Analytical and chemometric strategies for elucidation of yerba mate composition.

Yerba mate, a popular plant consumed mainly as an infusion, possesses nutritional and medicinal properties attributed to its secondary metabolites. This study aimed to develop strategies to elucidate the phenolic composition of yerba mate samples from Brazil, Argentina, Uruguay, and Paraguay. Optimization of ultrasonic-assisted extraction (UAE) was performed, and the extracted compounds were characterized using ultra-high-pressure liquid chromatography coupled with quadrupole/time-of-flight mass spectrometry (UHPLC-QTOF-MS), molecular fluorescence and high-pressure liquid chromatography with diode-array detection (HPLC-DAD). Chemometric analysis, including parallel factor analysis (PARAFAC) and principal component analysis (PCA) explored metabolite profiles and identify patterns. PARAFAC modelling of the molecular fluorescence results revealed higher pigment content in Brazilian samples, while other countries' samples exhibited higher phenolic content. PCA modeling of HPLC-DAD results indicated that cultivated yerba mate contained higher chlorogenic acids levels, and samples from Argentina, Paraguay, and Uruguay exhibited higher concentrations of chlorogenic acids and flavonoids.

Green-based methods to obtain bioactive compounds from Ilex guayusa Loes. using polar solvent.

Many biological activities have been reported for the Ilex genus. However, few studies in the literature have reported on guayusa. To address this gap in our knowledge, chemical analysis of guayusa leaves was made. Extracts were obtained by applying Soxhlet, maceration, supercritical CO2 and pressurised liquid extraction techniques, using water and ethanol as solvent/cosolvent. Extracts were evaluated for their phenolic content and antioxidant capacity. The chemical profile was obtained from HPLC. In raw guayusa leaves were identified caffeine (2.27 ± 0.05%), protein (15.31 ± 0.07%) and lipids (11.81 ± 0.14%). Extracts presented the highest phenolic content (156.56 ± 1.32 mg GAE g-1) and the best antioxidant activity (EC50= 61.85 ± 0.21 µg mL-1) when water was used as solvent/cosolvent. Through HPLC, three main substances were determined and quantified in the extracts: caffeine, theobromine and 5-caffeoylquinic acid. Based on these results, guayusa may be considered a natural source of compounds with potential application in the food and pharmaceutical industries.

Production of volatile compounds by yeasts using hydrolysed grape seed oil obtained by immobilized lipases in continuous packed-bed reactors.

Lipases CAL-B, TLL, and RML were used in the synthesis of free fatty acids of grape seed oil as heterogeneous substrate. The best enzyme was used to optimize the reaction variables temperature, enzyme content, and molar ratio of water:oil in batch reactions using experimental planning. The ideal conditions to produce free fatty acids using pure RML were 45 °C, 12:1 substrate molar ratio, and 15% enzyme, resulting in 66% of oil hydrolysis and a productivity of 0.54 mol L-1 min-1 in 4 h of reaction at 180 rpm. Repeated batches of reaction were performed testing the operational stability of RML, results showing that this enzyme could be used for at least 20 cycles keeping more than 80% of its initial activity, suggesting its potential use in industrial processes. The synthesis of free fatty acids was then evaluated in continuous reactions using packed-bed reactor (PBR). The highest productivity in the continuous process was 6.85 mol L-1 min-1, using only RML, showing an operational stability higher than 80% of its initial conversion capacity after 11 days of operation, at a flow rate of 0.13 mL min-1 at 45 °C. We evaluated the use of this hydrolyzed oil as substrate for lactone bioproduction using Galactomyces geotrichum UFMG-CM-Y3276, G. geotrichum UFMG-CM-Y3558, and Geotrichum klebahnii UFMG-CM-Y3014 screened for their oil-hydrolysis ability. Volatile compounds were qualitatively identified in GC-MS as γ-octalactone and γ-nonalactone.

Wine lees from the 1st and 2nd rackings: valuable by-products.

Phenolic compounds of the first and second racking wine lees, including anthocyanins, were qualitatively and quantitatively analyzed by HPLC-DAD-MS. Wine lees from both rackings displayed similar chromatographic profiles. Therefore, it was impossible to differentiate the qualitative results regarding phenolic compounds. On the other hand, those from the second racking presented, on average, concentration of polyphenols twice as high. While the ones from the first racking displayed ca. 1600 mg phenolic compounds and 400 mg anthocyanins per kg of dry matter, those from the second racking have shown ca. 3300 mg phenolic compounds and 700 mg anthocyanins per kg of dry matter. These outcomes indicate that, although the wine lees from the first racking can be employed as a resource for phenolic compounds recovery, those from the second racking are more appropriate for this purpose.

GC×GC/qMS analyses of Campomanesia guazumifolia (Cambess.) O. Berg essential oils and their antioxidant and antimicrobial activity.

The present study investigated the essential oil obtained from Campomanesia guazumifolia (Cambess.) O. Berg, an aromatic plant used in Brazilian folk medicine. The chemical composition was performed by GC×GC/qMS. The antioxidant and antimicrobial activities were evaluated by DPPH and BCB and, MIC assays, respectively. Sixty-eight compounds were identified in the oil, where the major compounds were bicyclogermacrene (15%), globulol (5%) and spathulenol (5%). Sesquiterpene hydrocarbons (29 compounds) and oxygenated sesquiterpenes (20 compounds) were the most representative classes of terpenes. DPPH (IC50 value 26.1 ± 0.5 µg/mL) and BCB (68.3 ± 1.5%) values indicated a significant antioxidant activity. The essential oil strongly inhibited Staphylococcus aureus (MIC 15 ± 0.1 µg/mL), Escherichia coli (MIC 25 ± 0.2 µg/mL) and Candida albicans (MIC 5 ± 0.1 µg/mL). The results give a deeper understanding of the chemical composition and report for the first time the antioxidant and antimicrobial potential of the C. guazumifolia essential oil.

Do we truly understand pollination syndromes in Petunia as much as we suppose?

Petunia is endemic to South America grasslands; member of this genus exhibit variation in flower colour and shape, attracting bees, hawkmoths or hummingbirds. This group of plants is thus an excellent model system for evolutionary studies of diversification associated with pollinator shifts. Our aims were to identify the legitimate pollinator of Petunia secreta, a rare and endemic species, and to assess the importance of floral traits in pollinator attraction in this Petunia species. To determine the legitimate pollinator, field observations were conducted, and all floral visitors were recorded and evaluated. We also measured the nectar volume and sugar concentration. To characterize morphological cues for pollinators, we assessed the ultraviolet (UV)-light response in detached flowers, and characterized the floral pigments and pollen volatile scents for four different Petunia species that present different pollination syndromes. Petunia secreta shares the most recent ancestor with a white hawkmoth-pollinated species, P. axillaris, but presents flavonols and anthocyanin pigments responsible for the pink corolla colour and UV-light responses that are common to bee-pollinated Petunia species. Our study showed that a solitary bee in the genus Pseudagapostemon was the most frequent pollinator of P. secreta, and these bees collect only pollen as a reward. Despite being mainly bee-pollinated, different functional groups of pollinators visit P. secreta. Nectar volume, sugar concentration per flower, morphology and components of pollen scent would appear to be attractive to several different pollinator groups. Notably, the corolla includes a narrow tube with nectar at its base that cannot be reached by Pseudagapostemon, and flowers of P. secreta appear to follow an evolutionary transition, with traits attractive to several functional groups of pollinators. Additionally, the present study shows that differences in the volatiles of pollen scent are relevant for plant mutualistic and antagonist interactions in Petunia species and that pollen scent profile plays a key role in characterizing pollination syndromes.

Chemical characterisation of Piper amalago (Piperaceae) essential oil by comprehensive two-dimensional gas chromatography coupled with rapid-scanning quadrupole mass spectrometry (GC×GC/qMS) and their antilithiasic activity and acute toxicity.

INTRODUCTION: Piper amalago has a distribution from Mexico to Brazil; their aerial parts have been used in folk medicine to treat diuretic and kidney diseases. OBJECTIVE: The purpose of this study was to obtain a deeper understanding of the chemical composition of essential oils (EOs) extracted from both the leaves and stems of P. amalago, compare them, and evaluate their antilithiasic activity and acute toxicity. METHODOLOGY: Extraction was performed by hydrodistillation, whereas chemical characterisation by two-dimensional gas chromatography coupled with rapid-scanning quadrupole mass spectrometry (GC×GC/qMS). The antilithiasic activity was evaluated by the effect of the EOs on calcium oxalate crystallisation in vitro. The turbidity index and the number of crystals formed were determined and used as an estimative of the activity. In the acute toxicity assay, the effects of a single oral dose of the EOs in Wistar rats were determined. General behaviour, adverse effects, and mortality were determined. RESULTS: A total of 322 compounds were identified in the EOs. The sesquiterpenes displayed the highest contribution in leaves EOs among which included bicyclogermacrene and δ-cadinene. Sesquiterpenes and oxygenated sesquiterpenes displayed the highest contribution in EOs from stems, among which included bicyclogermacrene and α-cadinol. The EOs demonstrated an excellent action on the crystals growth inhibition, and the oral dose tested did not induce significant changes in the parameters for acute toxicity. CONCLUSION: The oils have a high chemical complexity, and there are differences between their compositions, which could explain the observed differences in antilithiasic activity. The findings support the use of this plant in folk medicine to treat kidney diseases.

Quantification of nitrogen compounds in diesel fuel samples by comprehensive two-dimensional gas chromatography coupled with quadrupole mass spectrometry.

Although several methods for the analysis of nitrogen compounds in diesel fuel have been described in the literature, the demand for rapid, sensitive, and robust analyses has increased in recent years. In this study, a comprehensive two-dimensional gas chromatographic method was developed for the identification and quantification of nitrogen compounds in diesel fuel samples. The quantification was performed using the standard addition method and the analysis was conducted using comprehensive two-dimensional gas chromatography coupled with fast quadrupole mass spectrometry. This study is the first to report quantification of nitrogen compounds in diesel fuel samples using the standard addition method without fractionation. This type of analysis was previously performed using many laborious separation steps, which can lead to errors and losses. The proposed method shows good linearity for target nitrogen compounds evaluated (m-toluidine, 4-ethylaniline, indole, 7-methylindole, 7-ethylindole, carbazole, isoquinoline, 4-methylquinoline, benzo[h]quinolone, and acridine) over a range from 0.05 to 2.0 mg/L, and limits of detection and quantification of <0.06 and 0.16 mg/L, respectively, for all nitrogen compounds studied.

Comprehensive two dimensional gas chromatography with fast-quadrupole mass spectrometry detector analysis of polar compounds extracted from the bio-oil from the pyrolysis of sawdust.

In this paper it is studied the most polar fractions of bio-oil produced by fast pyrolysis of Lignocel BK40-90 (sawdust from forest timber). The biomass was submitted to the pyrolysis in an existing FCC pilot plant that was adapted for this procedure. The equipment consists of a fluidized bed reactor with nitrogen injection. The unit operates with continuous biomass feeding and continuous solids circulation. The produced bio-oil was submitted to an aqueous alkaline extraction, isolating the acidic compounds that were analyzed by one-dimensional gas chromatography and comprehensive two-dimensional gas chromatography with quadrupole mass spectrometry detection (qMS). One hundred and thirty compounds (mainly phenols and ketones) were tentatively identified in the extract, some of them by the use of retention indexes. The main differences between chromatographic techniques were the substantial increasing in the peak capacity of GC×GC and the resolution of some co-elutions that occurred in GC/qMS. It is also possible to conclude that this extract is rich in important raw materials for the chemical industry and can be used for this end.

Characterization of the yerba mate (Ilex paraguariensis) volatile fraction using solid-phase microextraction-comprehensive 2-D GC-MS.

The present research is focused on the use of a solid-phase microextraction-comprehensive 2-D GC methodology, in the analysis of the volatile fraction of yerba mate. Yerba mate is used for the generation of a tea-like beverage, widely consumed in South America. A rapid-scanning quadrupole mass spectrometer (qMS), employed as a detection system and operated at a 25 Hz scanning frequency, supplied high-quality mass spectra. The effectiveness of the 3-D comprehensive 2-D GC-qMS experiment was compared to that of GC-qMS analysis on the same sample. Peak identification, in both applications, was achieved through MS library matching, with the support of linear retention index data. Apart from a great increase in the number of analytes separated (approx. by a factor of 5) and identified (approx. by a factor of 3.5), the comprehensive 2-D GC-qMS approach enabled the determination of a high number of hazardous contaminants (aliphatic hydrocarbons, polycyclic aromatic hydrocarbons, and plasticizers), barely visible in the GC-qMS analysis.

High efficiency liquid chromatography techniques coupled to mass spectrometry for the characterization of mate extracts.

There is growing interest related to rapid screening and full characterization of the constituents of plants with medicinal properties; among these, "Mate" or Yerba Maté is a tea-like beverage widely consumed in South America, obtained from the dried leaves of Ilex paraguariensis. The high content in polyphenols accounts for in vitro and in vivo antioxidant activity of the extracts obtained from this plant; on the other hand, the high complexity of the samples extracted, depending on the method employed, may preclude complete resolution by conventional HPLC techniques. For this purpose, a comprehensive two-dimensional liquid chromatography (LC x LC) system, comprised of an RP-Amide first dimension and a partially porous octadecylsilica column in the second dimension, has been compared with a one-dimensional system. The latter was operated using a partially porous octadecylsilica column, with diode array (DAD) and electrospray/ion trap-time of flight (ESI/IT-TOF) detection for the most complex extracts. The employment of the hybrid mass spectrometer allowed unequivocal identification of several compounds in the mate extracts. Using LC x LC-MS(3), it was possible to discriminate between congeners of chlorogenic acids, along with monoacyl- and diacylchlorogenic acid esters.

Pressurized liquid extraction of mate tea leaves.

The objective of this work is to investigate the influence of process parameters on the pressurized liquid extraction (PLE) of Ilex paraguariensis leaves. A factorial 2(6-2) experimental design was employed using responses as the extraction yield and the chromatographic profile of the extracts. The extraction time, polarity of solvent, amount of sample, numbers of PLE cycles, flushing volume and extraction temperature were selected as independent variables (factors). Results obtained indicated that the solvent polarity was the most significant variable in the study, while the amount of sample and extraction temperature also showed significant effect. The other variables did not present significant influence in the yield of extraction. GC/MS analysis of the extract enabled the identification of saturated hydrocarbons, fatty acids, fatty acid methyl esters, phytosterols and theobromine in the extracts. Quantitative analysis of four compounds presented in the extracts (caffeine, phytol, vitamin E and squalene) was performed by the GC/MS in the SIM mode.

Extraction of grape seed oil using compressed carbon dioxide and propane: extraction yields and characterization of free glycerol compounds.

The main objective of this work was to compare the extraction of grape seed oil with compressed carbon dioxide and propane on the extraction yields and chemical characteristics of free glycerol compounds. The experiments were performed in a laboratory scale unit in the temperature range of 30 to 60 degrees C and pressures from 60 to 254 bar. The results showed that propane is a more suitable solvent for grape seed oil extraction than carbon dioxide, as higher extractions yields and a very fast kinetic of extraction were achieved with this solvent. In relation to compressed carbon dioxide extractions, both temperature and density presented a very pronounced and positive effect on the extraction yield. The oils extracted were analyzed qualitatively and quantitatively with regard to the free glycerol compounds, mainly fatty acids, ethyl, and methyl esters. The results showed that these compounds are present in low concentration in vegetable oil (<3%) and that, in general, samples extracted with propane present a smaller amount of peaks of free glycerol compounds in the oil than samples extracted with carbon dioxide.

Pressurized liquid extraction of vitamin E from Brazilian grape seed oil.

The goal of this paper is to optimize the pressurized liquid extraction (PLE) of vitamin E from grape seed oil from residues of the wine industry. For this purpose an experimental planning to optimize the extraction of Brazilian grape seed oil by means of PLE with hexane as solvent was applied and the results are compared with conventional methods (Soxhlet and mechanical press extraction). Vitamin E was separated and analyzed using HPLC with UV detection. This study demonstrates the ability of the PLE in extracting grape seed oil rich in vitamin E.

Influence of drying methods and agronomic variables on the chemical composition of mate tea leaves (Ilex paraguariensis A. St.-Hil) obtained from high-pressure CO2 extraction.

The main objective of this work is to assess the influence of two drying methods (microwave and vacuum oven) and some agronomic variables (plant fertilization conditions and sunlight intensity) on the characteristics of mate tea (Ilex paraguariensis) leaves extracts obtained from high-pressure carbon dioxide extractions performed in the temperature range from 20 to 40 degrees C and from 100 to 250 bar. Samples of mate were collected in an experiment conducted under agronomic control at Ervateira Barão LTDA, Brazil. Chemical distribution of the extracts was evaluated by gas chromatography coupled with a mass spectrometer detector (GC/MS). In addition to extraction variables, results showed that both sample drying methods and agronomic conditions exert a pronounced influence on the extraction yield and on the chemical distribution of the extracts.

Influence of agronomic variables on the macronutrient and micronutrient contents and thermal behavior of mate tea leaves (Ilex paraguariensis).

The influence of agronomic variables (light intensity, age of leaves, and fertilization type) on the content of macronutrients and micronutrients (potassium, calcium, sodium, magnesium, manganese, iron, zinc, and copper) of tea leaves was assessed by acid digestion, followed by flame atomic absorption spectrometry (FAAS). The thermal behavior of mate tea leaves (Ilex paraguariensis) was also studied in this work. Samples of mate (Ilex paraguariensis) were collected in an experiment conducted under agronomic control at Erva-Mate Barão Commerce and Industry LTD (Brazil). The results showed that the mineral content in mate is affected by the agronomic variables investigated. In general, the content of mineral compounds analyzed is higher for younger leaves and for plants cultivated in shadow. Thermal analysis of samples indicated a similar behavior, with three typical steps of decomposition: loss of water, degradation of low-molecular weight compounds, and degradation of residual materials.

The use of ultrasound in the extraction of Ilex paraguariensis leaves: a comparison with maceration.

The objective of this work is to discuss the main parameters that influence the sonication extraction of Ilex paraguariensis leaves. For this purpose, the extraction time, solvent polarity, solvent volume, sample mass and particle size were evaluated. Results showed that the main variable affecting the extraction process was the solvent polarity. Though in a less extent, temperature and extraction time also demonstrated to be important parameters, while the other variables did not present a significant influence on the extraction yield. The extracts at the optimized condition were compared with those obtained by maceration, in terms of mass yield and chemical composition. The major compounds identified in the extracts were caffeine and palmitic acid. Some saturated hydrocarbons such as fatty acids, fatty acid methyl esters, phytosterols, and theobromine were also identified in the fractions.

Comparison of soxhlet, ultrasound-assisted and pressurized liquid extraction of terpenes, fatty acids and Vitamin E from Piper gaudichaudianum Kunth.

This paper describes a comparative study of extraction methods of terpenes (terpenic alcohols and phytosterols), fatty acids and Vitamin E from leaves of Piper gaudichaudianum Kunth. The analysis of extracts was done by gas chromatography with mass spectrometric detection. The identification and quantification was made by co-injections of the extract with certified standards. The use of pressurized liquid extraction (PLE; Dionex trade name: ASE, for accelerated solvent extraction) decrease significantly the total time of extraction, the amount of solvent and the manipulation of sample and solvents in comparison with soxhlet (SE) and ultrasound-assisted (USE). In addition, PLE was more effective for the extractions of terpenes (terpenic alcohols and phytosterols), fatty acids and Vitamin E.

Optimization of pressurized liquid extraction of Piper gaudichaudianum Kunth leaves.

Piperaceae family is original from tropical regions and it shows more than 700 species around the world. Piper gaudichaudianum Kunth is the specie more abundant in Brazil, occurring from Northeast to South Brazil. In this paper, it was investigated the influence of some experimental parameters on the pressurized liquid extraction (PLE) of P. gaudichaudianum Kunth leaves, using petroleum ether as extractor solvent. The optimization of the main variables involved in the PLE process (extraction temperature and time) has been done by response surface methodology (RSM) using, as responses, the extraction yield and the chromatographic profile (GC/MS) of the extracts. The optimized procedure employed 3 g of ground leaves, 10 min of extraction and one cycle of extraction at 85 degrees C. The major compounds present in the petroleum ether extracts were: palmitic acid, stearic acid and nerolidol. The results presented in this work show the possibility of using a fast and easy process to recover compounds from P. gaudichaudianum Kunth.

Chemical composition of mate tea leaves (Ilex paraguariensis): a study of extraction methods.

The objective of this work was to investigate the extraction of Ilex paraguariensis leaves by means of three extraction techniques: pressurized liquid extraction (PLE, also called accelerated solvent extraction--ASE), maceration, and sonication. Samples of mate tea leaves were collected from an experiment conducted under agronomic control at Indfistria e Comércio de Erva-Mate Barão LTDA, Brazil. Six solvents with increasing polarities (n-hexane, toluene, dichloromethane, ethyl acetate, acetone, and methanol) were used in this investigation. Chemical analysis of the extracts was performed by GC coupled with a mass spectrometer detector. The identification and quantification were accomplished by coinjections of certified standards. The results showed that no significant differences in the qualities of the extracts were noticed regarding the extraction methods. On the other hand, the PLE technique was found to be more effective for the extractions of caffeine, phytol, palmitic, and stearic acid. The use of PLE led to a significant decrease in the total extraction time, amount of solvent consumption, and manipulation of samples compared to maceration and ultrasound-assisted extraction methods.