A Comparative Study of the Chemical Properties and Antibacterial Activity of Four Different Ozonated Oils for Veterinary Purposes.

Infectious skin diseases are quite common in veterinary medicine. These diseases can be caused by both bacteria and pathogenic fungi. Antimicrobial drugs are usually used for treatment. An alternative to these drugs could be ozonated oils with antibacterial and antifungal properties. Four different ozonated oils (linseed, hemp seed, sunflower, and olive) were tested in order to develop an optimal pharmaceutical form for the treatment of skin infections in animals. Chemical parameters such as acid and acidity value, iodine and peroxide value, viscosity, and infrared spectres were analysed. The ozonation of oils resulted in changes in their chemical composition. The antimicrobial activity of the tested oils was evaluated by determining the minimum inhibitory concentrations and zones of inhibition in agar. After ozonation, the acid content increased in all the tested oils. The highest acidity was found in linseed oil (13.00 ± 0.11 mg KOH/g; 6.1%). Hemp oil, whose acidity was also significant (second only to linseed oil), was the least acidified by ozonation (11.45 ± 0.09 mg KOH/g; 5.75%). After ozonation, the iodine value in oils was significantly reduced (45-93%), and the highest amounts of iodine value remained in linseed (47.50 ± 11.94 g Iodine/100 g oil) and hemp (44.77 ± 1.41 Iodine/100 g oil) oils. The highest number of peroxides after the ozonation of oils was found in sunflower oil (382 ± 9.8 meqO2/kg). It was found that ozonated hemp and linseed oils do not solidify and remain in liquid form when the temperature drops. The results showed a tendency for the reference strains of S. aureus, E. faecalis, and E. coli to have broader zones of inhibition (p < 0.001) than clinical strains. Overall, ozonated linseed oil had the highest antibacterial activity, and ozonated olive oil had the lowest, as determined by both methods. It was found that ozonated linseed oil was the most effective on bacteria, while the most sensitive were S. aureus ATCC 25923, MRSA, and S. pseudointermedius (MIC 13.5 mg/mL, 4.6 mg/mL, and 13.5 mg/mL, respectively, and sterile zones 20.67 ± 0.98 mm, 20.25 ± 0.45 mm, and 18.25 ± 0.45 mm, respectively). The aim and new aspect of this work is the characterisation of selected ozonated vegetable oils, especially hemp oil, according to chemical and antibacterial parameters, in order to select suitable candidates for preclinical and clinical animal studies in the treatment of bacterial or fungal skin infections in terms of safety and efficacy.

Investigation of Bioactive Complexes of Chitosan and Green Coffee Bean or Artichoke Extracts.

The formation of water-insoluble complexes between chitosan (ChS) and caffeoylquinic acid (CQ) derivatives present in artichoke (AE) and green coffee bean (GCBE) extracts was investigated by the equilibrium adsorption method. The UPLC/HPLC analysis revealed that the phenolic compounds accounted for 8.1% and 74.6% of AE and GCBE respectively, and CQ derivatives were the predominant compounds. According to the applied Langmuir adsorption model, anionic compounds present in natural extracts were adsorbed onto the active centers of ChS, i.e., primary amino groups. The driving forces of adsorption were electrostatic interactions between cationic groups of ChS and anionic compounds of natural extracts. Chromatographic analysis revealed that not only CQ derivatives, but also other phenolic compounds of natural extracts were attached to ChS. The release of adsorbed compounds into different media as well as the bioactive properties of complexes were also studied. With the immobilization of bioactives onto ChS, increased and prolonged ABTS•+ radical scavenging activity and decreased antifungal activity against Fusarium graminearum and Botrytis cinerea were observed compared to those of ChS. The findings of the current study highlight that the adsorption approach could be used to successfully prepare water-insoluble complexes of ChS and components of natural extracts with prolonged antioxidant activity.

Optimization of Delivery and Bioavailability of Encapsulated Caffeic Acid.

Caffeic acid is a widely distributed phenolic acid. It is described in the scientific literature that caffeic acid has poor solubility. The aim of this study was to improve the solubility of caffeic acid for better dissolution kinetics when administered orally. During the study, oral capsules of different compositions were modeled. The results of the disintegration test revealed that the excipients affected the disintegration time of the capsules. The excipient hypromellose prolonged the disintegration time and dissolution time of caffeic acid. The dissolution kinetics of caffeic acid from capsules depend on the chosen excipients. P407 was more effective compared to other excipients and positively affected the dissolution kinetics of caffeic acid compared to other excipients. When the capsule contained 25 mg of ß-cyclodextrin, 85% of the caffeic acid was released after 60 min. When the capsule contained 25-50 mg poloxamer 407, more than 85.0% of the caffeic acid was released from capsules after 30 min. The research results showed that in order to improve the dissolution kinetics of caffeic acid, one of the important steps is to improve its solubility.

Citrus × paradisi L. Fruit Waste: The Impact of Eco-Friendly Extraction Techniques on the Phytochemical and Antioxidant Potential.

Citrus fruits have been the subject of extensive research over the years due to their impressive antioxidant properties, the health benefits of flavanones, and their potential use in the prevention and treatment of chronic diseases. Grapefruit have been shown in studies to improve overall health, with numerous potential benefits, including improved heart health, reduced risk of certain cancers, improved digestive health, and improved immune system function. The development of cyclodextrin complexes is an exciting approach to increasing the content of flavanones such as naringin and naringenin in the extraction medium while improving the profile of beneficial phenolic compounds and the antioxidant profile. This research aims to optimize the extraction conditions of the flavanones naringin and naringenin with additional compounds to increase their yield from different parts of grapefruit (Citrus × paradisi L.) fruits, such as albedo and segmental membranes. In addition, the total content of phenolic compounds, flavonoids, and the antioxidant activity of ethanolic extracts produced conventionally and with -cyclodextrin was examined and compared. In addition, antioxidant activity was measured using the radical scavenging activity assay (ABTS), radical scavenging activity assay (DPPH), and ferric reducing antioxidant power (FRAP) methods. The yield of naringin increased from 10.53 ± 0.52 mg/g to 45.56 ± 5.06 mg/g to 51.11 ± 7.63 mg/g of the segmental membrane when cyclodextrins (α, ß-CD) were used; naringenin increased from 65.85 ± 10.96 µg/g to 91.19 ± 15.19 µg/g of the segmental membrane when cyclodextrins (α, ß-CD) were used. Furthermore, the results showed that cyclodextrin-assisted extraction had a significant impact in significantly increasing the yield of flavanones from grapefruit. In addition, the process was more efficient and less expensive, resulting in higher yields of flavanones with a lower concentration of ethanol and effort. This shows that cyclodextrin-assisted extraction is an excellent method for extracting valuable compounds from grapefruit.

Anti-viral and Anti-inflammatory Isoflavonoids from Ukrainian Iris aphylla Rhizomes: Structure-Activity Relationship Coupled with ChemGPS-NP Analysis.

Dried Iris rhizomes have been used in Chinese and European traditional medicine for the treatment of various diseases such as bacterial infections, cancer, and inflammation, as well as for being astringent, laxative, and diuretic agents. Eighteen phenolic compounds including some rare secondary metabolites, such as irisolidone, kikkalidone, irigenin, irisolone, germanaism B, kaempferol, and xanthone mangiferin, were isolated for the first time from Iris aphylla rhizomes. The hydroethanolic Iris aphylla extract and some of its isolated constituents showed protective effects against influenza H1N1 and enterovirus D68 and anti-inflammatory activity in human neutrophils. The promising anti-influenza effect of apigenin (13: , almost 100% inhibition at 50 µM), kaempferol (14: , 92%), and quercetin (15: , 48%) were further confirmed by neuraminidase inhibitory assay. Irisolidone (1: , almost 100% inhibition at 50 µM), kikkalidone (5: , 93%), and kaempferol (14: , 83%) showed promising anti-enterovirus D68 activity in vitro. The identified compounds were plotted using ChemGPS-NP to correlate the observed activity of the isolated phenolic compounds with the in-house database of anti-influenza and anti-enterovirus agents. Our results indicated that the hydroethanolic Iris aphylla extract and Iris phenolics hold the potential to be developed for the management of seasonal pandemics of influenza and enterovirus infections.

Fractionation and Characterization of Triterpenoids from Vaccinium vitis-idaea L. Cuticular Waxes and Their Potential as Anticancer Agents.

Fruit and leaf cuticular waxes are valuable source materials for the isolation of triterpenoids that can be applied as natural antioxidants and anticancer agents. The present study aimed at the semi-preparative fractionation of triterpenoids from cuticular wax extracts of Vaccinium vitis-idaea L. (lingonberry) leaves and fruits and the evaluation of their cytotoxic potential. Qualitative and quantitative characterization of obtained extracts and triterpenoid fractions was performed using HPLC-PDA method, followed by complementary analysis by GC-MS. For each fraction, cytotoxic activities towards the human colon adenocarcinoma cell line (HT-29), malignant melanoma cell line (IGR39), clear renal carcinoma cell line (CaKi-1), and normal endothelial cells (EC) were determined using MTT assay. Furthermore, the effect of the most promising samples on cancer spheroid growth and viability was examined. This study allowed us to confirm that particular triterpenoid mixtures from lingonberry waxes may possess stronger cytotoxic activities than crude unpurified extracts. Fractions containing triterpenoid acids plus fernenol, complexes of oleanolic:ursolic acids, and erythrodiol:uvaol were found to be the most potent therapeutic candidates in the management of cancer diseases. The specificity of cuticular wax extracts of lingonberry leaves and fruits, leading to different purity and anticancer potential of obtained counterpart fractions, was also enclosed. These findings contribute to the profitable utilization of lingonberry cuticular waxes and provide considerable insights into the anticancer effects of particular triterpenoids and pharmacological interactions.

Trends in Phenolic Profiles of Achillea millefolium from Different Geographical Gradients.

The traditional widely used raw material of Achillea millefolium is currently mainly derived from wild populations, leading to diversification and uncertainty in its quality. The aim of the study was to determine the accumulation differences of phenolic compounds between geographically distant populations of Achillea millefolium from northern and southern gradients. Plant material was collected from Gaziantep and Nevsehir provinces in Turkey and from wild populations in Lithuania. A complex of nine hydroxycinnamic acids and eleven flavonoids was identified and quantified in the methanolic extracts of inflorescences, leaves, and stems using the HPLC-PDA method. Caffeoylquinic acids predominated in leaves, while inflorescences tended to prevail in flavonoids. The PCA score plot model represented the quantitative distribution pattern of phenolic compounds along a geographical gradient of populations. The content of phenolic compounds in plant materials from northern latitudes was more than twice that of plants from southern latitudes. A significant correlation of individual phenolic compounds with latitude/longitude corresponded to their differences between two countries. Differences in accumulation of caffeoylquinic acids and flavonoids revealed several intraspecific groups within A. millefolium. Our findings suggest that spatial geographical data on the distribution of phenolic compounds in A. millefolium populations could be used as a tool to find potential collection sites for high-quality raw materials.

Phenological Variations in the Content of Polyphenols and Triterpenoids in Epilobium angustifolium Herb Originating from Ukraine.

The composition of secondary metabolites undergoes significant changes in plants depending on the growth phase and the influence of environmental factors. Therefore, it is important to determine the harvesting time of plant material for the optimum secondary metabolite profile and therapeutic activity of the primary material. The shoots of Epilobium angustifolium are used as a healing tea due to the presence of polyphenolic compounds. The aim of this study was to assess the composition of phenolic compounds and triterpenoid saponins in E. angustifolium leaves and flowers and to estimate the dynamics of their content depending on the flowering phase. Qualitative and quantitative characterisation of polyphenols and triterpenoids in E. angustifolium samples from Ukraine of three flowering phases were performed using the high-performance liquid chromatography photo diode array (HPLC-PDA) method. During the present study, 13 polyphenolic compounds and seven triterpenoids were identified in the plant material. It was noted that the largest content and the best polyphenol profile was in late flowering. The most important polyphenolic compounds in the plant material were chlorogenic acid, hyperoside, isoquercitin, and oenothein B. The triterpenoid profile was at its maximum during mass flowering, with corosolic and ursolic acids being the dominant metabolites. The results of the analysis revealed that the quantity of many of the tested metabolites in the raw material of E. angustifolium is dependent on the plant organ and flowering phase. The largest content of most metabolites in the leaves was in late flowering. In the flowers, the quantity of the metabolites studied was more variable, but decreased during mass flowering and increased significantly again in late flowering. The results show that E. angustifolium raw material is a potential source of oenothein B and triterpenoids.

Correction: Stanciauskaite et al. Balsam Poplar Buds: Extraction of Potential Phenolic Compounds with Polyethylene Glycol Aqueous Solution, Thermal Sterilization of Extracts and Challenges to Their Application in Topical Ocular Formulations. Antioxidants 2022, 11, 1771.

In the original publication [...].

Balsam Poplar Buds: Extraction of Potential Phenolic Compounds with Polyethylene Glycol Aqueous Solution, Thermal Sterilization of Extracts and Challenges to Their Application in Topical Ocular Formulations.

Phenolic compounds of natural origin have been valued for their beneficial effects on health since ancient times. During our study, we performed the extraction of phenolic compounds from balsam poplar buds using different concentrations of aqueous polyethylene glycol 400 solvents (10-30% PEG400). The aqueous 30% PEG400 extract showed the best phenolic yield. The stability of the extract during autoclave sterilization was evaluated. The extract remained stable under heat sterilization. Ophthalmic formulations are formed using different concentrations (8-15%) of poloxamer 407 (P407) together with hydroxypropyl methylcellulose (0.3%), sodium carboxymethyl cellulose (0.3%) or hyaluronic acid (0.1%). Physicochemical parameters of the formulations remained significantly unchanged after sterilization. Formulations based on 12% P407 exhibited properties characteristic of in situ gels, the gelation point of the formulations was close to the temperature of the cornea. After evaluating the amount of released compounds, it was found that, as the concentration of polymers increases, the amount of released compounds decreases. Formulations based on 15% P407 released the least biologically active compounds. Sterilized formulations remained stable for 30 days.

Effective Isolation of Picrocrocin and Crocins from Saffron: From HPTLC to Working Standard Obtaining.

Saffron is widely cultivated and used as a spice. Recently published data on the chemical composition and pharmacological potential of saffron determine its use in pharmacy and medicine. The proposed high-performance thin-layer chromatography (HPTLC) method allows good separation of 11 analytes. The saffron quality (Iran, Ukraine, Spain, Morocco samples) assessment was based on the European Pharmacopoeia monograph and ISO 3632. The HPTLC method for the safranal, crocin, and picrocrocin quantification was proposed and validated. The crocins content in Ukrainian saffron was from 17.80% to 33.25%. Based on qualitative and quantitative assessment results, the saffron sample from Zaporizhzhia (Ukraine) had the highest compounds content and was chosen to obtain the working standards of picrocrocin and crocins (trans-4GG, trans-2G, trans-3Gg) by preparative chromatography. The compounds were isolated from lyophilized extract of saffron using a Symmetry Prep C18 column (300 × 19 mm × 7 µm), and identified by spectroscopic techniques (HPLC-DAD, UPLC-ESI-MS/MS). The purity of crocins and picrocrocin was more than 97%. A novel method proposed to obtain working standards is simple and reproducible for the routine analysis of saffron quality control.

Optimization of Naringin and Naringenin Extraction from Citrus × paradisi L. Using Hydrolysis and Excipients as Adsorbent.

While flavanones exist in a variety of chemical forms, their favorable health effects are most prominent in their free form-aglycones. Their concentrations in grapefruit (Citrus × paradisi L.) extracts vary according to the extraction and hydrolysis methods used. The primary aim of this work was to maximize the yields of naringin and naringenin from various parts of fresh grapefruit fruits (flavedo, albedo, and segmental) using different extraction and hydrolysis methods. In addition, we aimed to evaluate the excipient-magnesium aluminometasilicate-and determine its influence on the qualitative composition of grapefruit extracts. Extracts were obtained by heat reflux extraction (HRE), ultrasound-assisted extraction with an ultrasonic homogenizer (UAE*), and ultrasound-assisted extraction with a bath (UAE). Ultrasound-assisted extraction using a bath (UAE) was modulated using acidic, thermal, and alkaline hydrolysis. The highest yield of naringin 8A (17.45 ± 0.872 mg/g) was obtained from an albedo sample under optimal conditions using ultrasound-assisted extraction; a high yield of naringenin 23-SHR (35.80 ± 1.79 µg/g) was produced using the heat reflux method from the segmental part. Meanwhile, ultrasonic combined with thermal hydrolysis significantly increased flavanone extraction from the albedo and segmental parts: naringin from sample 9-A (from 17.45 ± 0.872 mg/g to 25.05 ± 1.25 mg/g) and naringenin from sample 15-S (from 0 to 4.21 ± 0.55 µg/g). Additionally, magnesium aluminometasilicate demonstrated significant increases of naringenin from all treated grapefruit parts. To our knowledge, this is the first report of magnesium aluminometasilicate used as an adsorbent in flavanone extractions.

Distribution Patterns of Essential Oil Terpenes in Native and Invasive Solidago Species and Their Comparative Assessment.

The importance of invasive Solidago L. species to the environment creates a new approach to controlling their spread through the use of potentially high value raw materials. The aim of this study was to assess the distribution patterns of volatile compounds in the four Solidago spp., by identifying common and species-specific compounds with their potentials, and to confirm the origin of the spontaneous hybrid Solidago × niederederi on the basis of comparative assessment of essential oil (EO) profiles. Plant material in the flowering phase was collected in mixed populations from six different sites. The EOs were isolated separately from the leaf and the inflorescence samples by hydrodistillation for 3 h. The chemical analysis was performed by gas chromatography-mass spectrometry. Multivariate data analysis was employed to explain the interspecies relationships among Solidago spp. The results revealed the similarity among Solidago spp. EO profiles, which were dominated by monoterpenes and oxygenated compound fractions. Solidago spp. differed in species distinctive terpenes and their distribution between accessions and plant parts. Volatile compound patterns confirmed the origin of Solidago × niederederi between Solidago canadensis and Solidago virgaurea, with the higher contribution of alien species than native ones. Correct taxonomic identification of species is highly essential for the targeted collection of raw material from the wild for different applications. Solidago spp. can be considered to be underutilized sources of bioactive secondary metabolites.

Ischemia In Vivo Induces Cardiolipin Oxidation in Rat Kidney Mitochondria.

Cardiolipin is a mitochondrial phospholipid that plays a significant role in mitochondrial bioenergetics. Cardiolipin is oxidized under conditions like oxidative stress that occurs during ischemia/reperfusion; however, it is known that even during ischemia, many reactive oxygen species are generated. Our aim was to analyze the effect of in vivo ischemia on cardiolipin oxidation. Adult male Wistar rats were anesthetized; then, their abdomens were opened, and microvascular clips were placed on renal arteries for 30, 40 or 60 min, causing ischemia. After ischemia, kidneys were harvested, mitochondria were isolated, and lipids were extracted for chromatographic and mass spectrometric analysis of tetralinoleoyl cardiolipin and its oxidation products. Chromatographic and mass spectrometric analysis revealed a 47%, 68% and 74% decrease in tetralinoleoyl cardiolipin after 30 min, 40 min and 60 min of renal ischemia, respectively (p < 0.05). Eight different cardiolipin oxidation products with up to eight additional oxygens were identified in rat kidney mitochondria. A total of 40 min of ischemia caused an average of a 6.9-fold increase in all oxidized cardiolipin forms. We present evidence that renal ischemia in vivo alone induces tetralinoleoyl cardiolipin oxidation and depletion in rat kidney mitochondria.

Bioactive constituents of Iris hybrida (Iridaceae): Processing effect.

Iris genus plants are a valuable source of bioactive compounds, which are an important component for pharmaceutical development. The present article shows the potential for mineral nutrition with application of magnesium sulfate, iron chelates and potassium oxide affecting the phenolic compound contents in Iris hybrida 'Tsikavynka', I. hybrida 'Tambo' and I. hybridа 'Widecombe Fire'. The effect of mineral processing was specific to plant organs and varied in the component composition. The Iris rhizomes had an increased total phenolic compound content after treatment (up to 10% of the total isoflavonoid content, up to 8% of phenolic acids, up to 5% of γ-pyrones and up to 13% of flavonoids), determined using UV-vis spectroscopy. A positive effect of nutrition on the biosynthesis and content of individual isoflavonoids (tectoridin, nigricin d-glucoside, genistin, iristectorigenin B, nigricin, irigenin and irisolidone) and xanthone mangiferin in Iris rhizomes by HPLC was established. In addition, an increase in the chlorogenic acid amount in Iris leaves was noted. The results demonstrate the sensitivity of Iris phenylpropanoid metabolism to mineral nutrition and can be used to predict medical plant cultivation with increased content of bioactive constituents.

Innovative GACP Approaches for Obtaining the Quality Iris hybrida Leaves for the Pharmaceutical Industry.

The ever-growing demand for active compounds of plant origin contributes to the implementation of cultivation methods for medicinal plants, according to the WHO guideline "Good Agricultural and Collection Practice (GACP) for Starting Materials of Herbal Origin" to obtain high-quality raw material with the stable phytocomponent composition. Therefore, the development of the cultivation and processing stages of Iris varieties leaves is necessary and promising. The present article showed the potential of proper cultivation with GACP recommendations on affecting the phenolic compounds content in Iris×hybrida hort. 'Indian Pow Waw', Iris×hybrida hort. 'Galleon Gold', and Iris×hybrida hort. 'Mini Dinamo' leaves. The cultivation process was carried out on the experimental sites of the flowering and ornamental plants department of M.M. Hryshko National Botanical Garden of the National Academy of Sciences of Ukraine (Kyiv, Ukraine) during 2018-2021. A positive effect of Iris samples proper cultivation and content of isoflavonoids (tectoridin, nigricin D-glucoside, genistin, iristectorigenin B, nigricin, irigenin, irisolidone), xanthone mangiferin, and also chlorogenic acid in Irises leaves by HPLC has been established. According to the analysis mangiferin (7.57∼28.75 µg/g), genistin (3324.82∼14642.10 µg/g), irisolidone (673.53∼2015.81 µg/g), and irigenin (3904.37∼1595.94 µg/g) were the dominant components and these compounds can be proposed as chemical markers for Iris raw material. The obtained results indicate a significant positive effect of the introduction and observance of the proper cultivation of medicinal plants to obtain a stable bioactive compounds content, in this case, on the example of Iris genus plants. Further work on the implementation of the good practice recommendation is planned to be carried out for various medicinal plants, since only controlled cultivation makes it possible to obtain high-quality raw materials with a standardized composition.

The Effect of Traditional and Cyclodextrin-Assisted Extraction Methods on Trifolium pratense L. (Red Clover) Extracts Antioxidant Potential.

Red clover is the subject of numerous studies because of its antioxidant properties, the positive influence of isoflavones on the health, and its potential use in the prevention and treatment of chronic diseases. The right excipients, such as cyclodextrins, can increase the profile of valuable phenolic compounds in extraction media to obtain rich in antioxidants, extracts that can be used in nutraceuticals production. The aim of this study was to investigate and compare the total phenolic content, flavonoid content, and antioxidant activity of red clover aerial parts, aqueous and ethanolic extracts prepared using traditional and cyclodextrins-assisted methods. The antioxidant activity of the extracts was established using ABTS, DPPH, FRAP, and ABTS-post column methods. It was determined that cyclodextrins significantly increased total phenolic content (compared with control)-using ß-cyclodextrin 20.29% (in aqueous samples); γ-cyclodextrin 22.26% (in ethanolic samples). All the samples prepared with excipients demonstrated a strong relationship between total phenolic content and DPPH assay. Study showed that for extraction with water, the highest amounts of phenolic compounds, flavonoids and antioxidant activity will be achieved with ß-cyclodextrin, but extractions with ethanol will give the best results with γ-cyclodextrin. Therefore, cyclodextrins are a great and safe tool for obtaining rich, red clover flower extracts that are high in antioxidant activity, which can be used in the pharmaceutical industry for nutraceuticals production.

Distribution of Phenolic Compounds and Antioxidant Activity in Plant Parts and Populations of Seven Underutilized Wild Achillea Species.

Evaluation of phytochemical composition of underutilized Achillea species provides the primary selection of germplasms with the desired quality of raw material for their further applications. The aim of the study was to evaluate the comprehensive distribution patterns of phenolic compounds in seven wild Achillea spp. and their plant parts, and to assess their antioxidant activity. Plant material was collected from different sites in Turkey. A complex of hydroxycinnamic acids, flavonols and flavones was identified and quantified in methanolic extracts using HPLC-PDA method. Antioxidant activity was assessed by radical scavenging assay. The results showed that qualitative and qualitative profiles of caffeoylquinic acids and flavonoids were species-specific, explaining the characteristic patterns of their variation in the corresponding species and plant parts. The highest total amount of caffeoylquinic acids was detected in A. setacea. A. arabica exposed the highest accumulation of mono-caffeoylquinic acids and flavonoids with the greatest levels of quercetin and luteolin derivatives and the flavonol santin. Santin was detected in all plant parts of A. cappadocica, A. setacea, A. santolinoides subsp. wilhelmsii, and A. arabica. A notable antiradical capacity was confirmed in A. arabica, A. setacea and A. cappadocica plant extracts. The leaves of all studied species were found to have priority over inflorescences and stems in terms of radical scavenging activity. The new data complemented the information that may be relevant for the continuation of chemophenetic studies in the heterogeneous genus Achillea.

Comparative analysis of apocarotenoids and phenolic constituents of Crocus sativus stigmas from 11 countries: Ecological impact.

The chemical compositions of 15 saffron samples from 11 countries (Morocco, India, Italy, Spain, Germany, Switzerland, Iran, Lithuania, Ukraine, Australia, and Azerbaijan) were evaluated. The samples were analyzed regarding the impact of environmental factors on the composition of apocarotenoids and phenolic constituents. Quantification of saffron metabolites was carried out using high-performance liquid chromatography. It was found that the high content of chlorogenic acid (0.2 mg/g, Ukraine) and ferulic acid (0.28 mg/g, India) was controlled by the duration of solar radiation during plant development. The accumulation of caffeic acid (the higher content 4.88 mg/g, Ukraine) in stigmas depended on the average air temperature. In contrast, the total crocins content according to the correlation analysis depended on the duration of solar radiation, the solar UV index, and the soil type. Rutin was found in all samples (0.83-8.74 mg/g). The highest amount of crocins (average 382.45 mg/g) accumulated in saffron from Italy and Ukraine. Crocins, picrocrocin, safranal, and rutin can further serve as saffron quality markers. All validation parameters were satisfactory and high-performance liquid chromatography methods could be successfully applied for the composition assessment of saffron metabolites. Saffron extracts showed the highest antibacterial activity against Bacillus subtilis, Staphylococcus aureus, and Escherichia coli (MICs 62.5-125 µg/ml).

Effective and simple approach for colchicine determination in saffron parts.

Crocus sativus (Iridaceae) or saffron is important for its flavoring properties in the food industry and its medicinal properties in the pharmaceutical industry. Real saffron must meet the quality requirements set to FDA or ISO specifications, and in addition, must be safe and non-toxic. An "autumn crocus" or scientific name Colchicum spp. (Colchicaceae) is a plant that contains a highly toxic alkaloid colchicine. The problem is that Colchicum autumnale is often mistaken for C. sativus. This study aim is to develop the high-performance liquid chromatography method for determining the presence of colchicine in C. sativus stigma, flowers and corms, and then for solving the issue of saffron possible falsification and toxicity. The chromatographic separation was performed on ACE C18 column (5.0 µm, 250 mm × 4.6 mm), by using 0.1% acetic acid in water and acetonitrile as the mobile phase and UV detector (350 nm). Current analysis showed that all Crocus raw materials didn't contain colchicine. This confirms the safety of saffron and its parts for using in the development of substances based on them. The proposed method was successfully applied on C. autumnale corms and showed good separation and colchicine identification. The presence of tricyclic proto-alkaloid colchicine in plant products is not a new phenomenon, however, the need for new sensitive analytical methods that can detect it in food and medicine still exists. The identification and confirmation of colchicine absence in saffron raw materials are important for further herbal drug development and for the food industry.