Cream production and biological in vivo/in vitro activity assessment of a novel boron-based compound derived from quercetin and phenyl boronic acid.

Boronic acids constitute an important class of synthetic intermediates due to their high chemical stability, ease of use, moderate organic Lewis acid properties, reduced reactivity profiles and numerous biological activities such as antibacterial and antioxidant. The present study documents the synthesis and characterization of a novel boronic ester compound (3,5,7-trihydroxy-2- (2-phenyl benzo [d] [1,3,2] dioxaborol-5-yl) -4H-chromen-4-a) which was derived from phenyl boronic acid and quercetin. The new boron-based compound was used in the cream formulation after evaluating its antioxidant, antibacterial, anti-enzyme, anticancer activities and electrochemical oxidation behaviour. Furthermore, the cream has been dermatologically and microbiologically tested. Also, histological evaluation of the agent was estimated on multiple rat organs by hematoxylin-eosin staining method. Antioxidant potential of the new compound was tested by ABTS cation radical (IC50: 0.11 ± 0.01 µg/mL), DPPH free radical scavenging (IC50: 0.14 ± 0.01 µg/mL), and CUPRAC (A0.5: 1.73 ± 0.16 µg/mL) methods, respectively. The compound determined to have a dominant antioxidant activity. In addition, the synthesized compound had no toxic effect on the healthy cell line (PDF), while having a very high (IC50: 18.76 ± 0.62 µg/mL) cytotoxic effect on the cancerous cell line (MCF-7). In general, the compound showed moderate acetylcholinesterase enzyme activity (IC50: 115.63 ± 1.16 µg/mL), high butyrylcholinesterase (IC50: 3.12 ± 0.04 µg/mL), antiurease (IC50: 1.10 ± 0.06 µg/mL), and antithyrosinase (IC50: 11.52 ± 0.46 µg/mL) enzyme activities. In addition, the compound was found to be effective against Escherichia coli (ATCC 25922) bacteria studied at concentrations of 6.50 mg/mL. Moreover, the test results of the boronic ester compound used in the cream formulation demonstrated that it was microbiologically and dermatologically appropriate. Histologic analysis showed that the control group and experimental group were at similar properties without significant change. The phenyl boronic acid derivative compound synthesized from quercetin may have higher biological activity potential than quercetin. Due to the high biological activity potential of the synthesized compound, it has the potential to be used in food, feed, pharmaceutical and cosmetic industries.

Investigation of cytotoxic and apoptotic effects of 63 compounds obtained from Salvia species: Promising anticancer agents.

Since ancient time, Salvia L. species have been commonly used to treat colds, bronchitis, tuberculosis, heart diseases, and menstrual and digestive disorders in traditional medicine all around the world. They have been also used as tea and spice. Studies indicated that diterpenes and triterpenes isolated from Salvia species possess various pharmacological and biological effects such as anti-inflammatory, antiviral, cytotoxic, antioxidant, and hepatotoxic activities. Flavones were also shown to have antimicrobial, antioxidant, and cytotoxic potentials. Salvia extracts also exhibit anti-Alzheimer, antiseptic, cardiovascular, antihypertensive, and antituberculous effects. To investigate the effects of 63 secondary metabolites from Salvia species on cell viability and apoptosis, Salvia secondary metabolites including 25 phenolics, 4 fatty acids, 19 abietane diterpenoids, 12 triterpenoids, and three steroids were examined on healthy cell line (PDF), breast cancer (MCF-7), and colon cancer (HT-29) cell lines using MTT method. In addition, the effects of rosmarinic acid, 6,7-dehydroroyleanone, acetyl royleanone, ferruginol, carnosic acid, carnosol, cryptotanshinone, ß-sitosterol, and ursolic acid on pro-apoptotic Bax and antiapoptotic Bcl-2 protein expression levels were investigated by Western Blot method. PRACTICAL APPLICATIONS: Phenolic compounds (apigenin, chrysin, and luteolin) and diterpenes (especially dihydrotanshinone I, carnosic acid, and carnosol), and almost all of the triterpenes exhibited high toxic effects on healthy cell line. Cytotoxic effects of cryptotanshinone, 12-hydroxy abieta-1,3,5(10),8,11,13-hexaene, 12-demethylmulticauline, 6,7-dehydroroyleanone, acetyl royleanone, ferruginol, ursolic acid, and 3-acetyl lupeol were relatively higher than their toxic effects. Acetyl royleanone, 6,7-dehydroroyleanone, carnosic acid, and cryptotanshinone were found to have anticancer potential based on their modulating effects on the expression levels of Bax and Bcl-2 proteins which play important roles in the regulation of apoptosis. The results of the present study showed that acetyl royleanone, cryptotanshinone, 6,7-dehydroroyleanone, carnosic acid, and cryptotanshinone have potential to be used in the pharmaceutical industry.

Phytochemical Contents of Different Parts of the Seeded Raisins from the South-East Anatolia: Enzyme Inhibitory Potential of Pulp Extracts.

In this study, some phytochemical properties of six seeded raisin species that are mainly cultivated in Southeastern Anatolia were investigated. Additionally, some physical and quality characteristics, phenolic contents (by LC-MS/MS; Liquid Chromatography Tandem Mass/Mass Spectrometer System), anticholinesterase, and antioxidant capacities (DPPH; 2,2-diphenyl-1-picrylhydrazyl free-radical scavenging, ABTS; 2,2-azinobis(3-ethylbenzothiazoline-6-sulphonic acid cation-radical scavenging activity and CUPRAC; cupric reducing antioxidant capacity) of the cultivars were investigated on ground raisins. In all three methods, the antioxidant activity values of seed extracts were determined to be higher than those of leaf and pulp extracts. Remarkably, the seed extract of Banazi Siyahi showed the highest antioxidant activity in ABTS (IC50 : 4.35±0.02 µg/mL), DPPH (IC50 : 10.78±0.78 µg/mL), and CUPRAC (A0.5: 9.33±0.45 µg/mL) methods. Additionally, the ethanol extracts of all pulp samples showed higher anticholinesterase activity against acetyl-(AChE) and butyrylcholinesterase (BChE) enzymes than galantamine. According to the LC-MS/MS results, catechin (21.362 mg analyte/g extract) and epicatechin (44.667 mg analyte/g extract) found to be quite rich in Kerküs seed extract and isoquercitrin (116.873 mg analyte/g extract) and astragalin (31.915 mg analyte/g extract) detected to be quite rich in Banazi Siyahi leaf extract. Considering the mineral content of the varieties and the soil samples they grow in, all of the grape varieties analyzed in the study was found to be rich. Based on these findings, it might be suggested that Banazi Siyahi and Kerküs varieties have potential to be utilized in pharmaceutical and food industries, due to their contents of catechins, isoquercitrin and astragalin.

Determination of essential oil and biological activities of Hypericum ternatum Poulter and H. scabrum L. species collected from different localities: is H. scabrum an alternative to multifunctional species ST JOHN'S WORT (H. perforatum)?

The importance of Hypericum L. species, being used in traditional medicine, in the scientific world is increasing day by day. Hypericum species are plants that have been used in the treatment of many diseases for a long time and have healing properties. In the current study, the essential oil compositions of Hypericum scabrum L. and H. ternatum Poulter collected from different localities in Turkey were determined by GC-MS/FID. In addition, their antioxidant, cytotoxic, and antimicrobial activities with their enzyme inhibitory potentials (cholinesterase, urease, tyrosinase, elastase, and collagenase) were investigated. Also, in vitro and in silico studies of the major components of the species have been carried out on the studied enzymes. It was determined that four H. scabrum samples mainly contained α-pinene (55.99%-62.80%) while three H. ternatum samples contained 2-methyloctane (9.45%-22.39%) and α-pinene (12.75%-33.08%). While H. scabrum essential oils possessed significant biological activity potential except for the antimicrobial activity, H. ternatum samples did not. All essential oil samples of H. scabrum exhibited a high cytotoxic effect (IC50 21.67 ± 0.34 to 34.67 ± 0.45 µg/mL) against colon cancer cell line (HT-29) and indicated higher acetyl-(Inhibiton% 83.30 ± 1.90 to 93.08 ± 1.04) and butyryl-cholinesterase (Inhibiton% 80.58 ± 1.19 to 93.28 ± 1.99) inhibitory activity than the standard compound, galantamine. Furthermore, all samples of H. scabrum showed high tyrosinase (Inhibiton% 69.00 ± 1.64 to 95.25 ± 1.42) and elastase (Inhibiton% 27.58 ± 0.34 to 36.69 ± 0.18) inhibitory effects. These biological activity results indicated that H. scabrum essential oil could be used in the cosmetic and pharmaceutical industries.

Development and Validation of a Novel LC-MS/MS Method for the Quantitation of 19 Fingerprint Phytochemicals in Salvia Species: A Chemometric Approach.

Being traditionally utilized mainly as appetizers and herbal teas as well as used to ease abdominal pains, colds and gastrointestinal issues, the genus Salvia L. has gained significant consideration owing to its remarkable phytochemicals and industrial importance. The present study aimed to develop and validate an LC-MS/MS method for the qualitative and quantitative investigation of 19 fingerprint phytochemicals in six endemic Salvia species. The validation parameters of the developed LC-MS/MS method were repeatability (intermediate precision), recovery (accuracy), limits of detection and quantification, linearity and uncertainty (U% at 95% confidence level (k = 2)). Reversed-phase HPLC separation and mass spectrometry parameters were optimized for each analyte. Ethanol extracts of the studied Salvia species collected in three consecutive years were screened for their fingerprint phytochemicals by using the developed and validated LC-MS/MS method. Moreover, studied Salvia species were subjected to multivariate analysis such as principal component analysis techniques to demonstrate the variabilities in phytochemical contents by years and parts of the samples. Roots, flowers, leaves, branches and whole plant of the Salvia species collected in 2015, 2016 and 2017 were used for the analyses. It was observed that the roots and branches of Salvia species were similar in terms of their salvianolic acid A, caffeic acid, and 6,7-dehydroroyleanone components. Accordingly, apigenin, rosmarinic acid, luteolin 7-O-glucoside, caffeic acid, salvianolic acid B, and 6,7-dehydroroyleanone were notable phytochemicals that were present in the studied Salvia species.

Determination of fingerprints contents of different extracts and parts of six endemic Salvia taxa by GC-MS: Source species for valuable compounds with drug or drug potential.

Public use of Salvia species and their importance in the scientific world is continually increasing. It is known that this use and the importance of Salvia species are mostly due to the terpenoid compounds that they contain. In this context, the terpenoid-steroid-flavonoid contents of extracts of six endemic Salvia (S. kurdica, S. pseudeuphratica, S. rosifolia, S. siirtica, S. cerino-pruinosa var. cerino-pruinosa and S. cerino-pruinosa var. elazigensis) species prepared with different solvents were determined by gas chromatography-mass spectrometry. Within the framework of the ingredient analysis, content analysis of the ethanol extracts of the root, branch, leaf and flower parts of the species collected in the same period between 2015 and 2017 years was performed. In general, extracts prepared with chloroform and ethanol were found to contain a wide variety of compounds while petroleum ether extracts were found to contain much less varied compounds. In addition, in general, root extracts are richer in terpenoid compounds than aerial part extracts. Some species can be used as source species in terms of ferruginol, cryptanol, 6,7-dehydroroyleanone, lup-(20)29-ene-2α-hydroxy-3ß-acetate, salvigenin and ß-sitosterol contents (52,114.28, 75,979.08, 101,247.41, 40,071.29, 33,952.13 and 34,010.90 µg analyte/g extract, respectively).

Essential Oil, Aroma, and Fatty Acid Profiles of Five Endemic Salvia Taxa from Turkey with Chemometric Analysis.

The essential oil, aroma, and fatty acid compositions of Salvia cerino-pruinosa var. cerino-pruinosa, S. cerino-pruinosa var. elazigensis, S. pseudeuphratica, S. rosifolia, and S. kurdica collected in 2015, 2016, and 2017 were analysed by GC-FID, GC/MS/Head Space and GC/MS. The results were evaluated chemometrically for principal component analyses and hierarchical clustering analyses using Minitab program. The main components of their essential oils have been determined as germacrene-D (41.79 % highest for these species), camphor (55.80 %), linalool (33.38 %), caryophyllene oxide (32.99 %), 1,8-cineole (26.30 %) and geraniol acetate (31.63 %), while for aroma 1,8-cineole (45.95 % highest for these species), camphor (58.54 %), D-limonene (40.83 %), linalool (27.67 %) and cis-linalool oxide (25.87 %). Oleic (72.65 % highest for these species), erucic (44.61 %), 2-palmitoleic (42.17 %) and nervonic (32.61 %) acids were determined as major components of their fatty acids. According to the PCA and HCA, the essential oil components of these Salvia taxa unaffected by the years, and accordingly, the Salvia taxa grouped among themselves.

Cannabinoids for SARS-CoV-2 and is there evidence of their therapeutic efficacy?

To combat the coronaviruses and their novel variants, therapeutic drugs and the development of vaccines that are to be effective throughout human life are urgently needed. The endocannabinoid system (ECS) acts as a modulator in the activation of the microcirculation, immune system, and autonomic nervous system, along with controlling pharmacological functions such as emotional responses, homeostasis, motor functions, cognition, and motivation. The ECS contains endogenous cannabinoids, cannabinoid receptor (CBRs), and enzymes that regulate their biosynthesis, transport, and degradation. Moreover, phytocannabinoids and synthetic cannabinoids that mimic the action of endocannabinoids also play an essential role in the modulation of the ECS. Cannabinoids, the main constituents of cannabis (Cannabis sativa L.), are therapeutic compounds that have received international attention in the health field due to their therapeutic properties. Recently, they have been tested for the treatment of COVID-19 due to their antiviral properties. Indeed, cannabinoid-type compounds, and in particular cannabidiol (CBD), isolated from glandular trichomes found in the calyx of cannabis flowers with reported antiviral properties is hypothesized to be a therapeutic option in the ministration of SARS-CoV-2 consorted with COVID-19 disease. The relevant articles were determined from the database search published mainly in Web of Science, Google scholar, PubMed, Crossref, and ClinicalTrials.gov database during the pandemic period. The articles were evaluated for the therapeutic potentials, mechanisms of action of cannabinoids, the roles of the ECS in the immune system, impact of cannabinoids in SARS-CoV-2 septic, especially if they address the application of cannabinoids as drugs for the curability and management of SARS-CoV-2 and its novel variants. Although the evidence needed to be considered using cannabinoids in the control and treatment of viral diseases is currently in its infancy, they already offer an opportunity for clinicians due to their effects in relieving pain, improving appetite, and improving childhood epilepsy, especially in cancer and human immunodeficiency virus (HIV/AIDS) patients. In addition to these, the most recent scientific evidence emphasizes their use in the treatment of the coronavirus infected patients. In brief, all preclinic and clinic studies that have been reported show that, through the cannabinoid system, cannabinoids, particularly CBD, have many mechanisms that are effective in the treatment of patients infected by SARS-CoV-2. Thus, more extensive studies are necessary in this area to fully identify the effects of cannabinoids on SARS-CoV-2.

Phytochemical Fingerprints and Bioactivities of Ripe Disseminules (Fruit-Seeds) of Seventeen Gundelia (Kenger-Kereng Dikeni) Species from Anatolia with Chemometric Approach.

Gundelia species are known as "Kenger-kereng dikeni" in Anatolia, and their aerial parts are consumed as food. Also, roots and seeds (disseminules) of the Gundelia species are used to prepare gum and coffee. The chemical contents of ethanol and hexane extracts of disseminules of 17 Gundelia species, 13 of them are endemic, were studied using LC/MS/MS and GC/MS. Additionally, their antioxidant potential and enzyme inhibitory capacity against acetyl- and butyryl-cholinesterase, urease, and tyrosinase were determined. The unsaturated fatty acid ratios of Gundelia species were higher than their saturated fatty acid ratio. The highest sum of oleic and linoleic acid was detected in G. tournefortii var. tenuisecta (70.42 %). ß-Sitosterol, α-amyrin, 3-acetyllupeol were identified in 17 Gundelia species by GC/MS, while chlorogenic acid and luteolin by LC/MS/MS as major compounds. The ethanol and hexane extracts of G. siirtica, G. rosea, and G. mesopotamica indicated good cholinesterase inhibitory activity. Among all species, ethanol extract of G. colemerikensis exhibited the best activity in ABTS (IC50 : 32.30±0.98 µg/mL), DPPH (IC50 : 59.91±0.89 µg/mL), and CUPRAC (A0.5 : 57.41±1.03 µg/mL) assays. Ethanol extract of G. colemerikensis also displayed the highest inhibitory activity against butyrylcholinesterase (51.14±0.25 % at 200 µg/mL), urease (51.71±1.75 % at 200 µg/mL), and tyrosinase (39.50±0.85 % at 200 µg/mL) enzymes. According to the chemometric analysis of fatty acids, four groups were observed. Therefore, it is suggested that G. colemerikensis can be used in the pharmaceutical, food, and cosmetic industries due to its antioxidant and enzyme inhibition properties.

A Detailed Chemical and Biological Investigation of Twelve Allium Species from Eastern Anatolia with Chemometric Studies.

Allium species are widely consumed as food all over the world. The phenolic profile of ethanol extracts of aerial parts and roots of 12 Allium species, collected from five different Eastern Anatolia regions, were studied using LC-MS/MS. In vitro antioxidant, anticholinesterase, cytotoxic and antimicrobial activities were also tested. The multivariate analyses were performed using principal component and hierarchical cluster analyses. Seventeen of 27 standard compounds were detected in all Allium species. The major components were mainly identified as quinic acid, malic acid, vanillin, and p-coumaric acid. The aerial parts possessed better antioxidant activity than roots. Aerial parts of A. atroviolaceum, A. chrysantherum, A. kharputense, and A. shirnakiense exhibited high cytotoxic activity against DLD-1 colon cancer cell lines (IC50 12.5 µg/mL). A. shatakiense and A. vineale demonstrated good antimicrobial activity against S. aureus and E. coli (MIC 75 µg/mL). According to chemometric analysis, differences were detected between aerial parts and the roots. The aerial parts of A. atroviolaceum, A. chrysantherum, A. kharputense, and A. shirnakiense could be potent in the pharmaceutical industry while A. shatakiense and A. vineale in the food industry after further investigations.

Selective in-vitro Enzymes' Inhibitory Activities of Fingerprints Compounds of Salvia Species and Molecular Docking Simulations.

Recently Nutrition and Food Chemistry researches have been focused on plants and their products or their secondary metabolites having anti-alzheimer, anti-cancer, anti-aging, and antioxidant properties. Among these plants Salvia L. (Lamiaceae) species come into prominence with their booster effects due to high antioxidant contents, which have over 900 species in the world and 98 in Turkey. Some Salvia species are already in use as herbal treatment of vessel stiffness, Dementia like problems and cancer. Recently some species of Salvia are of extensive research topic. In this study, inhibitory potentials of secondary metabolites, rosmarinic acid, salvigenin, salvianolic acid A and B, tanshinone I and IIA, cyrtotanshinone, dihydrotanshinone I, carnosic acid, carnosol, and danshensu sodium salt were investigated against acetylcholinesterase, butyrylcholinesterase, urease and tyrosinase enzymes both in-vitro and in slico in detail. Elevated inhibitory effects on acetyl- and butyryl-cholinesterase of dihydrotanshinone I (IC50: 1.50 ± 0.02 and 0.50 ± 0.01 µg/mL, respectively), carnasol (IC50: 11.15 ± 0.05 ve 3.92 ± 0.03 µg/mL) and carnosic acid (IC50: 31.83 ± 0.65 ve 4.12±0.04 µg/mL) were observed. Furthermore, all other secondary metabolites were active against butyrylcholinesterase. Anti-urease (42.41 ± 0.85%) and anti-tyrosinase (39.82 ± 1.16%) activities of tanshinone I were also observed. Potential inhibitory effects of these molecules on target proteins were investigated using DOCK and molecular dynamics calculations. Dock score analysis and Lipinski parameters were demonstrated that these ligands are potential inhibitors against relevant enzymes. Our findings suggest that Salvia species can be utilized as a ptential source of anti-alzheimer active compounds for designing novel products.

A GC-MS method validation for quantitative investigation of some chemical markers in Salvia hypargeia Fisch. & C.A. Mey. of Turkey: Enzyme inhibitory potential of ferruginol.

In the present study, a GC-MS method used for quantitative screening of 26 compounds (sclareolide, sclareol, ferruginol, cryptanol, 6,7-dehydroroyleanone, suginal, 9,10-dihydro-7,8-dimethyl-2-(1-methylethyl) phenanthren-3-ol, sugiol, inuroyleanone, 12-demethylmulticauline, 7α-hydroxy-ß-sitosterol, stigmasterol, sitosterol, salvigenin, sinensetin, α-amyrin, lupeol, lupenone, 3-acetyl lupeol, 1α,21α-dihydroxy-2,3-(1'1'-dimethyl-dioxymethylene) urs-9(11),12-dien, uvaol, betulin, pyxinol, lup-(20),29-ene-2α-hydroxy-3ß-acetate, betulin 3ß, 28ß-diacetate, 21α-hydroxy,2α,3ß-diacetoxy urs-9(11),12-dien) specific to Turkish Salvia species was developed and validated. According to the GC-MS analysis results, Salvia hypargeia Fisch. & C.A. Mey. roots were found to be rich in ferruginol (30787.97 µg/g extract) and lupenone (23276.21 µg/g extract), and leaves in lupeol (20625.92 µg/g extract). Additionally, the essential oil and aroma contents of this species were identified by GC-MS technique. According to the LC-MS/MS results, especially S. hypargeia leaf extract was rich in rosmarinic acid (38035.7 µg/g extract) and isoquercitrin (4136.91 µg/g extract). Furthermore, anticholinesterase, antiurease, antityrosinase and antielastase inhibitory, antioxidant, cytotoxic activities of the ethanol extracts, essential oil, and major components of the species were evaluated. Antioxidant potentials of all extracts of this species were quite high in all studied antioxidant methods. Moreover, butyrylcholinesterase and elastase inhibitory capacities of ferruginol, the major component of S. hypargeia roots, were notable. For these reasons, this species has a high potential for food and pharmaceutical industries. PRACTICAL APPLICATIONS: This new GC-MS method was applied to S. hypargeia Fisch. & C.A. Mey. and it indicated that this species possessed high amount of ferruginol and lupeol, and that this species could be used for their natural sources. According to the results of the activity studies (antioxidant, anticholinesterase, tyrosinase, elastase, and cytotoxic), this method was used to exhibit which compound may be responsible for the activities. This developed and validated method could be easily applied to determine major/active/toxic secondary metabolites of Salvia species which are used and/or could be used in pharmaceutical, cosmetic, and food industries.

Selective in vitro and in silico enzymes inhibitory activities of phenolic acids and flavonoids of food plants: Relations with oxidative stress.

In this study, the inhibitory potentials of food originated 34 phenolic acids, and flavonoid compounds were screened against acetylcholinesterase, butyrylcholinesterase, urease, and tyrosinase enzymes. All compounds included in this study exhibited high antioxidant activity with an ignorable cytotoxic activity. In general, they also showed poor anti-urease and anti-tyrosinase activities. Compounds in aglycone form (quercetin, myricetin, chrysin, and luteolin) showed strong anticholinesterase activities. No relation was observed between the tested bioactivities except from the case that aglycone compounds exhibited a strong positive relationship between antioxidant activities and anticholinesterase activity. Interestingly, there was a relation between the molecular weights of aglycone compounds and their anticholinesterase activities. The study showed that flavonoids with molecular mass of 250-320 g/mol have high potential of anticholinesterase activities and are valuable for future experiments on animals and humans. Potential inhibitory effects of these molecules on target proteins were investigated using docking and molecular dynamics calculations.

A Detailed Biological and Chemical Investigation of Sixteen Achillea Species' Essential Oils via Chemometric Approach.

Representatives of the Achillea genus are widely used as foods or nutraceuticals. Considering the increasing demand for herbal dietary supplements with health promoting effects, the objective of this research was to evaluate the chemical composition and biological activities of the essential oils obtained from sixteen Achillea species (A. biebersteinii, A. wilhelmsii subsp. wilhelmsii, A. aleppica subsp. zederbaueri, A. vermicularis, A. monocephala, A. nobilis, A. goniocephala, A. sintenisii, A. coarctata, A. kotschyi subsp. kotschyi, A. millefolium subsp. millefolium, A. lycaonica, A. spinulifolia, A. teretifolia, A. setacea, and A. schischkinii). Anticholinesterase, antiurease, antityrosinase enzymes inhibition, antioxidant, antimicrobial, toxic and cytotoxic activities of obtained essential oils were investigated. DPPH activities were found to be very low in all studied samples, while ABTS and CUPRAC antioxidant activities were found to be moderate. In addition, all samples were found to have moderate anticholinesterase and antimicrobial effects. It has been determined that the studied species have low cytotoxicity and high toxicity. Besides, chemical composition of the essential oils were determined by GC/MS and the results were chemometrically analyzed. The chemometric analyses of Achillea species collected from nine different regions were accomplished by principal component analysis (PCA) and hierarchical cluster analysis (HCA) techniques. According to the PCA analysis, A. nobilis subsp. neilreichii was found to be different from all studied species in terms of essential oil composition. The major components found in these species were piperitone, camphor, α-terpinene, eucalyptol, artemisia ketone, endo-borneol, ß-eudesmol and verbenol. The fact that camphor was toxic and found in majority of the studied species stands out as a remarkable result.

Phytochemical and biological investigations on two Nepeta species: Nepeta heliotropifolia and N. congesta subsp. cryptantha.

In the present study, the essential oil and aroma compositions of Nepeta heliotropifolia (NH) and N. congesta subsp. cryptantha (NC) were determined by gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detector (GC/FID), and their phenolic compounds by liquid chromatography-tandem mass spectrometry (LC-MS/MS). In addition, antioxidant, cytotoxic, anticholinesterase, urease, and tyrosinase activities of essential oils obtained from NH and NC aerial parts and ethanol extracts prepared from different parts of NH and NC were investigated. The major constituent of water-distilled essential oils was found to be germacrene D (36.7% and 38.5%, respectively), and their main aroma component was eucalyptol (48.0% and 24.7%, respectively). Among the studied parts of NH and NC, their flowers extracts were found to be the richest in phenolic compounds and in which the most abundant compound was rosmarinic acid (8,909.91 and 4,317.20 µg/g, respectively). NH and NC flower extracts exhibited also strong antioxidant activity in DPPH, ABTS, and CUPRAC assays. Among the tested samples, NH essential oil indicated the best cytotoxic effect against PDF, HT-29, and MCF-7 (IC50 52.34, 25.89, and 44.70 µg/ml, respectively), and the highest butyrylcholinesterase (77.21 ± 1.12% inhibition) and moderate acetylcholinesterase (41.36 ± 0.69% inhibition) inhibitory activities. PRACTICAL APPLICATIONS: This is the first report on the essential oil and aroma compositions, the phenolic compounds, the antioxidant with total phenolic and flavonoid contents, cytotoxic, anticholinesterase, urease, and tyrosinase activities of Nepeta heliotropifolia and N. congesta subsp. cryptantha, except for their essential oil compositions. The reported results suggested that Nepeta heliotropifolia and N. congesta subsp. cryptantha flowers being rich in rosmarinic acid and having strong antioxidant potential, and NH essential oil possessing significant cytotoxic and butyrylcholinesterase inhibitory effect could be source for nutraceutical, food, and drug industries.

Trace Element Analysis by ICP-MS and Chemometric Approach in Some Euphorbia Species: Potential to become a Biomonitor.

In this study, the branch, leaves, flowers, roots and mixed parts of different nine Euphorbia species were analyzed for their trace element contents by using ICP-MS. The samples were digested by concentrated nitric acid and hydrogen peroxide in a microwave by ICP-MS before the analysis. The accuracy and precision of the method was evaluated by CRM 1573a Tomato Leaves. Trace element contents accumulated in different parts of each sample were contrasted. Minitab Statistical Software Inc., programme was used for the multivariate analysis of 12 toxic metals of seeds, roots, branches, leaves, flowers, and mixed parts of Euphorbia species collected from Diyarbakir, Kayseri, Malatya, Mardin, Trabzon, and Van cities.When the studied Euphorbia species are compared in terms of their metal contents; V, Tl, Cr, and Ni metals in E. eriophora, Ba in E. aleppica, As and Co metals in E. segiieriana, Ag and Se metals in E. craspedia, Cu and Cd metals in E. fistulosa, Cs and Pb metals in E. grisophylla, Zn in E. macroclada and also Rb and Sr metals in E. denticulata were determined higher. It was determined that the studied species accumulated some metals at highly amounts especially in the root and leaf parts. In general, it can be said that Euphorbia species have high potential to become a biomonitor. For this reason, it can be predicted that these species will be used as ornamental plants in landscape architecture due to both their toxic metals retention properties and their beautiful appearance.

A comprehensive LC-MS/MS method validation for the quantitative investigation of 37 fingerprint phytochemicals in Achillea species: A detailed examination of A. coarctata and A. monocephala.

The current study aims to optimize and validate a comprehensive LC-MS/MS method for the quantification of 37 phytochemicals (15 phenolic acids, 17 flavonoids, 3 non-phenolic organic acids, 1 phenolic aldehyde and 1 benzopyrene) in Achillea species. Though Achillea species were chosen as real life samples, the current method is applicable to a wide range of plant species. The developed method was fully validated in terms of linearity, accuracy (recovery), inter-day and intra-day precision (repeatability), limits of detection and quantification (LOD/LOQ) and relative standard uncertainty (U% at 95% confidence level (k = 2)). Reversed-phase ultrahigh performance liquid chromatography was optimized to achive optimum separation for 37 phytochemical compounds and to overcome the suppression effects. MS detection was performed using a triple quadrupole mass spectrometer and negative or positive ionization modes were optimized for each analyte. Multiple reaction monitoring (MRM) was used to quantify the analytes, related molecular ions and transition ions were optimized. Phytochemical screening of ethanol and methanol-chloroform extracts of root and aerial parts of A. coarctata and A. monocephala were performed by using the developed and validated LC-MS/MS method. Root and aerial parts of both species have considerable amounts of certain phenolic-nonphenolic acids (quinic, malic, fumaric, chlorogenic and vanillic acids) and flavonoids (rutin, hesperidin, isoquercitrin, apigetrin, luteolin, apigenin). Additionally, total phenolic and flavonoid amounts, antioxidant (DPPH free radical scavenging assay, ABTS radical cation decolorization assay, ß-carotene lipid peroxidation test system and CUPRAC cupper reduction capacity methods), anticholinesterase, tyrosinase, urease inhibition and cytotoxic activities (on HeLa (Human Cervical Carcinoma Cell Line) of A. coarctata and A. monocephala were also investigated. It has been determined that the studied Achillea species, that are rich in total phenolic-flavonoid and chlorogenic acid contents, have high antioxidant and cytotoxic potential at the same time. According to the results of LC-MS/MS, antioxidant and cytotoxic activity studies, after detailed chemical investigation and toxicity studies on these species, A. coarctata and A. monocephala may be promoted as promising sources of natural agents and used for the development of nutraceuticals or functional food ingredients in future.

Effect of electromagnetic fields and antioxidants on the trace element content of rat teeth.

The purpose of this study was to examine the possible effect of extremely low-frequency electromagnetic fields (ELF-EMFs), from a high-voltage source, on rat teeth in terms of changes in trace elements (TEs) and the effect of antioxidants (melatonin [MLT] and Ganoderma lucidum [GL]) in counteracting these effects. We used adult male Wistar albino rats with a mean weight of 250-300 g and divided the rats into eight groups. The groups were subjected to an ELF-EMF that was applied with a high-voltage line for 8 hours/day for 26 days (Groups I, II, and III) or 52 days (Groups V, VI, and VII). Groups IV and VIII were the 26- and 52-day control/sham groups, respectively. Groups II and VI were treated with GL, and Groups III and VII were treated with MLT. MLT and GL were administered daily based on the weight of the animals and appropriate standards. At the end of the study, the rats were euthanized, and their anterior teeth were extracted. The teeth were preserved in pure water before evaluating the major TEs. At the end of the study, TE concentrations (in mg/kg) were assessed in the control and test groups. Compared with Group V, statistically significant differences in the concentrations of zinc (Zn) and strontium (Sr) were found for Group VII (ELF-EMF + MLT) (P<0.05). Therefore, ELF-EMF exposure can change the content of certain TEs in teeth and, after administering MLT and GL, the values of some of the TEs return to normal.

UHPLC-ESI-MS/MS and GC-MS Analyses on Phenolic, Fatty Acid and Essential Oil of Verbascum pinetorum with Antioxidant, Anticholinesterase, Antimicrobial and DNA Damage Protection Effects.

This paper is the first phytochemical and ABTS cation radical decolorisation activity, cupric reducing antioxidant capacity, anticholinesterase and DNA damage protection effect of endemic Verbascum pinetorum (Boiss.) O. Kuntze. Phenolic profile of V. pinetorum were qualified and quantified by UHPLC-ESI-MS/MS analysis. Malic acid (47250.61±2504.28 µg/g) and luteolin (7651.96±527.98 µg/g) were found as most abundant compounds for metanol and acetone extracts, respectively. Fatty acid and essential oil compositions were determined by GC-MS analysis. The main components of fatty acid were found to be palmitic (27.1%) and stearic (22.1%) acids. The main compounds of the essential oil were cineole (16.9%) and α-selinene (16.4%). The acetone extract was found to be more active than BHT used as a standard in ß-carotene-linoleic acid test system. In DPPH free radical scavenging activity, the acetone and methanol extracts showed higher activity than BHT at all tested concentrations. The acetone, methanol and water extracts showed strong inhibition while the acetone extract showed better activity than BHT and α-tocopherol which were used as standards in ABTS cation radical scavenging and cupric reducing antioxidant capacity assays, respectively. All extracts were found to be inactive in antialzheimer activity. The acetone extract exhibited moderate antimicrobial activity against C. albicans. The methanol extract of V. pinetorum were found no significant effect on DNA cleavage protection.