In vitro Phytochemical Screening and Antibacterial Activity of Organic Leaf Extracts of Spathodea campanulata P. Beauv Against Hospital Isolated Bacterial Strains
ABSTRACT Spathodea campanulata P. Beauv is extensively used in Indian traditional and folklore medicines to cure various human ailments. The preliminary phytochemical screening of the leaves revealed the presence of alkaloids, flavonoids, saponins, tannins, steroids and terpenoids. In vitro antibacterial studies of the leaf extracts were carried out on eight medically important bacterial strains using agar disc diffusion method. The antibacterial assay using petroleum ether leaf extracts showed good inhibitory activity against Klebsiella pneumoniae and compared with standard antibiotic Streptomycin.
- Planta Medica 05/1988; 54(2):122-5. · 2.35 Impact Factor
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ABSTRACT: Antimicrobial activity of 18 ethnomedicinal plant extracts were evaluated against nine bacterial strains (Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa, Ervinia sp, Proteus vulgaris) and one fungal strain (Candida albicans). The collected ethnomedicinal plants were used in folk medicine in the treatment of skin diseases, venereal diseases, respiratory problems and nervous disorders. Plants were collected from Palni hills of Southern Western Ghats and the ethnobotanical data were gathered from traditional healers who inhabit the study area. The hexane and methanol extracts were obtained by cold percolation method and the antimicrobial activity was found using paper disc diffusion method. All microorganisms were obtained from Christian Medical College, Vellore, Tamil Nadu, India. The results indicated that out of 18 plants, 10 plants exhibited antimicrobial activity against one or more of the tested microorganisms at three different concentrations of 1.25, 2.5 and 5 mg/disc. Among the plants tested, Acalypha fruticosa, Peltophorum pterocarpum, Toddalia asiatica,Cassia auriculata, Punica granatum and Syzygium lineare were most active. The highest antifungal activity was exhibited by methanol extract of Peltophorum pterocarpum and Punica granatum against Candida albicans. This study evaluated the antimicrobial activity of the some ethnomedicinal plants used in folkloric medicine. Compared to hexane extract, methanol extract showed significant activity against tested organisms. This study also showed that Toddalia asiatica, Syzygium lineare, Acalypha fruticosa and Peltophorum pterocarpum could be potential sources of new antimicrobial agents.BMC Complementary and Alternative Medicine 02/2006; 6:35. · 2.08 Impact Factor
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ABSTRACT: The antimicrobial properties of the hexane extract from the leaves and twigs of the Eupatorium glutinosum was tested against two Gram-negative and two Gram-positive bacteria. Bioassay guided fractionation leads characteristically to isolation and structure elucidation of two active compounds: 15-hydroxy-7-labden-17-oic acid and its acetate. These results validate the vernacular medicinal uses of the plant in folkloric medicine. Dammara-20, 24-dien-3-acetate, stigmasterol and epi-friedelanone have been also isolated in addition to the palmitate of nepehinol for the first time as naturally occurring.Journal of Ethnopharmacology 08/2002; 81(2):293-6. · 2.94 Impact Factor
Ethnobotanical Leaflets 12: 1022-28. 2008.
In vitro Phytochemical Screening and Antibacterial Activity of Organic Leaf
Extracts of Spathodea campanulata P. Beauv against Hospital Isolated Bacterial
*Rangasamy Dhanabalan, Asirvatham Doss, Subbu Balachandar,
Ethiraj Kezia, Muthusamy Jagadeeswari and Haldurai Karthik
Department of Microbiology, Centre for Post Graduate Studies in Microbiology
RVS College of Arts and Science, 242 –B KVK Thottam,Trichy Road,
Coimbatore-641 402, Tamilnadu, India.
*Corresponding author: email@example.com
Issued 17 November 2008
Spathodea campanulata P. Beauv is extensively used in Indian traditional and folklore medicines to cure
various human ailments. The preliminary phytochemical screening of the leaves revealed the presence of alkaloids,
flavonoids, saponins, tannins, steroids and terpenoids. In vitro antibacterial studies of the leaf extracts were carried
out on eight medically important bacterial strains using agar disc diffusion method. The antibacterial assay using
petroleum ether leaf extracts showed good inhibitory activity against Klebsiella pneumoniae and compared with
standard antibiotic Streptomycin.
Key words: Spathodea campanulata, Phytochemical Screening, Leaf extracts and Antibacterial activity.
Infectious diseases are the leading cause of death worldwide. The clinical efficacy of many existing
antibiotics is being threatened by the emergence of multidrug-resistant pathogens (Bandow et al., 2003). Bacterial
and fungal pathogens have evolved numerous defense mechanisms against antimicrobial agents, and resistance to
old and newly produced drugs is on the rise. The increasing failure of chemotherapeutics and antibiotic resistance
exhibited by pathogenic microbial infectious agents has led to the screening of several medicinal plants for their
potential antimicrobial activity (Colombo and Bosisio 1996; Scazzocchio et al., 2001). There are several reports in
the literature regarding the antimicrobial activity of crude extracts prepared form plants (El-Seedi et al., 2002; Rojas
et al., 2003; Duraipandiyan et al., 2006; Parekh and Chanda, 2007a).The use of traditional medicine and medicinal
plants in most developing countries, as a normative basis for the maintenance of good health, has been widely
observed (UNESCO, 1996). Furthermore, an increasing reliance on the use of medicinal plants in the industrialized
societies has been traced to the extraction and development of several drugs, chemotherapeutics from these plants as
well as from traditionally used rural herbal remedies (UNESCO, 1998). The practice of traditional medicine is
widespread in China, India, Japan, Pakistan, Sri Lanka and Thailand. The results of investigation performed in the
late 19 and 20 century and the advent of streptomycin and other antibiotics provide the ground for experimentation
of a vast number of plants for antibiotic or antimicrobial activities that are useful to man (Asirvatham Doss and
Rangasamy Dhanabalan., 2008).
Spathodea campanulata is a species belonging to the Bignoniaceae family, native from equatorial Africa. The
Siddha/Tamil name of this species is Patadi and in folk it is popularly called as Ruugatuuraa. It is very commonly
found and planted in the coffee estates of Munnar, South Tamilnadu and denoted by the name Malaria Maram(tree).
In English the species is called as Syringe tree, Fountain tree, African Tulip tree, Flame-of-the-forest or Nandi
Flame. It is a medium-size tree (15-25 m high), characterized by red garish flowers. It is often employed in
gardening in tropical and subtropical areas including South America (Joly, 1985). Flowers and stem bark extracts
have shown molluscicidal activity and also employed in diuretic, anti-inflammatory treatments. The leaves are used
against kidney diseases, urethra inflammations and as an antidote against animal poisons. The stem bark
preparations are employed against enemas, fungus skin diseases, herpes, stomachaches and diarrhea (Jardim et al.,
2003; Mendes et al., 1986). Hypoglycemic, anti-HIV and antimalarial activities were also observed in stem bark
extracts (Niyonzima et al., 1999; Makinde et al., 1988). Preliminary in vitro antimalarial activity of Spathodea
evaluated (Rangasamy Dhanabalan et al.,2008).
Several phytochemical studies were performed with different parts of S. campanulata, including stem barks,
leaves, flowers and fruits (Ngouela et al., 1990; Ngouela et al., 1988; Amusan et al., 1995; Amusan et al., 1996).
The leaves have furnished spathodol, caffeic acid, other phenolic acids and flavonoids (Ngouela et al., 1991;
Subramanian et al., 1973; El-Hela, 2001a; El-Hela, 2001b). Banerjee and DE (2001) showed the presence of
anthocyanins in flowers of S. campanulata. A qualitative fungitoxic activity of S. campanulata roots against
Cladosporium herbarum CCT 0279 has been evaluated and reported (Pianaro et al., 2007). In vitro antibacterial
activity of leaf extracts of this plant against standard strains was evaluated (Parek.J and Chanda.S, 2007). Thus, the
present work is aimed to evaluate the antibacterial activity of S. campanulata leaf extracts against the clinically
isolated medically important species.
MATERIAL AND METHODS
Fresh plant leaf samples were collected from the Munnar Kundala Tea Estates, Kerala, India during March
2008. The taxonomic identity of the plant was confirmed by Botanical Survey of India (Southern Circle),
Coimbatore,Tamilnadu, India and the voucher specimen of the plants was preserved in Department of Microbiology,
RVS College of Arts and Science, Coimbatore, Tamilnadu, India. Fresh plant material was washed under running tap
water, air dried, homogenized to fine powder and stored in airtight bottles.
Preparation of plant extract
About 100g of the powder was extracted with different organic solvents viz, Benzene, Chloroform, Methanol,
Petroleum ether and Water and allowed to stand overnight. The extract was filtered through Whatman no.1 filter
paper to remove all unextractable matter, including cellular materials and other constitutions that are insoluble in the
extraction solvent. The entire extract was concentrated to dryness using rotary flash evaporator under reduced
The antibacterial activities of the extracts were tested using hospital isolated Gram positive bacteria:
Staphylococcus aureus, Staphylococcus citrus, Bacillus subtilis and Gram negative bacteria: Salmonella typhi,
Pseudomonas aeruginosa, Klebsiella pneumonia, Serratia sp., and Proteus vulgaris. All the strains were procured
from Department of Microbiology, RVS Hospital, Coimbatore, Tamilnadu.
Phytochemical Screening Test
The phytochemical screening of the plant extract was carried out by following methods used by
Amarasingham et al.,(1964), Das and Bhattacharjee, (1970), Gibbs, (1974), Trease and Evans (1978), Santaram and
Harborne (1984) to detect the presence or absence of certain bioactive compounds:
Anti bacterial assay
Three different concentrations of the leaf extracts were tested for antibacterial activity using agar disc
diffusion assay according to the method of Bauer et al., 1966. The strains obtained were inoculated in conical flask
containing 100 ml of nutrient broth. These conical flasks were incubated at 370C for 24 h and were referred to as
seeded broth. Media were prepared using Muller Hinton Agar (Himedia), dispensed on petri dishes and lawn cultures
were prepared using sterile cotton swabs with the test organisms from the seeded broth. Sterile discs of six
millimeter width impregnated with 20 µl of test extract in different concentrations were placed on the upper layer of
the lawn cultures. Streptomycin (10µg/disc) was used as standard.The plates were incubated overnight at 370C.
Antibacterial activity of the plant extract was assayed by measuring the inhibition zone formed around the discs. The
experiment was repeated triplets and the mean values were calculated.
The preliminary phytochemical analysis of the leaf extract revealed the presence of Alkaloids,
Flavonoids, Steroids, Saponins, Terpenoids and Tannins (Table 1).The results obtained from the disc diffusion
assay showed an increasing inhibitory effect on bacterial growth with increasing concentration of the extract.
The extracts showed inhibitory activity on almost all bacterial strains tested. Among all the tested organisms,
the gram negative bacterial strain, K.pneumoniae was found to be more susceptible to the plant extract with an
inhibition zone ranging from 11 mm and the gram positive strains were least susceptible with the inhibition
zone ranging from 10 mm. The antibacterial activity in terms of zone of inhibition was presented in Table 2.
The observed activity may be due to the presence of potent phytochemical constituents in the leaf extracts.
DISCUSSION AND CONCLUSION
Plants have been a veritable source of drugs. However, man tends to ignore the importance of herbal
medicine. Medicinal plants have formed the basis of health care throughout the world since the earliest days of
humanity and are still widely used and have considerable importance in international trade (Patrick Ekong Ebong, et
al., 2008). The medicinal value of plants lies in some chemical substances that produce a definite physiological
action on the human body. The most important of these bioactive compounds of plants are alkaloids, flavanoids,
tannins and phenolic compounds (Veeramuthu Duraipandiyan et al., 2006).According to World Health Report of
Infectious diseases 2000; overcoming antibiotic resistance is the major issue of the WHO for the next millennium.
Hence the last decade witnessed an increase in the investigation of plants as a source of human disease management.
Antibacterial property of Ethanol and Methanol leaf extracts of Spathodea campanulata was already proved
against standard strains of Klebsiella pneumonia (Parekh and Chanda.S, 2007). The same result was proved with
petroleum ether and methanol leaf extracts of Spathodea campanulata in our study. The antibacterial activity of
petroleum ether leaf extracts showed better result and the strains used in the study were purely isolated from hospital
environment. Further studies are essential for the isolation of the therapeutic antimicrobials and carry out
pharmacological evaluation of Spathodea campanulata.
• Amarasingam, R.P., Bisset, N.G., Millard, A.K. and Woods, M.C., 1964. Phytochemical survey of Malaya,
part III. Alkaloids and Saponins. J. Ecorn.Bot 18, 270-278.
Amusan, O. O. G.; Adesogan, E. K.; Makinde, J. M.,1996. Antimalarial active principles of Spathodea campanulata
stem bark. Phytotherapy Research 10 (8), 692-693.
Amusan, O. O. G.; Msonthi, J. D.; Makhubu, L. P., 1995. Molluscicidal activity of Spathodea campanulata,
Andrachne ovalis, Phytolacca dodecandra and Hypoxis rooperi. Fitoterapia 66, 113-116.
• Asirvatham Doss and Rangasamy Dhanabalan., 2008. Preliminary Phytochemical screening and antibacterial
studies of Leaf extract of Solanum trilobatum Linn. Ethnobotanical Leaflets 12, 638 – 42.
• Bandow JE, Brotz H, Leichert LIO, Labischinski H, Hecker M.,2003 Proteomic approach to understanding
antibiotic action. Antimicro.Agents. Chemotherap. 47, 948-955.
Banerjee, A.; DE, B.,2001. Anthocyanins in some flowers of West Bengal. Journal of Medicinal and Aromatic Plant
Science, Lucknow, v.23, 600-604.
• Bauer A.N, Kirby, W.M.M. Sherries, J.C and Truck M., 1966 Antibiotic susceptibility testing by a
standardized single disk method. Amer.J.Clin.Pathol 45,493-496.
• Colombo ML, Bosisio E., 1996. Pharmacological activities ofChelidonium majus L (Papaveraceae).
Pharmacol. Res 33, 127-134.
Das, A.K. and Bhatacharjee, A.K., 1970. A systematic approach to Phytochemical Screening, Trop Sci,1254-58.
Duraipandiyan V, Ayyanar M, Ignacimuthu S., 2006. Antimicrobial activity of some ethnomedicinal plants used by
Paliyar tribe from Tamil Nadu, India. BMC Comp. Alt. Med. 6, 35-41.
El-Hela, A. A., 2001a. Phenolics from Spathodea campanulata Beauv. Leaves. Al-Azhar Journal of Pharmaceutical
Sciences, Cairo, v.27, p.152-162.
El-Hela, A. A., 2001b. A new iridoid glucoside from Spathodea campanulata Beauv. leaves. Al-Azhar Journal of
Pharmaceutical Sciences, Cairo, v.27, p.115-120.
El-Seedi HR, Ohara T, Sata N, Nishiyama S.,2002. Antimicrobial terpenoids from Eupatorium glutinosum
(Asteraceae). J.Ethnopharmacol 81, 293-296.
Gibbs, R.D., 1974. Chemotaxonomy of flowering plants. Vol. 1McGill-Queens University Press- N, Montreal-
• Jardim, M. A.; Mendonça, L. F. R.; Ferreira, M.M. D. Os produtos naturais para odesenvolvimento
http://www.mdic.gov.br/tecnologia/revistas/artigos/200112pa/mariojardim.pdf.>. Acesso em: 20 dez. 2003.
• Joly, A. B. Introdução à taxonomia vegetal. 7. ed. São Paulo: Ed. Nacional, 1985. 503p.
Makinde, J. M.; Amusan, O. O. G.; Adesogan, E. K., 1988. The antimalarial activity of Spathodea campanulata
stems bark extract on Plasmodium berghei berghei in mice. Planta Medica, Stuttgart 54(2), 122-125.
Mendes, N. M.; Souza, C. P.; Araújo, N.; Pereira,J. P.; Katz, N., 1986.Atividade moluscicida de alguns produtos
naturais sobre Biomphalaria glabrata. Memórias do Instituto Oswaldo Cruz, Rio de Janeiro, v.81, p.87-91.
Ngouela, S.; Nyasse, B.; Tsamo, E.; Sondengam, B. L.; Connolly, J. D., 1990.Spathodic acid: a triterpene acid from
the stem bark of Spathodea campanulata. Phytochemistry, Oxford, v.29, n.12, p.3959-3961.
Ngouela, S.; Tsamo, E.; Sondengam, B. L., 1988.Extractives from Bignoniaceae: constituents of the stem bark of
Spathodea campanulata. Planta Medica, Stuttgart 54(5), 476.
Ngouela, S.; Tsamo, E.; Sondengam. B. L.; Connolly, J. D. Spathodol, a new polyhydroxysterol from the leaves of
Spathodea campanulata. Journal of Natural Products, Washington, v.54, n.3, p.873-876, may 1991.
Niyonzima, G.; Laekeman, G.; Witvrouw, M.; Van Poel, B.; Pieters, L.; Paper, D.; Clercq, E.; Franz, G.; Vlietinck,
A. J., 1999.Hypoglycemic, anticomplement and anti-HIV activities of Spathodea campanulata stem bark.
Phytomedicine, Jena 6 (1), 45-49.
Parekh J and Chanda S., 2007. In vitro screening of antibacterial activity of aqueous and alcoholic extracts of
various Indian plant species against selected pathogens from Enterobacteriaceae. African Journal of Microbiology
Research Vol. 1 (6), 092-099.
Parekh J, Chanda S., 2007a. In vitro antimicrobial activity and phytochemical analysis of some Indian medicinal
plants. Turk. J. Biol 31, 53-58.
• Patrick Ekong Ebong, Item Justin Atangwho, Eyong Ubana Eyong and Godwin Eneji Egbung,The
Antidiabetic Efficacy of Combined Extracts from Two Continental Plants: Azadirachta indica (A. Juss)
(Neem) and Vernonia amygdalina (Del.)., 2008. (African Bitter Leaf) American Journal of Biochemistry and
Biotechnology 4 (3), 239-244.
• Phytochemical, UNESCO, 1996. Culture and Health, Orientation Texts –World Decade for Cultural
Development 1988 – 1997, Document CLT/DEC/PRO – 1996, Paris, France, pgs. 129.
• Pianaro, A; Dalva Trevisan Ferreira; Jurandir Pereira Pinto; Noemia Kazue Ishikawa; Raimundo Braz -
Filho.,2007. Iridoid glucoside and antifungal phenolic compounds from Spathodea campanulata roots.
Semina: Ciências Agrárias, Londrina 28(2), 251-256.
• Rangasamy Dhanabalan, Asirvatham Doss, Muthusamy Jagadeeswari, Rajamanickam Karthic, Muthusamy
Palaniswamy and Jeyaraman Angayarkanni., 2008. Preliminary Phytochemical Screening and Antimalarial Studies
of Spathodea campanulatum P. Beauv Leaf Extracts. Ethnobotanical Leaflets 12, 811-19.