Terpenoid Family Research Articles

Changing biosynthesis of terpenoid percursors in rice through synthetic biology.

Many highly valued chemicals in the pharmaceutical, biotechnological, cosmetic, and biomedical industries belong to the terpenoid family. Biosynthesis of these chemicals relies on polymerization of Isopentenyl di-phosphate (IPP) and/or dimethylallyl diphosphate (DMAPP) monomers, which plants synthesize using two alternative pathways: a cytosolic mevalonic acid (MVA) pathway and a plastidic methyleritritol-4-phosphate (MEP) pathway. As such, developing plants for use as a platform to use IPP/DMAPP and produce high value terpenoids is an important biotechnological goal. Still, IPP/DMAPP are the precursors to many plant developmental hormones. This creates severe challenges in redirecting IPP/DMAPP towards production of non-cognate plant metabolites. A potential solution to this problem is increasing the IPP/DMAPP production flux in planta. Here, we aimed at discovering, understanding, and predicting the effects of increasing IPP/DMAPP production in plants through modelling. We used synthetic biology to create rice lines containing an additional ectopic MVA biosynthetic pathway for producing IPP/DMAPP. The rice lines express three alternative versions of the additional MVA pathway in the plastid, in addition to the normal endogenous pathways. We collected data for changes in macroscopic and molecular phenotypes, gene expression, isoprenoid content, and hormone abundance in those lines. To integrate the molecular and macroscopic data and develop a more in depth understanding of the effects of engineering the exogenous pathway in the mutant rice lines, we developed and analyzed data-centric, line-specific, multilevel mathematical models. These models connect the effects of variations in hormones and gene expression to changes in macroscopic plant phenotype and metabolite concentrations within the MVA and MEP pathways of WT and mutant rice lines. Our models allow us to predict how an exogenous IPP/DMAPP biosynthetic pathway affects the flux of terpenoid precursors. We also quantify the long-term effect of plant hormones on the dynamic behavior of IPP/DMAPP biosynthetic pathways in seeds, and predict plant characteristics, such as plant height, leaf size, and chlorophyll content from molecular data. In addition, our models are a tool that can be used in the future to help in prioritizing re-engineering strategies for the exogenous pathway in order to achieve specific metabolic goals.

Chemical Constituents and Their Production in Mexican Oaks (Q. Rugosa, Q. Glabrescens and Q. Obtusata).

Mexico is considered one of the main regions of diversification of the genus Quercus (oaks). Oak species are one of the most important tree groups, particularly in temperate forests, due to its diversity and abundance. Some studies have shown that oak contains specialized metabolites with medicinal importance. In this work, the acetonic extract from leaves of three Mexican oaks (Quercus rugosa, Q. glabrescens, and Q. obtusata) was separated using thin-layer chromatography and column chromatography. Chemical identification of the major compounds was determined using high-performance liquid chromatography and nuclear magnetic resonance. Nineteen compounds were identified, three belonging to the terpenoid family (ursolic acid, β−amyrin, and β−sitosterol) and 16 from the phenolic family. Of the isolated compounds, seven are new reports for oak species (scopoletin, ursolic acid, β-amyrin, luteolin−7−O−glucoside, kaempferol−3−O−sophoroside, kaempferol−3−O−glucoside, and kaempferol−3−O−sambubioside). More compounds were identified in Q. rugosa followed by Q. glabrescens and then Q. obtusata. The characterization of specialized metabolites in oak species is relevant, from both phytocentric and anthropocentric perspectives.

Optimal NaCl Medium Enhances Squalene Accumulation in Thraustochytrium sp. ATCC 26185 and Influences the Expression Levels of Key Metabolic Genes.

Squalene, a natural lipid of the terpenoid family, is well-recognized for its roles in regulating cholesterol metabolism, preventing tumor development, and improving immunity. For large-scale squalene production, the unicellular marine protists—thraustochytrids—have shown great potential. However, the growth of thraustochytrids is known to be affected by salt stress, which can eventually influence the squalene content. Here, we study the effects of an optimal concentration of NaCl on the squalene content and transcriptome of Thraustochytrium sp. ATCC 26185. Under the optimal culture conditions (glucose, 30 g/L; yeast extract, 2.5 g/L; and NaCl, 5 g/L; 28°C), the strain yielded 67.7 mg squalene/g cell dry weight, which was significantly greater than that (5.37 mg/g) under the unoptimized conditions. NaCl was determined as the most significant (R = 135.24) factor for squalene production among glucose, yeast extract, and NaCl. Further comparative transcriptomics between the ATCC 26185 culture with and without NaCl addition revealed that NaCl (5 g/L) influences the expression of certain key metabolic genes, namely, IDI, FAS-a, FAS-b, ALDH3, GS, and NDUFS4. The differential expression of these genes possibly influenced the acetyl-CoA and glutamate metabolism and resulted in an increased squalene production. Through the integration of bioprocess technology and transcriptomics, this report provides the first evidence of the possible mechanisms underscoring increased squalene production by NaCl.

Molecular Dynamics Simulations of Essential Oil Ingredients Associated with Hyperbranched Polymer Drug Carriers.

Our work concerns the study of four candidate drug compounds of the terpenoid family, found as essential oil ingredients in species of the Greek endemic flora, namely carvacrol, p-cymene, γ-terpinene, and thymol, via the simulation method of molecular dynamics. Aquatic solutions of each compound, as well as a solution of all four together in realistic (experimental) proportions, are simulated at atmospheric pressure and 37 °C using an OPLS force field combined with TIP3P water. As verified, all four compounds exhibit a strong tendency to phase-separate, thereby calling for the use of carrier molecules as aids for the drug to circulate in the blood and enter the cells. Systems of two such carrier molecules, the hyperbranched poly(ethylene imine) (HBPEI) polyelectrolyte and hyperbranched polyglycerol (HPG), are examined in mixtures with carvacrol, the most abundant among the four compounds, at a range of concentrations, as well as with all four compounds present in natural proportions. Although a tendency of the terpenoids to cluster separately persists at high concentrations, promising association effects are observed for all drug–polymer ratios. HBPEI systems tend to form diffuse structures comprising small mixed clusters as well as freely floating polymer and essential oil molecules, a finding attributed to the polymer–polymer electrostatic repulsions, which here are only partially screened by the counterions. On the other hand, the electrically neutral HPG molecules cluster together with essential oil species to form a single nanodroplet. Currently, terpenoid–polymer clusters near lipid bilayer membranes are being studied to determine the propensity of the formed complexes to enter cell membranes.

Cloning and Characterization of 1,8-Cineole Synthase (SgCINS) Gene From the Leaves of Salvia guaranitica Plant.

Monoterpenes are one of the most common groups belonging to the terpenoid family, with a C10 structure comprising of two isoprene units. Most of monoterpenes are volatile plant compounds, and they act as signaling molecules between plants and the environment, particularly as defensive compounds against herbivores and pathogens. In this study, 1,8-cineole synthase (SgCINS) gene was identified and cloned from the leaves of Salvia guaranitica plant. To examine the role of SgCINS in insect resistance, we transformed and expressed this gene into tobacco leaves. The metabolic analysis revealed that the production of various types and amount of terpenoid was increased and decreased in SgCINS overexpression and control lines, respectively, suggesting that overexpressing SgCINS in transgenic tobacco plants lead to an increase in the production of various types of terpenoids and other phytochemical compounds. These results indicated why transgenic tobacco was highly resistant against cotton worm than the highly susceptible control plants. Our results demonstrate that the SgCINS gene can play an important role in plants against cotton worm insect attack, and pave the way for using terpenoids genes for improving resistance to insect attack in higher plants.

Pre-fermentative maceration with SO2 enhanced the must aromatic composition

SO2 is the additive most used in winemaking, due to its antioxidant and antiseptic activities. There have been several studies focused on these activities; however, there are hardly any studies that address its role as extractant. Therefore, the objective of this work was to study the effect of SO2 on the must volatile composition after a pre-fermentation maceration. The results showed that the influence of SO2 on the extraction of the two most important families of varietal compounds, terpenoids and C13 norisoprenoids, was different: the content of total terpenoids increased by 95%, while for the total of C13 norisoprenoids no significant differences were observed. The other three families of volatile compounds studied, benzenoid compounds, esters, and C6 compounds, were found in greater quantity in the samples with SO2, increasing by 51%, 164%, and 45%, respectively. Consequently, SO2 can enhance the must aromatic composition of neutral varieties, such as Tempranillo.

Contribution of Volatile Odorous Terpenoid Compounds to Aged Cognac Spirits Aroma in a Context of Multicomponent Odor Mixtures.

Cognac spirit aromas result from the presence of a wide variety of volatile odorous compounds associated with the modalities of spirit distillation and aging. However, very few studies have been carried out on aging notes of Cognac spirits. An HPLC fractionation approach was used in order to evidence fractions of interest recalling the specific aromatic nuances of aged Cognac. Then, a GC-O/MS analysis of the selected fractions allowed one to detect odorous zones and identify several volatile compounds. Among them, various well-known volatile compounds representative of the terpenoid family were highlighted, such as geraniol, α-terpinene, nerol, α-terpineol, 1,8-cineole (eucalyptol) and, particularly, piperitone, santalol, and α-campholenal, which have not previously been cited in Cognac. These compounds were quantitated and their detection thresholds were determined. Geraniol, α-terpinene, α-terpineol, and 1,8-cineole concentrations increased while spirits were more aged, while nerol tends to decrease. A sensory contribution of terpenes was observed through perceptual synergic effects, along with β-damascenone and whisky lactone.

Inhibition of jasmonate-mediated plant defences by the fungal metabolite higginsianin B.

Infection of Arabidopsis thaliana by the ascomycete fungus Colletotrichum higginsianum is characterized by an early symptomless biotrophic phase followed by a destructive necrotrophic phase. The fungal genome contains 77 secondary metabolism-related biosynthetic gene clusters, whose expression during the infection process is tightly regulated. Deleting CclA, a chromatin regulator involved in the repression of some biosynthetic gene clusters through H3K4 trimethylation, allowed overproduction of three families of terpenoids and isolation of 12 different molecules. These natural products were tested in combination with methyl jasmonate, an elicitor of jasmonate responses, for their capacity to alter defence gene induction in Arabidopsis. Higginsianin B inhibited methyl jasmonate-triggered expression of the defence reporter VSP1p:GUS, suggesting it may block bioactive jasmonoyl isoleucine (JA-Ile) synthesis or signalling in planta. Using the JA-Ile sensor Jas9-VENUS, we found that higginsianin B, but not three other structurally related molecules, suppressed JA-Ile signalling by preventing the degradation of JAZ proteins, the repressors of jasmonate responses. Higginsianin B likely blocks the 26S proteasome-dependent degradation of JAZ proteins because it inhibited chymotrypsin- and caspase-like protease activities. The inhibition of target degradation by higginsianin B also extended to auxin signalling, as higginsianin B treatment reduced auxin-dependent expression of DR5p:GUS. Overall, our data indicate that specific fungal secondary metabolites can act similarly to protein effectors to subvert plant immune and developmental responses.

Insights into the Mechanism and Enantioselectivity in the Biosynthesis of Ergot Alkaloid Cycloclavine Catalyzed by Aj_EasH from Aspergillus japonicus.

Cycloclavine is a complex ergot alkaloid containing an unusual cyclopropyl moiety, which has a wide range of biological activities and pharmaceutical applications. The biosynthesis of cycloclavine requires a series of enzymes, one of which is a nonheme FeII/α-ketoglutarate-dependent (aKG) oxidase (Aj_EasH). According to the previous proposal, the cyclopropyl ring formation catalyzed by Aj_EasH follows an unprecedented oxidative mechanism; however, the reaction details are unknown. In this article, on the basis of the recently obtained crystal structure of Aj_EasH (EasH from Aspergillus japonicas), the reactant models were built, and the reaction details were investigated by performing QM-only and combined QM and MM calculations. Our calculation results reveal that the biosynthesis of cyclopropyl moiety involves a radical intermediate rather than a carbocationic or carbanionic intermediate as in the biosynthesis of terpenoid family. The iron(IV)-oxo first abstracts a hydrogen atom from the substrate to trigger the reaction, and then the generated radical intermediate undergoes ring rearrangement to form the fused 5-3 ring system of cycloclavine. On the basis of our calculations, the absolute configuration of the cycloclavine catalyzed by Aj_EasH from Aspergillus japonicus should be (5R,8R,10R), which is different from the product isolated from Ipomoea hildebrandtii (5R,8S,10S). Residues at the active site play an important role in substrate binding, ring rearrangement, and enantioselectivity.

Phytochemistry and neuroprotective effects of Eclipta alba (L.) Hassk.

Eclipta alba (L.) Hassk. or Eclipta prostrata (Linn.) or Eclipta erecta (Linn.) is an herbaceous plant well known in Asian as African traditional medicines. These extracts are used in traditional medicine for treatment of microbial diseases and certain metabolic disorders. This review aimed to investigate phytochemical profile and neuroprotective effects of E. alba (L.) Hassk. Several compounds belonging to the families of phenolics, alkaloids, terpenoids and polysaccharides have been isolated, identified or characterized from E. alba extracts. This plant has a diverse neuropharmacological profile. Thus, its extract improves cognitive deficits and also attenuated epileptic seizures. Phytomolecules implicated in these potentials are Eclalbasaponin II and luteolin, respectively. This document updates isolated and identified organic compounds from the extracts of E. alba and reviews their neuropharmacological activities.

Penentuan Nilai Sun Protection Factor secara In Vitro pada Ekstrak Etanol Akar Kalakai (Stenochlaena palustris Bedd) dengan Metode Spektrofotometer UV-Vis

The sunscreens or Sun Protecting Agent is the materials protecting the skin from the exposure to ultraviolet (UV) rays. Amount of Sun Protection Factor shows how many times the subject skin protection can be multiplied under the sun rays without erythema occurs. Kalakai contains some bio-active substance like phenol, flavonoid, alkaloid and terpenoid family which are proven as a very effective antioxidant. The purpose of this research was to determine the concentration of ethanol extract of kalakai root which has the potential as a sunscreen based on the Sun Protection Factor (SPF) parameter. Determination of SPF amount was carried out in vitro by UV-VIS spectrophotometer at a concentration of 50 ppm, 100 ppm, 150 ppm, 200 ppm, 250 ppm, 300 ppm, and 350 ppm with a wavelength of 290-320 nm. The result showed that kalakai root ethanol extract indicated potential as a sunscreen at a concentration of 300 ppm and 350 ppm obtained SPF values respectively 11 and 14 classified as sunscreens with extreme ability levels.

Occurrence and Biological Activities of Phenylpropionyl Iridoids.

As noteworthy members of the terpenoid family, iridoids are a group of natural compounds that have a cyclopentane ring and hemiacetal structural features. Among them, phenylpropionyl iridoids- widely present in Bignoniaceae, Rubiaceae, Scrophulariaceae, Verbenaceae, and Oleaceae- were discovered and extracted from plants mainly between the 1980s and 2000s. Because of their unique structural features and complex stereo configurations, these compounds exhibit numerous desirable biological activities such as neuroprotective, anti-inflammatory, antimicrobial, and antitumour activities; they have received increasing interest recently. This review summarises progress in research on phenylpropionyl iridoids, including their occurrence and biological activities.

Sesquiterpene Production in Methyl Jasmonate-Induced Persicaria minor Cell Suspension Culture

Sesquiterpenes are a three-isoprene unit compounds which belong to terpenoid family of secondary metabolites. These volatile compounds are one of the major constituents of essential oils in plants and plays major roles in plant signaling of defense mechanism. The effects of methyl jasmonate (MeJA) concentrations (100 and 200 μM) on the production of sesquiterpene compounds after incubation period for 1, 3, and 6 days were investigated in Persicaria minor cell suspension culture. Headspace Solid-Phase Microextraction (HS-SPME) method was used to absorb volatile compounds from suspension cells and liquid medium. They were then analyzed through Gas Chromatography-Mass Spectrometry (GC-MS) to identify sesquiterpene compounds. Among the 15 sesquiterpene compounds identified, α- muurolene was found in significantly higher concentration in all MeJA treated cultures. The results showed that α-muurolene was detected in the suspension cells at the highest peak area of 14.17% at 100 μM MeJA treated cultures with 3-day incubation. Analysis of liquid medium of the treatments identified secretion of α-muurolene into the culture medium, with total peak area of 0.72%. These results showed that sesquiterpene production in MeJA induced P. minor suspension culture depended on the MeJA concentration and also culture incubation period.

Terpenoids as cytotoxic compounds: A perspective

Natural products serve as safe and effective therapeutic agents for the drug discovery. Plants produce bioactive secondary metabolites such as terpenoids, alkaloids, tannins, saponins, and others, which have profusely been studied for anti-infective, anticancer, anti-inflammatory potential, and metabolic disorders to name a few. Terpenoids constitute the largest class of natural compound. The study investigates the novel cytotoxic compound of the terpenoid family isolated from different natural sources. The anticancerous activities of the compounds discussed in this review are taken from the published articles showing activity against specific cancer cell lines. The compounds exclusively belong to terpenoid family. Considering the huge potential of plant-based natural products in drug discovery program and the contribution of terpenoid in the production of anticancer compounds, it can be exploited for more reliable economical and environmentally safe bioactive molecules.

Design, Preparation and Characterization of Modular Squalene-based Nanosystems for Controlled Drug Release.

This article reviews the innovative and original concept the "squalenoylation", a technology allowing the formulation of a wide range of drug molecules (both hydrophilic and lipophilic) as nanoparticles. The "squalenoylation" approach is based on the covalent linkage between the squalene, a natural and biocompatible lipid belonging to the terpenoid family, and a drug, in order to increase its pharmacological efficacy. Fundamentally, the dynamically folded conformation of squalene triggers the resulting squalene-drug bioconjugates to self-assemble as nanoparticles of 100-300 nm. In general, these nanoparticles showed long blood circulation times after intravenous administration and improved pharmacological activity with reduced side effects and toxicity. This flexible and generic technique opens exciting perspectives in the drug delivery field.

Characterization of the first naturally thermostable terpene synthases and development of strategies to improve thermostability in this family of enzymes

The terpenoid family of natural products is being targeted for heterologous microbial production as a cheaper and more reliable alternative to extraction from plants. The key enzyme responsible for diversification of terpene structure is the class‐I terpene synthase (TS), and these often require engineering to improve properties such as thermostability, robustness and catalytic activity before they are suitable for industrial use. Improving thermostability typically relies on screening a large number of mutants, as there are no naturally thermostable TSs described upon which to base rational design decisions. We have characterized the first examples of natural TSs exhibiting thermostability, which catalyse the formation of the sesquiterpene τ‐muurolol at temperatures up to 78 °C. We also report an enzyme with a k cat value of 0.95 s−1 at 65 °C, the highest k cat recorded for a bacterial sesquiterpene synthase. In turn, these thermostable enzymes were used as a model to inform the rational engineering of another TS, with the same specificity but low sequence identity to the model. The newly engineered variant displayed increased thermostability and turnover. Given the high structural homology of the class‐I TS domain, this approach could be generally applicable to improving the properties of other enzymes in this class.DatabaseModel data are available in the PMDB database under the accession number PM0080780.

Overexpression of Panax ginseng sesquiterpene synthase gene confers tolerance against Pseudomonas syringae pv. tomato in Arabidopsis thaliana.

Sesquiterpenes are an abundant group belonging to the terpenoid family, with a C15 structure comprise of three isoprene units. Many sesquiterpenes are volatile compounds and it act as chemical messenger in plant signalling, particularly in the defense mechanism against biotic and abiotic stresses. Panax ginseng Meyer is important medicinal herbs with various reported pharmacological efficacies in which its triterpenoid saponins, called ginsenosides, were mostly studied. However, there have been few studies on volatile sesquiterpenes compounds regulation on P. ginseng. As slow-growing perennial plant, P. ginseng received many kind of stresses during its cultivation. The pathogen attack is one of the most devastated perturbation for ginseng yield. Thus, we aimed to analyze P. ginseng STS gene (PgSTS) expressions in ginseng organs as well as mono-, sesquiterpenes contents from ginseng seedlings treated with elicitors. qRT-PCR and GC-MS analysis showed that two elicitors- salicylic acid (SA) and methyl jasmonate (MeJA) triggered PgSTS expression at different time points and significantly induced mono-, sesquiterpene yield. Overexpression of PgSTS in Arabidopsis also induced high terpene content and conferred tolerance against Pseudomonas syringae pv. tomato infection. These results suggested that PgSTS transcripts are involved in terpenoid biosynthesis in response to environmental stress mediated by MeJA and SA elicitors; thus, generate tolerance against pathogen attack.

Preliminary Investigation of Bioactive Compounds and Bioautographic Studies of Whole Plant Extract of Euphorbia pulcherrima on Escherichia coli, Staphylococcus aureus, Salmonella typhi, and Pseudomonas aeruginosa

The aim of this study is to carry out preliminary investigation of bioactive compounds and bioautographic studies of whole plant extract of Euphorbia pulcherrima on Escherichia coli, Staphylococcus aureus, Salmonella typhi, and Pseudomonas aeruginosa. Tukey HSD test of hierarchy for the effect of different solvents crude extract on bacterial isolates indicates the methanol extract as the most bioactive. The Tukey HSD analysis also showed that the bioactivities of the crude extracts of the various parts of Euphorbia pulcherrima were part dependent and the whole plant was the most bioactive. The ethyl acetate fraction of the methanol extract of the whole plant of Euphorbia pulcherrima has been shown in this work to contain phytochemicals which have shown remarkable activities against Escherichia coli, Staphylococcus aureus, Salmonella typhi, and Pseudomonas aeruginosa. The bioactivities against the test organisms were due to the combined effects of the compounds separated on TLC plates. Families of terpenoids, flavonoids, alkaloids, saponin, and steroids that were detected in the extracts were identified by GC-MS. The various classes of phytochemicals in the E. pulcherrima plant provided the antimicrobial potency of the plant.

Exploring the potentialities of comprehensive two-dimensional gas chromatography coupled to time of flight mass spectrometry to distinguish bivalve species: Comparison of two clam species (Venerupis decussata and Venerupis philippinarum)

Metabolomics represents an emerging topic that can be valuable in the knowledge of organism responses to different stimuli. Metabolomic studies of bivalves may reveal the constraints they are subjected to, and may help clarifying the functions most affected and the tolerance mechanisms triggered. In response to this approach, two-dimensional gas chromatography–time of flight mass spectrometry (GC×GC–ToFMS) combined with headspace solid phase microextraction (HS-SPME) was applied, for the first time to our knowledge, to the untargeted and comprehensive study of the volatile composition of clam species. Firstly, experimental parameters that influence the SPME extraction efficiency were evaluated: sample preparation mode, sample volume and SPME fiber coating. Taking into account the results from the optimization step, the metabolomic profiles were performed using 1ml of clam soft tissues homogenized with N2 and diluted in deionised water (1:2, w/v), using the PDMS/DVB fiber coating. From a total of more than 200 compounds detected per sample, 63 were tentatively identified and distributed over the chemical families of hydrocarbons, ketones, aldehydes, alcohols, and terpenoids, which seems to arise from clams own metabolism, and/or from their activity as filter-feeders. The potential of this methodology to discriminate close related species was explored by comparing the volatile profiles of Venerupis philippinarum and Venerupis decussata from Ria de Aveiro, two clams belonging to the same genus. Both species experience different environment conditions, and physiological and biochemical characteristics, which may explain the observed differentiation between their metabolic profiles. To our knowledge, this is the most detailed information available so far about clam volatile composition, which represents a valuable data for future advanced studies in the ecology, toxicology and physiology of bivalves based on clams fingerprinting.

Exploring the human urine metabolomic potentialities by comprehensive two-dimensional gas chromatography coupled to time of flight mass spectrometry

Metabolomics represents an emerging issue that can aid in the diagnosis and/or prognosis of different diseases. Metabolomic study of urine is particularly interesting as it can be on the base of the developing of new faster and non-invasive methodologies. In response to this actual trend, comprehensive two-dimensional gas chromatography-time of flight mass spectrometry (GC×GC-ToFMS) combined with headspace solid phase microextraction (HS-SPME) is applied, for the first time to our knowledge, to the untargeted and comprehensive study of the volatile composition of human urine. From a total of ca. 700 compounds detected per sample, 294 were tentatively identified and distributed over the chemical families of hydrocarbons, amines, amides, esters, ketones, aldehydes, alcohols, carboxylic acids, ethers, nitriles, halides, sulfides, thiols, terpenoids, and heterocyclic compounds. To our knowledge, this is the most complete information available so far about whole human urine volatile composition, which represents a valuable data for future advanced studies in the clinical field based on urine fingerprinting. Relevant SPME and GC×GC parameters were considered. Complex sample characterization of human urine is significantly simplified due to the structured GC×GC chromatogram that produces distinct spaces for metabolite chemical families. Furthermore, the potential of this methodology in health related applications was explored by comparing the urinary volatile profiles between smoker (high-risk population for lung cancer) vs. non-smoker adults, focusing on metabolites related to oxidative stress (aliphatic alkanes and aldehydes). In spite of the small sample numbers considered, the results suggest that the urinary volatile profiles may be useful for differentiating subjects with different physiological conditions, thus making it worth to further explore its diagnostic potential.