Terpene Content Research Articles

Brain targeting of zolmitriptan via transdermal terpesomes: statistical optimization and in vivo biodistribution study by 99mTc radiolabeling technique

Zolmitriptan (ZT) is a potent second generation triptan, commonly administered to alleviate migraine attacks. ZT suffers various limitations; massive hepatic first pass metabolism, P-gp efflux transporters susceptibility, and limited (≈40%) oral bioavailability. Transdermal route of administration could be explored to enhance its bioavailability. A 23.31 full factorial design was constructed to developed twenty-four ZT loaded terpesomes via thin film hydration technique. The influence of drug: phosphatidylcholine ratio, terpene type, terpene concentration and sodium deoxycholate concentration on the characterization of the developed ZT-loaded terpesomes was assessed. Particle size (PS), zeta potential (ZP), ZT entrapment efficiency (EE%), drug loading (DL%) and drug released percentages after 6 h (Q6h) were the selected dependent variables. Further morphological, crystallinity, and in-vivo histopathological studies were conducted for the optimum terpesomes (T6). 99mTc-ZT and 99mTc-ZT-T6 gel were radio-formulated for in-vivo biodistribution studies in mice following transdermal application of 99mTc-ZT-T6 gel, relative to 99mTc-ZT oral solution. T6 terpesomes [comprising ZT and phosphatidylcholine (1:15), cineole (1% w/v) and sodium deoxycholate (0.1% w/v)] were optimum with respect to spherical PS (290.2 nm), ZP (-48.9 mV), EE% (83%), DL% (3.9%) and Q6h (92.2%) with desirability value of 0.85. The safety of the developed T6 terpesomes was verified by the in-vivo histopathological studies. 99mTc-ZT-T6 gel showed maximum brain concentration (5 ± 0.1%ID/ g) with highest brain to blood ratio of 1.92 ± 0.1 at 4 h post transdermal application. Significant improvement of ZT brain relative bioavailability (529%) and high brain targeting efficiency (315%) were revealed with 99mTc-ZT-T6 gel, which confirmed successful ZT delivery to the brain. Terpesomes could be safe, successful systems capable of improving ZT bioavailability with high brain targeting efficiency.Graphical

Impact of Wood Age on Termite Microbial Assemblages.

The decomposition of wood and detritus is challenging to most macroscopic organisms due to the recalcitrant nature of lignocellulose. Moreover, woody plants often protect themselves by synthesizing toxic or nocent compounds which infuse their tissues. Termites are essential wood decomposers in warmer terrestrial ecosystems and, as such, they have to cope with high concentrations of plant toxins in wood. In this paper, we evaluated the influence of wood age on the gut microbial (bacterial and fungal) communities associated with the termites Reticulitermes flavipes (Rhinotermitidae) (Kollar, 1837) and Microcerotermes biroi (Termitidae) (Desneux, 1905). We confirmed that the secondary metabolite concentration decreased with wood age. We identified a core microbial consortium maintained in the gut of R. flavipes and M. biroi and found that its diversity and composition were not altered by the wood age. Therefore, the concentration of secondary metabolites had no effect on the termite gut microbiome. We also found that both termite feeding activities and wood age affect the wood microbiome. Whether the increasing relative abundance of microbes with termite activities is beneficial to the termites is unknown and remains to be investigated. IMPORTANCE Termites can feed on wood thanks to their association with their gut microbes. However, the current understanding of termites as holobiont is limited. To our knowledge, no studies comprehensively reveal the influence of wood age on the termite-associated microbial assemblage. The wood of many tree species contains high concentrations of plant toxins that can vary with their age and may influence microbes. Here, we studied the impact of Norway spruce wood of varying ages and terpene concentrations on the microbial communities associated with the termites Reticulitermes flavipes (Rhinotermitidae) and Microcerotermes biroi (Termitidae). We performed a bacterial 16S rRNA and fungal ITS2 metabarcoding study to reveal the microbial communities associated with R. flavipes and M. biroi and their impact on shaping the wood microbiome. We noted that a stable core microbiome in the termites was unaltered by the feeding substrate, while termite activities influenced the wood microbiome, suggesting that plant secondary metabolites have negligible effects on the termite gut microbiome. Hence, our study shed new insights into the termite-associated microbial assemblage under the influence of varying amounts of terpene content in wood and provides a groundwork for future investigations for developing symbiont-mediated termite control measures.

Potential role of exogenous melatonin involved in postharvest quality maintenance of Vitis labrusca × vinifera 'Kyoho'.

Melatonin is an endogenous free radical scavenger with antioxidant activity that preserves the commercial value of postharvest fruits and delays fruit senescence. To explore the effect of exogenous melatonin on antioxidants and aroma volatile compounds of grapes (Vitis labrusca × vinifera 'Kyoho'), the grapes were treated with distilled water (control), 50 μmol L-1 of melatonin (M50), and 100 μmol L-1 of melatonin (M100) for 30 min and were then stored at 4 °C for 25 days. Exogenous melatonin decreased the rachis browning index, the decay development, the weight loss rate, the berry abscission rate, and the respiration rate, promoted the accumulation of total phenolics and total flavonoids, and delayed the reduction of anthocyanins and total soluble solids. In volatile compounds, the accumulation of esters, aldehydes, and alcohols in grapes was promoted, and the terpenes content was reduced by exogenous melatonin. Exogenous melatonin had potentially positive effects on the postharvest life and quality maintenance of grapes. These findings provide theoretical support for the application of melatonin in grape storage and preservation. © 2023 Society of Chemical Industry.

Effects of processing methods on the aroma constituents of hihatsumodoki (Piper retrofractum Vahl).

Hihatsumodoki (Piper retrofractum Vahl) is a traditional spice from Okinawa (Japan) that can be processed in different ways to create the desired flavor. Herein, we examined the effects of processing (sun-drying, oven-drying, roasting, and steaming) on the volatile aroma constituents of hihatsumodoki fruits. Among the 106 chromatographic peaks observed in total, 58 were assigned to known aroma compounds. The relative contents of terpenes, for example, linalool, β-caryophyllene, α-caryophyllene, and germacrene D, ranged from 57.6% to 88.1%. Sun-drying decreased the content of aldehydes such as hexanal and trans-2-hexenal but did not significantly affect the total content of aroma compounds. The amount of aroma compounds released during oven-drying and roasting increased with temperature up to a certain point (90°C) and decreased at an excessively high temperature of 180°C. High-temperature roasting generated Maillard reaction products such as furans and furanones, which could impart sweet caramel odors. Steamed fruits had the lowest content of aroma compounds, which was ascribed to the loss of these compounds to vapor. Meanwhile, drying steamed fruits resulted in an approximately 3.6-fold increase in their aroma compound content, and the content of sesquiterpenes in the steamed-dried fruits was similar to that in fruits exposed to high temperatures. The effects of processing on aroma quality were visualized using multivariate statistical analysis. The aroma characteristics of roasted (180°C), steamed, and steamed-dried fruits were different from those of the control. The combined findings provide useful information for the selection of processing methods to achieve the desired flavor of hihatsumodoki. Practical Application: This study reveals the effects of different processing methods on the aroma profile of hihatsumodoki (Piper retrofractum Vahl), a subtropical spice from Okinawa (Japan). The results facilitate the selection of preferred hihatsumodoki flavors for household and industrial applications in foods and beverages. In addition, they inspire research on the processing-induced flavor changes of other tropical or subtropical spices.

6 P-Cymene Formation by Oxidation of Cyclohexadiene Monoterpenes – a Pitfall in Air Sampling of Terpenes

Abstract It has been known for a long time that certain terpenes are sensitive to oxidation. Especially in the presence of small amounts, as is the case in the environment or indoors, changes occur due to UV light or ozone exposition. Various oxidized products have also been detected when sampling low terpene concentrations using Tenax® tubes and subsequent thermal desorption. Here we show that carrying out common air sampling of monoterpenes in the lower ppm range on charcoal tubes, the atmospheric oxygen is sufficient to oxidize the most sensitive terpenes to a considerable extent, forming almost exclusively p-cymene. As expected, the extent of aromatization was strongest in the terpenes with a double unsaturated ring system such as α- or γ-terpinene or α-phellandrene while p-menthene or limonene were not affected. We studied the influence of sampling time, temperature, humidity and storage time of the coated tubes. Simultaneously conducted experiments with thermal desorption in the ppb range showed a significantly reduced p-cymene formation compared to the activated charcoal results. Our findings were confirmed by air samples of terpenes taken during the grinding of nutmegs. The concentrations of p-cymene determined were significantly higher than the terpene pattern of nutmeg oil would have suggested. Thus, the detection of p-cymene during air sampling may be an artifact caused by the presence of cyclohexadiene monoterpenes. This is particularly important because p-cymene is often used as an internal standard in terpene analysis thus preventing its artifact formation from being noticed.

Freeze-drying Cannabis sativa L. using real-time relative humidity monitoring and mathematical modeling for the cannabis industry

Pre-freezing has an impact on the drying time and physiochemical properties of cannabis. This study investigated the relationship between sample mass reduction and relative humidity during freeze-drying and the effects of pre-freezing and freeze-drying temperature on cannabis drying kinetics, trichome structure, and color, in addition to cannabinoid and terpene concentrations. Three cannabis accessions, Qrazy Train, Qrazy Apple, and Qrazy Angel, were dried at 10°C, and 20°C, with different pre-freezing conditions (−20°C and −40°C). Pre-freezing rates of 0.13°C min−1 for both Qrazy Train and Qrazy Angel and 0.15°C min−1 for Qrazy Apple were recorded for pre-freezing at − 20°C and significantly (p < 0.05) increased by 73.5%, 71.2%, and 72.9% for Qrazy Train, Qrazy Apple, and Qrazy Angel, respectively, when inflorescence was pre-frozen at − 40°C. Rational regression model best explains the relationship between mass reduction and relative humidity during drying, while drying kinetics can be described using the Page and Logarithmic models. Total color changes ranged from 1.16 to 9.69 total color changes measured for all dried samples compared to respective fresh, undried samples were not significantly (p < 0.05) different. THCA concentrations for fresh, undried Cannabis sativa L. accessions ranged from 214.4 mg g−1 to 257.5 mg g−1; this was higher than their CBDA concentrations, ranging from 0.03 mg g−1 to 0.1 mg g−1. Freeze-drying increased [CBDA], [CBGA], and [CBG] in dried samples ranging from 0.45 mg g−1 to 0.38 mg g−1, 2.87 mg g−1 to 4.91 mg g−1, and 0.57 mg g−1 to 1.33 mg g−1, respectively. Mean terpene concentration ranged from 17.7 mg g−1 to 40.3 mg g−1. Irrespective of the pre-freezing condition or cannabis accession, drying at 20°C reduced drying time by 10.4–31.9%. Findings could be of industrial relevance for improving post-harvest processes while maintaining quality of this regulated crop.

Evaluation of volatile compound profiles and sensory properties of dark and pale beers fermented by different strains of brewing yeast

To evaluate the differences in the volatile compound profile of dark and pale beers fermented by different strains of brewer's yeast, gas chromatography with flame ionization detection and gas chromatography mass spectrometry analysis of eight beers was carried out. The prevalent group of compounds in all the beers analysed were alcohols (56.41–72.17%), followed by esters (14.58–20.82%), aldehydes (8.35–20.52%), terpenes and terpenoids (1.22–6.57%) and ketones (0.42–1.00%). The dominant higher alcohols were 2-methylpropan-1-ol, 3-methylbutanol, phenethyl alcohol, among aldehydes furfural, decanal, nonanal, and among esters ethyl acetate, phenylethyl acetate and isoamyl acetate. Beers fermented by the top-fermenting yeast Saccharomyces cerevisiae var. diastaticus had the highest volatile content. The addition of dark malt in wort production process had no effect on the total content of volatiles, but for some beers it caused changes in the total content of esters, terpenes and terpenoids. Variations in the total volatile content between beers fermented by different yeast strains are mainly due to esters and alcohols identified. Sensory analysis of beers allowed us to identify the characteristics affected by the addition of dark speciality malts in the production of wort and yeast strains used in the fermentation process.

Selected cannabis terpenes synergize with THC to produce increased CB1 receptor activation

The cannabis plant exerts its pharmaceutical activity primarily by the binding of cannabinoids to two G protein-coupled cannabinoid receptors, CB1 and CB2. The role that cannabis terpenes play in this activation has been considered and debated repeatedly, based on only limited experimental results. In the current study we used a controlled in-vitro heterologous expression system to quantify the activation of CB1 receptors by sixteen cannabis terpenes individually, by tetrahydrocannabinol (THC) alone and by THC-terpenes mixtures. The results demonstrate that all terpenes, when tested individually, activate CB1 receptors, at about 10–50% of the activation by THC alone. The combination of some of these terpenes with THC significantly increases the activity of the CB1 receptor, compared to THC alone. In some cases, several fold. Importantly, this amplification is evident at terpene to THC ratios similar to those in the cannabis plant, which reflect very low terpene concentrations. For some terpenes, the activation obtained by THC- terpene mixtures is notably greater than the sum of the activations by the individual components, suggesting a synergistic effect. Our results strongly support a modulatory effect of some of the terpenes on the interaction between THC and the CB1 receptor. As the most effective terpenes are not necessarily the most abundant ones in the cannabis plant, reaching “whole plant” or “full spectrum” composition is not necessarily an advantage. For enhanced therapeutic effects, desired compositions are attainable by enriching extracts with selected terpenes. These compositions adjust the treatment for various desired medicinal and personal needs.

Creation of New Oregano Genotypes with Different Terpene Chemotypes via Inter- and Intraspecific Hybridization.

Oregano is a medicinal and aromatic plant of value in the pharmaceutical, food, feed additive, and cosmetic industries. Oregano breeding is still in its infancy compared with traditional crops. In this study, we evaluated the phenotypes of 12 oregano genotypes and generated F1 progenies by hybridization. The density of leaf glandular secretory trichomes and the essential oil yield in the 12 oregano genotypes varied from 97-1017 per cm2 and 0.17-1.67%, respectively. These genotypes were divided into four terpene chemotypes: carvacrol-, thymol-, germacrene D/β-caryophyllene-, and linalool/β-ocimene-type. Based on phenotypic data and considering terpene chemotypes as the main breeding goal, six oregano hybrid combinations were performed. Simple sequence repeat (SSR) markers were developed based on unpublished whole-genome sequencing data of Origanum vulgare, and 64 codominant SSR primers were screened on the parents of the six oregano combinations. These codominant primers were used to determine the authenticity of 40 F1 lines, and 37 true hybrids were identified. These 37 F1 lines were divided into six terpene chemotypes: sabinene-, β-ocimene-, γ-terpinene-, thymol-, carvacrol-, and p-cymene-type, four of which (sabinene-, β-ocimene-, γ-terpinene-, and p-cymene-type) were novel (i.e., different from the chemotypes of parents). The terpene contents of 18 of the 37 F1 lines were higher than those of their parents. The above results lay a strong foundation for the creating of new germplasm resources, constructing of genetic linkage map, and mapping quantitative trait loci (QTLs) of key horticultural traits, and provide insights into the mechanism of terpenoid biosynthesis in oregano.

Selection of Saccharomyces cerevisiae Isolates from Helan Mountain in China for Wine Production

S. cerevisiae strains were isolated and identified from vineyards and the spontaneous fermentation must at the eastern foot of Helan Mountain in China, and their oenological properties and fermentation abilities were analyzed. From the total of 199 S. cerevisiae strains isolated and identified, 14 isolates (F4-13, F5-7, F5-9, F5-12, F5-18, F5-19, F5-21, F6-8, F6-23, F9-23, SXY-4, HT-10, ZXY-17, MXY-19) exhibited excellent tolerance to sugar, SO2, and ethanol. Among the isolates, the strain F4-13 exhibited the better oenological properties, with low H2S production (+), suitable flocculation ability (58.74%), and reducing-L-malic acid ability (49.07%), and generated high contents of polyphenol, anthocyanin, tannin, terpenes, and higher alcohols, which contributed to the improvement of the red fruity and floral traits of the wines. The obtained results provide a strategy for the selection of indigenous S. cerevisiae for wine fermentation to produce high-quality wine with regional characteristics.

Novel Low-Alcohol Sangria-Type Wine Products with Immobilized Kefir Cultures and Essential Oils

Low-alcohol wines (ranging from &lt;0.5 to 10.5% vol) are novel products that have been steadily gaining scientific and commercial attention. Over the past few years, consumer interest in healthier foods has augmented the development of novel functional products containing probiotic microorganisms, while the urge for a reduction in chemical preservatives has shifted the food and wine industry’s interest to natural alternatives, such as essential oils (EOs). In the present study, low-alcohol (~6% vol) wines with (wet or dried) immobilized kefir cultures on fruit pieces, and essential oils (Citrus medica or Cinnamomum zeylanicum) were produced and evaluated for their properties. The viability of the immobilized kefir cultures on apple and pear pieces was not affected by the addition of EOs, and levels &gt;7 logcfu/g were maintained after 2 h of immersion in wines. HS-SPME GC/MS analysis revealed characteristic compounds originating from the chemical composition of the added EOs in the final products. Principal component analysis (PCA) revealed that the relative content of terpenes, alcohols and carbonyl compounds played a major role in the discrimination of low-alcohol wine products. EO addition affected the products’ sensory evaluation and resulted in significantly higher aroma and taste density compared to control samples. Notably, all novel Sangria-type wine variants were approved during preliminary sensory evaluation.

THE PRODUCTION OF LOW-ALCOHOL WINES BY SEQUENTIAL SEMI-ANAEROBIC MUST FERMENTATION

Nowadays, consumers go towards products that provide food security and nutritional richness, consumption of highly alcoholic beverages not complying with these health-related requirements. This study aimed to obtain low-alcohol wines by performing sequential alcoholic fermentation of grape must (Muscat Ottonel). Thus, 11 commercial yeast strains (10 Saccharomyces cerevisiae and 1 Torulaspora delbrueckii) were preliminarily tested in anaerobic and semi-anaerobic conditions. Based on laboratory tests, grape must fermentation was sequentially initiated in semi-anaerobic conditions (loose cotton plugs; intermittent homogenization), with the T. delbrueckii strain (106 CFU/mL) and the addition of preliminarily selected S. cerevisiae strains (104 CFU/mL), to the consumption of 50% of the sugars in the must, to complete the fermentation. By applying this technology, dry wines were obtained with an alcohol content lower by up to 1.10% vol., but with a lower concentration of volatile terpenes, due to additional must oxidation. Also, semi-anaerobiosis enhanced glycerol synthesis by yeasts (&lt; 35%), with a positive impact on the sensory characteristics of the wine.

Physico-Chemical Characterization of an Exocellular Sugars Tolerant Β-Glucosidase from Grape Metschnikowia pulcherrima Isolates.

A broad variety of microorganisms with useful characteristics in the field of biotechnology live on the surface of grapes; one of these microorganisms is Metschnikowia pulcherrima. This yeast secretes a β-glucosidase that can be used in fermentative processes to liberate aromatic compounds. In this work, the synthesis of an exocellular β-glucosidase has been demonstrated and the optimal conditions to maximize the enzyme's effectiveness were determined. There was a maximum enzymatic activity at 28 °C and pH 4.5. Furthermore, the enzyme presents a great glucose and fructose tolerance, and to a lesser extent, ethanol tolerance. In addition, its activity was stimulated by calcium ions and low concentrations of ethanol and methanol. The impact of terpene content in wine was also determined. Because of these characteristics, β-glucosidase is a good candidate for use in enology.

Observations of biogenic volatile organic compounds over a mixed temperate forest during the summer to autumn transition

Abstract. The exchange of trace gases between the biosphere and the atmosphere is an important process that controls both chemical and physical properties of the atmosphere with implications for air quality and climate change. The terrestrial biosphere is a major source of reactive biogenic volatile organic compounds (BVOCs) that govern atmospheric concentrations of the hydroxy radical (OH) and ozone (O3) and control the formation and growth of secondary organic aerosol (SOA). Common simulations of BVOC surface–atmosphere exchange in chemical transport models use parameterizations derived from the growing season and do not consider potential changes in emissions during seasonal transitions. Here, we use observations of BVOCs over a mixed temperate forest in northern Wisconsin during broadleaf senescence to better understand the effects of the seasonal changes in canopy conditions (e.g., temperature, sunlight, leaf area, and leaf stage) on net BVOC exchange. The BVOCs investigated here include the terpenoids isoprene (C5H8), monoterpenes (MTs; C10H16), a monoterpene oxide (C10H16O), and sesquiterpenes (SQTs; C15H24), as well as a subset of other monoterpene oxides and dimethyl sulfide (DMS). During this period, MTs were primarily composed of α-pinene, β-pinene, and camphene, with α-pinene and camphene dominant during the first half of September and β-pinene thereafter. We observed enhanced MT and monoterpene oxide emissions following the onset of leaf senescence and suggest that senescence has the potential to be a significant control on late-season MT emissions in this ecosystem. We show that common parameterizations of BVOC emissions cannot reproduce the fluxes of MT, C10H16O, and SQT during the onset and continuation of senescence but can correctly simulate isoprene flux. We also describe the impact of the MT emission enhancement on the potential to form highly oxygenated organic molecules (HOMs). The calculated production rates of HOMs and H2SO4, constrained by terpene and DMS concentrations, suggest that biogenic aerosol formation and growth in this region should be dominated by secondary organics rather than sulfate. Further, we show that models using parameterized MT emissions likely underestimate HOM production, and thus aerosol growth and formation, during early autumn in this region. Further measurements of forest–atmosphere BVOC exchange during seasonal transitions as well as measurements of DMS in temperate regions are needed to effectively predict the effects of canopy changes on reactive carbon cycling and aerosol production.

Identification of QTL controlling volatile terpene contents in tea plant (Camellia sinensis) using a high-aroma 'Huangdan' x 'Jinxuan' F1 population.

Aroma is an important factor affecting the character and quality of tea. The improvement of aroma trait is a crucial research direction of tea plant breeding. Volatile terpenes, as the major contributors to the floral odors of tea products, also play critical roles in the defense responses of plants to multiple stresses. However, previous studies have largely focused on the aroma formation during the manufacture of tea or the comparison of raw tea samples. The mechanisms causing different aroma profiles between tea cultivars have remained underexplored. In the current study, a high-density genetic linkage map of tea plant was constructed based on an F1 population of 'Huangdan' × 'Jinxuan' using genotyping by sequencing. This linkage map covered 1754.57 cM and contained 15 linkage groups with a low inter-marker distance of 0.47 cM. A total of 42 QTLs associated with eight monoterpene contents and 12 QTLs associated with four sesquiterpenes contents were identified with the average PVE of 12.6% and 11.7% respectively. Furthermore, six candidate genes related to volatile terpene contents were found in QTL cluster on chromosome 5 by RNA-seq analysis. This work will enrich our understanding of the molecular mechanism of volatile terpene biosynthesis and provide a theoretical basis for tea plant breeding programs for aroma quality improvement.

Production and characterization of a novel cold-active ß-glucosidase and its influence on aromatic precursors of Muscat wine

ß-Glucosidases (ßGL)—widely used in the enological field—are enzymes that catalyze the liberation of aromatic volatiles from their glycosidic precursors during winemaking. In the present study, a ßGL obtained from the Antarctic yeast Mrakia sp. LP 7.1.2016 was produced on a bioreactor scale, purified, characterized, and the enzyme's properties studied for a potential enological application. Sodium-dodecylsulfate–polyacrylamide-gel electrophoresis indicated a high molecular weight for this enzyme, it having at least two subunits of 134 and 14 kDa. ßGL exhibited a pH optimum of 5.0 and retained substantial activity and stability at pH 4.0. The temperature range for optimal activity was 50–55 °C, with the enzyme demonstrating thermostability up to 50 °C and retaining 87% of residual activity at that temperature after 3 h. The respective kinetic constants determined with p-nitrophenyl-ß-d-glucopyranoside and cellobiose were 0.38 and 1.79 mM for Km and 20.1 and 5.65 μmol−1 mg−1 min.1 for Vmax, ß-Glucosidase manifested high activity in 10–25% (v/v) ethanol, 30.0 mg L−1 sulfur dioxide, and 10–200 g L−1 fructose; but it was strongly inhibited by glucose, retaining only 6% of residual activity in the presence of 20 g L−1. Upon investigating the influence of the enzyme on Muscat-wine glycosidic precursors, we found significant differences in terpene content after 14 days of ßGL treatment at an eightfold increase over control-wine levels. The enzyme was more active toward the precursors of the monoterpenes nerol and geraniol and oxides of trans- and cis-linalool. These findings contribute to our understanding of the potential of cold-active enzymes in advantageous biotechnological applications to enology.

Naïve Pine Terpene Response to the Mountain Pine Beetle (Dendroctonus ponderosae) through the Seasons.

Insect herbivores must contend with constitutive and induced plant defenses. The mountain pine beetle, Dendroctonus ponderosae Hopkins (Coleoptera: Curculionidae, Scolytinae) has expanded its range east of the Rocky Mountains into the western boreal forest and is encountering evolutionarily naïve lodgepole pines (Pinus contorta) and jack pines (Pinus banksiana). Pinus contorta and P. banksiana in the expanded range have different constitutive and induced defenses in response to wounding and inoculation with fungal associates of D. ponderosae. In the historic range, previous studies have examined phloem terpene content prior to and just after D. ponderosae mass attack, but the terpene profile of attacked trees post-overwintering is unknown. We examined the response of mature P. contorta and P. banksiana trees to experimentally-applied mass attack by D. ponderosae and quantified phloem terpenes at three time points, pre-attack, post-attack (same season), and the following spring, post-overwintering. Phloem content of total terpenes as well as many individual terpenes increased after D. ponderosae attack but were only significantly higher than pre-attack levels at the post-overwintering time point in both P. contorta and P. banksiana. The absence of a significant increase in phloem terpenes in the month following attack in naïve pines is a potential cause for increased D. ponderosae offspring production reported in naïve P. contorta. Beetle attack density did not influence the phloem terpene profiles of either species and there was no significant interaction between attack density and sampling time on terpene content. High phloem terpenes in trees that are attacked at low densities could prime these trees for defense against attacks in the following season but it could also make these trees more apparent to early-foraging beetles and facilitate efficient mass attack at low D. ponderosae population densities in the expanded range.

Relationship between Total Antioxidant Capacity, Cannabinoids and Terpenoids in Hops and Cannabis.

Efficient determination of antioxidant activity in medicinal plants may provide added value to extracts. The effects of postharvest pre-freezing and drying [microwave-assisted hot air (MAHD) and freeze drying] on hops and cannabis were evaluated to determine the relationship between antioxidant activity and secondary metabolites. The 2,2-diphenyl-1-picrylhydrazine (DPPH) reduction and ferric reducing ability of power (FRAP) assays were assessed for suitability in estimating the antioxidant activity of extracted hops and cannabis inflorescences and correlation with cannabinoid and terpene content. Antioxidant activity in extracts obtained from fresh, undried samples amounted to 3.6 Trolox equivalent antioxidant activity (TEAC) (M) dry matter-1 and 2.32 FRAP (M) dry matter-1 for hops, in addition to 2.29 TEAC (M) dry matter-1 and 0.25 FRAP (M) dry matter-1 for cannabis. Pre-freezing significantly increased antioxidant values by 13% (DPPH) and 29.9% (FRAP) for hops, and by 7.7% (DPPH) and 19.4% (FRAP) for cannabis. ANOVA analyses showed a significant (p < 0.05) increase in total THC (24.2) and THCA (27.2) concentrations (g 100 g dry matter-1) in pre-frozen, undried samples compared to fresh, undried samples. Freeze-drying and MAHD significantly (p < 0.05) reduced antioxidant activity in hops by 79% and 80.2% [DPPH], respectively and 70.1% and 70.4% [FRAP], respectively, when compared to antioxidant activity in extracts obtained from pre-frozen, undried hops. DPPH assay showed that both freeze-drying and MAHD significantly (p < 0.05) reduced the antioxidant activity of cannabis by 60.5% compared to the pre-frozen samples although, there was no significant (p < 0.05) reduction in the antioxidant activity using the FRAP method. Greater THC content was measured in MAHD-samples when compared to fresh, undried (64.7%) and pre-frozen, undried (57%), likely because of decarboxylation. Both drying systems showed a significant loss in total terpene concentration, yet freeze-drying has a higher metabolite retention compared to MAHD. These results may prove useful for future experiments investigating antioxidant activity and added value to cannabis and hops.

Digital Technologies Applied to Control the One-Step Process of Cannabis Olive Oil Preparations

The reproducibility of an extemporaneous preparation is an essential condition for guaranteeing the quality, efficacy, and safety of the medicinal product. This study aimed to develop a controlled one-step process for cannabis olive oil preparations by applying digital technologies. For this purpose, the chemical profile of cannabinoid contents in oil extracts of Bedrocan, FM2, and Pedanios varieties obtained with the already in use method, proposed by the Italian Society of Compounding Pharmacists (SIFAP), was compared with two new methods, specifically the Tolotto Gear® extraction method (TGE) and the Tolotto Gear® extraction method preceded by a pre-extraction procedure (TGE-PE). HPLC analyses showed that the concentration of THC using cannabis flos with a high THC content (over 20% w/w) was always higher than 21 mg/mL for the Bedrocan variety and close to 20 mg/mL for the Pedanios variety when applying TGE, while with TGE-PE, the THC concentration was higher than 23 mg/mL for the Bedrocan variety. For the FM2 variety, the amounts of THC and CBD in the oil formulations obtained using TGE were higher than 7 mg/mL and 10 mg/mL, respectively, and for TGE-PE, the concentrations of THC and CBD were higher than 7 mg/mL and 12 mg/mL, respectively. GC-MS analyses were performed to define the terpene contents in the oil extracts. The samples of Bedrocan flos extracted with TGE-PE displayed a distinctive profile, highly rich in terpenes and devoid of oxidized volatile products. Thus, TGE and TGE-PE allowed performing a quantitative extraction of cannabinoids and increasing the total mono-di-tri terpenes and sesquiterpene concentrations. The methods were repeatable and applicable to any quantity of raw material, preserving the phytocomplex of the plant.

Contribution of Starmerella bacillaris and Oak Chips to Trebbiano d'Abruzzo Wine Volatile and Sensory Diversity.

In this study, six fermentation trials were carried out: co-inoculation and sequential inoculation of Saccharomyces cerevisiae and Starmerella bacillaris in the presence and absence of oak chips. Moreover, Starm. bacillaris strain was attached to the oak chips and co-inoculated or sequentially inoculated with S. cerevisiae. Wines fermented with Starm. bacillaris adhered to oak chips showed a higher concentration of glycerol (more than 6 g/L) than the others (about 5 g/L). These wines also showed a higher content of polyphenols (more than 300 g/L) than the others (about 200 g/L). The addition of oak chips induced an increase of yellow color (b* value of about 3). Oak-treated wines were characterized by a higher concentration of higher alcohols, esters and terpenes. Aldehydes, phenols and lactones were detected only in these wines, independently from the inoculation strategy. Significant differences (p < 0.05) were also observed in the sensory profiles. The fruity, toasty, astringency, and vanilla sensations were perceived as more intense in wines treated with oak chips. The white flower descriptor showed a higher score in wines fermented without chips. Oak surface-adhered Starm. bacillaris cells could be a good strategy to improve the volatile and sensory profile of Trebbiano d'Abruzzo wines.