Percentage of carbohydrate was calculated by the following formula:
% carbohydrates = 100 - (Percentage of ash + percentage of moisture + percentage of fat + percentage of protein)
Nutritive value was finally determined by:
Nutritive value = 4 x percentage of protein + 9 x percentage of fat + 4 x percentage of carbohydrate
2.6 Fluorescence analysis:
The treated powdered sample materials of flower were analyzed under visible light, short ultra-violet light (254nm) and long ultra-violet light (365nm)18.
2.7 Preparation of extracts and preliminary phytochemical analysis:
The powdered material was serially extracted by Soxhlet extraction method using hexane, chloroform, acetone, ethanol and water. These extracts were subjected for preliminary phytochemical screening 19.
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2.8 Data analysis
Standard deviation is calculated as mean of three replicates for flower constants and physico-chemical parameters using SPSS version 16.0, statistical package. Data is represented in tables.
3. RESULTS
3.1 Macroscopic characters of flower:
Flowers large, in rigid racemes, up to 15 cm long, 3 flowers together from the tumid nodes of the dark olive-green velvety rachis; pedicels as long as the calyx, densely brown-velvety; bracts and bracteoles small, deciduous (Plate 1. Fig. A, B and D). Calyx 1.3 cm long, dark olive- green, densely velvety outside, clothed with silky hairs within; teeth short, the 2 upper connate, the 3 lower equal, deltoid. Corolla 3-5 cm long, clothed outside with silky silvery hairs, orange or salmon coloured; standard 2.5 cm broad; keel semicircular, beaked, veined. Stamens 10, monoadelphous, basifixed glabrous filament. Ovary superior, style hairy, stigma globular hairy and monocarpellary unilocular.
3.2 Anatomical characters of Flower: Olive green coloured calyx section shows oval shaped epidermal cells subtended by brownish unicellular trichomes, oil ducts and internally 6-7 layered cortical cells (Plate 1. Fig. C, E and F). Corolla shows uni and pitted like multicellular trichomes (Plate 2. Fig. G, H), multilayered parenchyma cells with orange yellow pigments (Plate 2. Fig. I) followed by conjoint, collateral vascular bundle (Plate 2. Fig. J). Inner ovary section shows outer unicellular trichomes (Plate 2. Fig. L), multilayered cortical cells followed by conjoint, collateral and closed vascular bundle (Plate 2. Fig. K, M).
Powder microscopy showed uni and multi cellular trichomes of calyx and corolla. Numerous orange yellow pigmented cells, oil globules, triangular and oval shaped pollens were observed.
3.3 Organoleptic characters:
Powder is yellowish brown, velvety in texture, bitter in taste and smells chocolaty.
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3.4 Physicochemical parameters:
Physicochemical characters such as ash value, mineral content and nutritive value indicated the amount of inorganic constituents (Table 2). Yield is calculated using soluble extractive value and extractive value. Foaming index, bulk and tapped density, hausner ratio, carr index, swelling index, hydration capacity, moisture sorption capacity helps to know moisture content and deterioration time. The pH of sample was slightly basic (Table 1)
3.5 Fluorescence analysis:
Consistency, color and fluorescence activity of powdered drug observed at 254, 365nm and visible light are given in Table 3.
3.6 Phytochemical analysis: The extractive value for successive extracts taken in hexane, chloroform, acetone, alcohol and water were calculated (Table 1). The extract showed sticky nature for all solvents and color difference in visible light (Table 1). All the extracts were subjected to preliminary phytochemical screening and the results certified the presence of alkaloids, phenols, flavonoids, triterpinoids, steroids, carbohydrates, proteins and saponins in alcohol and water extracts of flower.
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Table 1: Physicochemical observations of B. monosperma flower:
Parameter
1. Ash Value (% w/w):
Total ash
2.50 ± 0.02
Acid insoluble ash
1.98 ± 0.01
Water soluble ash
0.78 ± 0.01
Sulphated ash
3.47 ± 0.03
2. Extractive value (% w/w):
Hexane
0.78 ± 0.02
(dark yellowish to orange)
Chloroform
0.59 ± 0.01 (orange)
Acetone
2.11 ± 0.02 (Dark orange)
Alcohol
0.55 ± 0.03 (dark orange)
Water
17.15 ± 0.13 (brownish yellow)
3. Solubility Test (% w/w):
Alcohol
0.60 ± 0.02
Water
1.26 ± 0.01
4. Nutritive content (%)
Ash
2.50 ± 0.02
Moisture
2.00 ± 0.02
Fat
3.14 ± 0.00
Fiber
2.10 ± 0.01
Protein
16.87 ± 0.01
Carbohydrate
75.11 ± 0.01
5. Nutritive value in cal./100 g powder
396.21
6. Foaming index
< 100
7. Swelling index %
46
8. Bulk and Tapped density g/ml
0.55
9. Moisture sorption capacity/g
1.39
10. Hydration capacity g/g
2.309
11. Hausner ratio
1.00
12. Carr index %
20
13. pH
7.69
Table: 2. Mineral content in flowers of B. monosperma.
Mineral content
N %
P %
K %
Na %
S %
Ca %
Mg %
Fe ppm
Mn ppm
Zn ppm
Cu ppm
quantity
2.7
0.27
1.98
0.06
0.1
1.02
0.42
132.83
21.47
68.58
18.18
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Table 3: Ultra- violet powder analysis of flowers of B. monosperma at 245 and 365 nm
Treatment
Visible light
U. V. light at 254nm
U.V. Light 365nm
Flower
Flower
Flower
P + NaOH (Aqueous)
Apache yellow
Door country green
Door country green
P + NaOH (Alcoholic)
International red
Door country green
Light green
P + 1N HCl
Canary yellow
Apple green
Apple green
P + 50% H2SO4
International orange
Apple green
Apple green
P + 50% HNO3
Canary yellow
Light yellow
Chrome yellow
P + Lead acetate + NaOH
International red
Green
Brown
P + HNO3
Canary yellow
Light yellow
Light yellow
P + acetic acid
Canary yellow
Apple green
Apple green
P + FeCl3
Brown
Green
Green
P + HNO3 + NH3
International red
Green
Green
P + H2SO4
Orange
Door country green
Door country green
P + Pet. Ether
Beige color
Light green
Light almond
P + Methanol
Canary yellow
Light green
Light almond
P + Water
Canary yellow
Light green
Light almond
P + Benzene
Canary yellow
Light green
Light almond
P + Glycerin
Canary yellow
Light green
Light almond
P: Powder
4. DISCUSSION: Plants are treasures of medicine. When the plants sound strong traditional significance they are exploited to study their efficacy. Review works by various authors1,3,5,6,7,8 have documented the uses of B. monosperma in different systems of medicine. Their study helped to know the detailed chemical constituents of drug part and potency of plants in pharmacological field, with relevant references. The documented compounds belong to triterpenoids, steroids, carbohydrates, proteins and flavonoids. Present pharmacognostic study of B. monosperma flower is supportive to know the basic characters of the drug like detailed anatomy, physico-chemical parameters, phytochemical constituents, mineral and nutritive value. Some salient features of B. monosperma flowers studied using pharmacognostic features are discussed in this paper. A pigmented bright orange yellow parenchyma cell and pitted multicellular trichomes are salient feature of the drug. Humidity in the sample and extract decides the deterioration time.
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High water content in powder and aqueous extract are found to get deteriorated due to fungal attack. Loss in weight of flower powder on drying at 1050 C was found to be 2.00 %. Analytical results like total ash value was 2.50% which indicate the amount of minerals present in the flower sample. The amount of acid-insoluble siliceous matter was higher than water-soluble ash (1.98%). Compared to solublility value, extractive value of water was higher. Foaming index indicated the presence of saponins in sample. Less value of hausner ratio, carr index, bulk and tapped density indicated good flow. Positive results for alkaloids, saponins and phenols indicate the need of further studies regarding isolation and characterization of these active principle constituents.
5. CONCLUSION
The parameters studied can be utilized in identification of Butea monosperma in crude drug form and can be used as a potential source for useful therapeutics. The resulted data will be beneficial for quantitative and qualitative standardization of genuine drug in herbal preparations. Positive result for alkaloids, saponins and phenols is indicative of scope for future analysis.
6. ACKNOWLEDGEMENTS:
Authors are grateful to UGC, New Delhi, India for the financial assistance and Karnatak University, Dharwad for the facilities provided. Authors extend their sincere thanks to USIC, Karnatak University, Dharwad for Instrumental support.
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