Evaluation of preliminary phytochemical and antibacterial properties of Euphorbia heterophylla L. ethanolic extract collected from the selected villages of Tumkur
Chidanandamurthy G1, Ashwathanarayana R2.
Corresponding Author: jamadagni341@gmail.com
ABSTRACT
Medicinal plants have been identified and used throughout human history. We aimed to evaluate phytochemical and antibacterial properties of Euphorbia heterophylla ethanolic extract. This experiment was conducted using standard procedures mainly focused on preliminary phytochemical analysis and antibacterial activity. The results revealed that ethanolic extract give positive result for alkaloids, saponins, flavonoids, steroids, glycosides, phenols and sterols. The antibacterial analysis revealed that the ethanolic extract of E. heterophylla showed maximum zone of inhibition against Pseudomonas aeruginosa followed by Staphylococcus aureus, Xanthomonas campestris, Salmonella typhi, Klebsiella pneumoniae Pseudomonas syringae and least was Escherichia coli. The ethanolic extract of E. heterophylla has potential antibacterial agents which further investigation needed.
KEYWORDS : Antibacterial activity, Euphorbia heterophylla, ethanolic extract, preliminary phytochemical analysis.
INTRODUCTION
Plants make many chemical compounds for biological functions, including defense against insects, fungi and herbivorous mammals.
Antimicrobial chemical is an agent that kills microorganisms or inhibits their growth. The discovery, development and clinical use of antibacterial during the 20th century. Antimicrobial chemicals medicines can be grouped according to the microorganisms hey act primarily against. For example, antibiotics are used against bacteria and antifungals are used against fungi. Antibacterial chemicals are used to treat bacterial infections. The toxicity to humans and other animals from antibacterial is generally considered low.
The aim of the research topic is to evaluate the preliminary phytochemical and antibacterial properties of E. heterophylla collected from the selected villagers of Tumkur.
E. heterophylla is widely used in traditional African medicine and elsewhere in tropical countries. In Africa, a decoction or infusion of the stems and fresh or dried leaves is taken as a purgative and laxative to treat stomachache and constipation, and to expel intestinal worms. A leaf infusion is used as a wash to treat skin problems, including fungal diseases, and abscesses (Abubakar et al., 2013).
In Nigeria, the latex and preparations of the leaves and root are applied to treat skin tumours. In East Africa, the roots are used in the treatment of gonorrhoea or to increase milk production in breast-feeding women. The latex is irritant to the skin and eyes and may be employed as a rubefacient and to remove warts. However, the latex is also used as an antidote against the irritation caused by the latex of other Euphorbia species. In peninsular Malaysia, a leaf extract is taken to treat body pain. The latex is used in the preparation of arrow poison and fish poison (Aniel et al, 2010)
In Bengal the leaves are eaten as a vegetable or famine food, despite their laxative action. The plant is grazed by livestock, and can be fed to guinea pigs as an addition to fresh forage. Honeybees collect the nectar from the flowers (Asha et al, 2014).
All parts of E. heterophylla contain latex: leaves 0.42%, stems 0.11%, roots 0.06% and whole plant up to 0.77%. In Sudan, it is reported to have been exploited during World War II as a substitute for rubber, but this has never been followed up. The plant furthermore contains lectins and carbohydrates. A dimeric N-acetylgalactosamine-specific lectin was isolated from the seeds. Germinating seeds contain endo-1,4-β-glucanases which degrade carboxymethylcellulose (Chinwe et al., 2012; Emele et al., 1998; Perumal et al., 2006).
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Figure 1: Habit of Euphorbia heterophylla |
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MATERIALS AND METHODS
Plant collection and authentication
The plant materials were collected in the villages around Tumkur city, Karnataka in January 2019.(13.367190° N, 77.101176° E) The plants were identified by Dr.Ashwathanarayana R, Guest Faculty, SSCASCW, Tumkur and voucher specimen was conserved under the reference number SSCASCW/DB/GC/001-003 (Figure 2).
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Figure 2: location where plants were collected |
Plant preparation and extraction
The plant samples were dried in shade for 20 to 25 days, mechanically powdered and were collected in airtight plastic containers and stored in dry condition.
Preparation of extracts
20 g of shady dried powder of the selected plants was soaked in ethanol for 30 days, filtered the soaked extract, and preserved for further tests.
Phytochemical Analysis
Preliminary phytochemical analysis done through standard procedures (Harborne, 1998).
Antimicrobial activity
Pathogenic bacterial strains like Xanthomonas campestris (MTCC-2286), Pseudomonas syringae (MTCC-1604), Agrobacterium tumefaciens (MTCC-431), Klebsiella pneumonia (MTCC-7028), Escherichia coli (MTCC 1559), Salmonella typhi (MTCC-734), Pseudomonas aeruginosa (MTCC-1934), Staphylococcus aureus (MTCC-902). Streptomyces pneumonieae (MTCC-4734) obtained from the Institution of Microbial Technology (IMTECH), Chandigarh, India were used. Nutrient agar (NA) media used for the culturing of experimental fungal pathogens.
The test pathogenic bacteria were aseptically inoculated in sterile test tubes using nutrient broth and incubated at 37ºC for 24 hours. The plant crude extracts were dissolved in 10% DMSO to get desired concentrations of 12.5, 25, 50 and 100 mg/ml respectively. The drug Ciproflaxin was used as a standard antibiotic (1mg/ml of sterile distilled water) to compare with the plant crude extracts. Nutrient agar plates were prepared and the broth cultures of bacterial strains were uniformly swabbed. 0.6 cm diameter wells were punched in the inoculated plates using a sterile cork borer. 100 µl of different concentrations of crude extracts and standard (Ciproflaxin, 1mg/ml of sterile distilled water) and DMSO (10%) were filled into the respectively labeled wells and incubate for 24 hours at 37ºC (Aref, 2010).
RESULTS
Preliminary qualitative phytochemical analysis of E. heterophylla leaf ethanolic extract
Cold extraction of E. heterophylla leaf (700 grams) with ethanol 28.69 grams of yield. The preliminary phytochemical analysis of E. heterophylla leaf ethanolic extract were given in Table 1. The preliminary phytochemical analysis of ethanolic extract give positive result for alkaloids, saponins, flavonoids, steroids, glycosides, phenols and sterols.
Table 1: Preliminary qualitative phytochemical analysis of E. heterophylla leaf ethanolic extract
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Sl. No. |
Secondary Metabolites |
Name of the Test |
Ethanolic Extract |
|
1 |
Alkaloids |
Mayer’s test |
+ |
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Wagner’s test |
+ |
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2 |
Saponins |
Foam test |
+ |
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3 |
Tannins |
Ferric chloride test |
- |
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Ferric chloride test |
+ |
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Shenoda test |
+ |
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4 |
Flavonoids |
Zinc HCl reduction test |
- |
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Alkaline reagent test |
+ |
||
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Lead acetate test |
+ |
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5 |
Steroids |
Salkowski test |
+ |
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Keller-Kiliani test |
+ |
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6 |
Glycosides |
Legal's test |
- |
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Ferric chloride test |
+ |
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7 |
Phenols |
Ellagic acid test |
- |
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8 |
Sterols |
Liebermann Burchard test |
+ |
Antibacterial activity of E. heterophylla
Ethanolic extract of E. heterophylla leaf showed appreciable antibacterial activity in all tested concentrations (12.50, 25, 50, 100 mg/ml) which is showed in the Table 2. In 100 mg/ml concentration maximum zone of inhibition showed by Pseudomonas aeruginosa (23.33±0.66) followed by Staphylococcus aureus (22±1.15), Xanthomonas campestris (21.33±0.88), Salmonella typhi (21±1.54), Klebsiella pneumoniae (20.33±1.2), Pseudomonas syringae (17±0.57) and least was Escherichia coli (16±0.57) control DMSO shows nil zone and standard ciprofloxacin showed maximum zone of inhibition for all the pathogenic bacteria (Figure 3).
Table 2: Antibacterial activity of E. heterophylla ethanolic extract
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Ethanolic extract |
Zone of Inhibition in mm (Mean±SEM) |
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Test Organisms |
12.50 mg/ml |
25 mg/ml |
50 mg/ ml |
100 mg/ml |
Control |
Standard |
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Xc |
9.66±0.88 |
14±1.15 |
14.66±0.88 |
21.33±0.88 |
0±0 |
30.66±0.66 |
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Ps |
9±1.15 |
12.33±1.45 |
15.33±1.45 |
17±0.57 |
0±0 |
28.66±0.66 |
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Kp |
9.33±1.45 |
10.33±0.88 |
13.66±0.88 |
20.33±1.2 |
0±0 |
30.33±0.33 |
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Ec |
0±0 |
11.33±1.2 |
12±1.15 |
16±0.57 |
0±0 |
32±0.57 |
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St |
10.33±1.45 |
17.33±0.88 |
19.33±1.45 |
21±1.54 |
0±0 |
32.33±0.33 |
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Pa |
9.33±88 |
19.33±0.88 |
21.33±0.88 |
23.33±0.66 |
0±0 |
31±0.57 |
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Sa |
10.33±0.88 |
16.66±0.88 |
21±0.57 |
22±1.15 |
0±0 |
33.66±0.33 |
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Xc- Xanthomonas campestris, Ps- Pseudomonas syringae, Kp- Klebsiella pneumoniae, Ec- Escherichia coli, St- Salmonella typhi,Ps- Pseudomonas aeruginosa, Sa-Staphylococcus aureus
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Figure 3: Antibacterial activity of Euphorbia heterophylla ethanolic leaf extract |
DISCUSSION
Preliminary phytochemical analysis
The ethanolic solvent yields appreciable amount of extract for E. heterophylla leaf sample. The preliminary phytochemical analysis of E. heterophylla leaf ethanolic extract also revealed the presence of excellent phytoconstituent like alkaloids, saponins, flavonoids, steroids, glycosides, phenols and sterols.
Antimicrobial properties
The ethanolic extracts of E. heterophylla leaf showed appreciable antibacterial activity in all tested concentrations (12.50, 25, 50, 100 mg/ml) which is due to the phytochemical dissolved in it and antibacterial compounds present in it. In 100 mg/ml concentration maximum zone of inhibition showed by Pseudomonas aeruginosa followed by Staphylococcus aureus, Xanthomonas campestris, Salmonella typhi, Klebsiella pneumoniae Pseudomonas syringae and least was Escherichia coli. All the tested pathogens were belonging to plant and animal pathogens, some of them were opportunistic pathogens. In recent years drug resistant bacterial strains were dangerous threat to human beings and new antibacterial drug synthesis is necessary to get rid these drug resistance bacterial strains so, drug which has multiple pathogenic targets and safer to consume is appreciable in that concern plant origin compounds were safety as well as cost effective. Control DMSO shows nil zone in turn confirm the positive effect of ethanolic extract.
CONCLUSION
The presence of saponins and alkaloids has been reported to be responsible for various pharmological properties with exerting toxic effects against cells of foreign organisms. The mehanolic extract of E. heterophylla could be attributed to the presence of the alkaloids, cardiac glycosides, tannins, saponins etc. in the whole plant of selected plants therefore; this shows that the plant could be useful as antitumor, anticancer and as antimicrobial agents. The present study revealed the best phytochemicals a well as primary and secondary metabolites in the selected plants it may useful information for biochemical reactions and biological activities in the selected plants. By the rich source of the phyto-constituents in the selected plant, these plants were used in the treatment of respiratory, urinary tract and gastrointestinal infections in traditional medical practice is justified.
REFERANCES
