Combined Action of Piperitenone Epoxide and Antibiotics Against Clinical Isolates of Staphylococcus aureus and Escherichia coli.

The aim of this study was to determine the antimicrobial efficiency of piperitenone epoxide (PEO) - a principal component of various aromatic plants' essential oil - in combination with various antibiotics against 28 strains of Staphylococcus aureus and 10 strains of Escherichia coli isolated from clinical samples. Mentha spicata's essential oil, initially collected by hydrodistillation, was then subjected to flush column chromatography affording PEO of high purity. Minimum inhibitory concentrations of PEO alone and in combination with various concentrations of antibiotics were assessed using the microdilution method. The combined action was estimated calculating the fractional inhibitory concentration (FIC) index from checkerboard assays. Our results showed that the average minimum inhibitory concentration (mg/l) of PEO alone against E. coli was 512 ± 364.7 µg/ml, which was significantly higher than 172.8 ± 180.7 µg/ml observed for S. aureus. From checkerboard assays, FIC values below the 0.5 index, indicating synergy, were observed for 59% of the drugs tested. Twelve percent of FIC index values were between 0.5 and 1, indicating additive effects, while 21% were indifferent. According to our results, PEO could be a promising antimicrobial compound when combined with specific antibiotics and deserves further study.

Biocidal Compounds from Mentha sp. Essential Oils and Their Structure-Activity Relationships.

Essential oils from Greek Mentha species showed different chemical compositions for two populations of M. pulegium, characterized by piperitone and pulegone. Mentha spicata essential oil was characterized by endocyclic piperitenone epoxide, piperitone epoxide, and carvone. The bioactivities of these essential oils and their components have been tested against insect pests (Leptinotarsa decemlineata, Spodoptera littoralis and Myzus persicae), root-knot nematodes (Meloydogine javanica) and plants (Lactuca sativa, Lolium perenne, Solanum lycopersicum). The structure-activity relationships of these compounds have been studied including semi-synthetic endocyclic trans-carvone epoxide, exocyclic carvone epoxide, a new exocyclic piperitenone epoxide and trans-pulegone epoxide. Leptinotarsa decemlineata feeding was affected by piperitenone and piperitone epoxide. Spodoptera littoralis was affected by piperitone epoxide and pulegone. The strongest nematicidal agent was piperitenone epoxide, followed by piperitone epoxide, piperitenone and carvone. Germination of S. lycopersicum and L. perenne was significantly affected by piperitenone epoxide. This compound and carvone epoxide inhibited L. perenne root and leaf growth. Piperitenone epoxide also inhibited the root growth of S. lycopersicum. The presence of a C(1) epoxide resulted in strong antifeedant, nematicidal and phytotoxic compounds regardless of the C(4) substituent. New natural crop protectants could be developed through appropriate structural modifications in the p-menthane skeleton.

Bioefficacy of acyclic monoterpenes and their saturated derivatives against the West Nile vector Culex pipiens.

Twenty acyclic monoterpenes with different functional groups (acetoxy, hydroxyl, carbonyl and carboxyl) bearing a variable number of carbon double bonds were assayed as repellent and larvicidal agents against the West Nile vector Culex pipiens. Seven of them were derivatives that were synthesized through either hydrogenation or oxidation procedures. All repellent compounds were tested at the dose of 1mgcm(-2) and only neral and geranial were also tested at a 4-fold lower dose (0.25mgcm(-2)). Repellency results revealed that geranial, neral, nerol, citronellol, geranyl acetate and three more derivatives dihydrolinalool (3), dihydrocitronellol (5) and dihydrocitronellyl acetate (6) resulted in no landings. Based on the LC50 values the derivative dihydrocitronellyl acetate (6) was the most active of all, resulting in an LC50 value of 17.9mgL(-1). Linalyl acetate, citronellyl acetate, neryl acetate, geranyl acetate, dihydrocitronellol (5), dihydrocitronellal (7), citronellol, dihydrolinalyl acetate (2), citronellic acid and tetrahydrolinalyl acetate (1) were also toxic with LC50 values ranging from 23 to 45mgL(-1). Factors modulating toxicity have been identified, thus providing information on structural requirements for the selected acyclic monoterpenes. The acetoxy group enhanced toxicity, without being significantly affected by the unsaturation degree. Within esters, reduction of the vinyl group appears to decrease potency. Presence of a hydroxyl or carbonyl group resulted in increased activity but only in correlation to saturation degree. Branched alcohols proved ineffective compared to the corresponding linear isomers. Finally, as it concerns acids, data do not allow generalizations or correlations to be made.

Bioactivity of Dianthus caryophyllus, Lepidium sativum, Pimpinella anisum, and Illicium verum essential oils and their major components against the West Nile vector Culex pipiens.

Mosquitoes constitute a severe health problem in many areas all over the world. There are many regions of the tropics and subtropics where mosquitoes are one of the main reasons for inhibiting the economic upgrade. Except nuisance, their medical importance is another matter of attention since mosquitoes are vectors for a wide variety of vector-borne diseases. Due to disadvantages of currently used chemical control methods, it is unavoidable to search for eco-friendly new molecules. We report herein the evaluation of the larvicidal effect exhibited by essential oils of Dianthus caryophyllus, Lepidium sativum, Pimpinella anisum, and Illicium verum against late third to early fourth instar mosquito larvae of Culex pipiens. Furthermore, phytochemical analysis of plant samples revealed their major compounds to be ß-caryophyllene, eugenol, eucalyptol, α-terpinyl acetate, and (E)-anethole which were also tested for their potential larvicidal activity. For D. caryophyllus and L. sativum, this was the first report on the chemical composition of their essential oils. The essential oils of I. verum and P. anisum demonstrated high larvicidal activity with a LC(50) <18 mg L(-1). The other two essential oils of D. caryophyllus and L. sativum revealed moderate larvicidal activity, displaying a LC(50) value above 50 mg L(-1). Among the pure components, the most toxic were eugenol, (E)-anethole, and α-terpinyl acetate, with LC(50) values 18.28, 16.56, and 23.03 mg L(-1), respectively. Eucalyptol (1,8 cineole) and ß-caryophyllene were inactive at concentrations even as high as 100 mg L(-1), showing the least significant activity against mosquito larvae. Results allow some rationalization on the relative importance of the major compounds regarding the larvicidal activity of selected essential oils and their potential use as vector control agents.

Classification of Greek Mentha pulegium L. (Pennyroyal) samples, according to geographical location by Fourier transform infrared spectroscopy.

INTRODUCTION: Mentha pulegium L. (pennyroyal) is one of the four most commercially important Mentha species, even it is not a cultivated plant. It can be abundantly located in the Iberian Peninsula and North African countries. In Greece it grows in the wild and it is scattered all over the country. Pennyroyal is best known for its essential oil, with Spain and Morocco being the largest producers in the world. Mid-infrared spectroscopy has been applied to determine the origin of various samples. OBJECTIVES: In this work Fourier transform infrared spectroscopy (FT-IR) combined with canonical discriminant analysis has been applied to distinguish 70 Greek pennyroyal samples according to their collection areas. MATERIAL AND METHODS: Pennyroyal nonpolar organic extracts were prepared using ultrasound-assisted solvent extraction. The spectra of the extracts were recorded in the range of 4000-400 cm(-1) and the best discrimination was achieved in the spectral region 1720-1650 cm(-1) . RESULTS: Spectral features for the discrimination of pennyroyal samples among the different collection areas occur primarily in the carbonyl region and are correlated with the main volatile constituents of the extracts (menthone, isomenthone, pulegone, piperitone). All areas were easily differentiated by canonical discriminant analysis. The percentages of correct classification and validation were 94.3 and 90.0%, respectively. CONCLUSION: The combination of FT-IR spectroscopy and multivariate analysis provides a rapid and ambient method to discriminate pennyroyal samples in terms of geographical origin.

Quantitative determination of pulegone in pennyroyal oil by FT-IR spectroscopy.

Pulegone constitutes a monoterpene occurring in Mentha species and primarily in Mentha pulegium L. (pennyroyal). A major source of human exposure to pulegone is the use of pennyroyal essential oil in flavorings, confectionery and cosmetics. The rapid quantification of pulegone in hydrodistilled pennyroyal oils (which were also "spiked" to increase the validation range) by Fourier transform infrared spectroscopy (FT-IR) combined with partial least-squares (PLS) regression was evaluated, using the spectral region 1650-1260 cm(-1). Gas chromatography was applied as the reference method for pennyroyal oil samples, which ranged in pulegone content from 157 to 860 mg/mL. The two methods were subjected to statistical tests and proved equivalent in terms of accuracy and reproducibility (99% confidence level). The use of FT-IR spectroscopy could offer a viable alternative to the standard analysis procedures presently applied for quantification of valuable plant substances and could also provide the processing industry with a simple and high-throughput technique for the fast quality check of incoming raw materials such as pennyroyal oils.

Coactivity of sulfide ingredients: a new perspective of the larvicidal activity of garlic essential oil against mosquitoes.

BACKGROUND: The activity of several garlic oil ingredients against Culex pipiens L. larvae was studied. In addition to diallyl sulfide (DS) and diallyl disulfide (DDS), the garlic oils used included one essential oil (EO) and two semi-synthetic garlic essential oils (SSGEO1 and SSGEO2), which resulted from the enrichment of EO with DS and DDS standards respectively. The oils were compared with respect to their toxicity. RESULTS: The chemical composition of the tested oils was evaluated by means of gas chromatography-mass spectrometry. Experimental data from the tested samples revealed high toxicity. In detail, DDS (6.09 mg L(-1)) was the most active, followed by SSGEO2 (7.05 mg L(-1)) and EO (8.01 mg L(-1)), while SSGEO1 and DS were relatively inactive. CONCLUSION: The addition of DDS to EO did not change the toxic effect of the essential oil, whereas the presence of DS in excess produced an enriched essential oil with low toxicity. Furthermore, the antagonistic effect of DS and DDS against the other components of EO was shown by the application of two equations obtained from the literature. Their larvicidal performances were correlated, for the first time, with their detailed chemical composition. Data analysis strongly indicated the toxicity of the other EO sulfur ingredients.

Comparison of distillation and ultrasound-assisted extraction methods for the isolation of sensitive aroma compounds from garlic (Allium sativum).

A comparative study of traditional simultaneous distillation extraction (SDE), microwave assisted hydrodistillation extraction (MWHD) and ultrasound-assisted extraction (USE) is presented, for the extraction of essential oils from fresh garlic (Allium sativum) cloves. Each method is evaluated in terms of qualitative and quantitative composition of the isolated essential oil. The highly reactive sulfur molecules of the garlic volatile fraction show variable response to the different isolation methods. The application of ultrasound for the extraction of the essential oil is considered to cause a lesser damage of thermal-sensitive molecules, thus, providing a better approach of the compounds primarily responsible for the characteristic odor and taste of freshly chopped garlic. All heat-involving isolation procedures have been shown to differentiate the volatile-fraction profile as analyzed by GC-MS. Especially when grouping the compounds into cyclic and acyclic, the percentage concentrations drop from 77.4% to 8.7% for the acyclic while that of the cyclic compounds increase from 4.7% to 70.8%. The observed fact may be attributed to the effect of the heat applied, which changes from harsh thermal treatment (SDE) to short time thermal (MWHD) and room-temperature isolation (USE). The use of USE proves to be crucial in order to provide reliable insight into garlic's chemistry.

Rapid method for simultaneous quantitative determination of four major essential oil components from oregano (Oreganumsp.) and thyme (Thymus sp.) using FT-Raman spectroscopy.

A simple and rapid method for the quantitative determination of four major components found in oregano and thyme essential oils is presented. The method correlates the Raman peak intensity in the spectral region from 1800 to 600 cm(-1) and the concentration percentage of each particular constituent in the sample. To achieve accurate quantification results and avoid the risk of overlapping peaks of unknown Raman-active substances in natural essential oils, the peaks must be analyzed. For this purpose, PEAKSOLVE software (Ver. 1.0.5) was used. Unknown samples were measured with the FT-Raman method, and the results were compared to those of the gas chromatographic (GC) analysis. The comparison was made at a confidence level of 99%, and the two methods scored equally in terms of repeatability and accuracy even at the edge of the method specifications. The new method can provide accurate results in very short times once the setup is complete and could be utilized in areas where vast amounts of samples must be analyzed.

FT-Raman spectroscopic simultaneous determination of fructose and glucose in honey.

A new method for mass percentage determination of fructose and glucose based on FT-Raman spectroscopy is evaluated with a standard HPLC-based method. FT-Raman spectra manipulation is done via the spectrometer software, and a PLS (partial least squares) method is developed with the TQ Analyst software (Ver 1. 1a). The simultaneous quantitative determination uses an input range from 1700 to 700 cm(-1) without correction or baseline factors. The standards used in the PLS method are honey samples previously analyzed by HPLC to obtain their mass percentage concentrations in fructose and glucose. The returned results are statistically tested with those of the HPLC method. Both methods appear to score equally in terms of reproducibility. The honey content of the two sugars in total was found up to 40-74%. The honey samples content in fructose and glucose was determined by HPLC (24.1-42.9% and 16.2-33.1%, respectively) and FT-Raman (24.0-40.8% and 21.1-32.2%, respectively).