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.

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.

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.

Triterpenoids and steroids isolated from Anatolian Capparis ovata and their activity on the expression of inflammatory cytokines.

CONTEXT: Capparis L. (Capparaceae) is grown worldwide. Caper has been used in traditional medicine to treat various diseases including rheumatism, kidney, liver, stomach, as well as headache and toothache. OBJECTIVE: To isolate and elucidate of the secondary metabolites of the C. ovata extracts which are responsible for their anti-inflammatory activities. MATERIALS AND METHODS: Buds, fruits, flowers, leaves and stems of C. ovata Desf. was dried, cut to pieces, then ground separately. From their dichloromethane/hexane (1:1) extracts, eight compounds were isolated and their structures were elucidated by NMR, mass spectroscopic techniques. The effects of compounds on the expression of inflammatory cytokines in SH-SY5Y cell lines were examined by qRT-PCR ranging from 4 to 96 µM. Cell viability was expressed as a percentage of the control, untreated cells. RESULTS: This is a first report on isolation of triterpenoids and steroids from C. ovata with anti-inflammatory activity. One new triterpenoid ester olean-12-en-3ß,28-diol, 3ß-pentacosanoate (1) and two new natural steroids 5α,6α-epoxycholestan-3ß-ol (5) and 5ß,6ß-epoxycholestan-3ß-ol (6) were elucidated besides known compounds; oleanolic acid (2), ursolic acid (3), ß-sitosterol (4), stigmast-5,22-dien-3ß-myristate (7) and bismethyl-octylphthalate (8). mRNA expression levels as EC10 of all the tested seven genes were decreased, particularly CXCL9 (19.36-fold), CXCL10 (8.14-fold), and TNF (18.69) by the treatment of 26 µM of compound 1 on SH-SY5Y cells. DISCUSSION AND CONCLUSIONS: Triterpenoids and steroids isolated from C. ovata were found to be moderate-strong anti-inflammatory compounds. Particularly, compounds 1 and 3 were found to be promising therapeutic agents in the treatment of inflammatory and autoimmune diseases.

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.

Anticholinesterase and Antioxidant Activities of Natural Abietane Diterpenoids with Molecular Docking Studies.

BACKGROUND: Alzheimer's Disease (AD) is one of the most prevalent causes of dementia in the world, and no drugs available that can provide a complete cure. Cholinergic neurons of the cerebral cortex of AD patients are lost due to increased activity of cholinesterase enzymes. OBJECTIVE: Acetylcholinesterase (AChE) and Butyrylcholinesterase (BuChE) are the two major classes of cholinesterases in the mammalian brain. The involvement of oxidative stress in the progression of AD is known. Thus, the objective of this study is to determine strong ChE inhibitors with anti-oxidant activity. METHODS: In this study, 41 abietane diterpenoids have been assayed for antioxidant and anticholinesterase (both for AChE and BuChE) properties in vitro, which were previously isolated from Salvia species, and structurally determined by spectroscopic methods, particularly intensive 1D- and 2DNMR and mass experiments. Molecular modeling studies were performed to rationalize the in vitro ChE inhibitory activity of several abietane diterpenoids compared with galantamine. RESULTS: Thirteen out of the tested 41 abietane diterpenoids exhibited at least 50% inhibition on either AChE or BuChE. The strongest inhibitory activity was obtained for Bractealine against BuChE (3.43 µM) and AChE (33.21 µM) while the most selective ligand was found to be Hypargenin E against BuChE enzyme (6.93 µM). A full correlation was not found between anticholinesterase and antioxidant activities. The results obtained from molecular modelling studies of Hypargenin E and Bractealine on AChE and BuChE were found to be in accordance with the in vitro anti-cholinesterase activity tests. CONCLUSION: Abietane diterpenoids are promising molecules for the treatment of mild-moderate AD.

SPE-HPLC Determination of Chlorogenic and Phenolic Acids in Coffee.

A new solid phase extraction-high-performance liquid chromatography method with ultraviolet detection was developed and validated for the determination of three chlorogenic acids (5-O-caffeoylquinic, 3-O-caffeoylquinic and 4-O-caffeoylquinic acids) and six phenolic acids (caffeic, ferulic, sinapic, protocatechuic, p-hydroxybenzoic acid (PHBA) and vanillic acid (VA)) in coffee bean samples. Extraction was performed using hydrophilic-lipophilic balance cartridges. Separations were accomplished using a C18 guard column (10 × 2.1 mm, 3 µm) and a C18 analytical column (50 × 2.1 mm, 3 µm). o-Phosphoric acid solution (0.08%) and methanol/water/acetonitrile (85:10:5) solution were used as mobile phase with a gradient system. A UV detector was used at 325 nm for 5-O-caffeoylquinic, caffeic, 3-O-caffeoylquinic, 4-O-caffeoylquinic, ferulic, sinapic acids, and 215 nm for protocatechuic, PHBA and VA. Calibration equations and coefficients of determination were determined by least-squares method with weighting factor. Limits of detection and quantification were in the range of 0.15-0.69 and 0.46-2.09 mg/L, respectively. Precision and accuracy of the proposed method were investigated with coffee sample spiked at low, medium and high concentrations. The developed and validated method was applied for the determination of nine phenolic compounds in seven coffee bean samples from different origins with high accuracy and repeatability.

Capparis ovata treatment suppresses inflammatory cytokine expression and ameliorates experimental allergic encephalomyelitis model of multiple sclerosis in C57BL/6 mice.

Since ancient times, Capparis species have been widely used in traditional medicine to treat various diseases. Our recent investigations have suggested Capparis ovata's potential anti-neuroinflammatory application for the treatment of multiple sclerosis (MS). The present study was designed to precisely determine the underlying mechanism of its anti-neuroinflammatory effect in a mouse model of MS. C. ovata water extract (COWE) was prepared using the plant's fruit, buds, and flower parts (Turkish Patent Institute, PT 2012/04,093). We immunized female C57BL/6J mice with MOG35-55/CFA. COWE was administered at a daily dose of 500mg/kg by oral gavage either from the day of immunization (T1) or at disease onset (T2) for 21days. Gene expression analysis was performed using a Mouse Multiple Sclerosis RT² Profiler PCR Array, and further determinations and validations of the identified genes were performed using qPCR. Whole-genome transcriptome profiling was analyzed using Agilent SurePrint G3 Mouse GE 8X60K microarrays. Immunohistochemical staining was applied to brain sections of the control and treated mice to examine the degree of degeneration. COWE was further fractionated and analyzed phytochemically using the Zivak Tandem Gold Triple Quadrupole LC/MS-MS system. COWE remarkably suppressed the development of EAE in T1, and the disease activity was completely inhibited. In the T2 group, the maximal score was significantly reduced compared with that of the parallel EAE group. The COWE suppression of EAE was associated with a significantly decreased expression of genes that are important in inflammatory signaling, such as TNFα, IL6, NF-κB, CCL5, CXCL9, and CXCK10. On the other hand, the expression of genes involved in myelination/remyelination was significantly increased. Immunohistochemical analysis further supported these effects, showing that the number of infiltrating immune cells was decreased in the brains of COWE-treated animals. In addition, differential expression profiling of the transcriptome revealed that COWE treatment caused the down regulation of a group of genes involved in the immune response, inflammatory response, antigen processing and presentation, B-cell-mediated immunity and innate immune response. Collectively, these results suggest anti-neuroinflammatory mechanisms by which COWE treatment delayed and suppressed the development of EAE and ameliorated the disease in mice with persistent clinical signs.

In vitro biological activities and fatty acid profiles of Pistacia terebinthus fruits and Pistacia khinjuk seeds.

This study reports in vitro anticholinesterase, antioxidant and antimicrobial effects of the n-hexane, dichloromethane, ethanol and ethanol-water extracts prepared from Pistacia terebinthus L. fruits and Pistacia khinjuk Stocks seeds as well as their total phenolic and flavonoid contents, and fatty acid compositions. Ethanol and ethanol-water extracts of both species exhibited higher anticholinesterase activity than galanthamine. Among ABTS, DPPH and CUPRAC assays, the highest antioxidant capacity of the extracts was found in the last one. P. terebinthus ethanol extract being rich in flavonoid content showed the best cupric reducing effect. All extracts possessed no antimicrobial activity. The main fatty acid in P. terebinthus fruits (52.52%) and P. khinjuk seeds (59.44%) was found to be oleic acid. Our results indicate that P. terebinthus fruits and P. khinjuk seeds could be a good source of anticholinesterase compounds, and could be phytochemically investigated.

Chemical profile and biological activities of Veronica thymoides subsp. pseudocinerea.

CONTEXT: In Turkey, Veronica species (Plantaginaceae) have been used as a diuretic and for wound healing in traditional medicine. OBJECTIVE: To examine the fatty acid and essential oil profiles, the antioxidant, anticholinesterase, antimicrobial, and DNA damage effects of Veronica thymoides P.H. Davis subsp. pseudocinerea M.A. Fischer as a potential source of natural active compounds. MATERIALS AND METHODS: GC/MS was used to analyze essential oil and fatty acid obtained from whole plant. The antioxidant activity was evaluated by the ß-carotene-linoleic acid test system, DPPH-free and ABTS cation radicals scavenging, and cupric reducing antioxidant capacity assays. The anticholinesterase and antimicrobial activities were determined by Ellman and broth macrodillution methods, respectively. The effect of the methanol extract on DNA cleavage was investigated. RESULTS: Hexatriacontene (21.0%) was found to be the main constituent in essential oil, and linoleic acid (25.2%) and palmitic acid (20.6%) in fatty acid. Methanol extract demonstrated the best IC50 values in lipid peroxidation (49.81 ± 0.31 µg/ml) and DPPH-free radical scavenging activity (15.32 ± 0.17 µg/ml). Methanol and water extracts possessed strong ABTS cation radical scavenging activity with IC50 values 9.15 ± 0.28 and 8.90 ± 0.1 µg/ml, respectively. The acetone extract exhibited moderate butyrylcholinesterase inhibitory activity. The highest antimicrobial activity was determined in methanol extract against Escherichia coli with 31.25 µg/ml MIC value. Inhibition of methanol extract on plasmid DNA cleavage by OH radicals was found to be 93.32% at 500 µg/ml. CONCLUSION: The methanol extract having strong antioxidant and DNA damage effects could be investigated phytochemically to find natural active compounds.

Essential oil compositions and anticholinesterase activities of two edible plants Tragopogon latifolius var. angustifolius and Lycopsis orientalis.

This is the first report in the literature on essential oil compositions of Tragopogon latifolius var. angustifolius and Lycopsis orientalis which were analysed by using GC-FID and GC-MS techniques. The main constituents of T. latifolius var. angustifolius were identified as α-selinene (10.5%), 2,5-di-tert octyl-p-benzoquinone (9.5%) and valencene (7.0%); however, the main components of L. orientalis were identified as heptacosane (10.5%), τ-muurolene (9.6%) and tetratetracontane (9.4%). The essential oils of T. latifolius var. angustifolius and L. orientalis species exhibited moderate inhibitory activity against acetyl- and butyryl-cholinesterase enzymes at 200 µg/mL.

Chemical compositions by using LC-MS/MS and GC-MS and biological activities of Sedum sediforme (Jacq.) Pau.

In this research, the chemical composition and biological activities of various extracts obtained from whole parts of Sedum sediforme (Jacq.) Pau were compared. The amounts of total phenolic and flavonoid components in crude extracts were determined by expression as pyrocatechol and quercetin equivalents, respectively. All of the extracts (petroleum ether, acetone, methanol, and water) obtained from S. sediforme showed strong antioxidant activity in four tested methods. Particularly, the IC50 values of the methanol extract, which was the richest in terms of total phenolic and flavonoid contents, were found to be lower than those of α-tocopherol and BHT in ß-carotene bleaching (9.78 ± 0.06 µg/mL), DPPH free radical scavenging (9.07 ± 0.07 µg/mL), and ABTS cation radical scavenging (5.87 ± 0.03 µg/mL) methods. Furthermore, the methanol extract of S. sediforme showed higher inhibition activity than galanthamine against acetyl- and butyryl-cholinesterase enzymes. Also, acetone and methanol extracts exhibited moderate antimicrobial activity against Candida albicans. The main constituents of fatty acid and essential oil were identified as palmitic acid (C16:0) (28.8%) and α-selinene (20.4%), respectively, by GC-MS. In the methanol extract of S. sediforme, quercetin, rutin, naringenin, and protocatechuic, p-coumaric, caffeic, and chlorogenic acids were detected and quantified by LC-MS/MS. Results of the current study showed that the methanol extract of S. sediforme may also be used as a food supplement.

Chemical constituents and biological activities of Cirsium leucopsis, C. sipyleum, and C. eriophorum.

Two endemic Cirsium species, C. leucopsis DC. and C. sipyleum O. Schwarz, and C. eriophorum (L.) Scop. growing in Turkey were investigated to establish their secondary metabolites, fatty acid compositions, and antioxidant and anticholinesterase potentials. Spectroscopic methods were used to elucidate the structures of thirteen known compounds (p-hydroxy-benzoic acid, vanillic acid, cis-epoxyconiferyl alcohol, syringin, balanophonin, 1'-O-methyl-balanophonin, apigenin, kaempferol-3- O-ß-D-glucopyranoside, kaempferol-3-O-α-L-rhamnopyranoside, taraxasterol, taraxasterol acetate, ß-sitosterol, ß-sitosterol-3-O-ß-D-glucopyranoside). cis-Epoxyconiferyl alcohol and 1'-O-methyl- balanophonin were isolated for the first time from Cirsium species. Palmitic acid (47.1%) was found to be the main fatty acid of C. leucopsis, linoleic acid in both C. sipyleum (42.1%) and C. eriophorum (37.8 %). Assays of ß-carotene bleaching, scavenging of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals, 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium (ABTS) cation radicals, and superoxide anion radicals, as well as cupric reducing antioxidant capacity (CUPRAC) were used to determine the antioxidant activities of the extracts and isolated compounds. Vanillic acid, balanohonin, and kaempferol-3-O-aαL-rhamnopyranoside exhibited strong antioxidant activity. Taraxa-terol was a potent inhibitor of acetyl- and butyrylcholinesterase activity, respectively.

Indole alkaloids from Vinca major and V. minor growing in Turkey.

A new indole alkaloid, 11-hydroxypolyneuridine, was isolated from Vinca major subsp. major L. and the known indole alkaloids vallesiachotamine and isovallesiachotamine from Vinca minor L. This is the first report on the alkaloids of both Vinca species growing in Turkey; vallesiachotamine and isovallesiachotamine were isolated from a Vinca species for the first time. V. minor may be considered as a new source for these two alkaloids due to their occurrence in high amount in the aerial parts of the plant. The alkaloid extracts of the two Vinca species were found to have high lipid peroxidation inhibitory and DPPH radical scavenging activities. Anticholinesterase activity of the extracts was also very strong.

Antioxidant and anticholinesterase activities of eleven edible plants.

CONTEXT: Consumers have become more interested in beneficial effects of vegetables, fruits, and tea to protect their health. OBJECTIVE: The antioxidant potential and anticholinesterase activity of eleven edible plants were investigated. MATERIAL AND METHODS: The dichloromethane, ethanol and water extracts prepared from celery [Apium graveolens L. (Umbelliferae)], Jerusalem artichoke [Helianthus tuberosus L. (Compositae)], spinach [Spinacia oleracea L. (Chenopodiaceae)], chard [Beta vulgaris L. var. cicla (Chenopodiaceae)], purslane [Portulaca oleracea L. (Portulacaceae)], ispit, or borage [Trachystemon orientale (L.) G. Don (Boraginaceae)], garden rocket [Eruca sativa Mill. (Brassicaceae)], red cabbage [Brassica oleracea L. var. capitata f. rubra DC. (Cruciferae)], lime flower [Tilia tomentosa Moench (Tiliaceae)], cinnamon [Cinnamomum cassia Presl. (Lauraceae)], and rosehip [Rosa canina L. (Rosaceae)], were tested to determine their antioxidant and anticholinesterase activities by using CUPRAC (cupric reducing antioxidant capacity) and Ellman methods, respectively, for the first time. RESULTS: As a result, the dichloromethane, ethanol and water extracts of cinnamon showed the best antioxidant effect among the extracts of the tested plants. The ethanol extract of cinnamon exhibited 63.02% inhibition against acetylcholinesterase and 85.11% inhibition against butyrylcholinesterase (BChE) at 200 µg/mL concentration while the dichloromethane extract of garden rocket possessed the highest inhibition (91.27%) against BChE among all the tested extracts. DISCUSSION AND CONCLUSION: This study indicated that the ethanol extract of cinnamon may be a new potential resource of natural antioxidant and anticholinesterase compounds.

Antioxidant and anticholinesterase constituents from the petroleum ether and chloroform extracts of Iris suaveolens.

The aim of this study was to investigate the phytochemical, antioxidant and anticholinesterase activities of Iris suaveolens. After determining total phenolic and flavonoid contents of the petroleum ether, chloroform and methanol extracts prepared from the rhizomes, the antioxidant capacity of the extracts was established using ß-carotene-linoleic acid and CUPRAC methods. The chloroform extract which was rich in phenolic content exhibited the highest inhibition of lipid peroxidation in the ß-carotene-linoleic acid system, and the best cupric reducing antioxidant capacity among the tested extracts. The petroleum ether extract indicated moderate anticholinesterase activity while the chloroform extract revealed significant butyrylcholinesterase inhibitory activity (75.03 ± 1.29%). Spectroscopic methods were used for the structural elucidation of the compounds (1-13) isolated from the petroleum ether and chloroform extracts. Coniferaldehyde (6), having the highest antioxidant activity in the ß-carotene-linoleic acid assay at 25 and 50 µg/mL, demonstrated also the best effect in the CUPRAC method among the tested compounds (1-12). 3-Hydroxyirisquinone (10) showed the best anticholinesterase activity among the tested compounds (1-4, 6-12), and coniferaldehyde exhibited almost the same butyrylcholinesterase inhibitory activity (82.60 ± 2.33%) as galantamine (86.26 ± 0.66%).

GC-MS analysis of the antioxidant active fractions of Micromeria juliana with anticholinesterase activity.

The aerial parts of Micromeria juliana (L.) Bentham ex Reichb. were extracted with light petroleum, acetone and methanol, successively. The antioxidant activity of different concentrations of the extracts was evaluated using different antioxidant tests, namely total antioxidant (lipid peroxidation inhibition activity), DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging, ferric reducing power, and metal chelating. Total antioxidant activity was determined using the beta-carotene-linoleic acid assay. Unexpectedly, the light petroleum extract exhibited strong lipid peroxidation inhibition activity. The extract was fractionated on a silica gel column and the antioxidant activity of the fractions was determined by the beta-carotene-linoleic assay at 25 microg/mL concentration. The fractions that exhibited more than 50% inhibition activity were analysed by GC and GC/MS; thus, the structure of fourteen compounds were elucidated. In addition, acetyl- and butyryl-cholinesterase inhibitory activities of the extracts were also determined in vitro. The light petroleum and acetone extracts were found to have mild butyrylcholinesterase inhibitory activity.

GC-MS analysis and antimicrobial activity of essential oil of Stachys cretica subsp. smyrnaea.

The essential oil from the aerial parts of Stachys cretica L. subsp. smyrnaea Rech. fil. (Lamiaceae), endemic to Turkey, was investigated by using GC and GC-MS. Thirty-four of 37 components, represented 99.7% of the total oil, were identified. The major components of the essential oil were trans-beta-caryophyllene (51.0%), germacrene-D (32.8%), a-humulene (3.1%), delta-cadinene (2.1%) and delta-elemene (2.1%). The antimicrobial activity of the essential oil, trans-beta-caryophyllene and five different extracts of the aerial parts of S. cretica L. subsp. smyrnaea were investigated by the standard disc diffusion method. The essential oil and trans-beta-caryophyllene exhibited antibacterial and antifungal activities. The activity increased with increasing concentrations of the essential oil and the extracts. The essential oil showed antimicrobial activity, particularly against Pseudomonas aeruginosa and Bacillus subtilis. The extracts exhibited either moderate or no activity.