Phytochemical, Antimicrobial and Antioxidant

Muhammad Ajaib et al.,

J.Chem.Soc.Pak., Vol. 38, No. 03, 2016

538

Phytochemical, Antimicrobial and Antioxidant Screening of Fruits, Bark and leaves of Lagerstroemia indica 1

Muhammad Ajaib*, 2Taha Arooj, 3Khalid Mohammed Khan**,4Sidra Farid, 1 Muhammad Ishtiaq, 5Shahnaz Perveen and 3Shazia Shah 1 Department of Botany, Mirpur University of Science & Technology, Bhimber Campus, AJK, Pakistan 2 Department of Botany, GC University Lahore Pakistan. 3 H. E. J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences,University of Karachi, Karachi-75270, Pakistan. 4 Department of Chemistry, GC University Lahore Pakistan. 5 PCSIR Laboratories Complex, Shahrah-e-Dr. Salimuzzaman Siddiqui, Karachi-75280, Pakistan. [email protected]*, [email protected]** (Received on 31st August 2015, accepted in revised form 20th January 2016) Summary:The present study was conducted to evaluate phytochemicals, antimicrobial and antioxidant potential of Lagerstroemia indica L. The phytochemical screening of L. indica revealed the presence of active metabolites such as anthraquinones, reducing sugars, terpenoids, flavonoids, saponins, tannins, alkaloids and cardiac glycosides. Antimicrobial assessment was carried out against Gram-positive bacteria (Bacillus subtilis, Staphylococcus aureus), Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli) and fungal strains (Aspergillus oryzae and Aspergillus niger). Maximum antibacterial potential (58.33 ± 0.88 mm) was exhibited by petroleum ether extract of bark against B. subtilis. The maximum antifungal potential 40.33 ± 0.88 mm and 40.0 ± 1.15 mm against A. niger was observed by chloroform extract of bark and fruits respectively. The antioxidant potential was assessed using five assays viz. ABTS activity, DPPH° radical scavenging activity, metal chelating activity, total flavonoid contents and total phenolic contents. Highest TEAC value 7.946 ± 0.04 mM trolox for ABTS+ assay was observed by aquous extract of leaves. The highest values for total flavonoid contents 1185.740 ± 0.01 µg/ml and total phenolic contents 40.333 ± 0.23 µg/ml was exhibited by petroleum ether bark extract. The maximum metal chelating activity 60.302 ± 0.93 was observed by petroleum ether extract of fruit. The highest value of % DPPH° (92.92 ± 0.08 %) was observed by aquous extract of bark.

Key words: Phytochemicals, Lagerstroemia indica L., Antimicrobial, Antioxidant, MIC Introduction Herbal medicine, also known as botanical medicine or phytomedicine refers to using plant seeds, flowers, roots for medicinal purpose. It is very important to improve analysis and quality control along with advances in clinical research to show the value of herbal medicine in treating and preventing disease [1]. Demand of drugs to obtained from plant sources is increasing day by day, which can only be fulfilled by screening medicinal plants with promising biological activity. In developing countries, medicines are quite expensive, so there is a need of investigation of the antimicrobial activity of ethnomedicinal plants [2]. The development of drug resistance in human pathogens against commonly used antibiotics has necessitated a search for new antimicrobial substances from other sources, including plants. Plants used in traditional medicines contain a wide range of substances that are used to treat chronic as well as infectious diseases [3]. Lagerstroemia indica L belongs to family lythraceae. It is commonly known as “Crape myrtle”. It is a perennial shrub and reaches at the height of 10 *

To whom all correspondence should be addressed.

to 30 feet and spreads 10 to 15 feet. It showed many biological activities such as anti- inflammatory, antipyretic, analgesic, antihyperglycemic, antioxidant, hepatoprotective and antihyperglycemical [4]. Previously diterpenoids, triterpenoids, sterols, [5] anthocyanins [6]; lignans [7], alkaloids [8] fatty acids [9] sesquiterpenes and coumarins [10] were reported from different species of Lagerstroemia. The present study was designed to investigate the antimicrobial and antioxidant activities of an ethnobotanically important plant Lagerstroemia indica L. Experimental Chemicals and Reagents All chemicals and reagents were used of the standard grade. Petroleum ether (C2H5OC2H5), chloroform (CHCl3), methanol (CH3OH), distilled water (H2O), ferric chloride, copper sulfate


(CuSO4.5H20), potassium sodium tartrate, sodium hydroxide (NaOH), sulfuric acid (H2SO4), ammonia (NH3), olive oil, acetone, glacial acetic acid, acidic alcohol, mercuric chloride, potassium iodide (KI), bismuth sub-nitrate, conc. hydrochloric acid, ferric chloride, nutrient broth, agar, potato dextrose agar, 2,2-diphenyl-2-picrylhydrazyl (DPPH), ferrozine, ferrous sulfate (FeSO4), sodium carbonate (Na2CO3), ABTS reagent, sodium chloride (NaCl), ferric chloride, sodium acetate, K2HPO4, KH2PO4, sodium nitrate (NaNO2), aluminum chloride (AlCl3) Instruments Electric balance (GE.212 Gottingen, Germany), Oven Memmert (Germany), autoclave, hot plate, laminar air flow hood (model No. PA 12SOO) technico scientific supply, Lahore, Pakistan, incubator (model No.B-53, Sr. No. 1353114, IRMECO, Gecthacht, Germany), spectrophotometer (UV-2300, Tec Comp, Shanghai), rotary evaporator, pH meter Plant Material The plant material Lagerstroemia indica L. was collected from Botanic Garden, GCU Lahore and identified from Dr. Sultan Ahmad Herbarium, Department of Botany, GC University, Lahore with a voucher specimen no. GC. Herb. Bot. 2900. Maceration of Plant Material Leaves, bark and fruit of the plant (500 g) were shade dried and macerated in different solvents (1L) such as petroleum ether, chloroform, methanol, and distilled water respectively. The resultant extracts were dried on rotary evaporator at 40°C to get concentrated extracts. Microorganisms Two gram positive (Staphylococcus aureus and Bacillus subtilis) and two Gram negative (Pseudomonas aeruginosa & Escherichia coli) bacteria were obtained from Punjab Institute of Cardiology, Jail road Lahore. While two fungal strains (Aspergillus oryzae and Aspergillus niger) were obtained from the Institute of Industrial Biotechnology, GC University Lahore. Phytochemical Analysis Qualitative tests for phytochemical screening of plant were performed to assess the presence of active secondary metabolites like


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alkaloids, tannins, reducing sugars, flavonoids, terpenoids, cardiac glycosides, saponins and anthraquinones following the method described by Ayoola et al., [11]. Test for Reducing Sugars (Fehling’s test) The aqueous ethanol extract (0.5 g in 5 ml of water) was added to boiling Fehling’s solution (A and B) in a test tube. The solution was observed for a colour reaction. Formation of yellow to brownish-red precipitates confirmed the presence of reducing sugars. Test for Alkaloids 0.5 g of extract was diluted to 10 ml with acid alcohol, boiled and filtered. To 5 ml of the filtrate, 2 ml of dilute ammonia was added followed by addition of 5 ml of chloroform and shaken gently to extract the alkaloidal base. The chloroform layer was extracted with 10 ml of acetic acid. This was divided into two portions. Mayer’s reagent was added in one portion and Draggendorff’s reagent to the other. The formation of a cream (with Mayer’s reagent) or reddish brown precipitate with (Draggendorff’s reagent) was regarded as positive for the presence of alkaloids. Test for Flavonoids 5 ml of dilute ammonia was added to a portion of an aqueous filtrate of the extract followed by addition of 1ml of concentrated sulfuric acid. A yellow colouration that disappears on standing indicates the presence of flavonoids. Test for Terpenoids (Salkowski test) The test is named after the German biochemist Ernst Leoplod Salkowski and was taken under consideration by Harborne. In each reaction tube, 0.5 g each of the extract was added along with 2 ml of chloroform. Concentrated H2S04 (3 ml) was carefully added to form a layer. A reddish brown colouration of the interface indicates the presence of terpenoids. Test for Cardiac Glycosides (Keller-Killiani test) It was named after C. C. Keller and H. Killiani, whom initially formulated it to examine Digitalis in the 19th century. In this test 0.5 g of extract was diluted to 5 ml in water. 2 ml of glacial acetic acid containing one drop of ferric chloride solution was added to reaction tube. This was


underlaid with 1 ml of concentrated sulfuric acid. A brown ring at the interface indicated the presence of a deoxysugar characteristic of cardenolides. A violet ring may appear below the brown ring, while in the acetic acid layer a greenish ring may form just above the brown ring and gradually spread throughout this layer. Test for Tannins For validation of occurrence of tannins the test was postulated relying postulate that hydrolysis of tannins take place producing Gallic and ellagic acid that are transformed to pyrogalol after filtration and contributes to green to blue hue when reacts with FeCl3. In reaction tube, about 0.5 g of the extract was boiled in 10 ml of water and then filtered. A few drops of 0.1% ferric chloride were added in the filtrate. Detection of brownish green (cathechic tannins) or a blue-black (gallic tannins) colouration confirmed presence of tannins. Test for Saponins (Frothing test) In reaction tube 0.5 g of extract was included in 5 ml of distilled water. The solution was shaken vigorously and observed for a stable persistent froth. The frothing was mixed with 3 drops of olive oil and shaken vigorously. Formation of an emulsion confirmed presence of Saponins. Test for Anthraquinones (Borntrager’s test) The Borntrager’s test was adopted for detection of anthraquinones. 0.5 g of the extract was boiled with 10 ml of sulfuric acid (H2SO4) and filtered while hot. The filtrate was shaken with 5 ml of chloroform that resulted in the separation of distinct organic layers. The chloroform layer was pipette into another test tube and 1 ml of 10% ammonium hydroxide was added. The resulting solution was observed for colour changes. Gradual transition of solution to pink, red or violet colour indicated the occurrence of anthraquinones in plant macerate. Antimicrobial Activity Antimicrobial activity was carried out by using Agar well diffusion method of Jorgenson et al. [5], Cruick-shank et al. [12] and Johansen [13]. Evaluation of Minimum Inhibitory Concentration (MIC) Minimum Inhibitory Concentration (MIC) evaluated by using agar micro dilution method following by Jorgenson et al. [14].


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Agar well Diffusion Method Agar well diffusion technique of Jorgensen et al. was employed in this investigation. According to this technique, dilute inoculums of tested bacteria and fungi were spread on the petri plates containing desired media, standard hole was made in the mid of all petri plates using cork borer no. 4 and 1 ml of crude extract was filled in the hole in the laminar air flow under aseptic conditions. Each plate was labeled, covered with cling film and placed in incubator at 35 ± 2 ○C and 25 ± 2 ○C for 24 h and 48 h investigation of antibacterial and antifungal activity respectively. Measurement of Zone of Inhibition After incubation, diameter of zone inhibited by plant extract was measured with the help of a transparent ruler in mm. In some conditions, asymmetrical zones were obtained. In these cases, the quantification was made from the middle of the spot at the border with distinct boundaries, more than one value of the diameter was noted from dimensions and then their mean value was calculated. Evaluation of Minimum Inhibitory Concentration (MIC) The agar dilution method used by Jorgensen et al. was applied for the investigation of the minimum inhibitory concentration with reference to negative (Solvents: Petroleum ether, Chloroform, methanol and distilled water) and positive (antibiotic dilutions) control. Agar Dilution Method Following agar dilution method, the samesized, pre-labeled petri-plates were autoclaved for 15 minutes at 121○C temperature at 15 psi pressure and mixture comprising of 18 ml of medium (cooled to >50○C) and 2 ml of different concentrations of crude extracts of Lagerstroemia indica L. was added in individual reaction plate, followed by placing a lid on them. These plates were sealed with cling film and placed at room temperature to settle the media. Afterwards, the suspension of microbes (inoculums) was streaked in a zig-zag manner on the prepared petri-plates with the assistance of disinfected cotton mop, the lid of the plate was positioned on it, secured with cling film and incubated at 37 ± 2○ C for 24hrs (bacteria) or at 25 ± 2○ C for 48 hrs (fungi). Measurement of MIC After incubation, petri-plates were analyzed for the presence (+) or absence (-) of microbial


proliferation. The least concentration that had successfully obviated microbial growth was treated as MIC.


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expressed as mg of quercetin equivalents per gram of sample. Total Phenolic Contents

Antioxidant Assay Antioxidant potential of the plant extracts was assessed by ABTS+ assay following Re et al. [15], total flavonoid contents by Dewanto et al., [16], total phenolic contents and metal chelation by Dinis et al., [17] and DPPH° radical scavenging activity by Lee and Shibamoto method [18]. ABTS+ Assay It was initially developed by Miller and Rice and improved by Re et al., For evaluation of ABTS•+ assay 7 mM solution of ABTS was prepared in double distilled water, which generated ABTS•+ while reacting with 2.45 mM potassium persulfate after 24 h of standing under the dark. The absorbance of ABTS•+ stock solution was adjusted to 0.70 ± 0.02 at 734 nm. If the absorbance exceeds this value, it was diluted with PBS buffer of pH 7.4 and if below this value, ABTS reagent was added. For the evaluation of antioxidant activity, add 10 μl of sample to 2.99 ml of diluted solution of ABTS•+ (A=0.70± 0.02) and note the change in absorbance after every 1 min interval for 8 min. Appropriate solvent blank was run in parallel. All the samples were run in triplicate and mean values of absorbance were calculated. A dose response curve of trolox was prepared by plotting its absorbance at 734 nm and % age inhibition for each sample was calculated by using this formula. % age inhibition (at 734nm) = ( 1-Af ) x 100 A0 where A0 is the absorbance of ABTS radical cation and Af is the absorbance after sample addition. Total Flavonoids Contents The TFC content of plant fractions was determined by following method developed by Dewanto et al.,. 0.25 ml of plant sample/quercetin standard solution was mixed with 1250 μl of distilled water in a test tube then 75 μl of NaNO2 solution was added. After 5 min stay, 0.5 mL of 1M NaOH was added and the final volume was raised to 2.5 mL with distilled water.The absorbance of this sample solution was measured at 510 nm. TFC contents were determined from the standard curve of quercetin and

The total phenolic content (TPC) for each plant sample was calculated by the following method. A supersaturated solution of sodium carbonate was prepared. To 40 μl of the sample, add 3.16 ml of double distilled water and 200 μl of Folin-ciocalteu reagent, mix them well and allow it to stay for 8 min. Now, add 600 μl of sodium carbonate solution and allow it to incubate at 40○C for 30 min and note its absorbance at 765 nm. Total phenolic contents were calculated from the standard curve and expressed as mg/g equivalents of gallic acid (GAE). Metal Chelating Activity: The ability of plant extracts to chelate iron (II) was determined by the method devised by Dinis et al.,. The reaction mixture was prepared by mixing 100 μL of sample with 0.05 ml of FeSO4 and 0.2 ml of 5 mM ferrozine. Final volume was raised upto 4 mL with double distilled ethanol. Allow this reaction mixture to stand for 10 min at room temperature and note the absorbance of the solution at 562 nm. The results were expressed as the %age of bound iron, which can be calculated from the formula shown below or in terms of EDTA standard. % age bound iron=(Acontrol –Asample)X Acontrol

100

DPPH Free Radical Scavenging Activity The stock solution of DPPH radical cation was prepared (25 mg/L) in methanol and absorbance was adjusted to 1.00 ± 0.02 at 515 nm by diluting it with methanol. Appropriate quantity of sample was added to 2.5 mL of this diluted reagent and change in absorbance was measured after every 5 min interval for 30 min. The % age of DPPH radical remaining was calculated using the formula. % age DPPH remaining = Af X 100 A0 Kinetic curves were plotted showing a decrease in absorbance of DPPH with time and EC50 value was also calculated for each sample. Statistical Analysis of the Data All the parameters were carried out in triplicates and data was presented as mean value ±


S.E. The mean value, standard deviation and standard error were calculated using Microsoft excel. The results obtained were analyzed statistically by employing analysis of variance (ANOVA) and Duncan’s multiple range tests using co-stat software (version 3.03) to determine the significant value of the analysis. Results and Discussions The phytochemical screening of bark, leaves and fruit extracts of Lagerstroemia indica revealed the presence of alkaloids, tannins, reducing sugars, flavonoids, terpenoids, cardiac glycosides, saponins and anthraquinones in all extracts in different ratios (Fig. 1).


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mm) by petroleum ether bark and minimum zone (11.33 ± 0.33 mm) by petroleum ether fruit extract was observed against E. coli. The highest activity (49.33 ± 0.66 mm) against P. aeruginosa was displayed by petroleum ether leave extract, while least activity (10.33 ± 0.33 mm) was reported by petroleum ether bark extract. Similarly, maximum inhibition (46 ± 2.08 mm) against S. aureus was produced by petroleum ether leaves extract and minimum zone (10.33 ± 0.33 mm) by petroleum ether extract of bark and fruits, as well as, aqueous extract of leaves (Table-2). These results were compared with standard discs. The standard discs, i.e. amikacin, cephalaxin and erythromycin showed less inhibition as compared to plant extracts. Summarizing the results of antibacterial activity, it was observed that chloroform and methanol extracts of bark, leaves and fruits of Lagerstroemia indica were more effective as compared to aqueous and petroleum ether extracts. Table-1: Zone of inhibition (mm) indicated by Standard antibiotic discs against microbial strains. Bacterial Strains E. coli P. aeruginosa S. aureus B. subtilis A. oryzae A. niger

Fig. 1:

Phytochemical profile of different extracts of L. indica.

Antimicrobial study was carried out against two Gram-positive (Staphylococcus aureus and Bacillus subtilis), two Gram-negative (Escherichia coli and Pseudomonas aeruginosa) bacterial strains and two fungal strains (A. oryzae and A. niger). The standard discs were also used in order to make comparison between zones of inhibition produced by commercially available discs and the plant extracts (Table-1). Negative control was employed to provide evidence that antimicrobial potential was observed by plant extracts not by solvents. The maximum antibacterial activity (58.33 ± 0.88 mm) against B. subtilis was shown by petroleum ether bark extract and minimum antibacterial activity (19 ± 0.577 mm) against B. subtilis was exhibited by chloroform leaves extracts The maximum zone (41.33 ± 0.88

Standard disc Amikacin Cephalaxin Erythromycin Amikacin Griesofulvin Terbinafine Griesofulvin Terbinafine

Concentration of Standard discs (µg) 20 20 20 20 20 20 20 20

Zone of Inhibition (mm) 18.7 ± 1.04 18 ± 0.76 15 ± 0.98 17 ± 0.58 17 ± 1.52 15 ± 2.01 18 ± 2.54 15 ± 2.32

The significant antifungal activity was executed by all the extracts of bark, leaves and fruits of Lagerstroemia indica against both fungal strains (Table-3). The largest zone of inhibition (36 ± 3.21 mm) against A. oryzae was exhibited by aqueous extract of bark and minimum zone (19 ±1.15 mm ) by petroleum ether bark extract. Highest antifungal activity (40.33 ± 0.88 mm) was reported by chloroform bark extract against A. niger, while minimum activity 12.66 ± 0.33 mm was displayed by petroleum ether fruits against A. niger. The minimum inhibitory concentration (MIC) was assessed by the agar dilution method verified the antibacterial potential of L. indica and ranges from 5-1.25 mg/ml ( Fig. 2) The MIC of methanolic extracts was evaluated and maximum MIC (1.25 mg/ml) was exhibited by extracts of bark and fruit against B. subtilis and against S. aureus by fruit extract.



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Table-2: Zone of inhibition (mm) produced by L. indica against bacterial strains. Plant parts

Solvents Petroleum ether Chloroform Methanol Distilled water Petroleum ether Chloroform Methanol Distilled water Petroleum ether Chloroform Methanol Distilled water

Bark

Leaves

Fruits

E. coli 41.33 ± 0.88 20.33 ± 0.33 22.33 ± 1.85 16.33 ± 2.18 30.66 ± 0.88 21.0 ± 0.57 17.66 ± 1.45 21.0 ± 0.57 11.33 ± 0.33 30.33 ± 0.33 18.66 ± 0.88 19.33 ± 1.20

LSD

3.275

Zone of inhibition (mm) P. aeruginosa S. aureus 10.33 ± 0.33 10.33 ± 0.33 31.33 ± 0.88 18 ± 1.00 26.66 ± 0.33 18.33 ± 0.88 27.66 ± 0.33 22.33 ± 0.88 46±2.08 49.33 ± 0.66 22.33±0.88 31.0 ± 0.57 23.33 ±1.45 18±1.15 26.66 ± 0.33 10.33±0.33 10.33 ± 0.33 10.66 ± 0.33 31.0 ±1.15 24.66 ± 0.88 26.66 ± 0.33 31.33 ± 0.88 23.33 ± 0.88 18.66 ± 0.88 2.082 2.918

B. subtilis 58.33 ± 0.88 21.66 ± 0.33 38.33 ± 0.88 24.66 ± 0.33 26.66 ± 0.33 19.0 ± 0.57 25.66 ± 0.88 20.0 ± 1.15 19.66 ± 0.33 50.0 ± 0.57 37.0 ± 0.57 31.66 ± 0.33 1.925

Table-3: Zone of inhibition produced by the L. indica extracts against fungal strains. Plant parts

Solvents

Bark

A.oryzae 19.0 ± 1.15 26.0 ± 2.64 22.0 ± 1.53 36.0 ± 3.21 19.33 ± 2.90 25.66 ± 2.18 22.66 ± 1.20 31.0 ± 0.58 19.33 ± 1.20 19.33 ± 3.71 27.66 ±1.45 35.0 ± 2.88 6.617

Petroleum ether Chloroform Methnol Distilled water Petroleum ether Chloroform Methanol Distilled water Petroleum ether Chloroform Methanol Distilled water

Leaves

Fruits

LSD

Zone of inhibition (mm) A. niger 20.33 ± 2.40 40.33 ± 0.88 22.66 ± 1.76 31.0 ± 2.64 20.0 ± 1.15 21.33 ± 0.33 27.33 ± 4.84 20.33 ± 5.48 12.66 ± 0.33 40.0 ± 1.15 19.66 ± 0.88 23.33 ± 1.85 7.409

found by aqueous leaves extract and minimum potential displayed by chloroform extract of fruit. This percentage inhibition activity was compared to trolox used as a standard to determine TEAC values that lie within the range of 1.941 ± 0.24 to 7.946 ± 0.04 mM trolox (Table- 4).

Fig. 2:

Graphical representation of MIC of methanolic extracts of leaves, bark and fruits of L. indica against bacterial strains.

MIC for the antifungal potential of L. indica was observed in the range from 5-1.25 mg/ml. The maximum MIC (1.25 mg/ml) exhibited by methanolic extracts of bark and fruits against A. oryzae and A. niger. Moreover, least potential (5 mg/ml) was concluded by methanolic extracts of leaves and fruits against A. niger (Fig. 3). Antioxidant potential of L. indica was evaluated by using different method. In ABTS+ assay, the % inhibition values of L. indica lied within the range of 22.646 ± 1.68 to 91.316 ± 1.69 % at a concentration of 100 mg/ml with maximum activity

Fig. 3:

Graphical representation of MIC of methanolic extracts of leaves, bark and fruits of L. indica against fungal strains.

The total flavonoids content (TFC) of L. indica was measured by colorimetric method and found within the range of 175.494 ± 0.70 to 1185.740 ± 0.01 µg/mL of quercetin with maximum activity reported by petroleum ether bark extract and minimum efficacy was observed by aqueous extract of fruit (Table- 5).



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Table-4: Total flavonoid contents (expressed in Quercetin equivalent) of L. indica. Plant part

Total Flavonoid Contents Absorbance at 510 nm QUE (µg/ml of Quercetin) 0.734 ± 0.65 266.740 ± 0.09 1.556 ± 1.43 544.662 ± 0.39 2.339 ± 0.99 826.507 ± 0.03 0.548 ± 0.03 191.521 ± 1.35 3.464 ± 0.70 1185.740 ± 0.01 2.448 ± 0.34 845.701 ± 0.48 2.524 ± 0.32 716.581 ± 0.06 0.552 ± 0.08 194.698 ± 0.32 3.236 ± 1.02 1109.526 ± 0.06 2.537 ± 0.48 868.340 ± 0.57 2.528 ± 1.35 729.537 ± 0.99 0.554±0.03d 175.494±0.70l 0.291 0.568


Leaves

Bark

Fruits LSD

Table-5: ABTS+ assay with % inhibition and TEAC estimation of L. indica. Plant part Leaves

Bark

Fruits

Petroleum ether Chloroform Methanol Distilled water Petroleum ether Chloroform Methanol Distilled water Petroleum ether Chloroform Methanol Distilled water LSD

Absorbance at 734 nm 0.914 ± 1.21 0.472 ± 1.30 0.127 ± 1.03 0.065 ± 0.50 0.346 ± 0.50 0.370 ± 0.99 0.135 ± 0.72 0.063 ± 0.85 0.349 ± 1.70 0.562 ± 0.80 0.087 ±1.20 0.063 ±1.68 0.582

The total phenolic contents (TPC) of L. indica was assessed by using Folin-ciocalteu reagent and was found within the range 5.482 ± 0.08 to 40.333 ± 0.23 µg/ml of GAE with the maximum contents reported by petroleum ether bark extract and minimum contents by chloroform extract of leaves (Table- 6). Table-6: Total phenolic equivalent) of L. indica. Plant part

Solvents

Leaves

Petroleum ether Chloroform Methanol Distilled water Petroleum ether Chloroform Methanol Distilled water Petroleum ether Chloroform Methanol Distilled water LSD

Bark

Fruits

ABTS+ Activity % inhibition 63.476 ± 0.49 31.500 ± 0.72 81.830 ± 0.80 91.316 ±1.69 53.536 ± 1.35 47.740 ± 1.20 84.643 ± 0.51 91.064 ± 0.68 53.370 ± 0.99 22.646 ± 1.68 87.570 ± 1.04 90.763±0.99 0.485

Solvents

contents

(Gallic

extract of bark and minimum by aqueous extract of bark (Table-8). Table-7: Metal chelating activity of L. indica. Plant part Leaves

acid

Total Phenolic Contents Absorbance at GAE 765 nm (µg/ml of Gallic acid) 0.394 ± 0.19 6.947±0.85 0.290 ± 1.10 5.482 ± 0.08 0.556 ± 0.84 27.627 ± 1.50 0.843 ± 1.06 14.540 ± 0.06 2.281 ± 0.31 40.333 ± 0.23 0.981 ± 0.50 17.493 ± 0.19 1.228 ± 0.97 22.596 ± 0.90 0.545 ± 0.82 10.551±0.05 1.830 ± 0.97 32.383 ± 0.64 0.316 ± 1.20 5.621 ± 1.20 0.739 ± 1.50 13.502 ± 0.09 0.737 ± 0.69f 12.577 ± 0.01 0.044 0.689

Metal chelating activity of different extracts of L. indica showed percentage of bound iron lies within the range of 0.066 ± 0.63 to 60.302 ± 0.93 % (Table - 7). The % DPPH° radical scavenging activity lies within the range of 32.72 ± 0.56 to 92.92 ± 0.08 % with maximum activity given by methanolic

TEAC (mM of Trolox) 5.711 ± 0.86 2.656 ± 1.04 7.065 ± 0.26 7.946 ± 0.04 4.654 ± 0.49 4.133±1.35 7.331 ± 0.23 7.936 ± 0.27 4.586 ± 1.69 1.941 ± 0.24 7.558 ± 0.51 7.868 ± 0.68 0.131

Bark

Fruits

Metal chelating activity Absorbance at 520nm % bound iron Petroleum ether 1.490 ± 0.09 33.140 ± 0.01 Chloroform 1.675 ± 0.93 8.076±0.63 Methanol 1.357 ± 0.03 24.146 ± 0.54 Distilled water 1.641 ± 0.91 9.436 ± 0.04 Petroleum ether 1.137 ± 0.01 35.838 ± 0.02 Chloroform 1.774 ± 0.02 0.343 ± 0.78 Methanol 1.583 ± 0.71 12.081 ± 0.32 Distilled water 1.635 ± 0.02 9.436 ± 0.03 Petroleum ether 0.736 ± 0.68 60.302 ± 0.93 Chloroform 1.775 ± 0.05 0.066 ± 0.63 Methanol 1.357 ± 0.63 24.170 ± 0.31 Distilled water 1.591 ± 0.01 10.624 ± 0.05 LSD 0.214 0.041 Solvents

Table-8: DPPH° radical scavenging activity of L. indica. Plant part Leaves


Bark

Fruits

LSD

% DPPH° 76.75 ± 0.09 89.59 ± 0.66 77.70 ± 0.72 74.92± 0.32 83.13 ± 0.35 84.96 ± 0.33 32.72 ± 0.56 92.92 ± 0.08 90.89 ± 0.31 76.63 ± 0.13 42.88 ± 0.23 35.27 ± 0.56 0.204


Conclusion The current investigation was an effort to determine the ethnopharmacological characters of Lagerstroemia indica. The antimicrobial potential showed that extracts of L. indica were more effective than standard antimicrobial drugs. Petroleum ether extract of bark showed maximum antibacterial activity 58.33 ± 0.88 mm against B. subtilis. Chloroform extract of bark showed maximum antifungal potential 40.33 ± 0.88 mm against A. niger. Plant extracts also showed significant antioxidant activity. Aqueous extract of leaves showed highest TEAC value 7.946 ± 0.04 mM trolox for ABTS+ assay. Petroleum ether bark extract exhibited maximum value for total flavonoid contents 1185.740 ± 0.01 µg/ml and total phenolic contents 40.333 ± 0.23 µg/ml. Petroleum ether extract of fruit showed maximum metal chelating activity 60.302 ± 0.93. Aqueous extract of bark showed highest value of % DPPH° (92.92 ± 0.08 %).


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