Folk medicines are gaining great importance as information sources on traditional medicinal plants. The aim of this paper is the study of a plant traditionally employed by the Puna inhabitants: Werneria poposa Phil. Morphology of its secretory tissue and other histological diagnostic features, as well as the chemical composition of its essential oil, is described. Puna inhabitants use W. poposa mainly as an infusion for mountain sickness ('soroche'), stomach and hepatic disorders and cold. It is also used externally as a hot bath or unguent for rheumatic pains or traumas. Secretory tissues are schizogenous ducts in leaves and stems. Thirty-one components were detected in the essential oil by means of GC/MS analysis. Oil was characterized by the presence of high content of β-pinene (21.7%), α-pinene (5.5%), terpinen-4-ol (5.3%), α-terpinene (5.2%), β-phellandrene + 1,8-cineole (4.8%), isopulegol (4.8%) and β-citronellal (4.6%). At the present time, W. poposa is consumed as a medicinal plant, mainly in the northwest of Argentina, not being commercialized in the urban centers.Facultad de Ciencias Naturales y Muse

Abella García, Luis

Cortella, Alicia Rita

Pérez Alonso, María J.

Velasco Negueraela, Arturo

Pharmaceutical Biology

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ABSTRACTFolk medicines are gaining great importance as informa-tion sources on traditional medicinal plants. The aim ofthis paper is the study of a plant traditionally employed bythe Puna inhabitants: Werneria poposa Phil. Morphologyof its secretory tissue and other histological diagnosticfeatures, as well as the chemical composition of its essen-tial oil, is described. Puna inhabitants use W. poposamainly as an infusion for mountain sickness (“soroche”),stomach and hepatic disorders and cold. It is also usedexternally as a hot bath or unguent for rheumatic pains ortraumas. Secretory tissues are schizogenous ducts inleaves and stems. Thirty-one components were detected inthe essential oil by means of GC/MS analysis. Oil wascharacterized by the presence of high content of b-pinene(21.7%), a-pinene (5.5%), terpinen-4-ol (5.3%), a-ter-pinene (5.2%), b-phellandrene 1 1,8-cineole (4.8%),isopulegol (4.8%) and b-citronellal (4.6%). At the presenttime, W. poposa is consumed as a medicinal plant, mainlyin the northwest of Argentina, not being commercialized inthe urban centers.INTRODUCTIONEssential oils are responsible for the characteristicscent, odor and flavour of many plants, and are eco-nomically important as the basis of natural perfumes,spices and by their extensive use as medicinal plants.The vast majority of these drugs are used in crude form,mainly for the preparation of infusions (Bruneton,1995). Many aromatic plants are used by the inhabi-tants of Puna as therapeutical resources; they can rec-ognize them by odor and some morphological features.Folk medicines are gaining importance as a sourceof information for traditional medicinal plants(Hostettmann et al., 1995). These drugs are subjected toinvestigation in the search for new biodynamic com-pounds with renewed rigor. A common belief is thatplant remedies are naturally superior to synthetic drugsand that they are not harmful to human beings (Beier,1992).The goal of this paper is to establish traditional andcurrent uses of W. poposa by the Puna inhabitants, themorphology of secretory tissue and other histologicaldiagnostic features, as well as the chemical composi-tion of its essential oil.Werneria poposa Phil. (common names: poposa,pupusa [Compositae]) has the following characteris-tics: fetid, very branchy, cushion forming, sub-shrub,(cojín), 3–5 cm height, stems densely leafy; wide innerwoolly leaf sheath. Apical solitary sessile and radialcapitulum, campaniform involucral with glabrousoblong phyllary. Dimorphic tubulose hermaphroditicflowers; white ligulate radial flowers and yellow-lili-aceus disc flowers. Cylindricous glabrous ribbed ach-ene, white-yellow papus (Fig. 1). It grows in highmountains of South America: South Perú, North Chileand North West Argentina (from Jujuy to Tucuman).4600–5300 m above sea level. This region is known asPuna (Cabrera, 1978) (Fig. 2).MATERIALS AND METHODSThe above ground parts of the plants were collected inPuna with the help of local informants, and the materialPharmaceutical Biology 1388-0209/00/3803-0197$15.002000, Vol. 38, No. 3, pp. 197–203 © Swets & ZeitlingerETHNOBOTANY, VOLATILE OILS AND SECRETION TISSUES OFWERNERIA POPOSA FROM ARGENTINALuis Abella1, Alicia R. Cortella1*, Arturo Velasco-Negueruela2 and María J. Pérez-Alonso21LEBA, Fac.Cs.Nat. y Museo, Univ. Nacional de La Plata, Paseo del Bosque s/n, 1900–La Plata, Argentina2Departamento de Biología Vegetal I, Universidad Complutense de Madrid, 28040–Madrid, EspañaKeywords: Werneria poposa, Puna, ethnomedicine, essentialoils, secretory tissues.Address correspondence to: Alicia R. Cortella, Laboratorio deEtnobotánica y Botánica Aplicada (LEBA), Facultad de Cien-cias Naturales y Museo, Universidad Nacional de La Plata,Paseo del Bosque S/N, 1900, La Plata, Argentina. Fax: 54-0221-425-7527, E-mail: rcastell@dalton.quimica.unlp.edu.arwas also purchased at local markets. The plant wasbotanically confirmed using available literature (Cabr-era, 1978).A voucher specimen was deposited at the Museo deLa Plata herbarium and at the Laboratorio de Etnob-otanica y Botánica Aplicada (LEBA) herbarium No. 9,L. Abella.The area was surveyed four times between 1995 and1997, in different seasons, to perform ethnobotanicalstudies. Data were obtained from native informantswho were healers, faith healers, ordinary villagers andmarket traders. The information was collected bymeans of direct observation, participating observationinterviews and tape recordings.Samples were hydrated in water during 24 h for his-tological studies, performed with a light microscope(LM). Sections were performed with frozen microtomeMicrom HM 340 E and also with a manual one. Oxi-dant agents H2O2 (100 vol) and NaClO were used toclear the sections. In order to detect terpenoids histo-chemically, the SbCl3/HClO4 reagent, that generates ared colour, was used (Mace et al., 1976).Scanning electron microscope (SEM) was also used.Fragments of the leaves and stems were Critical Pointdried. The samples were previously fixed with FAA(formaline 1 anhydrous acetic acid 1 anhydrousethanol) for four days, and then successively washed inethanol, 50, 70 and 100%.The secretion tissue was described; in addition,other histological characters were studied which couldbe useful as diagnostic features to identify fragmented,powdered or destroyed samples.Air-dried plant material was hydrodistilled in an allglass Clevenger-type apparatus to isolate volatile con-stituents. The oil was dried over anhydrous sodium sul-phate and stored at 4°C in the dark.Analytical GC was performed on a Varian 3300 gaschromatograph equipped with a silicone DB-1 capillarycolumn (30 m 3 0.25 mm, film thickness 0.25 mm)using nitrogen at a 1.5 ml/min flow rate as carrier gas.Injections were made by the split mode, and columntemperature programmed from 80 to 225°C at 4°C/min.Injector temperature was 250°C, and a FID detector at300°C was used. Injection volume was 0.1 ml for allsamples. GC/MS analysis was carried out on a HewlettPackard 5890 gas chromatograph equipped with a sili-cone SE-30 capillary column (20 m 3 0.2 mm) at a car-rier gas flow rate of 1.5 ml/min. Temperature wasprogrammed from 70 to 210°C at 4°C/min and theinjector temperature was 250°C. The chromatographwas coupled to a HP 5971 A mass selective detector(70 eV). All constituents were identified by comparingtheir Kováts retention indices with those of authenticstandards available in the author’s laboratory. The lat-ter were compared with those stored in the spectrome-ter database using the commercial NBS54K.L andWILEY.L built-in libraries and with those reported inthe literature (Libey, 1991; Adams, 1995).RESULTS AND DISCUSSIONW. poposa is widely used as a therapeutic resource inPuna. The most common method of preparation is asinfusion: less than a handful of leaves and stems in a198 L. ABELLA ET AL.Fig. 2. Puna geographical location.Fig. 1. W. poposa. Plant aerial parts.cup of tea (250 ml of boiling water) are used for“soroche” (mountain sickness). It is also used as adigestive and for hepatic colics, cough, cold and bron-chitis, and as a food condiment for pneumonia conva-lescents. Former reports dealing with some of theseuses in the North West of Argentina (Palma, 1973;Perez De Nucci, 1988; Martinez & Pochettino, 1992)were confirmed in the present study.External use is also very common, as a hot bath,mainly for the feet, or associated with Artemisia copa(“copa-copa”) for rheumatic pains (Palma, 1973).Bibliographic references mention its use in thepreparation of unguents (powdered plant mixed withhen fat) to treat rheumatic and traumatic pains, and alsoas a local antiinflamatory (Meyer, 1943; Zardini, 1984).The use of the root in pleurisy has been reported in onecase (Debelmas, 1975).At the present time, W. poposa is consumed as amedicinal plant, mainly in the northwest of Argentina,not being commercialized in urban centers.Secretory tissues are schizogenous ducts which gothrough leaves and stems (Figs. 3, 4, 5); their diameterAROMATIC PLANTS FROM PUNA 199Fig. 3. Leaf schizogenous duct critical point dry.Fig. 4. Cross section leaf (LM).is uniform (350–400 mm) and they are internally coatedwith thin-walled epithelial secretory cells (Fig. 6). Pos-itive terpenoid histochemical reactions were detected inall the tests performed on these tissues or in secretionproducts.Volatile oils are pressurized within the ducts, and aslight traumatism produced in the plant immediatelyreleases the essential oil. Poposa is a very aromaticsmelling plant.In addition to the secretory tissues, the main histo-logical diagnostic features are the abundant whip-liketrichomes present in the leaf sheath, with two basalcells and a very long apical (1–1.5 mm), twisted, thickwall cell (Fig. 7). The leaf sheath is very woolly; theseleaf sheath hairs are characteristic of other Compositaewhose apical cells are not as long and thick as in W.poposa (Metcalfe & Chalk, 1979).Other features are the deep stomata and a thick waxylayer of the aerial parts, characteristic of xerophyticplants (Fig. 8) (Ancibor, 1980).The components of the oil, the percentage by weightof each constituent and their Kováts indices are200 L. ABELLA ET AL.Fig. 5. Stem schizogenous duct. Essential oil filling in the cavity (arrow).Fig. 6. Epithelial secretory cells (LM).summarized in Table 1. The components are arrangedin the order of GC elution from the silicone columns.The yield of essential oil was about 1% based on driedweight.The oil was characterized by the presence of a highcontent of b-pinene (21.8%); other important compo-nents were found to be a-pinene (5.5%), terpinen-4-ol(5.3%), a-terpinene (5.2%), b-phellandrene 1 1.8- cine-ole (4.8%), isopulegol (4.8%) and b-citronellal (4.6%).The a- and b-pinenes produce an increase of bloodflow in the skin and in the mucous membranes; thisexplains its external use for rheumatic pains and asinfusion for digestive problems. This monoterpene isalso a well know antiseptic (Duke, 1991). Terpenes arefound in the leaves of some desert plants and are thecause of allelopathic effects (Harbone, 1977).There are no records about intoxications caused bythe use of poposa, though it is widely consumed bypeople. Evidently the Puna inhabitants know about thedose to be used for various health problems.AROMATIC PLANTS FROM PUNA 201Fig. 7. Leaf sheath trichomes.Fig. 8. Leaf upper surface, deep stomate and waxy layer.ACKNOWLEDGEMENTSFinancial support from the Consejo Nacional de Investiga-ciones Científicas y Técnicas (CONICET), UniversidadNacional de La Plata and Agencia Española de CooperaciónInternacional, Ministerio de Asuntos Exteriores de Españais gratefully acknowledged. The authors would like to thankthe technicians of the SEM of the Museo Nacional de LaPlata.REFERENCESAdams RP (1995): Identification of Essential Oils Compo-nents by Gas Chromatography Mass Spectroscopy, Illi-nois, II, Allured Publishing Co.Ancibor E (1980): Estudio anatómico de la vegeteción de lapuna de Jujuy II, Anatomía de las plantas en cojín. BolSoc Argen Bot XIX 1–2: 157–202.Beier RC (1992): Natural toxicants in foods. In: Nigg HN, Sei-gler D, eds., Phytochemical Resources for Medicine andAgriculture, pp. 247–367, New York, London, PlenumPress.Bruneton J (1993): Pharmacognosy, Phytochemistry, Medic-inal Plants, pp. 425–426, Paris Lavoisier Publishing.Cabrera AL (1978): Flora de la provincia de Jujuy (Com-positae), Col Ci INTA 13(10): p. 474.Debelmas J (1975): Plantas medicinales d’altitude. Fitoter-apia 3: 99–110.Duke JA (1991): Handbook of Medicinal Herbs, USA, CRCPress Inc. Harbone JB (1977): Introduction to Ecological Biochemistry,pp. 185–187, London, Academic Press, Hostettmann K, Marstons A, Maillard M, Hamburger M(1995): Phytochemistry of Plant Used in Traditional Med-icine. Proceedings of the Phytochemical Society ofEurope, Oxford, Claredon Press.Libey LM (1991): A paradox data base for GC/MS data oncomponents of essential oils and other volatiles. J EssentOil Res 3: 192–1942.Mace ME, Bell AA, Beckman CH (1976): Histochemistryand identification of disease-induced terpenoid aldehydesin Verticillium-wilt-resistant and susceptible cottons. CanJ Bot 54: 2095–2099.Martinez MR, Pochettino ML (1992): The “farmacia casera”(household pharmacy): A source of ethnopharmaco-botanical information. Fitoterapia 3: 209–217.202 L. ABELLA ET AL.Table 1. Major components found in W. poposa essential oil.Component % tR I1.- a-thujene 1.9 3.4 9282.- a-pinene 5.5 3.4 9283.- camphene 0.4 3.6 9414.- sabinene 2.0 3.9 9685.- b-pinene 21.8 3.9 9686.- myrcene 0.3 4.2 9847.- a-phellandrene 0.5 4.3 9958.- a-terpinene 5.2 4.5 10059.- p-cymene 2.4 4.7 101710.- b-phellandrene11,8-cineole 4.8 4.7 101711.- cis-ocimene 1.2 5.1 103312.- Isopenthyl-n-butirate 1.2 5.2 103913.- g-terpinene 1.8 5.2 104114.- cis-linalool oxide 0.2 5.5 105415.- terpinolene 0.7 5.8 107116.- linalool 2.2 5.9 107817.- isopulegol 4.8 7.0 112318.- citronellal 1.3 7.3 113419.- iso-isopulegol 0.5 7.4 114120.- terpinen-4–ol 5.3 7.7 115221.- a-terpineol 2.1 8.2 116422.- b-citronellol 4.6 9.0 120223.- neral 2.6 9.9 122224.- geraniol 0.3 9.9 123525.- geranial 2.8 10.1 125026.- a-copaene 0.2 13.6 136327.- aromadendrene t 15.6 142528.- g-muurolene 0.3 16.5 145629.- a-muurolene 0.1 17.1 147830.- g-cadinene 0.8 17.6 149331.- d- cadinene 2.9 17.9 150132.- b-eudesmol 1.9 21.5 1623%: percentage by weight percentage in the oil mixture tR : retention time (min). I: Kováts retention index. (t): components found as traces(, 0.1%).Metcalfe CR, Chalk L (1979): Anatomy of the Dicotyledons,Vol I, p. 192, Oxford, Clarendon Press. Meyer T (1943): Notas sobre un viaje etnobotánico aldepartemento de Santa Victoria (Salta), Rev Geogr Amer10: 121–132.Palma NH (1973): Estudio antropológico de la medicina pop-ular de la Puna argentina, p. 100, Argentina, Carbagó.Pérez De Nucci A (1988): La medicina tradicional delnoroeste argentino. Historia y presente, Serie Antropológ-ica, p. 131, Ed. del Sol, Buenos Aires, Argentina. Zardini EM (1984): Etnobotánica de compuestas argentinas,con especial referencia a su uso farmacológico, ActaFarma Bonaerense 3(1): 77.Accepted: February 22, 2000AROMATIC PLANTS FROM PUNA 203

Ethnobotany, volatile iols and secretion tissues of Werneria poposa from Argentina

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Ethnobotany, volatile iols and secretion tissues of Werneria poposa from Argentina

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