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Biomedical subjects

B G Belenkii

Publications and source records attributed to B G Belenkii.

9 recordsLinked to original sources

Modified capillary electrophoresis system for peptide, protein and double-stranded DNA analysis.

The results of high-performance capillary electrophoresis (HPCE) studies of peptide, protein and double-stranded DNA separations on a laboratory-made HPCE system are presented. Parameters of the HPCE system are given. The new method of capillary surface modification by grafting poly(glycidyl methacrylate) is described. The problems of HPCE biopolymer analysis connected with the sample-wall interactions are discussed.

DNA↗

High-performance membrane chromatography.

In gradient chromatography for proteins migrating along the chromatographic column, the critical distance X0 has been shown to exist at which the separation of zones is at a maximum and band spreading is at a minimum. With steep gradients and small elution velocity, the column length may be reduced to the level of membrane thickness--about one millimeter. The peculiarities of this novel separation method for proteins, high-performance membrane chromatography (HPMC), are discussed and stepwise elution is shown to be especially effective. HPMC combines the advantages of membrane technology and high-performance liquid chromatography, and avoids their drawbacks.

Chromatography, High Pressure Liquid↗

Peculiarities of zone migration and band broadening in gradient reversed-phase high-performance liquid chromatography of proteins with respect to membrane chromatography.

The peculiarities of zone migration and band broadening in the reversed-phase gradient HPLC of proteins were investigated. In the isocratic mode a critical composition of the mobile phase was found at which all proteins regardless of their molecular mass migrate with equal velocity and have a capacity factor equal to the phase ratio (VP/V0), i.e., the same capacity factor as a marker of total accessible volume would have in steric exclusion chromatography. It is shown that steric exclusion conditions are never achieved in gradient HPLC. In the first (adsorption stage) of gradient elution where the separation takes place the velocity of a protein increases until it becomes equal to the velocity of the desorbing solvent front at a critical distance X0 from column entrance. Strong broadening is characteristic of this stage. In the second (critical) stage the protein travels the remaining distance (L-X0) with the velocity of the solvent. A definition of X0 is given allowing one very simple calculation of the minimum permissible column length as a function of gradient steepness, mobile phase velocity and protein adsorption parameter. When x = X0 the protein zone has the smallest dispersion. Making L < X0 is especially disadvantageous, as it leads to anomalous bandspreading. The theory of gradient HPLC was refined on this basis and the usefulness of this approach in high-performance membrane chromatography is demonstrated.

Adsorption↗

Review. Use of laser detectors in capillary liquid chromatography.

The main relationship of high-performance liquid chromatography (HPLC) are considered. It is shown that the optimum conditions of ultrasensitive trace analysis should be achieved by using packed capillary columns manufactured from flexible quartz capillaries with dc approximately less than 0.2 mm. The main features of these columns (v opt = 0.6 v opt of that for conventional HPLC columns with double the hydraulic permeability) make it possible to obtain two or three times higher plate numbers for the same analysis time and column pressure characteristic of conventional HPLC, as a result of using a submicrometre sorbent. The main features of laser detection in capillary liquid chromatography (laser-induced fluorescence and cross-beam thermal lens absorption detectors) are considered. The requirements that should be met by a modern capillary liquid chromatograph based on using flexible quartz capillary columns with a submicrometre sorbent and laser detectors are formulated. Examples of using these systems for femtomole and attomole analyses of biological samples (amino acids and prostaglandins) are given.

Chromatography, High Pressure Liquid↗

Determination of myoglobin in human serum by high-performance liquid chromatography with chemiluminescence detection.

A highly sensitive method for the determination of myoglobin in serum is described, based on high-performance size-exclusion chromatography with chemiluminescence detection. Serum proteins are separated according to their molecular masses on columns packed with TSK-SW gel and those containing haem are detected selectively by post-column chemiluminescence reaction with luminol using a conventional fluorimetric detector. The method is rapid (30 min) and sufficiently sensitive for the diagnosis of myocardial infarction. The minimum detectable myoglobin concentration is 10 ng/ml.

Chromatography, High Pressure Liquid↗

Use of combined chromatographic methods including thin-layer chromatography for analysis of complex polymer systems. Determination of the polydispersity of block copolymers of styrene and methyl methacrylate by gel permeation, thin-layer and pyrolysis gas chromatography.

A combination of gel permeation chromatography (GPC), thin-layer chromatography (TLC) and pyrolysis gas chromatography (PGC) has been used for investigations of a polymethyl methacrylate-polystyrene-polymethyl methacrylate block copolymer. Continuous distribution of the polymer (40-mg sample) was attained according to the content of the styrene and methyl methacrylate units and of the block copolymer and according to the composition of the copolymer as functions of the hydrodynamic radius of the macromolecules. The polymer was subjected to a preliminary fractionation with an analytical gel chromatograph. The fractions were investigated by TLC, which permitted the separation of the block copolymer and the homopolymers. The composition of the fractions obtained by GPC and TLC was determined by PGC. As a result, it was possible to establish the composition of the block copolymer and its ratio to polymethyl methacrylate in each fraction. This investigation was based on a combination of highly effective fractionation by chromatographic methods with precise quantitative ratios obtained from Benoit's universal calibration graph and from determinations of the composition of the polymer fractions by PGC. The mechanism of the TLC of polymers, including the appearance of artefacts that distort the results of analysis, is also discussed.

Chromatography↗