ANTIBACTERIAL ACTIVITY AND COMPARATIVE...
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ABSTRACTS : The success of chemotherapy lies in the continuous search for new drug to counter the challenge posted by resistant strains. Lantana camara is known to be important plant and it is used for medicinal purpose by local tribal people in tropical regions where this plant is widely available. The medicinal plant Lantana camara was screened for the phytochemical components and its antibacterial activity against common pathogenic organism such as Escherichia coli, Staphylococcus aureus , Proteus vulgaris, Bacillus subtilis, Salmonella typhi. Aqueous extract of leaves and flower of plant species Lantana camara used in indian folklore medicine for the treatment of various bacterial infection were investigated. The phytochemical analysis of aqueus extact of leaves , pink and red flower revealed the presence of alkaloid, anthraquinone, tannins, saponins, phenolics, coumarins, steroids etc. Antibacterial activity was studied using agar well diffusion technique. All the extract showed positive activity against common pathogenic bacteria such as Escherichia coli, Staphylococcus aureus , Proteus vulgaris, Bacillus subtilis and Salmonella typhi. Keyword : Antibacterial activity, Lantana camara, Phytochemical analysis , Aqueous extract INTRODUCTION: MATERIALS AND METHODS: ANTIBACTERIAL ACTIVITY AND COMPARATIVE PHYTOCHEMICAL ANALYSIS OF FLOWERS AND LEAVES OF LANTANA CAMARA VIJAY S.WADHAI Department of Microbiology, Sardar Patel Mahavidyalaya, Chandrapur, 442402, Maharashtra, India. Corresponding author email : [email protected] BIONANO FRONTIER VOL. 5 (2 - II) NOV. 2012 ISSN 0974-0678 315 Plants have been used in traditional medicine for several thousand years. Lantana camara Linn. (Verbenaceae) is a hardy, evergreen, straggling shrub with characteristic odour, it grows up to 3 m height, with or without minute prickles on the [1] branches. It is among top ten invasive weeds on the earth . Lantana camara is known for the presence of many biologically active compounds. All parts of this plant have been used traditionally for several ailments throughout the world. The leaves of this plant were used as an antitumeral, [2] antibacterial, and antihypertensive agent , roots for the [3] treatment of malaria, rheumatism, and skin rashes . Several tri- terpenoids, flavonoids, alkaloids, and glycosides isolated from this plant are known to exert diverse biological activities . Extract from the leaves of L. camara possessed larvicidal activity while extract from flowers of the plant showed [4] repellent activity against mosquitoes . Traditionally Lantana camara is considered antiseptic, antispasmodic, carminative and diphoretic. Antiinfflamatory, antipyretic and analgesic properties of extracts of Lantana camara leaves has been reported. Lantana camara is used in herbal medicine for the treatment of skin itches, as an antiseptic for wounds, and externally for leprosy and scabies. Major natural products investigated in Lantana camara belong to the group of triterpinoides, flavonoides and other compounds. In herbal medicine, infusions of the leaves and other plant parts are used as an anti-inflammatory , a tonic and expectorant, and added to baths as an antirhumatic. Lantana [5] extracts have also been shown to be a powerful febrifuge . Because the leaves and some other parts of lantana are poisonous, care must be taken when it is used medicinally. The ripe fruit is benign and heavily consumed by birds and [6] frequently eaten by humans in some countries . Lantana camara is known to have activity against many disease causing organisms. Many research studies carried out till date shows that this weed is positively active against many bacteria, fungi as well as nematodes. The present study is aimed to study the phytochemicals present in L. camara plants bearing pink ,yellow flowers and leaves. In addition, their antibacterial activities have also been studied against common pathogenic organisms. The study also focused on medicinal use of the plant and to bring out the potential of this plant in front of scientific community to explore its medicinal value. Plant Material and Extract Preparation : The leaves and flowers of L.camara were collected locally from old government engineering college area, Chandrapur and identified by the plant taxonomist. The pink and yellow color flowers of L.camara were separated from each single bunch and processed separately. Materials were cleaned properly and allowed to dry for 7 to 8 days under shade. Shade dried materials were dried again using oven at temperature less than o 45 C for 2 to 3 hrs for preventing any kind of contamination during rainy season. Dried material was powdered using pestle and mortar. These powdered material were stored in air tight plastic bags for further use. 15 g of coarsely powdered pink and yellow flowers and leaves of Lantana camara were added to 100 ml double distilled water separately , 2 ml of formaldehyde was added and kept for 2 days at room temperature for cold maceration process. The bottle were shaken for proper extraction. After completion of extraction the mixture was filtered carefully by cotton wool. After filtration half of the solvent was evaporated in hot water bath in order to make it concentrated. The dry weight of pink ,yellow flower and leaves extract were taken. The extract was subjected to preliminary phytochemical tests. Phytochemical Screening method : Preliminary phytochemical screening of plant was done according to the [7],[8],[9],[10] standard procedures adopted by the various workers Test for saponins: Saponins were tested in water extracts. 2 ml of the extract was shaken vigorously for 10 seconds and allowed to stand . The formation of a persistent honeycomb like froth is the positive test for the presence of saponins. Test for tannins: To an aliquot of the extract (dissolved in water) 2 mL of sodium chloride (2%) was added, filtered and mixed with 5 mL 1% gelatin solution. Precipitation indicates the presence of tannins. Test for Triterpenes: Extract (300 mg) was mixed with 5 ml chloroform and warmed for 30 minutes. Few drops of concentrated sulphuric acid was added and mixed well. The appearance of red color indicates the presence of triterpenes.
Transcript of ANTIBACTERIAL ACTIVITY AND COMPARATIVE...
ABSTRACTS : The success of chemotherapy lies in the continuous search for new drug to counter the challenge posted by resistant strains. Lantana camara is known to be important plant and it is used for medicinal purpose by local tribal people in tropical regions where this plant is widely available. The medicinal plant Lantana camara was screened for the phytochemical components and its antibacterial activity against common pathogenic organism such as Escherichia coli, Staphylococcus aureus , Proteus vulgaris, Bacillus subtilis, Salmonella typhi. Aqueous extract of leaves and flower of plant species Lantana camara used in indian folklore medicine for the treatment of various bacterial infection were investigated. The phytochemical analysis of aqueus extact of leaves , pink and red flower revealed the presence of alkaloid, anthraquinone, tannins, saponins, phenolics, coumarins, steroids etc. Antibacterial activity was studied using agar well diffusion technique. All the extract showed positive activity against common pathogenic bacteria such as Escherichia coli, Staphylococcus aureus , Proteus vulgaris, Bacillus subtilis and Salmonella typhi.
Keyword : Antibacterial activity, Lantana camara, Phytochemical analysis , Aqueous extract
INTRODUCTION:
MATERIALS AND METHODS:
ANTIBACTERIAL ACTIVITY AND COMPARATIVE PHYTOCHEMICAL ANALYSIS OF FLOWERS AND LEAVES OF LANTANA CAMARA
VIJAY S.WADHAIDepartment of Microbiology, Sardar Patel Mahavidyalaya, Chandrapur, 442402, Maharashtra, India.
Corresponding author email : [email protected]
BIONANO FRONTIER VOL. 5 (2 - II) NOV. 2012ISSN 0974-0678315
Plants have been used in traditional medicine for several thousand years. Lantana camara Linn. (Verbenaceae) is a hardy, evergreen, straggling shrub with characteristic odour, it grows up to 3 m height, with or without minute prickles on the
[1]branches. It is among top ten invasive weeds on the earth . Lantana camara is known for the presence of many biologically active compounds. All parts of this plant have been used traditionally for several ailments throughout the world. The leaves of this plant were used as an antitumeral,
[2]antibacterial, and antihypertensive agent , roots for the [3]treatment of malaria, rheumatism, and skin rashes . Several
tri- terpenoids, flavonoids, alkaloids, and glycosides isolated from this plant are known to exert diverse biological activities . Extract from the leaves of L. camara possessed larvicidal activity while extract from flowers of the plant showed
[4]repellent activity against mosquitoes .Traditionally Lantana camara is considered antiseptic, antispasmodic, carminative and diphoretic. Antiinfflamatory, antipyretic and analgesic properties of extracts of Lantana camara leaves has been reported. Lantana camara is used in herbal medicine for the treatment of skin itches, as an antiseptic for wounds, and externally for leprosy and scabies. Major natural products investigated in Lantana camara belong to the group of triterpinoides, flavonoides and other compounds. In herbal medicine, infusions of the leaves and other plant parts are used as an anti-inflammatory , a tonic and expectorant, and added to baths as an antirhumatic. Lantana
[5] extracts have also been shown to be a powerful febrifuge . Because the leaves and some other parts of lantana are poisonous, care must be taken when it is used medicinally. The ripe fruit is benign and heavily consumed by birds and
[6]frequently eaten by humans in some countries .Lantana camara is known to have activity against many disease causing organisms. Many research studies carried out till date shows that this weed is positively active against many bacteria, fungi as well as nematodes. The present study is aimed to study the phytochemicals present in L. camara plants bearing pink ,yellow flowers and leaves. In addition, their antibacterial activities have also been studied against common pathogenic organisms. The study also
focused on medicinal use of the plant and to bring out the potential of this plant in front of scientific community to explore its medicinal value.
Plant Material and Extract Preparation : The leaves and flowers of L.camara were collected locally from old government engineering college area, Chandrapur and identified by the plant taxonomist. The pink and yellow color flowers of L.camara were separated from each single bunch and processed separately. Materials were cleaned properly and allowed to dry for 7 to 8 days under shade. Shade dried materials were dried again using oven at temperature less than
o45 C for 2 to 3 hrs for preventing any kind of contamination during rainy season. Dried material was powdered using pestle and mortar. These powdered material were stored in air tight plastic bags for further use.15 g of coarsely powdered pink and yellow flowers and leaves of Lantana camara were added to 100 ml double distilled water separately , 2 ml of formaldehyde was added and kept for 2 days at room temperature for cold maceration process. The bottle were shaken for proper extraction. After completion of extraction the mixture was filtered carefully by cotton wool. After filtration half of the solvent was evaporated in hot water bath in order to make it concentrated. The dry weight of pink ,yellow flower and leaves extract were taken. The extract was subjected to preliminary phytochemical tests.Phytochemical Screening method : Preliminary phytochemical screening of plant was done according to the
[7],[8],[9],[10]standard procedures adopted by the various workers Test for saponins: Saponins were tested in water extracts. 2 ml of the extract was shaken vigorously for 10 seconds and allowed to stand . The formation of a persistent honeycomb like froth is the positive test for the presence of saponins. Test for tannins: To an aliquot of the extract (dissolved in water) 2 mL of sodium chloride (2%) was added, filtered and mixed with 5 mL 1% gelatin solution. Precipitation indicates the presence of tannins. Test for Triterpenes: Extract (300 mg) was mixed with 5 ml chloroform and warmed for 30 minutes. Few drops of concentrated sulphuric acid was added and mixed well. The appearance of red color indicates the presence of triterpenes.
BIONANO FRONTIER VOL. 5 (2 - II) NOV. 2012ISSN 0974-0678316
Test for alkaloids: Extract (300 mg) was digested with 2 M HCl, and the acidic filtrate was mixed with amyl alcohol at room temperature. Pink colour of the alcoholic layer indicates the presence of alkaloids. Test for flavonoids: The presence of flavonoids was determined by using 1% aluminum chloride solution in methanol, concentrated HCl, magnesium and potassium hydroxide solution. Screening for Antimicrobial activity : Agar well diffusion
[11]method was employed .Antibacterial activity of flowers and leaves of Lantana camara plant studied with aqueous extract obtained by cold maceration process. A sterile cotton swab was dipped into bacterial suspension and then was evenly spreaded over the entire surface of nutrient agar plate to obtain uniform inoculums. Escherichia coli ,Staphylococcus aureus, Salmonella typhi, Proteus vulgaris and Bacillus subtilis were the organisms tested. A well was punched on each seeded plate using a sterile borer of 8 mm and the plates were allowed to dry for 5 min. 50 µl of aqueous extracts were dispensed into each well. The
oplates were incubated overnight for 24 hrs at 37 C. Antibacterial activity was determined by measuring the diameter of zone of inhibition in mm with the help of mm scale.
The qualitative analysis of the extracts of leaf and flower sample of Lantana camara showed the presence of alkaloids, anthocyanins, anthocyanidins, anthraquinones, saponinn, tannin and triterpenoids ( Table 1)Table 1: Phytochemical analysis of Leaves and flower of L.camara
RESULT AND DISCUSSION :
the table shows that all extract shows positive activity against two gram positive and three gram negative bacteria. Aqueous extract of flower showed highest activity against S. aureus i.e.30 mm(dia. Of zone of inhibition) whereas aqueous extract of leaves showed highest activity against Bacillus subtilis i.e. 22 mm and Salmonella typhi i.e. 24 mm. Aquous extract of flower and leaves extract showed highest activity against P.vulgaris i.e. 28 mm. Antibacterial activities of L. camara leaf and flower extract might be due to presence of some of these chemical constituents particularly presence of phenolics, anthocyanins, alkaloids, triterpenoids could be responsible for antibacterial properties
1. Sharma G.P., Raghubanshi A.S. and Singh J.S., 2005, Lantana invasion: An overview. Weed Biol. Manage.; 5:157165.
2. Taoubi K, Fauvel M.T, Gley J and Moulis C, 1997, Phenylepropanoid glycosides from Lantana camara and Lippia multoflora. Planta Med.;63:192-3.
3. Chharba S.C., Mahunnah R.L.A. and Mshiu E.N.,1993, Plants used in traditional medicine in eastern Tanzania. J Ethnopharmacol.; 39:83-103.
4. Dua V.K, Pandey A.C., Singh R, Sharma V.P. and Subbarao S.K.,2003,Isolation of repellent ingredients from Lantana camara (Verbenaceae) flowers and their repellency against Aedes mosquitoes. J Appl Entmol ; 12 :509-11.
5. Liogier H.A.1990. Plantas medicinales de Puerto Rico y del Caribe. Iberoamericana de Ediciones,Inc., San Juan, 1990; 563.
6. Herzog, Farah F.Z, and Amado R. 1994, Composition and consumption of gathered wild fruits in the V-Baoule, Cote d'Ivoire. Ecology of Food and Nutrition ; 32(3-4):181-196.
7. Danial K.,Biochemical constituents of biological importance; Indiana press publication; volume4;1991;224-226
8. H a r b o u r n e L . , G u i d e f o r p h y t o c h e m i c a l analysis;edition1.1984.rev.edi 1998, 54-55
9. David P.J., Pharmacology: secondary metabolites sp.edit;Kevin publication.2000, 433-435
10. Kokate G.L.,Skirsagar K,A.,Walke W.N., Phytochemistry of secondary metabolites, vol.3 2004,167-169
11. Deena M. J.,Thoppil J.E. (2000), Antimicrobial activity of essential oil of Lantana camara:Fitoterapia, 71:453-455
12. Kong C.H.(2006), Weed research, volume 46,issue 4 ,290-29513. Ganjewala D.,Sam S.,Khan K.H.(2009),Biochemical
composition and antibacterial activities of Lantana camara plants with yellow,lavender,red and white flowers, Eur asian journal of Biosciences;72
REFERENCES :
Table 2: Antibacterial activities of L. camara leaf and flower extract
+ = Present , - = Absent
Values are mean of triplicate determination on dry weight basis
Similar phytochemical analysis was performed showed the presence of pentacyclic triterpenoids in leaf extracts of
[12]Lantana camara . In literature, it has been also documented the presence of tannins and triterpenoids in aerial parts of Lantana camara, presence of alkaloids , glycosides, tannins in
[13]leaves . This shows flowers and leaves of Lantana camara has reservoir of secondary metabolites. Further technique can be used to characterize these secondary metabolites. These secondary metabolite are known to be responsible for antimicrobial and antioxidant activities.Antibacterial activities of L. camara leaf and flower extract are presented in Table.2
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