Profiling of seized Cannabis sativa L. flowering tops by means of microwave-assisted hydro distillation and gas chromatography analyses.

This research aimed to support police forces in their battle against illicit drug trafficking by means of a multi-technique approach, based on gas chromatography. In detail, this study was focused on the profiling of volatile substances in narcotic Cannabis sativa L. flowering tops. For this purpose, the Scientific Investigation Department, RIS Carabinieri of Messina, provided 25 seized samples of Cannabis sativa L. The content of Δ9-tetrahydrocannabinol (THC), useful to classify cannabis plant as hemp (≤ 0.2 %) or as marijuana (> 0.2 %), was investigated. Essential oils of illicit drug samples were extracted using a microwave-assisted hydro-distillation (MAHD) system; GC-MS and GC-FID analytical techniques were used for the characterization of the terpenes and terpenoids fingerprint. Furthermore, the enantiomeric and carbon isotopic ratios of selected chiral compounds were investigated using a heart-cutting multidimensional GC (MDGC) approach. The latter exploited a combination of an apolar column in the first dimension, and a chiral cyclodextrin-based column in the second one, prior to parallel isotope-ratio mass spectrometry (C-IRMS) and MS detection. Finally, all the data were gathered into a statistical model, to demonstrate the existence of useful parameters to be used for the classification of seized samples.

Measurement of short-chain fatty acids in human plasma by means of fast gas chromatography-mass spectrometry.

A rapid and practicable analytical method for the measurement of short-chain fatty acids (SCFAs) in human plasma was developed. The extraction procedure involved the use of acidified water and methyl tert-butyl ether (MTBE), while the separation and detection of SCFAs, including acetic, propionic, and butyric acids was carried out by using gas chromatography-mass spectrometry (GC-MS) technique. The novelty of the research involves reducing the analysis time (less than 7 min) by using the novel fast GC-MS method. A narrow-bore GC capillary column of dimensions 30 m × 0.25 mm ID × 0.25 µm df with acid-modified poly(ethylene glycol) stationary phase was employed for the chromatographic separation. The signals of target compounds were acquired in selected ion monitoring (SIM) mode monitoring a quantifier ion (Q) and two qualifier ions (q1 and q2). Linearity of the method, limits of detection (LoD) and quantification (LoQ) were evaluated. In detail, regression coefficients of the calibration curves were between 0.9960 and 0.9933; LoDs ranged from 0.02 µM to 0.03 µM, while LoQs from 0.06 µM to 0.10 µM.

Fast gas chromatography-tandem mass spectrometry under milder electron ionization conditions for the assay of vitamin D metabolites in human serum.

The proposed research was focused on the development of a gas chromatography-tandem mass spectrometry (GC-QqQ-MS) method under milder electron ionization (EI) conditions for the assay of vitamin D metabolites in human serum. Efficiency of three different silylation agents was evaluated for the conversion of vitamin D species into trimethylsilyl (TMS) derivatives, among which N-methyl-N-(trimethylsilyl)-trifluoroacetamide (MSTFA) proved to be the most effective. Indeed, the MSTFA reagent was able to convert in TMS ether even the 25-hydroxyl vitamin D derivative that, as known, possesses steric hindrance problems. The separation of vitamin D compounds was obtained in about 11.5 min using a narrow-bore column of dimensions 30 m × 0.25 mm ID × 0.10 µm df with a poly(5% diphenyl/95% dimethyl siloxane) stationary phase. The mass spectrometry ionization of the silylated derivatives was performed under milder EI conditions (20-eV energy) that, respect to common 70-eV energy, generated scan mass spectra with higher relative and absolute intensities of high-mass diagnostic ions, along with a reduced abundance of the low-mass. The signals of the ionized compounds were acquired in multi-reaction-monitoring (MRM) mode, thus enabling the obtainment of highly-sensitive and selective quantitative data. The developed method was validated in term of linearity, accuracy, limits of detection (LoD) and quantification (LoQ). In detail, regression coefficients of the calibration curves were between 0.9959 and 0.9999; LoDs ranged from 0.06 ng mL-1 to 0.73 ng mL-1 and LoQs from 0.16 ng mL-1 to 2.45 ng mL-1. With respect to accuracy, the serum SRM 972a certified reference material (Vitamin D metabolites in frozen human serum) (Levels 1-4) was analyzed.

Flowering phenophases influence the antibacterial and anti-biofilm effects of Thymus vulgaris L. essential oil.

BACKGROUND: Essential oils are becoming increasingly popular in medicinal applications because of their antimicrobial effect. Thymus vulgaris L. (Lamiaceae) is a well-known and widely cultivated medicinal plant, which is used as a remedy for cold, cough and gastrointestinal symptoms. Essential oil content of thyme is responsible for its antimicrobial activity, however, it has been reported that the chemical composition of essential oils influences its biological activity. In order to explore flowering phenophases influence on the chemical composition of thyme essential oil and its antibacterial and anti-biofilm activity, plant materials were collected at the beginning of flowering, in full bloom and at the end of flowering periods in 2019. METHODS: Essential oils from fresh and dried plant materials were distilled and analyzed with gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detection (GC-FID). The antibacterial activity was performed by broth microdilution and thin layer chromatography-direct bioautography (TLC-DB) assays and the anti-biofilm effect by crystal violet assay, respectively. Scanning electron microscopy was applied to illustrate the cellular changes of bacterial cells after essential oil treatment. RESULTS: Thymol (52.33-62.46%) was the main component in the thyme essential oils. Thyme oil distilled from fresh plant material and collected at the beginning of flowering period exerted the highest antibacterial and anti-biofilm activity against Haemophilus influenzae, H. parainfluenzae and Pseudomonas aeruginosa. CONCLUSION: The different flowering periods of Thymus vulgaris influence the antibacterial and anti-biofilm activity of its essential oils, therefore, the collection time has to be taken into consideration and not only the full bloom, but the beginning of flowering period may provide biological active thyme essential oil.

Melissa officinalis L. as a Sprout Suppressor in Solanum tuberosum L. and an Alternative to Synthetic Pesticides.

The goal of this research was to screen plant essential oils (EOs) as sprout inhibitors or suppressors in potato (Solanum tuberosum L.). Three controlled environment experiments were conducted to screen 18 EOs and several pure compounds as sprout inhibitors. The EOs were applied using the wicked method on potato cv. Gala in 19 L plastic containers. The results indicated that Melissa officinalis L. EO inhibited sprouting, while Coriandrum sativum L. seed oil and the EO blend of Lavandula angustifolia Mill. and Salvia sclarea L. suppressed sprouting. The EOs of interest were analyzed using gas chromatography coupled to mass spectrometry (GC-MS) and/or a flame ionization detector (GC-FID); the detailed chemical profiles are provided. The M. officinalis EO was fractionated into seven fractions, and these were tested on minitubers. We identified two fractions (F and A) that suppressed potato sprouting better than the whole oil. The GC-MS-FID analyses of M. officinalis EO fraction A identified myrcene, Z-ocimene, E-ocimene, trans-caryophyllene, and α-humulene as the main constituents, while the main constituents of fraction F were α-terpineol, ß-citronellol, and geraniol. The pure isolated compounds, together with the major compound in M. officinalis EO (citral), were tested for sprout suppression on three potato cultivars (Ranger Russet, Terra Rosa, and Dakota TrailBlazer), which revealed that ß-citronellol reduced the sprout length and the number of sprouts in all three cultivars, while citral and (+)-α-terpineol reduced the sprout length and the number of sprouts in Ranger Russet relative to the two controls in all three cultivars. Myrcene had a stimulating effect on the number of sprouts in Cv. Terra Rosa. However, none of the pure compounds suppressed sprouting completely or were comparable to the EO of M. officinalis.

Anti-inflammatory effects of lavender and eucalyptus essential oils on the in vitro cell culture model of bladder pain syndrome using T24 cells.

BACKGROUND: Interstitial cystitis (IC) has a chronic chemical irritation and inflammation of non-bacterial origin in the bladder wall leading to various severe symptoms. There is evidence that chronic inflammation is significantly associated with abnormal urothelial barrier function, epithelial dysfunction. This is the underlying cause of urothelial apoptosis and sterile inflammation. METHOD: The anti-inflammatory effects of lavender and eucalyptus essential oils (EOs) and their main components (linalool and eucalyptol) were investigated in the T24 human bladder epithelial cell line on TNFα stimulated inflammation, at 3 types of treatment schedule. The mRNA of pro-inflammatory cytokines (IL-1ß, IL-6, IL-8) were measured by Real Time PCR. Human IL-8 ELISA measurement was performed as well at 3 types of treatment schedule. The effects of lavender and eucalyptus EOs and their main components were compared to the response to NFκB inhibitor ACHP (2-amino-6-[2-(cyclopropylmethoxy)-6-hydroxyphenyl]-4-(4-piperidinyl)-3-pyridinecarbonitrile). RESULT: There is no significant difference statistically, but measurements show that lavender EOs are more effective than eucalyptus EO. Long time treatment (24 h) of both lavender EO and linalool showed higher effect in decreasing pro-inflammatory cytokine mRNA expression than ACHP inhibitor following TNFα pre-treatment. Moreover, both lavender EOs were found to be significantly more effective in decreasing IL-8 secretion of T24 cells after TNFα pre-treatment compared to the ACHP NFκB-inhibitor. CONCLUSION: The lavender EOs may be suitable for use as an adjunct to intravesical therapy of IC. Their anti-inflammatory effect could well complement glycosaminoglycan-regenerative therapy in the urinary bladder after appropriate pharmaceutical formulation.

Simultaneous evaluation of the enantiomeric and carbon isotopic ratios of Cannabis sativa L. essential oils by multidimensional gas chromatography.

Recent times have witnessed an upsurge of interest in hemp and hemp-derived products, as driven by the scientific findings specific to the pharmacological properties of Cannabis sativa L. and its constituents. There has been evidence that the terpene profile, along with the cannabinoid content, produces in humans the effects associated with different strains, beyond fragrance perception. A great deal of effort has been put into developing analytical approaches to strengthen the scientific knowledge on cannabis essential oil composition and provide effective tools for ascertaining the authenticity of commercial cannabis samples. For this concern, enantio-selective-GC-C-IRMS has proven to be effective for assessing the ranges characteristic of the genuine samples and detecting any fraudulent additions. This research aimed at providing for the first time the enantiomeric and isotopic ratios of target terpenes in cannabis essential oils, obtained from microwave-assisted hydro-distillation from the fresh and dried inflorescences of different cannabis varieties. Implementing multidimensional gas chromatography separation was mandatory prior to detection, in order to obtain accurate δ13C values and enantiomeric data from completely separated peaks. For this purpose, a heart-cut method was developed, based on the coupling of an apolar first dimension column to a secondary chiral cyclodextrin-based stationary phase. Afterwards, the data gathered from enantio-selective-MDGC-C-IRMS/qMS analysis of a set of genuine samples were used to evaluate the quality of nineteen commercial cannabis essential oils purchased from local stores. Remarkably, the data in some cases evidenced enantiomeric ratios and δ13C values outside the typical ranges of genuine oils. Such findings suggest the usefulness of the method developed to ascertain the genuineness and quality of cannabis essential oils.

Anti-inflammatory effect of lavender (Lavandula angustifolia Mill.) essential oil prepared during different plant phenophases on THP-1 macrophages.

BACKGROUND: Pseudomonas aeruginosa is the most common Gram-negative bacterium associated with nosocomial respiratory infections. Lavender essential oil is mainly used in aromatherapy, but it has several pharmacological and therapeutic properties. Furthermore, it possesses antifungal and antibacterial activities. The anti-inflammatory activity of essential oils may depend on the composition and the ratio of the compounds. The constitution of the essential oils extracted from the different stages of flowering period varies, which makes it plausible that the collection time of the flowers influences the anti-inflammatory effects. Different types of essential oils reduce inflammation acting similarly by modulating the activity and action of the NFκB signalling pathway, which is the major regulator of the transcription of pro-inflammatory cytokines. METHODS: Lavender essential oils were distilled from lavender plant cultivated in Hungary and the flowers were harvested at the beginning and at the end of flowering period. The experiments were carried out on THP-1 human monocyte/macrophage cell line as in vitro cell culture model for monitoring the effects of lavender essential oils and the main compound linalool on P. aeruginosa LPS stimulated inflammation. The mRNA and protein levels of four pro-inflammatory cytokines, IL-6, IL-1ß, IL-8 and TNFα were determined by Real Time PCR and ELISA measurements. The effects of essential oils were compared to the response to two NFκB inhibitors, luteolin and ACHP. RESULTS: Linalool and lavender essential oil extracted from plants at the beginning of flowering period were successful in decreasing pro-inflammatory cytokine production following LPS pretreatment. In case of IL-8 and IL-1ß lavender oil showed stronger effect compared to linalool and both of them acted similarly to NFκB inhibitors. Pretreatments with linalool and lavender essential oil/beginning of flowering period prevented pro-inflammatory cytokine production compared to LPS treatment alone. Although lavender essential oil/end of flowering period decreased IL-6, IL-1ß and IL-8 mRNA expression in case of LPS pretreatment, it was not capable to reduce cytokine secretion. CONCLUSION: Based on our results it has been proven that lavender essential oil extracted at the beginning of flowering period is a potent inhibitor of the synthesis of four pro-inflammatory cytokines IL-6, IL-8, IL-ß and TNFα of THP-1 cells. This supports the relevance of the collection of the lavender flowers from early blooming period for essential oil production and for the utilization as an anti-inflammatory treatment.

Development of a Novel Microwave Distillation Technique for the Isolation of Cannabis sativa L. Essential Oil and Gas Chromatography Analyses for the Comprehensive Characterization of Terpenes and Terpenoids, Including Their Enantio-Distribution.

A microwave distillation method was optimized for the extraction and isolation of cannabis essential oil from fresh and dried hemp inflorescences. The developed method enabled us to obtain a distilled product rich in terpenes and terpenoid compounds, responsible of the typical and unique smell of the cannabis plant. The distillate from different hemp cultivars, including Kompolti, Futura 75, Carmagnola, Felina 32 and Finola were characterized by using a gas chromatograph equipped with both mass spectrometer and flame ionization detectors. In a single chromatographic run, the identity and absolute amounts of distilled compounds were determined. Peak assignment was established using a reliable approach based on the usage of two identification parameters, named reverse match, and linear retention index filter. Absolute quantification (mg g-1) of the analytes was performed using an internal standard method applying the flame ionization detector (FID) response factors according to each chemical family. An enantio-GC-MS method was also developed in order to evaluate the enantiomeric distribution of chiral compounds, an analytical approach commonly utilized for establishing the authenticity of suspicious samples.

Identification of high-value generating molecules from the wastes of tuna fishery industry by liquid chromatography and gas chromatography hyphenated techniques with automated sample preparation.

The present work aims to a promising re-utilization of the massive waste derived from the tuna fishing industry, for which by-products can represent more than 50% of the original material. Due to the considerable content in polyunsaturated fatty acids and noble proteins, such wastes can be used as primary source of functional ingredients in the production of nutraceuticals. The composition of the lipid and protein tuna fractions was investigated by means of gas chromatography-mass spectrometry and high-performance liquid chromatography-tandem mass spectrometry methods (in wastes and edible parts), and a preliminary characterization of potential bioactive peptides was achieved. Automated sample preparation allowed speeding up the analytical workflow, while allowing for highly sensitive and selective lipid characterization. The ω3 fatty acid content was found higher in waste products compared to the muscle, in terms of fatty acids as well as complex lipids. As for peptides, extraction by isoelectric solubilization/precipitation was performed, followed by enzymatic digestion and high-performance liquid chromatography-tandem mass spectrometry analysis. Furthermore, the use of bioinformatics tools highlighted the presence of potential antimicrobial peptides in the samples investigated.

Reliable identification and quantification of anabolic androgenic steroids in dietary supplements by using gas chromatography coupled to triple quadrupole mass spectrometry.

The aim of the present research was the identification and quantification of specific anabolic androgenic steroids (AASs) and other sterane structured compounds in dietary supplements (DSs). The adulteration of DSs by these compounds is of a particular concern in athletes, because it might lead to a positive doping result. The research was focused on the optimization of a highly sensitive and selective GC-based analytical strategy using triple quadrupole MS as detector. Chromatographic method and multiple reaction monitoring (MRM) transitions of 28 target compounds were optimized. Sample clean-up was carried out by using a solid phase extraction (SPE) procedure, while the derivatization of AASs was performed by using N-methyl-N-(trimethylsilyl)-trifluoroacetamide (MSTFA). The method was validated, and the following parameters were investigated: linearity range, limit of detection, accuracy, and precision expressed in terms of intra-day precision. The calibration curves were evaluated by using regression model and resulting in a good determination coefficients (R2 ≥ 0.9912). The residuals were scattered randomly around zero. The limits of detection (LODs) were lower than 7.0 ng g-1 or ng ml-1 . The accuracy assessment was evaluated in different forms of DSs characterized by high sample-to-sample variability (liquid, powder, tablet, capsule, protein, and herbal-based). Intra-day assay precision was in all cases lower than 20%. The developed analytical method was successfully applied to the analysis of 67 commercially available dietary supplements. In five cases, one or more steroid-type compounds were found in the concentration of 5 ng g-1 -100 µg g-1 , which might result adverse analytical findings in athletes.

Chemical Characterization of Three Accessions of Brassica juncea L. Extracts from Different Plant Tissues.

Indian mustard or Brassica juncea (B. juncea) is an oilseed plant used in many types of food (as mustard or IV range salad). It also has non-food uses (e.g., as green manure), and is a good model for phytoremediation of metals and pesticides. In recent years, it gained special attention due to its biological compounds and potential beneficial effects on human health. In this study, different tissues, namely leaves, stems, roots, and flowers of three accessions of B. juncea: ISCI 99 (Sample A), ISCI Top (Sample B), and "Broad-leaf" (Sample C) were analyzed by HPLC-PDA/ESI-MS/MS. Most polyphenols identified were bound to sugars and phenolic acids. Among the three cultivars, Sample A flowers turned were the richest ones, and the most abundant bioactive identified was represented by Isorhamnetin 3,7-diglucoside (683.62 µg/100 mg dry weight (DW) in Sample A, 433.65 µg/100 mg DW in Sample B, and 644.43 µg/100 mg DW in Sample C). In addition, the most complex samples, viz. leaves were analyzed by GC-FID/MS. The major volatile constituents of B. juncea L. leaves extract in the three cultivars were benzenepropanenitrile (34.94% in Sample B, 8.16% in Sample A, 6.24% in Sample C), followed by benzofuranone (8.54% in Sample A, 6.32% in Sample C, 3.64% in Sample B), and phytone (3.77% in Sample B, 2.85% in Sample A, 1.01% in Sample C). The overall evaluation of different tissues from three B. juncea accessions, through chemical analysis of the volatile and non-volatile compounds, can be advantageously taken into consideration for future use as dietary supplements and nutraceuticals in food matrices.

The opposite nitric oxide modulators do not lead to the opposite changes of metabolites under cadmium excess.

Responses of common medicinal herb Matricaria chamomilla to short-term cadmium (Cd) exposure (48 h) alone or in combination with nitric oxide (NO) scavenger (PTIO) or donor (SNP) were studied. Modulators revealed expected impact on NO formation (depletion under PTIO but elevation under SNP) and confirmed anomalous impact on Cd accumulation (stimulation by both modulators). Changes of proline and free amino acids in the roots and fatty acids (mainly α-linolenic acid) in the shoots indicate rather relation to elevated Cd uptake than to altered NO formation. On the contrary, root profile of fatty acids revealed the most prominent changes in response to PTIO (elevation of monounsaturated and polyunsaturated fatty acids) which confirm that NO depletion stimulates their biosynthesis. Soluble phenols were elevated by SNP co-application in the roots while accumulation of chlorogenic acid and umbelliferone revealed clear regulation by NO (i.e. enhancement in response to SNP and depletion in response to PTIO). Data indicate that quantities of some metabolites are affected by accumulated Cd or NO formation and that the opposite NO modulators do not automatically lead to the opposite changes of plant metabolites.

Antimicrobial Activity of Different Artemisia Essential Oil Formulations.

The extreme lipophilicity of essential oils (EOs) impedes the measurement of their biological actions in an aqueous environment. We formulated oil in water type Pickering Artemisia annua EO nanoemulsions (AEP) with surface-modified Stöber silica nanoparticles (20 nm) as the stabilizing agent. The antimicrobial activity of AEP and its effects on mature Candida biofilms were compared with those of Tween 80 stabilized emulsion (AET) and ethanolic solution (AEE) of the Artemisia EO. The antimicrobial activity was evaluated by using the minimum inhibitory concentrations (MIC90) and minimum effective concentrations (MEC10) of the compounds. On planktonic bacterial and fungal cells beside growth inhibition, colony formation (CFU/mL), metabolic activity, viability, intracellular ATP/total protein (ATP/TP), along with reactive oxygen species (ROS) were also studied. Artemisia annua EO nanoemulsion (AEP) showed significantly higher antimicrobial activity than AET and AEE. Artemisia annua EO nanoemulsions (AEP) generated superoxide anion and peroxides-related oxidative stress, which might be the underlying mode of action of the Artemisia EO. Unilamellar liposomes, as a cellular model, were used to examine the delivery efficacy of the EO of our tested formulations. We could demonstrate higher effectiveness of AEP in the EO components' donation compared to AET and AEE. Our data suggest the superiority of the AEP formulation against microbial infections.

Rapid and miniaturized qualitative and quantitative gas chromatography profiling of human blood total fatty acids.

The qualitative and quantitative profiling of fatty acids in human blood is a useful tool in disease prevention and health care, two concepts that are intimately related. In fact, fatty acid (FA) analysis can provide in-depth information on a specific metabolic state of individuals. The goal of the present research consisted of the development of a rapid and miniaturized analytical strategy for the complete characterization of the fatty acid profile in human blood. Sample collection was carried out by using the dried blood spot approach, while fatty acid derivatization to methyl esters was performed directly by using sodium methoxide and boron trifluoride. The following figures of merit were defined: intra- and inter-day repeatability, linearity range, limits of detection, and quantification. Additionally, the accuracy of the developed method was evaluated in the analysis of a certified reference human plasma sample. Apart from blood, the analytical procedure was also applied to samples of human serum and plasma. During the final stage of the research, the developed analytical method was performed in a fully automated manner. Graphical abstract.

Characterization of monoacylglycerols and diacylglycerols rich in polyunsaturated fatty acids produced by hydrolysis of Musteleus mustelus liver oil catalyzed by an immobilized bacterial lipase.

The use of an immobilized Serratia. sp W3 lipase as a replacement for the standard pancreatic lipases in the hydrolysis of liver oil from the Musteleus mustelus was studied. Monoacylglycerols (MAGs) and diacylglycerols (DAGs) containing ω-3 polyunsaturated fatty acids, namely eicosapentaenoic and docosahexaenoic acids were produced in hexane solvent at reaction temperatures reaching 55 °C with a molar triacylglycerol conversion over than 75 ± 5% in 24 h showing excellent hydrolysis characteristics. The favorable conditions for the hydrolysis reaction allowed fats with higher melting points to be analyzed facilitating the coupling of the hydrolysis reaction to the later steps in the analytical protocol. The lipid composition was elucidated for the first time by employing a highly efficient UHPLC-MS method with a novel embedded linear retention index approach. MAGs and DAGs obtained during the enzymatic hydrolysis could be used for the production of glycerol based emulsifiers of nutritional interest.

Chemical Composition of the Essential Oil of the Endemic Species Micromeria frivaldszkyana (Degen) Velen.

Micromeria frivaldszkyana is an endemic species found only in Bulgaria. Its essential oil (EO) composition is unknown. This study assessed the EO yield and composition of M. frivaldszkyana as a function of the location and of drying prior to the EO extraction. M. frivaldszkyana was sampled from two natural habitats, Uzana and Shipka in the Balkan Mountains; the EO was extracted via hydrodistillation and analyzed on GC/MS. The plants from the two locations had distinct EO composition. The EO content (in dried material) was 0.18% (Uzana) and 0.26% (Shipka). Monoterpene ketones were the major group of the EO constituents. Also, hydrocarbons predominated in the EO from Shipka, and alcohols predominated in the EO from Uzana. The EO from Uzana had a greater concentration of menthone (56% vs. 17% from Shipka) and neomenthol (7.8% vs. 2.4%). Conversely, the EO from Shipka had greater concentrations of pulegone (50% vs. 20% from Uzana), limonene (10.1% vs. 2.6%), and germacrene D (3.4% vs. 1.1%). Drying prior to the EO extraction altered the concentration of some constituents. This is the first report of M. frivaldszkyana EO yield and composition. The EO showed some similarities with the chemical profile of other Micromeria species, but overall, it has an unique chemical profile and may have distinctive applications.

Nitric oxide affects cadmium-induced changes in the lichen Ramalina farinacea.

Metabolic responses of epiphytic lichen Ramalina farinacea to cadmium (Cd) and/or nitric oxide (NO) scavenger (cPTIO) were studied. Accumulation of Cd and other metallic nutrients was not affected by cPTIO while total and absorbed amounts differed. Cd-induced NO formation was suppressed by cPTIO but ROS signal was synergistically enhanced, confirming that NO is essential to keep ROS under control. This excessive ROS generation could be a reason for depleted amount of all fatty acids, including SFAs, MUFAs and PUFAs. Total content of fatty acids reached 3.89 mg/g DW in control with linoleic (40%), palmitic (24%), oleic (12.8%) and stearic (8%) acids as major compounds: interestingly, shift in relative ratio of saturated (from 40 to 35% of total FAs) versus polyunsaturated fatty acids (from 42 to 48% of total FAs) was observed. Glutathione was suppressed by all treatments but Krebs acids were almost unaffected by cPTIO, indicating no regulatory role of NO in their accumulation. On the contrary, Cd-induced elevation in NO signal was related to increase in ascorbate and proline content while cPTIO suppressed it, indicating a tight relation between NO and these metabolites. Data are compared also with algae and vascular plants to show similarities between various life lineages.

Monoacylglycerol and diacylglycerol production by hydrolysis of refined vegetable oil by-products using an immobilized lipase from Serratia sp. W3.

Unlike the synthetic surfactants, mono- and diacylglycerols have the advantage to be biodegradable and non-toxic. In the present work, the hydrolysis of lipid fraction by-products of refined vegetable oils was performed by Serratia sp. W3 lipase immobilized on CaCO3 by combined adsorption and precipitation. This support was selected out of four carriers as it exhibited the finest activity support (950 U/g) and the most satisfactory behavior at use. The immobilized preparation with CaCO3 was stable and active in the whole range of pH (4 to 9) and temperature (37 to 55°C), yielding a 75% degree of hydrolysis at optimal environmental conditions of pH 8.5 and temperature 55°C. Thin-layer chromatography, gas chromatography, and liquid chromatography methods were evaluated to determine the analytical characterization of hydrolysis products. For monoacylglycerols and diacylglycerol fractions identified in the samples, a novel approach by liquid chromatography method was employed, through a homemade linear retention index database and a dedicated software. The adopted approach allowed the use of basic instrumentation set-ups, without the need of sophisticated detectors, such as mass spectrometers. Thus, it could be an effective alternative to produce emulsifiers from cheap vegetable oils.