Use and abuse of retention indices in gas chromatography.

The value of the concept of retention indices (RI) to the practice of gas chromatography (GC) is highlighted, where the RI of a compound is one component of the strategy to identify the compound. The widespread reliance on GC and then on mass spectrometry for 'identification', may result in inadequate confirmation of molecular identity. However, RI do provide a useful tentative indication of the possible molecule(s). Thus, the RI value is a useful first measure of the molecule identity, and shown here to be valuable provided limitations are recognised. An author has a responsibility to correctly calculate the index and then use the values for (tentative) identification. Tables of reference RI values are useful in this respect, but finding an 'exact match' RI value does not confirm the identity. Hence, it is necessary to understand how the RI value may be incorrectly used in this respect. The reviewer of written research is charged with ensuring the index values are applied in a rigorous manner. Selected case studies from our own work, support the care that must be exercised when reporting RI values. In terms of advanced GC operations, mention is made of multidimensional gas chromatography and comprehensive two-dimensional gas chromatography to acquire RI values on both the first and second columns in the two-column separation experiment.

Synergism in Two-Component Insecticides with Dillapiole against Fall Armyworm.

The fall armyworm (Spodoptera frugiperda), a polyphagous insect pest, is a major threat to food production, rapidly spreading through all the tropical areas in the world. Resistance has developed to the control protocols used so far (pyrethroids, organophosphorus, and genetically modified plants), and alternative strategies must be found. The bioactivity in essential oils is usually associated with the major constituents, but synergistic interactions among the constituents (even minor ones) can improve the levels of activity considerably. Herein, we tested the insecticidal activity of several constituents of the essential oil from Piper aduncum, an Amazonian Piperaceae, both separately and as binary mixtures, through their application on the dorsal side of the larva pronotum. Dillapiole proved to be, isolated, the most active compound in this oil (LD50 = 0.35 ppm). In binary mixtures, a strong synergistic effect was observed for the pairs of dillapiole with ß-caryophyllene (LD50 = 0.03 ppm), methyl eugenol (LD50 = 0.05 ppm), and α-humulene (LD50 = 0.05 ppm). In some cases, however, antagonism was recorded, as for dillapiole + ß-pinene (LD50 = 0.44 ppm). The use of binary mixtures of essential oil constituents as low-environmental-toxicity insecticides allows a fine tuning of the insecticidal activity, and the exploitation of synergy effects.

Microencapsulation of marjoram essential oil as a food additive using sodium alginate and whey protein isolate.

Encapsulation techniques are generally used to preserve the volatile compounds of essential oils. This study aimed to evaluate the influence of process variables on the microencapsulation of marjoram essential oil (MEO) (Origanum majorana L.) by ionic gelation. The effect of sodium alginate concentration (0.5-2 g/100 mL), emulsifier concentration (0.5-2 g/100 mL whey protein isolate (WPI)), and cationic bath concentration (0.05-0.3 mol/L CaCl2) on the emulsions and beads properties were investigated, according to a rotatable central composite design. MEO chemical composition and antimicrobial activity were assessed. Emulsions were characterized for droplet size and viscosity, while the particles were analyzed for encapsulation efficiency, size and circularity, and morphology. High concentrations of alginate and WPI intensified the porous structure of the beads, reducing droplet mean diameter and encapsulation efficiency. High alginate concentrations also increased emulsion viscosity, affecting positively beads' circularity. The intermediate concentration of sodium alginate (1.25 g/100 mL), WPI (1.25 g/100 mL), and CaCl2 (0.175 mol/L) were selected as the most appropriate conditions to produce beads with satisfactory circularity and high encapsulation efficiency.

Sustainable use of tropical fruits? Challenges and opportunities of applying the waste-to-value concept to international value chains.

Agriculture and food science literature on waste-to-value applications that allow upcycling of by-product ingredients is increasing. However, this stream of research rarely takes an international trade and sustainability systems perspective. This focused review defines the term of waste-to-value and the sustainable development goals connected to it, and points to the tensions and questions arising when international trade is involved. Further, it exemplifies the challenges and opportunities of waste-to-value in tropical fruit trade through five cases of tropical fruit from South America: Green coconut, açaí, maracujá, cambuci, and jabuticaba. We present a model of the international supply chain that indicates where the opportunities of waste-to-value applications in international tropical fruit trade are situated, and discuss which future research questions need to be addressed to tackle the challenges of waste-to-value in global tropical fruit chains. Establishing the waste-to-value approach in the export of yet-underused tropical fruits can amongst others improve local employment, preserve natural resources, allow favorable use of side-streams in local energy production, environmentally friendly packaging material for transport, and add health functionalities to the end-consumer products, but challenges have to be solved in order to ensure these environmental and social benefits materialize.

Heteromorphic stamens are differentially attractive in Swartzia (Fabaceae).

The division of labour hypothesis between stamens has explained the evolution of divergent functions between dimorphic stamens in the same flower. However, little is known about whether the distinct type of stamens differs in attractiveness to pollinators. Therefore, we investigate whether the two types of stamens commonly found in Swartzia have different visual and olfactory attractants. We performed observations of anthesis dynamics, registration and collection of floral visitors, measurements of reflectance of floral parts and chemical analysis of the volatile organic compounds of the floral parts of two species, S. flaemingii and S. simplex. Both species have two distinct sets of stamens: one with smaller and abundant stamens in the centre of the flower and the other with fewer but larger abaxial stamens. The sets differ in UV reflectance (only S. simplex) and exhibit a distinct chromatic contrast. Concerning olfactory attractiveness, aliphatic compounds make up most of the odour of the two species, both whole flowers and most of their floral organs. On the other hand, only S. simplex presented apocarotenoids (as ionones) and benzenoids. Furthermore, there are differences in the proportion of volatiles emitted by the stamen in both cases, as the high proportion of sesquiterpenes among the smaller stamens compared to the larger ones. In conclusion, the two types of stamens found in S. flaemingii and S. simplex show a distinct attractiveness. In addition, our data have demonstrated diverse ways of differential attractiveness both between distinct stamens set per flower and between the two species from the same pollen flowers genus.

Acaricidal and anthelmintic action of ethanolic extract and essential oil of Achyrocline satureioides.

Although Brazilian livestock is considered one of the most productive in the world, the losses caused by parasites still greatly limit these activities and its incorrect control through traditional synthetic chemicals has caused problems such as parasitic resistance. The present study evaluated the anthelmintic and acaricidal action of ethanolic extract (EE) and essential oil (OE) obtained from Achyrocline satureioides inflorescences against an endo and an ectoparasite of great importance in sheep and cattle. The action against Haemonchus contortus was evaluated by the Larval Development Test (LDT) and the Egg Hatching Test (EHT), while the acaricidal potential against Rhipicephalus (Boophilus) microplus was evaluated by the Adult Immersion Test (AIT) and Larval Package Test (LPT). Chemical analyzes were performed by liquid chromatography coupled to mass of spectrometry (LC-MS) and gas chromatography coupled to mass spectrometry (GC-MS) in order to evaluate the chemical composition of EE and EO, respectively. Luteolin (m/z 285), quercetin (m/z 301) and methyl quercetin (m/z 315) were the major compounds detected in the A. satureioides EE, while 41 compounds were identified in EO. Among them, the major compounds were caryophyllene oxide (15.4%), limonene (10.9%), trans-calamenene + Î´-cadinene (8.0%), (E)-caryophyllene (5.5%), spathulenol (5.1%) and α-pinene (4.5%). Against H. contortus, by EHT, the EE and EO showed LC50 of 83.14 mg mL-1 and 10.42 mg mL-1, respectively. In LDT, LC50 were 0.16 mg mL-1 and 0.42 mg mL-1, respectively by EE and EO. In the AIT assay for R. (B.) microplus tick LC50 obtained for EE and EO were 76.43 mg mL-1 and 114.35 mg mL-1, respectively. EE at 100 mg mL-1 completely inhibited egg laying, while the EO at 50 mg mL-1 completely inhibited larval hatching. No significant larvicidal effect was observed in LPT for both extracts. The EE and EO presented a high efficacy at low concentrations in the inhibition of egg hatching and larval development of H. contortus. Although both extracts did not show consistent larvicidal action, the results were significant in inhibiting the oviposition of R. (B.) microplus engorged females.

Immobilization of phosphonium-based ionic liquid stationary phases extends their operative range to routine applications in the flavor, fragrance and natural product fields.

Phosphonium-based ionic liquids (ILs) have proven to be successful stationary phases (SPs) for gas chromatography (GC) in several fields of application because of their unique selectivity and good chromatographic properties. This study focuses on the use of two ILs as GC SPs that are based on the phosphonium derivatives trihexyl(tetradecyl)phosphonium chloride ([P66614+] [Cl-]), and trihexyl(tetradecyl)phosphonium bis[(trifluoromethyl)sulfonyl]imide ([P66614+][NTf2-]), which have previously been shown to be complementary in terms of chromatographic selectivity and retention. Their application in routine analysis has been limited by their lower maximum allowable operating temperatures (MAOT) (200 °C for the [P66614+][Cl-] IL and 180 °C for [P66614+][NTf2-]), which restricts their use to samples that consist of analytes with relatively high volatility. A previous study carried out in the Authors' laboratory focused on extending the use of the [P66614+][Cl-] IL SP to the analysis of samples with analytes of medium-to-low volatility by optimizing column characteristics and operative conditions. This study addresses the immobilization of both the [P66614+][Cl-] and [P66614+][NTf2-] ILs to the inner wall of fused silica columns to increase their MAOT under soft and hard reaction conditions. The resulting MAOT depended on more or less drastic immobilization conditions, and reached 220 °C for soft immobilization (So-Im) and 240 °C for hard immobilization (Ha-Im) in the [P66614+][Cl-] IL columns, and 200 °C for So-Im and 220° for Ha-Im in columns coated with the [P66614+] [NTf2-] IL. The influence of immobilization on the separation power and performance of all the columns has been evaluated using i) the Grob test, ii) a model mixture of 41 compounds of different polarity, structure, and with different organic functional groups representative of the flavor and fragrance field, iii) a standard mixture of 37 fatty acid methyl esters, iv) the peppermint essential oil, v) two mixtures of sesquiterpenic alcohols (farnesols and santalols), and vi) a standard mixture of 16 pesticides. These test samples were also used to demonstrate the complementarity of the two phosphonium-based IL SPs in terms of selectivity and retention.

Amazonian medicinal smokes: Chemical analysis of Burseraceae pitch (breu) oleoresin smokes and insights into their use on headache.

ETHNOPHARMACOLOGICAL RELEVANCE: In Latin American Traditional Medicine, the use of Burseraceae oleoresins for headache relief is widespread. In the quilombola (maroon) communities of the municipality of Oriximiná, Pará State, Brazil, Burseraceae oleoresins are burned as incenses to treat headache; an effect attained by inhaling the smokes. AIM OF THE STUDY: This study was designed to investigate the scientific rationale behind the use of Burseraceae pitch oleoresin smokes on headache by identifying its chemical constituents and discussing whether they could be beneficial for headache treatment. MATERIALS AND METHODS: Two samples of pitch oleoresins were purchased from Amazonian public markets; one of them marketed as "breu preto" (black pitch), and the other as "breu branco" (white pitch). The smoke headspaces produced upon burning these oleoresins were collected and analysed by GC-MS. For comparative purposes, the triterpenoid fingerprints of the oleoresins' dichloromethane extracts and profiles of the essential oils were also obtained by GC-MS - and GC-FID, in the case of the essential oils. RESULTS: A total of 32 compounds were detected in black and white pitch oleoresin smokes. All smoke headspaces of the pitch oleoresins shared the presence of volatile terpenoids (e.g., α-terpineol) and triterpenoids (e.g., α-amyrin, ß-amyrin, α-amyrone, and ß-amyrone). These compounds were also present in the crude resins and could potentially be responsible for anti-inflammatory, antinociceptive and analgesic effects on headache. CONCLUSIONS: The pharmacological data on the terpenoids detected in the Burseraceae pitch smokes strongly support their traditional use for headache, but their actual effects upon inhalation have yet to be determined.

Differentiation of black and white pitch (Burseraceae) oleoresins: A mass spectrometry-based chemoethnotaxonomic study.

ETHNOPHARMACOLOGICAL RELEVANCE: In the quilombola communities of the municipality of Oriximiná (Pará State, Brazil), Protium spp. (Burseraceae) oleoresins are distinguished in black and white pitch. White pitch oleoresins may be superior to black pitch in terms of quality, but the criteria used for their differentiation are fairly subjective. AIM OF THE STUDY: This study was designed to provide a scientific rationale for the traditional differentiation of black and white pitch oleoresins based on their non-volatile fraction. MATERIALS AND METHODS: Black and white pitch oleoresin samples collected in quilombola territories in Oriximiná were analysed by GC-EI-MS and UPLC-APCI-MS. The feasibility of EI and APCI mass spectrometry-based pattern recognition methods PLS-DA and Random Forest Analysis (RFA) for black and white pitch oleoresins differentiation was demonstrated. RESULTS: The UPLC-APCI-MS method allowed the separation of 43 triterpenoids. Assessment of the triterpenoid fingerprints by GC-EI-MS led to the tentative identification of ursa-9(11),12-dien-3-ol as a potential marker for black pitch oleoresins. PLS-DA and RFA applied to the APCI-MS and EI-MS data gave good models for black and white pitch oleoresins classification. The most important ions for the classifications of black pitch oleoresins by APCI-MS/PLS-DA and APCI-MS/RFA likely represented triterpenoid acids. CONCLUSIONS: The triterpenoid pattern differs between black and white pitch oleoresins. The characteristic presence of ursa-9(11),12-dien-3-ol and triterpenoids acids in black pitch oleoresins, along with other field observations, suggest that black pitch oleoresins are actually aged white pitch oleoresins.

Artemisia annua L. and photoresponse: from artemisinin accumulation, volatile profile and anatomical modifications to gene expression.

KEY MESSAGE: Blue and yellow light affected metabolism and the morphology. Blue and red promote the DOXP/MEP pathway. ADS gene expression was increased in plants cultivated under blue, promoting artemisinin content. Artemisinin-based combination therapies are the most effective treatment for highly lethal malaria. Artemisinin is produced in small quantities in the glandular trichomes of Artemisia annua L. Our aim was to evaluate the effect of light quality in A. annua cultivated in vitro under different light qualities, considering anatomical and morphological changes, the volatile composition, artemisinin content and the expression of two key enzymes for artemisinin biosynthesis. Yellow light is related to the increase in the number of glandular trichomes and this seemed to positively affect the molecular diversity in A. annua. Yellow light-stimulated glandular trichome frequency without triggered area enhancement, whereas blue light stimulated both parameters. Blue light enhanced the thickness of the leaf epidermis. The B-promoting effect was due to increased cell size and not to increased cell numbers. Green and yellow light positively influenced the volatile diversity in the plantlets. Nevertheless, blue and red light seemed to promote the DOXP/MEP pathway, while red light stimulates MVA pathway. Amorpha-4,11-diene synthase gene expression was significantly increased in plants cultivated under blue light, and not red light, promoting artemisinin content. Our results showed that light quality, more specifically blue and yellow light, positively affected secondary metabolism and the morphology of plantlets. It seemed that steps prior to the last one in the artemisinin biosynthesis pathway could be strongly influenced by blue light. Our work provides an alternative method to increase the amount of artemisinin production in A. annua without the use of transgenic plants, by the employment of blue light.

Biodegradation of Organophosphorus Compounds Predicted by Enzymatic Process Using Molecular Modelling and Observed in Soil Samples Through Analytical Techniques and Microbiological Analysis: A Comparison.

Organophosphorus compounds (OP) are chemicals widely used as pesticides in different applications such as agriculture and public health (vector control), and some of the highly toxic forms have been used as chemical weapons. After application of OPs in an environment, they persist for a period, suffering a degradation process where the biotic factors are considered the most relevant forms. However, to date, the biodegradation of OP compounds is not well understood. There are a plenty of structure-based biodegradation estimation methods, but none of them consider enzymatic interaction in predicting and better comprehending the differences in the fate of OPs in the environment. It is well known that enzymatic processes are the most relevant processes in biodegradation, and that hydrolysis is the main pathway in the natural elimination of OPs in soil samples. Due to this, we carried out theoretical studies in order to investigate the interactions of these OPs with a chosen enzyme-the phosphotriesterase. This one is characteristic of some soils' microorganisms, and has been identified as a key player in many biodegradation processes, thanks to its capability for fast hydrolyzing of different OPs. In parallel, we conducted an experiment using native soil in two conditions, sterilized and not sterilized, spiked with specific amounts of two OPs with similar structure-paraoxon-ethyl (PXN) and O-(4-nitrophenyl) O-ethyl methylphosphonate (NEMP). The amount of OP present in the samples and the appearance of characteristic hydrolysis products were periodically monitored for 40 days using analytical techniques. Moreover, the number of microorganisms present was obtained with plate cell count. Our theoretical results were similar to what was achieved in experimental analysis. Parameters calculated by enzymatic hydrolysis were better for PXN than for NEMP. In soil, PXN suffered a faster hydrolysis than NEMP, and the cell count for PXN was higher than for NEMP, highlighting the higher microbiological toxicity of the latter. All these results pointed out that theoretical study can offer a better comprehension of the possible mechanisms involved in real biodegradation processes, showing potential in exploring how biodegradation of OPs relates with enzymatic interactions.

Collection and identification of an unknown component from Eugenia uniflora essential oil exploiting a multidimensional preparative three-GC system employing apolar, mid-polar and ionic liquid stationary phases.

The present research deals with the collection and structural elucidation of an unknown component, accounting for about 35% of the essential oil obtained upon distillation of the leaves of Eugenia uniflora L., harvested during summer (January, 2017) in Paraná State (Southern Brazil). A multidimensional gas chromatographic preparative system, based on the coupling of three GC systems equipped with apolar, PEG and ionic liquid-based stationary phases, was successfully applied for the isolation of the chromatographic band relative to the unknown molecule. The use of wide-bore columns allowed for an increased sample capacity compared to conventional micro-bore columns, thus the injection of a neat sample was feasible, greatly reducing the total collection time. A higher chromatographic efficiency was afforded by the use of a multidimensional approach in the heart-cut mode, exploiting the different selectivity of three stationary phases, which ensured the attainment of a highly pure fraction. In only five runs, more than 3 milligrams were collected, with an average purity greater then 95%. Finally, the unknown component was subjected to nuclear magnetic resonance spectroscopy, mass spectrometry and condensed phase Fourier-transform infrared spectroscopy, leading to the identification of 6-ethenyl-6-methyl-3,5-di(prop-1-en-2-yl)cyclohex-2-en-1-one. The presented approach has been demonstrated to be effective for the isolation and structural elucidation of unknown molecules in complex samples, which will allow for further in-depth studies, like biological evaluation or pharmacological tests.

New approaches to monitor semi-volatile organic compounds released during coffee roasting using flow-through/active sampling and comprehensive two-dimensional gas chromatography.

A novel dynamic approach is described to profile volatile organic compound (VOC) and semi-VOC (SVOC) emission during coffee roasting aimed at analysing components present in the roasting plume, and to monitor their evolution during the process. Two sorbents - coconut shell charcoal (CSC) and styrene-divinylbenzene resin (XAD-2) - were evaluated while collecting substances in four sequential time intervals (0-3, 3-6, 6-9 and 9-12 min). Extracted VOCs (<200 Da) and SVOCs were analysed by gas chromatography (GC), and comprehensive two-dimensional gas chromatography (GC × GC) with flame ionisation (FID) and time-of-flight mass spectrometry (TOFMS) detection. Results showed CSC extraction presented poor recovery of VOCs and SVOCs released during roasting. However, XAD-2 was able to collect both groups, including SVOCs of >400 Da. GC × GC resolved many co-eluting compounds observed in 1D GC and allowed chemical group type cluster analysis, revealing that many non-polar VOCs are observed within the 0-3 min interval, and that the release of polar and higher molar mass SVOCs were mostly found within the 3-6 min interval. These group-type cluster analyses offer a broad spectrum chemical profile of the released substances. It may also reveal detailed insights into the roast process evolution over time.

Gas chromatography-mass spectrometry of sapucainha oil (Carpotroche brasiliensis) triacylglycerols comprising straight chain and cyclic fatty acids.

Sapucainha oil, which may be used to treat leprosy, comprises straight chain and cyclic fatty acids (FA), and triacylglycerols (TAG). The FA and TAG content of the oil sample was analysed using gas chromatography-electron ionisation mass spectrometry (GC-EIMS). FA analysis was performed after derivatisation to fatty acid methyl esters (FAME). For free FA and TAG analysis, the oil sample was dissolved in hexane and injected into a short, high-temperature column, for GC with MS analysis. Free FA and FAME were tentatively identified based on mass spectrum information of their molecular and fragment ions, as well as library matching. Overlapping TAG peaks were deconvoluted based on mass fingerprint data. The FA composition was utilised to predict possible TAG identities. FA residues of TAG were identified based on characteristic fragment ions, such as [M-RCO2]+, [RCO+128]+, [RCO+74]+ and RCO+ where R is the aliphatic hydrocarbon chain. FAME analysis showed that the cyclic FA hydnocarpic (36.1%), chaulmoogric (26.5%) and gorlic (23.6%) acids were the major components. In addition, straight chain FA such as palmitic, palmitoleic, stearic, oleic and linoleic acids were detected. Palmitic, oleic, hydnocarpic, chaulmoogric and gorlic acids were also detected as free FA in the oil sample. Six groups of TAG peaks were eluted from GC at temperatures ≥330 °C. After deconvolution and mass spectrum analysis, each TAG peak group was revealed to comprise 2 to 5 co-eluted TAG molecules; >18 TAG were identified. These TAG consisted of a mix of both cyclic and straight chain FA, but were mostly derived from cyclic FA.

Reduction of ßN-alkanoyl-5-hydroxytryptamides and diterpenes by yeast supplementation to green coffee during wet processing.

Coffee is one of the most consumed non-alcoholic beverages in the world. It is well known that some compounds present in coffee beans have important biological activities. In this study, evidence was turned to ßN-alkanoyl-5-hydroxytryptamides (C-5HTs) and to the furokaurane diterpenes cafestol and kahweol, associated with gastric irritation and increasing of blood cholesterol, respectively. Fermentation in coffee post-harvest wet process was induced by three Saccharomyces cerevisiae yeasts (for bakery, white and sparkling wines) as starter cultures. Variations in mass, time, temperature and pH (56 experiments under fractional factorial and mixture experimental designs) were tested. Substantial reductions for C-5HTs (up to 38% reduction for C20-5HT and 26% for C22-5HT) as well as for diterpenes (54% for cafestol and 53% for kahweol) were obtained after treating green coffee beans with 0.6 g of a 1:1:1 mixture the three yeasts for 12 h at 15 °C and pH 4. Caffeine and 5-CQA content, monitored in the green coffee beans, did not change. Therefore, the use of starter cultures during coffee post-harvest wet process has influence on the amount of some important compounds related to health and improves the sensory quality of the beverage.

Insecticidal activity of essential oils in controlling fall armyworm, Spodoptera frugiperda / Atividade inseticida de óleos essenciais para o controle da lagarta-do-cartucho do milho, Spodoptera frugiperda

Fall armyworm, Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) is one of the main pests in maize crop with developing resistance to chemical products and Bt technology. Therefore, alternative control methods such as essential oils are important steps in the implementation management strategies for this pest. This study aimed to evaluate the efficiency of essential oils (EOs) of Corymbia citriodora, Myrciaria dubia (Myrtaceae), Lippia microphylla (Verbenaceae) and Piper umbelattum (Piperaceae) in controlling S. frugiperda. The OEs were extracted and mortality tests were conducted with topic and volatile applications, in 30 second-instar caterpillars originated from insect rearing and artificial diet. As a control, we conducted tests with distilled water and acetone. EOs that provided mortality rates above 80% were submitted to chemical analysis for constituent identification. The efficient EOs were only those of C. citriodora and L. microphylla. For EO of C. citriodora, the LD80 was 7.06 ± 0.73 mg.g-1 in topical application and 5.85 ± 0.75 µL via volatile application. On the other hand, for EO of L. microphylla, DL80 was 9.95 ± 1.25 mg.g-1 in topical application and 18.56 ± 3.55 µL via volatile application. Chemical analysis showed that the main constituents were citronella for the EO of C. citriodora and (E)-caryophyllene and (E)-nerolidol to the EO of L. microphylla. EOs of C. citriodora and L. microphylla are promising for controlling S. frugiperda, with emphasis on the volatile effect of C. citriodora oil.(AU)

A lagarta-do-cartucho, Spodoptera frugiperda (Lepdoptera: Noctuidae), é uma das principais pragas na cultura do milho e nos últimos anos vem desenvolvendo resistência a produtos químicos e à tecnologia Bt. Métodos alternativos de controle, como o emprego de óleos essenciais, são um passo importante na implementação de estratégias de manejo para essa praga. O objetivo deste trabalho foi avaliar a eficiência dos óleos essenciais de Corymbia citriodora, Myrciaria dubia (Myrtaceae), Lippia microphylla (Verbenaceae) e Piper umbelattum (Piperaceae) no controle de S. frugiperda. Os óleos essenciais foram extraídos e testes de mortalidade com aplicação tópica e de voláteis foram conduzidos com 30 lagartas de segundo instar provenientes de criação massal e em dieta artificial. Como testemunha, foram conduzidos testes com água destilada e acetona. Os óleos essenciais que proporcionaram taxas de mortalidade acima de 80% foram submetidos à análise química para identificação de seus constituintes. Entre os óleos essenciais, mostraram-se eficientes apenas os de C. citriodora e L. microphylla. Para o óleo essencial de C. citriodora, a DL80 foi de 7,06 ± 0,73 mg.g-1 em aplicação tópica e 5,85 ± 0,75 µL via aplicação do volátil. Já para o óleo essencial de L. microphylla, a DL80 foi de 9,95 ± 1,25 mg.g-1 em aplicação tópica e 18,56 ± 3,55 µL via aplicação do volátil. A análise química demonstrou que os principais constituintes foram citronelal para o óleo essencial de C. citriodora, e (E)-cariofileno e (E)-nerolidol para o de L. microphylla. Os óleos essenciais de C. citriodora e L. microphylla são promissores para o controle de S. frugiperda, com destaque para o efeito volátil do óleo de C. citriodora.(AU)

Efficacy of essential oils from plants cultivated in the Amazonian Biome against gastrointestinal nematodes in sheep.

The excessive use of anthelmintics to control nematodes has resulted in anthelminthic resistance. Essential oils (EOs) are a rich source of bioactive molecules that can be assessed for their ability to control resistant parasite populations. The aims of this study were to screen EOs from 10 plant species in vitro for anthelmintic activity against Haemonchus contortus, evaluate the cytotoxicity of those EOs in a human immortalized keratinocyte cell line (HaCaT), and test the most promising EO candidate in vivo in Santa Inês sheep. The efficacy was investigated in vitro using an egg hatch test (EHT) and a larval development test (LDT). EO cytotoxicity was evaluated with the sulforhodamine-B assay. In the in vivo experiment, 28 Santa Inês sheep naturally infected were distributed into groups: G1-Mentha arvensis (EO5), 200 mg kg-1; G2-menthol, 160 mg kg-1; G3-negative control; and G4-positive control (monepantel). EO5, from M. arvensis (86.7% menthol), had the lowest LC50 and LC90 values in the EHT (0.10, 0.27 mg mL-1, respectively), good performance in the LDT (0.015, 0.072 mg mL-1, respectively), and the lowest cytotoxicity (190.9 µg mL-1) in HaCaT cells. In the in vivo test, a single dose of the EO5 (200 mg kg-1 BW) had an efficacy of approximately 50% on days 1, 14, and 21; however, values were not significantly from day 0. Conversely, pure menthol at a dose of 160 mg kg-1 BW showed no in vivo efficacy. This can be attributed to key factors related to bioavailability and pharmacology of terpenes in the host organism, as well as to the fact that menthol is mainly excreted as glucuronides in urine. Thus, further studies should be conducted with formulation systems that deliver bioactives directly to the abomasum, focusing on terpenes, whose excretion route is mainly via faeces.

Piper Essential Oils Inhibit Rhizopus oryzae Growth, Biofilm Formation, and Rhizopuspepsin Activity.

Piper is the largest genus of the Piperaceae family. The species of this genus have diverse biological activities and are used in pharmacopeia throughout the world. They are also used in folk medicine for treatment of many diseases in several countries including Brazil, China, India, Jamaica, and Mexico. In Brazil, Piper species are distributed throughout the national territory, making this genus a good candidate for biological activity screening. During our studies with Piper essential oils, we evaluated its activity against Rhizopus oryzae, the main agent of mucormycosis. The main compounds of seven Piper essential oils analyzed were Piper callosum-safrole (53.8%), P. aduncum-dillapiole (76.0%), P. hispidinervum-safrole (91.4%), P. marginatum-propiopiperone (13.2%), P. hispidum-γ-terpinene (30.9%), P. tuberculatum-(E)-caryophyllene (30.1%), and Piper sp.-linalool (14.6%). The minimum inhibitory concentration of Piper essential oils against R. oryzae ranged from 78.12 to >1250 µg/mL. The best result of total inhibition of biofilm formation was obtained with Piper sp. starting from 4.88 µg/mL. Considering the bioactive potential of EOs against planktonic cells and biofilm formation of R. oryzae could be of great interest for development of antimicrobials for therapeutic use in treatment of fungal infection.

Lipase-catalysed esters synthesis of cafestol and kahweol.

Cafestol and kahweol (C&K), two coffee diterpene alcohols with structural similarity which exhibit anticarcinogenic effects, were isolated from green coffee Arabica beans, followed by their lipase-catalysed esterification and purification by preparative high-performance liquid chromatography (HPLC). The isolation and enzymatic synthesis parameters of C&K esters were studied, with the latter optimised by a Central Composite Design; both procedures were monitored by gas chromatography. Scale up and improved isolation conditions resulted in 1.29 g of C&K, with 98% purity from 300 g of green Arabica beans. The highest C&K ester yields were observed using an alcohol:fatty acid molar ratio of 1:5, 73.3 mg mL-1 of CAL-B enzyme, 70 °C and 240 rpm for 3 days in toluene, leading to 85-88% conversion among a variety of tested C&K esters, including n-C14:0-C20:0, C18:1, C18:2 and C18:3.

Analysis of underivatised low volatility compounds by comprehensive two-dimensional gas chromatography with a short primary column.

Comprehensive two-dimensional gas chromatography (GC×GC) approaches with cryogenic modulation were developed for the qualitative analysis of selected low volatility compounds in raw coffee bean extracts, without derivatisation. The approaches employed short first (1D) and second (2D) dimension columns, specifically a 1D 65% phenyl methyl siloxane column (11m) and a 2D 5% phenyl methyl siloxane column (1m), which allowed elution of high molar mass compounds (e.g.>600Da). Solutes included hydrocarbons, fatty acids, diterpenes, tocopherols, sterols, diterpene esters, and di- and triacylglycerides. An oven temperature program up to 370°C was employed. The effects of experimental conditions were investigated, revealing that the GC×GC results strongly depended on the cryogenic trap T, and oven T program. An appropriate condition was selected and further applied for group type analysis of low volatility compounds in green Arabica coffee beans. Retention indices were compiled for 1D GC analysis and were similar for the composite column data in GC×GC. The elution of some compounds was confirmed by use of authentic standards. The approach allowed direct analysis of coffee extract in ethyl acetate solution, with improved analyte peak capacity (approximately 200 compounds were detected) without prior fractionation or pre-treatment of the sample. This avoided potential hydrolysis of high molar mass conjugate esters as well as degradation of thermally labile compounds such as the derivatives of the diterpenes cafestol and kahweol.