Polycyclic arene sulfides. Preparation of 1a,2,3,11b-tetrahydrobenz-[5,6]anthra[1,2-b]thiirene, 6b,7a,8,9-tetrahydrobenzo[10,11]-chryseno[1,2-b]thiirene,and 7,8,8a,9a-tetrahydrobenzo-[10,11]chryseno[3,4-b]thiirene

The title compounds 5, 7 and 9 which are the first arene sulfides of 7,8-dihydrobenz[a]anthracene, 8,9-and 10,11-dihydrobenzo[a]pyrene, respectively, have been synthesized by treatment of the corresponding arene oxides 4, 6 and 8 with N,N-dimethylthioformamide in the presence of catalytic amounts of trifluoroacetic acid.     

Development of single-chain variable fragment (scFv) antibodies against hapten benzo[a]pyrene: a binding study

Due to its highly carcinogenic and mutagenic effect on humans, a maximum tolerable limit of 10 ng/L of benzo[a]pyrene (B[a]P) in drinking water was set by the European Commission (Council Directive 98/83/EC). Although several polyclonal and monoclonal antibodies (mAb) for the detection of B[a]P and other polycyclic aromatic hydrocarbons (PAH) have been developed by others, a traditional enzyme-linked immunosorbent assay (ELISA) with a limit of quantification of 10 ng/L for monitoring B[a]P has not been developed. With this in mind, several single-chain variable fragment (scFv) antibodies were created using existing mAbs against the extremely hydrophobic hapten B[a]P, and their heavy and light chains recombined to make unique variable light (V_L) and heavy (V_H) chain combinations. Their binding behaviour was investigated using microtiter plate ELISA and surface plasmon resonance techniques. Specifically, the coding sequences for V_L and V_H chains of 10 murine anti-B[a]P antibody producing hybridoma cell lines were isolated by degenerate oligonucleotide primer sets, cloned in phagemid pIT2 and transferred into Escherichia coli HB2151. To systematically investigate the interaction of the V_L and V_H domains, three high-affinity B[a]P-specific and one nonspecific clone were selected and recombined to build a set of 16 different V_L and V_H combinations. On the basis of our data, it was shown that the V_H plays the major role for specific binding of B[a]P, whilst the V_L can, in some cases, increase the final sensitivity of the assay by one order of magnitude. Furthermore, the sequence analysis of scFvs indicates that the complementarity determining region H3 plays a major role in affinity, whilst cross-reactivity to seven other PAHs demonstrates the importance of the V_L in providing cross-reactivity.     

Detection of Benzo[a]pyrene in Fried Food by Ultrasound-Assisted Matrix Solid-Phase Dispersion and Isotope Dilution GC–MS

A simple, sensitive and reliable analytical method was developed for the detection of benzo[a]pyrene in fried food using gas chromatography-mass spectrometry with an isotope-labelled internal standard. Samples were directly extracted and purified by the ultrasound-assisted matrix solid-phase dispersion (MSPD) procedure. The simple pretreatment procedures were tested with different absorbents (C18, NH₂, Oasis, and SiO₂). The optimal ultrasonication-assisted MSPD was achieved by MSPD-SiO₂ and sonication for 10 min in an ultrasonic bath. The samples were quantified using benzo[a]pyrene-d₁₂ as the internal standard. An analysis of the samples spiked with different levels of benzo[a]pyrene showed recoveries ranging from 84.6 to 103.2 %, with an RSD of 3.21–8.32 %, depending on the spiking level. This method was thus shown to be suitable for the detection of benzo[a]pyrene in fried food.     

Fluorescence study of the solubilization of benzo[a]pyrene: application to its detection in coal washing waters

The solubilization of benzo[a]pyrene by organic solvents (dioxane, toluene and dichloromethane) and a surfactant (Triton X-100) was investigated. These media were successfully used for the determination of benzo[a]pyrene using fluorescence detection, with excellent limits of detection and large linear analytical ranges. Benzo[a]pyrene was detected in coal washing waters using liquid chromatography with fluorescence detection. The stability of this compound in dioxane was also examined.     

The characterization of (+/-)-anti-benzo[a]pyrene diolepoxide-DNA adducts and (+/-)-anti-dibenzo[a,l]pyrene diolepoxide--DNA adducts in the same DNA sample using solid-matrix phosphorescence

Solid-matrix phosphorescence (SMP) spectra and lifetimes were used to characterize the (±)-anti-benzo[a]pyrene diolepoxide [(±)-anti-B[a]PDE] and (±)-anti-dibenzo[a,l]pyrene diolepoxide [(±)-anti-DB[a,l]PDE] bonded to the same sample of DNA. SMP spectra and lifetimes were also acquired for two samples of DNA that had only (±)-anti-B[a]PDE or (±)-anti-DB[a,l]PDE bonded to the individual samples of DNA. A detailed comparison of the SMP properties was made among the three samples of DNA. The SMP excitation spectra for the (±)-anti-B[a]PDE-DNA and the (±)-anti-DB[a,l]PDE-DNA adducts were very similar. However, the SMP emission spectra of the two DNA adduct systems were very dissimilar with a major emission band for the (±)-anti-B[a]PDE-DNA adducts appearing at 613 nm and for the (±)-anti-DB[a,l]PDE-DNA adducts a major band was at 558 nm. It was possible to selectively use SMP emission wavelengths and obtain a SMP excitation of spectrum of the (±)-anti-DB[a,l]PDE-DNA adducts in the dual adducted DNA sample without the (±)-anti-B[a]PDE-DNA adducts emitting SMP. In addition, it was shown that the SMP emission spectrum of the dual adducted DNA sample could be used to detect both adduct systems in the modified DNA sample. It was demonstrated that the SMP lifetimes could be effectively employed to characterize the dual adducted DNA sample. For example, the SMP decay curve for the (±)-anti-DB[a,l]PDE-DNA adducts could be acquired without any SMP emission from the (±)-anti-B[a]PDE-DNA adducts. Also, ln(SMP intensity) versus time plots were very useful in characterizing the dual adducted DNA sample.     

Room-temperature excitation-emission phosphorescence matrices and second-order multivariate calibration for the simultaneous determination of pyrene and benzo[a]pyrene

The present article describes the simultaneous phosphorimetric determination of pyrene and benzo[a]pyrene, two highly toxic polycyclic aromatic hydrocarbons, through excitation-emission phosphorescence matrices (EEPMs) and second-order calibration. The developed approach enabled us to determine both compounds at μg L⁻¹ concentration levels without the necessity of applying separation steps, as well as significantly reducing the experimental time. An artificial neural network (ANN) approach was applied to optimize the chemical variables which have an influence on the room-temperature phosphorescence emission of the studied analytes. The present study was employed for the discussion of the scopes of the applied second-order chemometric tools: parallel factor analysis (PARAFAC) and partial least-squares with residual bilinearization (PLS/RBL). The superior capability of PLS/RBL to model the profiles of other potentially interferent polycyclic aromatic hydrocarbons (PAHs) was demonstrated. The quality of the proposed method was established with the determination of both pyrene and benzo[a]pyrene in artificial and real water samples.     

Thermodynamic study of (anthracene + benzo[a]pyrene) solid mixtures

To characterize better the thermodynamic behavior of a binary polycyclic aromatic hydrocarbon mixture, thermochemical and vapor pressure experiments were used to examine the phase behavior of the {anthracene (1) + benzo[a]pyrene (2)} system. A solid–liquid phase diagram was mapped for the mixture. A eutectic point occurs at x₁ = 0.26. The eutectic mixture is an amorphous solid that lacks organized crystal structure and melts between T = (414 and 420) K. For mixtures that contain 0.10 < x₁ < 0.90, the enthalpy of fusion is dominated by that of the eutectic. (Solid + vapor) equilibrium studies show that mixtures of anthracene and benzo[a]pyrene at x₁ < 0.10 sublime at the vapor pressure of pure benzo[a]pyrene. These results suggest that the (solid + vapor) equilibrium of benzo[a]pyrene is not significantly influenced by moderate levels of anthracene in the crystal structure.     

Tight Molecular Recognition of Benzo[a]pyrene by a High‐Affinity Antibody

Benzo[a ]pyrene, which is produced during the incomplete combustion of organic material, is an abundant noxious pollutant because of its carcinogenic metabolic degradation products. The high‐affinity (K _D≈3 nm ) monoclonal antibody 22F12 allows facile bioanalytical quantification of benzo[a ]pyrene even in complex matrices. We report the functional and X‐ray crystallographic analysis of 22F12 in complex with 3‐hydroxybenzo[a ]pyrene after cloning of the V‐genes and production as a recombinant Fab fragment. The polycyclic aromatic hydrocarbon is bound in a deep pocket between the light and heavy chains, surrounded mainly by aromatic and aliphatic amino acid side chains. Interestingly, the hapten–antibody interface is less densely packed than expected and reveals polar, H‐bond‐like interactions with the polycyclic aromatic π‐electron system, which may allow the antibody to maintain a large, predominantly hydrophobic binding site in an aqueous environment while providing sufficient complementarity to its ligand.     

A novel application of nylon membranes to the luminescent determination of benzo[a]pyrene at ultra trace levels in water samples

Very simple and highly sensitive methods are presented for the determination of benzo[a]pyrene, one of the most carcinogenic polycyclic aromatic hydrocarbons (PAHs). The approaches are based on solid-phase extraction of the analyte on a nylon membrane via a syringe procedure, and its fluorescent or phosphorescent determination on the solid surface. While the native fluorescence of benzo[a]pyrene retained on a nylon surface is measured directly, room-temperature phosphorescence is induced by spotting a few microlitres of thallium(I) nitrate solution on the surface (heavy-atom effect). An enhancement of the phosphorescence signal was corroborated when the measurements were carried under a nitrogen atmosphere. The analytical figures of merit obtained under the best experimental conditions demonstrate the capability of detecting benzo[a]pyrene at a sub-parts-per-trillion (sub-ng L⁻¹) level. The potential interference from other common PAHs and also from different metal ions was studied. The feasibility of determining benzo[a]pyrene in real samples was successfully evaluated through the analysis of spiked tap, underground and mineral water samples of different origins. Recoveries obtained from spiked river waters were successfully compared with those provided by a reference method, through rigorous statistical analysis.     

The electrogenerated chemiluminescence of pyrene and some related compounds

The characteristics of the electrogenerated chemiluminescence in dimethylformamide of pyrene, 1,2-benzpyrene, 3,4-benzpyrene and dibenzo[a,lpyrene, dibenzo[a,i]pyrene, dibenzo [a,h]pyrene and dibenzo [b,l]pyrene generated with optimum conditions of applied potential are described. The appearance of emission at wavelengths longer than those expected from the excited singlet state of the hydrocarbons may be attributed to decomposition products. Pyrene and 3,4-benzpyrene exhibit long wavelength emission, however, which it is suggested may be due to excimer formation.     

Molecular structures,electronic spectra,and aromaticity of 1,6- and 1,8-dioxa analogues of pyrene and benzo[a]pyrene. Semiempirical molecular orbital calculations

The molecular structures of [1]benzopyrano[6,5,4-def][1]benzopyran, [2]benzopyrano[7,8,1,def]-1-benzopyran, 1-benzopyrano[6,5,4-mna]xanthene, and 2-benzopyrano[7,8,1-mna]xanthene have been optimized by using the PM3 semiemperical MO method. These calculated molecules have been shown to be planar. Their aromaticity has been investigated by use of the HOMA index. The molecules are found to be less aromatic than the correspolding parednt hydrocarbons, pyrene and benzo[a]pyrene. The CNDO/S MO method has been used to interpret th experimental uv-vis specrtoscopic data. The result of the PM3, CNDO/S and HOMA-index clculations are in good arteement with the experimental data.     

Estimation of the Sampling Error in the Determination of Benzo[a]pyrene in Organized Gas–Dust Air Emissions

The components and the total error of the determination of benzo[a]pyrene in gas–dust flows from organized sources of various technological processes of heat-and-power, aluminum, and building-material production; petroleum chemistry; and heating systems using low-temperature luminescence were studied. The dependences of the sampling errors on the factors of their formation (flow temperature; rate, time, and volume of the sampled material; the type of sorbent; and filtration method) were determined. The conditions favorable to the improvement of the sampling precision were found.     

Synthesis of 5,6-Dimethylbenz[a]phenazine

5,6-Dimethylbenz[a]phenazine ( 4 ), an aza analogue of the carcinogenic 5,6-dimethylbenz-[c]acridine has been obtained by a 1,1-dehydration-rearrangement (Wagner-Meerwin type) from 5,5-dimethyl-6-hydroxy-5,6-dihydrobenz[a]phenazine ( 3 ). The alcohol 3 was obtained from the hydrogenation of the corresponding ketone 2 which was prepared in two ways: Method A, the oxidation of 5,5-dimethyl-5,6-dihydrobenz[a]phenazine ( 1 ); Method B, the hydrolysis of the 6,6-dibromo derivative 5 of 1.     

Light induced interactions of benzo[a]pyrene with carboxylic acids

Abstract The in vitro photochemical behaviour of benzo[a]pyrene (BP) in presence of short and long chain carboxylic acids is studied. The direct irradiation (295–400 or 320–400 nm) of BP in solution in the presence of saturated carboxylic acids destroys more than 70% of this carcinogen in 72 h with a 150 W Xe arc lamp. Consumption of BP alone under similar conditions is less than 10%. The principal product resulting from direct interaction of BP with carboxylic acids is characterized as 6-acyloxybenzo[a]pyrene by means of UV, IR, NMR and mass spectroscopy. The other pathways of destruction may involve polymerization. Saturated fatty acids are shown to favour photoacyloxyation, while unsaturated fatty acids appear to facilitate polymerization of BP. The effect of different factors, such as the carboxylic acid involved, solvent, and wavelength on photoacyloxylation reaction as well as on the consumption of BP is discussed. Photoinduced interaction of BP with carboxylic acids seems to involve radical cation as well as free radicals of the hydrocarbon. Biological properties of BP esters in terms of primary irritancy and carcinogenic activity have also been tested by mouse skin assay. Both long and short chain esters do not show any carcinogenic activity; conversely, long chain esters are shown to be more severe irritants as compared to short chain esters or BP.     

Synthesis of pyrazolo[1,5‐a]‐, 1,2,4‐triazolo[1,5‐a]‐ and imidazo[1,2‐a]pyrimidines related to zaleplon,a new drug for the treatment of insomnia

This paper describes the preparation of some pyrazolo[1,5‐a]‐, 1,2,4‐triazolo[1,5‐a]‐ and imidazo[1,2‐a]‐pyrimidines substituted on the pyrimidine moiety by a 4‐[(N‐acetyl‐N‐ethyl)amino]phenyl group. A new synthesis of related benzo[h]pyrazolo[1,5‐a]‐, benzo[h]pyrazolo[5,1‐b]‐ and benzo[h]1,2,4‐triazolo[1,5‐a]‐quinazolines is also reported.     

Synthesis of derivatives of the new heterocyclic systems benz[f] imidazo [2,1 -a ] isoquinoline and benz [f] pyrimido [2,1 -a] isoquinoline

The synthesis of several derivatives of benz [f] imidazo [2,1 -a] isoquinoline and benz [f] pyrimido-[2,l-a]isoquinoline, both new heterocyclic structures, is described.     

The synthesis of 7,8-dihydroimidazo[1,2-e]pyrazolo[1,5-a]-1,3,5-triazines, 8,9-dihydro-7H-pyrimido[1,2-e]pyrazolo[1,5-a]-1,3,5-triazines and 7,8,9,10-tetrahydro[1,3]diazepino[1,2-e]pyrazolo[1,5-a]triazines

Cyclic hydrazino amidines were converted to the corresponding aminopyrazolyl derivatives. Ring closure between the amino groups of pyrazoline moieties and NH groups of cyclic amidines afforded the following ring systems: 7,8-Dihydroimidazo[1,2-e]pyrazolo[1,5-a]-1,3,5-triazines, 8,9-dihydro-7H-pyrimido[1,2-e]pyrazolo[1,5-a]-1,3,5-triazines and 7,8,9,10-tetrahydro[1,3]diazepino[1,2-e]pyrazolo[1,5-a]-1,3,5-triazines.     

On triazoles XXIV . Synthesis of cycloalka[1,2,4]triazolo[1,5-a]pyrimidinones

The uv and nmr spectral data of some 6,7,8,9,10,11-hexahydrocyclohepta[d][1,2,4]triazolo[1,5-a]pyrimidin-5-one, 5,6,7,8,9,11-hexahydrocyclohepta[e][1,2,4]triazolo[1,5-a]pyrimidin-10-one, 6,7,8,9,10,11-hexahydrocycloocta[d][1,2,4]triazolo[1,5-a]pyrimidin-5(12H)-one, 5,6,7,8,9,10-hexahydrocycloocta[e][1,2,4]triazolo[1,5-a]-pyrimidin-11(12H)-one, 6,7,8,9,10,11,12,13,14,15-decahydrocyclododeca[d][1,2,4]triazolo[1,5-a]pyrimidin-5(16H)-one and 5,6,7,8,9,10,11,12,13,14-decahydrocyclododeca[e][1,2,4]triazolo[1,5-a]pyrimidin-15(16H)-one as well as their N-benzylated derivatives representing six novel ring systems were compared to prove their structure. The N-benzylation of the highly insoluble cyclic amides to yield the isomeric N-benzyl derivatives 3/1, 3/2 and 3/3 distinguished by INAPT was performed through their readily soluble tetrabutylammonium salts.     

The synthesis of [1]benzothieno[2,3-c]quinolines, [1]benzothieno[2,3-c][1,2,4]triazolo[4,3-a]quinoline,and [1]benzothieno[2,3-c]tetrazolo[1,5-a]quinoline

Photocyclization of 3-chloro-N-phenylbenzo[b]thiophene-2-carboxamide 10 afforded [1]benzothieno[2,3-c]-quinolin-6(5H)-one 11 which was chlorinated to 6-chloro[1]benzothieno[2,3-c]quinoline 12 followed by dechlorination to give [1]benzothieno[2,3-c]quinoline 5 . A series of 6-substituted alkoxy and thioalkoxy[1]benzothieno[2,3-c]quinoline derivatives were prepared along with the N-methyl quaternary salt 13 of 5 . 6-Chloro[1]-benzothieno[2,3-c]quinoline 12 was converted into 6-hydrazino[1]benzothieno[2,3-c]quinoline 23 which upon treatment with formic acid yielded [1]benzothieno[2,3-c][1,2,4]triazolo[4,3-a]quinoline 6 . Treatment of 23 with nitrous acid resulted in [1]benzothieno[2,3-c]tetrazolo[1,5-a]quinoline 7 . Compounds 6 and 7 are novel heterocyclic ring systems.     

The syntheses of 4b,5a-dihydro-5H-benzo[3,4]-phenanthro[1,2-b]azirine and 1a,11b-dihydro-6,11-dimethyl-1H-benz[3,4]anthra[1,2-b]azirine

The syntheses of the K-imine derivatives of carcinogenic benzo[c]phenanthrene and 7,12-dimethylbenz-[a]anthracene are described. Treatment of trans-5-azido-5,6-dihydrobenzo[c]phenanthren-6-ol ( 3 ) and trans-6-azido-5,6-dihydrobenzo[c]phenanthren-5-ol ( 5 ) with thionyl chloride yielded the corresponding β-chloro azides, which in turn, were reacted with lithium aluminium hydride to give 4b,5a-dihydro-5H-benzo[3,4]-phenanthro[1,2-b]azirine ( 2 ). In a similar manner trans-5-azido-5,6-dihydro-7,12-dimethylbenz[a]anthracen-6-ol ( 11 ) and trans-6-azido-5,6-dihydro-7,12-dimethylbenz[a]anthracen-5-ol ( 13 ) were transformed to the respective chloro azides and, converted into 1a,11b-dihydro-6,11-dimethyl-1H-benz[3,4]anthra[1,2-b]azirine ( 10 ).