Development and validation of a sensitive UHPLC‐MS/MS method for the simultaneous analysis of tramadol,dextromethorphan chlorpheniramine and their major metabolites in human plasma in forensic context: application to pharmacokinetics

The prerequisites for forensic confirmatory analysis by LC/MS/MS with respect to European Union guidelines are chromatographic separation, a minimum number of two MS/MS transitions to obtain the required identification points and predefined thresholds for the variability of the relative intensities of the MS/MS transitions (MRM transitions) in samples and reference standards. In the present study, a fast, sensitive and robust method to quantify tramadol, chlorpheniramine, dextromethorphan and their major metabolites, O‐desmethyltramadol, dsmethyl‐chlorpheniramine and dextrophan, respectively, in human plasma using ibuprofen as internal standard (IS) is described. The analytes and the IS were extracted from plasma by a liquid–liquid extraction method using ethyl acetate–diethyl‐ether (1:1). Extracted samples were analyzed by ultra‐high‐performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry (UHPLC‐ESI‐MS/MS). Chromatographic separation was performed by pumping the mobile phase containing acetonitrile, water and formic acid (89.2:11.7:0.1) for 2.0 min at a flow rate of 0.25 μL/min into a Hypersil‐Gold C₁₈ column, 20 × 2.0 mm (1.9 µm) from Thermoscientific, New York, USA. The calibration curve was linear for the six analytes. The intraday precision (RSD) and accuracy (RE) of the method were 3–9.8 and ?1.7–4.5%, respectively. The analytical procedure herein described was used to assess the pharmacokinetics of the analytes in 24 healthy volunteers after a single oral dose containing 50 mg of tramadol hydrochloride, 3 mg chlorpheniramine maleate and 15 mg of dextromethorphan hydrobromide. Copyright © 2014 John Wiley & Sons, Ltd.     

Phenolic content,antioxidant and anti-inflammatory activities of Tunisian Diplotaxis simplex (Brassicaceae)

This study investigates the polyphenol content of Diplotaxis simplex extract and the biological activities of the main organ. The analysed extracts showed that polyphenol contents varied considerably as a function of organs. Furthermore, novel biological activities of this species were assessed. Flower extracts exhibit a potent in vitro antioxidant capacity using oxygen radical absorbance capacity and displayed a strong anti-inflammatory activity, inhibiting nitric oxide release, by 79.3% at 160 μg/mL. Our findings suggested that the Diplotaxis flower is a valuable source of antioxidants and anti-inflammatory agents.     

Ultrasensitive detection and identification of BRAF V600 mutations in fresh frozen,FFPE, and plasma samples of melanoma patients by E-ice-COLD-PCR

A number of molecular diagnostic methods have been developed for the detection and identification of mutations in tumor samples, which are important for the choice of treatment in the context of personalized medicine. For the treatment of metastatic melanoma, Vemurafenib is recommended for patients with BRAF V600 activating mutations. However, the different assays developed to date for the detection of these mutations lack sensitivity or specificity or do not allow a sequencing-based identification or validation of the mutation. Recently, enhanced improved and complete enrichment co-amplification at lower denaturation temperature-polymerase chain reaction (E-ice-COLD-PCR) has been developed as a sensitive method for the detection and identification of mutations in KRAS codons 12/13. Here, we present the first E-ice-COLD-PCR assay for the detection and identification of BRAF codon 600 mutations, which has a large dynamic range, as 25 pg to 25 ng can be used as DNA input without any reduction in mutation enrichment efficiency, and which can detect down to 0.01 % of mutated alleles in a wild-type background. The assay has been validated on fresh frozen, formalin-fixed paraffin-embedded (FFPE), and plasma samples of melanoma patients and has allowed the detection and identification of BRAF mutations present in samples appearing as wild type using standard pyrosequencing, endpoint genotyping, or Sanger sequencing. Thus, the BRAF V600 E-ice-COLD-PCR assay is currently one of the most powerful molecular diagnostic tools for the ultrasensitive detection and identification of BRAF codon 600 mutations.     

Electroswitchable photoluminescence activity: synthesis, spectroscopy, electrochemistry, photophysics, and X-ray crystal and electronic structures of [Re(bpy)(CO)3(C[triple bond]C[bond]C6H4[bond]C[triple bond]C)Fe(C5Me5)(dppe)][PF6](n) (n = 0, 1)

A novel heterobimetallic alkynyl-bridged complex, [Re(bpy)(CO)(3)(C[triple bond]C[bond]C(6)H(4)[bond]C[triple bond]C)Fe(C(5)Me(5))(dppe)], 1, and its oxidized species, [Re(bpy)(CO)(3)(C[triple bond]C[bond]C(6)H(4)[bond]C[triple bond]C)Fe(C(5)Me(5))(dppe)][PF(6)], 2, have been synthesized and their X-ray crystal structures determined. A related vinylidene complex, [Re(bpy)(CO)(3)(C[triple bond]C[bond]C(6)H(4)[bond](H)C[double bond]C)Fe(C(5)Me(5))(dppe)][PF(6)], 3, has also been synthesized and characterized. The cyclic voltammogram of 1 shows a quasireversible reduction couple at -1.49 V (vs SCE), a fully reversible oxidation at -0.19 V, and a quasireversible oxidation at +0.88 V. In accord with the electrochemical results, density-functional theory calculations on the hydrogen-substituted model complex Re(bpy)(CO)(3)(C[triple bond]C[bond]C(6)H(4)[bond]C[triple bond]C)Fe(C(5)H(5))(dHpe) (Cp = C(5)H(5), dHpe = H(2)P[bond](CH(2))(2)[bond]PH(2)) (1-H) show that the LUMO is mainly bipyridine ligand pi* in character while the HOMO is largely iron(II) d orbital in character. The electronic absorption spectrum of 1 shows low-energy absorption at 390 nm with a 420 nm shoulder in CH(2)Cl(2), while that of 2 exhibits less intense low-energy bands at 432 and 474 nm and additional low-energy bands in the NIR at ca. 830, 1389, and 1773 nm. Unlike the related luminescent rhenium(I)-alkynyl complex [Re(bpy)(CO)(3)(C[triple bond]C[bond]C(6)H(4)[bond]C[triple bond]C[bond]H)], 4, complex 1 is found to be nonemissive, and such a phenomenon is attributed to an intramolecular quenching of the emissive d pi(Re) --> pi*(bpy) (3)MLCT state by the low-lying MLCT and LF excited states of the iron moiety. Interestingly, switching on of the luminescence property derived from the d pi(Re) --> pi*(bpy) (3)MLCT state can be demonstrated in the oxidized species 2 and the related vinylidene analogue 3 due to the absence of the quenching pathway.     

Synthesis of tetracyclic and pentacyclic phenothiazines via benzotriazole methodology

Treatment of 10-(benzotriazol-1-yl)alkylphenothiazine with electron-rich alkenes in the presence of zinc bromide gave tetracyclic phenothiazines 4 and 9 as well as pentacyclic phenothiazines 5 , which have potential use in the pharmaceutical industry.     

Entangled coherent states for quantum information processing

Entangled coherent states (ECSs) with relative phase equal to the phase shift between two coherent states are constructed. We study the degree of entanglement and the nonclassical features exhibited by the so-constructed states keeping in view their role in quantum information processing (QIP).     

Magnetic resonance imaging of cortical bone with ultrashort TE pulse sequences

PurposeNormal adult cortical bone has a very short T₂ and characteristically produces no signal with pulse sequence echo times (TEs) routinely used in clinical practice. We wished to determine whether it was possible to use ultrashort TE (UTE) pulse sequences to detect signal from cortical bone in human subjects and use this signal to characterise this tissue.Subjects and MethodsSeven volunteers and 10 patients were examined using ultrashort TE pulse sequences (TE=0.07 or 0.08 ms). Short and long inversion as well as fat suppression pulses were used as preparation pulses. Later echo images were also obtained as well as difference images produced by subtracting a later echo image from a first echo image. Saturation pulses were used for T₁ measurement and sequences with progressively increasing TEs for T₂* measurement. Intravenous gadodiamide was administered to four subjects.ResultsSignal in cortical bone was detected with UTE sequences in children, normal adults and patients. This signal was usually made more obvious by subtracting a later echo image from the first provided that the signal-to-noise ratio was sufficiently high.Normal mean adult T₁s ranged from 140 to 260 ms, and mean T₂*s ranged from 0.42 to 0.50 ms. T₁ increased significantly with age (P<.01).Increased signal was observed after contrast enhancement in the normal volunteer and the three patients to whom it was administered.Reduction in signal from short T₂ components was seen in acute fractures, and increase in signal in these components was seen with new bone formation after fracture malunion. In a case of osteoporosis, bone cross-sectional area and signal level appeared reduced.ConclusionSignal can be detected from normal and abnormal cortical bone with UTE pulse sequences, and this can be used to measure its T₁ and T₂* as well as observe contrast enhancement. Difference images are of value in increasing the conspicuity of cortical bone and observing abnormalities in disease.     

Tetra[2,3-thienylene]: a building block for single-molecule electromechanical actuators

Tetra(2,3-thienylene) is a thiophene-fused [8]annulene capable of undergoing redox-induced dimensional changes. The feasibility of efficiently translating this intrinsic function into a single polymeric electromechanical actuator is investigated by both experiment and density functional theory (B3LYP 6-31G[d,p]). A study of tetra(2,3-thienylene) and its homodimer reveal that redox-induced conformational change is conserved upon dimerization, a result that implicates similar behavior in the corresponding polymer. Theoretical predictions yield a maximum redox-induced dimensional change of 5.92% per repeat unit for the homodimer. Cyclic voltammetry reveals complete reversibility for the corresponding redox cycle. The latter two facts establish tetra(2,3-thienylene) as a suitable building block for single-molecule electromechanical actuators.     

Functional 2-benzyl-1,2-dihydro[60]fullerenes as acceptors for organic photovoltaics: facile synthesis and high photovoltaic performances

A new series of functional 2-benzyl-1,2-dihydro[60]fullerenes, BnHCs, were synthesized efficiently via co-catalyzed selective monofunctionalization of C₆₀ with functional benzyl bromides. Photophysical and electrochemical properties of the new BnHCs were investigated. PSCs based on 2-MeO–4-CO₂Me–BnHC as new acceptor and P3HT as donor showed a power conversion efficiency of 3.75%, which is comparable to that of PC₆₁BM under the same device conditions.     

Benzimidazole Annulated New Heterocyclic Compounds: Synthesis of Polycyclic Pyrrole Derivatives

Several benzimidazolo[1,2‐a]pyrrole derivative s were obtained by reaction of benzimidazolo[1,2‐a]indeno[2,1‐d]pyrrole derivative with different reactants such as p‐toluenesulfonyl chloride, phenylthiocyanate, phenyl‐isothiocyanate, dimethylsulfate, and active methylene such as malononitrile, ethyl cyanoacetate, benzoyl acetonitrile, ethyl acetoacetate, and diethyl malonate. The newly synthesized products have been deduced on the basis of spectral and analytical data.