The study of highly air-and moisture-sensitiVe organometallic complexes by ESI-MS is greatly facilitated by interfacing the instrument to an inert-atmosphere gloVebox, a modification that requires no special precautions beyond proximity.

This is the first study presenting a multi-residue method allowing for comprehensive analysis of several chiral pharmacologically active compounds (cPACs) including beta-blockers, antidepressants and amphetamines in wastewater and digested sludge at the enantiomeric level. Analysis of both the liquid and solid matrices within wastewater treatment is crucial to being able to carry out mass balance within these systems. The method developed comprises filtration, microwave assisted extraction and solid phase extraction followed by chiral liquid chromatography coupled with tandem mass spectrometry to analyse the enantiomers of 18 compounds within all three matrices. The method was successfully validated for 10 compounds within all three matrices (amphetamine, methamphetamine, MDMA, MDA, venlafaxine, desmethylvenlafaxine, citalopram, metoprolol, propranolol and sotalol), 7 compounds validated for the liquid matrices only (mirtazapine, salbutamol, fluoxetine, desmethylcitalopram, atenolol, ephedrine and pseudoephedrine) and 1 compound (alprenolol) passing the criteria for solid samples only. The method was then applied to wastewater samples; cPACs were found at concentration ranges in liquid matrices of: 1.7 ng L(-1) (metoprolol) - 1321 ng L(-1) (tramadol) in influent,

It is now apparent that each of the known, naturally occurring polyphosphoinositides, the phosphatidylinositol monophosphates (PtdIns3P, PtdIns4P, PtdIns5P), phosphatidylinositol bisphosphates [PtdIns(3,4)P 2 , PtdIns(3,5)P 2 , PtdIns(4,5)P 2 ], and phosphatidylinositol trisphosphate [PtdIns(3,4,5) ]inositol radiolabeling, acidified lipid extraction, deacylation, and ion-exchange head group separation, which are time-consuming and not suitable for samples in which radiolabeling is impractical, thus greatly restricting the study of these lipids in many physiologically relevant systems. Thus, we have developed a novel, high-efficiency, buffered citrate extraction methodology to minimize acid-induced phosphoinositide degradation, together with a high-sensitivity liquid chromatography-mass spectrometry (LC-MS) protocol using an acetonitrile-chloroform-methanol-water-ethylamine gradient with a microbore silica column that enables the identification and quantification of all phosphoinositides in a sample. The liquid chromatograph is sufficient to resolve PtdInsP 3 and PtdInsP 2 regioisomers; however, the PtdInsP regioisomers require a combination of LC and diagnostic fragmentation to MS 3 . Data are presented using this approach for the analysis of phosphoinositides in human platelet and yeast samples. As our understanding of lipids as highly dynamic structures with both structural and signaling roles matures, one group stands out. These are the phosphoinositides, which regulate a host of cellular events, such as membrane trafficking, secretion, adhesion, migration, cell survival, and replication (1-4). Many of these outcomes are regulated by interactions between phosphoinositides and effector proteins bearing specific phosphoinositide binding domains (e.g., ENTH, FYVE, PH, PX) (5). Within this lipid family, the best known are phosphatidylinositol (PtdIns), the parent of the higher phosphorylated forms, which itself has no known signaling role, phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P 2 ], and phosphatidylinositol 3,4,5-trisphosphate [PtdIns(3,4,5)P 3 ]. PtdIns(4,5)P 2 is the precursor to diacylglycerol and inositol 1,4,5-trisphosphate in receptor signaling and also directly regulates cytoskeletal reorganizations and the activities of enzymes such as phospholipase D. PtdIns(3,4,5)P 3 regulates cell movement, apoptosis, metabolism, and proliferation via the activation of phosphoinositide dependent kinase 1, protein kinase B, and other PH and PX domain proteins. However, it is now also apparent that other PtdInsP 2 regioisomers, such as phosphatidylinositol 3,4-bisphosphate [PtdIns(3,4)P 2 ] and phosphatidylinositol 3,5-bisphosphate [PtdIns(3,5)P 2 ], together with the phosphatidylinositol monophosphate regioisomers phosphatidylinositol 3-phosphate (PtdIns3P), phosphatidylinositol 4-phosphate (PtdIns4P), and phosphatidylinositol 5-phosphate (PtdIns5P), have their own specific functions. PtdIns3P and PtdIns4P regulate membrane trafficking, whereas PtdIns5P has been reported to bind to the PHD domain i...

Liquid chromatography combined with electrospray ionization mass spectrometry (LC/ESI-MS) has been used successfully for the characterization of biomolecules in proteomics in the last few years. This methodology relied largely on the use of reversed-phase chromatography, in particular C18-based resins, which are suitable for separation of peptides. Here we show that polymeric [polystyrene divinylbenzene] monolithic columns can be used to separate peptide mixtures faster and at a higher resolution. For 500 fmol bovine serum albumin, up to 68% sequence coverage and Mascot Mowse scores of >2000 were obtained using a 9 min gradient on a monolithic column coupled to an ion trap mass spectrometer with ultra-fast MS/MS scan rates. In order to achieve similar results using C18 columns, it was necessary to extend gradient times to 30 min. In addition, we demonstrate the utility of this approach for the analysis of whole Escherichia coli cell lysates by one-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (1D-SDS-PAGE) in combination with LC/MS/MS using 4 min gradients on monolithic columns. Our results indicate higher throughput capabilities of monolithic columns (3-fold gain in time or more) for conventional proteomics applications, such as protein identification and high sequence coverage usually required for detection of post-translational modifications (PTMs). Further optimization of sensitivity and quality of sequence information is discussed, in particular when combined with mass spectrometers that have very fast MS-MS/MS switching and scanning capabilities.

Abstract. The composition of organic aerosol formed from the gas phase ozonolysis of cyclohexene has been investigated in a smog chamber experiment. Comprehensive gas chromatography with time of flight mass spectrometric detection was used to determine that dicarboxylic acids and corresponding cyclic anhydrides dominated the small gas phase reaction products found in aerosol sampled during the first hour after initial aerosol formation. Structural analysis of larger more polar molecules was performed using liquid chromatography with ion trap tandem mass spectrometry. This indicated that the majority of identified organic mass was in dimer form, built up from combinations of the most abundant small acid molecules, with frequent indication of the inclusion of adipic acid. Trimers and tetramers potentially formed via similar acid combinations were also observed in lower abundances. Tandem mass spectral data indicated dimers with either acid anhydride or ester functionalities as the linkage between monomers. High-resolution mass spectrometry identified the molecular formulae of the most abundant dimer species to be C