Mustafa Abdullah Yilmaz

Comparative Studies on Phenolic Composition, Antioxidant, Wound Healing and Cytotoxic Activities of Selected Achillea L. Species Growing in Turkey.

Turkey is one of the most important centers of diversity for the genus Achillea L. in the world. Keeping in mind the immense medicinal importance of phenols, in this study, three species growing in Turkey, A. coarctata Poir. (AC), A. kotschyi Boiss. subsp. kotschyi (AK) and A. lycaonica Boiss. & Heldr. (AL) were evaluated for their phenolic compositions, total phenolic contents (TPC), antioxidant properties, wound healing potencies on NIH-3T3 fibroblasts and cytotoxic effects on MCF-7 human breast cancer cells. Comprehensive LC-MS/MS analysis revealed that AK was distinctively rich in chlorogenic acid, hyperoside, apigenin, hesperidin, rutin, kaempferol and luteolin (2890.6, 987.3, 797.0, 422.5, 188.1, 159.4 and 121.2 µg analyte/g extract, respectively). The findings exhibited a strong correlation between TPC and both free radical scavenging activity and total antioxidant capacity (TAC). Among studied species, the highest TPC (148.00 mg GAE/g extract) and TAC (2.080 UAE), the strongest radical scavenging (EC50 = 32.63 μg/mL), the most prominent wound healing and most abundant cytotoxic activities were observed with AK. The results suggested that AK is a valuable source of flavonoids and chlorogenic acid with important antioxidant, wound healing and cytotoxic activities. These findings warrant further studies to assess the potential of AK as a bioactive source that could be exploited in pharmaceutical, cosmetics and food industries.

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 β-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 profile by LC–MS/MS, GC/MS and antioxidant activities of the essential oils and crude extracts of two Euphorbia species

In this study, it was aimed to investigate the chemical composition and antioxidant activities of two Euphorbia species. The major component of the fatty acid compositions obtained from the petroleum ether extracts was identified as palmitic acid for Euphorbia gaillardotii and Euphorbia macroclada. The main constituents of the essential oils were identified as arachidic acid for E. gaillardotii and tetratetracontane for E. macroclada. Among the 27 studied compounds, hesperidin, rutin, hyperoside and quinic, malic, gallic and tannic acids were found to be the most abundant compounds in the two Euphorbia species. The methanol extracts of E. gaillardotii and E. macroclada showed strong antioxidant activity in all tested methods. Particularly, IC50 values of E. macroclada methanol extract that was the richest in terms of total phenolic-flavonoid contents were found to be lower than α-tocopherol and butylated hydroxytoluene in β-carotene bleaching, 2,2-diphenyl-1-picrylhydrazyl free and ABTS cation radical scavenging methods.

In vitro and in silico perspectives on biological and phytochemical profile of three halophyte species—A source of innovative phytopharmaceuticals from nature

BACKGROUND Halophytes are considered as valuable sources of traditional drugs in different countries. PURPOSE The present study aimed to evaluate biological and chemical fingerprints of three halophytes (Arthrocnemum macrostachyum (Moric.) C, Koch, Halimione portulacoides (L.) Aellen and Salicornia europaea L.). MATERIALS AND METHODS The antioxidant and enzymatic inhibitory potential (acetylcholinesterase, butyrylcholinesterase, α-amylase, α-glucosidase, and tyrosinase) were assessed. The total phenolic, flavonoid contents, and the chemical profiles were appraised using the ultra-high performance liquid chromatography-electrospray ionization-tandem mass spectrometry. Molecular docking was conducted to provide additional insights of molecular interactions of the enzymes/phytochemicals. RESULTS Ethyl acetate extract was the most efficient extract, with A. macrostachyum being the most potent towards DPPH and ABTS radicals and phosphomolybdenum assay. Ethyl acetate extract of A. macrostachyum was also the best reducing agent (CUPRAC and FRAP assays). Methanol and ethyl acetate extract of A. macrostachyum, H. portulacoides, and S. europaea showed significant enzyme inhibition potential. Ethyl acetate extract of A. macrostachyum showed the highest total phenolic (29.54 ± 0.78 mgGAEs/g extract) while the ethyl acetate extract of S. europaea was more abundant in flavonoids (18.26 ± 0.11 mgREs/g extract). Phytochemical profiling allowed the identification of several components in the methanolic extracts (16 in A. macrostachyum, 14 in H. portulacoides, and 11 in S. europaea), with quinic acid, p-coumaric acid, and rhamnetin being most abundant. Docking studies revealed that the above compounds showed scores for the enzymes tested. CONCLUSION The three halophytes studies could be considered as potential sources of biologically-active compounds for novel phytopharmaceuticals development.

Ecology and genomics of an important crop wild relative as a prelude to agricultural innovation

Domesticated species are impacted in unintended ways during domestication and breeding. Changes in the nature and intensity of selection impart genetic drift, reduce diversity, and increase the frequency of deleterious alleles. Such outcomes constrain our ability to expand the cultivation of crops into environments that differ from those under which domestication occurred. We address this need in chickpea, an important pulse legume, by harnessing the diversity of wild crop relatives. We document an extreme domestication-related genetic bottleneck and decipher the genetic history of wild populations. We provide evidence of ancestral adaptations for seed coat color crypsis, estimate the impact of environment on genetic structure and trait values, and demonstrate variation between wild and cultivated accessions for agronomic properties. A resource of genotyped, association mapping progeny functionally links the wild and cultivated gene pools and is an essential resource chickpea for improvement, while our methods inform collection of other wild crop progenitor species.Domestication reduces genetic diversity and constrains crop improvement. Here the authors identify factors that shaped species diversity in the wild progenitors of chickpea, and produce wild introgression populations that increase diversity for breeding by ~100-fold, including traits of agronomic relevance.

Chemical Profile and Biological Activities of Two Edible Plants: Chemical Investigation and Quantitative Analysis Using Liquid Chromatography Tandem Mass Spectrometry and Gas Chromatography Mass Spectrometry

The objectives of this study were to define the phenolic and fatty acid profiles, anticholinesterase, antioxidant, antimicrobial activities, and total phenolic-flavonoid contents of Lycopsis orientalis and Tragopogon latifolius var. angustifolius which have been used as food source and food supplement in Anatolia and have never been examined before. Rosmarinic and quinic acids (21.11 and 11.46 mg g–1 extract, respectively) were found to be the most abundant constituents in L. orientalis and T. latifolius var. angustifolius among the studied 27 compounds by liquid chromatography tandem mass spectrometry. In the fatty acid compositions of L. orientalis and T. latifolius var. angustifolius that were determined by gas chromatography mass spectrometry, oleic (29.1%) and palmitic (28.7%) acids were identified as the major components, respectively. The high antioxidant activity of the methanol extract of L. orientalis shows parallelism to its rosmarinic acid content. Besides, this extract showed medium anticholinesterase activity. The results of the present study proves that the L. orientalis might also be used as a food source due to its high phenolic acid content and strong antioxidant property.

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

HIGHLIGHTSA comprehensive LC–MS/MS method validation to quantify 37 phytochemicals in plants.The developed method is applicable to all plant species as well as Achillea.The studied species might be a source for chlorogenic acid, rutin and apigenin.A. monocephala; promising species in terms of antioxidant and cytotoxic activities. ABSTRACT 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, &bgr;‐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.

Identification of phenolic compounds, antioxidant activity and anti-cancer effects of the extract obtained from the shoots of Ornithogalum narbonense L.

This study aimed to examine the anti-cancer and antioxidant properties and identify the phenolic content of methanolic extract obtained from the shoots of the Ornithogalum narbonense L. (OR) species, which is used for folk-medicine and food in the Sanliurfa region of Turkey. The antioxidant activity of the extract was investigated using total phenolics, flavonoids, ABTS and CUPRAC methods. Phenolic component analysis of the plant extract was performed by LC-MS/MS. The anti-cancer property of OR extract was investigated on human colon (DLD-1), endometrium (ECC-1) cancer cells and embryonic kidney (HEK-293) cells. Cytotoxic effects were defined with MTT, apoptotic activity with DNA fragmentation ELISA and AO/EB fluorescent staining, the genotoxic effect with the comet assay and the intracellular oxidative status with TAS and TOS methods. As a result of the study, it was determined that OR extract showed an antioxidant effect, and as a result of the content analysis made with LC-MS/MS, phenolic components were determined, the most abundant being cosmosiin, followed by cinnamic acid, p-coumaric acid and quinic acid. OR extract showed cytotoxic activity on DLD-1 and ECC-1 cancer cells, while the cytotoxic effect on HEK-293 cells was determined to be low. It was determined that through OR extract, by increasing the intracellular amount of free radicals on cancer cells, led to DNA damage, which consequently led to apoptosis of the cancer cells.

Yenilebilir Dört Bitki Türünün Fitokimyasal İçeriğinin Miktarsal Tayini için LC-MS/MS Metot Validasyonu

Bitkilerde 8 fitokimyasal bilesigin (sinarin, kafeik asit, sirinjik asit, p-kumarik asit, o-kumarik asit, vanilik asit, ferulik asit, klorojenik asit) miktarsal tayini icin kapsamli bir LC-MS/MS metodu gelistirildi ve validasyon calismalari yapildi. Gelistirilen analitik metot lineerite, gerceklik (geri kazanim), gun ici ve gunlerarasi kesinlik (tekrarlanabilirlik ve tekrar uretilebilirlik), tespit ve tayin limitleri (LOD/LOQ) ve bagil standart belirsizlik (% 95 guven araliginda (k=2)) gibi parametreleri icerek sekilde tam validasyon calismalari yapildi. Kromatografik ayrim ters faz UHPLC sistemi ile yapildi. Kutle dedeksiyonu ise her analit icin negative veya pozitif iyonlasma modlari optimize edilerek uclu kuadrupol kutle spektrometresi ile gerceklestirildi. Analitlerin miktarsal tayini icin coklu reaksiyon goruntuleme (MRM) kullanildi, molekuler iyonlar ve ilgili gecis iyonlari ise optimize edildi. Metot validasyonu sonrasi, enginar (Cynara scolymus L.), brokoli (Brassica Oleracea var. Italica), karnabahar (Brassica Oleracea var. Botrytis) ve kenger gibi (Gundelia Tournefortii) bazi yenilebilir bitkilerin methanol ekstrelerinin fitokimyasal icerikleri gelistirilen ve valide edilen LC-MS/MS metodu ile tespit edildi. Analiz edilen bitkiler arasinda enginar, acik ara farkla fenolik yonunden en zengini olarak belirlendi. Ek olarak, analiz edilen tum bitkilerde klorojenik asit en bol bulunan bilesen olarak goruldu. Calisilan ornekler analiz icin secilmis bitkiler olsa da, gelistirilen LC-MS/MS metodu bitki dunyasindaki pekcok bitkiye uygulanabilirdir.

The evaluation of antioxidant and anticancer effects of Lepidium Sativum Subsp Spinescens L. methanol extract on cancer cells

In recent years, there is an increased research interest for plants which are natural sources of antioxidants. Lepidium sativum Subsp spinescens L., commonly found in South West Asia, is a plant known as a healthy nutritional source containing bio-molecules that carry anti-hypertensive, hypoglycemic, anti-asthmatic, antispasmodic, hepato-protective, chemoprotective, anti-inflammatory and anti-oxidant effects. In this study, we aimed to investigate the antioxidant content and activity of Lepidium sativum Subsp spinescens L. methanol extract on cancer cells. Methanol extract of dried Lepidium sativum Subsp spinescens L. was prepared. Total amount of phenolic compounds was determined by Slinkard and Singleton method using Folin-Ciocalteu reagent. Total flavonoid amount was determined according to Zhishen method. Antioxidant activity of the extract was evaluated by CUPRAC and ABTS radical scavenging activity assays. Cytotoxic effects of the plant extract on colon and endometrium cancer cells, and human peripheral lymphocyte cells were investigated in vitro by MTT and neutral red assays. Furthermore, the plant extract was investigated for necrotic effects by LDH assay; apoptotic activity by DNA ladder fragmentation, ELISA and acridine orange/ethidium bromide staining; and genotoxic effect by comet assay methods. Methanol extract of Lepidium sativum Subsp spinescens L. was found to have a high content of phenolic and flavonoid compounds. The extract showed significant antioxidant activity and also cytotoxic activity on colon and endometrium cancer cells in a concentration-dependent manner. Apoptotic activity and genotoxic effects were significantly increased, especially with 200 μg/ml concentrations at 48 hours incubation. In conclusion, it was determined that the extract evaluated in this study could be a natural source of antioxidants. Further molecular studies explaining chemo-preventive and chemotherapeutic effects on cancer cells are required to support anticancer efficacy of the plant.