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Of the six juniper species found in the Bulgarian flora, three of the species have controversial taxonomic positions. Juniperus pygmaea K. Koch and J. sibirica Burgsd. exhibit similar morphological characteristics to J. communis L. in terms of leaves and female cones (galbuli). This is one of the reasons why, in the recent taxonomic developments, J. pygmaea and J. sibirica were united in a common variety of J. communis , namely, J. communis var. saxatilis . However, such a grouping of species in the Flora of Bulgaria has not been adopted. This study aimed to evaluate the degrees of similarity or difference in the structure of the leaves, galbuli, seeds, and pollen of J. communis , J. sibirica , and J. pygmaea using the methods of comparative anatomy by light microscope (LM) and scanning electron microscopy (SEM) observations and complex morphological measurements. The working hypothesis of this study was that the three species would show a different degree of similarity with each other, which would clarify their taxonomic rank. The morphological parameters revealed differences between the length/width ratio of galbuli and seed length of the three species, while leaf characteristics (length and width) showed a stronger resemblance between J. sibirica and J. pygmaea . Furthermore, a greater distinction between the leaves and galbili of J. communis and J. sibirica was found. The SEM analyses showed variations in the seed shape and spermoderm among the three species. The shape of J. communis seeds was oval and elongated, while J. pygmaea seeds were pear-shaped, and J. sibirica seeds were triangular-rhombic. The length and height of striations were diverse on seed spermoderm in the three species. The epicuticular waxes of leaves, located on the tips of the anticlinal walls of the elongated epidermal cells in J. pygmaea and J. communis , were oval, while they formed raised comb-like crystals in J. sibirica . The morphological, anatomical, and SEM analysis affirmed the accepted taxonomic status of J. communis and J. sibirica as independent species within the Bulgarian flora. Based on most of the analyzed parameters, J. pygmaea exhibits significant similarity with J. sibirica . Additionally, the similar habitats of these two species support the determination of J. pygmaea as a variety or form of J. sibirica rather than J. communis ( J. sibirica forma pygmaea ).

Hippeastrum papilio (Amaryllidaceae) is a promising new source of galanthamine - an alkaloid used for the cognitive treatment of Alzheimer's disease. The biosynthesis and accumulation of alkaloids are tissue - and organ-specific. In the present study, histochemical localization of alkaloids in H. papilio's plant organs with Dragendorff's reagent, revealed their presence in all studied samples. Alkaloids were observed in vascular bundles, vacuoles, and intracellular spaces, while in other plant tissues and structures depended on the plant organ. The leaf parenchyma and the vascular bundles were indicated as alkaloid-rich structures which together with the high proportion of alkaloids in the phloem sap (49.3% of the Total Ion Current - TIC, measured by GC-MS) indicates the green tissues as a possible site of galanthamine biosynthesis. The bulbs and roots showed higher alkaloid content compared to the leaf parts. The highest alkaloid content was found in the inner bulb part. GC-MS metabolite profiling of H. papilio's root, bulb, and leaves revealed about 82 metabolites (>0.01% of TIC) in the apolar, polar, and phenolic acid fractions, including organic acids, fatty acids, sterols, sugars, amino acids, free phenolic acids, and conjugated phenolic acids. The most of organic and fatty acids were in the peak part of the root, while the outermost leaf was enriched with sterols. The outer and middle parts of the bulb had the highest amount of saccharides, while the peak part of the middle leaf had most of the amino acids, free and conjugated phenolic acids., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier GmbH. All rights reserved.)

The aims of this study were to (1) establish the intrapopulation and seasonal variation of Juniperus excelsа essential oil (EO); (2) compare the J. excelsa concrete and resinoid composition with its EO composition; and (3) investigate the potential herbicidal activity of J. excelsa EO against seeds of Papaver rhoeas L., Consolida orientalis (J.Gay) Schrödinger, Anthemis arvensis L., Avena fatua L., and Agrostemma githago L. Four independent studies were performed to meet these objectives. Twenty-eight individual trees were analyzed from two populations to establish intrapopulation and interpopulation variability of EOs yield and composition. In the seasonal dynamic study of leaf EO, samples from the same three trees and in the same population were collected in January, March, May, July, October, and December and their EO yield and composition determined. The EOs (intrapopulation and seasonal) were extracted by hydrodistillation, while the EO for the herbicidal test was obtained by steam distillation in a semi-commercial (SCom) apparatus. Overall, the EO yield varied significantly from 0.93% to 2.57%. α-Pinene (8.85-35.94%), limonene (11.81-50.08%), and cedrol (3.41-34.29%) were the predominant EO compounds in all samples (intrapopulation variability); however, trans-2,4-decadienol and β-caryophyllene were predominant in some individual trees. Four chemical groups were identified in the samples collected from two natural populations (intrapopulation). This is the first report on the compositions of J. excelsa concrete and resinoid. Cedrol (15.39%), 7-hydroxy-4-methyl-coumarin (17.63%), 1-octacosanol (36.85%), tritriacontane (16.08%), and tiacontanoic acid were the main compounds in the concrete and resinoid. Juniperus excelsa EO suppressed seed germination and seedling growth of P. rhoeas, C. orientalis, A. arvensis, A. fatua, and A. githago, demonstrating its potential to be used for the development of new biopesticides. The highest EO yield with high content of limonene and cedrol was obtained from samples harvested during the winter months (December, January, and March)., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Semerdjieva et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The cuticle is important in the interaction between the plant and its environment, especially in the dry areas. Four species of junipers from the section Sabina wild growing in the Balkans were selected to study leaf wax composition using GC/MS and GC-FID and its surface morphology under SEM to understand the correlation between the distribution and/or habitat of these species and their cuticles. SEM micrographs showed continuous, smooth epicuticular layers with crusts in all species but with a species-specific distribution of different 3D crystalloid types and different cuticle thickness. n-C33 alkane was the most abundant compound, followed by n-C29, n-C31, and n-C35, depending on the species and the site. The average chain length (N) was the lowest in J. phoenicea, but with the greatest dispersion around it. At the same time, the two most continental species (J. foetidissima and J. excelsa) show the N with the lowest dispersion around it. The statistical analyses confirmed the significance of climate on the evolution of the specific epicuticular wax composition in studied junipers., (© 2023 Wiley-VHCA AG, Zurich, Switzerland.)

Pinus heldreichii Christ. (Bosnian pine), a Tertiary relict and Balkan sub-endemic, has not been comprehensively studied for its essential oil (EO) profile and bioactivity of its different plant parts. This study aimed to determine the EO yield, composition and antimicrobial activity from different parts of P. heldreichii at three different populations (mountains) in Bulgaria. Furthermore, the study assessed the antioxidant activities of plant tissue, including leaves (needles), twigs wood, male and female cones. The EOs yield from different plant parts ranged from 0.09 % (leaves) to 0.74 % (wood of twigs), with monoterpenes being the predominant class. Limonene, α -pinene, β- caryophyllene, germacrene D, β- pinene, and β -myrcene were detected in the EO extracted from all analyzed trees. However, these compounds were not found in the EO extracted from all plant parts of the same trees. Four chemical groups (chemotypes) were identified for EO from twigs, and three chemotypes were identified for EO from leaves. The chemotypes were based on the percent ratio of the main EO constituents (>5 %). Leaves tissue showed the highest values in terms of polyphenols and flavonoids, as well as higher ABTS radical scavenging activity, while the highest antimicrobial activity against Staphylococcus aureus subsp. aureus was seen in the EOs obtained from twigs. This is the first study to identify several chemotypes based on leaf and twigs EO of P. heldreichii distributed in Bulgarian flora. Furthermore, the EO of twigs tips (TT), male cones (MC), and wood of one-two-year-old twigs (WT) of the same trees were reported for the first time. The total polyphenol, flavonoid content, and radical scavenging activity of tissues of annual twigs wood and biennial twigs wood, leaf tissue, MC tissue, and the twigs tips tissue is also reported for the first time in the accessible literature. These findings highlight the potential of P. heldreichii to provide EOs with varying compositions and bioactivities, making them suitable for nutraceutical, pharmacological, and potentially food additive applications. Furthermore, the identification of chemotype accessions in this study suggests their selection for the development of new forest crop as a source for natural products with desirable composition and bioactivity., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Satureja pilosa Velen., S. coerulea Janka and S. kitaibelii Wierzb. ex Heuff. are Balkan endemic species, and they are distributed in restricted territories, primarily found in dry grasslands, rocky slopes, and stony habitats. This study presents the results of the first embryological and micromorphological analyses of three Satureja species ( S. pilosa , S. kitaibelii , and S. coerulea ) from the Bulgarian flora. The aim of this study was to establish the features of the male and female reproductive sphere, as well as surface characteristics of leaves, stem, and calyx in order to understand the mode of reproduction, character, size and state of species populations and delimitation. For the embryological study, flowers and flower buds in different developmental stages were collected from plants of natural populations and treated with the classic paraffin method. Reproductive capacity was assessed using the following approaches: (1) acetocarmine test for pollen viability, (2) tetrazolium test (TTZ) for seed viability, and (3) germination test. The surfaces characteristics of leaves, stem, and calyx were analyzed by scanning electron microscopy (SEM). As a result, the study revealed the structures of the male (M) and female (F) generative spheres as well as the processes of gamete development, pollination, and endosperm and embryo formation. It was found that the three Satureja species exhibited a high pollen viability but low seed viability and germination. The SEM analysis showed both common and distinct micromorphology features regarding epidermis, calyx and stem surfaces among the three Satureja species. Notably, the S. coerulea surfaces (leaves, calyx, stem) were clearly distinguishable from the other two Satureja species. Regarding the nutlet surfaces, it was observed that the exocarp surfaces of S. coerulea and S. kitaibelii have a reticulate convex type surface and comprise two types of cells: (1) highly bulging, bubble-like cells; and (2) flat cells with numerous striations. On the other hand, the exocarp of S. pilosa displayed rectangular or polygonal shapes without bubble-like cells, and it had a tabular to slightly convex type surface. Additionally, nutlets (seeds) of both S. coerulea and S. pilosa exhibited distinct papilla formations resembling non-glandular trichomes seen on the ends of the nutlets for S. coerulea and over the entire surface for S. pilosa . The characteristics of the embryological structures and processes, along with the absence of apomixis, characterized the three studied Satureja species as sexually reproducing. The established balanced processes and stable structures contribute to their high reproductive potential and population stability. However, these traits may also decrease their adaptability to environmental changes.

Hypericum species and especially H. perforatum L. are well known for their therapeutic applications. The present study assessed the essential oil (EO) composition, and antifungal and aphid suppression activity of seven Bulgarian Hypericum species. The EOs were analyzed by GC-MS-FID. Two experiments were conducted. In the first experiment, H. perforatum , H. maculatum , and H. hirsutum were used. Additionally, the EO composition of H. perforatum extracted via hydrodistillation (ClevA) and via commercial steam distillation (Com) were compared. The second experiment compared the EOs of H. perforatum , H. cerastoides , H. rumeliacum , H. montbretii , and H. calycinum (flowers and leaves) extracted via hydrodistillation and collected with n -hexane. Overall, the EO constituents belonged to four classes, namely alkanes, monoterpenes, sesquiterpenes, and fatty acids. The main class for compounds in H. maculatum and H. perforatum (section Hypericum ) were sesquiterpenes for both experiments except for H. perforatum (Com). Hypericum montbretii (section Drosocarpium ) EO had monoterpenes (38.09%) and sesquiterpenes (37.09%) as major groups, while H. hirsutum EO (section Taeniocarpium ) contained predominately alkanes (67.19%). Hypericum hirsutum EO contained cedrol (5.04%), found for the first time in Hypericum species. Fatty acids were the main compounds in H. cerastoides (section Campylopus ), while monoterpenes were the most abundant class in H. rumeliacum and H. calycinum EOs. α -Pinene and germacrene D were the major EO constituents of all analyzed Hypericum species except for H. hirsutum and H. cerastoides. Hypericum perforatum EO (Com) had significant repellent and insecticidal activity against two aphid species, Rhopalosiphum padi (Bird Cherry-oat aphid) and Sitobion avenae (English grain aphid) at concentrations of 0%, 1%, 2.5%, 3.5%, 4.5%, and 5%. The tested EOs did not show significant activity against selected economically important agricultural fungal pathogens Fusarium spp., Botrytis cinerea , Colletotrichum spp., Rhizoctonia solani , and Aspergillus sp. The EO of the Hypericum species found in the Bulgarian flora could be utilized for the development of new biopesticides for aphid control.

Alkanna tinctoria (L.) Tausch Boraginaceae is a medicinal plant whose root is used for its antimicrobial and anti-inflammatory properties. A. tinctoria roots have been subject to numerous studies. However, the aerial parts have been explored less. The objective of the present study was to compare the chemical profile of aerial parts and roots as well as the total alkannin content in roots of 11 populations of the species from different floristic regions of Bulgaria. Methanolic extracts from 22 samples were analyzed by GC/MS. Phenolic, fatty, and organic acids, sterols, polyols, fatty alcohols, and sugars were identified. Ononitol (4-O-methyl-myo-inositol) was found as the main compound in the aerial parts. The total alkannin content in the roots was evaluated by the spectrophotometric method and compared with that of the commercial product. Populations with high alkannin content and rich in other bioactive compounds were identified. A relatively low genetic diversity in the studied populations was observed. The present study is the first comprehensive study on metabolite profiles and genetic diversity of the Bulgarian populations of A. tinctoria . The occurrence of ononitol in the aerial parts of the species is reported for the first time, as well as the phenolic acid profiles of the species in both aerial parts and roots. The results showed that aerial parts of the plant are also promising for use as a source of valuable biologically active substances.

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

This study compared the essential oils (EO) composition of Helichrysum arenarium (Bulgarian populations) with that of the cultivated species H. italicum . The EO composition of H. arenarium and H. italicum were analyzed via gas chromatography. In general, 75 components were identified in H. arenarium EO and 79 in H. italicum EO. The predominant constituents in H. arenarium EO were α -pinene (34.64-44.35%) and sabinene (10.63-11.1%), which affirmed the examined population as a new chemical type. Overall, the main EO constituents of H. italicum originating in France, Bosnia and Corsica were neryl acetate (4.04-14.87%) and β -himachalene (9.9-10.99%). However, the EOs profile of H. italicum introduced from the above three countries differed to some extent. D-limonene (5.23%), italicene, α -guaiene and neryl acetate (14.87%) predominated in the H. italicum introduced from France, while α -pinene (13.74%), δ-cadinene (5.51%), α -cadinene (3.3%), β -caryophyllene (3.65%) and α -calacorene (1.63%) predominated in plants introduced from Bosnia. The EOs of the plants introduced from France and Corsica had similar chemical composition and antimicrobiological activity.

The objective of this study was to assess the biological activity of essential oils (EOs) of four Juniperus species obtained via two different distillation methods and their potential as biopesticides. The studied factors were juniper species ( Juniperus communis L., J. oxycedrus L., J. pygmaea C. Koch., and J. sibirica Burgsd), plant sex (male (M) and female (F)), and distillation method (hydrodistillation via a standard Clevenger apparatus (ClevA) and semi-commercial (SCom) steam distillation). The hypothesis was that the EO will have differential antioxidant, antimicrobial, and insecticidal activities as a function of plant species, plant sex, and distillation method. The two distillation methods resulted in similar EO composition within a given species. However, there were differences in the EO content (yield) due to the sex of the plant, and also differences in the proportions of some EO components. The concentration of α-pinene, β-caryophyllene, δ-cadinene and δ-cadinol was dissimilar between the EO of M and F plants within all four species. Additionally, M and F plants of J. pygmaea , and J. sibirica had significantly different concentrations of sabinene within the respective species. The EOs obtained via ClevA extraction showed higher antioxidant capacity within a species compared with those from SCom extraction. All of the tested EOs had significant repellent and insecticidal activity against the two aphid species Rhopalosiphum padi (bird cherry-oat aphid) and Sitobion avenae (English grain aphid) at concentrations of the EO in the solution of 1%, 2.5%, and 5%. The tested EOs demonstrated moderate activity against selected pathogens Fusarium spp., Botrytis cinerea , Colletotrichum spp., Rhizoctonia solani and Cylindrocarpon pauciseptatum . The results demonstrate that the standard ClevA would provide comparable EO content and composition in comparison with SCom steam distillation; however, even slight differences in the EO composition may translate into differential bioactivity.

Juniperus excelsa M. Bieb and J. sabina L. contain essential oil (EO), while J. sabina also contains podophyllotoxin, which is used as a precursor for anti-cancer drugs. Two studies were conducted. The first assessed the variability in the EO profile and podophyllotoxin concentration of the two junipers, depending on the location and tree gender. The main EO constituents of J. excelsa were α-cedrol, α-limonene and α-pinene, while the constituents in J. sabina were sabinene, terpinen-4-ol, myrtenyl acetate and α-cadinol. The podophyllotoxin yield of 18 J. sabina accessions was 0.07-0.32% ( w/w ), but this was not found in any of the J. excelsa accessions. The second study assessed the effect of hydrodistillation (Clevenger apparatus) and steam distillation (in a semi-commercial apparatus) on the EO profile and bioactivity. The extraction type did not significantly alter the EO composition. The EO profiles of the two junipers and their accessions were different and may be of interest to the industry utilizing juniper leaf EO. Breeding and selection programs could be developed with the two junipers (protected species) in order to identify chemotypes with (1) a high EO content and desirable composition, and (2) a high concentration of podophyllotoxin in J. sabina . Such chemotypes could be established as agricultural crops for the commercial production of podophyllotoxin and EO.

Grecian juniper ( Juniperus excelsa M. Bieb.) is an evergreen tree and a rare plant found in very few locations in southern Bulgaria. The aim of this study was to evaluate the phytochemical content and antioxidant potential of J. excelsa unripe and ripe galbuli from three different locations in Bulgaria. The essential oil content ranged between 1.9% and 5.1%, while the lipid fraction yield was between 4.5% and 9.1%. The content of total chlorophyll was 185.4-273.4 μg/g dw. The total carotenoid content ranged between 41.7 and 50.4 μg/g dw of ripe galbuli, and protein content was between 13.6% and 16.4%. Histidine (5.5 and 8.0 mg/g content range) and lysine (4.0 and 6.1 mg/g) were the major essential amino acids. The antioxidant potential of the 95% and 70% ethanol extracts was analyzed using four different methods. A positive correlation between the antioxidant potential and phenolic content of the galbuli was found. The results obtained in this study demonstrated the differences in phytochemical composition and antioxidant capacity of J. excelsa galbuli as a function of maturity stage and collection locality.

The Balkans endemic species Alkanna primuliflora Griseb., A. stribrnyi Velen., and A. graeca Boiss. & Spruner have limited distribution in the Balkan Peninsula and a large variation in the morphological characteristics. The populations of the three Alkanna species in the Bulgarian flora are small and fragmented. There are no previous reports on the chemical profile or on the embryology of these species. The hypothesis was that the limited distribution of A. primuliflora, A. stribrnyi, and A. graeca was due to their reproductive capacity and genetic diversity. Furthermore, we hypothesized that the three species will contain pyrrolizidine alkaloids (PAs), as other species of the genus Alkanna (Boraginaceae), but they would have differential alkaloids composition. The population genetic structure and differentiation showed a clear distinction between species and revealed average levels of genetic diversity among the natural populations of the three Alkanna species. The embryological investigation observed stability of the processes in the male and female generative spheres and high viability of mature pollen and embryo of the three species. The normal formation of male and female gametophytes without deviations or degenerative processes, and observed levels of genetic diversity between Alkanna individuals are important in maintaining the size and resilience of the Alkanna populations. Eight alkaloids were identified by GC-MS in A. primuliflora and A. graeca and six alkaloids in A. stribrnyi. The main pyrrolizidine alkaloids (PAs) in all investigated species was triangularine. A. primuliflora and A. graeca showed similar chemical composition that comprised 9-angeloylretronecine, 7-tigloylretronecine, 9-tigloylretronecine, triangularicine, dihydroxytriangularine, dihydroxytriangularicine, whereas, in A. stribrnyi 9-tigloylretronecine, triangularicine and dihydroxytriangularicine were not found. This is the first report on the presence of PAs in A. primuliflora, A. stribrnyi and A. graeca. Besides, this is the first report on the embryology of these endemic species. The results contribute to the knowledge of the three endemic Alkanna species and will facilitate policy-making and defining new strategies for their conservation., Competing Interests: NO authors have competing interests.

Genus Galanthus (Amaryllidaceae) includes 19 species in Europe and the Middle East. The Flora of Bulgaria recognizes two species: G. nivalis L. and G. elwesii Hook. Galanthus elwesii is characterized by relatively high morphological variability, leading some authors to identify some populations as G. gracilis Celak. However, the occurrence of G. gracilis in the Bulgarian flora is disputed. The hypothesis was that populations previously identified as G. gracilis belong indeed to a separate species. Therefore, the objective of this study was to compare G. nivalis and G. elwesii with plants from populations previously identified as G. gracilis . Morphological, DNA, embryological and anatomical analyzes were conducted to meet the objective. The morphological characteristics and DNA dendrogram revealed that G. gracilis and G. elwesii were situated in the same cluster and had significant morphological similarity, whereas plants from populations identified as G. nivalis were dissimilar in morphology and situated in a separate cluster. The revealed features of the generative sphere showed similarities across the species. Scanning electron microscopy (SEM) analyses of the surface revealed that the anticillinal walls of G. elwesii and G. gracilis were straight, while those of G. nivalis were wavy. This research demonstrated that the plants of G. elwesii and those from populations identified as G. gracilis are morphologically, embryologically and genetically similar, thus refuting the hypothesis. This study did not provide sufficient evidence to support the claim of the existence of G. gracilis in the Bulgarian flora; the populations identified as G. gracilis in Bulgaria may be forms of G. elwesii ., (© 2019 The Author(s). Published by Elsevier Ltd.)

The chemical profile and antioxidant capacity of Juniperus virginiana , J. excelsa , and J. sabina essential oil (EO) fractions as a function of time was the subject of this study. The hypothesis was that, capturing EO in sequential timeframes during hydrodistillation would generate fractions containing unique compositions and antioxidant capacity. In J. virginiana , the highest limonene (43%) was found in the 0⁻5 min oil fraction, with safrole (37%) being highest in the 10⁻20 and 20⁻40 min fractions, and elemol (34%) being highest in the 160⁻240 min fraction. In J. excelsa , α-pinene (34-36%) was the highest in the 0⁻5 min fraction and in the control (non-stop 0⁻240 min distillation) oil, limonene (39%) was the highest in the 0⁻10 min fractions and cedrol (50-53%) was the highest in the 40⁻240 min fractions. In J. sabina , sabinene (80%) was highest in the 0⁻3 min fraction. The highest antioxidant capacity of J. virginiana was demonstrated by the 5⁻10 min fraction; the one in J. sabina by the 3⁻10 min fraction; and, the one in J. excelsa , by the control. The kinetics regression models that were developed can predict EO composition of the three juniper species eluted at different timeframes. Various industries could benefit from the results from this study.

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