Q-TOF Analysis Research Articles

Citrus wastewater as a source of value-added products: Quali-quantitative analysis and in vitro screening on breast cancer cell lines.

Citrus wastewater from industries is a source of bioactive compounds whose recovery could be a useful approach to convert processing waste into potential resources to be exploited in food, pharmaceutical, and chemical companies. Citrus wastewater, obtained from the industrial processing of Citrus sinensis, was freeze-dried and qualitative/quantitative evaluated using HPLC/MS Q-TOF analysis. Antiproliferative activity was investigated on MDA-MB-231 (triple-negative breast cancer cell line), MCF-7 (breast cancer cell line), and its multidrug-resistant variant MCF-7R. Fraction 8 emerged for its cytotoxicity toward MCF-7R cells. Its main component, the polymethoxylated flavone nobiletin (80%), is likely involved in increasing the number of G1-phase MCF-7R cells without inducing cell death. Notably, fraction 8 sensitizes MCF7-R cells to the antiproliferative effects of doxorubicin, thus contributing to overcoming MCF7-R multidrug resistance. Our studies highlighted the possibility of applying a sustainable strategy for citrus wastewater recycling to recover functional compounds as useful adjuvants for the prevention and treatment of malignancies.

EXPLORING THE HAEMOSTATIC POTENTIAL OF PANCHAVALKALA KASHAYA AND PANCHAVALKALA AQUEOUS EXTRACT: AN IN VITRO STUDY

Acharya Susrutha under the context of Rakthasthambhanopaya (methods of haemostasis) has explained about the different methods to attain Rakthasthambhana or haemostasis. which include Skandana (clotting of blood), Santhana (uniting), Pachana (ripening or digestive measures) and Dahana (thermal cauterisation). Acharya Susrutha while explaining about agantuja vrana chikitsa (treatment of wound) has explained that ‘Pittavat sheeta kriya’ should be done prior to santhana to mitigate the ‘Kshatoshmana’. So, from this it is evident the primary method of management should be done with sheeta (cold) followed by Kashaya rasa dravya (astringent taste drugs) for Santhana. Panchavalkala is a group drug with sheeta veerya (cold potency) and Kashaya rasa (astringent taste) mentioned in Dalhana commentary for Skandana. This study investigates the haemostatic effect of Panchavalkala Kashaya and Panchavalkala aqueous extract by in vitro method. Preliminary phytochemical screening of both trial drugs was done. A total of 10 samples of healthy human platelet poor plasma were mixed in trial drugs, Prothrombin and Activated Partial Thromboplastin time were determined. Data was analysed using Kruskal Wallis test. Phytochemical analysis confirmed that the Panchavalkala kashaya contain alkaloids, tannins, glycosides and saponins. Panchavalkala aqueous extract sample consist of flavonoids, alkaloids, glycosides, tannins and saponins. LCMS Q-TOF analysis shows the presence of compounds Gallic acid, Epicatechin, Leucocynidin and Procyanidin B1. And the HPLC study confirmed that Gallic acid is an active phytoconstituent present in the Panchavalkala aqueous extract. In the present study the trial group Panchavalkala aqueous extract showed more desirable haemostatic effect than Panchavalkala Kashaya on comparison.

Metabolites from a Pleurotus tuber-regium collected in Madagascar

Pleurotus tuber-regium was isolated from a dead trunk of Raphia farinifera (Arecaceae) in a lowland moist forest in Antsohihy, Madagascar, and the species was confirmed by molecular analysis and morphological observations. The main bioactive metabolites of the mycelium extracts were identified by mass spectrometry techniques. Five structural diverse metabolites, tryptophol, pyroglutamic acid, prolyldiketopiperazine B, sporol and RKS-1778, were characterised by LC-MS qTOF analysis of the hydro-alcoholic extract. GC-MS analysis of both chloroform and ethyl acetate extracts revealed the presence of several saturated and ­unsaturated fatty acids and their esters derivatives.

N, S co-doped porous carbon to accelerate Fe3+/Fe2+ redox cycle for peroxymonosulfate activation

Fenton-like process has important implications for water treatment due to its effective degradation of refractory contaminants. However, sluggish conversion of Fe3+ back to Fe2+ strongly restricts its practical application. Herein, a novel N, S co-doped porous carbon (S-ZCN) derived from ZIF-8 was prepared as a metal-free heterogeneous catalyst to accelerate the redox cycle of Fe3+/Fe2+, leading to more efficient peroxymonosulfate (PMS) activation. With only 20 mg/L S-ZCN added, sulfamethoxazole (SMX) could be removed completely by the S-ZCN/Fe3+/PMS system within 40 min. Reactive oxygen species (ROS) were identified by quenching experiments, electron paramagnetic resonance (EPR), and probe experiments. It was revealed that hydroxyl radical (HO•) was the primary ROS, which contributed 79.2 % to SMX degradation. Verified by Fe concentration detection, chelating agent experiments and the results of XPS and FT-IR spectra, the synergistic effect of N, S co-doping and formation of Fe-Nx sites promoted the electron transfer from sp2-hybridized C to surface-associated Fe3+ to reconstruct Fe2+/PMS system. Based on the detection results of intermediates by Q-TOF analysis, potential degradation pathways for SMX in the S-ZCN/Fe3+/PMS system were proposed. Wide range of pH applicability and strong degradation to various pollutants exhibited the advantages in practical applications. This work provides a novel approach involving metal-free catalysts to break the bottleneck in the field of traditional Fenton-like reaction.

Comprehensive Phytochemical Analysis and Bioactivity Evaluation of Padina boergesenii: Unveiling Its Prospects as a Promising Cosmetic Component.

Marine macroalgae, such as Padina boergesenii, are gaining recognition in the cosmetics industry as valuable sources of natural bioactive compounds. This study aimed to investigate the biochemical profile of P. boergesenii and evaluate its potential as a cosmetic ingredient. Fourier-transform infrared (FTIR), gas chromatography-mass spectrometry (GCMS), and high-resolution liquid chromatography-mass spectrometry quadrupole time-of-flight (HRLCMS QTOF) analyses were employed to assess the functional groups, phycocompounds, and beneficial compounds present in P. boergesenii. Pigment estimation, total phenol and protein content determination, DPPH antioxidant analysis, and tyrosinase inhibition assay were conducted to evaluate the extracts' ability to counteract oxidative stress and address hyperpigmentation concerns. Elemental composition and amino acid quantification were determined using inductively coupled plasma atomic emission spectrometry (ICP-AES) and HRLCMS, respectively. FTIR spectroscopy confirmed diverse functional groups, including halo compounds, alcohols, esters, amines, and acids. GCMS analysis identified moisturizing, conditioning, and anti-aging compounds such as long-chain fatty alcohols, fatty esters, fatty acids, and hydrocarbon derivatives. HRLCMS QTOF analysis revealed phenolic compounds, fatty acid derivatives, peptides, terpenoids, and amino acids with antioxidant, anti-inflammatory, and skin-nourishing properties. Elemental analysis indicated varying concentrations of elements, with silicon (Si) being the most abundant and copper (Cu) being the least abundant. The total phenol content was 86.50 µg/mL, suggesting the presence of antioxidants. The total protein content was 113.72 µg/mL, indicating nourishing and rejuvenating effects. The ethanolic extract exhibited an IC50 value of 36.75 μg/mL in the DPPH assay, indicating significant antioxidant activity. The methanolic extract showed an IC50 value of 42.784 μg/mL. Furthermore, P. boergesenii extracts demonstrated 62.14% inhibition of tyrosinase activity. This comprehensive analysis underscores the potential of P. boergesenii as an effective cosmetic ingredient for enhancing skin health. Given the increasing use of seaweed-based bioactive components in cosmetics, further exploration of P. boergesenii's potential in the cosmetics industry is warranted to leverage its valuable properties.

Prediction of Fragmentation Pathway of Azilsartan Protonated Ion Using High Resolution Mass Spectrometry (HR-MS/MS) with Q-TOF Analyzer- A Perspective Study

Introduction: UPLC coupled with a high-resolution mass analyzer i.e, Q-TOF analyzer with electrospray ionization (ESI) source using collision-induced dissociation (CID) method was applied to study the characteristic product ions of azilsartan protonated ion. Method: The experimental results of high-resolution mass spectra explained the elemental compositions of product ions accurately and reasonable fragmentation pathways were proposed for azilsartan protonated ions. Calculated mass error in parts per million (ppm) for molecular ions and product ions and results of mass error found from this study from 0.2 ppm to 3.5 ppm. Result: The characteristic fragmentation pathways were helpful to analyze and interpret the stability and possible degradation pathway of the parent ion. Conclusion: The present study explains the significant role of high-resolution mass spectrometry in the structural analysis of the protonated ion of azilsartan.

Oroxylum indicum Vent Root Bark Extract Inhibits the Proliferation of Cancer Cells and Induce Apoptotic Cell Death

Oroxylum indicum Vent is a medium-sized deciduous tree that belongs to the family Bignoniaceae. The roots of this tree are used as one of the ten ingredients to prepare the Dasamula formulation in the Ayurvedic system of medicine in India. Although traditional uses are attributed to the medicinal plant, there are limited scientific data on its potential. The present study thus analyzed the cytotoxic and apoptotic potential of the plant against different cancer cells. MTT assay was used to determine cytotoxicity using HeLa, HCT 15, and MDA-MB-231 cells, with the IC50 values, revealed at concentrations of 92.43, 133.0, and 112.84 µg/mL respectively. However, the extract was less toxic to non-cancer cells. HeLa cells further treated with OIM were subjected to flow cytometric analysis for studying the cell cycle stages. When untreated cells at G1 phase were found at a relative percentage of 71.9%, it increased to 79.3 and 86% with OIM treatment at concentrations of 50 and 100 µg/mL; cells in the S phase decreased from 10.3 to 8.2 and 7.5%, concluding the arrest of the cell cycle at G1 phase. With further study of apoptotic morphology with dual acridine orange–ethidium bromide staining and Annexin–Hoechst staining, cells at early and late apoptotic stages were observed with OIM treatment at 100 µg/mL concentration. Although such effects were noticed with OIM treatment, it could not be concluded that the extract had remarkable anti-proliferative effects, since the small changes noticed in cell cycle arrest and apoptotic induction were attained at a high concentration of OIM 100 µg/mL. The biological activities of plants and their extracts are attributed to the presence of multifarious compounds present in them. LC-MS Q-TOF analysis confirmed the presence of biochanin A and baicalein in OIM. HPLC-based quantification of baicalein and chrysin was shown to be 3.36 and 1.11 mg/gram dry weight. To conclude, the above results suggest that the root bark of O. indicum has a broad spectrum of biological activities, including anticancer and apoptotic properties.

Effect of Apple Juice Enrichment with Selected Plant Materials: Focus on Bioactive Compounds and Antioxidant Activity.

Using a multi-analytical approach, this paper aimed to investigate the effect of apple juice enrichment with Arbutus unedo and Diospyros kaki fruits, Myrtus communis berry extract, Acca sellowiana, or Crocus sativus flower by-products on both bioactive compounds content and antioxidant activity. Physico-chemical parameters, vitamin C, sugars, organic acids, total polyphenol content, antioxidant activity, and sensory attributes were evaluated. An LC-PDA/MS QTof analysis allowed for the identification of 80 different phenolic compounds. The highest polyphenol content (179.84 and 194.06 mg of GAE/100 g fw) and antioxidant activity (CUPRAC, 6.01 and 7.04 mmol of Fe2+/100 g fw) were observed in products with added A. sellowiana and D. kaki, respectively. Furthermore, the study showed a positive correlation between polymeric procyanidins and antioxidant activity (0.7646-0.8539). The addition of A. unedo fruits had a positively significant influence on the increment of vitamin C (23.68 ± 0.23 mg/100 g fw). The obtained products were attractive to consumers, especially those with 0.1% C. sativus flower juice, M. communis berry extract, and persimmon D. kaki fruits. The synergy among the different analytical techniques allowed us to obtain a complete set of information, demonstrating that the new apple smoothies were enriched in both different beneficial molecules for human health and in antioxidant activity.

Anticancer, Antioxidant, Antimicrobial and Enzyme Inhibitory Activities of Inula aucheriana

Inula aucheriana showing widespread in Turkey, it is a plant belonging to the Asteraceae family. It is aimed to reveal the pharmacological potential of this plant with this study, which investigates the different biological properties of this plant, which is known to be used in the treatment of various diseases. Methods: 80% ethanol extract of I.aucheriana was prepared and investigated the chemical composition (Q-TOF analysis), antioxidant (spectrophotometric analysis), enzyme inhibitory activities (Ellman's method), antimicrobial (MIC concentration value), anticancer (IC50 analysis) activities with qualitative and quantitative methods. Results: 80% ethanol extract of I.aucheriana was showed that is a powerful antioxidant, anticancer and enzyme inhibitor. In the chemical composition analysis, the major compound of the exctract was determined as luteolin (32.55%). I. aucheriana extract has been shown to have AChE and BChE inhibition. In addition, when the Inula extract was compared with the reference drug, it was determined that the extract had an inhibitory effect in terms of ɑ-glucosidase. It also showed quite high tyrosinase enzyme inhibition. When its antiproliferative activity was examined, it was found to be quite active in the MDA-MB-231 cell line. Conclusion: This study is the first investigation of the enzyme inhibitory activity and antioxidant activity of I. aucheriana. According to the data obtained from the study, more detailed studies can be conducted with the active ingredients of I. aucheriana, and promising results can be obtained for the treatment of different diseases.

Biological and phytochemical characterization of Phragmanthera regularis (Loranthaceae) from Ethiopia

Phragmanthera regularis is a hemi-parasitic shrub. It is known for treating various health ailments. The aim of this study was to evaluate the antimicrobial activity, toxicity, and chemical characterization of the leaf extracts of P regularis collected from the Schinus molle host plant in Ethiopia. The antimicrobial properties of crude extracts obtained with chloroform, ethyl acetate, methanol, and water solvents were assayed against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. The methanol extract significantly inhibited the growth of S. aureus, E. coli and P. aeruginosa were resistant to any of these solvent extracts. The methanol extract was tested at 175, 550, and 2000 mg/kg body weight doses in white mice and did not reveal any toxicity. The LC-MS qTOF analysis detected flavonoids, phenolic acids, and alkaloids in the crude methanol extract. Further study is needed to investigate the effectiveness of these compounds against S. aureus.

GC-MS and LC-MS/MS Analysis of Bouea macrophylla Fruit Juice

Bouea macrophylla Griff is a member of the Anacardiaceae family, which has edible fruits. Due to the high nutritional value, appealing taste, and attractive color, the fruit juice of B. macrophylla has the potential to be developed as a functional beverage. The present study was conducted to determine the phytochemical profile of B. macrophylla fruit juice using GC-MS and LC-MS/MS. GC-MS analysis was performed with the Agilent Technologies 7890 Gas Chromatograph with automated sampling and tandem Agilent 5975 Mass Selective Detector and peak analysis with the Chemistation data system. LC-MS/MS QTOF analysis was performed with UPLC Waters I Class and tandem MS/MS Xevo G2S QTOF with UNIFI software. GC-MS analysis identified five compounds with a quality match of 90% and above, namely 3-pentadecylphenol (31.68%), 5,5’-(oxybis(methylene)]-bis-2-furancarboxy-aldehyde (5.67%), 5-(hydroxymethyl)-2-furancarboxy-aldehyde (23.51%), 4H-pyran-4-one, 2,3-dihydro-3,5-dihydro-6-methyl (5.00%), and n-hexadecanoic acid (4.71%). Five major compounds were identified for the first time in B. macrophylla fruit juice by LC-MS/MS analysis, i.e., (E)-hexadecyl-ferulate, digupigan A, eburicoic acid, hydroginkgolinic acid, and terminalic acid. The health benefit of some of these detected phytochemicals supports the use of juice as the main ingredient for functional food.

Hydrogen-deuterium (H/D) exchange reaction of acebutolol hydrochloride in D₂O and CD₃OD solution.

H/D exchange reactions can be observed by NMR spectroscopy of acebutolol (ACE). The results obtained showed deuterium incorporation at α-posi t ion of the carbonyl group of acebutolol, when using deuterium oxide or deuterated methanol as deuterium source and solvent. The spontaneous deuteration is proceeded by the following pathway CH₃→CH₂D→CHD→CD₃, through a keto-enol tautomerization reaction. Furthermore, LC-MS / QTOF analyses have confirmed the proposed H/D exchange. In order to reduce the time of total deuteration observed at the acetyl group alkaline catalysts were employed.

Estrogenic Effect of Gastrodia elata Blume Extract

Background: A decline in the function of the ovaries and a decrease in the production of sex hormones with aging causes hormonal deficiencies, leading to female menopause. Natural products may be developed into estrogen-like substances to improve menopausal conditions in women.BRMethods and Results: We investigated whether Gastrodia elata Blume (GEB) extract could produce a similar effect to estrogen (E2) on MCF-7 cells, from a female breast cancer cell line. Trefoil factor 1 (TFF-1) and progesterone receptor (PR) were upregulated in the GEB extract-treated cells, like the E2 treatment. Moreover, the levels were upregulated in the phosphorylated mitogen-activated protein (MAP) kinase kinases 1/2, extracellular signal-regulated kinase 1/2, estrogen receptor (ER) alpha, and ER beta. An ER antagonist (ICI 182.780) with the GEB extract induceds a downstream response via the ER pathway. A qTOF analysis using a standard chemical compound revealed bis(4-hydroxybenzyl)ether was an effective compound.BRConclusions: GEB extract had an estrogen-like effects in vitro and bis(4-hydroxybenzyl)ether was an effective compound.

Disparities in chemical exposures among pregnant women and neonates by socioeconomic and demographic characteristics: A nontargeted approach

BackgroundExposure to environmental chemicals during pregnancy adversely affects maternal and infant health, and identifying socio-demographic differences in exposures can inform contributions to health inequities. MethodsWe recruited 294 demographically diverse pregnant participants in San Francisco from the Mission Bay/Moffit Long (MB/ML) hospitals, which serve a primarily higher income population, and Zuckerberg San Francisco General Hospital (ZSFGH), which serves a lower income population. We collected maternal and cord sera, which we screened for 2420 unique formulas and their isomers using high-resolution mass spectrometry using LC-QTOF/MS. We assessed differences in chemical abundances across socioeconomic and demographic groups using linear regression adjusting for false discovery rate. ResultsOur participants were racially diverse (31% Latinx, 16% Asian/Pacific Islander, 5% Black, 5% other or multi-race, and 43% white). A substantial portion experienced financial strain (28%) and food insecurity (20%) during pregnancy. We observed significant abundance differences in maternal (9 chemicals) and cord sera (39 chemicals) between participants who delivered at the MB/ML hospitals versus ZSFGH. Of the 39 chemical features differentially detected in cord blood, 18 were present in pesticides, one per- or poly-fluoroalkyl substance (PFAS), 21 in plasticizers, 24 in cosmetics, and 17 in pharmaceuticals; 4 chemical features had unknown sources. A chemical feature annotated as 2,4-dichlorophenol had higher abundances among Latinx compared to white participants, those delivering at ZSFGH compared to MB/ML, those with food insecurity, and those with financial strain. Post-hoc QTOF analyses indicated the chemical feature was either 2,4-dichlorophenol or 2,5-dichlorophenol, both of which have potential endocrine-disrupting effects. ConclusionsChemical exposures differed between delivery hospitals, likely due to underlying social conditions faced by populations served. Differential exposures to 2,4-dichlorophenol or 2,5-dichlorophenol may contribute to disparities in adverse outcomes.

Enhanced oxidation of fluoroquinolones by visible light-induced peroxydisulfate: The significance of excited triplet state species

Recently, catalyst-free activation of peroxydisulfate (PDS) via visible light has been extensively investigated. However, the intrinsic relationship between the activation of PDS and the characteristics of pollutants has been largely ignored. This study reports that PDS activation by visible light without any artificial catalyst for the removal of fluoroquinolones (FQs), which reduced the adverse on ecological environment and human health concerns. Importantly, the mechanism of PDS activation by the excited triplet state of FQs (3FQs*) was proposed. Experimental results demonstrated PDS could generate more ∙OH with the existence of 3FQs*. Meanwhile, the electron transfer pathways from FQs or 3FQs* to PDS were studied. Both radical and non-radical oxidation reactions lead to the degradation of FQs in the Vis/PDS system. The degradation pathways and reactive sites were clarified by QTOF analysis and DFT calculations. Additionally, the degradation experiments of various contaminants demonstrated that the system showed excellent selective oxidation for FQs.

Lipidomic profiling of Pleurotus ostreatus by LC/MS Q-TOF analysis

In this paper, for the first time a lipidomic analysis on Pleurotus ostreatus species was performed by liquid chromatography quadrupole time-of-flight mass spectrometry (LC/MS Q-TOF). Twenty-seven lipid classes, including polar and non-polar lipid classes, were detected. Free fatty acids (FA) were the predominant fraction (>57%), followed by fatty acid ester of hydroxyl fatty acid and ceramide. C18 chain length and two double bonds were the main structural characteristics for FA. Phosphatydilcholine, phosphatydiletanolamine, and glycerophosphates showed high percentages of polyunsaturated fatty acids. Unconventional fatty acids, such as odd and oxygenated chains, were detected. The highest odd/even ratio was found in hexosylceramides and sphingomyelin, while oxygenated chains were mainly represented in ceramides. As a preliminary approach, the results of lipid molecular species, subjected to principal component analysis and discriminant analysis, were able to differentiate P. ostreatus samples on the base of grown substrate. The results of the comprehensive analysis of P. ostreatus lipids are useful to evaluate the lipid nutritional value and could facilitate exploitation of P. ostreatus consumption.

Polyphenols in the Waste Water Produced during the Hydrodistillation of ‘Narcea Roses’ Cultivated in the Cibea River Valley (Northern Spain)

The ‘Narcea rose’ is a recently described yet ancient rose cultivar of interest to the perfume industry. Given its excellent adaptation to the conditions of the place where it was rediscovered, the possibilities of its horticultural/industrial production have been under examination for some time. The hydrodistillation process produces a red-to-brownish mixture of water and rose petals that could contain compounds that could be used in other industrial procedures. Their recovery and further utilization would reduce disposal costs and improve the sustainability of relevant industries. This work reports the quantification, by high-performance liquid chromatography (HPLC–MS) and quadrupole time of flight Q-TOF analyses, of the polyphenol content in the waste water. This waste was found to contain high concentrations of quercetin, gallic acid and ellagic acid, as well as smaller concentrations of kaempferol and its derivatives, all of which can influence plant, human and animal health.

Recovery of antioxidant from Decapterus Macarellus waste using wet rendering method

In Malaysia, Decapterus Maruadsi is widely used as the main ingredient in food products such as ‘keropok lekor’ and fish cracker. The wastes generated from the fish snack industry are underutilized and most are discarded into the sea, generating considerable environmental pollution. The recovery of antioxidants in the fish wastes could lead to the development of a very exciting and promising alternative source of commercial antioxidant compounds. The objective of the present work is to investigate the effect of temperature, type of solvent, extraction time, and ratio of sample to solvent loading in the extraction of antioxidants from Decapterus Maruadsi waste using the wet rendering method. The collected fish waste of Decapterus Maruadsi was pre-treated and then subjected to a wet rendering treatment at 0–60 °C for 30–150 min with ratio of sample to solvent loading of between 1:10 and 9:10 (w/v). The presence of antioxidant in the extract was analyzed using 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and Total Phenolic Compound (TPC) assays. Based on the results of this study, wet rendering treatment performed for 90 min at 30 °C with a ratio of sample to solvent loading of 3:10 produced an extract with the highest concentration of active antioxidant compounds. Besides, only N-Acetyl-glutamic acid was identified with LCMS QTOF analysis. These results suggest that the fish waste of Decapterus Maruadsi can be exploited to become a source of antioxidants for food and pharmaceutical applications.

Propolis Extract Regulates microRNA Expression in Glioblastoma and Brain Cancer Stem Cells.

Grade IV gliomas are classified as glioblastoma (GBM), which is the most malignant brain cancer type. Various genetic and epigenetic mechanisms play a role in the initiation and progression of GBM. MicroRNAs (miRNAs) are small, non-coding RNA molecules that belong to the main epigenetic regulatory RNA class that plays different roles in either physiological or pathological conditions, including GBM pathogenesis regulating expression levels of the target genes. Brain Cancer Stem Cells (BCSCs) are responsible for poor prognosis, including therapy resistance and relapse. Epigenetic regulation mediated by miRNAs is also a critical component of BCSC selfrenewal and differentiation properties. Propolis is a resinous substance collected by honey bees from various plant sources. The flavonoid content of propolis varies depending on the collection region and the extraction method. Although there are studies that include the effects of different originated-propolis on the miRNA expression levels of the glioblastoma cells, the impact on the BCSCs has not been studied yet. This study aims to evaluate the effects of propolis obtained from Aydın, a city in western Turkey, on miRNA expression levels of BCSCs and GBM cells. Aydin propolis was dissolved in 60% ethanol, and after evaporation, distilled water was added to prepare the propolis stock solution. The flavonoids content of the Aydin propolis was determined by MS Q-TOF analysis. Commercially obtained U87MG and BCSCs were used as in-vitro brain cancer models. Cytotoxic and apoptotic effects of Aydın propolis were determined via WST-1 assay and Annexin V test, respectively. The miRNA expression profile was investigated using the real-time qRT-PCR method. The fold changes were calculated by the2-ΔΔCt method. The miRNA-mRNA-pathway interactions, including significantly altered miRNAs, were determined using different bioinformatics tools and databases. Quercetin 3-methyl ether was the main component of the Aydin propolis. Aydin propolis did not show significant cytotoxic and apoptotic effects on both GBM and BCSCs up to 2mg/ml concentration. Aydin propolis treatment decreased the expression of nine miRNAs in the U87MG and five miRNAs in the BCSCs. Moreover, ten miRNAs have upregulated from 2.22 to 10.56 folds in propolis treated GBM cells compared to the control group significantly (p<0.05). In the study, the potential roles of two new miRNAs, whose regulations in glioma were not previously defined, were identified. One of them was miR-30d-5p, a novel potential oncomiR in GBM, which was 2.46 folds downregulated in Aydin propolis treated GBM cells. The other one is miR-335-5p, which is a potential tumor suppressor miR in GBM, that was 5.66 folds upregulated in Aydin propolis treated GBM cells. FOXO pathway, its upstream and downstream regulators, and critically neuronal developmental regulators, NOTCH and WNT pathways, were determined as the most deregulated pathways in Aydin propolis treated cells. The determination of the anti-cancer effect of Aydın propolis on the miRNA expression of GBM, especially on cancer stem cells, may contribute to the elucidation of brain cancer genetics by supporting further analyses.

Qualitative LC-Q-TOF Analysis of Umbilical Cord Tissue via Data-Dependent Acquisition as an Indicator of In Utero Exposure to Toxic Adulterating Substances.

Toxic adulterants are drug or chemical agents used to add bulk volume to traditional drugs of abuse such as cocaine and heroin. These cutting agents include levamisole, metamizole, noxiptillin, phenacetin and xylazine as well as common legal drugs such as acetaminophen, caffeine, diphenhydramine, lidocaine, quinine, quetiapine and tramadol. Because they possess pharmacological activity they result in exposure of the user, but also in the case of pregnant women, the developing fetus, to potential drug toxicity. We describe the development, validation and implementation of a rapid (48 second sample-to-sample) test based on a qualitative data-dependent liquid chromatography-quadrupole time of flight mass spectrometry method for the analysis of toxic adulterating substances in umbilical cord tissue (UCT) samples. The method provides a means of studying potential in utero exposure to these agents. Library spectra comparison at three different collision energies was used in conjunction with retention time and accurate mass to identify these substances in UCT. Analytically based reporting limits were established to determine positivity rates of adulterants in UCT utilizing a standard addition approach. The method was applied to authentic cocaine and opioid positive UCT to screen for toxic adulterants. There were a total of 82 potential adulterant positives found in a 30-sample cohort of authentic UCT samples, with an average of 2.7 substances per case. Lidocaine was the predominant finding followed by caffeine, and diphenhydramine all of which could result from non-illicit drug exposure, however, there were positives for levamisole, phenacetin, noxiptillin and xylazine none of which are approved in the United States for human therapeutic use. This initial set of data established a preliminary positivity rate of potentially toxic adulterants in UCT samples positive for cocaine or opioid use.