Abstract: There is increasing in vitro and in vivo evidence indicating that acyl glucuronides of various drugs are chemically reactive and potentially cause organ toxicity. Such conjugates are chemically unstable, undergoing hydrolysis and pH-dependent intramolecular migration to generate isomers and covalent binding with various tissue proteins to result in drug-protein adducts. This review highlights the reactivity of drug acyl glucuronides and commonly used analytical techniques for these metabolites and resultant drug-protein adducts used in preclinical and clinical studies. The stability of acyl glucuronides is dependent on many factors including pH, temperature, nature of the aglycon, and the presence of plasma or albumin. Drug acyl glucuronides may cause toxicity either through changes in the functional properties of the modified proteins, through initiation of antigen-mediated immune responses, or unknown mechanisms. The conjugates and sometimes the drug-protein adducts can achieve appreciable blood concentrations following drug administration. With careful sample handling procedures (e.g. quick cooling and acidification) during preclinical and clinical studies, drug acyl glucuronides in biological matrixes can be reliably identified and quantitated using appropriate analytical methods such as HPLC, LC-MS and NMR techniques.

Abstract: F2-isoprostanes (F 2-isoPs) represent a new family of biomarkers for oxidative stress generated by free radical attack of membrane-bounded arachidonic acid. Esterified F2-isoPs can be found in tissue or plasma lipids whereas the free form F 2-isoPs, hydrolyzed by phospholipase , is mainly present in body fluids. The extent of systematic damage due to oxidative stress within the body can be assessed by the determination of plasma or urine F2-isoPs. The determination of F2-isoPs in clinical practice is not often used due to the complexity to extract the compounds from their biologic matrixes before the analysis step. In most of published protocols, extraction procedure is critical and time- consuming, requiring successive chromatographic steps. Moreover, some of these procedures lead to a substantial loss of target compounds. In order to improve sample preparation steps and final recovery, others methods have been developed and optimized. For detection of F 2-isoPs, two main analytical approaches have been adopted. The first one involves immunological methods and the second approach is based on chromatograph ic separation and detection by mass spectrometry. A large amount of works has been done in the field of isoprostane analysis, but until now, no standardized method seems to emerge. Indeed, described methodologies differ either in the sample preparation steps or in the detection techniques or both. In the present review, the most commonly used methods are presented and compared in terms of extraction, purification, and analysis of F2-isoPs, taking into account the various origins of biological samples.

Abstract: Proteoglycans are ubiquitous biomolecules in the body located in the extracellular matrix, on the cell surface and also within the cells. They contain at least one glycosaminoglycan (GAG) chain covalently attached to a core protein and may also present N- or O-linked glycans. The high structural diversity and distribution relate to the various biological functions of proteoglycans. In recent years, new members have enlarged the proteoglycan family and advances in molecular biology and glycobiology contributed to elucidate more of the biological functions of proteoglycans. In order to study the structure of a proteoglycan molecule and relate it to its function (or dysfunction), its isolation and purification from cell culture or tissue extracts is necessary. Next to the widely used anion exchange chromatographic methods, techniques based on lectin affinity chromatography have created new possibilities to increase the degree of purity. Introduction of electrospray ionization (ESI) and matrix-assisted laser desorption-ionization (MALDI) sources, together with tandem mass spectrometry (MS/MS or MSn), mark a further important step towards the structural analysis of glycosaminoglycans. The aim of this review is to present the most recent advances in proteoglycan purification and analysis.

Abstract: The concept, molecular self-assembly, especially considering molecule-molecule interaction as an information- processing phenomenon, have a profoundly novel effect on thoughts and efforts related to Medicine and Pharmacology. This new style of thinking is still too novice to be used solely and independently for explanation of disease mechanisms and appropriate treatment strategies. However it calls for a range of new researches based on new predictions about disease mechanisms (especially Autoimmune diseases, Endocrinopathies, and Neoplasms) and relevant treatment strategies (superstructural drugs).

Abstract: This review surveys recent publications on the Electrospray Ionisation - Mass Spectrometry (ESI-MS) of drugs of small molecular mass taken from selected drug classes using the Web of Knowledge database. The structural classes are antibiotics/antibacterials, steroids, cannabinols, antidiabetics, immunosuppressants, antitumour drugs, antiretroviral drugs, nonsteroidal antiinflammatories, mucolytic drugs, anticoagulants, cyclooxygenase inhibitors, stimulants, noradrenergic agents, anti-TB drugs and erectile dysfunction agents. Fragmentation information that these drugs exhibit in-source and in ion-trap, triple quadrupole and time-of flight mass spectrometers is also provided. An appraisal of applications for the period 2004-2005, again taken from the Web of Knowledge database, of the technique liquid chromatography - electrospray ionization - mass spectrometry (LC-ESI-MS)to the detection and determination of these drugs, primarily in biomatrices, is then made. Information on such aspects as sample concentration, LC separation conditions, recoveries from biological media and limits of detection (LODs) are provided.