Primjena mikroekstrakcije vršnih para na čvrstoj fazi kao preparativni pristup za plinsku kromatografiju sa spektrometrijom masa

Reviewed in brief are the selected results of the application of headspace solid-phase microextraction as a preparative approach for gas chromatography – mass spectrometry (HS-SPME/GC-MS) for natural organic compounds research at the University of Split, Faculty of Chemistry and Technology. A wide variety of headspace compounds from different natural sources has been identified: lower aliphatic compounds (e.g., C5- and C6-compounds), aromatic compounds, monoterpenes (e.g., linalool derivatives (oxides, anhydro-oxides, epoxides), hotrienol), sesquiterpenes (e.g., eudesmol isomers, hydrocarbons), and C9- and C13-norisoprenoids (e.g., 3,4-dihydro-3-oxoedulan, 4-oxoisophorone, trans-β-damascenone). These compounds are important phytochemicals as flavour/fragrance compounds, chemical markers of the botanical origin or others (e.g., allelochemicals, pheromones, or acaricide residue). This work is licensed under a Creative Commons Attribution 4.0 International License.Ukratko su prikazani odabrani rezultati primjene mikroekstrakcije vršnih para na čvrstoj fazi kao preparativnog pristupa plinskoj kromatografiji – spektrometriji masa (HS-SPME/GC-MS) za istraživanje prirodnih spojeva, koje je provedeno na Kemijsko-tehnološkom fakultetu Sveučilištu u Splitu. Velika raznolikost spojeva vršnih para istražena je iz različitih prirodnih izvora: niži alifatski spojevi (npr. C5- i C6-spojevi), aromatski spojevi, monoterpeni (npr. derivati linaloola (oksidi, anhidro-oksidi, epoksidi), hotrienol), seskviterpeni (npr. eudezmol izomeri, ugljikovodici) te C9- i C13-norizoprenoidi (npr. 3,4-dihidro-3-oksoedulan, 4-oksoisoforon, trans-β-damascenon). Ti su spojevi važne fitokemikalije kao spojevi aroma/mirisa, kemijski marker botaničkog podrijetla ili drugo (npr. aleokemikalije, feromoni ili ostatci akaricida). Ovo djelo je dano na korištenje pod licencom Creative Commons Imenovanje 4.0 međunarodna

Volatile Compounds Identified in Traditional Croatian Cow’s and Goat’s Cheeses Matured in Lambskin Sack Determined by Gc-Ms Analysis

Cheeses ripened in animal skin sacks belong to traditional cheeses that are strongly connected to the culture and history of the country from which they originate. Their main specificity is anaerobic ripening in an animal skin sack. The aim of this study was to determine the volatile compounds of Croatian cheeses in a sack of lamb skin produced from either raw cow’s or goat’s milk and relate them to the aroma of these traditional cheeses. Volatile compounds were extracted by headspace solid-phase microextraction (HS-SPME) and ultrasonic solvent extraction (USE) and analysed by gas chromatography-mass spectrometry (GC-MS). A total of 32 volatiles were identified in the cheese samples, including 12 carboxylic acids, 8 esters, 6 alcohols, 3 ketones, 2 hydrocarbons and 1 aldehyde. In the samples obtained by HS-SPME, the fatty acids and alcohols were the most abundant, while in the samples obtained by USE, the fatty acids were the most abundant

BioProspecting of the Adriatic Sea: A Review of Recent Studies of Volatile Organic Compounds

Istraživanja predstavljena u ovom preglednom radu doprinose poznavanju isparljivih organskih spojeva iz morskih organizama, posebno iz makroalgi Jadranskog mora. Za izolaciju isparljivih spojeva iz svježih i/ili na zraku osušenih uzoraka primijenjene su komplementarne metode: mikroekstrakcija vršnih para na čvrstoj fazi (HS-SPME) i/ili hidrodestilacija (HD). Dobiveni izolati analizirani su vezanim sustavom plinske kromatografije-spektrometrije masa (GC-MS). U vršnim parama i isparljivim uljima makroalgi Halopteris filicina, Flabellia petiolata, Dictyota dichotoma, Taonia atomaria, Padina pavonica, Codium bursa i morskoj cvjetnici Posidonia oceanica pronađeno je mnoštvo različitih spojeva, uglavnom seskviterpena, diterpena i alifatskih spojeva. Alifatski spojevi male molekulske mase, poput dimetil-sulfida (DMS), tribrommetana, 1-jodpentana, 3-metilbut-2-enala, heksanala i pent-1-en-3-ona, nađeni su u kemijskim profilima vršnih para. Također su identificirani i alifatski C8- i C11-spojevi (npr. oktan-l-ol, oktanal, okt-1-en-3-ol, 6-metilhept-5-en-2-on, fukoseraten i diktioptereni). Zajedno s višim acikličkim ugljikovodicima, također pronađenim u vršnim parama (npr. heptadekan), u ispitivanim isparljivim uljima identificirani su i viši alifatski alkoholi (npr. (Z)-oktadec-9-en-1-ol, heksadekan-1-ol, (Z,Z)-oktadeka-3,13-dien-1-ol) i esteri (npr. diizooktil-ftalat, dibutil-ftalat). Fenilpropanski derivati (npr. benzaldehid, benzil-alkohol) i C13-norizporenoidi (npr. α-jonon i β-jonon) također su pronađeni u isparljivim profilima, kao i mnoštvo seskviterpena (npr. germakren D, biciklogermakren, δ-kadinen, epizonaren, epibicikloseskvifelandren, β-kubeben, gleenol, (E)-β-farnezen). Isparljivi organski spojevi identificirani u morskim algama mogu poslužiti u kemotaksonomskim istraživanjima. Ovo djelo je dano na korištenje pod licencom Creative Commons Imenovanje 4.0 međunarodna.Studies presented in this review contribute to the knowledge of volatile organic compounds (VOCs) from marine organisms, especially macrolgae from the Adriatic Sea. Complementary methods were used for the isolation of VOCs from fresh and/or air-dried samples: headspace solid-phase microextraction (HS-SPME) and/or hydrodistillation (HD). The isolates were analysed by gas chromatography and mass spectrometry (GC-MS). A variety of different volatile compounds, mainly sesquiterpenes, diterpenes and aliphatic compounds were found in headspace and volatile oils of macroalgae Halopteris filicina, Flabellia petiolata, Dictyota dichotoma, Taonia atomaria, Padina pavonica, Codium bursa, and one seagrass Posidonia oceanica. Low molecular aliphatic compounds were found in the chemical profiles of headspace, such as dimethyl sulphide (DMS), tribromomethane, 1-iodopentane, 3-methylbut-2-enal, hexanal and pent-1-en-3-one. Aliphatic C8- and C11-compounds (e.g., octan-1-ol, octanal, oct-1-en-3-ol, 6-methylhept-5-en-2-on, fucoserratene and dictyopterenes) were also identified. Along with higher acyclic hydrocarbons, which are also found in headspace (e.g. heptadecane), in the investigated volatile oils higher aliphatics alcohols (e.g., (Z)-oktadec-9-en-1-ol, hexadecan-1-ol, (Z,Z)-oktadeca-3,13-dien-1-ol) and esters (e.g., diisooctyl phthalate, dibutyl phthalate) were found. Phenylpropane derivatives (e.g., benzaldehyde, benzyl alcohol) and C13-norisporenoids (e.g., α-ionone and β-ionone) were also identified in the volatile profiles, as well as an array of sesquiterpenes (e.g., germacrene D, bicyclogermacrene, δ-cadinene, epizonarene, epi-bicyclosesquiphellandrene, β-cubebene, gleenol, (E)-β-farnesene). VOCs, identified in marine algae, could be useful for chemotaxonomic studies. This work is licensed under a Creative Commons Attribution 4.0 International License

Genotyping of Leptospira spp. in wild rats leads to first time detection of L. kirshneri serovar Mozdok in Serbia

Introduction: This study aimed to investigate the prevalence and molecular characterization of Leptospira species in Belgrade, Serbia, an area where this disease is underexplored. Specifically, the study sought to employ molecular and multilocus sequence typing analyses to fill the gap in understanding the diversity and distribution of Leptospira species within the region. Methods: A comprehensive molecular analysis was conducted on kidney samples obtained from Norway rats (Rattus norvegicus) in the urban environment. The study utilized molecular diagnostic techniques including real-time PCR targeting the lipL32 gene and performing sequence-based typing schemes utilizing adk, icdA, lipL32, lipL41, rrs2, and secY genes. These methodologies were applied to ascertain the presence and characterize different Leptospira species and serovars, respectively. Results: The findings revealed the presence of two Leptospira species and three separate serovars in the Belgrade area. This study identified the presence of L. kirschneri serovar Mozdok in Serbia for the first time, a significant discovery previously undocumented in the region. This pioneering investigation sheds light on the molecular diversity and prevalence of Leptospira species in Serbia. Discussion: The study underscores the importance of employing molecular typing methods to gain insights into the epidemiology and characterization of Leptospira species. These findings significantly contribute to both local and global perspectives on leptospirosis epidemiology, providing vital insights for the development of effective control strategies and interventions. Summary: In our recent study, we explored the presence and performed molecular typing of the Leptospira species, the bacteria responsible for leptospirosis, in wild rats in Serbia. This was the first time such a study was conducted in the region. Leptospirosis is a serious disease that affects both animals and humans, often transmitted through contact with water contaminated by infected animals. Our focus was on understanding which types of Leptospira were present in these animals. Excitingly, we discovered a particular strain of Leptospira, known as L. kirshneri serovar Mozdok, for the first time in Serbia. This finding is significant because it sheds light on the presence and spread of different Leptospira serovars in Serbia. It also raises awareness about the potential health risks associated with this serovar, which was previously unknown in the area. Our work fits into a broader context of disease surveillance and public health. By identifying the types of Leptospira present in a specific region, we can better understand the risks to public health and take steps to prevent and control the spread of leptospirosis. This discovery is not just important for scientists studying infectious diseases; it has real implications for public health officials, veterinarians, and anyone concerned with preventing and treating leptospirosis. Our findings highlight the need for ongoing monitoring of Leptospira in wildlife and synanthropic fauna, to protect both animal and human health

Volatile Constituents of Aerial Parts of Capsella rubella Reut.

Three different isolates from unexplored wild-growing Brassicaceae plant Capsella rubella were investigated in order to provide more complete insight of the composition and content of volatile constituents. The analysis of volatile compounds was performed by GC-FID/MS. C. rubella essential oil, that is isolate obtained by hydrodistillation, was characterized by a high percentage of 3,4-epithiobutanenitrile (67.8 %). The main volatile compounds in autolysate, i.e. isolate obtained by solvent extraction upon endogenous enzymatic hydrolysis, were 3,4-epithiobutanenitrile (44.1 %) and ethyl isothiocyanate (29.4 %). Same as in autolysate, the main volatile constituents of hydrolysate, i.e. isolate obtained by solvent extraction upon exogenous enzymatic hydrolysis, were 3,4-epithiobutanenitrile and ethyl isothiocyanate, but with ethyl isothiocyanate as the dominating compound (57.8 %) followed by 3,4-epithiobutanenitrile (15.6 %). The results showed that qualitative and quantitative composition of C. rubella volatile constituents depends on the isolation method. But, regardless of the isolation method sulfur- and nitrogen-containing volatile compounds were quantitatively dominating constituents in all isolates. This work is licensed under a Creative Commons Attribution 4.0 International License

Traceability of Satsuma Mandarin (Citrus unshiu Marc.) Honey through Nectar/Honey-Sac/Honey Pathways of the Headspace, Volatiles, and Semi-Volatiles: Chemical Markers

Headspace solid-phase microextraction (HS-SPME) and ultrasonic solvent extraction (USE), followed by GC-MS/FID, were applied for monitoring the nectar (NE)/honey-sac (HoS)/honey (HO) pathways of the headspace, volatiles, and semi-volatiles. The major NE (4 varieties of Citrus unshiu) headspace compounds were linalool, α-terpineol, 1H-indole, methyl anthranilate, and phenylacetonitrile. Corresponding extracts contained, among others, 1H-indole, methyl anthranilate, 1,3-dihydro-2H-indol-2-one and caffeine. The major HoS headspace compounds were linalool, α-terpineol, 1,8-cineole, 1H-indole, methyl anthranilate, and cis-jasmone. Characteristic compounds from HoS extract were caffeine, 1H-indole, 1,3-dihydro-2H-indol-2-one, methyl anthranilate, and phenylacetonitrile. However, HO headspace composition was significantly different in comparison to NE and HoS with respect to phenylacetaldehyde and linalool derivatives abundance that appeared as the consequence of the hive conditions and the bee enzyme activity. C. unshiu honey traceability is determined by chemical markers: phenylacetaldehyde, phenylacetonitrile, linalool and its derivatives, as well as 1H-indole, 1,3-dihydro-2H-indol-2-one, and caffeine