Essential oil composition and biological activities determination of Mosla dianthera (Buch.-Ham. ex Roxb.) Maxim. and its major isolated component, carvone

Abstract This study was aimed to explore the chemical composition and biological activities of essential oil from aerial part of Mosla dianthera along with its major isolated compound, carvone. The hydro-distilled essential oil was analysed by GC-MS and biological activities were investigated in terms of antioxidant, anti-inflammatory, herbicidal, antibacterial, anti-fungal and anti-feedant properties. GC-MS analysis led to the identification of forty-nine components contributing 96.2% of essential oil with carvone (41.9%) as the most abundant constituent. The oil and carvone showed good to moderate antioxidant potentials determined by radical scavenging, reducing power and metal chelating activities. Carvone showed good anti-inflammatory activity (78.0%) compared to essential oil (74.2%). Both essential oil and carvone exhibited excellent herbicidal activity against Raphanus raphanistrum subsp. sativus seeds. The essential oil and carvone showed significant anti-bacterial efficacy against Bacillus cereus and Escherichia coli. It was observed that essential oil showed strong antifungal property than carvone against Alternaria alternata and Curvularia lunata. Both the samples exhibited anti-feedant activity in a dose dependent manner against third instar larvae of Spilosoma obliqua. Results obtained revealed the possible applications of essential oil and carvone as a bioactive source of natural antioxidants, excellent herbicide and an effective substance for antifungal and antifeedant activities.

Study on peel components and juice quality of three mandarin hybrids (Citrus reticulata) on sour orange rootstock cultivated in Ramsar, Iran / Estudio sobre los componentes de la cáscara y la calidad del jugo de tres híbridos de mandarina (Citrus reticulata) en patrones de naranja agria cultivados en Ramsar, Irán

The peel components and juice quality of three mandarin hybrids were investigated in this study. Peel components were extracted using the cold-press method and analyzed using GC-FID and GC-MS. Total soluble solids, total acid, pH value, ascorbic acid as well as density were determined in juice obtained from mandarin hybrids. Twenty-six, thirty-five and nineteen peel components were identified in Fortune, Robinson and Osceola respectively including: aldehydes, alcohols, esters, monoterpenes, sesquiterpenes and other components. The major components were limonene, γ-terpinene, (E)-ß-ocimene, ß-myrcene, sabinene, linalool and α-Pinene. Among the three scions examined, Fortune showed the highest content of aldehydes and Robinson showed the highest content of TSS. This study shows that scion has a profound influence on aldehyd and TSS that are important to quality improvement.

Los componentes de la cáscara y la calidad del jugo de tres híbridos de mandarina fueron investigados en este estudio. Los componentes de la cáscara fueron extraídos por el método de prensado en frío y se analizaron mediante GC-FID y GC-MS. el total de sólidos solubles, acidez total, pH, ácido ascórbico, así como la densidad se determinaron en el jugo obtenido a partir de híbridos de mandarina. Veintiséis, treinta y cinco y diecinueve componentes de la cáscara de Fortuna, Robinson y Osceola respectivamente fueron identificados, incluyendo: aldehídos, alcoholes, ésteres, monoterpenos, sesquiterpenos y otros componentes. Los componentes principales eran limoneno, γ-terpineno, (E) -ß-ocimeno, ß-mirceno, sabineno, linalool y α-pineno. Entre los tres vástagos examinados, Fortune mostró el mayor contenido de aldehídos y Robinson mostró el mayor contenido de SST. Este estudio muestra que el vástago tiene una profunda influencia en aldehído y SST que son importantes para mejorar la calidad.

Evaluation of antimicrobial, cytotoxic and chemopreventive activities of carvone and its derivatives

ABSTRACT Considering the reported activity of carvone in the literature, this study aimed to evaluate the antimicrobial, cytotoxic and chemopreventive activities of (+)- and (-)-carvone, (+)- and (-)- hydroxydihydrocarvone and α,ß-epoxycarvone. (+)-Hydroxydihydrocarvone (HC+), (-)-hydroxydihydrocarvone (HC-) and α,ß-epoxycarvone (EP) were obtained by synthesis using (+)-carvone (C+) or (-)-carvone (C-) as precursors. The antifungal activity (MIC and MFC) were evaluated against Candida parapsilosis, C. tropicalis, C. krusei and C. albicans and the antibacterial activity (MIC and MBC) against Escherichia coli and Staphylococcus aureus. The cytotoxicity assays were performed with human cancer cell lines HepG-2 and SiHa and the normal strain MRC-5 through sulphorrodamine B assay. Chemoprevention was evaluated through quinone reductase assay. Our results showed no cytotoxicity on tumor and normal cell lines and no induction of the quinone reductase enzyme. C- and HC- presented activity against E. coli. All compounds presented weak antifungal activity against C. tropicalis and C. parapsilosis. EP and C+ showed moderate activity against C. krusei. Results suggest the potential use of carvones and its derivatives as antifungal agents against Candida yeasts. The absence of cytotoxicity in cell lines indicates safety in the use of these compounds

Antibacterial activity of essential oils of aloysia polystachya and lippia turbinata (Verbenaceae) / Actividad antibacteriana de los aceites esenciales de aloysia polystachya y lippia turbinata (Verbenaceae)

Aloysia polystachya and Lippia turbinata are medicinal and aromatic plants. Nevertheless, there are few reports in literature concerning the biological properties of species that grow in northeastern Argentina. The antibacterial activity and the chemical composition of both essential oils were evaluated in this work. The extraction was performed by steam distillation and their volatile compounds were determined by gas chromatography/mass spectrometry. The antibacterial activity was evaluated by disc diffusion and broth microdilution assay. The main compounds were carvone (78.9 percent) and limonene (14.2 percent) in A. polystachya and carvone (80.77 percent), limonene (8.73 percent), beta- caryophyllene (2.13 percent) and 1,8-cineole (1.70 percent) in L. turbinata. Both essential oils were bactericide against Escherichia coli ATCC 35218 and clinical isolates of Enterobacter cloacae and Klebsiella pneumoniae. Essential oil of A. polystachya was also bactericide against Staphylococcus aureus ATCC 29212, S. aureus ATCC 25923 and clinical strain of S. aureus methicillin susceptible.

Aloysia polystachya y Lippia turbinata son plantas medicinales y aromáticas. Hay pocos informes en la literatura sobre las propiedades biológicas de especies que crecen en el nordeste de Argentina. La actividad antibacteriana y la composición química de ambas especies se evaluaron en este trabajo. La extracción se realizó por destilación con vapor y sus compuestos se determinaron por cromatografía gaseosa/espectrometría de masa. La actividad antibacteriana fue evaluada por difusión en discos y microdilución en caldo. Los principales compuestos fueron carvona (78.9 por ciento) y limoneno (14.2 por ciento) en A. polystachya y carvona (80.77 por ciento), limoneno (8.73 por ciento), beta-cariofileno (2.13 por ciento) y 1,8-cineol (1.70 por ciento) en L. turbinata. Ambos aceites esenciales fueron activos contra Escherichia coli ATCC 35218 y aislamientos clínicos de Enterobacter cloacae y Klebsiella pneumoniae. El aceite esencial de A. polystachya fue bactericida contra Staphylococcus aureus ATCC 29212, S. aureus ATCC 25923 y aislamientos clínicos de S. aureus sensible a meticilina.

Composition, anti-quorum sensing and antimicrobial activity of essential oils from Lippia alba

Many Gram-negative pathogens have the ability to produce N-acylhomoserine lactones (AHLs) as signal molecules for quorum sensing (QS). This cell-cell communication system allows them to coordinate gene expression and regulate virulence. Strategies to inhibit QS are promising for the control of infectious diseases or antibiotic resistant bacterial pathogens. The aim of the present study was to evaluate the anti-quorum sensing (anti-QS) and antibacterial potential of five essential oils isolated from Lippia alba on the Tn-5 mutant of Chromobacterium violaceum CV026, and on the growth of the gram-positive bacteria S. aureus ATCC 25923. The anti-QS activity was detected through the inhibition of the QS-controlled violacein pigment production by the sensor bacteria. Results showed that two essential oils from L. alba, one containing the greatest geranial:neral and the other the highest limonene:carvone concentrations, were the most effective QS inhibitors. Both oils also had small effects on cell growth. Moreover, the geranial/neral chemotype oil also produced the maximum zone of growth inhibition against S. aureus ATCC 25923. These data suggest essential oils from L. alba have promising properties as QS modulators, and present antibacterial activity on S. aureus.

Lippia alba morphotypes cidreira and melissa exhibit significant differences in leaf characteristics and essential oil profile

Lippia alba (Mill.) N.E. Br. ex Britton & P. Wilson, Verbenaceae, is widely used in traditional Brazilian medicine for the treatment of abdominal distress. The species exhibits considerable chemical and morphological diversity, and various chemotypes have been characterized. A comparative study of L. alba, has been carried out of the morphoanatomical characteristics of the leaves and the profiles of the essential oils of the morphotypes cidreira and melissa grown in the Medicinal Plant Garden of the Universidade Estadual de Santa Cruz, Ilhéus, Bahia, Brazil. The mean plant height of cidreira was 1.80 m and the stems and branches were fairly erect, while melissa plants were smaller (1.60 m) and presented prostrate stems and branches. Although the leaf of the morphotypes look were similar, the mean values of length, width and area of the leaves of cidreira (respectively, 7.42 cm, 3.32 cm and 17.31 cm²) differed significantly from those of melissa (4.68 cm, 2.35 cm and 7.32 cm2). The morphotypes presented amphistomatic leaves with uniseriate epidermis on both surfaces. The mesophyll was dorsiventral, but in cidreira the palisade parenchyma was biseriate while in melissa it was uniseriate. Simple tector and capitate glandular trichomes were present on the adaxial and abaxial surfaces of the leaf blades of both morphotypes. Six distinct types of glandular trichomes could be distinguished: types I and II were present in both morphotypes, while type III was detected only in cidreira, and types IV to VI were present only in melissa. The two morphotypes also differed with respect to the composition of the essential oil, cidreira produced oil composed mainly of citral, while the oil from melissa was rich in citral, limonene and carvone.

Anxiolytic effects of repeated treatment with an essential oil from Lippia alba and (R)-(-)-carvone in the elevated T-maze

Lippia alba (Mill.) N.E. Brown (Verbenaceae) is widely used in different regions of Central and South America as a tranquilizer. The plant’s anxiolytic properties, however, merit investigation. The present study evaluated the effects of repeated daily (14 days) intraperitoneal (ip) treatment with an essential oil (EO) from a chemotype of L. alba (LA, chemotype II, 12.5 and 25 mg/kg; N = 6-8) and (R)-(-)-carvone (25 mg/kg; N = 8-12), the main constituent of this chemotype, on male Wistar rats (weighing 250 g at the beginning of the experiments) submitted to the elevated T-maze (ETM). The ETM allows the measurement of two defensive responses: inhibitory avoidance and one-way escape. In terms of psychopathology, these responses have been related to generalized anxiety and panic disorder, respectively. Treatment with the EO impaired ETM avoidance latencies, without altering escape, in a way similar to the reference drug diazepam (P < 0.05) (avoidance 2: control = 84.6 ± 35.2; EO 12.5 mg/kg = 11.8 ± 3.8; EO 25 mg/kg = 14.6 ± 2.7; diazepam = 7 ± 2.1). (R)-(-)-carvone also significantly altered this same response (P < 0.05; avoidance 1: control = 91.9 ± 31.5; carvone = 11.6 ± 1.8; diazepam = 8.1 ± 3.3). These results were not due to motor changes since no significant effects were detected in an open field. These observations suggest that LA exerts anxiolytic-like effects on a specific subset of defensive behaviors that have been implicated in generalized anxiety disorder, and suggest that carvone is one of the constituents of LA responsible for its action as a tranquilizer.

Antiulcer effect of epoxy-carvone

Epoxy-carvone is a monoterpene present in essential oils of various plants. It is a derivative of carvone, which has an epoxy group instead of the α- and β-unsaturated ketone group present in carvone. As recent studies have shown that several alcohol terpenes and compounds containing α, β-unsaturated ketone groups present antiulcer effect, the main of the present study was to evaluate the antiulcer effect of epoxy-carvone. The models of ulcers induced by ethanol and indomethacin were used in this study. Epoxy-carvone at the dose of 1 mg/kg did not present antiulcer effect against ulcer induced by ethanol, but at the doses of 10, 30 and 50 mg/kg it presented gastroprotective effect in both ulcer models. Epoxy-carvone also did not affect the gastric secretion in the pylorus ligation test. Moreover, pretreatment with indomethacin or L-nitroarginine methyl ester did not reverse the gastroprotection produced by this monoterpene. This study showed that epoxy-carvone presents antiulcer effect and suggests that this effect does not involve either antisecretory activity or increase of the nitric oxide and prostaglandin synthesis.

Antispasmodic effects of Aloysia polystachya and A. gratissima tinctures and extracts are due to non-competitive inhibition of intestinal contractility induced by acethylcholine and calcium

The antispasmodic effects of acqueous extracts (AE) and tinctures (T) of Aloysia polystachya (Griseb.) Moldenke and Aloysia gratissima (Gillies & Hook.) Tronc., Verbenaceae, were studied on rat isolated ileum and duodenum. These plants are used for gastrointestinal disorders and as eupeptic in South America. Both AE non-competitively inhibited the dose-response curves (DRC) of ACh and the DRC of Ca2+ in high-[K+]o, as well as the T. The T of A. polystachya and A. gratissima respectively inhibited the ACh-DRC at the IC50 of 3.15±0.57 and 6.46±2.28 mg leaves/mL. The Ca2+- antagonist activity of both T occurred with IC50 respectively similar to those of the ACh-DRC, and was potentiated by the depolarization produced by 10 mM TEA, a blocker of K+- channels. The spasmolytic effect of T does not involve DA release and binding to D2, since it was not reduced by 10 µ M metoclopramide. Also, T induced dose-dependent relaxation on the tonic contracture produced by high -[K+]o and ACh. By TLC there were detected in the leaves the presence of carvone, and flavonoids such as quercetin and hesperidin. By HPLC there were not found vitexin nor isovitexin, identified in A. citriodora. The monoterpene (-)- carvone non-competitively inhibited the ACh-DRC (pD'2 of 4.0±0.1) and the DRC of Ca2+ (pD'2 of 3.86±0.19), suggesting that the Ca2+- influx blockade is the mechanism of its antispasmodic effect. Results suggest that the antispasmodic effect of A. polystachya and A. gratissima are mostly explained by the non-competitive blockade of Ca+2 influx. It could be associated to the presence of flavonoids, and in the tinctures to some spasmolytic components of the essential oil such as carvone.

Water soluble fractions of caraway (Carum carvi L. ) essential oil / Fracción de aceite esencial de alcaravea (Carum carvi L. ) soluble en agua

Natural essential oils are used extensively in fragrances, flavorants, and in the food and pharmaceutical industries. During hydrodistillation, a part of the essential oil becomes dissolved in the condensate and lost as this water is discarded. In this study, carvone and limonene content recovered from hydrodistillation waste water of caraway fruit were quantified using two methods for recovering dissolved aromatic molecules from condensate water: extraction through distillation and extraction by means of a solvent. This allows for the conservation of useful molecules which are typically discarded with the waste water produced during the distillation process. The objective of this study was to quantify the carvone and limonene content recoverable from waste water derived from the distillation of caraway essential oil. The well-known Clevenger method and a simpler, more practical technique employing cyclohexane as a solvent were employed to determine the recoverable content of aromatic molecules from the hydrosol. The chemical compositions of the respective recovered extracts were compared with those of the primary oils to analyze the efficacy of these methods. Recovered extract accounted for 10 to 40 percent of the total oil yield. The limonene and carvone molecules recovered using these methods were quantified through gas chromatography in order to characterize the composition of the secondary extract produced.

Los aceites esenciales naturales se utilizan ampliamente en las fragancias, saborizantes, y en la industria alimentaria y farmacéutica. Durante la hidrodestilación, una parte del aceite esencial se disuelve en el condensado y se pierde como agua de descarga. En este estudio, el contenido de carvona y limoneno recuperados del agua de desecho de la hidrodestilación de la fruta de alcaravea se cuantificaron utilizando dos métodos para recuperar las moléculas aromáticas disueltas en el agua condensada: extracción a través de la destilación y la extracción con un disolvente. Esto permite la conservación de las moléculas útiles que normalmente son desechadas con las aguas residuales producidas durante el proceso de destilación. El objetivo de este estudio fue cuantificar el contenido de carvona y limoneno recuperable de las aguas residuales procedente de la destilación del aceite esencial de alcaravea. El conocido método de Clevenger y una técnica sencilla y práctica que emplea ciclohexano como disolvente fueron utilizadas para determinar el contenido de moléculas aromáticas contenidas en el hidrosol. La composición química de los extractos recuperados fue comparada con los aceites primarios para analizar la eficacia de estos métodos. El extracto recuperado representa del 10 al 40 por ciento del contenido total de aceite esencial. Las moléculas de limoneno y carvona recuperadas mediante estos métodos se cuantificaron mediante cromatografía de gases con el fin de caracterizar la composición del extracto secundario.

Análise do óleo essencial de folhas de três quimiotipos de Lippia alba (Mill.) N. E. Br. (Verbenaceae) cultivados em condições semelhantes / Analysis of the essential oil from leaves of three Lippia alba (Mill.) N. E. Br. (Verbenaceae) chemotypes cultivated on the same conditions

A composição do óleo essencial de Lippia alba apresenta variação quantitativa e qualitativa, levando à separação em quimiotipos. O trabalho tem como objetivo analisar o óleo essencial de folhas de três quimiotipos de Lippia alba, provenientes de diferentes regiões do Brasil, cultivados em condições semelhantes, a fim de verificar se as diferenças na composição do óleo devem-se a fatores ambientais ou a variação genética infraespecífica e se a floração influencia o rendimento e a composição do óleo. Os quimiotipos produtores de citral, carvona e linalol, foram denominados Lippia alba 1, 2 e 3, respectivamente. Os óleos essenciais foram extraídos por hidrodestilação de folhas e analisados por cromatografia com fase gasosa e cromatografia com fase gasosa acoplada ao espectrômetro de massas. O melhor rendimento foi obtido das plantas no estágio vegetativo. A composição do óleo essencial manteve-se inalterada para os três quimiotipos após cultivo em condições semelhantes e também não variou qualitativamente durante o crescimento vegetativo e floração. Os dados obtidos reforçam a idéia que as diferenças na composição do óleo essencial dos quimiotipos refletem variações genotípicas entre as plantas e que a extração de óleo essencial de L. alba deve ser efetuada na fase de crescimento vegetativo, quando é maior o rendimento do óleo e a porcentagem dos componentes majoritários.

The composition of Lippia alba essential oil varies in a manner that different chemotypes are recognized. This work deals with the analysis of the essential oil from three chemotypes of L. alba from different regions of Brazil, cultivated in similar conditions, to verify if the differences in their chemical composition can be due to environmental conditions or may be due to infra-specific genetic variation. The influence of flowering was investigated regarding essential oil yield and composition. The chemotypes producing citral, carvone and linalol were denominated chemotypes 1, 2 and 3, respectively. The essential oils were obtained by hydrodistillation of the leaves, at flowering and vegetative growth stages. Essential oils were analyzed by GC and GC/MS. A better yield was obtained from plants at vegetative stage. The composition of the essential oils remained unchanged after cultivation in the same conditions, and did not vary qualitatively during flowering and vegetative periods. Relative percentages of the major compounds changed during flowering period. Data obtained allow us to reinforce the idea that differences in the essential oil composition of the three chemotypes are due to genotypic variations and that the essential oil extraction during the vegetative period enables a better yield and higher percentages of the major compounds.