PubMed Health⌕ Search

Biomedical subjects

M E Szulc

Publications and source records attributed to M E Szulc.

4 recordsLinked to original sources

Reduction of pantethine in rabbit ocular lens homogenate.

In several animal models, preliminary studies have indicated that pantethine may inhibit cataract formation. Therefore, preclinical trials need to be conducted to study the pharmacology of pantethine in the ocular lens and to establish its efficacy. Since pantethine, which is a disulfide, can undergo a variety of chemical modifications such as reduction and formation of mixed disulfides, a detailed study was first conducted to determine the stability of pantethine in rabbit lens homogenate. A knowledge of the stability of pantethine in lens homogenate was necessary to establish if pantethine could be metabolized in the time it takes to harvest and homogenize a lens. The results of this study will be used to establish a protocol for harvesting and homogenizing lens samples. Pantethine (100 microM) is completely reduced to pantetheine in rabbit lens homogenate in about 16 min. About 1.5% of the pantethine added to lens homogenate forms a mixed disulfide with lens proteins, and the remainder is found in the supernatant. The supernatant pantethine concentration decreases exponentially as a function of time, and the terminal half-life for this process is 3.3 min. The free supernatant pantetheine concentration increases in pseudo first order manner as a function of time with a rate constant of 4.3 min. Pantethinase activity is not significant, because the free supernatant pantetheine concentration did not decrease. The exact mechanism of pantethine reduction in rabbit lens homogenate remains to be determined.

Animals↗

Solid-phase derivatization reactions for biomedical liquid chromatography.

Polymeric reagents have been developed for performing off- and on-line derivatizations of numerous organic analytes in HPLC-detection modes. Such reagents utilize ionic or covalent attachment of labile tags that possess specific detector enhancement properties: ultraviolet, electrochemical, fluorescence, and so forth. Specific synthetic procedures have evolved to generate various linkages of the tag to the underlying, polymeric support, usually involving activated ester connections (leashes). The polymer itself may play a number of roles in the nature of the overall reactions, such as hydrophobic-hydrophillic exclusion, pore size restriction, stabilization of the attachment leashes, and protection of the tags from hydrolysis in aqueous media. The basic, underlying chemistry of polymeric reagents has evolved to the point where it is possible to engineer the polymer support itself, the attachment leash, and the various tags that are then transferred to the analyte molecules. These procedures have now reached the stage of commercialization and practical applicability for real-world drugs and bioorganics in complex biofluid type samples. Polymer supported reagents can now be used for direct injection of biofluids with solid-phase (hydrophobic) extraction of the analytes of interest, followed by sample cleanup, derivatization, elution onto the HPLC column, peak compression, gradient HPLC elution, multiple detection, and final data interpretation with quantitation. This review summarizes much or most of what has been described in the scientific literature over the past decade in the various areas where polymeric reagents are being used for derivatization in HPLC and in capillary electrophoresis as well.

Chemistry Techniques, Analytical↗

Improved detection and derivatization in capillary electrophoresis.

Capillary electrophoresis is well known for its low mass detectabilities, but suffers from poor concentration detection limits. This review will discuss improvements in concentration detectability with an emphasis on derivatization methods. Sample concentration techniques and improved detector designs will also be discussed. Pre- and post-capillary derivatization methods for biofluid analytes such as amino acids, peptides, proteins, oligonucleotides, and oligosaccharides will be examined in detail.

Amino Acids↗

Mixed-bed polymeric o-nitrobenzophenone reagents for the on-line derivatization of amines in high performance liquid chromatography.

Several polymer-bound o-nitrobenzophenone reagents containing different detector-sensitive tags have been combined in the same reactor for the on-line derivatization of amine samples. The formation of multiple derivatives allows numerous opportunities for quantitation from the same injection, and also allows improved identification from the retention times of the multiple derivatives. Changing the reaction conditions changes the ratio of the products formed, so that changes in the ratio of peak heights and areas can also be used for analyte identification. In this work, propylamine was derivatized in acetonitrile on-line, precolumn. Changing the reaction conditions, of reaction time, temperature, solvent, presence of catalyst and the components of the reactor, changed the ratio of the derivatives formed. These changes in product formation with changing reaction conditions were applied to the identification and quantitation of amphetamine and methamphetamine in urine. The drugs were identified by the retention times of their derivatives, the ratio of the peak areas of the derivatives and the change in the ratios after changing reaction conditions. Each derivative was also used for quantitation of levels of spiked concentrations of amphetamine and methamphetamine, with relative errors less than 8%.

Amines↗