Stable derivatives for the gas chromatographic determination of synthetic anabolic stilbene residues (diethylstilbestrol, dienestrol and hexestrol) in meat and organs of treated cattle in the sub-parts per billion (10(9)) level

A method for the determination of diethylstilbestrol and the related compounds dienestrol and hexestrol residues in meat and organs of treated cattle is described. After extraction and clean-up, these synthetic estrogens are subjected to reaction with pentafluorobenzoyl chloride, which gives very stable perfluoro esters that are suitable for gas chromatographic determination using an electron-capture detector. With the careful clean-up and the very sensitive response of these derivatives, it is possible to reach a limit of detection in the sub-parts per billion (10(9)) range starting with only 5 g of sample.     

Thermal- and pressure-induced cooperative spin transition in the 2D and 3D coordination polymers {Fe(5-Br-pmd)z[M(CN)x]y} (M=AgI, AuI, NiII, PdII, PtII)

A new family of cyanide-based spin-crossover polymers with the general formula {Fe(5-Br-pmd)z[M(CN)x]y} [M=AgI (1), AuI (2), NiII (3), PdII (4), PtII (5); 5-Br-pmd=5-bromopyrimidine; z=1 or 2, x=2 or 4, and y=2 or 1] have been synthesized and characterized using single-crystal X-ray diffraction (XRD), X-ray powder diffraction (XRPD), magnetic susceptibility measurements, and differential scanning calorimetry (DSC). At 293 K, compound 1 presents the monoclinic space group C2/c, whereas at 120 K, it changes to the monoclinic space group P21/c. At 293 K, the crystal structure of 1 displays an uninodal three-dimensional network whose nodes, constituted of FeII, lie at the inversion center of an elongated octahedron. The equatorial bond lengths are defined by the N atoms of four [AgI(CN)2]- groups belonging to two crystallographically nonequivalent AgI atoms, Ag(1) and Ag(2). They are shorter than those of the axial positions occupied by the N atoms of the 5-Br-pmd ligands. The Fe-N average bond length of 2.1657(7) A is consistent with a high-spin (HS) state for the FeII ions. At 120 K, the crystal structure changes refer mainly to the FeII environment. There are two crystallographically independent FeII ions at this temperature, Fe(1) and Fe(2), which adopt the HS and low-spin (LS) states, respectively. The average Fe-N bond length for Fe(1) [2.174(5) A] and Fe(2) [1.955(5) A] agrees well with the reported magnetic data at this temperature. The spin transition of the FeII ions labeled as Fe(1) is found to be centered at Tc downward arrow=149 K and Tc upward arrow=167 K and accompanied by a drastic change of color from orange (HS) to red (LS). Magnetic susceptibility measurements under applied hydrostatic pressure performed on 1 have shown a linear displacement of the transition to higher temperatures while the hysteresis width remains unaltered in the interval of pressures of 105 Pa to 0.34 GPa. A further increase of the pressure induces the spin transition in the Fe(2) ions, which is completely accomplished at 1.12 GPa (T1/2=162 K). Compounds 1 and 2 are isostructural, but 2 does not exhibit spin-transition properties; the FeII centers remain in the HS state in the temperature range investigated, 5-300 K. Compounds 3-5 are not similar or isostructural with 1. A two-dimensional structure for 3-5 has been proposed on the basis of analytical data and the XRPD patterns. Compounds 3-5 undergo first-order spin transition where the critical temperatures for the cooling (Tc downward arrow) and warming (Tc upward arrow) modes are 170 and 180 K (3), 204 and 214 K (4), and 197 and 223 K (5), respectively. It is worth mentioning the color change from yellow to orange observed in 3-5 upon spin transition. The thermodynamic parameters associated with the spin transition estimated from DSC measurements are DeltaH=6 kJ mol(-1) (1), 11 kJ mol(-1) (3), 16 kJ mol(-1) (4), and 16 kJ mol(-1) (5) and DeltaS=38 J K(-1) mol(-1) (1), 62 J K(-1) mol(-1) (3), 76 J K-1 mol(-1) (4), and 81 J K(-1) mol(-1) (5).     

New findings for in-gel digestion accelerated by high-intensity focused ultrasound for protein identification by matrix-assisted laser desorption ionization time-of-flight mass spectrometry

New findings in sample treatment based on high-intensity focused ultrasound (HIFU) for protein digestion after polyacrylamide gel electrophoresis separation are presented. The following variables were studied: (i) sample volume; (ii) sonotrode diameter; (iii) previous protein denaturation; (iv) cooling; (v) enzyme concentration; and (vi) protein concentration. Results showed that positive protein identification could be done after protein separation by gel electrophoresis through peptide mass fingerprint (PMF) in a volume as low as 25 microL. The time needed was less than 2 min and no cooling was necessary. The importance of the sonotrode diameter was negligible. On the other hand, protein denaturation before sonication was a trade-off for the success of procedure here described. The protein coverage was raised from 5 to 30%, and the number of peptides matching the proteins was also increased in a percentage ranging 10-100% when the classical overnight treatment is compared with the proposed HIFU procedure. The minimum amount of protein that can be identified using the HIFU sample treatment by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was 0.06 microg. The lower concentration of trypsin successfully used to obtain an adequate protein digestion was 3.6 microg/mL.     

Influence of Phage Proteins on Formation of Specific UV DMA Photoproducts in Phage T7

Phage T7 can be used as a biological UV dosimeter. Its reading is proportional to the inactivation rate expressed in HT7 units. To understand the influence of phage proteins on the formation of DNA UV photoproducts, cyclobutane pyrimidine dimers (CPD) and (6-4)photoproducts ((6-4)PD) were determined in T7 DNA exposed to UV radiation under different conditions: intraphage T7 DNA, isolated T7 DNA and heated phage. To investigate the effects of various wavelengths, seven different UV sources have been used. The CPD and (6-4)PD were determined by lesion-specific antibodies in an immunodot-blot assay. Both photoproducts were HT7 dose-dependently produced in all three objects by every irradiation source in the biologically relevant UV dose range (1-10 HT7). The CPD to (6-4)PD ratios increased with the increasing effective wavelength of the irradiation source and were similar in intraphage T7 DNA, isolated DNA and heated phage with all irradiation sources. However, a significant decrease in the yield of both photoproducts was detected in isolated T7 DNA and in heated phage compared to intraphage DNA, the decrease was dependent on the irradiation source. Both photoproducts were affected the same way in isolated T7 DNA and heated phage, respectively. The yield of CPD and (6-4)PD was similar in B, C-like and A conformational states of isolated T7 DNA, indicating that the conformational switch in the DNA is not the decisive factor in photoproduct formation. The most likely explanation for modulation of photoproduct frequency in intraphage T7 DNA is that the presence of bound phage proteins induces an alteration in DNA structure that can result in an increased rate of dimerization and (6-4)PD production of adjacent based in intraphage T7 DNA.     

Preparation of Organometallic Ruthenium–Arene–Diaminotriazine Complexes as Binding Agents to DNA

The reactions of two diaminotriazine ligands 2,4‐diamino‐6‐(2‐pyridyl)‐1,3,5‐triazine (2‐pydaT) and 6‐phenyl‐2,4‐diamino‐1,3,5‐triazine (PhdaT) with ruthenium–arene precursors led to a new family of ruthenium(II) compounds that were spectroscopically characterized. Four of the complexes were cationic, with the general formula [(η⁶‐arene)Ru(κ²‐N,N‐2‐pydaT)Cl]X (X=BF₄, TsO; arene=p‐cymene: 1.BF4 , 1.TsO arene=benzene: 2.BF4 , 2.TsO ). The neutral cyclometalated complex [(η⁶‐p‐cymene)Ru(κ²‐C,N‐PhdaT*)Cl] ( 3 ) was also isolated. The structures of complexes 2.BF4 and 3.H2O were determined by X‐ray diffraction. Complex 1.BF4 underwent a partial reversible‐aquation process in water. UV/Vis and NMR spectroscopic measurements showed that the reaction was hindered by the addition of NaCl and was pH‐controlled in acidic solution. At pH 7.0 (sodium cacodylate) Ru–Cl complex 1.BF4 was the only species present in solution, even at low ionic strength. However, in alkaline medium (KOH), complex 1.BF4 underwent basic hydrolysis to afford a Ru–OH complex ( 5 ). Fluorimetric studies revealed that the interaction of complex 1.BF4 with DNA was not straightforward; instead, its main features were closely linked to ionic strength and to the [DNA]/complex ratio. The bifunctional complex 1.BF4 was capable of interacting concurrently through both its p‐cymene and 2‐pydaT groups. Cytotoxicity and genotoxicity studies showed that, contrary to the expected behavior, the complex species was biologically inactive; the formation of a Ru–OH complex could be responsible for such behavior.     

Laser photolysis of 2-phenylheptamethyltrisilane

Two transient absorptions have been detected in the 266 nm laser photolysis of 2-phenylheptamethyltrisilane in cyclohexane solution at room temperature, and their time evolution was recorded in the presence and absence of air and added trapping agents. The shorter-lived 440 nm transient (t1/2 < 20μs) is tentatively assigned to the silylene :SiMePh and the more persistent 380 nm transient to the disilene MePhSiSiMePh. The reactivity of this silylene is much lower than had been expected.     

Intuitionistic nonstandard bounded modified realisability and functional interpretation

We present a bounded modified realisability and a bounded functional interpretation of intuitionistic nonstandard arithmetic with nonstandard principles.The functional interpretation is the intuitionistic counterpart of Ferreira and Gaspar's functional interpretation and has similarities with Van den Berg, Briseid and Safarik's functional interpretation but replacing finiteness by majorisability.We give a threefold contribution: constructive content and proof-theoretical properties of nonstandard arithmetic; filling a gap in the literature; being in line with nonstandard methods to analyse compactness arguments.     

The phenylcarbene rearrangement revisited

The evolution of mechanistic ideas about the phenylcarbene rearrangement has been reviewed, and three closely linked problems have been identified toward whose solution this research has been aimed: 1. Why do the ratios of the stable end products from the rearrangements of o-, m- and p-tolylmethylene differ when all three reactions have been throught to pass through a common intermediate? 2. Why does the rearrangement of 2-methylcycloheptatrienylidene lead to exclusive formation of styrene? 3. What is the mechanism of styrene formation from o-tolylmethylene? New mechanisms have been proposed in which m- and p-tolylmethylene can rearrange to styrene without necessarily being converted to o-tolylmethylene. The formation of a small amount of 2,6-dimethylstyrene from the rearrangement of 3,4,5-trimethylphenylmethylene is viewed as evidence for such a mechanism, and a set of interconverting norcaradienylidenes are believed to be the crucial intermediates. Other alternatives are considered and rejected on the basis of the rearrangement products of 3,5-dimethyl- and 3,4,5-trimethylphenylmethylene.     

Intermediate range order in concentrated aqueous solutions of copper nitrate. X-ray diffraction and Raman investigations

Concentrated aqueous solutions of copper nitrate of different concentrations (0.66 mol dm⁻³ up to 4.84 mol dm⁻³) have been investigated by X-ray diffraction at room temperature. In these solutions a maximum of intensity (prepeak) is observed at low values of Q (approx. 0.6 to 1 Å⁻¹) suggesting the existence of an intermediate range order in their structure, in agreement with previous investigations on different electrolytes. In order to get information on the solvation shell of the copper cation, Raman spectroscopy experiments have been performed for the more concentrated solution. Polarization observations, isotopic substitution of the solvent and the comparison with a Raman spectrum of a concentrated aqueous solution of aluminum nitrate were performed in order to improve the interpretation of the obtained results with these convergent observations.     

X-Ray diffraction investigations of concentrated aqueous solutions of calcium halides

Concentrated aqueous solutions of calcium chloride and calcium bromide have been investigated by X-ray diffraction. In the diffraction patterns of aqueous chloride solutions of different concentrations, a maximum of intensity is observed in the region 0.6–1.1 Å⁻¹, while in the investigated aqueous bromide solution no maximum is observed in that region. This pre-peak suggests the existence of positional correlations in solution beyond direct contact. The interpretation is similar to the one proposed by the authors in previous investigations, and is supported by the calculations based on ad hoc molecular models. The influence of the anion is discussed.