Chemical characteristics and biological activity screening of Pistacia lentiscus mastic gum and leaves from Türkiye.

BACKGROUND: Mastic gum is a resin that is produced by Pistacia lentiscus. It has many traditional uses, dating from ancient times, such as the treatment of gastrointestinal disorders and as a food additive. In this study, the leaves and mastic gum of trees of different ages from Karaburun and the Cesme peninsula in Türkiye were examined chemically and biologically. Flavonoids, and phenolic and fatty acid components were evaluated by a liquid chromatography system coupled with high resolution mass spectrometry (LC-HRMS) and gas chromatography with flame ionization detection (GC-FID). Cytotoxicity was screened against several cancer and healthy cell lines using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Inducible nitric oxide synthase (iNOS) inhibition was determined on lipopolysaccharide (LPS)-induced murine macrophage cell line (RAW 264.7) cells. Antiviral activity was measured against avian coronavirus using an in ovo virucidal antiviral activity assay. RESULTS: The main phenolic constituents of the gum were found to be salicylic, rosmarinic, and caffeic acids whereas the most abundant compounds detected were flavonoids in the leaf extracts. The most abundant fatty acids in hexane extracts were palmitic and oleic acids. All gum extracts except 3-year-old gum had significant cytotoxic activity on HeLa (IC50 1.74 ± 0.03-4.76 ± 0.95) and PC-3 (0.64 ± 0.25-6.22 ± 1.40) cells. Moreover, reducing virus activity by fivefold or sixfold logarithmically between the range of 5-10 µg g-1 of 30-year-old gum extracts underscored the biological activity. CONCLUSION: In ovo antiviral activity studies on the P. lentiscus were conducted for the first time. The mastic gum and leaves obtained from P. lentiscus may have strong potential in terms of their chemical content and antiviral and cytotoxic activity. As a consequence of these properties, it is a sustainable, renewable natural resource that can be used as an additive and flavoring in the food and pharmaceutical industries. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Chemical profile of the Anatolian Sideritis species with bioactivity studies.

CONTEXT: The genus Sideritis L. (Lamiaceae) is represented by 46 species in Turkey with an 79% endemism ratio, 42 of 46 belonging to the section Empodoclia. OBJECTIVE: In this review article, Sideritis species growing in Turkey have been evaluated for phytochemical constituents and biological activities. METHODS: The data for the isolates, components and extracts of the Anatolian Sideritis species and their bioactivity studies were retrieved from the main databases WoS, Scopus and PubMed from 1975 until 31 December 2022. RESULTS: In this review article, terpenoids, flavonoids, phenolics and other secondary metabolites isolated from Turkish Sideritis species were reported. Anatolian Sideritis species, which primarily consist of monoterpenes and sesquiterpenes, were studied in detail. Sideritis plants are represented by 46 species in Turkey, and 25 of them were investigated for their diterpenoids through isolation or LC-MS studies. Most of the diterpenoids of Turkish Sideritis species have ent-kaurene skeleton, among them linearol, siderol, 7-epicandicandiol and sideridiol were found to be the main compounds. Exceptionally, labdane, pimarane and beyerene diterpenoids were only found in a few species. For phenolics and flavonoids, only 12 species were investigated until now, and they were found to be rich in phenylethanoid glycosides and flavonoid glycosides. In terms of activity, most of the species were tested for antioxidant activity, followed by antimicrobial and anti-ulcer/anti-inflammatory activities. Their cytotoxic, enzyme inhibitory, antinociceptive and antistress activities were less frequently studied. CONCLUSIONS: Sideritis species should be considered promising therapeutic agents in the treatment of upper respiratory tract and ulcer/inflammatory diseases.

Polyphenol Contents, Potential Antioxidant, Anticholinergic and Antidiabetic Properties of Mountain Mint (Cyclotrichium leucotrichum).

In the present work, antioxidant and antidiabetic potentials of mountain mint [Cyclotrichium leu-cotrichum (Stapf ex Rech. Fil.) Leblebici] was the first time appraised. In this sense, methanol (MECL) and water (WECL) extracts were obtained from aerial parts of mountain mint (Cyclotrichium leucotrichum) and studied for their antioxidant ability by several bioanalytical assays. Also, their inhibition profiles were realized toward several metabolic enzymes connected to some diseases, including butyrylcholinesterase (BChE), α-glycosidase, acetylcholinesterase (AChE), and α-amylase enzymes. Additionally, their phenolic contents were determined by putative chromatographic method of LC/MS/MS. Consequently, nineteen phenolic molecules were identified in MECL and fifteen phenolic molecules were found in WECL. Also, antioxidant effects of both extracts were studied using by the methods of 1,1-diphenyl-2-picryl-hydrazyl (DPPH⋅), 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS.+ ) and N,N-dimethyl-p-phenylenediamine (DMPD.+ ) scavenging activities, ferric (Fe3+ ) and cupric (Cu2+ ) ions and Fe3+ -2,4,6-tri(2-pyridyl)-s-triazine (TPTZ) reducing capacities. MECL and WECL were found as powerful DPPH⋅ (IC50 : 23.74 and 28.85 µg/mL), ABTS.+ (IC50 : 12.53 and 14.05 µg/mL) and DMPD.+ scavenging effects (IC50 : 43.52 and 54.80 µg/mL). Also, both extracts demonstrated the effective inhibition on AChE (IC50 : 69.31 and 115.51 µg/mL), BChE (IC50 : 57.75 and 86.62 µg/mL), α-glycosidase (IC50 : 36.47 and 62.94 µg/mL) and α-amylase (IC50 : 1.01 and 3.43 µg/mL). This study will be useful for future studies to determine the antioxidant properties and enzyme inhibition profile of food, medical and industrially important plants.

Identification of natural compounds of Jurinea species by LC-HRMS and GC-FID and their bioactivities.

The Jurinea Cass. is one of the most important genera within Asteraceae and it comprises about 250 species in total. This genus is known for its numerous biological activities such as antioxidant, antimicrobial, antilipid peroxidation, anticholinesterase, antileishmanial activities. The aim of this study was to determine chemical composition and biological activities of ethanol and n-hexane extracts of three different Jurinea species. For this purpose, different parts of J. mollis, J. cadmea and J. pontica were extracted and totally six n-hexane and six ethanol extracts were obtained. Fatty acid content of n-hexane extracts was determined by GC-FID whereas phenolic and flavonoid content of ethanol extracts by LC-HRMS. Palmitic acid (16:0) was detected as the most abundant fatty acid in all n-hexane extracts with the rates ranging from 42.16%-55.08%, except flowers of J. mollis (JMF) and J. cadmea (JCF). LC-HRMS analysis showed the rutin content of all extracts was higher than other flavonoids, except of J. cadmea flowers, whereas apigenin-7-glucoside was found the most abundant in JCF. Cytotoxic effects of the extracts on HeLa and HEK-293 cells were determined by MTT method, and antioxidant activities were evaluated by DPPH and CUPRAC assays. Ethanol extract of J. mollis flowers significantly inhibited cancerous HeLa cells, with the IC50 value of 9.683 µg/mL while it was more less toxic on healthy HEK-293 cells. Ethanol extracts of J. mollis flowers and J. mollis steams-leaves (JMSL) showed the highest antioxidant activity by a DPPH inhibition % of 45.516 ± 2.497 and 56.671 ± 1.496, respectively. JMF and JMSL have also the highest CUPRAC values (0.880 ± 0.067 and 1.085 ± 0.152 mmol TR/g DWE, respectively). Total flavonoid content was determined using aluminum chloride colorimetric assay while total tannin and phenolic content by Folin Chiocalteu's reagent. Results showed that JMSL has the highest total phenolic (108.359 ± 6.241 mg GAE/ G DWE) and flavonoid (32.080 ± 4.385 mg QE/ g DWE) contents whereas JMF has the highest tannin content (121.333 ± 17.889 mg TAE/ g DWE). In the light of these results, various parts of Jurinea species may be regarded as alternative sources for cytotoxic and/or antioxidant flavonoids, phenolics and unsaturated fatty acids that can arouse the interest of pharmaceutical, food and cosmetic industries.

Composition of the essential oil of Satureja metastasiantha: a new species for the flora of Turkey.

The aerial parts of Satureja metastasiantha were hydrodistilled for 3 h using a Clevenger-type apparatus. The essential oils were analyzed by gas chromatography/flame ionization detector and gas chromatography/mass spectrometry, simultaneously, the main compounds of which were characterized as p-cymene (22.3%), thymol (21.0%), carvacrol (18.4%), and γ-terpinene (12.1%). Antioxidant capacity, acetylcholinesterase and butyrylcholinesterase inhibition effects, and antimicrobial and antifungal properties of the species were evaluated. The anticholinesterase activity of the essential oil of S. metastasiantha was observed with 30% inhibition at 200 µg/mL. The essential oil of the species showed activity against Staphylococcus aureus with 128 µg/mL minimum inhibitory concentration value.

Phytochemical content, antioxidant activity, and enzyme inhibition effect of Salvia eriophora Boiss. & Kotschy against acetylcholinesterase, α-amylase, butyrylcholinesterase, and α-glycosidase enzymes.

Many taxa of Salvia genus have been used in herbal beverages, food flavoring, cosmetics, and pharmaceutical industry. In this paper, chemical compounds of Salvia eriophora (S. eriophora) leaves were determined by LC-MS/MS (Liquid Chromatography tandem Mass Spectrometry). Salvigenin (158.64 ± 10.8 mg/kg), fumaric acid (123.09 ± 8.54 mg/kg), and quercetagetin-3.6-dimethylether (37.85 ± 7.09 mg/kg) were detected as major compounds in the ethanol extract, whereas fumaric acid (555.96 ± 38.56 mg/kg), caffeic acid (103.62 ± 20.51 mg/kg), and epicatechin (83.19 ± 8.43 mg/kg) were detected as major compounds in the water extract. Furthermore, enzyme inhibition of S. eriophora against acetylcholinesterase (AChE), α-amylase (AM), butyrylcholinesterase (BChE), and α-glycosidase (AG) enzymes were detected. AChE, BChE, AG, and AM enzymes were very strongly inhibited by S. eriophora water extract (WES) and S. eriophora methanol extract (MES). Additionally, antioxidant potential of S. eriophora was determined by in vitro analytical methods. IC50 values of WES and MES were performed for radicals. PRACTICAL APPLICATIONS: Metabolic enzymes have crucial functions on living systems due to inhibition or activation of them mainly attributed with some health disorders. AChE, BChE, AM, and AG enzymes have important roles on carbohydrate metabolism or cholinergic pathways. The relation between enzyme inhibition effect and phenolic compounds or antioxidant activity need to be confirmed. Thus, many studies tested to clarify this relation for pure samples or plant extracts. To the best of our knowledge, this is the first report about inhibition effects of Salvia eriophora extracts against AChE, BChE, AM, and AG enzymes as well as their phenolic contents and antioxidant activities.

Simultaneous determination of several flavonoids and phenolic compounds in nineteen different Cephalaria species by HPLC-MS/MS.

Cephalaria species in Turkey known as "Pelemir" and they have many different biological activities due to their wide range of chemical content. The main goal of this study is determine the flavonoids and phenolic acids in Cephalaria species using high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). This method was developed for quantitation of reported compounds using reverse phase Fortis C18 (150 × 3 mm × 5 µm) column with a gradient of 0.1% formic acid solution (A) in water and (B) in methanol in ESI mode. The recoveries of developed method were obtained in the range of 90.4-109.4%. Limit of detection (LOD) and limit of quantification (LOQ) values were calculated in the range of 0.3-25.2 ppb and 3.0-102.3 ppb for the reported compounds. Uncertainty evaluations of all compounds were reported. Twenty-five flavonoids, five simple phenolics, two triterpenoids, one iridoid and vitamin C successfully identified and quantified for nineteen Cephalaria species. The main flavonoids in the studied species were determined as cyanidin-3-O-glucoside, luteolin-7-O-glucoside, kaempherol-3-O-glucoside, hyperoside, hesperidin, luteolin and quercetin while salicylic and caffeic acids were detected as major phenolic acids in the analyzed samples. This is the first full characterization study for nineteen Cephalaria species via HPLC-MS/MS.

Phenolic compounds from the aerial parts of Clematis viticella L. and their in vitro anti-inflammatory activities.

Phytochemical investigations on the EtOH extract of Clematis viticella led to the isolation of six flavonoid glycosides, isoorientin (1), isoorientin 3'-O-methyl ether (2), quercetin 7-O-α-L-rhamnopyranoside (3), quercetin 3,7-di-O-α-L-rhamnopyranoside (4), manghaslin (5) and chrysoeriol 7-O-ß-D-glucopyranoside (6), one phenylethanol derivative, hydroxytyrosol (7), along with three phenolic acids, caffeic acid (8), (E)-p-coumaric acid (9) and p-hydroxybenzoic acid (10). The structures of the isolates were elucidated on the basis of NMR and HR-MS data. All compounds were isolated from C. viticella for the first time. Compounds 7 and 8 showed significant anti-inflammatory activity at 100 µM by reducing the release of NO in LPS-stimulated macrophages comparable to positive control indomethacin. Compounds 3 and 7 exhibited anti-inflammatory activity through lowering the levels of TNF-α while 1, 3 and 5 decreased the levels of neopterin better than the positive controls.

Different propolis samples, phenolic content, and breast cancer cell lines: Variable cytotoxicity ranging from ineffective to potent.

Researchers have started focusing on investigating the anticarcinogenic effects of natural products with the slightest side effects possible, because current breast cancer treatment approaches are unable to achieve absolute success especially on aggressive subtypes. Propolis is among these products with its antimicrobial, antifungal, anti-inflammatory, and anticancer effects. Therefore, seven different samples were collected from different regions (Argentina, China, and Istanbul-Turkey) and applied on nonaggressive breast cancer cell line (BCCL) MCF-7 and aggressive cell lines SK-BR-3, and MDA-MB-231. Initially, the phenolic/flavonoid constituents of the propolis ethanol extracts were investigated by liquid chromatography-mass spectrometry-mass spectrometry (LS-MS/MS) and high-performance liquid chromatography (HPLC) analyses. Then, the anticarcinogenic effects of the propolis samples on MCF-7, SK-BR-3, MDA-MB-231 were evaluated by WST1 analysis and only selected ones on MCF-10A and hPdLF. According to the LS-MS/MS and HPLC analysis, Turkey originated propolis (Turkey3) were found to be richer than the other propolis samples in terms of phenolic/flavonoid compounds. Turkey propolis significantly inhibited cell proliferation in both nonaggressive and aggressive BCCL (P < 0.01). Therefore, Turkey3 propolis was selected for further evaluation using Annexin V-PI apoptosis detection assays. In addition, selected compounds among the propolis contents such as galangin, caffeic acid, apigenin, quercetin, and ferulic acid were applied to the MCF-7 cell line to detect cytotoxic and apoptotic effects. Galangin, caffeic acid, apigenin, and quercetin remarkably induced cell proliferation inhibition at all time intervals, whereas ferulic acid was found non efficient on the MCF-7 cell line. Annexin V-PI assay clarified that all cell proliferation inhibitions were markedly apoptotic. Our findings indicated that the inhibition effect of propolis on breast cancer cell proliferation was in a propolis type-, dose- and time-dependent fashion. Turkey3 propolis showed statistically significant cytotoxic effects on both the nonaggressive and aggressive BCCL. These findings were consistent with the effects of its rich phenolic and flavonoid contents, in terms of variety. © 2018 IUBMB Life, 71(5):619-631, 2019.

Development and validation of a generic method for quantification of collagen in food supplement tablets using liquid chromatography coupled with time-of-flight mass spectrometry.

A generic method for the quantification of type II collagen in protein-based dietary supplements is described. This quantitative analysis was conducted using liquid chromatography-electrospray ionization-time-of-flight mass spectrometry (LC-ESI-TOF MS). Compared to classical methods with the use of isotope-labeled standards, our method includes, for the first time, the quantification of hydroxyproline using histidine as an internal standard. Separation of the analytes was performed on a Phenomenex Synergi 4 µm Fusion-RP 80 Ǻ column (150 × 2.0 mm, 4.0 µm) with a mobile phase made of 10 mM ammonium formate in water (A) and 10 mM ammonium formate in methanol (B). The assay was fully validated according to FDA guidelines, and the method exhibited sufficient specificity, accuracy, and precision. Intra- and inter-batch accuracy, determined as a deviation between nominal and measured values, ranged from -4.8 to 9.1% and from 0.9 to 6.4 %, respectively. All analytes (hydroxyproline and histidine) at three concentration levels showed extraction recoveries from 89 to 98 %. The method was successfully applied to protein-based dietary supplements of the pharmaceutical industry.

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.

Isolation of active constituents from cherry laurel (Laurocerasus officinalis Roem.) leaves through bioassay-guided procedures.

ETHNOPHARMACOLOGICAL RELEVANCE: The fresh leaves of Laurocerasus officinalis Roem. (Rosaceae) are externally used against pain and feverish symptoms in Turkish folk medicine. AIM OF THE STUDY: Effects of the extracts, fractions and isolated compounds from the leaves of L. officinalis were investigated using in vivo models of inflammation and pain in mice. METHODS: The crude ethanolic extract from the leaves of plant was sequentially fractionated into five subextracts; explicitly, n-hexane, chloroform, ethyl acetate (EtOAc), n-butanol, and remaining water extracts. Further studies were carried out on the most active EtOAc subextract was further subjected to fractionation through column chromatography. For the anti-inflammatory activity, carrageenan-induced hind paw edema and acetic acid-induced increase in capillary permeability models, and for the antinociceptive activity p-benzoquinone-induced writhing test in mice were employed. RESULTS: Ethanolic extract of the leaves was shown to possess significant inhibitory activity in the assay methods without inducing any gastric damage. Through bioassay-guided fractionation and isolation procedures three phenolic compounds, 2-O-ß-D-glucopyranosyl-2-hydroxyphenyl-acetic acid (1), kaempferol-3-O-ß-D-xylopyranosyl-(1→2)-O-ß-D-glucopyranoside (2) and (+)-catechin (3) were isolated from the active fraction and their structures were elucidated by spectral techniques (1D and 2D NMR, ESIMS). CONCLUSION: The experimental data verified that Laurocerasus officinalis leaves displayed remarkable anti-inflammatory and antinociceptive activity.

Pomological features, nutritional quality, polyphenol content analysis, and antioxidant properties of domesticated and 3 wild ecotype forms of raspberries (Rubus idaeus L.).

The raspberry (Rubus idaeus L.) is an economically important berry crop that contains many phenolic compounds with potential health benefits. In this study, important pomological features, including nutrient content and antioxidant properties, of a domesticated and 3 wild (Yayla, Yavuzlar, and Yedigöl) raspberry fruits were evaluated. Also, the amount of total phenolics and flavonoids in lyophilized aqueous extracts of domesticated and wild ecotypes of raspberry fruits were calculated as gallic acid equivalents (GAEs) and quercetin equivalents (QE). The highest phenolic compounds were found in wild Yayla ecotype (26.66 ± 3.26 GAE/mg extract). Whilst, the highest flavonoids were determined in wild Yedigöl ecotype (6.09 ± 1.21 QA/mg extract). The antioxidant activity of lyophilized aqueous extracts of domesticated and wild ecotypes of raspberry fruits were investigated as trolox equivalents using different in vitro assays including DPPH(•), ABTS(•+), DMPD(•+), and O(•-)(2) radical scavenging activities, H(2)O(2) scavenging activity, ferric (Fe(3+)) and cupric ions (Cu(2+)) reducing abilities, ferrous ions (Fe(2+)) chelating activity. In addition, quantitative amounts of caffeic acid, ferulic acid, syringic acid, ellagic acid, quercetin, α-tocopherol, pyrogallol, p-hydroxybenzoic acid, vanillin, p-coumaric acid, gallic acid, and ascorbic acid in lyophilized aqueous extracts of domesticated and wild ecotypes of raspberry fruits were detected by high-performance liquid chromatography and tandem mass spectrometry (LC-MS-MS). The results clearly show that p-coumaric acid is the main phenolic acid responsible for the antioxidant and radical scavenging activity of lyophilized aqueous extracts of domesticated and wild ecotypes of raspberry fruits.

Chemical composition of natural colophony from Pinus brutia and comparison with synthetic colophony.

The compositions of colophony resins obtained from Pinus brutia Ten trees by three different methods (acid paste, carved hole and scraping) from Ayvacik, Gökova and Kemalpasa in Turkey were analyzed by capillary GC-MS. The main components were the monoterpenes alpha-pinene, beta-pinene, and delta3-carene, and the diterpenic resin acids palustric, abietic, kaur-9(11)-16-en-18-oic and neoabietic acid. The synthetic colophony resins exhibited similar contents to those of the natural resins obtained from the Gökova and Kemalpasa regions of Turkey. However, colophony resins from Ayvacik exhibited only half the diterpenic acid content as those of the Gökova and Kemalpasa resins. Out of the three techniques, the carved hole method caused rather different percentages in the constituents of the essential oils.

DNA damaging activities of methanol extract of Ajuga postii and iridoid glucoside reptoside.

An iridoid glucoside reptoside (1) has been isolated as a DNA damaging active agent by bioassay-guided fractionation of the methanol extract of Ajuga postii. Furthermore, from the acetone extract of A. postii two known triterpenic compounds ursolic acid, alpha-amyrin and two steroidal compounds (24S)-24-ethylcholesta-5,25-dien-3beta-ol and beta-sitosterol were isolated. Their structures were elucidated based on 1D and 2D NMR techniques and mass data.

Cytotoxic and DNA damaging activity of some aporphine alkaloids from Stephania dinklagei.

Six aporphine alkaloids, liriodenine, corydine, isocorydine, atherospermidine, stephalagine and dehydrostephalagine, were isolated by bioassay-guided fractionation of the EtOH extract of the stems of Stephania dinklagei. Liriodenine showed strong cytotoxic activity while corydine and atherospermidine showed DNA damaging activity. Dehydrostephalagine and atherospermidine were isolated from this plant for the first time.

Diterpenes from Sideritis trojana.

Six known ent-kaurene, a new ent-kaurane and a new pimarane diterpenes were isolated from Sideritis trojana. The structures of new compounds were determined as ent-7alpha-15beta,16beta-epoxykaurane (1), and ent-2alpha-hydroxy-8(14),15-pimaradiene (2) along with the known compounds siderol (3), sideridiol (4), 7-epicandicandiol (5), isocandol B (6), candol A acetate (7) ent-7alpha-acetoxykaur-15-ene (8) by IR, 1D and 2D NMR techniques and HRMS.

Diterpenoids from Salvia ceratophylla.

Salvia ceratophylla L. has yielded four known and two new diterpenoids together with two triterpenic acids, a steroid and a flavone. The structures of the compounds were established by spectroscopic analyses. One of the known compounds candidissiol exhibited a high antibacterial activity against Staphylococcus epidermidis and Proteus mirabilis.