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Industrial Crops and Products 50 (2013) 408– 413

Contents lists available at ScienceDirect

Industrial Crops and Products

journa l h om epa ge: www.elsev ier .com/ locate / indcrop

ntioxidant and antibacterial activities and composition of Brazilianpearmint (Mentha spicata L.)

odrigo Scherer ∗, Mayara Fumiere Lemos, Mariana Fumiere Lemos,ésika Coimbra Martinelli, João Damasceno Lopes Martins, Ary Gomes da Silva

niversidade Vila Velha, Rua Comissário José Dantas de Melo no. 21, Boa Vista, Vila Velha, ES CEP: 29102-770, Brazil

r t i c l e i n f o

rticle history:eceived 16 March 2013eceived in revised form 27 June 2013ccepted 3 July 2013

a b s t r a c t

Antioxidant and antibacterial activities and the composition of Brazilian spearmint (Mentha spi-cata) extracts were evaluated. They were obtained by maceration with methanol, acetone, anddichloromethane, and the essential oil was obtained by hydrodistillation. The antioxidant activity wasdetermined by antioxidant activity index (AAI), and the antimicrobial activity was evaluated by the diffu-

eywords:AIint

ntimicrobial

sion method and by determination of minimum inhibitory concentration against Staphylococcus aureusand Escherichia coli. Phenolic compounds were determined by the Folin–Cioucalteu method, and theessential oil composition was identified by GC/MS. The methanolic extract showed a higher content oftotal phenolic compounds and stronger antioxidant activity, while only the essential oil showed antibac-

ompo).

PPH terial activity. The major c(2.3%), and myrcene (2.1%

. Introduction

Many plants are used as spices to enhance food flavor andre consumed in small quantities, contributing in low levels tohe nutritional value of the diet. However, as they are secondary

etabolism compounds that may have pharmacological activity,here is currently a growing interest in plant extracts as sources ofntimicrobial and antioxidant compounds as a means of avoidingotential problems caused by excessive consumption of syntheticdditives. The current literature presents several studies on the bio-ogical activity of plant extracts as anti-tumor agents (Kaileh et al.,007), anti-inflammatories and analgesics (Bose et al., 2007), anti-ungals (Korukluoglu et al., 2008), antibacterials and antioxidantsSingh et al., 2011; Castilho et al., 2012), among others.

Mentha spicata L., commonly called spearmint, belongs to theamiaceae family, genus Mentha, which comprises about 25–30pecies originating in Europe. It is one of Brazil’s most cultivatedarieties of spearmint and is well adapted to the subtropical cli-ate. The interest in cultivating Mentha is mainly related to the

ommercial importance of its essential oil, which is among the 10ost traded in the world. The oil is used in many industries, includ-

ng pharmaceuticals, cosmetics, food, and chemicals. Spearmints also known for its ability to improve memory (Adsersen et al.,006). Besides being a stimulant (Papachristos and Stamopoulos,

∗ Corresponding author. Tel.: +55 27 3421 2072; fax: +55 27 3421 2049.E-mail addresses: rscherersm@yahoo.com.br, rodrigo.scherer@uvv.br

R. Scherer).

926-6690/$ – see front matter © 2013 Elsevier B.V. All rights reserved.ttp://dx.doi.org/10.1016/j.indcrop.2013.07.007

unds of the essential oil were carvone (67%), limonene (14.3%), muurolene

© 2013 Elsevier B.V. All rights reserved.

2002), it has several biological uses, such as in insecticides (Samarthand Kumar, 2003), antimicrobials (Ozgen et al., 2006), antioxi-dants (Choudhury et al., 2006), antispasmodics, and anti-platelets(Tognolini et al., 2006). The different species of Mentha presentconsiderable diversity in their essential oil chemical composition.For example, in M. spicata, the essential oil is rich in carvone andhas a characteristic smell of spearmint (Jirovetz et al., 2002), whilein Mentha piperita, menthol is the main component (Singh et al.,2011). In addition to such natural variation in composition amongplant species, the aromatic and pharmacological properties of theplant extracts can be significantly affected by soil and climaticconditions, as well as by the season and the time the plant mate-rial is collected (Viuda-Martos et al., 2008; Shanjani et al., 2010;Butkiene and Mockute, 2011). Considering the economic potentialof spearmint and the fact that the composition of plants may beaffected by soil and climatic conditions, and the lack of any studyon spearmint grown in the state of Espírito Santo (Brazil), the objec-tive of this study was to evaluate the composition, the antioxidantactivity, and the antimicrobial activity of extracts and essential oilsof spearmint (M. spicata) grown and purchased in the city of VilaVelha (ES/Brazil).

2. Materials and methods

2.1. Materials

Samples of spearmint (M. spicata) were purchased from pro-ducers in Vila Velha (ES, Brazil). All tests were performedusing only the aerial parts of the plant. The free radical 2,

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-diphenyl-1-picril-hydrazyl (DPPH), BHT, ferulic acid, caffeic acid,hlorogenic acid, gallic acid, Folin–Ciocalteau reagent, sodiumarbonate, and thiazolyl blue tetrazolium bromide (MTT) wereurchased from Sigma–Aldrich company (USA). The brain–heart

nfusion broth (BHI) and Mueller–Hinton agar were purchased fromiMed Laboratories Pvt. (Mumbai, India). DMSO (dimethyl sulfox-

de), methanol, acetone and dichloromethane were purchased frometec (Rio de Janeiro, Brazil). Saturated alkanes std. (C7–C30) wasurchased from Supelco (USA).

.2. Proximate analysis

Chemical analysis of the samples was performed according tohe official AOAC method (1995) (n = 7).

.3. Extracts and essential oil

Plant samples were dried in a tray drier with air circulationt 45 ◦C and ground to powder in a mill. Extracts were obtainedsing acetone, dichloromethane, and methanol as solvents. Mac-ration was done with 20 g of powdered dried plant in 100 mL ofach solvent. After 7 days under periodic agitation, the extractsere filtered through filter paper, and the residue was extracted

gain, with 100 mL of the respective solvents for 24 h. Both fractionsere then mixed and evaporated to dryness at 40 ◦C under vacuum.

he essential oil was obtained by hydrodistillation using Clevengerxtractor with aerial parts from spearmint “in nature”. The sam-les were crushed with ultra-pure water in a blender before beingransferred to the distillation flask. After extraction, the essentialil was transferred to a glass vial, and its purification was madey separation of the remnant water by freezing, and the essentialil, which was kept in liquid phase, was drained from the vial. Allxtracts were stored in amber bottles at 5 ◦C until analysis.

.4. GC/FID and GC/MS analysis

The identification of the essential oil components was carriedut by high resolution gas chromatography analysis, in the finehemistry laboratory accredited to ISO/IEC 17025, at Tommasinalítica (Vila Velha, ES, Brazil). A Thermo Scientific TraceUltra gashromatograph coupled with a Thermo Scientific DSQII quadrupoleass spectrometer (identification) and Thermo Scientific Focus

as chromatograph coupled with FID detector (quantification)ere used. The compounds were separated in a DB-5 fused silica

apillary column 30 m × 0.25 mm × 0.25 �m film thickness (J&Wcientific, Folson, USA). Helium was the Carrier gas at a flow ratef 1.0 mL/min. The analyses were performed using splitless injec-ion at 220 ◦C. The oven temperature program used was 60–240 ◦Ct 3 ◦C/min, and the final temperature was held for 7 min. TheC/MS interface and FID detector were maintained at 240 ◦C and50 ◦C, respectively. The oil was dissolved in hexane (2 mg/mL)or the analyses. The MS data were obtained in the scan mode35–400 m/z), and Kovats retention indices (KI) were determined bynjection of standard hydrocarbon solutions (C7–C30). The compo-ents were identified by comparison with data from the literatureAdams, 1995) and with the profiles from the NIST Mass Spectralibrary (version 2.0, 2005), and by injection of pure compounds,hen available.

.5. Total phenolic compounds

The total phenolics content was determined using the

olin–Ciocalteau reaction. An aliquot of 0.5 mL of a methano-ic solution of dry extracts (1.5 mg/mL) was added to 2.5 mL ofolin–Ciocalteau reagent diluted with water (1/10). After 5 min,.0 mL of 7.5% sodium carbonate was added and stirred vigorously

Products 50 (2013) 408– 413 409

in a vortex mixer. The mixture was then incubated for 2 h in the darkat room temperature. The absorbance at 740 nm was measured andconverted to the phenolics content according to a calibration curvemade with gallic acid.

2.6. Antioxidant activity

The antioxidant activity of the extracts and standards wasdetermined through radical scavenging activity by using 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) according to Scherer andGodoy (2009). Aliquots of 0.1 mL of methanol solutions of samplesor standards in different concentrations were added to 3.9 mL of amethanol solution of DPPH. DPPH solutions were prepared by dis-solving 24.5 mg in 500 mL of methanol. The blank sample consistedof 0.1 mL of methanol added to 3.9 mL of DPPH solution. The testswere carried out in triplicate. After a 90 min incubation period atroom temperature in the dark, the absorbance was measured at517 nm. The radical scavenging activity was calculated as follows:I % = [(Abs0 − Abs1)/Abs0] × 100, where Abs0 was the absorbanceof the blank and Abs1 was the absorbance in the presence of thetest compound at different concentrations. The IC50 (concentra-tion providing 50% inhibition) was graphically calculated using acalibration curve in the linear range by plotting the extract con-centration vs the corresponding scavenging effect. The antioxidantactivity was expressed as the antioxidant activity index (AAI), cal-culated as follows: AAI = final concentration of DPPH (�g/mL)/IC50(�g/mL). The assays were carried out in triplicate, and all the sam-ples, the standard, and the DPPH solutions were prepared daily.All solutions of standards and extracts were prepared in 50 mLvolumetric flasks.

2.7. Antibacterial activity

The antibacterial activity was conducted by the diffusionmethod and determination of minimum inhibitory concentra-tion (MIC) against Staphylococcus aureus (ATCC 25923) andEscherichia coli (ATCC 8739). The final bacteria concentrationwas adjusted at the First point of McFarland’s scale in order toachieve 108 CFU/mL in saline (0.85%). In the diffusion method, usingMueller–Hinton solid medium (38 g/L), the strains were inoculatedusing sterile swabs. Then cavities were made by glass tubes with adiameter of approximately 6 mm, and 50 �L samples (2 mg/mL inDMSO and saline – 0.85%) were added and incubated at 37 ◦C for24 h.

To determine the minimum inhibitory concentration (MIC), theextracts were prepared at a concentration of 4.0 mg/mL in dimethylsulfoxide (DMSO) and subsequently diluted with saline (0.85%) touse. The final concentration of cells was adjusted by the scale ofMcFarland 1 in the order of 108 CFU/mL. To each well, 100 �L ofculture medium (BHI 3.7%), 100 �L of sample, and 100 �L of inocu-lums were added. In all plates, positive and negative controls (sixwells of each) were inserted. In the positive controls, 100 �L of cul-ture medium, 100 �L of inoculum, and 100 �L of saline (0.85%) wereadded. In the negative controls, 100 �L of culture medium, 100 �Lof the respective extract, and 100 �L of saline (0.85%) were added.The extracts were evaluated from 0.67 to 0.09 mg/mL at final con-centrations. After the inoculum addition, the plates were incubatedat 36 ◦C for 24 h, and then 50 �L of the indicator MTT (0.1% in saline0.85%) was added. After 4-h incubation, the MIC was determined asthe lowest concentration that inhibits the bacterian visible growth

given by the MTT (dead cells were not stained). The absorbancewas monitored at 590 nm in microplate reader Thermo Plate (TP-Reader), and the inhibition rate was determined by the followingformula: inhibition % = ((Abs B − Abs A)/Abs B) × 100, where “Abs

410 R. Scherer et al. / Industrial Crops and Products 50 (2013) 408– 413

Table 1Proximate composition of the aerial parts of spearmint (Mentha spicata).

Protein (g/100 g) Fat (g/100 g) Carbohydratre (g/100 g)

2.3 ± 0.0 0.4 ± 0.1 9.6

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Table 2Spearmint essential oil composition.

Compound CAS no. RT KI 1 KI 2 %**

Pinene (alpha) 80-56-8 5.67 936 939 0.51Sabinene 3387-41-5 6.78 976 976 0.14Pinene (beta) 127-91-3 6.88 979 980 0.69Myrcene 123-35-3 7.17 989 991 2.08Limonene 138-86-3 8.57 1031 1031 14.34linalool 78-70-6 11.12 1100 1098 0.48Ocimene (Allo) 7216-56-0 12.35 1127 1129 0.03Ocimene (Neo-Allo) 3016-19-1 12.63 1140 1142 0.11Santolinyl acetate 79507-88-3 14.08 1174 1171 0.34Terpin-4-ol 562-74-3 14.39 1181 1177 0.07Dihydro carvone (trans) 5948-04-9 15.20 1199 1200 1.03Carveol (trans) 1197-07-5 16.00 1219 1217 0.79Carvone 99-49-0 17.69 1246 1242 67.08Carvone oxide (trans) 39067-90-8 18.53 1278 1277 0.03Dihydro carveol acetate 20777-49-5 20.53 1325 1325 0.06Elemene (delta) 20307-84-0 20.82 1332 1339 0.05Carvyl acetate* 1205-42-1 21.95 1359 1362 0.24Carvone oxide (cis)* 35178-55-3 22.01 1360 1363Copaene (alpha) 3856-25-5 22.64 1375 1376 0.09Bourbonene (beta) 5208-59-3 22.99 1383 1384 1.18Elemene (beta) 515-13-9 23.24 1388 1391 0.38Gurjunene (alpha) 489-40-7 23.93 1404 1409 0.21Caryophyllene (beta) 87-44-5 24.47 1418 1418 1.76Gurjunene (beta) 17334-55-3 24.87 1428 1432 0.19Humulene (alpha) 6753-98-6 25.89 1453 1454 0.63Muuorola-4(14),5-iene (cis) – 26.18 1460 1460 0.66Muurolene (gamma) 30021-74-0 26.99 1480 1477 2.29Bicyclogermacrene 24703-35-3 27.53 1492 1494 0.33Calamenene (cis) 72937-55-4 28.52 1518 1521 0.72Cadinene (alpha) 24406-05-1 29.15 1534 1538 0.10Muurol-5-en-beta-ol (cis) – 29.55 1545 1545 0.02Muurol-5-en-4-alpha-ol (cis) – 29.93 1554 1554 0.03Germacrene D-4-ol 74841-87-5 30.70 1574 1574 0.11Cubenol (1,10-di-epi) 73365-77-2 32.19 1612 1614 0.29Cadinol (epi-alpha) 11070-72-7 33.19 1639 1640 0.05Cadinol (alpha) 481-34-5 33.69 1652 1653 0.21

Total 97.32

RT: retention time (GC/MS); KI 1: experimental; KI 2: literature (Adams, 1995); –:

quantification of free radical scavenging activity. The reaction isbased on the decrease of the purple color that occurs when thenitrogen atom of the DPPH is reduced by receiving a hydrogen atomfrom the antioxidant component (Brand-Williams et al., 1995).

Table 3Yield and concentration of total phenols in extracts of spearmint.

Solvent Yield of extraction (g/100 g) TFC mg/g of dry extract

ME 5.96 76.32 ± 3.42a

AC 1.88 37.84 ± 1.16b

DI 2.72 Nd

Sample Moisture (g/100 g) Ash (g/100 g)

Aerial parts 86.0 ± 0.4 1.7 ± 0.0

” is the absorbance of the sample and “Abs B” is the absorbance ofhe positive control.

.8. Statistical analysis

Data were analyzed using ANOVA/Tukey (p < 0.05) by the soft-are StatisticaTM 6.0 Statsoft, Inc.

. Results and discussion

.1. Proximate analysis

Spearmint is commonly used to give a pleasant flavor and aromao many foods and beverages, but is used in small quantities, andoes not interfere with nutritional values, however, there are noeports about its composition. Table 1 shows the values of thehemical composition of a 100 g sample, which presents contentigh in moisture and low in fat, as characteristic of foliaceous plants.

.2. Essential oil composition

In the essential oil chromatographic analysis (Fig. 1), 37 com-ounds were identified, representing 97.32% of the total oilTable 2). The spearmint essential oil was characterized by theominant presence of carvone (67.1%), in agreement with otheruthors, such as Kokkini et al. (1995), who identified carvoneccurring at 68.4% in samples collected in Greece, Chauhan et al.2009), who found it at 76.65% in samples collected in India, and

arer et al. (2011), who found it at 48.4% in samples collected inurkey. Also, in essential oil from M. spicata collected in Montene-ro (Europe), Sokovic and Van Griensven (2006) reported carvones the major constituent (49.52%), followed by mentone (21.92%)nd limonene (5.77%). However, in the present study, limonene wasound at 14.3%, but mentone was not found. This variation in theomposition of essential oil may be attributed to various factorsn growing conditions, such as temperature, humidity, radiation,limate, and soil type. Other significant constituents in the essen-ial oil of M. spicata included muurolene (2.29%), myrcene (2.08%),eta-caryophyllene (1.76%), beta-bourbonene (1.18%), and trans-ihidrocarvone (1.03%).

.3. Total phenolic compounds

Phenolic compounds are the major class of natural antiox-dants present in plants and are usually quantified using theolin–Ciocalteu method. The results obtained in the determinationf total phenolics, expressed as gallic acid equivalents (GAE) perram of dry extract, are presented in Table 3. Methanol had theighest yield of crude extract and phenolic contents (p < 0.05), whenompared with those from acetone and dichloromethane. Dormant al. (2003) reported that total phenolic content in different vari-ties of Mentha is about 128–230 mg/g of extract in gallic acidquivalent, and the main phenolic compounds found in extractsf M. spicata were eriocitrin, luteolin, rosmarinic acid, and caffeic

cid. However, the amount of phenolic compounds found in thisork is below that range. The total phenolic content in the extract

btained with dichloromethane could not be determined becauset was not solubilized in methanol.

not found.* Co-elution.

** Quantified by GC/FID.

3.4. AAI results

Various methods are used to determine the antioxidant activ-ity of plant extracts and pure substances, though the DPPH(2,2-diphenyl-1-picril-hydrazyl) method is the most used in the

HD 0.04 15.16 ± 0.50c

AC: acetone; DI: dichloromethane; ME: methanol, HD: essential oil; Nd: not evalu-ated. TFC: total phenolic compounds. Different letters in the same column representsignificant difference (p < 0.05).

R. Scherer et al. / Industrial Crops and Products 50 (2013) 408– 413 411

RT: 0,00 - 67,02

0 5 10 15 20 25 30 35 40 45 50 55 60 65e (min)

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10

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30

40

50

60

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80

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22,9928,527,17 32,1915,20 33,6922,014,00 11,12 48,97 63,3956,0746,4743,31

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Fig. 1. GC/MS of spearmint essen

ecause of the difficulty of comparing the antioxidant activity ofifferent plants, Scherer and Godoy (2009) not only proposed aew index to express the strength of natural antioxidant com-ounds (AAI), but also established a ranking for antioxidant activityf plant extracts in relation to AAI values: weak antioxidant activ-ty (AAI < 0.5), moderate (0.5–1.0), strong (1.0–2.0), and very strongAAI > 2.0). Recently, Deng et al. (2011) proposed a new index, thentioxidant activity unit (AAU) for that purpose based on the workf Scherer and Godoy (2009), and the authors reported that theyound some limitations in the AAI, such as lack of a precise defini-ion and high values absorbance (between 2 and 3), which woulde outside the range of accuracy according to Beer’s law.

Regarding the first observation of Deng et al. (2011), the def-nition is clear and simple, just a relationship between the massatio of radical used in the assay, with the mass of extract neededo reduce 50% of the radicals, generating a constant called the AAI.n the second point, according to the equation of the Beer’s law,

he absorbance is proportional to the optical path: the larger theize of the cell, the greater the absorbance value for the same con-entration. Scherer and Godoy (2009) evaluated the linearity ofifferent concentrations of DPPH in methanol using 1 cm2 cell, andhe results showed that the absorbance values were at the range of.05–2.0, being the recommended concentration of 0.1 mM, whichenerates an absorbance near 1.0. In this study, the linearity of theolutions of DPPH (0.05–1.5 mM) was repeated using two differentpectrophotometers with 1 cm2 cells (Hach DR 5000: certificate ofSO/IEC 17025 no. EVO-1882/11 and T80+ UV/VIS, PG Instrumentstd), and the results agreed with Scherer and Godoy (2009).

In order to get satisfactory reproducibility on the AAI results,ome precautions were taken, such as the use of calibrated ana-ytical balance and the use of high quality volumetric flasks torepare the standard solutions. Other precautions included pro-ecting standard solutions and plant extracts from the excessivexposure to light, verifying the purity degree and validity of mate-ials, and finally, training the laboratory workers.

The results of mint extracts are presented in Table 4. Galliccid showed the highest value of the AAI (p < 0.05), followed byaffeic acid. The other compounds, such as chlorogenic acid, fer-lic acid, and BHT, had lower values of AAI. However, all of themere considered strong antioxidants, according to the classifica-

ion. The differences in activity can be explained by the relationshipetween structure and activity, because the structure of phenolicompounds is a key determinant for the oxidation of free radicalsRice-Evans et al., 1996). The greater the number of the aromatic

l diluted with hexane (2 mg/mL).

ring hydroxylations handled, the greater the activity of phenolicacids (Scherer and Godoy, 2009), as in the case of gallic acid, whichshowed the highest values of AAI. The replacement of the hydroxylgroup in the aromatic ring by a methoxyl group reduces the valueof AAI, which explains caffeic acid being more active than ferulicacid.

Spearmint methanolic extract showed a strong antioxidantactivity, according to the proposed classification (AAI = 2.08). Onthe other hand, the essential oil and acetone and dichloromethaneextracts showed no ability to reduce free radical DPPH. Thus, it issuggested that the polar components of the plant have a higherantioxidant capacity than the non-polar compounds by the DPPHmethod, as shown by the solvent methanol having a higher polar-ity index (ability to make hydrogen bonds). Kanatt et al. (2007)confirmed the antioxidant activity of M. spicata during radiationprocessing of lamb, in that the aqueous extract was able to retardlipid peroxidation and give a pleasing aroma and flavor to the meat.According to Tepe et al. (2007), plants of the Lamiaceae family arevery rich in phenolic compounds, and these have been shown tohave antioxidant activity, which agrees with this work, since themethanol extract showed a higher content of phenolic compoundsand higher antioxidant activity.

3.5. Antibacterial activity

In tests carried out by the diffusion method, formation of inhi-bition halos indicating the absence of antimicrobial activity of allextracts for both microorganisms was not observed. On the otherhand, in the MIC determination, the essential oil showed strongactivity against strains of E. coli and S. aureus. However, the extractsobtained with methanol, acetone, and dichloromethane did notshow significant activity. The microorganism S. aureus was moresensitive to essential oils, and a final concentration of 0.67 mg/mLshowed 100% inhibition of growth. For E. coli, the highest concen-tration tested (0.67 mg/mL) showed 51.3% inhibition, and thereforethe MIC could not be determined. Boukhebti et al. (2011) reportedthat Gram-positive bacteria are more sensitive to essential oils thanGram-negative ones, due to the more complex cell wall in Gram-negative, agreeing with this work. Higher concentrations were notevaluated because the green coloration of the extracts affect theabsorbance reading and may produce false results, so it is neces-

sary to make the clarification by the removal of chlorophyll withoutaffecting the results. The fact that the essential oil of spearmint hasantimicrobial activity only in the MIC determination tests may be

412 R. Scherer et al. / Industrial Crops and Products 50 (2013) 408– 413

Table 4Antioxidant activity index (AAI).

DPPH* Day I* Day II Day III Média SD

r2 IC50 AAI r2 IC50 AAI r2 IC50 AAI AAI AAI

Galic acid 0.980 1.6 24.3 0.995 1.65 23.61 0.996 1.51 25.81 24.59a 1.12Caffeic acid 0.984 2.1 18.6 0.978 1.81 21.4 0.98 1.97 19.76 19.91b 1.43Chlorogenic acid 0.982 4.29 9.1 0.985 3.65 10.68 0.96 4.12 9.47 9.75c 0.83Ferulic acid 0.963 6.13 6.36 0.989 5.91 6.6 0.996 6.18 6.31 6.42d 0.15BHT 0.976 9.38 4.24 0.994 9.11 4.37 0.999 12.44 3.2 3.94 e 0.64ME 0.992 17.99 2.25 0.991 21.7 1.99 0.994 21.86 1.98 2.08 e 0.15AC – – – – – – – – – – –DI – – – – – – – – – – –HD – – – – – – – – – – –

r2: coefficient of linearity, IC50: expressed in �g/mL, SD: standard deviation, –: values were not found; ME: methanol, AC: acetone, DI: dichloromethane; HD: essential oil.Different letters in the same column represent significant difference (p < 0.05).

dbctcptpoa2

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4

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* Tests carried out on 3 different days.

ue to the difficulty of diffusion of compounds through the medium,ecause the compounds are mostly of low polarity, whereas theulture medium has polar character. Regarding the composition ofhe spearmint essential oil, carvone was found at 67% of the totalomposition. According to Kadoglidou et al. (2011), this monoter-enoid has antimicrobial activity, and this may be responsible forhe antibacterial activity of spearmint’s essential oil reported in theresent study. There is evidence in the literature that the essentialils of some plants of the Lamiaceae family have a moderate to goodntibacterial activity (Sokovic and Van Griensven, 2006; S arer et al.,011).

Some works with plants of the genus Mentha have reportedhat the crude extracts present higher antioxidant activity than thessential oils; on the other hand, the essential oils were more effec-ive against foodborne spoilage or pathogenic bacteria than thextracts obtained by extraction with solvents, as observed in theresent study (Hussain et al., 2010; Teixeira et al., 2012; Dhifi et al.,012). In another study, the essential oil showed strong antimi-robial activity against all 30 microorganisms tested, whereas theethanol extract remained almost inactive. In contrast, the extract

howed much better activity than the essential oil in antioxidantctivity assays employed (Gulluce et al., 2007). This fact can bexplained by the presence of some non-volatile phenolic com-ounds in the crude extracts obtained by solvents, which areotential antioxidants, and generally not present in the essentialil, such as hydroxycinnamic acids, flavonoids, and others. On thether hand, the essential oils are composed mainly of terpenoids,.e. carvone, which exhibit strong antimicrobial activity, and theironcentrations are low in the crude extracts.

. Conclusion

The methanolic extract showed high amount of phenolic com-ounds and strong antioxidant activity. These results suggest it cane used as an alternative or substitute for the synthetic antiox-

dants that present harmful effects to humans. Furthermore, thepearmint essential oil, which has great commercial value in Brazil,howed good antimicrobial activity due to the high concentrationf carvone (67%), suggesting that it can be used in the cosmetics andood industries. The essential oil can contribute to the preservationf products, and may develop pleasant sensory characteristics.

cknowledgements

We acknowledge the Laboratory Tommasi Analítica (Vila Velha,S, Brazil) for the cooperation in the chromatographic analysis; andUNADESP for the Research Grant of Dr. Rodrigo Scherer and Dr. Aryomes da Silva.

References

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