Improved design for high resolution electrospray ionization ion mobility spectrometry.

An improved design for high resolution electrospray ionization ion mobility spectrometry (ESI-IMS) was developed by making some salient modifications to the IMS cell and its performance was investigated. To enhance desolvation of electrospray droplets at high sample flow rates in this new design, volume of the desolvation region was decreased by reducing its diameter and the entrance position of the desolvation gas was shifted to the end of the desolvation region (near the ion gate). In addition, the ESI source (both needle and counter electrode) was positioned outside of the heating oven of the IMS. This modification made it possible to use the instrument at higher temperatures, and preventing needle clogging in the electrospray process. The ion mobility spectra of different chemical compounds were obtained. The resolving power and resolution of the instrument were increased by about 15-30% relative to previous design. In this work, the baseline separation of the two adjacent ion peaks of morphine and those of codeine was achieved for the first time with resolutions of 1.5 and 1.3, respectively. These four ion peaks were well separated from each other using carbon dioxide (CO(2)) rather than nitrogen as the drift gas. Finally, the analytical parameters obtained for ethion, metalaxyl, and tributylamine indicated the high performance of the instrument for quantitative analysis.

Alternatives for coupling sequential injection systems to commercial capillary electrophoresis-mass spectrometry equipment.

On-line coupling of an automated flow system with a commercially available capillary electrophoresis (CE) system with an electrospray interface (ESI) for mass spectroscopic (MS) detection is described. The peculiarities of CE-ESI-MS interfaces, in which a high electrical field must be applied to the capillary end where the sample is provided by the flow system, introduce significant difficulties for the appropriate work of the entire arrangement. Experimental strategies are proposed for achieving stable conditions for on-line sample pre-treatment, conditioning of the separation capillary, sample injection, as the proper separation. The versatility and robustness of the proposed arrangement is discussed, taken as example the separation of a variety of amines. Connection of the CE system's pressure to the automated flow system enables hydrodynamic introduction of sample with high precision. The developed hyphenated system is of practical relevance as it opens an avenue for the simplification and automation of the whole analytical process required when using powerful CE-ESI-MS equipments.

High-throughput mass spectrometer using atmospheric pressure ionization and a cylindrical ion trap array.

The analytical performance of an atmospheric pressure sampling, multiple-channel, high-throughput mass spectrometer was investigated using samples of a variety of types. The instrument, based on an array of cylindrical ion traps, was built with four independent channels and here is operated using two fully multiplexed channels (sources, ion optics, ion traps, detectors) capable of analyzing different samples simultaneously. Both channels of the instrument were incorporated within the same vacuum system and operated using a common set of control electronics. A multichannel electrospray ionization source was assembled and used to introduce samples including solutions of organic compounds, peptides, and proteins simultaneously into the instrument in a high-throughput fashion. Cross-talk between the channels of the instrument occurred in the detection system and could be minimized to 1-2% using shielding between detector channels. In this initial implementation of the instrumentation, an upper mass/charge limit of approximately 1300 Th was observed (+13 charge state of myoglobin) and unit mass/charge resolution was achieved to approximately 800 Th. The rather limited dynamic range (2-3 orders of magnitude for low-concentration analytes) is due to cross-talk contributions from more concentrated species introduced into a different channel. Analysis of mixtures of alkylamines and peptides is demonstrated, but analysis of mixtures with a wide spread in mass/charge ratios was not possible due to mass discrimination in the ion optics. Further refinement of the vacuum system and ion optics will allow the addition of more channels of parallel mass analysis and facilitate applications in fields such as proteomics and metabolomics.

Enzymatic asymmetric synthesis of alpha-methyl arylalkylamines and alpha-methyl arylalkylalcohols by arylalkyl acylamidases.

With the novel microbial enzyme, 'arylalkyl acylamidase', optically active alpha-methyl arylalkylamines and alpha-methyl arylalkylalcohols have been obtained through enantioselective hydrolysis of their racemic amides and esters. (S)-Enantiomers of 1-methylbenzylamine, 1-methyl-3-phenylpropylamine and 1-methyl-3-phenylpropanol of high optical purity (> 94% e.e.) were synthesized with the cells of Nocardia erythropolis IAM 1440 or Cellulomonas fimi AKU 671. (R)-Enantiomer of 1-methyl-3-phenylpropylamine and (S)-enantiomer of 1-methyl-2-phenylpropanol of high optical purity (> 95% e.e.) were synthesized with the crude preparation of arylalkyl acylamidase of Pseudomonas putida Sc2 AKU 881.

Pharmacokinetic profile of tert-butylaminoethanethiol

A preliminary study of the pharmacokinetic parameters of t-Butylaminoethanethiol (TBAESH) was performed after administration of a single dose (35 mg/kg) either orally or intravenously. Plasma or blood samples were treated with dithiothreitol, perchloric acid and, after filtration, submitted to further purification with anionic resin. In the final step the drug was retained on a cationic resin column, eluted with NaCl lM and detected according to the method of Ellman (1958). The results suggested a pharmacokinetic behavior related to a one open compartment model with the following values for the total drug: area under the intravenous curve (AUC i.v.): 443(+ ou -) 24.0; AUC oral: 85.5(+ ou -) 14.5 ug min.ml(elevado a -1); elimination rate constant: 0.069(+ ou -) 0.0055 min(elevado a -1), biological half-life: 10.0(+ ou -) 0.80 min; distribution volume 1.15(+ ou -) 0.15 ml/g; biodisponibility: 0.19(+ ou -) 0.02. From a pharmacokinetic standpoint, TBAESH seems to have no advantage over the analogous disulfide compound

Gas chromatographic analysis of bacterial amines as their free bases.

Columns of Chromosorb 103, Tenax-GC, Amine 220 plus potassium hydroxide on Chromosorb W, and Carbowax 20M plus potassium hydroxide on Chromosorb W were compared for their ability to separate bacterial amines as their free bases in aqueous solution. A 1.52 m X 0.6 cm O.D. column of Chromosorb 103 separated eleven amines when operated isothermally at 185 degrees C. A further four high-boiling amines could be separated at 240 degrees C. The other packings separated only eight amines isothermally, except for Tenax-GC which separated seven of the free bases. Chromosorb 103 performed less well than Carbowax 20 M plus potassium hydroxide with respect to number of plates or peak resolution. The maximum number of amines separated, thirteen, required Chromosorb 103 programmed from 170 degrees C to 230 degrees C at 3 degrees C min-1 after an initial holding time of 20 min. It was possible tentatively to identify amines in culture supernatant fluid of Proteus mirabilis, viz. ethylamine, isobutylamine and isoamylamine, after direct injection of culture supernatant fluid.

4-Aminobutyraldehyde dehydrogenase activity in rat brain.

An enzyme with NAD+-dependent 4-aminobutyraldehyde dehydrogenase activity was purified about 360-fold from rat brain extract. AMP-Sepharose chromatography was effective in separating the enzyme from other NAD+-dependent aldehyde dehydrogenases included in the extract. The KmS for the substrates NAD+ and 4-aminobutyraldehyde were 4.8 x 10(-4) and 8.3 x 10(-5) M, respectively. The pH optimum for the enzyme was about 8.0. The ratio of activities toward 4-aminobutyraldehyde, propionaldehyde, succinate semialdehyde, and benzaldehyde was 1.00:0.17:0.24:0.09:0.03 when the activity toward 4-aminobutyraldehyde was set equal to 1.00. The enzyme activity in subcellular fractions of rat brain was localized in cytosol.