Characterization of Satureja khuzestanica Leaf as a Herbal Medicine

Microsc. Microanal. 20, 1425–1435, 2014doi:10.1017/S1431927614013026

© MICROSCOPY SOCIETYOF AMERICA 2014

Characterization of Satureja khuzestanica Leaf as aHerbal MedicineMaryam Malmir,1 Rita Serrano,1 Ahmad Reza Gohari,2 and Olga Silva1,*

1Research Institute for Medicines and Pharmaceutical Sciences (iMed.UL), Pharmacological Sciences Group, Faculty ofPharmacy, University of Lisbon, Av. Prof. Gama Pinto, 1649-019 Lisbon, Portugal2Medicinal Plants Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, 16 Azar Avenue, 14155-6451Tehran, Iran

Abstract: Dried leaves of Satureja khuzestanica Jamzad are a popular herbal medicine and dental anestheticamong the nomadic inhabitants of southwestern Iran. The present study establishes criteria for identificationof S. khuzestanica dried whole, fragmented and powdered leaves for specification as a herbal substance usingmacroscopic and microscopic characterization. Quantitative microscopy techniques were also considered.Macroscopically leaves exhibit a grayish-green color, are broadly ovate in shape with an acute apex, attenuatebase, and ciliate margin and have a surface covered by an indumentum of glandular and non-glandular trichomes.Microscopically leaves have an isobilateral amphistomatic structure containing peltate glandular trichomesconsisting of a multiseriate stalk (five cells) and an enlarged secretory head composed of 12 cells, capitateglandular trichomes of variable morphology together with two types of non-glandular trichomes. Oval-shapedhygromorphic diacytic stomata with an adaxial stomatal index of 13.54, collateral vascular bundles consistingof xylem, and three layers of sclerenchymatous tissue close to phloem together with cluster, prismatic andraphide calcium oxalate crystals were also identified as useful pharmacognostic parameters for identification ofS. khuzestanica dried leaves.

Key words: Satureja khuzestanica leaf, herbal substance, quality control, macroscopic analysis, light microscopy,scanning electron microscopy

INTRODUCTION

The genus Satureja L. belongs to the Lamiaceae family, sub-family Nepetoideae, tribe Saturejeae, and comprises morethan 200 species of often aromatic herbs and shrubs widelydistributed in the Mediterranean area, Asia and boreal Amer-ica (Rustaiyan et al., 2004). Botanical and systematic descrip-tions of the genus Satureja have been discussed by severaltaxonomists in various flora publications (Bentham, 1876;Briquet, 1895–1897; Shishkin, 1954; Heywood & Richardson,1972; Davis, 1982; Rechinger, 1982; Doroszenko, 1985; Lee &Hedge, 1994).

Sixteen species of this genus (Satureja) have been reportedfrom Iran and nine of them, namely S. atropatana Bunge,S. sahendica Bornmüller, S. bachtiarica Bunge, S. isophyllaRechinger, S. edmondi Briquet, S. kallarica Jamzad, S. khuzes-tanica Jamzad, S. rechingeri Jamzad, and S. kermanshahensisJamzad are endemic in this country. They usually appear insmall populations in mountainous habitats (Rechinger, 1982;Jamzad, 1992, 1994, 1996, 2009, 2010).

Satureja species have been used in traditional medicineas antimicrobial, spasmolytic, cicatrisant, and diuretic agents(Gohari et al., 2005). They are well known medicinal plantswith great economic and medical importance in Iran due

to their high content of essential oil and their use in thepharmaceutical, food, perfumery, and cosmetics industries(Akbarinia & Sefidkon, 2009).

Satureja khuzestanica Jamzad, known by the commonPersian name “Marzeh Khuzestani” is one of the importantmedicinal plants among the nomadic inhabitants of south-western Iran. Traditional herbal preparations consisting ofaqueous infusion and decoction of the aerial parts (mostlyleaf) have been used as a dental analgesic and oral antiseptic(Farsam et al., 2004).

Recently a broad range of biological activities ofS. khuzestanica extracts and essential oil were investigated, andfound to have antibacterial (Seghatoleslami et al., 2009; Shahabet al., 2011; Hadian et al., 2012; Motaharinia et al., 2012), anti-fungal (Sadeghi-Nejad et al., 2010; Zarrin et al., 2010), anti-parasitic (Kheirandish et al., 2011; Sadeghi-Nejad et al., 2011;Zibaei et al., 2012), antioxidant (Abdollahi et al., 2003; Rezvanfaret al., 2008, 2010; Ahmadvand et al., 2012; Hashemi et al., 2012;Saei-Dehkordi et al., 2012), anti-diabetic (Abdollahi et al., 2003;Saadat et al., 2004; Nazari et al., 2005; Basiri et al., 2007; Vosough-Ghanbari et al., 2008; Shahsavari et al., 2009; Tavafi et al., 2011;Kaeidi et al., 2013), anti-inflammatory (Amanlou et al., 2005;Ghazanfari et al., 2006; Rezvanfar et al., 2010; Rastegarpanahet al., 2011; Shahab et al., 2011), anti-coagulant, and anti-hyperlipidemic (Nazari et al., 2005) properties. Carvacrol wasidentified as the main constituent of the essential oil (Sefidkon &Ahmadi, 2000; Farsam et al., 2004; Hadian et al., 2011).*Corresponding author. [email protected]

Received May 5, 2014; accepted July 18, 2014

mailto:[email protected]

Due to its remarkable pharmacological activities, theessential oil has been used in the pharmaceutical andfood industries in the form of a dental anesthetic and oralantiseptic drops. Capsules containing the fine powder ofdried leaves and tablets containing the extract of aerial partsare used as food supplements for their antioxidant andtriglyceride-lowering activities (Hadian et al., 2011).

In “Flora of Iran”, S. khuzestanica was described as a sub-shrub to 30 cm high, with opposite-decussate leaves, broadlyovate or orbicular (3− 5× 6− 8mm), pubescent, attenuate atthe base, and ciliate at the margin (Ghahraman, 1989).

It was noticed that S. khuzestanica is close to S. edmondiexcept for differences in stem (erect and branched inS. khuzestanica and subflexuose in S. edmondi), inflorescence(verticillasters shortly pedunculate and remote in S. khuzestanicaand subsessile and approximate in S. edmondi), and leafinternodes (2–3 mm long in S. khuzestanica and 8–10 mm inS. edmondi) (Jamzad, 1994).

Results of geographical and geological studies ofS. khuzestanica habitats indicate that the plant grows on dry,limestone rocky slopes poor in mineral content (Hadianet al., 2011). It was also observed that the distribution ofS. khuzestanica is very close to Cyclotrichium depauperatum(Bunge) Manden and Sheng (Jamzad, 1994).

Recently, the morphology of glandular trichomes onfresh vegetative and reproductive organs of S. khuzestanica

was investigated and results revealed the presence of one typeof uniseriate, multicellular protective trichome and fourtypes of glandular trichomes including peltate, short-stalkedcapitate, long- stalked capitate, and conoidal trichomes(Dousti et al., 2009).

Since there is no distinct reference concerning therequired parameters for quality control of S. khuzestanicadried leaf as a herbal substance for human use, the presentstudy was carried out to establish the botanical identificationof it according to the official quality monograph specifica-tions for botanical identification of herbal substances, whichincludes the observation and botanical characterization ofthe dried whole, fragmented, and powdered plant material(EDQM, 2010; Upton et al., 2011).

MATERIALS AND METHODS

Plant MaterialMaterial was collected from Lorestan Province in Iran duringfull flowering stage, dried in shade at room temperature, andidentified by Dr. Yousef Ajani. A voucher specimen withnumber 288-ACECR was deposited at the Herbarium of theInstitute of Medicinal Plants, Jahade-Daneshgahi (ACECR),Karaj, Iran. Sixty samples were randomly selected from 250 gof the collected raw material according to the standard

Figure 1. S. khuzestanica leaf macroscopic characters.Adaxial surface view showing: (a) the entire margin, ovate shape,acute apex, attenuate base and ciliate margin; (b, c) peltate glandular trichomes (arrow) and non-glandular trichomes(arrowhead). Scale bars: a – 2000 μm; b, c – 500 μm.

1426 Maryam Malmir et al.

methods of sampling described in European Pharmacopeiafor Herbal Drugs (EDQM, 2010).

Macroscopic AnalysisThe selected samples were examined macroscopicallyaccording to the standard methods described in European

Pharmacopoeia (EDQM, 2010). Shape, size, color, surfacetexture, and fracture type were the main characteristicsobserved. Samples were directly examined by the naked eyeand then by using an Olympus SZ61 stereo microscope(Heerbrugg, Switzerland) coupled with an Olympus Color-View IIIu camera (Tokyo, Japan).

Table 1. Morphological and anatomical characteristics of S. khuzestanica leaf.

Anatomical characteristic Min–Max Mean Median ±SD

Leaf surfaceLength (mm) 5.2–16.3 10.8 10.9 2.6Width (mm) 2.4–8.2 5.8 5.5 1.5

Non-glandular trichomesLength (μm)Type 1 92.3–451.7 237 225.9 89.6Type 2 192.5–337.6 272.6 265.9 52

Glandular trichomesPeltate trichomesLength (μm) 152.6–171.6 163.1 165.1 9.7Head diameter (μm) 98.9–127.3 112.4 113.6 7.7Head area (μm2) 6771–9958 8728 9048 1155

Capitate trichomesType 1 (1SC)Length (μm) 32.4–66.8 41.8 40.2 6.7Head diameter (μm) 10.3–16.8 14.4 14.7 1.5

Type 1 (2SCs)Length (μm) 56.1–84.4 75.9 80 9.9Head diameter (μm) 11–16.2 14.2 14.3 2

Type 1 (3SCs)Length (μm) 93.6–135.7 108.7 105.5 11.7Head diameter (μm) 10–16.3 12.7 12.2 2

Type 1 (4SCs)Length (μm) 142.3–147.4 144.4 143.6 2.6Head diameter (μm) 11.3–13.2 12.1 12 0.8

Type 2Length (μm) 25.4–46.9 36.8 37.2 5.6Head diameter (μm) 16.1–22.8 19.7 19.3 2

StomataAdaxial StomataLength (μm) 12.8–25.3 20.1 20.3 2.6Width (μm) 8.6–18.9 15 15.1 2

Abaxial Stomata (μm)Length (μm) 13.4–25.4 19.1 19 2.5Width (μm) 10.3–17.4 13.7 13.7 1.7

Cross-section featuresLamina thickness (μm) 163.5–313.9 226.5 220.5 46.3Mesophyll thickness (μm) 168.7–247.3 195 188.5 26.9Midrib thickness (μm) 221.3–450.6 340 338 63.8Cuticle thickness (μm) 1–1.6 1.3 1.3 0.2Palisade cell length (μm) 30.1–59.3 41.2 40.4 6.7Spongy cell length (μm) 16.2–31.7 24.4 24.7 4.1Xylem vessel diameter (μm) 9.1–11.1 10.2 10.5 0.7Sclerenchymatous cell diameter (μm) 17.5–56.1 25.4 24.1 9.2Calcium oxalate crystalsCluster crystal diameter (μm) 5–10.3 7.4 7.1 1.3Prismatic crystal width (μm) 7.6–17.7 11 10.6 2.9Raphids crystal length (μm) 4.9–33.5 17.8 17.5 8.7

Min, minimum; Max, maximum; SD, standard deviation; SC, stalk cell.

Botanical Identification of Satureja Khuzestanica Leaf 1427

Light Microscopy (LM)Surface preparations and transverse sections of lamina andmidrib region of the 30 samples of the selected plant materialwere prepared manually. Sample sections were mounted in60% aqueous chloral hydrate solution and examined usingan Olympus CX40 upright microscope (York, UK), coupledwith an Olympus ColorView IIIu camera (Tokyo, Japan).Powdered plant material of 15 samples was obtained usingan Analytical Mill A-10 water-cooled laboratory mill (Staufen,Germany).

Scanning Electron Microscopy (SEM)Fifteen samples of the selected plant material were sectioned,dehydrated at 35°C for 24 h, and directly mounted on stubsusing double-sided adhesive tape. Prepared samples werethen sputtered with a thin layer of gold in a Polaron E 5350and observed using a JEOL JSM-T220 scanning electronmicroscope at 15 kV, with a digital image acquisition inte-grated system (Massachusetts, USA).

Quantitative and Statistical AnalysisQuantification of selected morphological and anatomicalcharacteristics were performed using Olympus Cell^D 2006software. Statistical values were calculated using MicrosoftExcel 2010 software. Stomatal index (SI) was determinedby the formula SI ¼ S ´ 100

S +E where (S) represents the numberof stomata in a given area of the leaf and (E) the number ofepidermal cells (including trichomes) in the same area of theleaf (EDQM, 2010).

RESULTS

Macroscopically (Fig. 1, Table 1), the leaves exhibited agrayish-green color with a characteristic strong and pleasantodor and aromatic taste. They were flat, broadly ovateor orbicular, thin, and fragile, 5–16 mm long and 2–8 mm

wide. The apex was acute, the base was attenuate and themargin ciliate. Venation was pinnate and prominent on theabaxial surface.

Stereomicroscopy examination of both adaxial (upper)and abaxial (lower) leaf surfaces revealed the presence of anindumentum containing bright yellowish points (arrow,Figs. 1b, 1c) and white covering hairs (arrowhead, Fig. 1c),corresponding to glandular and non-glandular trichomes,respectively. Details of the anatomy of such trichomes arepresented in Table 1.

Microscopically, the non-glandular trichomes (Figs. 2a,2b and Table 1) were unicellular to multicellular, composedof one basal epidermal cell, uniseriate, and unbranched. Theywere quite variable in length and could be divided into twotypes: Type I-unicellular to multicellular (curved or straight;arrow, Fig. 2b) acicular trichomes in a single order, and90–450 μm long; and Type II- multicellular trichomeswith ridges and marked internodes and 190–350 μm long(arrowheads, Fig. 2b).

LM and SEM observations of the leaf showed glandulartrichomes, including peltate (Fig. 3 and Table 1) and capitatetypes (Fig. 4 and Table 1), embedded in the surfaces of bothepidermal layers.

The peltate trichomes (Figs. 3a–3h), yellow to orange incolor, consisted of five lengthy stalk cells (multiseriate stalk)(Figs. 3g, 3h) and an enlarged round (smooth or wrinkled)secretory head (Figs. 3a, 3b, 3c, 3f) composed of 12 cells, ofwhich four were small and occupying the central area of thehead, and eight were large and peripheral (Figs. 3a, 3e). A largesubcuticular space in which the secreted material accumulated(subcuticular oil storage cavity) was observed (Figs. 3a, 3c).Raphides (needle-shaped calcium oxalate crystals) were alsoobserved inside the heads of peltate trichomes and also on theadaxial surface of the leaf (Figs. 3c, 3d).

The capitate trichomes (Figs. 4a–4e and Table 1) werequite variable in stalk length and head shape (Table 1). Theywere comprised of one basal cell, a short unicellular or long

Figure 2. (a) SEM and (b) LM photographs of non-glandular trichomes of S. khuzestanica leaf. (a) adaxial surface generalview; (b) details of type I (arrow) and type II (arrowhead) non-glandular trichomes. Scale bars: a – 100 μm, b – 50 μm.


Figure 3. (a, c, e, g) LM and (b, d, f, h) SEM photographs of peltate glandular trichomes of S. khuzestanica leaf. (a) sub-cuticular oil storage cavity and lateral view of the secretory cells (arrow); (b) multiseriate stalk cell (arrow) and smooth headcell (arrowhead); (c) peltate trichome containing the raphids crystals (arrow); (d) glandular trichomes apparently withoutsecretory pores but showing rupture of cuticle (arrows) and raphids crystals of calcium oxalate (arrowhead); (e) top view ofthe central cell (arrow) and secretory cells (arrowhead); (f) wrinkled surface of the peltate trichome head cell (arrow); (g, h)cross-section view of multiseriate stalk cells (arrow). Scale bars: a, b, c, g – 50 μm; e, f, h – 20 μm; d – 10 μm.


multicellular stalk, and one head cell forming a rounded topear shaped secretory head. They were more numerous thanpeltate trichomes and located on both surfaces of the leaf.Among these trichomes, two types could be distinguished.Type I had a short uni- or long multicellular (two to fourcells) stalk and one head cell (Figs. 4a, 4c, 4d, 4e). The cuticlewas thin and there was no subcuticular chamber. Type IIwas very small and had a unicellular stalk and an oblongcutinized secretory head containing a small subcuticularchamber (Fig. 4b).

LM and SEM examination of both adaxial and abaxialleaf surfaces (Figs. 5a–5d and Table 1) showed that thestomata were diacytic and accompanied by two unequalsubsidiary cells (Figs. 5a, 5b). The maximum length of sto-mata was 25 μm and they occurred on both leaf epidermal

surfaces (amphistomatic leaf). They were oval-shaped andlocated a little higher than the epidermal level (hygromorphtype). A striated cuticle was also noticed on the adaxial surface(Figs. 5c, 5d). The adaxial stomatal index, which was defined onthe basis of the number of stomata and number of epidermalcells according to the stomatal index formula, was 13.54.

Examination of the leaf transverse section (Figs. 6a–6dand Table 1) showed that the S. khuzestanica leaf was iso-bilateral with lamina thickness ranging from 160 to 310 μm.The epidermis comprised thick compressed cells. The laminahad differentiated adaxial and abaxial epidermis with a oneto two layered palisade parenchyma and spongy parenchymacells between them (Figs. 6a, 6b). The palisade parenchymacells were cylindrical and the spongy parenchyma cells werecircular or ovoid in transverse section. Collateral vascular

Figure 4. LM photographs of capitate glandular trichomes of S. khuzestanica leaf. (a) type I capitate trichome;(b) type II capitate trichome; (c) capitate trichome type I composed of two stalk cells; (d) capitate trichome type Icomposed of three stalk cells; (e) capitate trichome type I composed of four stalk cells. Scale bars: a–e: 20 μm.


Figure 5. (a, b) LM and (c, d) SEM photographs of stomata type of S. khuzestanica leaf. (a) diacytic stomata (arrow) onadaxial epidermis; (b) abaxial stomata (arrow) with basal cells of trichomes (arrowhead) surrounded by epidermal cells;(c, d) details of the adaxial surface showing the stomata hygromorphic type. Scale bars: a, b – 50 μm; c – 10 μm; d – 5 μm.

Figure 6. (a) SEM and (b, c, d) LM photographs of the cross-section of S. khuzestanica leaf. (a) lamina; (b) detailsof circular to ovoid spongy parenchyma cells together with cluster crystals of calcium oxalate (arrow); (c) midrib;(d) details of a collateral vascular bundle consisting of xylem (arrow) and three layers of sclerenchymatous tissue(arrowhead) near the phloem. Scale bars: a, d – 50 μm; b – 25 μm; c – 100 μm.


bundles, consisting of phloem and xylem and three layersof sclerenchymatous cells, were also located in the centralmesophyll. The midrib in transverse section was concave(Figs. 6c, 6d). Parenchyma cells contained cluster and pris-matic calcium oxalate crystals (Fig. 6b).

LM examination of the powdered leaf (Fig. 7) revealedthe presence of fragments containing all of the above-namedstructures from both adaxial and abaxial epidermal surfaces.The main components were non-glandular trichomes intactor reduced to fragments (Fig. 7a), secretory cells and stalkcells of the peltate trichomes (Figs. 7b, 7c), head cells of type Iand type II capitate trichomes (Figs. 7d, 7e), fragments of themesophyll consisting of palisade parenchyma cells (Fig. 7g),

diacytic stomata (Fig. 7h) and calcium oxalate crystals(Fig. 7i). Hexacolpate, radially symmetrical, oblate-spheroidaland isopolar pollen grains (Fig. 7f) were also observed duringanalysis of the powdered samples.

DISCUSSION

Here we describe how macroscopic and microscopic evalua-tion can be used as a quality assessment tool for the char-acterization of whole, fragmented and powdered samplesof S. khuzestanica leaves. Our macro measurements ofS. khuzestanica dried leaves (length and width), shown in

Figure 7. LM photographs of the powdered leaf of S. khuzestanica. (a) non-glandular trichomes; (b, c) details of secre-tory head cells and stalk cells of the peltate trichome; (d, e) details of head cells of capitate trichomes type I and type II;(f) pollen grain; (g) fragments of the mesophyll, with palisade parenchyma; (h) details of diacytic stomata surroundedby epidermal cells; (i) detail of the vascular bundle, surrounded by a crystalliferous sheath bearing prisms of calciumoxalate (arrow). Scale bars: a – 100 μm; e, f, g, h, i, – 50 μm; b, c, d – 25 μm.


Table 1, are similar to those previously described by Jamzad(1994) and Hadian et al. (2011), but different from themeasurements reported by Kasyani Aval et al. (2012).Micromorphological analyses of Satureja species have beendescribed in several publications (Satil et al., 2002, 2003;Redžić et al., 2006; Dunkić et al., 2007; Satil & Kaya, 2007;Marin et al., 2010, 2012). In all cases, Satureja specieshave non-glandular trichomes together with two types ofglandular trichomes, classed as peltate and capitate, based onmorphological characteristics (Fahn, 2000).

Morphological characteristics of glandular trichomes onfresh vegetative and reproductive organs of S. khuzestanicaand micro-morphology of the fruit and pollen grains werealso previously investigated (Dousti et al., 2009; Moalemet al., 2011). According to our findings, the morphology ofnon-glandular trichomes on dried leaves of S. khuzestanicado not entirely agree with those of Dousti et al. (2009) whodescribed only one type of uniseriate, multicellular protectivetrichome consisting of three to five cells. However, in thissurvey we found some variation in morphology of these tri-chomes and therefore sub-divided them into two differenttypes similar to the findings of Kahraman et al. (2010) usingSalvia chrysophylla Stapf.

In a comparative study of leaf anatomy and trichomesof the genus Satureja in Turkey by Satil and Kaya (2007)the peltate trichomes were found to be composed of onebasal cell, one stalk cell, and a broad 12–16-celled head, 8–12of which were described as large and peripheral, and four ofwhich were said to be small and to occupy the centralarea of the head. According to Dousti et al. (2009), the peltatetrichomes of S. khuzestanica comprised 6–8 voluminous-lengthy stalk cells and 6–8 secretory cells. However, weobserved, on average, 5 voluminous-lengthy stalk cells and12 secretory cells.

As shown in Figures 3b and 3f, the head surface of thepeltate trichomes appeared either smooth or wrinkledand, according to Bosabalidis (1990), the wrinkled surfaceof the peltate trichome head indicates that the cuticle isstill closely attached to the secretory cells, emphasizingthe cell outlines. Moreover, several bundles of needle-likestructures were also observed in our samples, which conformto published characteristics of calcium oxalate raphides(Ryding, 2010).

Dousti et al. (2009) divided the capitate trichomes ofS. khuzestanica into two types according to the dimensionsof the stalk and morphology of the glandular head. However,we observed two types of capitate trichomes, either withor without a subcuticular chamber of the head and fourdifferent subtypes according to the number of stalk cells.Beside the peltate and capitate glandular trichomes, Doustiet al. (2009) also observed conoidal trichomes; however, wedid not detect this type of trichome.

In most herbaceous plants stomatal density is usuallyhigher on the abaxial surface than on the adaxial leaf surface(Willmer & Fricker, 1996). However, we detected no sig-nificant differences between adaxial and abaxial stomataldensity. In an extensive survey of stomatal position in 127

genera of Lamiaceae, Cantino (1990) remarked that bothhypostomatic and amphistomatic leaves are found in thisfamily, the latter type being slightly more frequent. Thestomata in all Satureja species are of hygromorph type orsometimes of hygromorph to mesomorph type. The genusSatureja has equifacial leaves except for S. spicigera, whichhas bifacial leaves (Satil & Kaya, 2007). S. khuzestanicaexhibits amphistomatic, isobilateral leaves with diacytic,hygromorph stomata which are common in the Lamiaceae.

The presence of sclerenchymatous tissue in the vascularbundles, together with cuticle thickness characteristics, areconsidered useful diagnostics in taxonomic studies of thegenus Satureja (Satil & Kaya, 2007). We also documented thepresence of cluster, prismatic and raphide crystals of calciumoxalate in the leaves.

CONCLUSIONS

In conclusion, the pharmacognostic parameters reported inthis study should be included in a quality control monographfor identification and assay purposes of this herbal substance.

ACKNOWLEDGMENT

The authors wish to thank Dr. Yousef Ajani and Dr. ZibaJamzad (Research Institute of Forests and Rangelands,Tehran, Iran) for their valuable assistance in botanicalidentification. They also thank Telmo Nunes from theMicroscopy and Image Analysis (Laboratory of the Centrefor Environmental Biology, Faculty of Sciences, Universityof Lisbon, Portugal), for technical assistance with the SEM.This work was financially supported by FCT–PEst–OE/SAU/UI4013/2011.

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Characterization of Satureja khuzestanica Leaf as a Herbal Medicine