Variability of Isoquinoline Alkaloid Profiles and Anticholinesterase Activities with Binding-Mode Predictions of Glaucium flavum Population.

In this study, phytochemical and biological activity studies supported by docking were carried out on a species of the genus Glaucium, a repository of isoquinoline alkaloids. The GC-MS (Gas Chromatography-Mass Spectrometry) method is used to characterize the isoquinoline alkaloids of Glaucium flavum Crantz. (Papaveraceae). G. flavum was collected from seven different regions of Türkiye (Antalya, Urla-Izmir, Mordogan-Izmir, Mugla, Assos-Canakkale, Karabiga-Canakkale, Giresun) and totally 17 compounds were detected by GC-MS. Glaucine was found to be the major constituent in the sample collected from Mugla, whereas isocorydine was recorded to be the principal alkaloid in other samples. Further fractionation studies on G. flavum collected from Antalya province in Southwestern Türkiye, yielded five major alkaloids (isocorydine 1, dihydrosanguinarine 2, glaucine 3, dehydroglaucine 4, protopine 5) which were characterized by spectroscopic methods. Anticholinesterase activities of the extracts and isolated alkaloids were also tested by in vitro Ellman method. The isolated compounds were also analyzed by a molecular docking technique to determine the binding orientations in the gorge of the active site of acetylcholinesterase (AChE) and a homology model of butyrylcholinesterase (BuChE). This is the first comparative investigation of the phytochemical composition and biodiversity of Glaucium flavum species growing in Türkiye.

Assessment of Toxic Pyrrolizidine and Tropane Alkaloids in Herbal Teas and Culinary Herbs Using LC-Q-ToF/MS.

Pyrrolizidine alkaloids are secondary metabolites produced by plants as a defense against insects. These can cause acute or chronic toxicity in humans. Therefore, avoiding potential poisoning from the consumption of tea and culinary plants contaminated with pyrrolizidine alkaloids (PAs), pyrrolizidine alkaloids N-oxides (PANOs), and tropane alkaloids (TAs) is important for human health and food safety. Therefore, it is important to determine the levels of these substances with reliable and highly accurate methods. In this study, the PAs, PANOs, and TAs in herbal teas and culinary herbs sold in Turkish markets were identified and their levels were determined. Thus, the general profiles of herbal teas and culinary herbs in Turkey were revealed, and the compliance of the total amounts of PA and TA with the regulations was examined. The identification and quantification of 25 PAs and N-oxides and 2 TAs (atropine and scopolamine) in the samples was performed with a liquid chromatography-quadrupole time-of-flight tandem mass spectrometer (LC-Q-ToF/MS). At least a few of these substances were detected in all of the tested herbal teas and culinary herbs. The total contents of the black tea, green tea, mixed tea, flavored tea, chamomile tea, sage tea, linden tea, fennel tea, rosehip tea, peppermint, and thyme samples ranged from 4.6 ng g-1 to 1054.5 ng g-1. The results obtained shed light on the importance of analyzing the total dehydro PA, PANO, and TA amounts in plant-based products consumed in diets with sensitive and accurate methods, and they highlight the necessity of performing these analyses routinely in terms of food safety.

Metal-organic framework functionalized with deep eutectic solvent for solid-phase extraction of Rhodamine 6G in water and cosmetic products.

An NH2 -MIL-53(Al)-DES(ChCl-Urea) nanocomposite was synthesized for extraction and determination of Rhodamine (Rh) 6G from environmental and cosmetic samples. The deep eutectic solvent (DES) was prepared by mixing choline chloride and urea in a mole ratio of 1:2. NH2 -MIL-53(Al)-DES(ChCl-Urea) nanocomposite was synthesized using the impregnation method at a ratio of 60:40 (w/w). The optimum conditions were determined after NH2 -MIL-53(Al)-DES(ChCl-Urea) characterization was performed. The optimum conditions were determined as pH 8, adsorbent amount of 15 mg, total adsorption-desorption time of 6 min, and enrichment factor of 20. The recovery values of the solid-phase extraction method for water and cosmetic samples under optimum conditions were between 95% and 106%. NH2 -MIL-53(Al)-DES(ChCl-Urea) nanocomposite was an economically advantageous adsorbent because of its reusability of 15 times. All analyses were performed using the ultraviolet-visible spectrophotometer. The linear range, limit of detection, and limit of quantification of the method were 100-1000, 9.80, and 32.68 µg/L, respectively. The obtained results showed that the synthesized nanocomposite is a suitable adsorbent for the determination of Rh 6G in water and cosmetic samples. The real sample applications were verified with the high-performance liquid chromatography system.

Fe3O4-SiO2-MIL-53 (Fe) nanocomposite for magnetic dispersive micro-solid phase extraction of cadmium (II) at trace levels prior to HR-CS-FAAS detection.

A magnetic metal-organic framework Fe3O4-SiO2-MIL-53 (Fe) nanocomposite was synthesized for magnetic dispersion micro-solid phase extraction (M-d-µSPE) of cadmium in water, spice, chocolate, tea, and tobacco samples prior to the detection by flame atomic absorption spectrometry. Fe3O4-SiO2-MIL-53 (Fe) nanocomposite fabricated using the solvothermal technique was characterized using a field emission scanning electron microscope and X-ray diffraction. The extraction efficiency of the method was improved by optimizing the experimental factors. After optimization, the linearity range for Cd (II) was 4.3-500 µgL-1. The limits of detection and quantification were 1.3 and 4.3 µgL-1, respectively. The presented magnetic dispersion-micro solid phase extraction method was applied to Cd (II) analysis in food and some environmental samples.

Cytotoxicity of subcritical water extracts obtained from byproducts generated at commercial pecan shelling operations on cancer cells.

This study examined potential of the extracts obtained from the byproducts generated at commercial pecan nut-shelling operations in cancer treatment. The subcritical water extracts obtained from two varieties, Native and Pawnee, were analyzed for their phenolic contents and compositions. Effects of the extracts on viability and IC50 of the human cell lines representing a broad range of cancer types, cervical, lung, skin, breast, colon and prostate cancers, were investigated. Although the effect of the temperature on the phenolic contents and compositions of the extracts was not statistically significant, the influence of the variety was extensive. The pecan shell extracts were not cytotoxic to the healthy cell line Vero in the concentration range examined. Some of the pecan shell extracts had greater efficay than Doxorubicin, a drug used in cancer chemotherapy, in reducing cancer cell viability. This study is novel and practical implications of the data generated in this study are noteworthy, because this is the first report on the beneficial effects of subcritical water extracts obtained from pecan shelling industry byproducts on a broad range of cancer cell lines. It is likely that the experimental data presented in this study will support and encourage future research on the biological pathways involved in the interactions of the cancer cells and the extracts. The findings of this study will facilitate research on downstream processing and purification of the crude extracts exhibiting high cancer cell cytotoxcity, potentially improving the final product efficacy and lead to commercial applications.

Chemical Composition and Antioxidant Properties of Pecan Shell Water Extracts.

This study examined the chemical composition and antioxidant properties of the extracts obtained from two byproduct streams generated at a commercial pecan nut shelling operation. Byproduct stream F contained more pecan nut meat pieces and packing material than stream S, consisting of mainly hard outer shell pieces. Samples from Native variety nuts were processed using subcritical, sonication aided and microwave heating, using water as a solvent. Ferric reducing capacity (FRAP), Total Phenolic Content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ABTS [2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)] assays were used to determine antioxidant properties of the extracts. The experimental results clearly demonstrated that the chemical composition of the industrial byproducts was significantly different from the hand-separated shells. All the water extracts exhibited significant DPPH, ABTS and FRAP activity. The highest antioxidant capacity was obtained with the extracts obtained via subcritical water at 80 °C. This is the first report published in the literature on the antioxidant properties of water extracts obtained from industrial byproducts from a pecan nut shelling operation processing Native variety. New data generated in this study expand our knowledge of the properties of industrial nut shelling industry byproducts and help to evaluate the potential use of the shell extracts as antioxidants in various applications.

Sensitive and selective determination of imidacloprid with magnetic molecularly imprinted polymer by using LC/Q-TOF/MS.

In this paper, magnetic-molecularly imprinted polymer was used for the preconcentration of trace levels of imidacloprid in water and apple samples prior to liquid chromatography-quadrupole-time-of-flight mass spectrometric determination. The selectivity of the magnetic polymer was united with the sensitivity and the high resolving power of the chromatographic system. The developed method showed a linear range from 10.0 to 500.0 µg/L. The quantitative recoveries were obtained for water and apple samples in the range of 92.0%-99.0 %. The relative standard deviations of intra-day and inter-day tests were found to be in the range of 0.8%-1.2% and 1.2%-1.6 %, respectively. In addition, the same magnetic-molecularly imprinted polymer (MMIP) can be used at least ten cycles for the determination of imidacloprid. The preconcentration factor of the method was found to be 2.5, and the total preconcentration procedure can be completed in 1 h. Characterization of synthesised particles were executed with various techniques. Due to its suitable limit of detection, dynamic linear range, sensitivity and selectivity, the developed method seemed to be ideal for the determination and preconcentration of imidacloprid in water and fruit samples.

Preparation of glutathione loaded nanoemulsions and testing of hepatoprotective activity on THLE-2 cells.

To improve bioavailability and stability of hydrophobic and hydrophilic compounds, nanoemulsions are good alternatives as delivery systems because of their nontoxic and nonirritant nature. Glutathione (GSH) suffers from low stability in water, where its encapsulation in nanoemulsions is a powerful strategy to its stability in aqueous systems. The aim of this study was to obtain nanoemulsions from the hydrophobic/hydrophilic contents of N. sativa seed oil so as to improve GSH stability along with bioavailability of N. sativa seed oil. Then, the prepared nanoemulsions were tested for in vitro hepatoprotective activity against ethanol toxicity. To the best of our knowledge, there is no study on the test of nanoemulsions by the combination of Nigella sativa seed oils and GSH in hepatoprotective activity. Here, nanoemulsions with different contents were prepared using Nigella sativa seed oils. Content analyses and characterisation studies of prepared nanoemulsions were carried out. In order to investigate the protective effects against to ethanol exposure, THLE-2 cells were pretreated with nanoemulsions for 2 h with the maximum benign dose (0.5 mg/mL of nanoemulsions). Ethanol (400 mM) was introduced to pretreated cells and nontreated cells for 48- or 72-h periods, followed by cell viability assay was carried out. Fluorescence microscopy tests revealed the introduction of the nanoemulsions into THLE-2 cells. The findings show that nanoformulations have promising in vitro hepatoprotective effects on the THLE-2 cell line against ethanol exposure.

An examination of carbopol hydrogel/organogel bigels of thymoquinone prepared by microwave irradiation method.

Nigella sativa L. is shown wide spread over the world and contains many useful phytochemicals. Much of the biological activity of the seeds has been shown due to the presence of thymoquinone (TQ). Its poor aqueous solubility of TQ hinders its delivery to target site. The aim of this work was to prepare TQ bigels composed of Carbopol 974 P NF (C974) in PEG 400 (organogel) or C974 in water (hydrogel) with microwave heating method. A novel technique, high speed homogenization followed by microwave heating, was used to prepare organogels. The pH, electrical conductivity, differential scanning calorimetry, rheological properties, and morphological structure of the formulations have been evaluated, and the effect of microwave on drug content and TQ antioxidant activity has been investigated. The bigels of TQ were successfully produced via high-speed homogenization followed by microwave-assisted heating for the first time in this study. Highly lipophilic TQ was successfully dissolved in organogel, and it was not affected from the microwaves. It can be stated that microwave heating is a promising method to obtain C974 organogels and thus bigels with appropriate above indicated investigated physicochemical characteristics. The time and energy consumption could be decreased with microwave-assisted heating, especially for gel preparation in the field of pharmaceuticals.

Screen printed electrode-based biosensor functionalized with magnetic cobalt/single-chain antibody fragments for cocaine biosensing in different matrices.

On-site detection of substance abuse is an important approach in the preventive and intervention protocols implementations. It is known that the traditional methods are heavy, time-consuming, and need a high level of logistical requirements. As such, biosensors represent great potential to simplify and improve substance abuse detection. In this study, we have designed a functionalized screen-printed electrode (SPE) electrochemical biosensor with cobalt oxide nanoparticles and single-chain antibody fragments (scFvs) for cocaine detection. Different electrochemical techniques such as differential pulse voltammetry, cyclic voltammetry, and electrochemical impedance spectrometry were used to examine the functionality of the designed biosensor. Furthermore, SEM observations were performed to observe the surface changes after functionalization. The results showed that the linearity ranged between 5.0 and 250 ng/mL and a detection limit of 3.6 ng/mL (n = 6). These results were compared to results obtained from Q-TOF/MS where four different matrices (serum, sweat, urine, and saliva) were spiked with 100 ng/mL cocaine and were analyzed by both methods (Biosensor and Q-TOF/MS). Results showed a higher performance of the biosensor compared to traditional methods. In addition, the selectivity of the biosensor was shown in the presence of different interferents where the designed platform showed a specific response to only cocaine. In conclusion, the designed biosensor proposes great potential for portable and on-site substance abuse detection in addition to boasting the capability of reuse of the SPE and thus, reducing the costs related to such applications.

Application of Biofunctionalized Magnetic Nanoparticles Based-Sensing in Abused Drugs Diagnostics.

Real-time detection of substance use is an approach of high interest leading to the optimization of behavioral interventions and drug abuse intervention. The current methods in use suffer many limitations and need high logistical and laboratory requirements. Biosensors have shown a great potential in overcoming these limitations. In the present study, the electrochemical biosensor composed of a screen-printed electrode (SPE) was designed for the detection of synthetic cannabinoid (SC). Antibody-immobilized magnetic nanoparticles were also used to create a surface on the transducer with magnetic interactions in order to detect JWH-073 as a SC model. The use of immobilized magnetic nanoparticles to create working surfaces makes the electrode a reusable SPE which can be reutilized after the cleansing. To examine and observe any possible changes on the surface due to its interaction with the analyte, different electrochemical techniques such as differential pulse voltammetry, cyclic voltammetry, and electrochemical impedance spectrometry were applied. Based on the obtained results, the linearity of the biosensor was found between 5 and 400 ng/mL, and the detection limit was calculated as 22 ng/mL (n = 6) using the 3 Sb/m formula. The biosensor functionality was studied in the presence of some related interferents that showed lower responses than JWH-073, thus demonstrating the good selectivity of the prepared biosensor. Finally, the sensory platform was used to test synthetic urine sample, and the results were compared with obtained results from liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF/MS), which showed that the proposed method could be utilized to identify abuse drugs.

Development of rapid, sensitive and selective fluorimetric method for determination of 1-naphthalene acetic acid in cucumber by using magnetite-molecularly imprinted polymer.

In this study, a novel method based on the determination of 1-naphthalene acetic acid with the usage of magnetite-molecularly imprinted polymer prior to fluorimetric detection has been developed. Magnetite-molecularly imprinted polymer has been used for the first time as selective adsorbent for the determination of 1-naphthalene acetic acid. The adsorption capacity of the synthesized polymer was found to be 2.18 ±â€¯0.36 mg g-1 (n = 3). Limit of detection (LOD) and limit of quantification (LOQ) of the method were found to be 0.75 and 2.50 µg L-1, respectively. Linearity of the calibration graph for the proposed method was observed within the range of 20-700 µg L-1. The proposed method seems to be rapid where the detection procedure for 1-naphthalene acetic acid can be completed within a total time of 1 h. The same imprinted polymer can be used for the determination of 1-naphthalene acetic acid with quantitative sorption and recovery values repeatedly for at least ten times. The effects of some potential organic interferences were investigated. Proposed method has been successfully applied to determine 1-naphthalene acetic acid in cucumber, where the recoveries of the spiked samples were found to be in the range of 93.7-104.5%. Characterization of the synthesized polymer was also evaluated. By combining the high capacity, cheapness, reusability and selectivity of the magnetic adsorbent with the dynamic calibration range, rapidity, simplicity, and sensitivity of fluorimetry, the proposed method seems to be an ideal method for the determination of trace levels of 1-naphthalene acetic acid.

Highly sensitive and selective method for the rapid determination and preconcentration of haloperidol by using a magnetite-molecularly imprinted polymer.

A sensitive and selective method based on the determination of haloperidol with the usage of magnetite-molecularly imprinted polymer and high-resolution liquid chromatography has been developed. This novel method is rapid as the detection procedure for haloperidol can be completed within a total time of 1 h. The same imprinted polymer can be used for the determination of haloperidol at least 20 times. The proposed method has been succesfully applied to synthetic urine and serum samples and the recoveries of the spiked samples were in the range of 94.7-100.7%. The limit of detection and limit of quantification of the method were 2.25 and 7.50 µg/L, respectively. Linearity of the calibration graph was observed within the range of 10-250 µg/L. By combining the high capacity, high selectivity, and reusability of the magnetic adsorbent with the dynamic calibration range, high sensitivity and high resolution of liquid chromatography with quadrupole time-of-flight mass spectrometry, the proposed method is an ideal method for the determination and preconcentration of trace levels of haloperidol. A magnetite-molecularly imprinted polymer has been used for the first time as a selective adsorbent for the determination of haloperidol.

Testing of bioactive-nanovesicles on hepatotoxicity of atypical antipsychotics via digital holography.

Atypical antipsychotic drugs induce hepatic toxicity. Thus, it is of importance to eliminate the side effects of these drugs. Herein we describe the preparation of nanoemulsions with a dietary supplement; wheat germ oil (WGO), to ameliorate the liver damage induced by clozapine and olanzapine. THLE-2 cell line was used as a model to investigate the effects of these nanoemulsions on cell viability as well as antioxidative efficiency after antipsychotic insult. In this context, a conventional cell culture method; MTT was used along with a novel cellular imaging technique called digital holography (DH) to evaluate cell viability. Obtained data confirmed that both clozapine and olanzapine induced the liver damage in in vitro model and WGO nanoemulsions were found to be effective on cells and eliminate the cytotoxic effects of these drugs. Briefly, this study has some outputs as follows; it showed that different dietary supplements can be used in such formulations instead of their pristine forms to increase bioavailability. Also, DH was successfully applied for the monitoring of cell viability and it could be a promising approach as the reactive-free cytotoxicity test.

An aptamer folding-based sensory platform decorated with nanoparticles for simple cocaine testing.

The consumption of illicit drugs such as cannabis, cocaine, and amphetamines is still a major health and social problem, creating an abuse in adults especially. Novel techniques which estimate the drug of abuse are needed for the detection of newly revealed psychoactive drugs. Herein, we have constructed a combinatorial platform by using quantum dots (QDs) and gold nanoparticles (AuNPs) as well as a functional aptamer which selectively recognizes cocaine and its metabolite benzoylecgonine (BE). We have called it an aptamer folding-based sensory device (AFSD). For the fabrication of AFSD, QDs were initially immobilized onto the poly-L-lysine coated µ-well surfaces. Then, the AuNP-aptamer conjugates were bound to the QDs. The addition of cocaine or BE caused a change in the aptamer structure which induced the close interaction of AuNPs with the QDs. Hence, quenching of the fluorescence of QDs was observed depending on the analyte amount. The linearity of cocaine and BE was 1.0-10 nM and 1.0-25 µM, respectively. Moreover, the limits of detection for cocaine and BE were calculated as 0.138 nM and 1.66 µM. The selectivity was tested by using different interfering substances (methamphetamine, bovine serum albumin, codeine, and 3-acetamidophenol). To investigate the use of AFSD in artificial urine matrix, cocaine/BE spiked samples were applied. Also, confirmatory analyses by using high performance liquid chromatography were performed. It is shown that AFSD has a good potential for testing the cocaine abuse and can be easily adapted for detection of various addictive drugs by changing the aptamer according to desired analytes. Copyright © 2016 John Wiley & Sons, Ltd.

Bioconjugation and Applications of Amino Functional Fluorescence Polymers.

Synthesis and novel applications of biofunctional polymers for diagnosis and therapy are promising area involving various research domains. Herein, three fluorescent polymers, poly(p-phenylene-co-thiophene), poly(p-phenylene), and polythiophene with amino groups (PPT-NH2 , PPP-NH2 , and PT-NH2 , respectively) are synthesized and investigated for cancer cell targeted imaging, drug delivery, and radiotherapy. Polymers are conjugated to anti-HER2 antibody for targeted imaging studies in nontoxic concentrations. Three cell lines (A549, Vero, and HeLa) with different expression levels of HER2 are used. In a model of HER2 expressing cell line (A549), radiotherapy experiments are carried out and results show that all three polymers increase the efficacy of radiotherapy. This effect is even more increased when conjugated to anti-HER2. In the second part of this work, one of the selected polymers (PT-NH2 ) is conjugated with a drug model; methotrexate via pH responsive hydrazone linkage and a drug carrier property of PT-NH2 is demonstrated on neuroblastoma (SH-SY5Y) cell model. Our results indicate that, PPT-NH2 , PPP-NH2 , and PT-NH2 have a great potential as biomaterials for various bioapplications in cancer research.