Chemical Analysis of the Essential Oils from Three Populations of Lippia dulcis Trevir. Grown at Different Locations in Southern Ecuador.

In this investigation, we have analyzed for the first time the essential oils (EOs) isolated by steam distillation of the leaves and flowers of Lippia dulcis Trevir., grown at three different locations in southern Ecuador: the Catacocha canton (Ca), the Vilcabamba parish (Vi), and the Chuquiribamba parish (Ch). Around 98.5% of the oils' constituents were identified by Gas Chromatography-Mass Spectrometry (GC-MS) and Gas Chromatography-Flame Ionization Detector (GC-FID) analysis using a DB-5ms capillary column. Sesquiterpene hydrocarbons were predominant in the EOs (79.77, 78.22, and 76.51%, respectively). The most representative constituents of the sample from the Ca canton were ß-cedrene (16.75%), δ-selinene (11.04%), and ß-cubebene (12.09%), while the sample from the Vi parish was characterized by the abundant presence of ß-cedrene (17.9%), δ-selinene (12.52%), and bicyclogermacrene (11.34%). ß-Cedrene (18.89%), δ-selinene (11.78%), and δ-cadinene (11.07%) were the main constituents of the essential oil (EO) from the Ch parish. The likely occurrence of low amounts of thermolabile hernandulcin in the volatile oils was indicated by the presence of the fragmentation products 6-methyl-5-hepten-2-one and 3-methyl-2-ciclohexen-1-one. In summary, the study gave us a clue to the variability of Lippia dulcis chemotypes depending on the collection sites.

New Tricholomalides D-G from the Mushroom Tricholoma ustaloides Grown in an Italian Beech Wood.

Four novel seconeodolastane diterpenoids, named tricholomalides D-G, were isolated, together with the known tricholomalide C, from the fruiting bodies of Tricholoma ustaloides Romagn., a species belonging to the large Tricholoma genus of higher mushrooms (Basidiomycota, family Tricholomataceae). They were isolated through multiple chromatographic separations, and the structures, including the absolute configuration, were established through a detailed analysis of MS, NMR, and CD spectral data and comparison with related compounds reported in the literature, which has been thoroughly revised.

A Validated HPLC-UV-ESI-IT-MS Method for the Quantification of Carnosol in Lepechinia mutica, a Medicinal Plant Endemic to Ecuador.

The diphenolic diterpene carnosol was isolated from several species of the family Lamiaceae, including Lepechinia mutica, a medicinal plant endemic to Ecuador. The compound has exhibited high antioxidant, anti-inflammatory, antimicrobial, neuroprotective, and antifungal properties, as well as promising cytotoxicity against prostate, breast, skin, leukemia, and human colon cancer cell lines. In this paper, we developed and validated a simple, accurate, and reliable analytical HPLC-UV-ESI-IT-MS method, carried out on a C18 column, which is potentially suitable to quantify carnosol in plant extracts. The procedure complied with the established ICH validation parameters of analytical range (linearity in the range of 0.19-5.64 µg/g dried leaves; REAVERGE = 4.9%; R2 = 0.99907), analysis repeatability (RSD = 2.8-3.6%), intermediate precision (RSD = 1.9-3.6%), accuracy (estimated as % carnosol recovery in the range of 81 to 108%), and robustness. Finally, the LOD (0.04 µg/mg) and LOQ (0.19 µg/mg) values of carnosol/dried leaves were determined. Using this validated method, the content of carnosol in L. mutica was estimated to be 0.81 ± 0.04 mg/g of dried leaves (0.081%).

Constituents and Selective BuChE Inhibitory Activity of the Essential Oil from Hypericum aciculare Kunth.

A potential source of new inhibitors of cholinesterase enzymes are certain compounds of natural plant origin; therefore, in the study described herein we have determined the chemical composition and the acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibitory activities of the essential oil (EO) steam distilled from aerial parts of Hypericum aciculare, which was collected in southern Ecuador. The oil qualitative and quantitative composition was determined by GC-FID and GC-MS using a non-polar and a polar chromatographic column. A total of fifty-three constituents were identified, that accounted for about 98% of the EO content. The hydrocarbon n-nonane (16.4-28.7%) and the aldehyde n-decanal (20.7-23.1%) were the predominant oil constituents. In addition, the EO showed significant inhibition of BuChE (IC50 = 28.3 ± 2.7 µg/mL) and moderate activity towards AChE (IC50 = 82.1 ± 12.1 µg/mL). Thus, the EO from H. aciculare aerial parts is an interesting candidate to investigate the mechanism of selective ChE inhibition by the two ChE enzymes with the aim to discover potential targets to control the progression of the Alzheimer's disease (AD).

New Tricholidic Acid Triterpenoids from the Mushroom Tricholoma ustaloides Collected in an Italian Beech Wood.

The secondary metabolites produced by Tricholoma ustaloides Romagn., a mushroom species belonging to the large Tricholoma genus (Basidiomycota, Tricholomataceae), are unknown. Therefore, encouraged by the interesting results obtained in our previous chemical analyses of a few Tricholoma species collected in Italian woods, we aimed to investigate the secondary metabolites of Tricholoma ustaloides. The chemical analysis involved the isolation and characterization of secondary metabolites through an extensive chromatographic study. The structures of isolated metabolites, including the absolute configuration, were established based on a detailed analysis of MS, NMR spectroscopic, optical rotation, and circular dicroism data, and on comparison with those of related compounds reported in the literature. Two novel lanostane triterpenoids, named tricholidic acids B and C, together with triglycerides, a mixture of free fatty acids, five unidentified metabolites, and the known rare saponaceolides F and J, tricholidic acid, and tricholomenyn C, were isolated from an EtOAc extract of fruiting bodies of Tricholoma ustaloides that were collected in an Italian beech wood. This is the second example of isolation of tricholidic acid derivatives from a natural source. Saponaceolides F and J exhibited high cytotoxicity (IC50 values ≤ 10 µM) against a panel of five human cancer cell lines. The toxicity against myeloid leukemia (HL-60), lung cancer (A-549), hepatocellular cancer (HepG2), renal cancer (Caki-1), and breast cancer (MCF-7) cells was higher than that shown by the very well-known cytotoxic drug cisplatin.

Constituents, Enantiomeric Content, and ChE Inhibitory Activity of the Essential Oil from Hypericum laricifolium Juss. Aerial Parts Collected in Ecuador.

The physical properties, chemical composition, enantiomer distribution, and cholinesterase (ChE) inhibitory activity were determined for a steam-distilled essential oil (EO), with a yield of 0.15 ± 0.05 % (w/w), from H. laricifolium aerial parts, collected in southern Ecuador. The oil qualitative and quantitative analyses were performed by GC-EIMS and GC-FID techniques, using two capillary columns containing a non-polar 5%-phenyl-methylpolysiloxane and a polar polyethylene glycol stationary phase, respectively. The main constituents (>10%) detected on the two columns were, respectively, limonene (24.29, 23.16%), (E)-ß-ocimene (21.89, 27.15%), and (Z)-ß-ocimene (12.88, 16.03%). The EO enantioselective analysis was carried out using a column based on 2,3-diethyl-6-tert-butyldimethylsilyl-ß-cyclodextrin. Two mixtures of chiral monoterpenes were detected containing (1R,5R)-(+)-α-pinene (ee = 83.68%), and (S)-(-)-limonene (ee = 88.30%) as the major enantiomers. This finding led to some hypotheses about the existence in the plant of two enantioselective biosynthetic pathways. Finally, the EO exhibited selective inhibitory effects in vitro against butyrylcholinesterase (BuChE) (IC50 = 36.80 ± 2.40 µg/mL), which were about three times greater than against acetylcholinesterase (IC50 = 106.10 ± 20.20). Thus, the EO from Ecuadorian H. laricifolium is an interesting candidate for investigating the mechanism of the selective inhibition of BuChE and for discovering novel drugs to manage the progression of Alzheimer's disease.

Essential Oil and Non-Volatile Metabolites from Kaunia longipetiolata (Sch.Bip. ex Rusby) R. M. King and H. Rob., an Andean Plant Native to Southern Ecuador.

Kaunia longipetiolata (Sch.Bip. ex Rusby) R. M. King and H. Rob. (Asteraceae) is a plant native to southern Ecuador. The dry leaves afforded, by steam distillation, an essential oil that was qualitatively and quantitatively analyzed by GC-MS and GC-FID, respectively, on two orthogonal columns of different polarity. Sesquiterpenes predominated in the volatile fraction, among which α-zingiberene (19.7-19.1%), ar-curcumene (17.3-18.1%), caryophyllene oxide (5.1-5.3%), (Z)-ß-caryophyllene (3.0-3.1%), (2Z,6Z)-farnesal (2.6-3.6%), and spathulenol (2.0-2.1%) were the major components. In addition to the identified compounds, two main unidentified constituents (possibly oxygenated sesquiterpenes) with probable molecular masses of 292 and 230, respectively, were detected. They constituted about 5% and 8% (w/w), respectively, of the whole essential oil. The oil chemical composition was complemented with the enantioselective analysis of ten chiral components. Four scalemic mixtures and six enantiomerically pure terpenes were identified. An enantiomeric excess (ee) was determined for (1R,5R)-(+)-ß-pinene (65.0%), (R)-(-)-α-phellandrene (94.6%), (S)-(+)-linalool (15.0%), and (R)-(-)-terpinen-4-ol (33.8%). On the other hand, (1R,5R)-(+)-α-pinene, (1R,5R)-(+)-sabinene, (S)-(-)-limonene, (S)-(+)-ß-phellandrene, (1R,2S,6S,7S,8S)-(-)-α-copaene, and (R)-(+)-germacrene D were enantiomerically pure. Finally, the non-volatile fraction obtained by extraction of the leaves with MeOH was investigated. Eight known compounds were isolated by liquid column chromatographic separations. Their structures were determined by NMR spectroscopy as dehydroleucodine, kauniolide, (3S,3aR,4aR,6aS,9aS,9bR)-3-hydroxy-1,4a-dimethyl-7-methylene-5,6,6a,7,9a,9b-hexahydro-3H-oxireno[2',3':8,8a]azuleno[4,5-b]furan-8(4aH)-one, novanin, bisabola-1,10-diene-3,4-trans-diol, (R)-2-(2-(acetoxymethyl)oxiran-2-yl)-5-methylphenyl isobutyrate, eupalitin-3-O-glucoside, and 3,5-di-O-caffeoylquinic acid. Literature data about the identified metabolites indicate that K. longipetiolata is a rich source of biologically active natural products.

Chemical Characterization and Biological Activity of the Essential Oil from Araucaria brasiliensis Collected in Ecuador.

The purpose of this study was to determine the chemical composition, physical properties, enantiomeric composition and cholinesterase inhibitory activity of the essential oil (EO) steam-distilled from the leaves of the plant Araucaria brasiliensis Loud. collected in Ecuador. The chemical composition was determined by gas chromatography coupled to mass spectrometry (GC-MS) analysis on two capillary GC columns (DB5-ms and HP-INNOWax). Thirty-three compounds were identified in the EO; the main compounds were beyerene (26.08%), kaurene (24.86%), myrcene (11.02%), α-pinene (9.99%) and 5,15-rosadiene (5.87%). Diterpene hydrocarbons (65.41%), followed by monoterpene hydrocarbons (21.11%), were the most representative components of the EO. Enantioselective analysis of the EO showed four pairs of enantiomeric compounds, α-pinene, camphene, γ-muurolene and δ-cadinene. In an in vitro assay, the EO showed moderate inhibitory activity towards the enzyme butyrylcholinesterase (BuChE) (95.7 µg/mL), while it was inactive towards acetylcholinesterase (AChE) (225.3 µg/mL). Further in vivo studies are needed to confirm the anticholinesterase potential of the EO.

Poorly Investigated Ecuadorian Medicinal Plants.

Ecuador has, in proportion of its size, one of the richest floras of Latin America and the world; the country also has an immense cultural heritage due to the presence of different ethnic groups that have implemented the use of many wild and cultivated plants, mainly as medicinal remedies. In a recent publication, we have summarized the results of research activities recently carried out on about 120 plants native to Ecuador, which includes the structures of non-volatile isolated compounds, as well as the chemical composition of essential oils (EOs) and the in vitro tested biological activity data. For the sake of completeness, we have collected in this paper the main information obtained from recent ethnobotanical investigations on other important Ecuadorian medicinal plants for which phytochemical, pharmacological, and toxicological studies are, however, still largely lacking. Thus, one of the objectives of this paper is to preserve the traditional knowledge of Ecuadorian Indigenous communities which, being transmitted orally, is in danger of becoming lost. Moreover, it is our intention to stimulate more extensive studies on the rich medicinal flora of the country, which can provide economic and social benefits, especially to the people who traditionally cultivate or collect the plants.

Pharmacology and Phytochemistry of Ecuadorian Medicinal Plants: An Update and Perspectives.

The use of plants as therapeutic agents is part of the traditional medicine that is practiced by many indigenous communities in Ecuador. The aim of this study was to update a review published in 2016 by including the studies that were carried out in the period 2016-July 2021 on about 120 Ecuadorian medicinal plants. Relevant data on raw extracts and isolated secondary metabolites were retrieved from different databases, resulting in 104 references. They included phytochemical and pharmacological studies on several non-volatile compounds, as well as the chemical composition of essential oils (EOs). The tested biological activities are also reported. The potential of Ecuadorian plants as sources of products for practical applications in different fields, as well the perspectives of future investigations, are discussed in the last part of the review.

Essential Oil and Major Non-Volatile Secondary Metabolites from the Leaves of Amazonian Piper subscutatum.

The essential oil and the major non-volatile secondary metabolites from the leaves of Piper subscutatum (Miq.) C. DC. (Family Piperaceae), collected in the Ecuadorian Amazon, were analyzed for the first time in the present study. The essential oil was submitted to chemical and enantioselective analyses by GC-MS and GC-FID. (E)-ß-caryophyllene (25.3-25.2%), ß-chamigrene (10.3-7.8%), (E)-nerolidol (8.1-7.7%), ß-selinene (7.2-7.7%), δ-cadinene (2.7-3.9%), bicyclogermacrene (3.7-2.4%), and ß-pinene (2.6-3.4%) were the major components. The enantioselective analysis, carried out on a ß-cyclodextrin-based column, showed four scalemic mixtures in which (1R,5R)-(+)-α-pinene, (1S,5S)-(-)-ß-pinene, (S)-(-)-limonene, and (1R,2S,6S,7S,8S)-(-)-α-copaene were the major enantiomers, with enantiomeric excesses of 28.8%, 77.8%, 18.4%, and 6.0%, respectively. The study was complemented with the chemical analysis of the organic fraction dissolved in the hydrolate, whose major components were 6-methyl-5-hepten-2-one (63.7-64.4%) and linalool (6.5-6.0%). Concerning the non-volatile fraction, five lignans were the major components. (-)-Beilshminol B, (-)-grandisin, (-)-3',4'-methylenedioxy-3,4,5-trimethoxy-7,7'-epoxylignan, (-)-3',4'-methylenedioxy-3,4,5,5'-tetramethoxy-7,7'-epoxylignan, and (-)-3,4,3',4'-dimethylenedioxy-5,5'-dimethoxy-7,7'-epoxylignan were identified by means of NMR spectroscopy, mass spectrometry and X-ray crystallography. The absolute configuration 7S,8S,7'S,8'S was tentatively assigned to all of them.

Selective BuChE Inhibitory Activity, Chemical Composition, and Enantiomeric Content of the Essential Oil from Salvia leucantha Cav. Collected in Ecuador.

The essential oil (EO) of Salvia leucantha Cav. was isolated by steam distillation of the aerial parts collected in the South of Ecuador. Its physical properties were evaluated and the chemical composition of the oil was determined by GC-MS and GC-FID analyses using two chromatographic columns, DB-5ms and HP-INNOWax. Six major compounds were identified, namely, the sesquiterpenes 6.9-guaiadiene (19.14%), (E)-caryophyllene (16.80%), germacrene D (10.22%), (E)-ß-farnesene (10.00%), and bicyclogermacrene (7.52%), and the monoterpenoid bornyl acetate (14.74%). Furthermore, four pairs of enantiomers were determined by enantioselective GC-MS of the essential oil. (-)-germacrene D and (+)-α-pinene showed the highest enantiomeric excess (ee%). In an in vitro assay, the essential oil demonstrated an interesting inhibitory activity of the enzyme butyrylcholinesterase (BuChE), with an IC50 = 32.60 µg/mL, which is the highest determined for a Salvia species. In contrast, the oil was weakly active against acetylcholinesterase (AChE) with an IC50 > 250 µg/mL.

Chemical composition and selective BuChE inhibitory activity of the essential oils from aromatic plants used to prepare the traditional Ecuadorian beverage horchata lojana.

ETHNOBOTANICAL AND ETHNOMEDICINAL RELEVANCE: In southern Ecuador, horchata lojana is a popular aromatic and refreshing beverage that is prepared from an aqueous infusion of different mixtures of local medicinal and aromatic plants. The drink is considered a traditional anti-inflammatory agent and brain tonic; due these properties, it has been drunk since Colonial Times. Several pharmacological studies have evaluated the effects of horchata aqueous infusion. However, the aromatic profile and the contribution of the volatile components to the biological activity of the drink have not been investigated so far. For these reasons, we have determined the chemical composition of the essential oils (EOs) distilled from five mixtures of aromatic plants commonly used for the preparation of this traditional drink. Moreover, to support the curative properties of the aromatic plants, the anticholinesterase activity of the EOs was examined. MATERIAL AND METHODS: Different bunches of fresh mixed medicinal and aromatic plants, called tongos, are sold at local markets in the province of Loja for the preparation of different types of horchata. In this research we have purchased plant bunches sold at five popular markets of Loja province. Subsequently, aromatic plants in each bunch were separated from medicinal plants and were then hydrodistilled to give the corresponding EOs. Subsequently, the chemical composition of each EO was determined by GC-MS/GC-FID techniques, whereas the cholinesterase inhibitory activity in vitro was determined against acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) enzymes. AIMS OF THE STUDY: i) to contribute to the chemical and pharmacological study of the aroma components of the traditional Ecuadorian drink horchata lojana; ii) to identify botanically the mixtures of aromatic plants used to make the drink; iii) to establish, on the basis of the chemical composition of the EOs, the compounds mainly responsible for the characteristic beverage flavor; iv) to establish the possible existence of an aromatic pattern characteristic of each horchata preparation; v) to test the anticholinesterase activity of the EOs against AChE and BuChE in order to support the traditional consume of the drink as an effective brain tonic. RESULTS: A total of 23 botanical families and 32 species of plants used for the preparation of five different variants of the traditional horchata lojana beverage, have been identified. Fourteen aromatic species were determined to be responsible for the characteristic flavor of the drink. All the analyzed EOs belong to the monoterpene type. A total of 88 compounds have been identified in the different EOs, twenty-four of which are common components of the oils. CONCLUSIONS: According to the main components of the EOs distilled from the five groups of horchata lojana plants, four aromatic profiles have been defined: (i) neral + geranial + carvone, (ii) neral + geranial + myrcene; (iii) geranial + methyl eugenol + isomenthone + neral + citronellol; (iv) (E)-anethole + geranial + pulegone. Moreover, according to the literature, several aromatic plants and individual EOs components exhibit a wide range of biological activities. This finding as well as the significant BuChE inhibitory activity exhibited in vitro by the EOs give scientific support to the use of identified aromatic plants in the traditional preparation of horchata, that is considered a natural analgesic and anti-inflammatory remedy, and an effective brain tonic.

Selective BuChE inhibitory activity, chemical composition, and enantiomer content of the volatile oil from the Ecuadorian plant Clinopodium brownei

ABSTRACT This paper describes the chemical composition and the enantiomer content of the volatile oil hydrodistilled from Clinopodium brownei (Sw.) Kuntze, Lamiaceae. The plant was collected in the South of Ecuador. Thirty one components were identified by GC-MS, which accounted for the 96.15% of the volatile oil. The major components were pulegone (48.44%), menthone (34.55%) and β-acorenol (3.41%). Oxygenated monoterpenes (86.06%), followed by oxygenated sesquiterpenes (5.36%) constituted the most abundant fractions. The enantiomeric compositions of β-pinene, sabinene, 3-octanol, menthone, pulegone and menthyl acetate were determined by enantioselective GC-MS. (-)-Menthone showed the highest enantiomeric excess (ee = 83.4%). In in vitro tests, the volatile oil showed high selective inhibitory activity for butyrylcholinesterase with an IC50, 13.4 ± 1.8 µg/ml. In contrast, it was weakly active against acetylcholinesterase with an IC50 >250 µg/ml.

Carnosol from Lepechinia mutica and tiliroside from Vallea stipularis: Two promising inhibitors of BuChE

Abstract Lepechinia mutica (Benth.) Epling, Lamiaceae, and Vallea stipularis L.f., Elaeocarpaceae, are the object of the present study. These plants are endemic to the Andean region and have attracted our attention on the basis of interesting results obtained in a preliminary anticholinesterase screening. Actually, carnosol and tiliroside, isolated from L. mutica and V. stipularis, respectively, have shown a promising selective inhibitory activity against butyrylcholinesterase. Specifically, the anti-butyrylcholinesterase activity of carnosol was 5.15 µM and that of tiliroside was 52.9 µM, compared to 8.568 ± 0.570 µM of the positive control Donepezil. Carnosol and tiliroside were purified chromatographically from the ethyl acetate extract of L. mutica and V. stipularis, respectively. Spectrophotometric methods were used for enzymatic studies.

Chemical composition and antimicrobial activity of Myrcianthes fragrans essential oil, a natural aromatizer of the traditional Ecuadorian beverage colada morada.

ETHNOBOTANICAL AND ETHNOMEDICINAL RELEVANCE: The importance given in Ecuador to the rescue of traditional knowledge and nutritional value of ancestral foods and drinks, has stimulated our investigation of the chemical composition and some biological activities of M. fragrans ('arrayán') essential oil, a natural aromatic additive used in the preparation of the traditional fruit-juice 'colada morada', which is typically drunk in the Day of the Dead or All Soul´s Day. MATERIAL AND METHODS: Different essential oils of Myrcianthes fragrans (Sw.) McVaught were obtained by hydrodistillation of the aerial parts of the plant collected in Cerro Villonaco (Loja-Ecuador) at three different phenological growth stages, i.e., during foliation (Fo), flowering (Fl) and fruiting (Fr) stages. The chemical compositions of the essential oils were determined by GC/MS and GC/FID techniques. The antimicrobial activities were determined by the broth microdilution method and reported as minimal inhibitory concentration (MIC, ug/mL). AIMS OF THE STUDY: i) to investigate the traditional uses of arrayán (M. fragrans) in the South region of Ecuador; ii) to identify the main components of the essential oils isolated at different phenological stages; iiì) to test the antimicrobial activity of the essential oils against bacteria and yeasts causing human ailments and a yeast causing food spoilage. RESULTS: 37, 46 and 38 compounds, representing 96.5%, 96.2%, and 95.6% of the three essential oils (Fo, Fl and Fr), respectively, have been identified. Oxygenated monoterpenes (OM) were the major components with percentages of 63.1 (Fo), 49.4 (Fl), and 61.9% (Fr), respectively. The main constituents of the essential oils were the monoterpene aldehydes geranial (1) and neral (2), the content of which varied, depending on the phenological development stage of the plant, spanning from 31.1% and 23.6% (Fo), to 23.6% and 17.8% (Fl), and 29.7% and 24.3% (Fr), respectively. In vitro antimicrobial tests showed that the essential oils from M. fragrans exhibited good activity against the Gram-negative bacteria, K. pneumoniae, and against the yeasts, C. albicans and S. cerevisiae. CONCLUSIONS: The oil is characterized by a high concentration of the monoterpene aldehydes geranial and neral (citral), that make the aroma of colada morada prepared in southern Ecuador quite different from the beverage made in other regions of the country, where other types of myrtles (Myrtaceae spp.) are used. Moreover, the oil may become a new rich source of the important industrial chemical citral. The pleasant aromatic properties and the good in vitro antimicrobial activity of arrayán oil suggest a plausible scientific explanation for the traditional uses of the plant not only as a natural aromatizer of a traditional beverage but also as a natural anti-infective and anti-yeast agent.

Phytochemical Study of the Ecuadorian Species Lepechinia mutica (Benth.) Epling and High Antifungal Activity of Carnosol against Pyricularia oryzae.

The plant Lepechinia mutica (Benth.) Epling (family Lamiaceae) is endemic to Ecuador. In the present study, we report some major non-volatile secondary metabolites from the leaves and the chemistry of the essential oil distilled from the flowers. The main identified compounds were carnosol, viridiflorol, ursolic acid, oleanolic acid, chrysothol, and 5-hydroxy-4′,7-dimethoxy flavone. Their structures were determined by X-ray diffraction and NMR and MS techniques. The essential oil showed a chemical composition similar to that distilled from the leaves, but with some qualitative and quantitative differences regarding several minor compounds. The main constituents (>4%) were: δ-3-carene (24.23%), eudesm-7(11)-en-4-ol (13.02%), thujopsan-2-α-ol (11.90%), β-pinene (7.96%), valerianol (5.19%), and co-eluting limonene and β-phellandrene (4.47%). The volatile fraction was also submitted to enantioselective analysis on a β-cyclodextrin column, obtaining the separation and identification of the enantiomers for α-thujene, β-pinene, sabinene, α-phellandrene, limonene and β-phellandrene. Furthermore, the anti-fungal activity of non-volatile secondary metabolites was tested in vitro, with carnosol resulting in being very active against the “blast disease” caused by the fungus Pyricularia oryzae.

Phytochemical Investigation of Male and Female Hedyosmum scabrum (Ruiz & Pav.) Solms Leaves from Ecuador.

This article reports the chemical composition of the essential oils obtained by hydrodistillation of male and female H. scabrum fresh leaves. The essential oils, HSMO and HSFO, respectively, were analyzed by GC/MS and GC-FID. A total of 93 components were detected, accounting for 94.8% and 95.3% of HSMO and HSFO, respectively. The prevalent constituents of HSMO were pinocarvone (13.1%), d-germacren-4-ol (12.6%), 1,8-cineole (10.8%), α-pinene (6.4%), and ß-pinene (4.8%), whereas the major components of HSFO were 1,8-cineole (20.5%), linalool (16.5%), α-pinene (15.0%), ß-pinene (6.4%), and sabinene (6.3%). The different enantiomeric distribution of ß-pinene, sabinene, limonene, linalool in the two oils, was determined. The non-volatile esters of p-coumaric and ferulic acids with borneol (1 and 4), cis-chrysanthenol (2 and 5), and cis-pinocarveol (3 and 6) were identified in the leaves after basic hydrolysis and analysis of the NMR spectra of the free acids, and GC/MS spectra of the monoterpene alcohols, respectively. Compounds 2, 3, 5, and 6 have been found in nature for the first time. These findings demonstrated that, from a chemical point of view, male and female individuals of H. scabrum collected in Ecuador seem quite differentiated between each other and from samples of the same species growing in Bolivia and in Peru.

Chemical Composition, Enantiomeric Analysis, AEDA Sensorial Evaluation and Antifungal Activity of the Essential Oil from the Ecuadorian Plant Lepechinia mutica Benth (Lamiaceae).

This study describes the GC-FID, GC/MS, GC-O, and enantioselective GC analysis of the essential oil hydrodistilled from leaves of Lepechinica mutica (Lamiaceae), collected in Ecuador. GC-FID and GC/MS analyses allowed the characterization and quantification of 79 components, representing 97.3% of the total sample. Sesquiterpene hydrocarbons (38.50%) and monoterpene hydrocarbons (30.59%) were found to be the most abundant volatiles, while oxygenated sesquiterpenes (16.20%) and oxygenated monoterpenes (2.10%) were the minor components. In order to better characterize the oil aroma, the most important odorants, from the sensorial point of view, were identified by Aroma Extract Dilution Analysis (AEDA) GC-O. They were α-Pinene, ß-Phellandrene, and Dauca-5,8-diene, exhibiting the characteristic woody, herbaceus, and earthy odors, respectively. Enantioselective GC analysis of L. mutica essential oil revealed the presence of twelve couples and two enantiomerically pure chiral monoterpenoids. Their enantiomeric excesses were from a few percent units to 100%. Moreover, the essential oil exhibited moderate in vitro activity against five fungal strains, being especially effective against M. canis, which is a severe zoophilic dermatophyte causal agent of pet and human infections.

Phytochemical and ethnomedicinal study of Huperzia species used in the traditional medicine of Saraguros in Southern Ecuador; AChE and MAO inhibitory activity.

ETHNOBOTANICAL AND ETHNOMEDICINAL RELEVANCE: This study concerns seven Huperzia species (Lycopodiaceae), namely H. brevifolia, H. columnaris, H. compacta, H. crassa, H. espinosana, H. tetragona, H. weberbaueri, which are considered sacred plants by the Saraguro community, living in the Southern Andes of Ecuador; these plants are widely used in traditional medicine and ritual ceremonies. MATERIAL AND METHODS: The plants were selected on the basis of written interviews with 10 visionary healers (yachak) (2 women, 8 men), indicated as the most credible by the Saraguro Healers Council. The Informant Consensus Factor (Fic) was determined. The first phytochemical study of the plants was performed by standard procedures, while the AChE and MAO-A inhibition by fractions enriched in high MW alkaloids, was measured in vitro. AIMS OF THE STUDY: i) to investigate the uses of some Huperzia plants in healing and magical-religious practices of Saraguros; ii) to identify the main components of plant hydromethanolic extracts; iiì) to test the effects of alkaloidal fractions on the activity of two enzymes linked to mental health. RESULTS: All the interviewed Saraguro yachak showed a high consensus about the uses of the seven Huperzia plants as purgatives and against supernatural diseases, such as the "espanto" (startle). In admixtures with other plants, some species also induce a state of trance or hallucinations in participants in magical-religious rituals. GC-MS of the volatile alkaloid fractions allowed the identification of some lycodine-type and lycopodine-type alkaloids (1-5) in H. compacta, H. columnaris, and H. tetragona. The flavones selgin) (6) and tricin (7) were isolated from H. brevifolia and H. espinosana. Tricin (7) was also detected in the other five species. The rare serratene triterpenes serratenediol (8) serratenediol-3-O-acetate (9), 21-episerratenediol (10), and 21-episerratenediol-3-O-acetate (11) were isolated from H. crassa. In addition, the presence of an unprecedented group of high molecular weight alkaloids has been determined. Alkaloid fractions of H. brevifolia, H. compacta, H. espinosana, and H. tetragona significantly inhibited AChE and MAO-A activities in vitro. CONCLUSIONS: The first phytochemical and ethnopharmacological study of seven Huperzia plants, widely used by Saraguro healers, led to the identification of several alkaloids and triterpenoids with different remarkable biological activities. In addition, alkaloid fractions exhibited a significant AChE and MAO-A inhibitory activity. These results may support the use of these plants in brews prepared for inducing psychoactive effects in participants in magical-religious ceremonies. This study confirms the rich traditional medical knowledge of Saraguro healers which must be documented and preserved for future generations.