Myricetin induces apoptosis and autophagy by inhibiting PI3K/Akt/mTOR signalling in human colon cancer cells.

BACKGROUND: The compound 3,3',4',5,5',7-hexahydroxyflavone (myricetin) is a natural flavonoid with antitumour activity. Most of the studies on myricetin have focused on the induction of tumour cell apoptosis, and little is known about the regulatory effects of myricetin on autophagy in colorectal cancer. METHODS: Here, we studied the effects of myricetin on colon cancer cell proliferation, apoptosis and autophagy. We detected colon cancer cell apoptosis induced by myricetin via flow cytometry and Hoechst 33258 staining. Transmission electron microscopy was performed to observe the morphological changes associated with autophagy. The expression levels of apoptosis-, autophagy- and PI3K/Akt/mTOR signalling-related proteins were measured by Western blot analysis. RESULTS: This study confirmed that myricetin inhibits the proliferation of 4 kinds of colon cancer cell lines. Myricetin induced cell apoptosis and autophagy by inhibiting PI3K/Akt/mTOR signalling pathway. In addition, the inhibition of autophagy with 3-methyladenine (3-MA) promoted the apoptosis of myricetin-treated colon cancer cells. CONCLUSIONS: Considering that myricetin induces apoptosis and autophagy in colon cancer cells, myricetin may become a viable candidate for chemotherapy; it could be used to exert tumour inhibitory effects alone or as adjuvant chemotherapy to inhibit autophagy. These studies may provide further evidence for the potential use of myricetin in the treatment of colon cancer.

Simultaneous determination of 11 phthalate esters in bottled beverages by graphene oxide coated hollow fiber membrane extraction coupled with supercritical fluid chromatography.

Phthalate esters (PAEs) are a group of serious environmental pollutants, which lead to carcinogenicity or tumorigenicity in human body. In this study, a rapid, sensitive and green method by graphene oxide coated hollow fiber membrane extraction (GO-HFME) coupled with supercritical fluid chromatography (SFC) was proposed for the determination of 11 phthalate esters in bottled beverages. Graphene oxide (GO) was prepared and coated onto a porous hollow fiber membrane (HFM) to reinforce the efficiency of membrane extraction. The modified hollow fiber membrane was employed for the extraction of phthalate esters from bottled beverages prior to the determination by the supercritical fluid chromatography with UV detection. To achieve the maximum extraction efficiency, several parameters were investigated including GO concentration, extraction time, desorption solution and desorption time. SFC variables including stationary phase, modifier composition and percentage, column temperature, flow rate and backpressure were studied to improve the separation conditions. Under these optimized conditions, all the studied 11 phthalate esters were well separated and simultaneously determined in 7 min by SFC. The performance of the developed method was evaluated. Good linearity was observed (R ≥ 0.999) in the range of 0.02-10.0 µg/mL with limit of detection (LOD, S/N = 3) ranging from 1.5 to 3.0 ng/mL. Recoveries of all the PAEs for the spiked samples were between 92.1% and 99.3% with satisfactory relative standard deviations (RSD) less than 5.9%. The proposed method is time-saving, green, simple and robust, which will be an alternative way to the analysis of PAEs in real samples.

Graphene-coated polymeric anion exchangers for ion chromatography.

Carbonaceous stationary phases have gained much attention for their peculiar selectivity and robustness. Herein we report the fabrication and application of a graphene-coated polymeric stationary phase for anion exchange chromatography. The graphene-coated particles were fabricated by a facile evaporation-reduction method. These hydrophilic particles were proven appropriate substrates for grafting of hyperbranched condensation polymers (HBCPs) to make pellicular anion exchangers. The new phase was characterized by zeta potentials, Fourier transform infrared spectroscopy, thermogravimetry and scanning electron microscope. Frontal displacement chromatography showed that the capacities of the anion exchangers were tuned by both graphene amount and HBCPs layer count. The chromatographic performance of graphene-coated anion exchangers was demonstrated with separation of inorganic anions, organic acids, carbohydrates and amino acids. Good reproducibility was obtained by consecutive injections, indicating high chemical stability of the coating.

Poly (styrene-divinyl benzene-glycidylmethacrylate) stationary phase grafted with poly amidoamine (PAMAM) dendrimers for rapid determination of phenylene diamine isomers in HPLC.

A novel high performance liquid stationary phase was prepared by grafting poly amidoamine (PAMAM) dendrimers to poly (styrene-divinyl benzene-glycidylmethacrylate) matrix in this work. Different generations of PAMAM dendrimers grafted to polymer matrices containing different glycidylmethacrylate amounts by reacting with epoxy groups in glycidylmethacrylate molecules. The synthesized PAMAM dendrimers and stationary phase were characterized by fourier transform infrared spectroscopy, scanning electron microscope, N2 adsorption/desorption, elemental analysis and thermo gravimetric analyzer. The proposed stationary phase was suitable for rapid separation of phenylene diamine isomers and aminobenzene. The separation mechanism of benzene rings attraction and amino repulsion was verified by retention behaviors of methylamine, phenylamine, phenylene diamine, naphthylamine and toluene. The reliability of the stationary phase was demonstrated by separation and determination of phenylene diamine isomers including m-phenylene diamine, o-phenylene diamine and p-phenylene diamine in hair dye precursors within 10min. Satisfactory resolution and short analysis time make the columns promising for determination of aniline compounds in various fields.

Detection of trace fluoride in serum and urine by online membrane-based distillation coupled with ion chromatography.

An online membrane-based distillation (MBD) coupled with ion chromatography (IC) method was proposed for automatic detection of trace fluoride (F-) in serum and urine samples. The system consisted of a sample vessel, a lab-made membrane module and an ion chromatograph. Hydrophobic polytetrafluoroethylene (PTFE) hollow fiber membrane was used in MBD which was directly performed in serum and urine samples to eliminate the matrix interferences and enrich fluoride, while enabling automation. The determination of fluoride in biological samples was carried out by IC with suppressed conductometric detection. The proposed method feasibly determined trace fluoride in serum and urine matrices with the optimized parameters, such as acid concentration, distillation temperature, and distillation time, etc. Fluoride exhibited satisfactory linearity in the range of 0.01-5.0mg/L with a correlation coefficient of 0.9992. The limit of detection (LOD, S/N=3) and limit of quantification (LOQ, S/N=10) were 0.78µg/L and 2.61µg/L, respectively. The relative standard deviations of peak area and peak height were all less than 5.15%. The developed method was validated for the determination of fluoride in serum and urine with good spiked recoveries ranging between 97.1-101.9%. This method also can be proposed as a suitable alternative for the analysis of fluoride in other complex biological samples.

Hydrothermal carbonaceous sphere based stationary phase for anion exchange chromatography.

Monodisperse carbonaceous spheres produced by the hydrothermal carbonization of sucrose were first applied as green stationary phase for ion chromatography after quaternization. Depending on the polycondensation of methylamine and 1,4-butanediol diglycidyl ether, polymer containing quaternary ammonium groups were facilely grafted onto the surfaces of hydrothermal carbonaceous spheres (HCSs). The quaternized HCSs with different number of polyelectrolyte layers were characterized by scanning electron microscopy, brunauer-emmett-teller, fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, thermogravimetric analysis and elemental analysis. The measurements of breakthrough curves demonstrated that more layers of grafted polyelectrolyte resulted in higher anion exchange capacity of stationary phase. With good stability, common inorganic anions, monocarboxylic acids, polarizable anions and carbohydrates were effectively separated on the stationary phases, respectively. The high hydrophilicity of HCS surface afforded excellent peak symmetry for all analytes. Furthermore, high-capacity HCSs stationary phase was successfully applied to detect fluoride in tea samples.

Octadecylamine-modified poly (glycidylmethacrylate-divinylbenzene) stationary phase for HPLC determination of N-nitrosamines.

Poly (glycidylmethacrylate-divinylbenzene) (Poly (GMA-DVB)) microspheres were prepared by the two-staged swelling and polymerization method, and modified with octadecylamine (ODA) to obtain ODA-poly (GMA-DVB) stationary phase for HPLC. The new material was characterized by scanning electron microscope, nitrogen adsorption-desorption measurement, Fourier transform infrared spectrum, elemental analysis and thermogravimetric analysis. The results showed that poly (GMA-DVB) microspheres had good monodispersity, porosity and ball shapes. The diameters and specific surface area of the microspheres were about 6µm and 396m(2)g(-1), respectively. ODA-poly (GMA-DVB) stationary phase had good thermal stability. Furthermore, the chromatographic performance of the stationary phase was illustrated by separating n-alkylbenzenes, mono-substituted benzenes and N-nitrosamines. Auxiliary quantum chemistry calculation was also carried out to evaluate the interaction mechanism. According to the evaluation, ODA-poly (GMA-DVB) stationary phase exhibited good hydrophobicity and hydrophobic selectivity, strong stereo-selectivity, polar interaction and π-π interaction. The multi-interaction mechanisms could very likely guarantee its excellent chromatographic performance for the analysis of complex samples. Finally, the column was successfully applied in the determination of N-nitrosamines in pickles sample.

High-capacity anion exchangers based on poly (glycidylmethacrylate-divinylbenzene) microspheres for ion chromatography.

Poly (glycidylmethacrylate-divinylbenzene) microspheres were prepared by the two-staged swelling and polymerization method and applied to prepare anion exchange stationary phases. Methylamine, dimethylamine, trimethylamine, diethylamine and triethylamine were selected to prepare the quaternary ammonium groups of anion exchangers, respectively. The diameters and surface characteristics of microspheres were measured by scanning electron microscope and nitrogen adsorption-desorption measurements. The anion exchangers were characterized by Fourier transform infrared spectrum, elemental analysis and breakthrough curve methods. The chromatographic performances of anion exchangers were illustrated by separating conventional anions, organic weak acids and carbohydrates. The results indicated that the anion exchange capacities were controllable by changing either the content of glycidylmethacrylate in microspheres or the number of bonded quaternary ammonium layer. Meanwhile, the substituents of quaternary ammonium groups greatly influenced the separation properties of anion exchangers. Finally, the three-layer methylamine-quaternized anion exchanger was successfully applied for the determination of fluoride in tea sample. The content of fluoride was detected to be 0.13mgg(-1) without the interference of acetate and formate.

Polystyrene-divinylbenzene-glycidyl methacrylate stationary phase grafted with poly (amidoamine) dendrimers for ion chromatography.

In this work, a novel ion exchange stationary phase based on different generations of poly (amidoamine) dendrimers (PAMAM) was developed for the determination of inorganic anions and carbohydrates. Synthesis of the PAMAM was carried out with the polymerization reaction of ethylenediamine and methyl acrylate. The synthesized PAMAM was then grafted to the polystyrene-divinylbenzene-glycidyl methacrylate (PS-GMA) to form PAMAM-based beads. These beads were finally modified with 1,4-butanediol diglycidyl ether (BDDE) to generate the anion exchanger, which were characterized by scanning electron microscopy (SEM), brunauer-emmett-teller (BET), fourier transform infrared spectroscopy (FTIR), and elemental analysis. Elemental analysis, breakthrough curves and capacity factors showed that more epoxy groups and higher PAMAM generations in stationary phase could result in higher anion exchange capacity. The efficiency, durability and stability of the proposed anion exchanger were investigated by using six inorganic anions (fluoride, chloride, nitrite, bromide, nitrate and sulfate) and four carbohydrates (trehalose, glucose, maltotriose and galacturonic acid) as analytes, respectively. The reliability of the proposed ion chromatographic stationary phase was demonstrated by determining the content of galacturonic acid in polysaccharides from Poria cocos and Atractylodes macrocephala. The relative standard deviations of retention time, peak height, and peak area for galacturonic acid were 0.39%, 1.22%, and 2.02%, respectively. The spiked recoveries were in the range of 88.29%-100.51% for plant polysaccharides. Due to the good structural homogeneity, intense internal porosity, biological compatibility and high density of active groups in PAMAM, this grafted stationary phase showed good ion-exchange characteristics, especially in biological charged molecules.

Preparation of poly(glycidylmethacrylate-divinylbenzene) weak acid cation exchange stationary phases with succinic anhydride, phthalic anhydride, and maleic anhydride for ion chromatography.

In this work, poly(glycidylmethacrylate-divinylbenzene) microspheres were prepared and applied for the preparation of weak acid cation exchange stationary phases. Succinic anhydride, phthalic anhydride, and maleic anhydride were selected as carboxylation reagents to prepare three weak acid cation exchangers by direct chemical derivatization reaction without solvent or catalyst. The diameters and dispersity of the microspheres were characterized by scanning electron microscopy; the amount of accessible epoxy groups and mechanical stability were also measured. The weak acid cation exchangers were characterized by Fourier transform infrared spectroscopy; the content of carboxyl groups was measured by traditional acid base titration method. The chromatographic properties were characterized and compared by separating alkali, alkaline earth metal ions and ammonium and polar amines. The separation properties enhanced in the order of succinic anhydride, phthalic anhydride, and maleic anhydride modified poly(glycidylmethacrylate-divinylbenzene) cation exchangers.

Analysis of sucrose acetates in a crude 6-O-acetyl sucrose product by on-line hydrolysis-high-performance liquid chromatography with pulsed amperometric detection.

A standard-free and sensitive method was developed for analysis of sucrose acetates in a crude 6-O-acetyl sucrose (S-6-a) product by on-line hydrolysis-high-performance liquid chromatography with pulsed amperometric detection (PAD). Sucrose, three regio-isomers of acetyl sucrose and five regio-isomers of diacetyl sucrose were separated on a C18 column using 3% (v/v) acetonitrile in water as eluent within 25min. After purification with LC followed by semi-preparative HPLC, their chemical structures were identified by 1D, 2D NMR and LC-MS. Moreover, quantification of those regio-isomers was achieved by on-line alkaline hydrolysis to liberate sucrose using a post-column delivery system, and then detected by PAD for indirect estimation of the sucrose acetate content. Under optimal conditions, the linear ranges were from 0.03 to 150µmolL(-1) for sucrose corresponding to sucrose acetates with coefficient of determination as 0.9997 and detection limit as 0.01µmolL(-1) (S/N=3). Good repeatability was obtained (RSD<3%, n=6). Furthermore, this method has been successfully applied to the analysis of sucrose and sucrose acetates in a crude S-6-a product during synthesis, purification and structure elucidation studies. The recoveries were from 94.89% to 102.31% for sucrose and sucrose acetates.

Simultaneous determination of sucralose and related compounds by high-performance liquid chromatography with evaporative light scattering detection.

Sucralose is widely used in food and beverages as sweetener. Current synthesis approaches typically provide sucralose products with varying levels of related chlorinated carbohydrates which can affect the taste and flavor-modifying properties of sucralose. Quantification of related compounds in sucralose is often hampered by the lack of commercially available standards. In this work, nine related compounds were purified (purity>97%) and identified by liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR), then a rapid and simple HPLC coupled with evaporative light scattering detection (ELSD) method has been developed for the simultaneous determination of sucralose and related compounds. Under optimized conditions, the method showed good linearity in the range of 2-600µgmL(-1) with determination coefficients R(2)⩾0.9990. Moreover, low limits of detection in the range of 0.5-2.0µgmL(-1) and good repeatability (RSD<3%, n=6) were obtained. Recoveries were from 96.8% to 101.2%. Finally, the method has been successfully applied to sucralose quality control and purification process monitoring.

Fabrication of electrolytic cell for online post-column electrochemical derivatization in ion chromatography.

An electrolytic cell (EC), composed of two ruthenium-plated titanium electrodes separated by cation-exchange membranes, was fabricated and evaluated for online postcolumn derivatization in ion chromatography (IC). Folic acid (FA) and methotrexate (MTX) were preliminarily used as prototype analytes to test the performance of EC. After separation by an anion exchange column, FA and MTX, which emit very weak fluorescence when excited, were electrochemically oxidized online in the anode chamber of the EC. The compounds with strong fluorescence, which are oxidation products, were detected by the fluorescence detector. The phosphate buffer solution (100 mM KH(2)PO(4)) served as an optimal eluent for anion exchange chromatographic separation and a suitable supporting electrolyte for electro-oxidation, leading to ideal compatibility between IC separation and the postcolumn electrochemical derivatization. For the presently proposed method, the linear ranges were from 0.01 mg L(-1) to 5 mg L(-1) for both FA and MTX. The detection limits of FA and MTX were 1.8 and 2.1 µg L(-1), and the relative standard deviations (RSD, n=7) were 2.9% and 3.6%, respectively. The method was applied for the simultaneous determination of FA and MTX in the plasma of patients being treated for rheumatoid arthritis. The determination of MTX in the urine of the patients of diffuse large B cell lymphoma was also demonstrated.

A single pump column-switching technique coupled with polystyrene-divinylbenzene-carbon nanotubes column for the determination of trace anions in different concentrated organic matrices by ion chromatography.

An ion chromatographic method with on-line sample pretreatment was developed for the trace analysis of seven common anions in concentrated matrices. The pretreatment column used in this study consisted of polystyrene-divinylbenzene (PS-DVB) and multi-walled carbon nanotubes (MWCNTs). It was too hydrophobic to retain different inorganic anions, but it showed a strong affinity for organic compounds. Thus, this chromatographic system could be used to detect trace anions in organic solvents, organic acids and relevant salts. The addition of MWCNTs decreased the surface areas of stationary phases and the retention times of organic matrices were shortened. Compared with conventional column-switching technique, only a single instrument (ICS2100) was needed in this system, including a pump, a conductivity detector, an eluent generator, a six-port valve and a ten-port valve. An electrochemical self-generating suppressor (ESGS) was adopted to convert the eluent of KOH into water for the matrix elimination. Two different eluent were employed in the chromatographic system, one for separation and the other for matrix elimination. The sample pretreatment and analysis were realized simultaneously. After optimization of this system, a calibration study was conducted by preparing and analyzing eight concentrations (between 5 and 5000 µg L(-1)) of mixture standards of seven anions in deionized water. The linearity was between 0.9990 and 0.9998, and the detection limits ranged from 0.41 to 3.17 µg L(-1). A spiking study was performed on three representative organic chemicals with satisfactory recoveries between 88.1% and 118.5% when the concentrations of the matrices did not exceed 10 g L(-1).

Preparation, characterization, and analytical applications of a novel polymer stationary phase with embedded or grafted carbon fibers.

A polymer-based chromatographic stationary phase with embedded or grafted multi-walled carbon nanotubes (MWCNTs) has been developed. Three different synthetic methods were utilized to combine the nano-fibers with the substrate of polystyrene-divinylbenzene (PS-DVB). After optimizing the synthetic conditions, this novel polystyrene-divinylbenzene-carbon nanotube (PS-DVB-CNT) stationary phase was characterized by scanning electron microscopy, Raman spectroscopy, thermogravimetric analysis, chemical adsorption and desorption measurement, and mechanical stability test. Compared to PS-DVB particles, PS-DVB-CNT particles have certain improvement in physical and chromatographic performances because the addition of MWCNTs has altered the structures of the particles. The novel stationary phase owns satisfactory resolution, wide pH endurance, and long lifetime, which can be used as an extent to normal HPLC.