Electrospray ion source with a dynamic liquid flow splitter.

RATIONALE: Although the electrospray method is widely used in biomedical mass spectrometry and ion-drift spectrometry, the electrospray system can generate large drops that are the cause of electrical breakdowns, and of the fast clogging of the inlet diaphragm of the differential pumping first stage and system of ion transportation to the analyzer. Thus, a system free of such problems would be of benefit. METHODS: To drain the condensate away from the electrospraying zone and to maintain stability of spraying, a source was designed where the excessive non-sprayed solution flowing down over the metal capillary outer wall is pumped away by an air micropump. An anti-electrode is mounted opposite the metal capillary and is slightly tilted with respect to the horizon to make the condensed liquid flow down. RESULTS: An electrospray ion source allowing work with chromatographic flows of the solution under normal conditions was tested. The applicability of the operating modes of the electrospray ion source with dynamic liquid flow splitting for both ion-drift and mass spectrometers was demonstrated. The flow rate range of liquid fed to the electrospray ion source with dynamic liquid flow splitting is such that it can be used in high performance liquid chromatography/Ion Mobility Spectrometry (HPLC/IMS) and HPLC/MS in the online mode. CONCLUSIONS: An electrospray ion source with a dynamic liquid flow splitter was developed and tested. Optimal operating modes of the electrospray ion source with dynamic liquid flow splitting were determined.

[Identification of rat and human hemoglobin acetilation sites after its interaction with acetylsalicylic acid].

The possibility of interaction of 0.1 mg/mL acetylsalicylic acid with purified human and rat globin in vitro during 24 h at 37 degrees C was investigated. The rat globin can be modified with acetylsalicylic acid on aminoacid residues K-17, K-57, K-91, K-140 in alpha subunit as well as on K-18, K-77 in beta subunit. The human globin can be modified with acetylsalicylic acid on aminoacid residues K-17, K-41, K-57 and K-91 in alpha subunit as well as on K-18, K-96 and K- 133 in beta subunit. We identified of acetetylated lysines K-17 and K-57 in alpha subunit of human hemoglobin after incubation whole blood with 0.1 mg/mL acetylsalicylic acid during 3 h.

ESI-MS studies of silver ion competitive interaction with cysteine-containing peptides and sulfur-containing amino aсids.

The paper deals with investigation of silver ion interaction with sulfur-bearing amino acids and cysteine-bearing peptides using an electrospray ionisation orthogonal ion introduction time-of-flight (ESI-o-TOF) mass spectrometer. It has been shown that Cys and Hcy demonstrate the largest affinity for silver, both in relation to methionine and cysteine residues in peptides. The studies have for the first time revealed the effect of predominant forming of ions containing two silver atoms per a sulfhydryl group of cysteine-bearing peptides if the nearest microenvironment does not hinder this.

[Ligands of glutamate and dopamine receptors evenly labeled with hydrogen isotopes].

A reaction of high-temperature solid-phase catalytic isotope exchange (HSCIE) was studied for the preparation of tritium- and deuterium-labeled ligands of glutamate and dopamine receptors. Tritium-labeled (5S,10R)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclopenten-5,10-imine ([G-(3)H]MK-801) and R(+)-7-hydroxy-N,N-di-n-propyl-2-aminotetraline ([G-(3)H]-7-OH-DPAT) were obtained with a specific activity of 210 and 120 Ci/mol, respectively. The isotopomeric distribution of deuterium-labeled ligands was studied using time-of-flight mass-spectrometer MX 5310 (ESI-o-TOF) with electrospray and orthogonal ion injection. Mean deuterium incorporation per ligand molecule was 11.09 and 3.21 atoms for [G-(2)H]MK-801 and [G-(2)H]-7-OH-DPAT, respectively. The isotope label was shown to be distributed all over the ligand molecule. The radioreceptor binding of tritium-labeled ligands [G-(3)H]MK-801 and [G-(3)H]-7-OH-DPAT was analyzed using the brain structure of Vistar rats. It was demonstrated that [G-(3)H]MK-801 specifically binds to hippocampus membranes with K(d) 8.3 +/- 1.4 nM, B(max) being 3345 +/- 300 fmol/mg protein. The [G-(3)H]-7-OH-DPAT ligand specifically binds to rat striatum membranes with K(d) 10.01 +/- 0.91 nM and B(max) 125 +/- 4.5 fmol/mg protein. It was concluded that the HSCIE reaction can be used for the preparation of highly tritium-labeled (+)-MK-801 and 7-OH-DPAT with retention of their physiological activities.

[On-drift spectrometer with double successive separation with ionization in the corona discharge].

An experimental high-resolution ion-drift spectrometer is described. Operating characteristics were improved by installation of a second grid gate in the center of the drift space. The ion-drift analyzer setups and the ranges of amplitude, length and temporary delay of control impulses were identified in the course of testing. The article gives illustration of mobility spectrum in case of a single or paired gate analysis.

[Arachidonoyl amino acids and arachidonoyl peptides: synthesis and properties].

N-Arachidonoyl (AA) derivatives of amino acids (glycine, phenylalanine, proline, valine, gamma-amino butyric acid (GABA), dihydroxyphenylalanine, tyrosine, tryptophan, and alanine) and peptides (Semax, MEHFPGP, and PGP) were synthesized in order to study the biological properties of acylamino acids. The mass spectra of all the compounds at atmospheric pressure electrospray ionization display the most intense peaks of protonated molecular ions; the detection limits for these compounds are 10 fmol per sample. AA-Gly showed the highest inhibitory activity toward fatty acid amide hydrolase from rat brain (IC50 6.5 microM) among all the acylamino acids studied. AA-Phe, AA-Tyr, and AA-GABA exhibited a weak but detectable inhibitory effect (IC50 55, 60, and 50 microM, respectively). The acylated amino acids themselves, except for AA-Gly, were stable to the hydrolysis by this enzyme. All the arachidonoylamino acids inhibited cabbage phospholipase D to various degrees; AA-GABA and AA-Phe proved to be the most active (IC50 20 and 27 microM, respectively). Attempts to detect the biosynthesis of AA-Tyr in homogenates of rat liver and nerve tissue showed no formation in vitro of either this acylamino acid or AA-dopamine and AA-Phe, the products of its metabolism. The highest contents of these metabolites were detected in liver homogenate and in the brain homogenate, respectively. Acylamino acids exert no cytotoxic effect toward the glioma C6 cells. It was shown that N-acylation of Semax with arachidonic acid results in enhancement of its hydrolytic stability and increases its affinity for the sites of specific binding in rat cerebellum membranes. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2006, vol. 32, no. 3; see also http://www.maik.ru.