Addition of Cl2C: to (-)-O-menthyl acrylate under sonication-phase-transfer catalysis. Efficient synthesis of (+)- and (-)-(2-chlorocyclopropyl)methanol

[reaction: see text] Dichlorocyclopropanation of (-)-O-menthyl acrylate under conditions of phase-transfer catalysis (CHCl3, KOH, tetramethylammonium bromide), with sonication, gives excellent yields (85-94%) of the corresponding dichlorocyclopropanecarboxylate ester compared to thermal conditions (90 degrees C, 56%). No diastereoselectivity was observed, but one isomer was isolated pure by fractional crystallization. The measured kinetic isotope effect (initial rate (CHCl3)/rate (CDCl3) approximately 1.7) suggests deprotonation of CHCl3 as the rate-limiting step.     

Investigation of solvent effects in capillary electrophoresis for the separation of biological porphyrin methyl esters

The effects of organic solvents on the capillary electrophoresis (CE) separation of a number of important biological porphyrin methyl esters - six weakly basic, hydrophobic cyclic tetrapyrroles possessing two and four to eight methyl ester groups around the periphery of the porphyrin ring - were investigated in the mode of micellar electrokinetic chromatography (MEKC), microemulsion electrokinetic chromatography (MEEKC), and nonaqueous CE. In aqueous MEKC, partial separation of the six neutral porphyrin methyl esters was obtained with an organic modifier (acetonitrile) in the concentration range between 20 and 40%, in which sodium dodecyl sulfate (SDS) molecules might be present in the form of SDS micelles and/or SDS micelle-like aggregates. Relatively stable SDS micelles can be formed in nonaqueous MEKC using formamide as the separation medium, but the separation of the target analytes remained unsatisfactory. Improved resolution of all six porphyrin methyl esters was obtained using MEEKC with the running buffer consisting of 0.8% w/w n-heptane (oil phase), 2.25% w/w SDS and 1.0% w/w Brij 35 (mixed surfactant), 6.6% w/w 1-butanol (cosurfactant), and 30% v/v 2-propanol (second cosurfactant), but reproducibility in terms of peak areas for certain porphyrins (especially uroporphyrin I octamethyl ester) was found to be very poor. Best separation performances were achieved with nonaqueous CE separations in which the weakly basic porphyrin methyl esters were protonated under strongly acidic conditions (e.g., using 10 mM perchloric acid) in mixed organic solvents. For example, using a 50:50 mixture of methanol and acetonitrile as the separation medium, baseline separation of all six (positively charged) porphyrin methyl esters can be obtained within 3 min and the average precision (RSD, N = 13) in terms of migration time and peak area were 0.55 and 2.16%, respectively.     

Harderoporphyrin: a misnomer

More than 30 years ago it was reported that rodent Harderian glands contained a tricarboxylic acid porphyrin, which the authors named Harderoporphyrin. The recent finding in rat Harderian glands of the porphyrin glycoconjugate, protoporphyrin-1-O-acyl-beta-xyloside as a major component led to scrutiny of earlier publications. It became apparent that the results were flawed and that the conclusions were unsustainable. The procedural artefacts which led to the errors are discussed and their bases are demonstrated experimentally. Harderoporphyrin as originally defined never existed.     

Tamoxifen metabolism in rat liver microsomes: identification of a dimeric metabolite derived from free radical intermediates by liquid chromatography/mass spectrometry

Tamoxifen has been shown to be a potent liver carcinogen in rats, and generates covalent DNA adducts. On-line high performance liquid chromatography/electrospray ionisation mass spectrometry (HPLC/ESI-MS) has been used to further study the metabolites of tamoxifen formed by rat liver microsomes in the presence of NADPH with a view to identifying potential reactive metabolites which may be responsible for the formation of DNA adducts, and liver carcinogenesis. A metabolite has been detected with a protonated molecule at m/z 773. The mass of this compound is consistent with a dimer of hydroxylated tamoxifen (m/z 388). Analysis of 4-hydroxytamoxifen incubated with a rat liver microsomal preparation showed the formation of a similar metabolite with an apparent MH+ ion at m/z 773, believed to be a dimer of 4-hydroxytamoxifen formed by a free radical reaction. The retention time for this metabolite from 4-hydroxytamoxifen is identical to that of the tamoxifen metabolite, suggesting that these two compounds are the same. The levels of the dimer were higher when 4-hydroxytamoxifen was used as substrate and, in addition, two isomers were detected. It is proposed that tamoxifen was first converted to arene oxides which react with DNA or to 4-hydroxytamoxifen, either directly or via 3,4-epoxytamoxifen, which then undergoes activation via a free radical reaction to give reactive intermediates which can then react with DNA and protein, or with themselves, to give the dimers (m/z 773).     

Analysis of the interaction between alpha-1-acid glycoprotein and tamoxifen and its metabolites

Tamoxifen is administered for the treatment of breast cancer; however resistance to therapy is commonplace. Postulated mechanisms of resistance to tamoxifen include altered pharmacology of the drug, changes in the structure and function of the oestrogen receptor and expression of genes that function to support the growth of cells resistant to tamoxifen. However, binding of drugs to proteins found in the plasma is known to affect the efficacy of drugs and alter their distribution. It is already known that tamoxifen is bound 99% to albumin. We investigated the interaction between the plasma protein, alpha-1-acid glycoprotein (AGP), and tamoxifen, since if binding did occur then the free plasma concentration of the drug would be reduced, resulting in the minimum effective concentration of tamoxifen not being attained. Using a recently described intrinsic fluorescence technique for the study of drug-protein interactions, the extent of binding between tamoxifen citrate and AGP was determined. Furthermore, analysis of binding of the known active metabolites of tamoxifen (4-hydroxytamoxifen, N-desmethyltamoxifen, N-desdimethyltamoxifen, cis-alpha-hydroxytamoxifen and trans-alpha-hydroxytamoxifen) to AGP was conducted. Tamoxifen citrate and metabolites were shown to bind AGP, however the level of interaction was low and negligible at the concentration of the drug found in the plasma.     

Quantitation of midazolam in human plasma by automated chip-based infusion nanoelectrospray tandem mass spectrometry

An automated chip-based infusion nanoelectrospray ionization (nanoESI) platform was used to demonstrate reproducible quantitation of drug molecules from biological matrices. Three sample preparation strategies were explored including protein precipitation of plasma with acetonitrile, de-salting of the plasma, and a combination of protein precipitation with subsequent de-salting of the dried and reconstituted extract. The best results were obtained when fortified human plasma samples containing midazolam were precipitated with acetonitrile containing alprazolam as the internal standard (IS). The supernatant was concentrated to dryness, reconstituted in aqueous acid, and de-salted by automated reversed-phase solid-phase extraction (SPE) prior to infusion nanoESI-MS/MS. Analyses employed a triple quadrupole mass spectrometer operated in selected reaction monitoring (SRM) mode. Each sample was infused for approximately 10 s and the resulting ion current profiles were integrated. Area ratios were used for regression analysis of standard samples (1.5-500 ng/mL). Quality control samples (3, 250, and 400 ng/mL) in five replicates from three different analysis days demonstrated intra-assay precision (< or =16%), inter-assay precision (< or =5%), and overall accuracy (+/-9% deviation). Infusion reproducibility of the assay was established by analyzing extracts after storage for 24 h at ambient temperature. Control plasma samples from six different sources probed the potential utility of this technique for the analysis of clinical samples. At the lower limit of quantitation (LLQ), variability and mean overall accuracy were < or =13% CV and +/-3% deviation, respectively, while at the upper limit of quantitation (ULQ) variability and mean overall accuracy were < or =9% CV and +/-9% deviation, respectively. Inter-chip variability was established by determining standard sample extracts across five different chips (< or =12% CV). Throughput for the assay was 55 s per sample, although this time may be shortened to 40 s per sample with recent improvements in the automated nanoESI system. No contamination or carryover was observed using this promising automated nanoESI-MS/MS platform.     

Synthesis,photophysical and electrochemical properties,and biological labeling studies of cyclometalated iridium(III) bis(pyridylbenzaldehyde) complexes: novel luminescent cross-linkers for biomolecules

We report the synthesis, characterization, photophysical, and electrochemical properties of a series of luminescent cyclometalated iridium(III) complexes containing two aldehyde functional groups [Ir(pba)(2)(N-N)](PF(6)) (Hpba=4-(2-pyridyl)benzaldehyde; N-N=2,2'-bipyridine, bpy (1), 1,10-phenanthroline, phen (2), 3,4,7,8-tetramethyl-1,10-phenanthroline, 3,4,7,8-Me(4)-phen (3), 4,7-diphenyl-1,10-phenanthroline, 4,7-Ph(2)-phen (4)). The X-ray crystal structure of complex 1 has been investigated. Upon photoexcitation, complexes 1-4 exhibit intense and long-lived emission in fluid solutions at 298 K and in low-temperature glass. The luminescence is assigned to a triplet intra-ligand ((3)IL) excited state associated with the pba(-) ligand, probably with mixing of some triplet metal-to-ligand charge-transfer ((3)MLCT) (dpi(Ir)-->pi*(pba(-))) character. Since each of these complexes possesses two aldehyde groups, which can react with the primary amine groups of biomolecules to form stable secondary amines after reductive amination, we have investigated the possibility of these complexes as novel luminescent cross-linkers for biological substrates. L-Alanine has been labeled with complexes 1-4 to give the luminescent bioconjugates 1-(Ala)(2)-4-(Ala)(2). These conjugates show strong photoluminescence with long emission lifetimes under ambient conditions. On the basis of the emission energy trend, the excited state of these luminescent bioconjugates is likely to bear a high parentage of (3)MLCT (dpi(Ir)-->pi*(N-N)) character. In addition, the glycoprotein avidin (Av) has also been conjugated with complexes 1-4 to give the bioconjugates 1-Av-4-Av. Upon photoexcitation, these bioconjugates also display intense and long-lived (3)MLCT (dpi(Ir)-->pi*(N-N)) emission in aqueous buffer at 298 K. Furthermore, a heterogeneous competitive assay for biotin has been developed using 2-Av and biotinylated microspheres. We have shown that complexes 1-4 represent a new class of multicolor luminescent cross-linkers for biomolecular species.     

Carbonate and oxalate dianions as prolific hydrogen-bond acceptors in supramolecular assembly

In the new inclusion compound [(C2H5)4N+]2 x CO3(2-) x 7(NH2)2CS, the carbonate ion is surrounded by twelve convergent NH donor groups from six thiourea molecules to form a hydrogen-bonded aggregate shaped like two concave three-leaved propellers sharing a common core, whereas in [(n-C3H7)4N+]2 x C2O4(2-) x 4(NH2)2CS the oxalate ion serves as a hub for binding four thiourea molecules to generate a cross-shaped structural motif.     

Removal of copper from aqueous solution by aminated and protonated mesoporous aluminas: kinetics and equilibrium

A novel adsorbent, aminated and protonated mesoporous alumina, was prepared and employed for the removal of copper from aqueous solution at concentrations between 5 and 30 mg/l, in batch equilibrium experiments, in order to determine its adsorption properties. The removal of copper by the adsorbents increases with increasing adsorbent dosages. The adsorption mechanism is assumed to be an ion exchange between copper and the hydrogen ions present on the surface of the mesoporous alumina. The adsorbent was characterized by XRD, TEM, SEM, and BET methods. The sorption data have been analyzed and fitted to linearized adsorption isotherm of the Freundlich, Langmuir, and Redlich-Peterson models. The batch sorption kinetics have been tested for first-order, pseudo-first-order, and pseudo-second-order kinetic reaction models. The rate constants of adsorption for all these kinetic models have been calculated. Results also showed that the intraparticle diffusion of Cu(II) on the mesoporous catalyst was the main rate-limiting step.     

An unexpected oxidation of unactivated methylene C-H using DIB/TBHP protocol

An in situ generated hypervalent iodine species, bis(tert-butylperoxy)iodobenzene, was used as a peroxy radical source for the oxidation of unreactive, remote, and isolated alkyl (cyclic or aliphatic) esters and amides to the corresponding keto compounds under very mild conditions.