Development and validation of an HPLC method to determine metabolites of 5‐hydroxymethylfurfural (5‐HMF)

The food component 5‐hydroxymethylfurfural is supposed to have antioxidative properties and is therefore used as an acting agent in a novel anticancer infusion solution, named Karal®, and an oral supplementation. Previous studies showed that after oral and intravenous application, the substance is completely decomposed to its metabolites: 5‐hydroxymethylfuroic acid, 2,5‐furandicarboxylic acid, and N‐(hydroxymethyl)furoyl glycine. The formation of a fourth metabolite, namely 5‐sulphoxymethylfurfural, is still not clarified according to literature. Due to commercial unavailability, synthesis of 5‐sulphoxymethylfurfural was conducted and a synthesis procedure for N‐(hydroxymethyl)furoyl glycine had to be developed. Identification of the synthesised compounds was proven by LC‐MS and NMR. An appropriate HPLC method was established to obtain good separation of the four possible metabolic substances and 5‐hydroxymethylfurfural within 12 min via a HILIC column (150 × 4.6 mm, 5 μm) using a gradient grade system switching from mobile phase A (ACN/ammonium formate 100 mM, pH 2.35, 95:5, v/v) to mobile phase B (ACN/ammonium formate 100 mM, pH 2.35, 85:15, v/v). The procedure was afterward validated following ICH guidelines in terms of selectivity, linearity, precision, LOD, and LOQ.     

Synthesis and characterization of monosubstituted and disubstituted poly(3,4‐propylenedioxythiophene) derivatives with high electrochromic contrast in the visible region

Monosubstituted and disubstituted 3,4‐propylenedioxythiophenes were synthesized and polymerized by both chemical and electrochemical methods. All the monomers were characterized for their molecular structures, and the polymers were characterized for their electrochemical properties. The disubstituted derivatives showed higher contrast than the corresponding monoalkyl derivatives. The highest electrochromic contrast of 89% was exhibited by a dibenzyl derivative, but the derivative was insoluble. On the other hand, the electrochemically polymerized dihexyl‐ and didodecyl‐substituted poly(3,4‐propylenedioxythiophene)s exhibited 74 and 77% electrochromic contrast, respectively, and were soluble. The molecular weights of the chemically and electrochemically synthesized polymers were analyzed by gel permeation chromatography. The chemically synthesized polymers showed higher molecular weights. © 2004 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 43: 419–428, 2005     

Pseudohalide‐directed Assembly of Two Zinc(II) Coordination Polymers with a 3,4‐Bis(3‐pyridyl)‐5‐(4‐pyridyl)‐1,2,4‐triazole Tecton

The zinc(II) pseudohalide complexes {[Zn(L³³⁴)(SCN)₂(H₂O)](H₂O)₂}_n ( 1 ) and [Zn(L³³⁴)(dca)₂]_n ( 2 ) were synthesized and characterized using the ligand 3,4‐bis(3‐pyridyl)‐5‐(4‐pyridyl)‐1,2,4‐triazole (L³³⁴) and ZnCl₂ in presence of thiocyanate (SCN^–) and dicynamide [dca, N(CN)₂^–] respectively. Single‐crystal X‐ray structural analysis revealed that the central Zn^II atoms in both complexes have similar octahedral arrangement. Compound 1 has a 2D sheet structure bridged by bidentate L³³⁴ and double μ_N,S‐thiocyanate anions, whereas complex 2 , incorporating with two monodentate dicynamide anions, displays a two‐dimensional coordination framework bridged by tetradentate L³³⁴ ligand. Structural analysis demonstrated that the influence of pseudohalide anions plays an important role in determining the resultant structure. Both complexes were characterized by IR spectroscopy, microanalysis, and powder X‐ray diffraction techniques. In addition, the solid fluorescence and thermal stability properties of both complexes were investigated.     

One—Pot Synthese of 3,4—Dihydropyrimidine—2（1H）—thiones Catalyzed by La（OTf）3

A general and practical procedure for the syntheses of 3,4-di-hydropyrimidine-2(1H)-thiones by a one-pot condensation of aldehyde,β-ketoester or β-diketone and thiourea using La(OTf)3 as the catalyst is described.Mild reaction conditions,excellent yields as well as the environmentally friendly character of La(OTf)3 make it an important alternative to the classic acid-catalyzed Biginelli‘s reaction.     

Controlling the Poly(ε‐caprolactone) Degradation to Maintain the Stemness and Function of Adipose‐Derived Mesenchymal Stem Cells in Vascular Regeneration Application

Biodegradable poly(ε‐caprolactone) (PCL) scaffolds with adipose‐derived mesenchymal stem cells (ADSCs) have been used in vascular regeneration studies. An evaluation method of the effect of PCL degradation products (DP) on the viability, stemness, and differentiation capacities of ADSCs is established. ADSCs are cultured in medium containing different concentrations of PCL DP before evaluating the effect of PCL DP on the cell apoptosis and proliferation, cell surface antigens, adipogenic and osteogenic differentiation capacities, and capacities to differentiate into endothelial cells and smooth muscle cells. The results demonstrate that PCL DP exceed 0.05 mg mL^?1 may change the stemness and differentiation capacities of ADSCs. Therefore, to control the proper concentration of PCL DP is essential for ADSCs in vascular regeneration application.     

Mechanism of Gold(I)‐Catalyzed Conia‐ene Reaction of β‐Ketoesters with Alkynes: A DFT Study

Density functional calculations have been performed to comparatively investigate two possible pathways of Au(I)‐catalyzed Conia‐ene reaction of β‐ketoesters with alkynes. Our studies find that, under the assistance of trifluoromethanesulfonate (TfO), the β‐ketoester is the most likely to undergo Model II to isomerize into its enol form, in which TfO plays a proton transfer role through a 6‐membered ring transition state. The coordination of the Au(I) catalyst to the alkynes triple bond can enhance the eletrophilic capability and reaction activity of the alkynes moiety, which triggers the nucleophilic addition of the enol moiety on the alkynes moiety to give a vinyl‐Au intermediate. This cycloisomerizaion step is exothermal by 21.3 kJ/mol with an energy barrier of 56.0 kJ/mol. In the whole catalytic process, the protonation of vinyl‐Au is almost spontaneous, and the formation of enol is a rate‐limiting step. The generation of enol and the activation of Au(I) catalyst on the alkynes are the key reasons why the Conia‐ene reaction can occur in mild condition. These calculations support that Au(I)‐catalyzed Conia‐ene reactions of β‐ketoesters with alkynes go through the pathway 2 proposed by Toste.     

Structure Investigation of Bridgehead Aziridine: Synthesis,Theoretical, and Crystallographic Study of 2,4,6‐Triphenyl‐1,3‐diazabicyclo[3.1.0]hex‐3‐ene

A one‐pot three‐component procedure to efficiently create the 1,3‐diazabicyclo[3.1.0]hex‐3‐ene system is reported. The molecular structure of 2,4,6‐triphenyl‐1,3‐diazabicyclo[3.1.0]hex‐3‐ene ( 3 ) was studied by X‐ray diffraction and compared to ab initio and density‐functional‐theory (DFT) calculations restricted to the core moiety. Geometry optimizations for structural isomers and tautomeric forms of this aziridine fragment, taken as simplified models, were carried out at high calculation levels. Moreover, the same methods were utilized to evaluate the proton affinity of two crucial aziridine tautomers.     

Electrospun fiber mats of poly(3‐hydroxybutyrate), poly(3‐hydroxybutyrate‐co‐3‐hydroxyvalerate), and their blends

Electrospinning of poly(3‐hydroxybutyrate) (PHB), poly(3‐hydroxybutyrate‐co‐3‐hydroxyvalerate) (PHBV), and their blends was first carried out in chloroform at 50 °C on a stationary collector. The average diameter of the as‐spun fiber from PHB and PHBV solutions decreased with increasing collection distance and increased with increasing solution concentration and applied electrical potential. In all of the spinning conditions investigated, the average diameter of the as‐spun pure fibers ranged between 1.6 and 8.8 μm. Electrospinning of PHB, PHBV, and their blends was carried out further at a fixed solution concentration of 14% w/v on a homemade rotating cylindrical collector. Well‐aligned, cross‐sectionally round fibers without beads were obtained. The average diameter of the as‐spun pure and blend fibers ranged between 2.3 and 4.0 μm. The as‐spun fiber mats appeared to be more hydrophobic than the corresponding films and much improvement in the tensile strength and the elongation at break was observed for the blend fiber mats over those of the pure fiber ones. Lastly, indirect cytotoxicity evaluation of the as‐spun pure and blend fiber mats with mouse fibroblasts (L929) indicated that these mats posed no threat to the cells. © 2006 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 44: 2923–2933, 2006     

Development and validation of an HPLC method for tetracycline‐related USP monographs

A novel reversed‐phase HPLC method was developed and validated for the assay of tetracycline hydrochloride and the limit of 4‐epianhydrotetracycline hydrochloride impurity in tetracycline hydrochloride commercial bulk and pharmaceutical products. The method employed L1 (3 µm, 150 × 4.6 mm) columns, a mobile phase of 0.1% phosphoric acid and acetonitrile at a flow rate of 1.0 mL/min, and detection at 280 nm. The separation was performed in HPLC gradient mode. Forced degradation studies showed that tetracycline eluted as a spectrally pure peak and was well resolved from its degradation products. The fast degradation of tetracycline hydrochloride and 4‐epianhydrotetracycline hydrochloride in solution was retarded by controlling the autosampler temperature at 4 °C and using 0.1% H₃PO₄ as diluent. The robustness of the method was tested starting with the maximum variations allowed in the US Pharmacopeia (USP) general chapter Chromatography <621>. The method was linear over the range 80–120% of the assay concentration (0.1 mg/mL) for tetracycline hydrochloride and 50–150% of the acceptance criteria specified in the individual USP monographs for 4‐epianhydrotetracycline hydrochloride. The limit of quantification for 4‐epianhydrotetracycline hydrochloride was 0.1 µg/mL, 20 times lower than the acceptance criteria. The method was specific, precise, accurate and robust. Copyright © 2014 John Wiley & Sons, Ltd.     

Development and validation of a single HPLC method for determination of α‐tocopherol in cell culture and in human or mouse biological samples

A straightforward and common analytical method for α‐tocopherol (αT) determination in various biological samples, including plasma, red blood cells (RBC), tissues and cultured cell lines, was developed and validated, using a reverse phase‐chromatographic method (RP‐HPLC). Even though many chromatographic methods for αT determination have been reported, most of them require readjustment when applied to different types of samples. Thus, an effective and simple method for αT determination in different biological matrices is still necessary, specifically for translational research. This method was applied using a C₁₈ column (250 × 4.6 mm, 5 µm particle size) under isocratic elution with MeOH:ACN:H₂O (90:9:1 v/v/v) at a flow rate of 1 mL/min and detected using photodiode array at 293 nm. Linearity (r >0.9997) was observed for standard calibration with inter‐ and intraday variation of standard <4%. Lower limits of detection and quantification for αT in this assay were 0.091 and 0.305 µg/mL respectively. Validation proved the method to be selective, linear, accurate and precise. The method was successfully applied in great variety of biological samples, that is, human and mouse plasma, RBCs, murine tissues and human/mouse/rat cultured cell lines. More importantly, a single protocol of extraction and detection can be applied, making this method very convenient for standardization of different types of samples. Copyright © 2014 John Wiley & Sons, Ltd.     

Synthesis of New Furo[3,4‐b]quinolin‐1(3H)‐one Scaffolds Derived from γ‐Lactone‐Fused Quinolin‐4(1H)‐ones

In the context of our aim of discovering new antitumor drugs among synthetic γ‐lactone‐ and γ‐lactam‐fused 1‐methylquinolin‐4(1H)‐ones, we developed a rapid access to 5‐methyl‐1,3‐dioxolo[4,5‐g]furo[3,4‐b]quinoline‐8,9(5H,6H)‐dione ( 9 ) exploiting the γ‐lactone‐fused chloroquinoline 10 previously synthesized in our laboratory (Scheme 1). We also elaborated efficient synthetic methods allowing for a rapid access to two nonclassical bioisosteres of 9 , i.e., a deoxy and a carba analogue. The deoxy analogue 11 was prepared in two steps from the γ‐lactone‐fused quinoline 13 which was also the synthetic precursor of 10 (Scheme 1). The carba analogue 6,9‐dihydro‐5‐methyl‐9‐methylene‐1,3‐dioxolo[4,5‐g]furo[3,4‐b]quinolin‐8(5H)‐one ( 12 ) was easily prepared by HCl elimination from the 9‐(chloromethyl)dioxolofuroquinoline 15 , which was obtained via a three‐component one‐pot reaction from N‐methyl‐3,4‐(methylenedioxy)aniline (=N‐methyl‐1,3‐benzodioxol‐5‐amine; 16 ), commercially available chloroacetaldehyde, and tetronic acid ( 17 ) (Scheme 2).     

HPLC method validation for measurement of sulforaphane level in broccoli by‐products

A simple and specific analytical method was developed and tested for the determination of sulforaphane in broccoli by‐products. The method includes the optimization of the conversion of glucoraphanin to sulforaphane, followed by purification of extracts using solid‐phase extraction and high‐performance liquid chromatography (HPLC) analysis. The response surface methodology was used to find optimum conditions for the preparation and purification procedure. Chromatographic conditions for reversed‐phase HPLC with UV photodiode array detection were as follows: column, Exil ODS C₁₈, 25 × 0.46 cm, 5 μm; column temperature, 36°C; mobile phase, a 30 : 70 (v/v) mixture of acetonitrile:water; flow rate, 0.6 mL/min. The detection wavelength was UV 202 nm. Under these conditions, excellent linearity was obtained (r² = 1), and the overall recovery was 97.5 and 98.1% for fresh florets and lyophilized florets, respectively. The precision results showed that the relative standard deviation of the repeatability for florets fresh and lyophilized was 3.0 and 4.0%, respectively. Sulforaphane contents were determined in the edible portion of fresh broccoli, and broccoli crop remains. Copyright © 2009 John Wiley & Sons, Ltd.     

Synthesis of allyl end‐block functionalized poly(ε‐caprolactone)s and their facile post‐functionalization via thiol–ene reaction

A simple and facile strategy for the functionalization of commercial poly(ε‐caprolactone) diols (PCLs) with pendant functionalities at the polymer chain termini is described. Well‐defined allyl‐functionalized PCLs with varying numbers of allyl end‐block side‐groups were synthesized by cationic ring‐opening polymerization of allyl glycidyl ether using PCL diols as macroinitiators. Further functionalization of the allyl‐functionalized PCLs was realized via the UV‐initiated radical addition of a furan‐functionalized thiol to the pendant allyl functional groups, showing the suitability for post‐modification of the PCL materials. Changes in polymer structure as a result of varying the number of pendant functional units at the PCL chain termini were demonstrated. © 2017 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2017 , 55, 928–939     

Synthesis and Antibacterial Activity of 3‐(5‐Methylisoxazol‐3‐yl) −1,2,4‐triazolo [3,4‐b]‐1,3,4‐thiadiazine Derivatives

The condensed products 2‐10 of 4‐amino‐5‐mercapto‐3‐(5‐methylisoxazol‐3‐yl)‐l,2,4‐triazole (1) with chloroacetaldehyde, 2‐bromocyclohexanone, chloranil, ωbromo‐ω‐(1H‐1, 2,4‐triazol‐l‐yl)acetophenone, 2‐bromo‐4′‐substituted acetophenones and 2‐bromo‐6′‐methoxy‐2′‐acetonaphthone were described. The antibacterial activities were also evaluated.     

Alk‐1‐ene Polymerization in the Presence of a Monocyclopentadienyl Zirconium(IV) Acetamidinate Catalyst: Microstructural and Mechanistic Insights

Suitably activated, (Cp*){N(^tBu)C(Me)N(Et)}ZrMe₂ is known to initiate the ‘living’ and isotactic‐selective polymerization of alk‐1‐enes, and it can be used to synthesize block copolymers and stereoblock polymers. We report a full molecular kinetic investigation of propene, but‐1‐ene, and hex‐1‐ene polymerization with a MAO‐activated catalyst system. By combining NMR microstructural polymer analysis with QM modeling of the active species, the complicated regio‐ and stereochemistry of the polyinsertion process, as well as the active chain‐transfer pathways, are investigated. The perspectives and limitations of this catalyst for application in (stereo)block polymerizations are discussed.

     

Phosphotungstic acid grafted zeolite imidazolate framework as an effective heterogeneous nanocatalyst for the one‐pot solvent‐free synthesis of 3,4‐dihydropyrimidinones

Phosphotungstic acid (H₃PW₁₂O₄₀, PTA) supported on ZIF‐9(NH₂) was synthesized for the first time and performed as an effective and environmental friendly catalyst in the one‐pot three component Biginelli condensation of different substituted benzaldehydes with ethyl acetoacetate and urea to afford the corresponding 3,4‐dihydropyrimidin‐2‐(1H)‐ones under solvent‐free conditions. ZIF‐9(NH₂) and the prepared nanocatalyst PTA@ZIF‐9(NH₂) were characterized by XRD, FESEM, TEM, EDX, BET, AAS, TGA, UV–Vis, and FT‐IR. After reaction, the nanocatalyst can be easily separated from the reaction mixture by centrifuge and the recovered catalyst can be reused for at least five times with a 14% reduction in yield after the fifth run. This study showed that ZIF‐9(NH₂) can be utilized as a promising support for PTA and developed a highly active, stable and reusable heterogeneous catalyst under easy reaction condition in the multi‐component organic synthesis.     

Development of an Efficient Procedure for Determination of Copper,Zinc and Iron after Solid Phase Extraction on 3‐(1‐(1‐H‐Indol‐3‐yl)‐3‐Phenylallyl)‐1H‐Indole Loaded on Duolite XAD 761

A method for the simultaneous preconcentration of Cu²⁺,Zn²⁺ and Fe³⁺ ions, in some food samples has been reported. The method is based on the adsorption of 3‐(1‐(1‐H‐indol‐3‐yl)‐3‐phenylallyl)‐1H‐indole (IPAI) loaded on Duolite XAD 761. The metal ions adsorbed on the modified solid phase resin are eluted using 6 mL of 4 mol L^?1 nitric acid. The influences of the analytical parameters including pH and amount of ligand and solid phase and type and amount of surfactant and sample volume on the metal ions recoveries were investigated. The effects of matrix ions on the retentions of the analytes were also examined. The recoveries of analytes were generally higher than 95% with a RSD lower than 5%. The method has been successfully applied for these metals content evaluation in some real samples.     

Development and validation of a stability‐indicating HPLC method for topiramate using a mixed‐mode column and charged aerosol detector

The analysis of topiramate in the presence of its main degradation products is challenging due to the absence of chromophore moieties and their wide range of polarity. Mixed‐mode chromatography has been used in such cases because it combines two or more modes of separation. Charged aerosol detector is also an alternative since its detection is independent of optical properties and analyte ionization. This study is aimed to develop and validate two new stability‐indicating methods by high‐performance liquid chromatography for the main degradation products of topiramate using mixed‐mode chromatography and a charged aerosol detector. Method 1 employed an Acclaim Trinity P1® column (3.0 mm × 150 mm, 2.7 μm) with a mobile phase comprising of 80% ammonium acetate buffer (20 mM, pH 4.0) and 20% methanol at a flow rate of 0.5 mL/min at 35°C. Method 2 utilized a C18 Acclaim 120® column (4.6 mm × 250 mm; 5 μm) with ACN/water (50:50) at a flow rate of 0.6 mL/min at 50°C. Validation of the two methods demonstrated excellent performance with respect to linearity, precision, accuracy, and selectivity. The limits of detection for topiramate, fructose, sulfate, sulfamate, and compound A were 2.97, 12.08, 4.02, 13.91, and 3.94 μg/mL, respectively.     

A Green Approach for the Synthesis and Thiol‐ene Modification of Alkene Functio1489lized Poly(2‐oxazoline)s

The bulk polymerization of 2‐(dec‐9‐enyl)‐2‐oxazoline ( DecEnOx ), a fatty acid‐based monomer for the cationic ring‐opening polymerization, is reported. Furthermore, under optimal conditions, namely microwave heating at 100 °C, the bulk copolymerization with 2‐ethyl‐2‐oxazoline yielded well‐defined copolymers. Due to its pendant alkene groups DecEnOx ‐based polymers possess the potential to be modified in efficient thiol‐ene reactions. The functionalization with thiols, e.g., dodecanethiol and 2,3,4,6‐tetra‐O‐acetyl‐1‐thio‐β‐D ‐glycopyranose in “green” solvents is demonstrated.

     

The development and validation of a high‐throughput LC–MS/MS method for the analysis of endogenous β‐hydroxy‐β‐methylbutyrate in human plasma

A high‐throughput, sensitive, and rugged liquid chromatography–tandem mass spectrometry (LC–MS/MS) method for the rapid quantitation of β ‐hydroxy‐β ‐methylbutyrate (HMB) in human plasma has been developed and validated for routine use. The method uses 100 μL of plasma sample and employs protein precipitation with 0.1% formic acid in methanol for the extraction of HMB from plasma. Sample extracts were analyzed using LC–MS/MS technique under negative mode electrospray ionization conditions. A ¹³C–labeled stable isotope internal standard was used to achieve accurate quantitation. Multiday validation was conducted for precision, accuracy, linearity, selectivity, matrix effect, dilution integrity (2×), extraction recovery, freeze–thaw sample stability (three cycles), benchtop sample stability (6 h and 50 min), autosampler stability (27 h) and frozen storage sample stability (146 days). Linearity was demonstrated between 10 and 500 ng/mL. Inter‐day accuracies and coefficients of variation (CV) were 91.2–98.1 and 3.7–7.8%, respectively. The validated method was proven to be rugged for routine use to quantify endogenous levels of HMB in human plasma obtained from healthy volunteers.