phytochemicalanalysisandantimicrobial/antioxidant

Research ArticlePhytochemical Analysis and AntimicrobialAntioxidantActivity of Cynodon dactylon (L) Pers RhizomeMethanolic Extract

Samira Savadi1 Mohsen Vazifedoost 1 Zohre Didar1 Mohammad Mahdi Nematshahi2

and Eisa Jahed3

1Department of Food Science and Technology Neyshabur Branch Islamic Azad University Neyshabur Iran2Department of Food Science and Technology amp Young Researchers and Elite Club Sabzevar Branch Islamic Azad UniversitySabzevar Iran3Department of Food Science and Technology Urmia University Urmia Iran

Correspondence should be addressed to Mohsen Vazifedoost mvazifedoostiau-neyshaburacir

Received 3 December 2019 Revised 27 January 2020 Accepted 5 March 2020 Published 1 April 2020

Academic Editor Jorge Barros-Velazquez

Copyright copy 2020 Samira Savadi et alis is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

A proper use of medicinal plants requires accurate scientific information and understanding of their chemical constituents etherapeutic effects in the plants are due to the chemical compounds therein Cynodon dactylon (L) Pers of the family Poaceae is aperennial pan-tropical species of grass which is a well-known traditional medicine and has a renowned position for treatment ofmany symptoms and diseases e chemical composition free radical scavenging activity and antimicrobial properties of themethanolic extract of C dactylon rhizomes against selected bacterial and fungal strains were investigated using disk-diffusionmethod e results indicated that the major fatty acid structures of C dactylon methanolic extract were palmitic acid (3640)oleic acid (2826) and linoleic acid (1701) Alpha-tocopherol (15139mgmiddotkgminus1) and sitosterol (319962mgmiddotkgminus1) were the maintocopherols and sterols respectively According to the instrumental analysis the total phenolic compounds of methanolic extractwere equal to 91708mgmiddotkgminus1 and the main compound was hydroquinone (6689) Antioxidant activity of the methanolic extractat concentrations of 100ndash1000 ppm was 981ndash6787 which is significantly different from the 200 ppm synthetic antioxidant(BHT) with free radical scavenging activity equal to 4893 (plt 005) e antimicrobial study revealed that the methanolicextract of C dactylon rhizomes was effective against all of the bacterial pathogens and the antibacterial activity increased with anincrease in the concentration of extract erefore the largest zone of inhibition was observed against Bacillus cereus(183plusmn 04mm) and Escherichia coli (168plusmn 05mm) at 1000 ppm e highest resistance was observed with Pseudomonasaeruginosa (inhibition zone of 128plusmn 015mm) Methanolic extract at 200 ppm had no effect against fungus Aspergillus nigerHowever at 1000 ppm concentration an inhibition zone with a diameter of 144plusmn 045mm was formed

1 Introduction

Medicinal plants play a very important role in the devel-opment of alternative drugs without the adverse effects of thesynthetic drugs [1] Plants and natural products form thebasis of both modern and traditional medicines and now-adays they are widely used in the production of commer-cially produced drugs Scientific and reliable reportsindicated that about 25 of prescribed medicines worldwideare taken from herbs [2] Plant essential oils extracts and

various kinds of secondary metabolites are known as sub-stances with antimicrobial and antioxidant properties withlittle or no toxic effects which are vital in the management ofmany diseases [2 3] Actually secondary metabolites withtremendous biological potential known as phytochemicalsare ubiquitously present in plants and now are used as themainstay of drug development [4]

As reported in the literature according to an estimationof the WHO about 80 percent of the worldrsquos population inLatin America Africa Asia and the Middle East in

HindawiJournal of Food QualityVolume 2020 Article ID 5946541 10 pageshttpsdoiorg10115520205946541

particular relies on herbs for its primary healthcare needs[5] ese herbs have minimal side effects and recentlymillions of dollars have been invested in pharmaceuticalplants by pharmaceutical companies in order to producenatural medicines extracted from herbs [5 6] e mainreasons for using medicinal herbs are as follows (i) theycorrespond more closely to the patientrsquos ideology (ii) theyreduce concerns about the side effects of synthetic medi-cines (iii) they are more affordable (iv) they satisfy a desirefor more personalized health care and (v) they also allowmore peoplersquos access to health information [2]

A substantial body of the literature has reported themultitude pharmacological applications of plant extractsand the compounds isolated from plant extracts [7] Har-nessing the biological potential of medicinal plants repre-sents a sterling opportunity for the development of noveltherapeutic candidates [8 9] e bioactive plant extracts area promising source of many drugs For example berberine(Berberis) and Quinine (Cinchona) are the antibiotics ob-tained from plants which are highly effective against bac-teria (Escherichia coli and Staphylococcus aureus) [10]anks to the climate diversity a vast diversity of wildbioactive plants is found in Iran erefore it is possible toobtain herbal extracts in large quantities on industrial scale

Cynodon dactylon (family Poaceae) commonly known asdhub doob or harialil other common names include durba(Bengali) garikoihallu (Kanarese) garikagaddi (Telugu)durua or haritali (Sanskrit) dhubkhabbal (Punjabi) durua(Marathi) and arugampul (Tamil) C dactylon (L) Pers is aweed plant found in many regions such as East Africa AsiaAustralia and Southern Europe [10] It is a perennial grasswhich is used as forage as medicinal plant and for desertgreening e roots grow quickly leading to the rapid spreadof C dactylon (L) Pers It is mostly found in cereal fieldsfarms road shoulders and green spaces and parks e plantis propagated through seeds and rhizomes When crop seedsgerminate and plants are established in the field rhizomes ofC dactylon (L) Pers can grow in hard soils between roots ofother crop plants to produce new plants [11] Photochemicalanalyses have shown that C dactylon (L) Pers containsflavonoids alkaloids glycosides terpenoids triterpenoid es-ters saponins tannins resins phytosterols reducing sugarscarbohydrates proteins volatile oils and fixed oils [6 12]According to Unani system of medicine C dactylon has asharp and hot taste with a good odor e aerial parts andrhizomes of the plant have cardioprotective action and an-tibacterial antimicrobial antioxidant wound-healing anti-diabetic and diuretic effects [10 13]

C dactylon is used by traditional healers for purifying theblood diarrhea gonorrhea conjunctivitis anuria biliousnessitches and stomach ache [11] e literature survey alsoreveals that the dried extracts of C dactylon aerial parts wereinvestigated for CNS activities in rat [5] Other essentialfunctions of this plant include analgesic and antipyreticantiulcer antihypertensive antihysteric antipyretic antibi-otic antikidney stone antiviral antipsychotic antigonorrhealinfection as well as hypoglycemic agent [11 14 15] Studies onlab animals have shown that methanolic extract of C dactylondecreases the level of lipid peroxides It was also revealed that

the methanolic extract ofC dactylon had an antioxidant effecton COLO 320 DM cells a colon cancer cell line and the levelsof antioxidant enzymes [6]

Few studies have been conducted on medicinal and an-timicrobial properties of C dactylon (L) Pers [10 13ndash15] andfurther studies must be carried out to confirm its antioxidantand antimicrobial properties Chemical constituents of themethanolic extract of the rhizomes of C dactylon (L) Persnative to Iran were identified in this study for the first time andits antioxidant and antimicrobial properties were investigated

2 Materials and Methods

21 Materials A sufficient amount of C dactylon rhizomeswas collected from Mashhad Iran e extraneous partswere removed and the rhizomes were washed immediatelydried and ground Lyophilized ampoules containing stan-dard strains of P aeruginosa E coli B cereusA niger and Sepidermidis were obtained from the Regional Center forBacteria and Fungi Collection of the Iranian Research Or-ganization for Science and Technology Other high-puritychemicals used in this study were bought from Sigma In-corporation and Merck Company (Darmstadt Germany)

22 Methods

221 Preparation of the Extract of C dactylon Rhizome

(1) Maceration Extraction For maceration extraction therhizomes of C dactylon were ground using a KenwoodCG100 grinder sieved and then mixed with methanol at aratio of 1 10 (wtvol) e resulting mixture was placed on ahotplate stirrer at 250 rpm and ambient temperature for24 h e mixture was then filtered under vacuum using aBuchner funnel and Whatman filter paper No 1 To removethe methanol the mixture was concentrated by a rotaryevaporator (LABOROTA 4000) at 35degC and the extract waseventually dried under vacuum in a drier at 40degC and kept ina closed impermeable container at 4degC until further use [16]

(2) Microwave-Assisted Extraction After mixing the plantpowder withmethanol (1 10 wtv) the sample was soaked inmethanol for 60min For better extraction the resultingmixture was irradiated using a microwave oven for 2 4 and6min at a constant 150W power e mixture was thenfiltered under vacuum using a Buchner funnel and What-man filter paper No 1 To remove the methanol the mixturewas concentrated by a rotary evaporator (LABOROTA 4000)at 35degC and the extract was eventually dried under vacuumin a drier at 40degC and kept in a closed impermeable containerat 4degC until further use [17]

(3) Ultrasound-Assisted Extraction e plant powder wasmixed with the methanol solvent (1 10 wtv) and extractionwas performed in an ultrasonic bath (Elma) at 80 kHz for 1015 and 20min e mixture was then filtered under vacuumusing a Buchner funnel and Whatman filter paper No 1 Toremove the methanol the mixture was concentrated by arotary evaporator (LABOROTA 4000) at 35degC and the

2 Journal of Food Quality

extract was eventually dried under vacuum in a drier at 40degCand kept in a closed impermeable container at 4degC untilfurther use [18]

23 Experiments

231 Identification of the Chemicals in the Extract e fattyacid profile of the sample was determined by gas-liquidchromatography (GLC) and reported based on relativepercentages for levels [19] Tocopherol compounds weredetermined by high-performance liquid chromatography(HPLC Yang Lin Co) and sterol levels in the samples weremeasured by gas chromatography (GC) [19] In additionphenolic components were identified by using GC-MS andthrough comparing retention times of compounds with theWiley 299 Nist 107 and Nist 21 Libraries

232 Oxidative Stability Index Oxidative stability indexwas calculated from the following equation based on theconcentration of C18 unsaturated fatty acids

Cox value 1 C181( 1113857 + 103 C182( 1113857 + 216 C183( 11138571113858 1113859

100

(1)

Here C181 C182 and C183 represent oleic linoleic andlinolenic acids respectively [19]

233 Total Phenolic Content Total phenolic content of theextract was measured using FolinndashCiocalteu reagent Forthis purpose 25mL of 02N FolinndashCiocalteu reagent wasadded to 05mL of each extract (10mgmL) and 2mL of75 gL sodium carbonate solution was then added after5min Absorbance by the mixture was read against the blanksolution at 760 nm after 2 h Gallic acid was used as astandard to plot the calibration curve e total phenoliccontent was reported in terms of mg gallic acidg extracteexperiments were performed in triplicate and the averagewas reported [20]

234 Free Radical (DPPH) Scavenging Activity e stableradical compound 22-diphenyl-1-picryl-hydrazyl-hydrate(DPPH) was used to evaluate the free radical scavengingactivity of the methanolic extract of Cynodon dactylonrhizomes e ability to donate hydrogen atoms or electronsby the different compounds in the extract was evaluated bythe discoloration rate or reduction in absorbance by thepurple DPPH solution in methanol Two milliliters of eachextract concentration was added to 2mL of 0004 DPPHsolution in methanol After 90min of storage in the dark atroom temperature absorbance by the sample at 517 nm wasread against the controle free radical (DPPH) scavengingactivity was calculated using the following equation [21]

I Ablank minus Asample

Ablanktimes 100 (2)

Here Ablank and Asample represent absorbance by the extract-free control and the extract respectively For the

comparison 100 ppm concentration of the synthetic anti-oxidant butylhydroxyanisole (BHA) was used forcomparison

235 Antimicrobial Properties of the Extract A syringe filterwith the pore diameter of 045 microm was used to sterilize theextract e extract was obtained from the rhizomes ofCynodon dactylon at different concentrations (200 600 and1000 ppm) and the controls were separately prepared inmethanol A fresh culture of each microorganism was re-quired to prepare the microbial suspensions erefore 24 hprior to the test the inclined nutrient agar was inoculatedwith the microorganisms from the stored cultures eculture was then washed with Ringerrsquos solution to preparethe microbial suspension A small volume of the microbialsolution was poured into a tube containing sterile Ringerrsquossolution and its turbidity was measured by a spectropho-tometer at 530 nm e solution was diluted by Ringerrsquossolution until the turbidity reaches 05 of McFarland stan-dard e resulting suspension needed to contain15times108 CFUmL of bacteria Antimicrobial activity of themethanolic extract of C dactylon was evaluated using agardisk-diffusion method To this end one loop of the standardculture of each strain was cultured on a sterile Muel-lerndashHinton agar medium Paper discs (diameter of 6mm)sterilized in an autoclave at 121degC for 15min were im-pregnated by different concentrations of the methanolicextract of C dactylon ey were then placed on the surfaceof culture medium using sterile forceps and fixed with a littlepressure on the culture medium It is noteworthy that an-timicrobial activity of the standard gentamicin disks (10 microgdisc) was separately evaluated in Petri dishes After incu-bation for 24 h at 37degC the diameter of the nongrowth halowas measured using a caliper (INSTAR China) with aprecision of 001mm All experiments were performed intriplicate [22]

24 Statistical Analysis A completely randomized statisticaldesign was used to analyze the results e data were sta-tistically analyzed on SAS Comparison of the means witheach other and the control was carried out using Duncanrsquostest at a probability level of 5 (plt 005) Diagrams wereplotted in Microsoft Excel 2010

3 Results and Discussion

31 6e Effects of Extraction Method on Extraction Yieldree extraction methods using methanol as solvent wereused in this study to extract the active ingredients in therhizomes of C dactylon including ultrasound-assisted ex-traction microwave-assisted extraction at constant powerand different times and maceration for 24 h According tothe results the different extraction methods had differentyields so that the maceration and the 10min ultrasoundextraction showed the highest and lowest extraction yields of109 and 289 respectively (Figure 1) e results indicatedthat extraction yield improved with the increase of time inboth microwave- and ultrasound-assisted extraction

Journal of Food Quality 3

methods By increasing the time from 10 to 20min in theultrasound-assisted extraction the yield improved from 289to 329 Furthermore the yield of the microwave-assistedextraction improved from 381 in 2min to 433 in 6minese observed differences were statistically significant(plt 005)

Plants contain many compounds with different struc-tures Extraction of these components depends on multiplefactors and the most important of which are the solvent andthe extraction method Selection of the solvent and theextraction method depend on the different parts of the plantand their constituent materials It is very difficult to select acertain solvent for each class of plant compounds becausethere are other substances along with these compounds thataffect the solubility of the compounds of interest [23] Itshould be noted that a high extraction yield for an extractdoes not necessarily mean a high content of active ingre-dients According to the literature despite the higher yield ofan extraction method than another one the contents of theextracted active ingredients such as phenolic compoundsmay be lower in the extract with the higher yield [4]

Traditionally maceration is used for extraction of plantmaterials ere are numerous studies on the use of mac-eration to extract various plant compounds including an-tioxidants For example Hypericum perforatum L extractobtained by maceration shows a good antioxidant activity[24] Antioxidant activity of corn silk [25] Diospyros lotusPyrus boissieriana and Leontodon hispidus extracted bymaceration has also been reported [26] ere are reports ofnumerous pharmacological effects of the methanolic extractof Juglans regia L obtained by maceration extraction [27]ere are also reports on the antioxidant activity of the aerialparts of Allium paradoxum and Ferula gummosa Boissextracted by maceration [27 28] All these studies indicatethe good performance of this classic extraction methodMany studies have also been conducted to compare differentextraction methods and their yields for extracting activeingredients ese studies indicate that there are insignifi-cant differences between the extraction methods For ex-ample Soxhlet ultrasound and maceration methods wereused for extracting Cucumis melo leaves e results showed

that the phenolic content of the extract obtained by Soxhletextraction was higher e extract obtained by the Soxhletmethod also showed the highest iron chelating activity Allthree extraction methods were effective in extracting anti-oxidant compounds without any significant differencesbetween them [28] Similar results were obtained regardingthe antioxidant activity of Vicia faba so that all three ex-traction methods effectively extracted antioxidant com-pounds without any significant differences between them[29]

32 Chemicals Found in theMethanolic Extract of C dactylonRhizomes

321 Structure of Fatty Acids Table 1 lists the chemicalstructures of the fatty acids in the methanolic extract of Cdactylon rhizomes extracted by maceration Clearly themajor fatty acids in the methanolic extract of C dactylonrhizomes were saturated fatty acids including palmitic acid(C160 4036) and stearic acid (C180 410) and the un-saturated fatty acids including oleic acid (2826) and linoleicacid (1701) e levels of saturated and unsaturated fattyacids in the methanolic extract of C dactylon rhizomes werealmost the same Saturated fatty acids (SFAs) monoun-saturated fatty acids (MUFAs) and polyunsaturated fattyacids (PUFAs) comprised 4739 2898 and 2187 of thetotal fatty acids respectively (Table 1)

e PUFASFA ratio of 1073 was obtained for themethanolic extract of C dactylon rhizomes is ratio isconsidered as a measure of unsaturated oils and fats andtheir tendency for autoxidation of lipids [30] ereforecomparison of the oxidative stability index of 305 for theextract with the PUFASFA ratio indicates a good agreementbetween these two oxidation indices According to theseoxidation indices the methanolic extract of C dactylon has agood resistance to oxidation

322 Tocopherol Compounds Tocopherols are a very im-portant nonsoap part of plant extracts and oils Because ofthe antioxidant activity of tocopherols these naturally oc-curring compounds in plant essential oils are extremelyvaluable for human health ey protect fats and oils againstdegradation through reaction with free radicals anddirecting oxidation reactions to their end stages is makestocopherols very valuable for human health [31]

According to the results the total tocopherol content inthe methanolic extract of C dactylon rhizomes was27947mgkg with α-tocopherol accounting for the largestpart (15139mgkg) Levels of both c- and β-tocopherol were12808mgkg however Δ-tocopherol was not detected(Table 2) erefore one can conclude that the methanolicextract of C dactylon rhizomes is a valuable tocopherolsource that can be used in food and pharmaceuticalindustries

323 Evaluation of Sterol Compounds Plant sterols alsoknown as phytosterols comprise more than 50 of nonsoap

c c

Microwave (150W)

Maceration

Ultrasound (80kHz)

b

a

e d d

0

2

4

6

8

10

12

14

2 4 6 8 10 15 20

Yiel

d (

)

Extraction method

Figure 1 Effects of different methods on extraction yield of Cdactylon rhizomes (different letters indicate statistically significantdifferences between groups as determined by the Duncan test(plt 005))


compounds Plant sterols are triterpenoids and more thanone hundred different types of them have been identified innature ese metabolites are found in all plant tissueshowever they exist with a higher frequency in seeds [32]Phytosterols have antioxidant antibacterial anti-inflam-matory and wound-healing properties [33] According tothe instrumental results five sterol compounds with a totalcontent of 986624mgkg were identified in the methanolicextract of C dactylon rhizomes Beta-sitosterol stigmasteroland campesterol comprise 319962mgkg (3243 of totalsterol content) 232547mgkg (2357) and 202356mgkg(2051) of the total sterols respectively Among theidentified sterols the lowest sterol content of 46075mgkg(467) was that of brassicasterol (Table 3)

Sterols are lipophilic components of the membraneplaying a key role in its fluidity ey are necessary forvarious cell functions ey also have extensive biologicalactivities including anti-inflammatory anticancer antioxi-dant and antibacterial activities Moreover these metabo-lites are able to reduce blood serum cholesterol level [34]erefore C dactylon seeds can be considered as richsources of medicinal sterols and a good alternative tocholesterols in food diet According to the Mangathayaruet al (2009) reports C dactylon steroid saponins have somebiological and pharmacological activities including anti-bacterial diuretic and hypocholesteremic characteristics Itwas also found that the fresh juice of C dactylon has im-munomodulatory and DNA-protective activity [33]

324 Identification and Measurement of PhenolicCompounds Nowadays phenolic compounds in plants areconsidered as one of the best natural antioxidant sourcesPhenolic compounds form a special group of secondary

metabolites that play a key role in the protection of tissuesagainst oxidation effects of free radicals oxygen and otheractive species ereby they prevent the occurrence ofnumerous diseases such as inflammatory diseases cancerdiabetes myocardial infarction and Alzheimerrsquos and Par-kinsonrsquos diseases [2] Given the direct relationship betweenantioxidant effects and levels of phenolic compounds itseems necessary to identify and measure phenolic com-pounds in the methanolic extract of C dactylon rhizomesPhenolic compounds in the methanolic extract were iden-tified by GCMS and their relationship with changes in theextract concentration in terms of mg gallic acidg extract isshown in Table 4 and Figure 2 respectively According to theinstrumental results 17 chemicals with hydroxyl groupswere identified Among them hydroquinone was the mostabundant phenolic compound (6689) with two OHgroups in C1 and C4 positions of the aromatic ringe othernotable phenolic compounds were thymol (123) levo-glucosenone (248) phytol (112) vanillic acid (135)and syringic acid (098) (Table 4)

In a similar study Jananie et al [35] analyzed thechemicals in the aqueous extract of C dactylon leavesAmong the identified compounds glycerine (3849) andphytol (489) were the major and minor phenolic com-pounds respectively [35] In another study Kumar andChandel [11] analyzed C dactylon leaf extract and identified24 chemicals e main compounds were glycerine (385)thymol (115) ethyl glucopyranoside (842) and phytol(45) [11] Various factors affect the levels and types ofphenolic compounds in plant tissues including geneticfactors solar radiation soil conditions stage of maturity atharvest time weather and environmental conditionspostharvesting operation and storage conditions [36]According to the literature on extraction of phenoliccompounds from plant tissues an increase in the extractiontemperature significantly increases extraction yield of

Table 2 Tocopherol compounds in the methanolic extract of Cdactylon rhizomes

Tocopherol compounds Level (mgkg)α-Tocopherol 15139c- and β-tocopherol 12808Δ-Tocopherol NDTotal 27947

Table 3 Sterol compounds in the methanolic extract of C dactylonrhizomes

Sterol compounds (mgkg) PercentageCampesterol 202356 2051Stigmasterol 232547 2357Brassicasterol 46075 467Beta-sitosterol 319962 3243Δ5 Avenasterol mdash NDΔ7 Stigmasterol 185781 1883Δ7 Avenasterol mdash NDTotal sterol compounds 986624 mdashND nondetectable

Table 1 Fatty acid profiles of the methanolic extract of C dactylonrhizomes

Fatty acid Level ()(C120)Lauric acid 020(C140)Myristic acid 093(C150)Pentadecanoic acid 017(C160)Palmitic acid 4036(C161)Palmitoleic acid 037(C170)Margaric acid 021(C180)Stearic acid 410(C181t)Oleic acid 013(C181c)Oleic acid 2826(C182t)Linoleic acid 028(C182c)Linoleic acid 1701(C183)Linolenic acid 458(C200)Arachidonic acid 064(C201)Paulinic acid 019(C220)Behenic acid 040(C240)Lignoceric acid 038Other fatty acids 113Saturated fatty acids (SFAs) 4739Monounsaturated fatty acids (MUFAs) 2898Polyunsaturated fatty acids (PUFAs) 2187PUFASFA 1073Oxidative stability index 305


phenolic compounds as higher temperature softens planttissues and weakens interactions between phenolic com-pounds proteins and polysaccharides (and consequentlyremoves more phenolic compounds from the plant to thesolvent) [37]

e total phenolic content in the methanolic extract ofCdactylon rhizomes was 91708mgg extract As seen inFigure 2 when the extract concentration rises from 100 to1000 ppm the level of phenolic compounds in the extractincreases from 064 to 2742mg gallic acidg extract eobserved increases were significant at the probability level of95 for all extract concentrations compared to control with018mg gallic acidg extract (plt 005)is shows the strongdependence of phenolic compounds in the methanolic ex-tract of C dactylon rhizomes on the concentration so thatthe level of phenolic compounds significantly improved withincreases in extract concentration (Figure 3) and led toimproved antioxidant properties

According to the literature the high level of phenoliccompounds is the main reason for the enhanced antioxidantactivity of some of the extracts including polar extracts

Evidences show that there is a positive correlation betweenthe level of phenolic compounds and antioxidant activity ofplants Moreover it seems that phenolic compounds that arefound abundantly in plants with high antioxidant activitycan be extracted mainly through plant extracts [38] e keyrole of phenolic compounds as free radical scavengers hasbeen reported in several papers [39 40] It is noteworthy thatphenolic compounds effectively act as hydrogen donors andthereby as an effective antioxidant Unver et al [41] ex-amined the antioxidant activity and total phenol content ofmethanolic extracts of some plants and found a direct re-lationship between the total phenol content and antioxidantactivity of plant extracts [41] is is consistent with ourresults

325 Free Radical (DPPH) Scavenging Activity e activityof the stable DPPH radical is used to evaluate free radicalscavenging activity in different sampleseDPPH is a violetstable hydrophilic free radical with the highest absorbance at515ndash517 nm When receiving electrons from reducing

Table 4 Phenolic compounds identified in the methanolic extract of C dactylon rhizomes

Phenolic compound Molecular formula Retention time (min) Level ()Propanoic acid 2-oxo- C3H4O3 379 142ymol C10H14O 554 123Pantolactone C6H10O3 984 0832-Furancarboxaldehyde 5-methyl- C6H6O2 775 155Pentanoic acid 4-oxo- C5H8O3 997 075Levoglucosenone C6H6O3 1207 2483-Hydroxy-1-methylpyridinium hydroxide C6H9NO2 1377 162Phytol C20H40O 1528 112Propanedioic acid phenyl- C9H8O4 1548 128Hydroquinone C6H6O2 1604 668913-Benzenediol 5-chloro- C6H5ClO2 1741 122Benzaldehyde 3-(chloromethyl)-4-methoxy- C10H9ClO4 1863 085Ethanone 1-(4-hydroxy-3-methoxyphenyl)- C9H10O3 2015 05216-Anhydro-a-d-glucopyranose (levoglucosan) C6H10O5 2029 105Vanillic acid C8H8O4 2130 135Syringic acid C9H10O5 2494 098Dimethoxybicyclo[331]nona-24-dione C11H16O4 2697 092

e e d c

b

a

0

5

10

15

20

25

30

35

0 100 200 400 800 1000

Tota

l phe

nolic

com

poun

ds(m

gg)

Rhizome extract (ppm)

Figure 2 Total phenolic compounds at different concentrations ofthe methanolic extract of C dactylon rhizomes (different lettersindicate statistically significant differences between groups as de-termined by the Duncan test (plt 005))

fe

d

b

a

c

01020304050607080

100 200 400 800 1000 200 ppmBHT

DPP

H ra

dica

l sca

veng

ing

activ

ity (

)


Figure 3 Scavenging activity of free radicals by different con-centrations of the methanolic extract of C dactylon rhizomes incomparison with the synthetic antioxidant BHT (different lettersindicate statistically significant differences between groups as de-termined by the Duncan test (plt 005))


compounds such as phenols DPPH is converted into thecolorless compound hydrazine is structural change isassociated with reduced absorbance Compounds with suchcapability are considered antioxidants [42] Figure 3 com-pares the scavenging activity of different concentrations ofthe methanolic extract of C dactylon rhizomes with that of200 ppm of the synthetic antioxidant BHT measured byDPPH Clearly the free radical scavenging activity signifi-cantly improves with increasing concentrations of themethanolic extract of C dactylon rhizomes e scavengingactivity at 100 ppm was 981 and improved to 6787 as theextract concentration increased to 1000 ppm e scav-enging activity of the methanolic extract of C dactylonrhizomes at 200 400 and 800 ppm was 202 3236 and5442 respectively

By adding 200 ppm of the synthetic antioxidant andcomparing its scavenging activity with different concen-trations of themethanolic extract ofC dactylon it was foundthat 200 ppm of the synthetic antioxidant BHT had a lowerscavenging activity (4893) than that of 800 and 1000 ppmmethanolic extract of C dactylon rhizomes (plt 005) isindicates the high scavenging activity of the methanolicextract of C dactylon rhizomes especially at higher con-centrations e level of this parameter was consistent withthat of phenolic compounds in the methanolic extract of Cdactylon rhizomes ese results indicate that the phenoliccompounds may be responsible for antioxidant activity ofthe plant extract e antioxidant potential of the phenoliccompounds in plant extracts depends on their type andconcentration as well as the number and position of hy-droxyl groups in the aromatic ring An increase in theconcentration of phenolic compounds directly increases theability of different extracts in scavenging free radicals Due toan increase in the number of hydroxyl groups in the reactionmedium at higher concentrations of phenolic compoundsthe likelihood of hydrogen donation to free radicals andthereby scavenging activity of the extract increases [43]Furthermore these phenolic compounds are converted tophenoxyl free radicals after donating hydrogen e stabilityof these radicals may affect antioxidant capacity of phenoliccompounds as less-stability phenoxyl radicals compete withDPPH radicals for absorbing hydrogen atoms which leadsto a decrease in the entrapment of DPPH radicals [40]

e antioxidant activity of hydroalcoholic extract of theaerial parts ofC dactylonwas evaluated by different methods(DPPH radicals scavenging activity superoxide anion rad-ical scavenging nitric oxide scavenging iron chelating ca-pacity hydroxyl radical scavenging and hydrogen peroxidescavenging activity) In all these methods the extract showeda concentration-dependent scavenging activity for freeradicals Determination of superoxide anion radicals showeda maximum antioxidant activity of 9333 [44] In anotherstudy C dactylon extract was continuously extracted withhexane ethyl acetate and methanol e extract was thenconcentrated and its antioxidant activity was tested on fourcancer cell lines and a normal cell line by DPPH nitric oxideand 3-(45-dimethylthiazol-2-yl)-25-diphenyl tetrazoliumbromide (MTT) e anticancer potential of the cytotoxicextract was determined by using Annexin-fluorescein

isothiocyanate-conjugated assay in human colon adeno-carcinoma cell lines (COLO 320 DM) e results showedsignificant antioxidant and antiproliferative activities ofCynodon dactylon extracts [12]

326 Antibacterial Effects of the Methanolic Extract of Cdactylon Rhizomes e spread of drug-resistant and anti-biotic-resistant microbial strains is among major problemsin the food and drug industries Medicinal plants have re-cently received a great deal of attention from researchers forthe treatment of acute diseases due to toxic and carcinogenicproperties of chemical and synthetic compounds Accord-ingly natural antimicrobial and antioxidant compoundssuch as organic acids essential oils and plant extracts can beconsidered as good and safe alternatives in foods [44] In thisresearch the antimicrobial effects of the methanolic extractof C dactylon native to Iran were evaluated for the first timeTo this end multiple strains of Gram-positive bacteria (Bcereus) Gram-negative bacteria (P aeruginosa E coli) and afungus (A niger) were selected to determine the effects ofdifferent extract concentrations (200 400 and 1000 ppm) onthe nongrowth of these microorganisms using the disk-diffusion method and nongrowth halo diameter (in mm)Afterwards these effects were compared with that of gen-tamicin as a standard antibiotic (Table 5)

All the concentrations of methanolic extract of C dac-tylon were effective in preventing the growth of both Gram-positive and Gram-negative bacteria and formation of thenongrowth halo e antioxidant activity of the extractincreased with an increase in the extract concentrationwhich was due to an increase in the phytochemical com-pounds in the extract e Gram-positive bacterium B ce-reus was more sensitive to the extract (a maximumnongrowth halo diameter of 183plusmn 04mm) whereas theGram-negative bacterium P aeruginosa showed the highestresistance to the extract with a nongrowth halo diameter of226plusmn 028mm e extract showed the least effect on Paeruginosa so that no halo was formed around the disk emethanolic extract of the C dactylon rhizomes with aconcentration of 200 ppm had no impact on A niger still ahalo with a diameter of 144plusmn 045mm was formed as theextract concentration increased to 1000 ppm However thiswas significantly lower than the effect of gentamicin (Ta-ble 5) Among the studied microorganisms extract-free puremethanol (negative control) did not affect P aeruginosa andA niger however a non-growth halo with a diameter of341plusmn 018 18plusmn 008 and 23plusmn 01mm was formed for Ecoli B cereus and S epidermidis respectively which wassignificantly lower than those of the treatments containingthe methanolic extract of C dactylon rhizomes andgentamicin

Hydrophobicity is an important property of plant ex-tracts which enables them to rupture cell membrane andremove vital molecules and ions of bacteria out of the cellsand ultimately kill bacteria by forming bonds to the lipidlayer of the cell membrane of bacteria and mitochondria[14] In a similar study the antibacterial activity ofhydroalcoholic extract of Cynodon dactylon against two


Gram-positive bacteria (S aureus and S albus) and twoGram-negative bacteria (E coli and P aeruginosa) wasevaluated using the disc-diffusion (zone of inhabitation) andthe dilution (minimum inhibitory concentration) methodse results showed that the hydroalcoholic extract of Cdactylon had an effective antibacterial activity e resultsregarding the minimum inhibitory concentrations indicatedthe sensitivity of all bacterial strains to C dactylon extract[45] In another study the antimicrobial activity of crude Cdactylon extracts obtained from seven different solvents(acetone chloroform diethyl ether ethanol ethyl acetateand n-pentane) against some pathogens was investigatedusing the disk-diffusion method e results showed theextensive antimicrobial activity of ethanol (7ndash10mm) andethyl acetate (7ndash12mm) extracts against all bacterial path-ogens Both methanol and acetone extracts showed a sig-nificant antimicrobial activity against B cereus and Bsubtilis whereas the chloroform extract showed an anti-microbial activity against S pyogenes Ethyl acetate andethanol extracts with inhibition zones of 8ndash15mm and8ndash13mm respectively showed a reasonable antimicrobialactivity However no significant antimicrobial activity wasfound against A niger [14]

Suresh et al [46] examined theantibacterial activity ofC dactylon leaf extract against pathogenic bacteria (Bsubtilis S aureus E coli K pneumonia and S aerugi-nosa) using the agar disk-diffusion method [46]According to the results the chloroform extract of Cdactylon leaves showed a significant antibacterial activityagainst all tested bacteria e chloroform extract with aconcentration of 75 μLmL formed a larger inhabitationzone than those with concentrations of 25 and 50 μLmLMost bacteria however were resistant to the aqueousextract of C dactylon In a different in vivo study theantiviral activity of C dactylon extract produced on a largescale on white spot syndrome virus (WSSV) was inves-tigated in Penaeus monodon To this end the C dactylonextract was mixed with artificial pellet feed with a con-centration of 1 or 2 e results showed the significanteffect of C dactylon extract on prevention of WSSV in-fection without mortality [47] In another researchRahman (2014) by studying the antimicrobial activity ofthree different extraction of C dactylon against somepathogen bacteria reported that the hot and cold aqueousextracts of this herb had antimicrobial activity against allthe test Gram-positive and Gram-negative bacteria in-dicating broad-spectrum activity of the extract for or-ganisms While the effect of methanol extract was not

significant this is probably due to the differences in themethod of extraction and nonuse of ultrasound in thepreparation of methanol extract [5]

According to the results of this research and of thosereported in other studies one can conclude that themethanolic extract of C dactylon rhizomes is able to inhibitthe growth of pathogenic and spoilage bacteria in foodsus it is recommended to use it as a natural preservativeand flavoring agent in food products

4 Conclusion

Given the growing awareness in consumers of the effects ofsynthetic antioxidants on public health and the ever-in-creasing prevalence of drug resistance in bacteria moreattention has been paid to finding ways of preventing drugresistance and also developing effective drugs with less sideeffects Medicinal plants such as C dactylon which grow indifferent regions of Iran have received a great deal of at-tention in this regard e results showed that the meth-anolic extract of C dactylon rhizomes especially at higherconcentrations was able to scavenge free radicals isfeature was due to higher levels of antioxidant tocopheroland phenolic compounds e extracts with concentrationsof 800 and 1000 ppm showed higher scavenging activity thanthe extract with concentration of 200 ppm of the syntheticantioxidant BHT Furthermore the methanolic extract of Cdactylon rhizomes showed a wide range of activities againstboth Gram-positive and Gram-negative bacteria and againstthe mold A niger as well e antibacterial activity of themethanolic extract of C dactylon arising from phyto-chemical compounds confirms its application as a thera-peutic drug in folk medicine erefore biologicalcompounds that are identified in C dactylon can be used toproduce antimicrobial drugs for the treatment of variousdiseases such as eye infections pneumonia and gonorrheae results of this study showed that the methanolic extractof C dactylon rhizomes could be considered an antibacterialagent and a potential source of antibiotics for the treatmentof bacterial infections Due to the high antioxidant activity ofthis plant extract it can be used as a good alternative tosynthetic antioxidants for preserving food materials andedible oils

Data Availability

e data used to support the findings of this study are in-cluded within the article

Table 5 Inhibitory activity of the methanolic extract of C dactylon rhizome on the growth of microorganisms

Inhabitation zone diameter (mm)A niger S epidermidis B cereus E coli P aeruginosa Concentration (ppm)000plusmn 00d 23plusmn 01e 18plusmn 008e 334plusmn 018d 000plusmn 00d Control (0)000plusmn 00d 91plusmn 02d 112plusmn 015d 1025plusmn 04c 73plusmn 02c 200126plusmn 03c 125plusmn 035c 148plusmn 025c 153plusmn 03ab 104plusmn 03b 600144plusmn 045b 157plusmn 04ab 183plusmn 04b 168plusmn 05a 128plusmn 015a 1000226plusmn 028a 1752plusmn 017a 208plusmn 05a 161plusmn 01a 000plusmn 00d GentamicinlowastDifferent letters indicate statistically significant differences between groups as determined by the Duncan test (plt 005)


Conflicts of Interest

e authors declare that there are no conflicts of interestwith respect to the authorship andor publication of thisarticle

Acknowledgments

e authors thank the management of Islamic Azad Uni-versity-Neyshabur Branch for their financial support and thedirector of central lab of the university for the help with theexperiments

References

[1] S Andrade M J Ramalho J A Loureiro and M D PereiraldquoldquoNatural compounds for Alzheimerrsquos disease therapy asystematic review of preclinical and clinical studiesrdquo Inter-national Journal of Molecular Sciences vol 20 p 41 2019

[2] I F F Benzie and S Wachtel-Galor Herbal Medicine Bio-molecular and Clinical Aspects CRC PressTaylor amp FrancisGroup Boca Raton FL USA 2nd edition 2011

[3] K R R Rengasamy H Khan S Gowrishankar et al ldquoe roleof flavonoids in autoimmune diseases therapeutic updatesrdquoPharmacology amp 6erapeutics vol 194 pp 107ndash131 2019

[4] R Dutt V Garg N Khatri and A K Madan ldquoPhyto-chemicals in anticancer drug developmentrdquo Anti-CancerAgents in Medicinal Chemistry vol 19 no 2 pp 172ndash1832019

[5] S Rahman Cynodon Dactylon Antimicrobial Potential ofCrude Extract as Valuable Medicinal Plant BRAC UniversityBangladesh India 2014

[6] P D A E Al-Snafi ldquoA review on chemical constituents andpharmacological activities of Coriandrum sativumrdquo IOSRJournal of Pharmacy (IOSRPHR) vol 6 no 7 pp 17ndash42 2016

[7] A B Oyenihi and C Smith ldquoAre polyphenol antioxidants atthe root of medicinal plant anti-cancer successrdquo Journal ofEthnopharmacology vol 229 pp 54ndash72 2019

[8] M Ayaz F Ullah A Sadiq et al ldquoSynergistic interactions ofphytochemicals with antimicrobial agents potential strategyto counteract drug resistancerdquo Chemico-Biological Interac-tions vol 308 pp 294ndash303 2019

[9] P Maher ldquoe potential of flavonoids for the treatment ofneurodegenerative diseasesrdquo International Journal of Molec-ular Sciences vol 20 no 12 p 3056 2019

[10] D Venkatachalam B S avamani and K MuddukrishniahldquoAntimicrobial activity and phytochemical analysis of aerialparts of Cynodon dactylonrdquo International Journal of AcademicResearch and Development vol 3 no 3 pp 116ndash121 2018

[11] E Chandel and B Kumar ldquoAntimicrobial activity and phy-tochemical analysis of Cynodon dactylon a reviewrdquo WorldJournal of Pharmacy and Pharmaceutical Sciences vol 4no 11 pp 515ndash530 2015

[12] A Albert-Baskar and S Ignacimuthu ldquoChemopreventiveeffect of Cynodon dactylon (L) Pers extract against DMH-induced colon carcinogenesis in experimental animalsrdquo Ex-perimental and Toxicologic Pathology vol 62 no 4pp 423ndash431 2010

[13] P K Rai D Jaiswal D K Rai B Sharma and G WatalldquoAntioxidant potential of oral feeding ofcynodon dactylo-nextract on diabetes-induced oxidative stressrdquo Journal ofFood Biochemistry vol 34 no 1 pp 78ndash92 2010

[14] S Abdullah J Gobilik and K P Chong ldquoIn vitro antimi-crobial activity of Cynodon dactylon (L) pers (bermudas)against selected pathogensrdquo Developments in SustainableChemical and Bioprocess Technology vol 16 pp 227ndash2372013

[15] A S Kumar K Gnananath D Kiran A M Reddy andC Raju ldquoAntidiabetic activity of ethanolic extract of Cynodondactylon root stalks in streptozotocin induced diabetic ratsrdquoInternational Journal of Advances in Pharmaceutical Researchvol 2 no 8 pp 418ndash422 2011

[16] M Mazandarani S Makri and G R Bajian ldquoEvaluation ofphytochemical and antioxidant activity in different parts ofHeracleum gorganicum rech F in Golestan Province North ofIranrdquordquo Iranian Journal of Plant Physiology vol 2 no 2pp 381ndash388 2011

[17] M Mirzapour M Hamedi and M Rahimipanah ldquoSunfloweroil stabilization by Persian walnut leaves extract during ovenstorage testrdquo Food Science and Technology Research vol 16no 5 pp 443ndash446 2010

[18] R D Sotillo M Hadley and E T Holm ldquoPhenolics inaqueous potato peel extract extraction identification anddegradationrdquordquo Journal of Food Science vol 59 no 2pp 649ndash651 1994

[19] R Farhoosh J Tavakoli and M H H KhodaparastldquoChemical composition and oxidative stability of kernel oilsfrom two current subspecies of Pistacia atlantica in IranrdquoJournal of the American Oil Chemistsrsquo Society vol 85 no 8pp 723ndash729 2008

[20] A Ordonez J Gomez M Vattuone andM lsla ldquoAntioxidantactivities of Sechium edule (jacq) swartz extractsrdquo FoodChemistry vol 97 no 3 pp 452ndash458 2006

[21] A Dashipour R Khaksar H Hosseini S Shojaee-Aliabadiand K Ghanati ldquoPhysical antioxidant and antimicrobialcharacteristics of carboxymethyl cellulose edible film coop-erated with clove essential oilrdquo Zahedan Journal of Research inMedical Sciences vol 16 no 8 pp 34ndash42 2014

[22] S Mohammadi Amlashi and B Babakhani ldquoExamining theantibacterial activity of Artemisia dracunculus L extractsusing different methods of extractionrdquo International Journalof Molecular and Clinical Microbiology vol 6 no 1pp 629ndash634 2016

[23] S F Nabavi M A Ebrahimzadeh S M NabaviM Mahmoudi and S K Rad ldquoBiological activities of Juglansregia flowersrdquo Revista Brasileira de Farmacognosia vol 21no 3 pp 465ndash470 2011

[24] H Fathi ldquoAntioxidant and free radical scavenging activities ofHypericum perforatum Lrdquo International Journal of Forest Soiland Erosion vol 3 no 2 pp 68ndash72 2013

[25] M A Ebrahimzadeh F Pourmorad and S Hafezi ldquoAnti-oxidant activities of Iranian corn silkrdquo Turkish Journal ofBiology vol 32 no 1 pp 43ndash49 2008

[26] M A Ebrahimzadeh S Eslami S M Nabavi S F Nabaviand B Eslami ldquoAntioxidant and antihemolytic activitiesofLeontodon hispidosrdquo Biotechnology amp BiotechnologicalEquipment vol 24 no 4 pp 2127ndash2131 2010

[27] S M Nabavi M A Ebrahimzadeh S F Nabavi B Eslamiand A A Dehpour ldquoAntioxidant and antihaemolytic activ-ities of Ferula foetida regel (Umbelliferae)rdquo European Reviewfor Medical and Pharmacological Sciences vol 15 no 15pp 157ndash164 2011

[28] M A Ebrahimzadeh M Askari and M Forouzani ldquoEval-uation of three methods for the extraction of antioxidantsfrom Cucumis melo L fruit and leavesrdquo International Journalof Forest Soil and Erosion vol 3 no 3 pp 95ndash99 2013


[29] Z Hashemi andM A Ebrahimzadeh ldquoEvaluation of methodsfor the extraction of antioxidants from Vicia faba L bean andhullsrdquo Latin American Applied Research vol 44 pp 203ndash2082014

[30] F Sahin M Gulluce D Daferera et al ldquoBiological activities ofthe essential oils and methanol extract of Origanum vulgaressp vulgare in the Eastern Anatolia region of Turkeyrdquo FoodControl vol 15 pp 549ndash557 2004

[31] R Swetie Ch Raesh and S Arun ldquoAntioxidant potential ofmint (Mentha Spicata L) in radiationprocessed lamb meatrdquoFood Chemistry vol 100 no 2 pp 451ndash458 2007

[32] M J Lagarda G Garcıa-Llatas and R Farre ldquoAnalysis ofphytosterols in foodsrdquo Journal of Pharmaceutical and Bio-medical Analysis vol 41 no 5 pp 1486ndash1496 2006

[33] K Mangathayaru M Umadevi and C U Reddy ldquoEvaluationof the immunomodulatory and DNA protective activities ofthe shoots of Cynodon dactylonrdquo Journal of Ethno-pharmacology vol 123 no 1 pp 181ndash184 2009

[34] Y Boutte and M Grebe ldquoCellular processes relying on sterolfunction in plantsrdquo Current Opinion in Plant Biology vol 12no 6 pp 705ndash713 2009

[35] R K Jananie V Priya and K Vijayalakshmi ldquoDeterminationof bioactive components of Cynodon dactylon by GC-MSanalysisrdquo New York Science Journal vol 4 2011

[36] S Khanizadeh L Ding R Tsao et al ldquoPhytochemical dis-tribution among selected advanced apple genotypes devel-oped for fresh market and processingrdquo Journal of FoodAgriculture and Environment vol 1 no 2 2007

[37] J Shi J Yu J Pohorly J C Young M Bryan and Y WuldquoOptimization of the extraction of polyphenols from grapeseed meal by aqueous ethanol solutionrdquo Food Agriculture ampEnvironment vol 1 no 2 pp 42ndash47 2003

[38] C Chatchawan B Soottawat H Jakul and S NattigaldquoAntioxidant components and properties of five long- grainedrice bran extracts from commercial available cultivars inailandsrdquo Food Chemistry vol 111 no 3 pp 636ndash641 2008

[39] V Katalinic M MIlos T Kulisic and M Jukic ldquoScreening of70 medicinal plant extracts for antioxidant capacity and totalphenolsrdquo Food Chemistry vol 94 no 4 pp 550ndash557 2006

[40] M eriault S Caillet S Kermasha and M Lacroix ldquoAn-tioxidant antiradical and antimutagenic activities of phenoliccompounds present in maple productsrdquo Food Chemistryvol 98 no 3 pp 490ndash501 2006

[41] A Unver D Arslan M M Ozcan andM Akbulut ldquoPhenoliccontent and antioxidant activity of some spicesrdquo WorldApplied Sciences Journal vol 6 pp 373ndash377 2009

[42] S Kim S-C Ko Y-S Kim et al ldquoDetermination of Curcumalonga L (Turmeric) leaf extraction conditions using responsesurface methodology to optimize extraction yield and anti-oxidant contentrdquo Journal of Food Quality vol 2019 pp 1ndash82019

[43] J Wang X Cao H Jiang Y Qi K Chin and Y YueldquoAntioxidant activity of leaf extracts from different Hibiscussabdariffa accessions and simultaneous determination fivemajor antioxidant compounds by LC-Q-TOF-MSrdquo Mole-cules vol 19 no 12 pp 21226ndash21238 2014

[44] M J Saharkhiz M Sattari GhR Goudarzi and R Omidbygyldquoe effect of antibacterial herb Tanacetum parthenium LrdquoJournal-Iranian Medicinal and Aromatic Plants Researchvol 24 pp 47ndash55 2008

[45] S Renu and N B Prakash ldquoScreening of antibacterial activityof hydroalcoholic extract of Cynodon dactylon (L)rdquo Inter-national Journal of Research in Ayurveda and Pharmacyvol 3 no 6 pp 827ndash829 2012

[46] K Suresh P Deepa R Harisaranraj and V Vaira AchudhanldquoAntimicrobial and phytochemical investigation of the leavesof Carica papaya L Cynodon dactylon L Pers Euphorbia hirtaLMelia azedarach L and Psidium guajava Lrdquo EthnobotanicalLeaflets vol 12 pp 1184ndash1191 2008

[47] A S Hameed G Balasubramanian M SarathiC Venkatesan and J omas ldquoOral administration of an-tiviral plant extract of Cynodon dactylon on large scale pro-duction against white spot syndrome virus (WSSV) inPenaeus monodonrdquo Journal of Aquaculture vol 279 pp 2ndash52008


particular relies on herbs for its primary healthcare needs[5] ese herbs have minimal side effects and recentlymillions of dollars have been invested in pharmaceuticalplants by pharmaceutical companies in order to producenatural medicines extracted from herbs [5 6] e mainreasons for using medicinal herbs are as follows (i) theycorrespond more closely to the patientrsquos ideology (ii) theyreduce concerns about the side effects of synthetic medi-cines (iii) they are more affordable (iv) they satisfy a desirefor more personalized health care and (v) they also allowmore peoplersquos access to health information [2]

A substantial body of the literature has reported themultitude pharmacological applications of plant extractsand the compounds isolated from plant extracts [7] Har-nessing the biological potential of medicinal plants repre-sents a sterling opportunity for the development of noveltherapeutic candidates [8 9] e bioactive plant extracts area promising source of many drugs For example berberine(Berberis) and Quinine (Cinchona) are the antibiotics ob-tained from plants which are highly effective against bac-teria (Escherichia coli and Staphylococcus aureus) [10]anks to the climate diversity a vast diversity of wildbioactive plants is found in Iran erefore it is possible toobtain herbal extracts in large quantities on industrial scale

Cynodon dactylon (family Poaceae) commonly known asdhub doob or harialil other common names include durba(Bengali) garikoihallu (Kanarese) garikagaddi (Telugu)durua or haritali (Sanskrit) dhubkhabbal (Punjabi) durua(Marathi) and arugampul (Tamil) C dactylon (L) Pers is aweed plant found in many regions such as East Africa AsiaAustralia and Southern Europe [10] It is a perennial grasswhich is used as forage as medicinal plant and for desertgreening e roots grow quickly leading to the rapid spreadof C dactylon (L) Pers It is mostly found in cereal fieldsfarms road shoulders and green spaces and parks e plantis propagated through seeds and rhizomes When crop seedsgerminate and plants are established in the field rhizomes ofC dactylon (L) Pers can grow in hard soils between roots ofother crop plants to produce new plants [11] Photochemicalanalyses have shown that C dactylon (L) Pers containsflavonoids alkaloids glycosides terpenoids triterpenoid es-ters saponins tannins resins phytosterols reducing sugarscarbohydrates proteins volatile oils and fixed oils [6 12]According to Unani system of medicine C dactylon has asharp and hot taste with a good odor e aerial parts andrhizomes of the plant have cardioprotective action and an-tibacterial antimicrobial antioxidant wound-healing anti-diabetic and diuretic effects [10 13]

C dactylon is used by traditional healers for purifying theblood diarrhea gonorrhea conjunctivitis anuria biliousnessitches and stomach ache [11] e literature survey alsoreveals that the dried extracts of C dactylon aerial parts wereinvestigated for CNS activities in rat [5] Other essentialfunctions of this plant include analgesic and antipyreticantiulcer antihypertensive antihysteric antipyretic antibi-otic antikidney stone antiviral antipsychotic antigonorrhealinfection as well as hypoglycemic agent [11 14 15] Studies onlab animals have shown that methanolic extract of C dactylondecreases the level of lipid peroxides It was also revealed that

the methanolic extract ofC dactylon had an antioxidant effecton COLO 320 DM cells a colon cancer cell line and the levelsof antioxidant enzymes [6]

Few studies have been conducted on medicinal and an-timicrobial properties of C dactylon (L) Pers [10 13ndash15] andfurther studies must be carried out to confirm its antioxidantand antimicrobial properties Chemical constituents of themethanolic extract of the rhizomes of C dactylon (L) Persnative to Iran were identified in this study for the first time andits antioxidant and antimicrobial properties were investigated

2 Materials and Methods

21 Materials A sufficient amount of C dactylon rhizomeswas collected from Mashhad Iran e extraneous partswere removed and the rhizomes were washed immediatelydried and ground Lyophilized ampoules containing stan-dard strains of P aeruginosa E coli B cereusA niger and Sepidermidis were obtained from the Regional Center forBacteria and Fungi Collection of the Iranian Research Or-ganization for Science and Technology Other high-puritychemicals used in this study were bought from Sigma In-corporation and Merck Company (Darmstadt Germany)

22 Methods

221 Preparation of the Extract of C dactylon Rhizome

(1) Maceration Extraction For maceration extraction therhizomes of C dactylon were ground using a KenwoodCG100 grinder sieved and then mixed with methanol at aratio of 1 10 (wtvol) e resulting mixture was placed on ahotplate stirrer at 250 rpm and ambient temperature for24 h e mixture was then filtered under vacuum using aBuchner funnel and Whatman filter paper No 1 To removethe methanol the mixture was concentrated by a rotaryevaporator (LABOROTA 4000) at 35degC and the extract waseventually dried under vacuum in a drier at 40degC and kept ina closed impermeable container at 4degC until further use [16]

(2) Microwave-Assisted Extraction After mixing the plantpowder withmethanol (1 10 wtv) the sample was soaked inmethanol for 60min For better extraction the resultingmixture was irradiated using a microwave oven for 2 4 and6min at a constant 150W power e mixture was thenfiltered under vacuum using a Buchner funnel and What-man filter paper No 1 To remove the methanol the mixturewas concentrated by a rotary evaporator (LABOROTA 4000)at 35degC and the extract was eventually dried under vacuumin a drier at 40degC and kept in a closed impermeable containerat 4degC until further use [17]

(3) Ultrasound-Assisted Extraction e plant powder wasmixed with the methanol solvent (1 10 wtv) and extractionwas performed in an ultrasonic bath (Elma) at 80 kHz for 1015 and 20min e mixture was then filtered under vacuumusing a Buchner funnel and Whatman filter paper No 1 Toremove the methanol the mixture was concentrated by arotary evaporator (LABOROTA 4000) at 35degC and the



23 Experiments



Cox value 1 C181( 1113857 + 103 C182( 1113857 + 216 C183( 11138571113858 1113859

100

(1)





Ablanktimes 100 (2)


















c c

Microwave (150W)

Maceration

Ultrasound (80kHz)

b

a

e d d

0

2

4

6

8

10

12

14

2 4 6 8 10 15 20

Yiel

d (

)

Extraction method






















e e d c

b

a

0

5

10

15

20

25

30

35

0 100 200 400 800 1000

Tota

l phe

nolic

com

poun

ds(m

gg)



fe

d

b

a

c

01020304050607080

100 200 400 800 1000 200 ppmBHT

DPP

H ra

dica

l sca

veng

ing

activ

ity (

)
















4 Conclusion


Data Availability







Acknowledgments


References



















































23 Experiments



Cox value 1 C181( 1113857 + 103 C182( 1113857 + 216 C183( 11138571113858 1113859

100

(1)





Ablanktimes 100 (2)


















c c

Microwave (150W)

Maceration

Ultrasound (80kHz)

b

a

e d d

0

2

4

6

8

10

12

14

2 4 6 8 10 15 20

Yiel

d (

)

Extraction method






















e e d c

b

a

0

5

10

15

20

25

30

35

0 100 200 400 800 1000

Tota

l phe

nolic

com

poun

ds(m

gg)



fe

d

b

a

c

01020304050607080

100 200 400 800 1000 200 ppmBHT

DPP

H ra

dica

l sca

veng

ing

activ

ity (

)
















4 Conclusion


Data Availability







Acknowledgments


References




























































c c

Microwave (150W)

Maceration

Ultrasound (80kHz)

b

a

e d d

0

2

4

6

8

10

12

14

2 4 6 8 10 15 20

Yiel

d (

)

Extraction method






















e e d c

b

a

0

5

10

15

20

25

30

35

0 100 200 400 800 1000

Tota

l phe

nolic

com

poun

ds(m

gg)



fe

d

b

a

c

01020304050607080

100 200 400 800 1000 200 ppmBHT

DPP

H ra

dica

l sca

veng

ing

activ

ity (

)
















4 Conclusion


Data Availability







Acknowledgments


References





































































e e d c

b

a

0

5

10

15

20

25

30

35

0 100 200 400 800 1000

Tota

l phe

nolic

com

poun

ds(m

gg)



fe

d

b

a

c

01020304050607080

100 200 400 800 1000 200 ppmBHT

DPP

H ra

dica

l sca

veng

ing

activ

ity (

)
















4 Conclusion


Data Availability







Acknowledgments


References






























































4 Conclusion


Data Availability







Acknowledgments


References




















































Acknowledgments


References


















































phytochemicalanalysisandantimicrobial/antioxidant

Documents