Smart coumarin-tagged imprinted polymers for the rapid detection of tamoxifen.

A signalling molecularly imprinted polymer was synthesised for easy detection of tamoxifen and its metabolites. 6-Vinylcoumarin-4-carboxylic acid (VCC) was synthesised from 4-bromophenol to give a fluorescent monomer, designed to switch off upon binding of tamoxifen. Clomiphene, a chlorinated analogue, was used as the template for the imprinting, and its ability to quench the coumarin fluorescence when used in a 1:1 ratio was demonstrated. Tamoxifen and 4-hydroxytamoxifen were also shown to quench coumarin fluorescence. Imprinted and non-imprinted polymers were synthesised using VCC, methacrylic acid as a backbone monomer and ethylene glycol dimethacrylate as cross-linker, and were ground and sieved to particle sizes ranging between 45 and 25 µm. Rebinding experiments demonstrate that the imprinted polymer shows very strong affinity for both clomiphene and tamoxifen, while the non-imprinted polymer shows negligible rebinding. The fluorescence of the imprinted polymer is quenched by clomiphene, tamoxifen and 4-hydroxytamoxifen. The switch off in fluorescence of the imprinted polymer under these conditions could also be detected under a UV lamp with the naked eye, making this matrix suitable for applications when coupled with a sample preparation system.

N-Aryl N'-hydroxyguanidines, a new class of NO-donors after selective oxidation by nitric oxide synthases: structure-activity relationship.

The formation of nitric oxide (NO) was followed during the oxidation of 37 N-hydroxyguanidines or related derivatives, including 18 new N-aryl N'-hydroxyguanidines, by recombinant inducible nitric oxide synthase (NOS II). Several N-aryl N'-hydroxyguanidines bearing a relatively small, electron-donating para subtituent, such as H, F, Cl, CH(3), OH, OCH(3), and NH(2), led to NO formation rates between 8 and 41% of that of NO formation from the natural NOS substrate, N(omega)-hydroxy-L-arginine (NOHA). The characteristics of these reactions were very similar to those previously reported for the oxidation of NOHA by NOS:(i) the strict requirement of NOS containing (6R)-5,6,7,8-tetrahydro-L-biopterin, reduced nicotinamide adenine dinucleotide phosphate, and O(2) for the oxidation to occur, (ii) the formation of NO and the corresponding urea in a 1:1 molar ratio, and (iii) a strong inhibitory effect of the classical NOS inhibitors such as N(omega)-nitro-L-arginine and S-ethyl-iso-thiourea. Structure-activity relationship studies showed that two structural factors are crucial for NO formation from compounds containing a C(triple bond)NOH function. The first one is the presence of a monosubstituted N-hydroxyguanidine function, since disubstituted N-hydroxyguanidines, amidoximes, ketoximes, and aldoximes failed to produce NO. The second one is the presence of a N-phenyl ring bearing a relatively small, not electron-withdrawing para substituent that could favorably interact with a hydrophobic cavity close to the NOS catalytic site. The k(cat) value for NOS II-catalyzed oxidation of N-para-fluorophenyl N'-hydroxyguanidine was 80% of that found for NOHA, and its k(cat)/K(m) value was only 9-fold lower than that of NOHA. Interestingly, the K(m) value found for NOS II-catalyzed oxidation of N-(3-thienyl) N'-hydroxyguanidine was 25 microM, almost identical to that of NOHA. Recombinant NOS I and NOS III also oxidize several N-aryl N'-hydroxyguanidines with the formation of NO, with a clearly different substrate specificity. The best substrates of the studied series for NOS I and NOS III were N-(para-hydroxyphenyl) and N-(meta-aminophenyl) N'-hydroxyguanidine, respectively. Among the studied compounds, the para-chlorophenyl and para-methylphenyl derivatives were selective substrates of NOS II. These results open the way toward a new class of selective NO donors after in situ oxidation by each NOS family.

Preparation of alpha, beta-acetylenic ketones by catalytic heterogeneous oxidation of alkynes.

Covalent grafting of iron phthalocyanines onto silica affords active catalysts for selective oxidation of alkynes and propargylic alcohols to alpha, beta-acetylenic ketones, highly valuable precursors in the preparation of fine chemicals.

Mannose derived hexacoordinated phosphate--a generally efficient chiral anion for asymmetric applications.

Mannose derived hexacoordinated phosphate--prepared in two steps from methyl-alpha-D-mannopyranoside--is a chiral anionic auxiliary with broad asymmetric efficiency.

Molecularly imprinted polymer solid-phase extraction for detection of zearalenone in cereal sample extracts.

The aim of this work was to develop a method for the clean-up and preconcentration of zearalenone from corn and wheat samples employing molecularly imprinted polymer (MIP) as selective sorbent for solid-phase extraction (SPE). Cereal samples were extracted with acetonitrile/water (75:25, v/v) and the extract was diluted with water and applied to an AFFINIMIP ZON MIP-SPE column. The column was then washed to eliminate the interferences and zearalenone was eluted with methanol and quantified using HPLC with fluorescence detection (lambda(exc)=275/lambda(em)=450 nm). The precision and accuracy of the method were satisfactory for both cereals at the different fortification levels tested and it gave recoveries between 82 and 87% (RSDr 2.5-6.2%, n=3) and 86 and 90% (RSDr 0.9-6.8%, n=3) for wheat and maize, respectively. MIP-SPE column capacity was determined to be not less than 6.6 microg of zearalenone and to be at least four times higher than that of immunoaffinity column (IAC). The application of AFFINIMIP ZON molecularly imprinted polymer as a selective sorbent material for detection of zearalenone fulfilled the method performance criteria required by the Commission Regulation (EC) No. 401/2006, demonstrating the suitability of the technique for the control of zearalenone in cereal samples.

Solid-phase extraction using molecularly imprinted polymers for selective extraction of a mycotoxin in cereals.

The aim of this work was to develop a method for the clean-up of a mycotoxin, i.e. Ochratoxin A (OTA), from cereal extracts employing a new molecularly imprinted polymer (MIP) as selective sorbent for solid-phase extraction (SPE) and to compare with an immunoaffinity column. A first series of experiments was carried out in pure solvents to estimate the potential of the imprinted sorbent in terms of selectivity studying the retention of OTA on the MIP and on a non-imprinted polymer using conventional crushed monolith. The selectivity of the MIP was also checked by its application to wheat extracts. Then, after this feasibility study, two different formats of MIP: crushed monolith and micro-beads were evaluated and compared. Therefore an optimization procedure was applied to the selective extraction from wheat using the MIP beads. The whole procedure was validated by applying it to wheat extract spiked by OTA at different concentration levels and then to a certified contaminated wheat sample. Recoveries close to 100% were obtained. The high selectivity brought by the MIP was compared to the selectivity by an immunoaffinity cartridge for the clean-up of the same wheat sample. The study of capacity of both showed a significant higher capacity of the MIP.

Sugar derived hexacoordinated phosphates: chiral anionic auxiliaries with general asymmetric efficiency.

Mannose-derived hexacoordinated phosphate anions, prepared in as few as two steps from methyl-alpha-D-mannopyranoside and tris(dimethylamino)phosphine, are chiral anionic auxiliaries with broad asymmetric efficiency. These chemically robust anions are effective NMR chiral solvating agents and efficient asymmetry-inducers, able to control the configuration of conformationally labile C2-symmetric monomethinium cations (organic, helical, charge 1+, P or M enantiomers) and D3-symmetric iron(II) trisdiimine complexes (metallo-organic, octahedral, charge 2+, Delta or Lambda enantiomers). Diastereomeric control with often unmatched selective levels was achieved with the help of mannopyranoside backbone of the anions and the substituents on the [1,3]dioxane ring that play a key role in the recognition process, something not obvious at first sight.

Biphasic enantioselective olefin epoxidation using Tropos dibenzoazepinium catalysts.

Several novel chiral iminium TRISPHAT [tris(tetrachlorobenzenediolato)phosphate(V)] salts combining a diphenylazepinium core, chiral exocyclic appendages, and lipophilic counterions have been prepared and tested in biphasic enantioselective olefin epoxidation conditions. Interestingly, the iminium salts derived from commercially available (S)- or (R)-1,2,2-trimethylpropylamine can display efficiency similar to those made from L-acetonamine. Variable-temperature NMR spectroscopy (VT-NMR) and circular dichroism (CD) experiments were performed in search of a correlation between good enantioselectivity in the products and high diastereomeric control of the biphenyl axial chirality of the catalysts.

Inhibitory effects and spectral interactions of isomeric methoxyindazoles on recombinant nitric oxide synthases.

A series of isomeric methoxyindazoles has been evaluated as inhibitors of purified recombinant neuronal, inducible, and endothelial nitric oxide synthases (NOS). 7-Methoxyindazole (7-MI) was the most active compound of this series and displayed selectivity toward the constitutive neuronal (NOS I) and endothelial (NOS III) NOS isoforms, the inducible NOS II being almost insensitive to this inhibitor. 6-, 5-, and 4-Methoxyindazoles were almost inactive against all three NOS isoforms. Inhibition of NO and citrulline formation catalyzed by neuronal NOS in the presence of 7-MI appeared to be competitive versus both substrate L-arginine (L-arg) and (6R)-5,6,7,8-tetrahydrobiopterin (BH(4)) cofactor. 7-MI only slightly inhibited NADPH oxidase activity and was inactive against the cytochrome c (cyt c) reductase activity of neuronal NOS at concentrations up to 100-fold higher than its IC(50) value for inhibition of citrulline formation. UV/Vis and EPR studies indicated that 7-MI interacts with the oxygenase domain of neuronal NOS (NOS I(oxy)) in an identical manner but with a much lower affinity than 7-nitroindazole (7-NI). These results demonstrate that an indazole derivative bearing an electron-rich substituent in the 7-position is also a NOS I inhibitor and that such a compound presents strong similarities with the mechanism of inhibition of 7-NI.