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    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
    (Elsevier Science Bv, 2018) Yilmaz, Mustafa Abdullah; Ertas, Abdulselam; Yener, Ismail; Akdeniz, Mehmet; Cakir, Oguz; Altun, Muhammed; Demirtas, Ibrahim
    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, beta-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. (C) 2018 Elsevier B.V. All rights reserved.
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    LC/MS-MS Analyses and in vitro anticancer activity of Tourneuxia variifolia extracts
    (Taylor & Francis Ltd, 2022) Zerrouki, Sara; Mezhoud, Samia; Yilmaz, Mustafa Abdullah; Yaglioglu, Ayse Sahin; Bakir, Derya; Demirtas, Ibrahim; Mekkiou, Ratiba
    Several Saharan plants, despite their abundance of natural compounds, have received little attention. In this study, the chemical composition of polar extracts of Tourneuxia variifolia Coss. (Asteraceae), an endemic species to Algerian Sahara, was investigated and their anticancer activity was evaluated in vitro. The phytoconstituents of both ethyl acetate (EtOAc) and n-butanol (n-BuOH) extracts were screened using LC/MS-MS technique. The anticancer activity of the above extracts was measured against human cervical adenocarcinoma (HeLa) cell line. The LC/MS-MS analyses results revealed that twenty-seven phytochemicals in EtOAc extract and twenty-three in n-BuOH extract were identified and quantified from which isoquercetin and astragalin were the most present. Moreover; the EtOAc extract was found to have a strong anticancer activity (IC50: 46.797 +/- 0.060 mu g/mL). These findings identified T. variifolia as a potential plant exhibiting anticancer properties.
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    Phytochemical analysis, and antioxidant, anti-hemolytic and genoprotective effects of Quercus ilex L. and Pinus halepensis Mill. methanolic extracts
    (Journal Pharmacy & Pharmacognosy Research-Jppres, 2019) Meziti, Hicham; Bouriche, Hamama; Kada, Seoussen; Demirtas, Ibrahim; Kizil, Murat; Senator, Abderrahmane
    Context: Quercus ilex and Pinus halepensis are largely used in Algerian folk medicine as a remedy for different health problems. Aims: To determine the phytochemical composition as well as to evaluate the antioxidant, anti-hemolytic and genoprotective effects of Quercus ilex root bark extract (QIE) and Pinus halepensis young ovulate cones extract (PHE). Methods: Methanolic extracts were prepared by maceration. Phenolic compounds and flavonoids were quantified spectrophotometrically and identified by HPLC analysis. In vitro antioxidant activity of the extracts was assessed by determining the DPPH and ABTS radicals scavenging activities and ferric reducing/antioxidant power (FRAP). Protective effect of the extracts against AAPH-induced erythrocyte hemolysis was assessed. pBluescript M13 (+) plasmid DNA was used as oxidation target to evaluate DNA protective effect of the extracts. Results: Considerable phenolic compounds and flavonoids contents were found in the studied extracts. Catechin and phenolic acids (4-hydoxybenzoic, caffeic, coumaric, ferulic and gentisic acids) were identified in QIE, while catechin and cinnamic acid were identified in PHE. Both extracts scavenge DPPH radical with IC50 values of 5.67 mu g/mL and 18.87 mu g/mL, respectively. Antioxidant capacity against ABTS radical was 4.13 and 1.15 mM Trolox equivalent/mg extract. The two extracts also showed a considerable ferric reducing ability with 1.98 and 8.55 mM FeSO4/mg extract, respectively. On the other hand, both extracts exerted a significant protective effect against AAPH-induced erythrocyte hemolysis and DNA protective activities were noticed. Conclusions: Quercus ilex and Pinus halepensis extracts are important sources of bioactive compounds possessing important antioxidants, antihemolytic and genoprotective effects.
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    Protective activity of Hertia cheirifolia extracts against DNA damage, lipid peroxidation and protein oxidation
    (Taylor & Francis Ltd, 2017) Kada, Seoussen; Bouriche, Hamama; Senator, Abderrahmane; Demirtas, Ibrahim; Ozen, Tevfik; Toptanci, Bircan Ceken; Kizil, Goksel
    Context: Hertia cheirifolia L. (Asteraceae), a perennial shrub widely distributed in Northern Africa, is traditionally used to treat inflammatory disorders. Objective: The protective effect of methanol (Met E) and aqueous (Aq E) extracts of Hertia cheirifolia against DNA, lipid and protein oxidation was investigated. Materials and methods: Different concentrations (50-1000 mu g/mL) of Hertia cheirifolia aerial part extracts were examined against DNA, lipid and protein oxidation induced by H2O2 +UV, FeSO4, and Fe3+/H2O2-ascorbic acid, respectively. The DPPH center dot, metal ion chelating, reducing power and beta-carotene bleaching tests were conducted. Results: Both extracts were rich in polyphenols, flavonoids and tannins, and were able to scavenge DPPH center dot with IC50 values of 138 and 197 mu g/mL, respectively. At 300 mu g/mL, Aq E exerted stronger chelating effect (99%) than Met E (69%). However, Met E reducing power (IC50 = 61 mu g/mL) was more than that of Aq E (IC50 = 193 mu g/mL). Both extracts protected from beta-carotene bleaching by 74% and 94%, respectively, and inhibited linoleic acid peroxidation. The inhibitory activity of Aq E extract (64%) was twice more than that of Met E (32%). Interestingly, both extracts protected DNA against the cleavage by about 96-98%. At 1 mg/mL, Met E and Aq E restored protein band intensity by 94-99%. Conclusions: Hertia cheirifolia exhibits potent antioxidant activity and protects biomolecules against oxidative damage; hence, it may serve as potential source of natural antioxidant for pharmaceutical applications and food preservation. This is the first report on the protective activity of this plant against biomolecule oxidation.