Terpenoid Profiles Research Articles

Revealing the aromatic sonata through terpenoid profiling and gene expression analysis of aromatic and non-aromatic coconut varieties

Aromatic coconut represents an exceptional variety of coconut known for its distinct and delightful flavor and aroma, both of which are highly cherished by consumers. Despite its popularity, there has been a lack of systematic research on aroma components and the associated synthetic genes. In this report, we developed the metabolite profiles of terpenoids by targeted metabolomics and obtained the expression profile of genes related to terpenoid biosynthesis by RNA-seq during different coconut fruit developmental stages. Totally, we separated 26 different terpenoids in aromatic coconut pulp, among which, geranyl acetate and (−)-isosyngene emerged as the most abundant. The integrated analysis of metabolism and RNA-seq data showed that HMGS2, HMGS3, IPI/IDI1, HMGR1, HMGR3, and CMK2 as potentially key genes involved in the synthesis of terpenoids in aromatic coconut. To validate these findings, qRT-PCR was conducted on terpenoid-related genes. These findings lay a foundation for understanding aroma formation and the molecular mechanism of terpenoids in coconut fruit.

Monoterpenoids isomerization and cyclization processes in Gewürztraminer wines: A kinetic investigation at different pH and temperatures

The evolution of volatile compounds in wine comprises several acid-catalyzed reactions, such as glycosides precursors’ hydrolysis and rearrangements, and significantly contributes to its sensory qualities, even after prolonged aging. The aim of this work was to use a well-defined experimental design and to examine how terpenoids in Gewürztraminer wine change over time when subjected to different temperatures and pH levels over two weeks. A theoretically-based approach was used, involving the definition of a complete system of ordinary differential equations (ODE) with well-established boundary conditions (initial concentration of reactants/products), using Kinetiscope, a kinetic simulation software. The calculated optimal rate constants, based on the experimental curves, produced a comprehensive data set describing the evolution of the terpenoid profile, highlighting the reversible and irreversible interconversion processes. Finding revealed that higher pH and lower temperature conditions are crucial for preserving the important terpenoid compounds that characterize the typicality and exemplarity of Gewürztraminer aroma profile.

Transcriptome Analyses Reveal Differences in the Metabolic Pathways of the Essential Oil Principal Components of Different Cinnamomum Chemotypes

The genus Cinnamomum exhibits a rich variety of chemotypes and is an economically important essential oil (EO)-producing plant belonging to the family Lauraceae. Here, we aimed to explore the potential differences in the terpenoid (the principal components of EOs) biosynthesis pathways of different chemotypes at the molecular level in four Cinnamomum species—C. camphora var. linaloolifera, C. kanehirae, C. longipaniculatum, and C. micranthum. Gas chromatography–mass spectrometry (GC-MS) was employed to elucidate the discrepancies in the chemical profiles and compositions of leaf EO terpenoids among the four Cinnamomum species. The results revealed significant variations in leaf EO yields. The main constituents of the leaf EOs from C. camphora var. linaloolifera and C. kanehirae were the acyclic monoterpene linalool, and those of C. longipaniculatum and C. micranthum were the monoterpene eucalyptol and the sesquiterpene β-caryophyllene, respectively. Furthermore, a comparative transcriptome analysis of the leaves from the four Cinnamomum species revealed that differentially expressed genes (DEGs) were significantly enriched in terpene-related entries. Specifically, 42 and 24 DEGs were significantly enriched to the mevalonate (MVA)/2-methylerythritol 4-phosphate (MEP) pathways and terpene synthase (TPS) activity, respectively. Most genes encoding proteins involved in the terpenoid precursor MVA and MEP pathways exhibited differential expression across the four species, which correlated with the distinct terpenoid profiles observed in their leaf EOs. Four acyclic monoterpene linalool synthase genes—Maker00024100, Maker00014813, Maker00014818, and Maker00018424—were highly expressed in C. camphora var. linaloolifera and C. kanehirae. A monoterpene eucalyptol synthesis gene, Maker00001509, was highly expressed in C. longipaniculatum, and a sesquiterpene β-stigmasterol synthesis gene, Maker00005791, was highly expressed in C. micranthum. These expression levels were subsequently validated through quantitative real-time polymerase chain reaction (qRT-PCR). In conclusion, the combined results of the GC-MS and transcriptome analyses revealed a strong correlation between the metabolite content of the EOs and gene expression. This research contributes to a better understanding of the differences in terpene accumulation in various chemotypes of Cinnamomum at the molecular and mechanistic levels, laying a solid foundation for the cultivation of an ideal Cinnamomum variety.

Exogenous stimulation of Tanacetum vulgare roots with pipecolic acid leads to tissue-specific responses in terpenoid composition.

Tanacetum vulgare L., tansy, is a perennial plant with highly variable terpenoid composition, with mono- and sesquiterpenoids being the most abundant. The high diversity of terpenoids plays an important role in mediating ecological interactions. However, the distribution of terpenoids in different tissues and inducibility of terpenoids in these tissues via biotic stress are poorly understood. We investigated changes in terpenoid profiles and concentrations in different organs following treatment of roots with pipecolic acid (Pip), a non-proteinogenic amino acid that triggers defence responses leading to induce systemic resistance (SAR) in plants. Tansy leaves and midribs contained mainly monoterpenoids, while coarse and fine roots contained mainly sesquiterpenoids. Rhizomes contained terpenoid profiles of both midribs and roots but also unique compounds. Treatment with Pip led to an increase in concentrations of mono- and sesquiterpenoids in all tissues except rhizomes. However, significantly more sesquiterpenoids was formed in root tissues in response to Pip treatment, compared to shoots. The metabolic atlas for terpenoids presented here shows that there is exceptionally strong differentiation of terpenoid patterns and terpenoid content in different tissues of tansy. This, together with differential inducibility by Pip, suggests that the chemical diversity of terpenoids may play an important role in tansy ecological interactions and defence against biotic stressors that feed on below- and aboveground organs.

Intraspecific chemical variation of Tanacetum vulgare affects plant growth and reproductive traits in field plant communities.

The study investigated the impact of intraspecific plant chemodiversity on plant growth and reproductive traits at both the plant and plot levels. It also aimed to understand how chemodiversity at stand level affects ecosystem functioning and plant-plant interactions. We describe a biodiversity experiment in which we manipulated intraspecific plant chemodiversity at the plot level using six different chemotypes of common tansy (Tanacetum vulgare L., Asteraceae). We tested the effects of chemotype identity and plot-level chemotype richness on plant growth and reproductive traits and plot-level headspace emissions. The study found that plant chemotypes differed in growth and reproductive traits and that traits were affected by the chemotype richness of the plots. Although morphological differences among chemotypes became less pronounced over time, reproductive phenology patterns persisted. Plot-level trait means were also affected by the presence or absence of certain chemotypes in a plot, and the direction of the effect depended on the specific chemotype. However, chemotype richness did not lead to overyielding effects. Lastly, chemotype blends released from plant communities were neither richer nor more diverse with increasing plot-level chemotype richness, but became more dissimilar as they became more dissimilar in their leaf terpenoid profiles. We found that intraspecific plant chemodiversity is crucial in plant-plant interactions. We also found that the effects of chemodiversity on plant growth and reproductive traits were complex and varied depending on the chemotype richness of the plots. This long-term field experiment will allow further investigation into plant-insect interactions and insect community assembly in response to intraspecific chemodiversity.

Geographic distribution of terpenoid chemotypes in Tanacetum vulgare mediates tansy aphid occurrence but not abundance

Intraspecific variation of specialized metabolites in plants, such as terpenoids, are used to determine chemotypes. Tansy Tanacetum vulgare exhibits diverse terpenoid profiles that affect insect communities. However, it is not fully known whether patterns of their chemical composition and associated insects vary beyond the community scale. Here, we investigated the geographic distribution of mono‐ and sesquiterpenoid chemotypes in tansy leaves and their relationships with specific insect communities across Germany. We sampled tansy leaves from ten plants with and five plants without aphids in each of 26 sites along a north–south and west–east transect in Germany. Hexane‐extracted metabolites from leaf tissues were analyzed by gas chromatography‐mass spectrometry (GC‐MS). Plant morphological traits, aphid occurrence and abundance, and occurrence of ants were recorded locally. The effect of plant chemotype, plant morphological parameters, and abiotic site parameters such as soil types, temperature and precipitation on insect occurrences were analyzed. Plants clustered into four monoterpenoid and four sesquiterpenoid chemotype classes. Monoterpene classes differed in their latitudinal distribution, whereas sesquiterpenes were more evenly distributed across the transect. Aphid and ant occurrence was influenced by monoterpenoids. Plants of monoterpenoid class 1 were colonized by aphids and ants significantly more often than expected by chance, whereas in other classes there were no significant differences. Aphid abundance was affected by soil type, and average annual temperature positively correlated with the occurrence of ants. We found significant geographic patterns in the distribution of tansy chemodiversity and show that monoterpenoids affect aphid and ant occurrence, while the soil type can influence aphid abundance. We show that geographic variation in plant chemistry influences insect community assembly on tansy plants.

Sequential extraction approach for sustainable recovery of various hemp (Cannabis sativa L.) bioactive compounds

Cannabis sativa L. known as industrial hemp manifests high capability in the manufacturing of valuable nutraceuticals with health benefits. This study was focused on investigation of chemical profile and bioactive potential of extracts isolated from three hemp varieties (Helena, Marina and Fedora 17). In addition, great attention was directed towards the use of green extraction techniques and maximum utilization of raw material. In that regard conventional (hydrodistillation – HD) and novel extraction techniques (microwave-assisted hydrodistillation – MWHD, supercritical fluid extraction – SFE and microwave-assisted extraction – MAE) were applied for isolating various fractions (essential oil, lipid extracts and liquid extract) with promising bioactive potential. Essential oils and lipid extracts were characterized regarding yield, terpenoid profile determined by GC-MS and in vitro antioxidant activity towards DPPH and ABTS + radicals. Furthermore, the content of two major cannabinoids was determined in plant material and in extract obtained by SFE. Sequential SFE technique allows obtaining lipid extracts in the first step of process as well as raffinates after SFE in the second step. MAE was applied for isolation of polyphenolic fraction from raw materials and SFE raffinates. Polyphenolic content was determined by spectrophotometric assays and UHPLC-Triple-quadrupole-MS. The results suggested that hemp is an excellent source of valuable terpenoid, cannabinoid and polyphenolic antioxidant agents, and further research should be focused on their utilization in dietary supplements and functional foods.

Influences of chemotype and parental genotype on metabolic fingerprints of tansy plants uncovered by predictive metabolomics

Intraspecific plant chemodiversity shapes plant-environment interactions. Within species, chemotypes can be defined according to variation in dominant specialised metabolites belonging to certain classes. Different ecological functions could be assigned to these distinct chemotypes. However, the roles of other metabolic variation and the parental origin (or genotype) of the chemotypes remain poorly explored. Here, we first compared the capacity of terpenoid profiles and metabolic fingerprints to distinguish five chemotypes of common tansy (Tanacetum vulgare) and depict metabolic differences. Metabolic fingerprints captured higher variation in metabolites while preserving the ability to define chemotypes. These differences might influence plant performance and interactions with the environment. Next, to characterise the influence of the maternal origin on chemodiversity, we performed variation partitioning and generalised linear modelling. Our findings revealed that maternal origin was a higher source of chemical variation than chemotype. Predictive metabolomics unveiled 184 markers predicting maternal origin with 89% accuracy. These markers included, among others, phenolics, whose functions in plant-environment interactions are well established. Hence, these findings place parental genotype at the forefront of intraspecific chemodiversity. We recommend considering this factor when comparing the ecology of various chemotypes. Additionally, the combined inclusion of inherited variation in main terpenoids and other metabolites in computational models may help connect chemodiversity and evolutionary principles.

The non-specific lipid transfer protein McLTPII.9 of Mentha canadensis is involved in peltate glandular trichome density and volatile compound metabolism.

Mentha canadensis L. is an important spice crop and medicinal herb with high economic value. The plant is covered with peltate glandular trichomes, which are responsible for the biosynthesis and secretion of volatile oils. Plant non-specific lipid transfer proteins (nsLTPs) belong to a complex multigenic family involved in various plant physiological processes. Here, we cloned and identified a non-specific lipid transfer protein gene (McLTPII.9) from M. canadensis, which may positively regulate peltate glandular trichome density and monoterpene metabolism. McLTPII.9 was expressed in most M. canadensis tissues. The GUS signal driven by the McLTPII.9 promoter in transgenic Nicotiana tabacum was observed in stems, leaves, and roots; it was also expressed in trichomes. McLTPII.9 was associated with the plasma membrane. Overexpression of McLTPII.9 in peppermint (Mentha piperita. L) significantly increased the peltate glandular trichome density and total volatile compound content compared with wild-type peppermint; it also altered the volatile oil composition. In McLTPII.9-overexpressing (OE) peppermint, the expression levels of several monoterpenoid synthase genes and glandular trichome development-related transcription factors-such as limonene synthase (LS), limonene-3-hydroxylase (L3OH), geranyl diphosphate synthase (GPPS), HD-ZIP3, and MIXTA-exhibited varying degrees of alteration. McLTPII.9 overexpression resulted in both a change in expression of genes for terpenoid biosynthetic pathways which corresponded with an altered terpenoid profile in OE plants. In addition, peltate glandular trichome density was altered in the OE plants as well as the expression of genes for transcription factors that were shown to be involved in trichome development in plants.

Terpenoid VOC profiles and functional characterization of terpene synthases in diploid and tetraploid cytotypes of Chrysanthemum indicum L

Chrysanthemum indicum L. is a valuable medicinal plant with diploid and tetraploid forms that are widely distributed in central and southern China, and it contains abundant volatile organic compounds (VOCs). Despite the discovery of some terpene synthase (TPS) in C. indicum (i.e., CiTPS) in previous studies, many TPSs and their corresponding terpene biosynthesis pathways have yet to be discovered. In the present study, terpenoid VOCs in different tissues from two cytotypes of C. indicum were analyzed. We identified 52 types of terpenoid VOCs and systematically investigated the content and distribution of these compounds in various tissues. The two cytotypes of C. indicum exhibited different volatile terpenoid profiles. The content of monoterpenes and sesquiterpenes in the two cytotypes showed an opposite trend. In addition, four full-length candidate TPSs (named CiTPS5–8) were cloned from Ci-GD4x, and their homologous TPS genes were screened based on the genome data of Ci-HB2x. These eight TPSs displayed various tissue expression patterns and were discovered to produce 22 terpenoids, 5 of which are monoterpenes and 17 are sesquiterpenes. We further proposed corresponding terpene synthesis pathways, which can enable the establishment of an understanding of the volatile terpenoid profiles of C. indicum with different cytotypes. This knowledge may provide a further understanding of germplasm in C. indicum and may be useful for biotechnology applications of Chrysanthemum plants.

Volatile terpenoid profiling in gin and beer – A targeted approach

Volatile terpenoid profiling in gin and beer – A targeted approach

Essential Oils from Apiaceae, Asteraceae, Cupressaceae and Lamiaceae Families Grown in Serbia: Comparative Chemical Profiling with In Vitro Antioxidant Activity.

The aim of the present study was to investigate the chemical profile and antioxidant activity of essential oils obtained from the most commonly grown plant species in Serbia. Aromatic and medicinal plants from Lamiaceae (Mentha x Piperita, Ocimum basilicum, Origanum majorana, Origanum vulgare, Salvia officinalis, Satureja hortensis, Satureja montana and Thymus vulgaris), Asteraceae (Ehinacea purpurea and Matricaria chamomilla), Apiaceae (Anethum graveolens, Carum carvi, Foeniculum vulgare, Petroselinum crispum and Pimpinella anisum) and Cupressaceae (Juniperus comunis) were selected as raw material for essential oils (EOs)' isolation. Hydrodistillation (HD) was used for the isolation of EOs while they were evaluated in terms of yield and terpenoid profiles by GC-MS. In vitro radical scavenging DPPH and ABTS+ radical activities were carried out for all EOs. Finally, a principal component analysis (PCA) was performed with the experimental results of the composition and antioxidant activity of the EOs, which showed a clear distinction between the selected plant species for the aforementioned responses. This work represents a screening tool for the selection of other EO candidates for further processing by emerging extraction techniques and the use of EOs as natural additives for meat products.

Phytochemical Study of Euphorbia turcomanica Boiss.

The attraction to the Euphorbia genus, with its remarkable diversity in species, is due to its variety of chemical compositions. Euphorbia turcomanica is one of the species of the spurge family growing wildly in Iran. This research aims to investigate the presence of secondary metabolites, specially terpenoid compounds, in terms of structural determination. Samples of E. tucomanica were finely powdered and macerated with acetone/dichloromethane 2:1. Repeated column chromatography using silica gel, MPLC, and HPLC methods followed by the analysis of data obtained from spectroscopic means was carried out to purify and identify the terpenoid constituents. The chemical structures of nine known terpenoids were determined for the first time from E. turcomanica during this study. Loliolide (1), a monoterpene, and eight steroids and triterpenes, including simiarenol (2), isomultiflorenol (3), cycloart-25-ene-3β,24-diol (4), β-sitosterol (5), cycloart-23-ene-3β,25-diol (6), 3α, 11α-dihydroxyurs-12-ene (7), 3β, 24β, 25-trihydroxycycloartane (8), and 7α-hydroxystigmasterol (9) were isolated and identified. E. tucomanica, with a rich terpenoid profile, can be one of the valuable and economic sources providing compounds for drug development.

Aroma and terpenoid profile of Chardonnay clone 809: from berry to sparkling wine

Clone selection is a viticultural practice applied to evaluate the behaviour and agronomic and flavour characteristics of a material in a determined environment to obtain wines with different aromatic attributes. The clones of the cultivar Chardonnay comprise aromatic and non-aromatic clones that can provide different sensorial qualities. This work aimed to evaluate the volatile compounds formed in clone 809 (aromatic) to produce a more aromatic sparkling wine in Serra da Mantiqueira (southeast Brazil) and compare it to clone 76 (non-aromatic) from the berry to the wine. The vineyards are in Caldas (Minas Gerais – Brazil) at the Agronomical Research Centre of Minas Gerais (EPAMIG), and the vinification was performed using the Champenoise method (18 months in sur lie). HS-SPME/GC-MS identified free volatile compounds in two consecutive seasons, 2017 and 2018, in grape, musts, base wines and sparkling wines. The number and abundance of monoterpenoid compounds in clone 809 were statistically significant compared to clone 76 for all sample steps. Multivariate and principal component analyses (PCA) made it possible to differentiate the base and sparkling wines by the clones in both vintages for the second component (16.8 %). Furthermore, the sparkling wines of clone 809 were discriminated for the third component (15.1 %) by the monoterpenes: α-terpineol, linalool, ß-myrcene, hotrienol, nerol oxide, and limonene. The data suggest that Chardonnay clone 809 can contribute to obtaining a sparkling wine with an additional floral and sweet character.

Genetic Manipulation of Biosynthetic Pathways in Mint.

In recent years, the study of aromatic plants has seen an increase, with great interest from industrial, academic, and pharmaceutical industries. Among plants attracting increased attention are the Mentha spp. (mint), members of the Lamiaceae family. Mint essential oils comprise a diverse class of molecules known as terpenoids/isoprenoids, organic chemicals that are among the most diverse class of naturally plant derived compounds. The terpenoid profile of several Mentha spp. is dominated by menthol, a cyclic monoterpene with some remarkable biological properties that make it useful in the pharmaceutical, medical, cosmetic, and cleaning product industries. As the global market for Mentha essential oils increases, the desire to improve oil composition and yield follows. The monoterpenoid biosynthesis pathway is well characterised so metabolic engineering attempts have been made to facilitate this improvement. This review focuses on the Mentha spp. and attempts at altering the carbon flux through the biosynthetic pathways to increase the yield and enhance the composition of the essential oil. This includes manipulation of endogenous and heterologous biosynthetic enzymes through overexpression and RNAi suppression. Genes involved in the MEP pathway, the menthol and carvone biosynthetic pathways and transcription factors known to affect secondary metabolism will be discussed along with non-metabolic engineering approaches including environmental factors and the use of plant growth regulators.

Amino acid, sugar, phenolic, and terpenoid profiles are capable of distinguishing Citrus tristeza virus infection status in citrus cultivars: Grapefruit, lemon, mandarin, and sweet orange.

Citrus tristeza virus (CTV) is the most severe viral disease for citrus production. Many strains of CTV have been characterized and their symptomology widely varies, ranging from asymptomatic or mild infections to severe symptomology that results in substantial yield loss or host death. The capacity of the different CTV strains to affect the biochemistry of different citrus species has remained largely unstudied, despite that associated metabolomic shifts would be relevant toward symptom development. Thus, amino acid, sugar, phenolic, and terpenoid levels were assessed in leaves of healthy and CTV-infected grapefruit, lemon, mandarin, and two different sweet orange cultivars. Both mild [VT-negative (VT-)] and severe [VT-positive (VT+)] CTV genotype strains were utilized. When looking at overall totals of these metabolite classes, only amino acid levels were significantly increased by infection of citrus with severe CTV strains, relative to mild CTV strains or healthy plants. No significant trends of CTV infection on summed amounts of all sugar, phenolic, or terpenoid compounds were observed. However, individual compound levels were affected by CTV infections. Subsequent canonical discriminant analysis (CDA) that utilized profiles of individual amino acids, terpenoids, or phenolics successfully distinguished leaf samples to specific citrus varieties and identified infection status with good accuracy. Collectively, this study reveals biochemical patterns associated with severity of CTV infections that can potentially be utilized to help identify in-field CTV infections of economic relevance.

Terpenoid biosynthesis in Dendrobium officinale: Identification of (E)-β-caryophyllene synthase and the regulatory MYB genes

Dendrobium officinale Kimura et Migo is a well-known medicinal plant producing terpenoid alkaloids as the main medicinal constituents. D. officinale also produces many volatile terpenoids, including monoterpenes and sesquiterpenes. However, little is known about the biosynthetic and regulatory mechanisms underlying terpenoid biosynthesis in this plant. This study first analyzed the terpenoid profiles of D. officinale PLBs treated with lovastatin (LOV) and fosmidomycin (FOS), two chemical inhibitors of terpenoid pathways, and produced corresponding transcriptomes. The volatile products of PLBs were mainly sesquiterpenes, and the contents of (E)-β-caryophyllene and α-humulene were decreased by 42.05% and 27.27%, respectively, after LOV treatment. A candidate sesquiterpene synthase gene identified from the transcriptome and terpene metabolic profiling as (E)-β-caryophyllene synthase (DoECS), localized in the cytoplasm. The emission of (E)-β-caryophyllene decreased by 26.70% in DoECS RNA interference PLBs compared with the control group. Based on co-expression analysis, four MYB genes (DoMYB26, DoMYB29, DoMYB31 and DoMYB32) were involved in the transcriptional regulation of DoECS. Yeast one-hybrid assays and fluorometric GUS assay provided evidence that DoMYB26 and DoMYB31 positively regulated the expression of DoECS, while DoMYB29 negatively regulated the expression of DoECS. In summary, this study produced novel knowledge about the catalytic and regulatory mechanisms responsible for the biosynthesis of volatile terpenoids in D. officinale.

Foodomics for agroecology: Differentiation of volatile profile in mint (Mentha × gracilis Sole) from permaculture, organic and conventional agricultural systems using HS-SPME/GC–MS

In the present study, foodomics approach was employed to investigate changes in the metabolism from the volatile terpenoids profile of mint(Mentha × gracillis Sole)from conventional, organic and permaculture (a type of agroecological agriculture system) farms using headspace solid-phase microextraction (HS-SPME) associated to gas chromatography coupled to mass spectrometry (GC–MS) and chemometric tools. The discrimination among the three types of mint was successfully achieved and demonstrated evidence of ecological interaction impact in the food metabolism. The agroecological mint presented as differential compounds: α-terpineol, bornyl formate, cis-carvyl propionate, cis-carveol, camphor, dihydrocarvyl acetate, dihydrocarveol, karahanaenone, nonanal, 3-octyl acetate, and trans-3-hexenyl-2 methylbutyrate. While organic and conventional mint presented as differential compounds: α-cedrene, β -pinene, γ-muurolene, δ-cadinene, germacrene, terpinolene, and elemol. The majority of differential metabolites from agroecological mint are oxygenated monoterpenes, which have more intense flavor and biological activities than hydrocarbons monoterpenes and sesquiterpenes found in organic and conventional mint. Furthermore, the discrimination between organic and conventional mint was effectively performed, which demonstrated different terpenoid profiles though without implying benefits for one or another agriculture system.

Fatty acid and terpenoid profile: A signature of mountain milk

The aim of this work was to verify the suitability of terpenoids and fatty acid composition to authenticate the altitude of milk production. Milk originating from lowland (L1), mountain (M1) and alpine pasture (H1 and H2) in the Aosta Valley (Northwest Italy), during the month of June, was collected at a cheese factory. The M1, H1, and H2 milk is characterised by a high content of α-linolenic, vaccenic acid and conjugated linoleic fatty acids, compared with milk from lowland pastures. Terpenoid profiles changed with altitude of grazing. The leave-one-out cross-validation, using altitude of grazing as a grouping, and C14:0 iso, anteisoC17:0, C18:1 trans-11, C18:2 trans-9, trans-12 and cis-ocimene as discriminant variables, gave a correct classification rates of 98% of milk samples to origin of production.

Determination of Terpenoid Profile in Dry Cannabis Flowers and Extracts Obtained from Different Cannabis Varietes

Aim: The aim of this study was to determine the terpenoid profile in dried cannabis flowers obtained from different varieties of cannabis plant and in cannabis extracts in order to investigate quantity of terpenes lost during extraction and purification process.
 Methods: GC/MS method for determination of terpenes was verified. The concentration of terpenes was determined in dry flowers as raw material and in decarboxylated and distillated cannabis extracts, using the same GC/MS analytical method. The extraction was performed using 96% ethanol as a solvent.
 Results: The obtained results indicate that dry cannabis flowers from different cannabis plant can be distinguished only by their terpenoid profile. The use of standardized cannabis-based extracts can be confirmed by determination of terpenoid profile. The purification process of the cannabis extracts removes terpenes. The percentage of major terpen beta-Myrcene decreased from 68% in dry flower to 15% in decarboxylated and, 1.9% in distillated cannabis oil after purification. The percentage of second major terpene alpha-Pinene decreased from 15% in dry flower to 5% in decarboxylated and, 0.7% in distillated cannabis oil after purification.
 Conclusion: Terpenes act synergistically with cannabinoids. Following the monograph for quality testing of cannabis extracts in the German Pharmacopoeia, the purification process is necessary to achieve a final concentration of cannabinoids (Tetrahydrocannabinol) of more than 95% in the final active pharmaceutical ingredient. The purification process removes terpenes that have proven synergistically pharmacological effects with cannabinoids.