Isolation of the retinal isomers from the isomerization of all-trans-retinal by flash countercurrent chromatography

The isolation of the retinal isomers from all-trans-retinal was performed by flash countercurrent chromatography. In each separation, isomerization reaction solution of 200 mg all-trans-retinal could be loaded on a 1200 mL of high-speed countercurrent chromatographic column with 5 mm bore, eluted by a mobile phase flow rate of 25 mL/min, resulting in 63 mg of 11-cis-retinal, 24 mg of 13-cis-retinal and 26 mg of 9-cis-retinal with purities more than 95%. n-Hexane–acetonitrile (3:1) was used as the solvent system which possesses the advantages of simplicity, re-use of the solvent and multiple injections. This method could be used to prepare 13-cis-retinal, 11-cis-retinal and 9-cis-retinal for the photoisomerization investigation, such as the effect of 11-cis-retinal in the visual system.     

Capillary electrophoresis-based assessment of nanobody affinity and purity

Drug purity and affinity are essential attributes during development and production of therapeutic proteins. In this work, capillary electrophoresis (CE) was used to determine both the affinity and composition of the biotechnologically produced “nanobody” EGa1, the binding fragment of a heavy-chain-only antibody. EGa1 is an antagonist of the epidermal growth factor receptor (EGFR), which is overexpressed on the surface of tumor cells. Using a background electrolyte (BGE) of 50 mM sodium phosphate (pH 8.0) in combination with a polybrene-poly(vinylsulfonic acid) capillary coating, CE analysis of EGa1 showed the presence of at least three components. Affinity of the EGa1 components towards the extracellular domain of EGFR was assessed by adding different concentrations (0–12 nM) of the receptor to the BGE while measuring the effective electrophoretic mobility of the respective EGa1 components. Binding curves obtained by plotting electrophoretic mobility shifts as a function of receptor concentration, yielded dissociation constants (K_d) of 1.65, 1.67, and 1.75 nM for the three components, respectively; these values were comparable to the K_d of 2.1 nM obtained for the bulk EGa1 product using a cellular assay. CE with mass spectrometry (MS) detection using a BGE of 25 mM ammonium acetate (pH 8.0) revealed that the EGa1 sample comprised of significant amounts of deamidated, bisdeamidated and N-terminal pyroglutamic acid products. CE–MS using a BGE of 100 mM acetic acid (pH 2.8) in combination with a polybrene–dextran sulfate–polybrene capillary coating demonstrated the additional presence of minor products related to incomplete removal of the signal peptide from the produced nanobody. Combining the results obtained from affinity CE and CE–MS, it is concluded that the EGa1 nanobody product is heterogeneous, comprising highly-related proteins that exhibit very similar affinity towards EGFR.     

Synthesis, characterization and Schlenk equilibrium studies of methylmagnesium compounds with O- and N-donor ligands - the unexpected behavior of [MgMeBr(pmdta)] (pmdta = N,N,N′,N″,N″-pentamethyldiethylenetriamine)

MgMe₂ (1) was found to react with 1,4-diazabicyclo[2.2.2]octane (dabco) in tetrahydrofuran (thf) yielding a binuclear complex [{MgMe₂(thf)}₂(μ-dabco)] (2). Furthermore, from reactions of MgMeBr with diglyme (diethylene glycol dimethyl ether), NEt₃, and tmeda (N,N,N′,N′-tetramethylethylenediamine) in etheral solvents compounds MgMeBr(L), (L = diglyme (5); NEt₃ (6); tmeda (7)) were obtained as highly air- and moisture-sensitive white powders. From a thf solution of 7 crystals of [MgMeBr(thf)(tmeda)] (8) were obtained. Reactions of MgMeBr with pmdta (N,N,N′,N″,N″-pentamethyldiethylenetriamine) in thf resulted in formation of [MgMeBr(pmdta)] (9) in nearly quantitative yield. On the other hand, the same reaction in diethyl ether gave MgMeBr(pmdta) · MgBr₂(pmdta) (10) and [{MgMe₂(pmdta)}₇{MgMeBr(pmdta)}] (11) in 24% and 2% yield, respectively, as well as [MgMe₂(pmdta)] (12) as colorless needle-like crystals in about 26% yield. The synthesized methylmagnesium compounds were characterized by microanalysis and ¹H and ¹³C NMR spectroscopy. The coordination-induced shifts of the ¹H and ¹³C nuclei of the ligands are small; the largest ones were found in the tmeda and pmdta complexes. Single-crystal X-ray diffraction analyses revealed in 2 a tetrahedral environment of the Mg atoms with a bridging dabco ligand and in 8 a trigonal-bipyramidal coordination of the Mg atom. The single-crystal X-ray diffraction analyses of [MgMe₂(pmdta)] (12) and [MgBr₂(pmdta)] (13) showed them to be monomeric with five-coordinate Mg atoms. The square-pyramidal coordination polyhedra are built up of three N and two C atoms in 12 and three N and two Br atoms in 13. The apical positions are occupied by methyl and bromo ligands, respectively. Temperature-dependent ¹H NMR spectroscopic measurements (from 27 to −80 °C) of methylmagnesium bromide complexes MgMeBr(L) (L = thf (4); diglyme (5); NEt₃ (6); tmeda (7)) in thf-d₈ solutions indicated that the deeper the temperature the more the Schlenk equilibria are shifted to the dimethylmagnesium/dibromomagnesium species. Furthermore, at −80 °C the dimethylmagnesium compounds are predominant in the solutions of Grignard compounds 4-6 whereas in the case of the tmeda complex7 the equilibrium constant was roughly estimated to be 0.25. In contrast, [MgMeBr(pmdta)] (9) in thf-d₈ revealed no dismutation into [MgMe₂(pmdta)] (12) and [MgBr₂(pmdta)] (13) even up to −100 °C. In accordance with this unexpected behavior, 1:1 mixtures of 12 and 13 were found to react in thf at room temperature yielding quantitatively the corresponding Grignard compound 9. Moreover, the structures of [MgMeBr(pmdta)] (9c), [MgMe₂(pmdta)] (12c), and [MgBr₂(pmdta)] (13c) were calculated on the DFT level of theory. The calculated structures 12c and 13c are in a good agreement with the experimentally observed structures 12 and 13. The equilibrium constant of the Schlenk equilibrium (2 9c ? 12c + 13c) was calculated to be K_gas = 2.0 × 10⁻³ (298 K) in the gas phase. Considering the solvent effects of both thf and diethyl ether using a polarized continuum model (PCM) the corresponding equilibrium constants were calculated to be K_thf = 1.2 × 10⁻³ and K_ether = 3.2 × 10⁻³ (298 K), respectively.     

Universal fluorescent tri-probe ligation equipped with capillary electrophoresis for targeting SMN1 and SMN2 genes in diagnosis of spinal muscular atrophy

This is the first ligase chain reaction used for diagnosis of spinal muscular atrophy (SMA). Universal fluorescent tri-probe ligation (UFTPL), a novel strategy used for distinguishing the multi-nucleotide alternations at single base, is developed to quantitatively analyze the SMN1/SMN2 genes in diagnosis of SMA. Ligase chain reaction was performed by adding three probes including universal fluorescent probe, connecting probe and recognizing probe to differentiate single nucleotide polymorphisms in UFTPL. Our approach was based on the two UFTPL products of survival motor neuron 1 (SMN1) and SMN2 genes (the difference of 9 mer) and analyzed by capillary electrophoresis (CE). We successfully determined various gene dosages of SMN1 and SMN2 genes in homologous or heterologous subjects. By using the UFTPL-CE method, the SMN1 and SMN2 genes were fully resolved with the resolution of 2.16 ± 0.37 (n = 3). The r values of SMN1 and SMN2 regression curves over a range of 1–4 copies were above 0.9944. Of the 48 DNA samples, the data of gene dosages were corresponding to that analyzed by conformation sensitive CE and denatured high-performance liquid chromatography (DHPLC). This technique was found to be a good methodology for quantification or determination of the relative genes having multi-nucleotide variants at single base.     

Metal complexes stability constant determination by hyphenation of capillary electrophoresis with inductively coupled plasma mass spectrometry: The case of 1:1 metal-to-ligand stoichiometry

Nuclear energy development has raised new issues like radionuclides biogeochemistry. The modelling of their biochemical properties involves the accurate determination of thermodynamical data, like stability constants. This can be achieved by using hyphenated capillary electrophoresis (CE)–ICPMS and the method was applied successfully on 1:1 lanthanum–oxalate and uranyl–oxalate complexes. Several significant steps are discussed: choice of analytical conditions, electrophoretic mobility calculation, mathematical treatment of experimental data by using linear regressions, ligand concentration and ionic strength corrections. The following values were obtained with a good precision for lanthanum–oxalate and uranyl–oxalate complexes: log(K°(LaOxa⁺)) = 6.10 ± 0.10 and log(K°(UO₂Oxa)) = 6.40 ± 0.30, respectively, at infinite dilution. These values are consistent with the literature data, showing CE–ICPMS potential for metal complexes stability constants determination.     

Alkyl and dialkylammonium tetrafluoroborate catalyzed cis-trans isomerization of 1,3,5-trimethyl-1,3,5-triphenylcyclotrisiloxane

Alkyl and dialkylammonium tetrafluoroborate promoted cis-trans isomerization of 1,3,5-trimethyl-1,3,5-triphenylcyclotrisiloxane (1) in DMSO-d₆ were studied. The isomerization equilibrium constant K are within the range of 3.74-3.30 from 22 to 47 °C. Thermodynamic parameters of ΔH° and ΔS° for the isomerization were −0.95 kcal/mol and −0.59 cal/mol-K respectively. The isomerization rate is first order in [cis-1] and second order in [R_nNH_4−nBF₄]. Both components of R_nNH_4−n⁺ and BF₄⁻ are essential for the catalytic cis-trans isomerization. The catalytic strength follows the decreasing order of ⁺H₃N(CH₂)₆NH₃⁺>n-C₈H₁₇NH₃⁺>n-C₁₆H₃₃NH₃⁺>Me₃CNH₃⁺>PhCH₂NH₃⁺>Et₂NH₂⁺?Ph₂CHNH₃⁺, Et₃NH⁺. Inversion region was observed in the plot of ln(k_f/T) versus (1/T) with the ceiling located at around 38 °C. The positive activation enthalpy of 9 kcal/mol was estimated at 22-32 °C. The activation enthalpy turns to be slightly negative at T>38 °C.     

Coordination to copper(II) strongly enhances the in vitro antimicrobial activity of pyridine-derived N(4)-tolyl thiosemicarbazones

Five copper(II) complexes with N(4)-ortho, N(4)-meta and N(4)-para-tolyl thiosemicarbazones derived from 2-formyl and 2-acetylpyridine were obtained and thoroughly characterized. The crystal structure of N(4)-meta-tolyl-2-acetylpyridine thiosemicarbazone (H2Ac4mT) was determined, as well as that of its copper(II) complex [Cu(2Ac4mT)Cl], which contains an anionic ligand and a chloride in the coordination sphere of the metal. The in vitro antimicrobial activities of all thiosemicarbazones and their copper(II) complexes were tested against Salmonella typhimurium and Candida albicans. Upon coordination a substantial decrease in the minimum inhibitory concentration, from 225 to 1478 μmol L⁻¹ for the thiosemicarbazones to 5–30 μmol L⁻¹ for the complexes was observe against the growth of Salmonella typhimurium and from 0.7–26 to 0.3–7 μmol L⁻¹ against the growth of C. albicans, suggesting that complexation to copper(II) could be an interesting strategy of dose reduction.     

Formation of γ-Fe2O3 nanoparticles and vacancy ordering: An in situ X-ray powder diffraction study

The formation of maghemite, γ-Fe₂O₃ nanoparticles has been studied by in situ X-ray powder diffraction. The maghemite was formed by thermal decomposition of an amorphous precursor compound made by reacting lauric acid, CH₃(CH₂)₁₀COOH with Fe(NO₃)₃·9H₂O. It has been shown that cubic γ-Fe₂O₃ was formed directly from the amorphous precursor and that vacancy ordering starts about 45 min later at 305 °C resulting in a tripled unit cell along the c-axis. The kinetics of grain growth was found to obey a power law with growth exponents n equal to 0.136(6) and 0.103(5) at 305 and 340 °C, respectively. Particles with average sizes of 12 and 13 nm were obtained in 86 and 76 min at 305 and 340 °C, respectively. The structure of cubic and vacancy ordered phases of γ-Fe₂O₃ was studied at 305 °C by Rietveld refinements.     

Absorption and emission behaviour of trans-p-coumaric acid in aqueous solutions and some organic solvents

The absorption and fluorescence behaviour of trans-p-coumaric acid (trans-4-hydroxycinnamic acid) is investigated in buffered aqueous solution over a wide range from pH 1 to pH 12, in un-buffered water, and in some organic solvents. Absorption cross-section spectra, fluorescence quantum distributions, fluorescence quantum yields, and degrees of fluorescence polarisation are measured. p-Coumaric acid exists in different ionic forms in aqueous solution depending on the pH. There is an equilibrium between the neutral form (p-CAH₂) and the single anionic form (p-CAH⁻) at low pH (pK_na ≈ 4.9), and between the single anionic and the double anionic form (p-CA²⁻) at high pH (pK_aa ≈ 9.35). In the organic solvents studied trans-p-coumaric acid is dissolved in its neutral form. The fluorescence quantum yield of trans-p-coumaric acid in aqueous solution is ?_F ≈ 1.4 × 10⁻⁴ for the neutral and the single anionic form, while it is ?_F ≈ 1.3 × 10⁻³ for the double anionic form. For trans-p-coumaric acid in organic solvents fluorescence quantum yields in the range from 4.8 × 10⁻⁵ (acetonitrile) to 1.5 × 10⁻⁴ (glycerol) were measured. The fluorescence spectra are 7700–10,000 cm⁻¹ Stokes shifted in aqueous solution, and 5400–8200 cm⁻¹ Stokes shifted in the studied organic solvents. Decay paths responsible for the low fluorescence quantum yields are discussed (photo-isomerisation and internal conversion for p-CA²⁻, solvent-assisted intra-molecular charge-transfer or ππ^∗ to nπ^∗ transfer and internal conversion for p-CAH₂ and p-CAH⁻). The solvent dependence of the first ππ^∗ electronic transition frequency and of the fluorescence Stokes shift of p-CAH₂ is discussed in terms of polar solute–solvent interaction effects. Thereby the ground-state and excite-state molecular dipole moments are extracted.     

K2Mo4O13 phases prepared by hydrothermal synthesis

Hydrothermal synthesis in the K-Mo oxide system was investigated as a function of the pH of the reaction medium. Four compounds were formed, including two K₂Mo₄O₁₃ phases. One is a new low-temperature polymorph, which crystallizes in the orthorhombic, space group Pbca, with Z=8 and unit cell dimensions a=7.544(1) Å, b=15.394(2) Å, c=18.568(3) Å. The other is the known triclinic K₂Mo₄O₁₃, whose structure was re-determined from single crystal data; its cell parameters were determined as a=7.976(2) Å, b=8.345(2) Å, c=10.017(2) Å, α=107.104(3)°, β=102.885(3)°, γ=109.760(3)°, which are the standard settings of the crystal lattice. The orthorhombic phase converts endothermically into triclinic phase at ca. 730 K with a heat of transition of 8.31 kJ/mol.     

Synthesis of dihydropyrano[3,2-e]indoles as rotationally restricted phenolic analogs of 5-hydroxyindole—thermal Claisen approach versus gold catalysis

The Claisen rearrangement/cyclization of 5-propargyloxyindoles (2) to afford dihydropyrano[3,2-e]indoles (3) as direct precursors to tetrahydropyrano[3,2-e]indoles (1, a rotationally restricted phenolic analog of 5-hydroxyindole) was examined using either refluxing bromobenzene (156 °C) or Au⁺¹ catalysis in refluxing dioxane (101 °C). This transformation was best effected using Au⁺¹ catalysis (i.e., tris[triphenylphosphinegold(I)] oxonium tetrafluoroborate) because this method required a lower reaction temperature and gave better yields when compared to the simple thermal reaction conditions (156 °C).     

Temperature dependent selective synthesis of linear 2-bromo and 2-alkoxyfuro[2,3-b]quinolines: reaction of 3-(2,2-dibromovinyl-)quinolin-2(1H)-ones with alcoholic KOH

Base promoted and temperature dependent reactions of 3-(2,2-dibromovinyl)quinolin-2(1H)-ones have been described. At 50 °C, the cyclization reactions afforded 2-bromofuro[2,3-b]quinolines in good yields. However, at elevated temperature (80 °C) the domino reaction proceeded affording 2-alkoxyfuro[2,3-b]quinolines via cyclization and nucleophilic substitution reactions.     

Two-stage formation of chloro (N-o-chlorobenzamido-meso-tetraphenylporphyrinato) zinc(II) methylene chloride solvate: Zn(N-NHCO(o-Cl)C6H4-tpp)Cl · CH2Cl2

The coordinating properties of N-o-chlorobenzamido-meso-tetraphenylporphyrin (N-NHCO(o-Cl)C₆H₄-Htpp; 11) have been investigated for the Zn²⁺ ion. Insertion of Zn results in the formation of the zinc complex Zn(N-NCO(o-Cl)C₆H₄-tpp)(MeOH) · MeOH (12 · MeOH). The diamagnetic 12 · MeOH can be transformed into the diamagnetic Zn(N-NHCO(o-Cl)C₆H₄-tpp)Cl · CH₂Cl₂ (13 · CH₂Cl₂) in a reaction with aqueous hydrogen chloride (2%). X-ray structures for 12 · MeOH and 13 · CH₂Cl₂ have been determined. The coordination sphere around the Zn²⁺ ion in 12 · MeOH is a distorted trigonal bipyramid with N(2), N(4) and O(2) lying in the equatorial plane, whereas for the Zn²⁺ ion in 13 · CH₂Cl₂, it is a square-based pyramid in which the apical site is occupied by the Cl(1) atom.     

Microwave-assisted cycloadditions of 2-alkynylbenzonitriles with sodium azide: selective synthesis of tetrazolo[5,1-a]pyridines and 4,5-disubstituted-2H-1,2,3-triazoles

Under microwave irradiation (75 W), treatment of 2-alkynylbenzonitriles with 1.5 equiv of sodium azide in DMSO at 140 °C gave 4,5-disubstituted-2H-1,2,3-triazoles in 60-99% yields. Additionally, adding 8 equiv of ZnBr₂ and using 8 equiv of sodium azide in DMF at 100 °C lead to the formation of tetrazolo[5,1-a]isoquinolines up to 87% yield.     

Determination of ethylenediaminetetraacetic acid, ethylenediaminedisuccinic acid and iminodisuccinic acid in cosmetic products by capillary electrophoresis and high performance liquid chromatography

A capillary electrophoresis (CE) and a high performance liquid chromatography (HPLC) method are described for the simultaneous determination of ethylenediaminetetraacetic acid (EDTA), S,S′-ethylenediaminedisuccinic acid (EDDS) and R,S-iminodisuccinic acid (IDS) complexing agents as their Fe(III) complexes in cosmetics like shower cream and foam bath. The non-biodegradable EDTA is used in combination with biodegradable analogues like EDDS and IDS in many commercial products. The HPLC method involves separation by reversed-phase ion pair chromatography on a C₁₈ column using methanol-formate buffer (20 mM tetrabutylammonium hydrogen sulfate, 15 mM sodium formate adjusted to pH 4.0 with formic acid) (10:90, v/v) as mobile solvent at a flow rate of 0.8 mL min⁻¹ at 24 °C using UV detection at 240 nm. The CE separation was performed in a fused silica capillary of 50 μm i.d. with the total length of 50 cm with a 10 mM MES and MOPSO (pH 5.5) at an applied voltage of −25 kV. The samples were introduced by applying a 50 mbar pressure for 2 s. Absorbances at 215 and 225 nm were monitored for the detection of the complexes. The methodology performance of the two methods was evaluated in terms of linearity, limit of detection (LOD), limit of quantitation (LOQ) and reproducibility. The LOD values obtained from HPLC are low when compared with CE. The applicability of both the methods was demonstrated for the analysis of cosmetic products such as shower cream and foam bath. The results obtained by both CE and HPLC were found to be comparable and in good agreement.     

Electrostatic forces that determine O,O-trans versus O,S-cis conformers in the aminated porphyrin complexes of Cd(N-NHCO-2-C4H3O-tpp)(OAc) and Cd(N-NHCO-2-C4H3S-tpp)(OAc)

Two new diamagnetic, mononuclear and aminated porphyrin complexes of O,O-trans-Cd (3-trans) and O,S-cis-Cd (4-cis) have been synthesized and characterized by ¹H, ¹³C NMR spectroscopy. The crystal structures of (acetato)(N-2-furancarboxamido-meso-tetraphenylporphyrinato)cadmium(II) [Cd(N-NHCO-2-C₄H₃O-tpp)(OAc); 3-trans] and (acetato)(N-2-thiophenecarboxamido-meso-tetraphenylporphyrinato)cadmium(II) [Cd(N-NHCO-2-C₄H₃S-tpp)(OAc); 4-cis] were determined. The coordination sphere around Cd²⁺ is a distorted square-based pyramid in which the apical site is occupied by a bidentate chelating OAc⁻ group for 3-trans and 4-cis. The plane of three pyrrole nitrogen atoms [i.e., N(1), N(2), N(4) for 3-trans and N(1), N(2), N(3) for 4-cis] strongly bonded to Cd²⁺ is adopted as a reference plane 3N. The N(3) and N(4) pyrrole rings bearing the 2-furancarboxamido (Fr) and 2-thiophenecarboxamido groups in 3-trans and 4-cis, respectively, deviate mostly from the 3N plane, thus orienting separately with a dihedral angle of 33.4° and of 31.0°. In 3-trans, Cd²⁺ and N(5) are located on different sides at 1.06 and −1.49 Å from its 3N plane, while in 4-cis, Cd²⁺ and N(5) are also located on different sides at 1.04 and −1.53 Å from its 3N plane. An attractive electrostatic interaction between the Cd²⁺ and O(4)⁻ atoms in furan stabilizes the O,O-trans conformer of 3. A repulsive electrostatic interaction between Cd²⁺ and S(1)⁺ destabilizes the O,S-trans conformer of 4. Both of these repulsive and the mutually attractive interactions between S(1)⁺ and O(3)⁻ atoms favor the O,S-cis rotamer of 4 both in the vapor phase and in low polarity solvents. NOE difference spectroscopy, HMQC and HMBC were employed for the unambiguous assignment of the ¹H and ¹³C NMR resonances of 3-trans and 4-cis in CDCl₃ at 20 and −50 °C.     

Crystal structure,spectroscopy and magnetism of a dinuclear strongly antiferromagnetically coupled N-oxido-bridged Cu(II) compound with 2,2′-bipyridine-1,1′-dioxide (bpdo) as a ligand; [Cu2(bpdo)2Br4]

A new dinuclear compound, [Cu₂(bpdo)₂Br₄], (in which bpdo = 2,2′-Bipyridine-1,1′-dioxide), has been synthesized and fully characterized, including the X-ray and the magnetic susceptibility. Each copper(II) ion in the dinuclear compound has a distorted square pyramidal geometry with the basal plane formed by two oxygen atoms of two ligand molecules which are bridging between the Cu ions with Cu–O distances of 2.021(2) and 2.039(2) Å and two bromide atoms with Cu–Br distances of 2.3577(6) and 2.3665(7) Å. The fifth position is occupied by a non bridging oxygen atom of a ligand with a Cu–O distance of 2.197(2) Å. The distance between the Cu ions is 3.334 Å, while the Cu–O–Cu angle is 110.37(9)°. The magnetic susceptibility measurements (from 5 to 350 K) agree with a very strong antiferromagnetic interaction with a large singlet–triplet splitting (J) of −905 cm⁻¹. At high T (above 250 K) a triplet powder EPR is observed.     

Reactivity of M(en)Cl2 (M = Pd/Pt,en = 1,2-diaminoethane) with N,N′-bis(salicylidene)-p-phenylenediamine: Binding with hexafluorobenzene

Schiff base N,N′-bis(salicylidene)-p-phenylenediamine (LH₂) complexed with Pt(en)Cl₂ and Pd(en)Cl₂ provided [Pt(en)L]₂ · 4PF₆ (1) and Pd(Salen) (2) (Salen = N,N′-bis(salicylidene)-ethylenediamine), respectively, which were characterized by their elemental analysis, spectroscopic data and X-ray data. A solid complex obtained by the reaction of hexafluorobenzene (hfb) with the representative complex 1 has been isolated and characterized as 3 (1 · hfb) using UV–Vis, NMR (¹H, ¹³C and ¹⁹F) data. A solid complex of hfb with a reported Zn-cyclophane 4 has also been prepared and characterized 5 (4 · hfb) for comparison with complex 3. The association of hfb with 1 and 4 has also been monitored using UV–Vis and luminescence data.     

Quantitative study of stereospecific binding of monoclonal antibody to anti-benzo(a)pyrene diol epoxide-N-dG adducts by capillary electrophoresis immunoassay

The stereospecific binding of monoclonal antibody (mAb) 8E11 to anti-benzo(a)pyrene diol epoxide (BPDE)-dG adducts in single nucleoside, long oligonucleotide, and genomic DNA were quantitatively evaluated using noncompetitive and competitive capillary electrophoresis (CE) immunoassays. Two single-stranded TMR-BPDE-90mers containing a single anti-BPDE-dG adduct with defined stereochemistry and a fluorescent label at 5′-end were used as fluorescent probes for competitive CE immunoassay. To quantitatively evaluate the binding affinity through competitive CE immunoassays, a series of equations were derived according to the binding stoichiometry. The binding of mAb 8E11 to trans-(+)-anti-BPDE-dG displays strongest affinity (K_b: 3.57 × 10⁸ M⁻¹) among all four investigated anti-BPDE-dG mononucleoside adducts, and the cis-(−)-anti-BPDE-dG displays lowest affinity (K_b: 1.14 ×10⁷ M⁻¹). The binding of monoclonal antibody (mAb) 8E11 to BPDE-dG adducts in long DNA (90mer) preferentially forms the complex with a stoichiometry of 1:1, and that mAb 8E11 displays a slightly higher affinity with trans-(+)-anti-BPDE-90mers (K_b: 6.36 ± 0.54 × 10⁸ M⁻¹) than trans-(−)-anti-BPDE-90mers (K_b: 4.52 ± 0.52 × 10⁸ M⁻¹). The mAb 8E11 also displays high affinity with BPDE-dG adducts in genomic DNA (K_b: 3.74 × 10⁸ M⁻¹), indicating its promising applications for sensitive immuno-detection of BPDE-DNA adducts in genomic DNA.     

Gas-phase thermolysis of 1-acylnaphtho[1,8-de][1,2,3]triazines. Interesting direct routes towards condensed naphtho[1,8-de]heterocyclic ring systems

FVP pyrolysis of 1-acylnaphtho[1,8-de][1,2,3]triazines at 500 °C and 10⁻² Torr gave exclusively the corresponding 2-substituted naphtho[1,8-de][1,3]oxazines. The latter was also obtained by static pyrolysis but in lower yield along with the corresponding N-(naphthalen-1-yl)acylamides. The reaction was studied kinetically and mechanistically.