Biomedical subjects

Melvin R Euerby

Publications and source records attributed to Melvin R Euerby.

14 recordsLinked to original sources

PubMed2006

A study of retention and overloading of basic compounds with mixed-mode reversed-phase/cation-exchange columns in high performance liquid chromatography.

The retention and overload of bases were studied on two new mixed-mode, silica based phases possessing ionic carboxylate functionalities of different acidity embedded within a hydrophobic ligand (SiELC Primesep). At low pH, good peak shapes were obtained for small solute mass, suggesting that the mere presence of a mixed-mode hydrophobic/ionic retention mechanism is not responsible for the poor peak shape that can occur on conventional reversed-phases with ionised silanols. Somewhat inferior, but still acceptable peak shape for bases was obtained on a column containing a mixture of discrete ion exchange and reversed-phase particles (Hypersil Duet). In both types of column, the ionic sites favourably increased the capacity for ionised bases, reducing considerably the deterioration of peak shape with load observed with conventional RP columns. The combined ionic and reversed-phase interaction can give strong retention of bases under certain conditions, necessitating careful choice of stationary and mobile phase.

Chromatography, High Pressure Liquid↗

PubMed2006

Retention modelling in ternary solvent-strength gradient elution reversed-phase chromatography using 30 mm columns.

An optimization strategy for ternary solvent-strength gradient elution RP chromatography is described in which a two-dimensional model of gradient time (2 levels) against ternary proportions of organic modifiers (4 levels) was constructed. From the resolution surface the optimum ratio of organic modifiers could be selected. Excellent retention time and acceptable peak width and resolution simulations were obtained. The separation could be further optimized from the same input data by using a standard one-dimensional model in order to optimize for gradient slope, duration and shape. Excellent retention time and acceptable peak width and resolution simulations were obtained (< 1, 2 and 6% error, respectively).

Chromatography, High Pressure Liquid↗

PubMed2006

Study of overload for basic compounds in reversed-phase high performance liquid chromatography as a function of mobile phase pH.

The retention and overloading properties for eight basic solutes and two quaternary ammonium compounds were studied over the pH range 2.7-10.0 using phosphate and carbonate buffers. At low pH, a hybrid inorganic-organic silica-ODS phase (XTerra RP-18, 15 cm x 0.46 cm) showed substantial loss in efficiency when sample masses exceeded about 0.5 microg; these results were similar to those obtained previously on pure silica ODS and wholly polymeric phases, suggesting a common overloading mechanism. At pH 7-8.5, substantial improvements in loading capacity were obtained on XTerra due apparently to the unexpectedly strong influence of small decreases in solute ionisation. Data from the quaternary compounds suggested that silanol ionisation on this phase was still small even at intermediate pH. For many bases, loading capacity continued to improve as the pH was raised to 10, in line with the decrease in the proportion of ionised solute. However, for the highest pK(a) solutes, peak shape worsened at high pH, possibly due to the negative influence of increasing column silanol ionisation.

Amitriptyline↗

PubMed2006

The role of water in drug-receptor interactions.

The idea that liquid water is not a uniform and random arrangement of molecules has been taken very seriously by the scientific community. Many experimental and computational investigations show that clathrate- or ice-like structures probably exist at a short time scale in solution. We have designed a new program to simulate water structure around solutes. Our model is based on the geometrical constraints of hydrogen bonding in order to be capable of producing clathrate-like structures. Simulations with small molecules and bio-molecules, using the new software, produce networks of water with specific patterns made of small water rings. The water structures built are consistent with the classification of molecules in terms of structure breaking and making. This approach may give insight into, and a more accurate description of, drug-receptor interactions. The results also suggest that water structure may impart sufficient energy to modify the conformational space of organic molecules through hydrogen bonding.

Computer Simulation↗

PubMed2005

Chromatographic classification and comparison of commercially available reversed-phase liquid chromatographic columns containing polar embedded groups/amino endcappings using principal component analysis.

Polar embedded phases have become increasingly popular in liquid chromatography (LC) analysis. These phases can produce diverse chromatographic selectivities as a result of their differing base silica, the type of polar embedded group (i.e. amide, urea, carbamate, ether or sulphonamide moieties) and the length of the alkyl ligand. Four column characterization protocols, using differing test probes, have been used to characterize 18 of these phases together with 17 alkyl phases (some of which contained novel polar endcapping, i.e. amino), which have been evaluated using principal component analysis (PCA). PCA provided graphical comparisons of the differences/similarities between these phases and between their corresponding C-alkyl, amino endcapped and enhanced polar selectivity phases.

Amines↗

PubMed2005

Migration behavior of weakly retained, charged analytes in voltage-assisted micro-high performance liquid chromatography.

The application of voltage in micro-high performance liquid chromatography (micro-HPLC) creates a system where separation is governed by a hybrid differential migration process, which entails the features of both HPLC and capillary zone electrophoresis (CZE), i.e., chromatographic retention and electrophoretic migration. In this paper, we use our previously published approach to decouple these two mechanisms via analysis of the input data for estimation of electrokinetic parameters, such as conductivity, equivalent lengths, mobilities and velocities. Separation of weakly retained, charged analytes was performed via voltage-assisted micro-HPLC. Contrary to conclusions from data analysis using the conventional definitions of the retention factor, it is shown that our approach allows us to isolate the "chromatographic retention" component and thus, investigate the "modification" of the retention process upon application of voltage in micro-HPLC. It is shown that the traditional approaches of calculating retention factor would erroneously lead to conclusion that the retention behavior of these analytes changes upon application of voltage. However, the approach suggested here demonstrates that under the conditions investigated, most of the charged analytes do not show any significant retention on the columns and that all the changes in their retention times can be attributed to their electrophoretic migration.

Chromatography, High Pressure Liquid↗

PubMed2005

Capillary electrochromatography of therapeutic peptides on mixed-mode butylmethacrylate monoliths.

In this study, a porous mixed-mode n-alkyl methacrylate-based monolith has been used in the separation of therapeutic peptides. While the sulfonic acid (SCX) moiety derived from 2-acrylamido-2-methyl-1-propanesulfonic acid supports the generation of a stable electroosmotic flow (EOF) at both acidic and basic pH values, the butyl ligands provide the nonpolar sites for chromatographic resolution. The performance of the monolith was evaluated regarding the influence of pH on chromatographic resolution of peptides. The suitability of the butylmethacrylate/SCX monolith for the analysis of therapeutic peptides containing basic centres, for example arginine, at moderately high pH 9.5 and the stability to repeat injections of a mixture of peptides was demonstrated. Separations with efficiencies as high as 5.0 x 10(5) plates/m were obtained and the migration behaviour of the peptides at both low (2.8) and high (9.5) pH values could be rationalised based on their charge, molecular mass/shape and relative hydrophobicities.

Chromatography, High Pressure Liquid↗

PubMed2005

An evaluation of the robustness of the Tanaka characterization protocol for reversed-phase liquid chromatography columns.

The robustness of the popular 'Tanaka' LC column characterization protocol has been evaluated by the statistical tools of reduced factorial design, multiple linear regression and principal component analysis. These have shown that in order to obtain reliable and reproducible results, it is especially important to control the methanol content, the temperature and, in the case of the total ion-exchange capacity test (alphaB/P pH 7.6), the pH of the mobile phase. In particular, the hydrophobicity tests (kPB and alphaCH2) are sensitive to small changes in methanol content. Provided that the operating parameters for the Tanaka column characterization protocol are controlled within the following experimental limits, i.e. methanol content +/-0.5% v/v, temperature +/-3degreesC, pH +/-0.10 and buffer concentration +/-2.0 mM, it is feasible to distinguish between RP materials that possess selectivity differences larger than their batch-to-batch reproducibility. These experimental requirements can be easily met by current LC instrumentation. Hence, the validity of the Tanaka testing protocol for characterizing columns has been verified.

Benzene Derivatives↗

PubMed2004

Rationalisation of unusual changes in efficiency and retention with temperature shown for bases in reversed-phase high-performance liquid chromatography at intermediate pH.

Despite the possibility of poorer peak shapes, analysis of pharmaceuticals and other bases using reversed-phase high-performance liquid chromatography (RP-HPLC) at intermediate pH gives useful increases in retention, selectivity and column loading capacity compared with low pH. Retention times of some bases showed anomalous increases with column temperature. Peak shapes for bases improved significantly at elevated temperature (up to 70 degrees C), with the weakest bases studied (pKa approximately 8) giving greater improvement than the strongest bases (pKa approximately 10). In contrast, quaternary ammonium compounds showed reduced retention with increasing column temperature (normal behaviour) and only modest improvements in peak shape. Considering these results, and pKa measurements of the bases made using capillary electrophoresis, it appears that increases in retention and improvement in efficiency may be influenced significantly by reduction in the pKa of bases with temperature, leading to reduction in protonation. It is less likely that efficiency improvements are due to the speeding up of the kinetics of silanol ion exchange, at least in the temperature range studied here.

Chromatography, High Pressure Liquid↗

PubMed2004

Determination of ionisation constants of organic bases in aqueous methanol solutions using capillary electrophoresis.

The pKa of eight organic bases was determined in aqueous and aqueous methanol solutions of 0-70% (v/v) methanol using capillary electrophoresis. The bases investigated include compounds commonly used to test the activity of RP columns in HPLC. The variation of pKa with temperature in aqueous methanol solutions was also investigated and found to closely resemble temperature coefficients reported for bases in purely aqueous solutions. pKa values determined by CE were compared to those reported using NMR spectroscopy. The good agreement of the results is evidence that either technique is suitable to perform pKa measurements.

Acetonitriles↗

PubMed2003

Interplay of chromatographic and electrophoretic processes in capillary electrochromatography.

The separation mechanism in capillary electrochromatography (CEC) is a hybrid differential migration process, which entails the features of both high-performance liquid chromatography and capillary zone electrophoresis, i.e., chromatographic retention and electrophoretic migration. The adsorption of the different sample components on the stationary phase can be modified by the presence of the electric field across the column. Here, we use our previously published approach to decouple chromatographic retention from electrophoretic migration that allows us to investigate the "modification" of the retention process in CEC. This paper presents a methodology for characterization of changes in the retention of neutral and charged sample components, under identical conditions of stationary and mobile phase.

Adsorption↗

PubMed2003

Determination of pKa values of organic bases in aqueous acetonitrile solutions using capillary electrophoresis.

Capillary electrophoresis (CE) was used for the determination of ionisation constants (pKa) of a variety of organic bases in aqueous acetonitrile solutions over the range 0-60% (v/v) acetonitrile. These bases are used as test compounds in HPLC column evaluation, thus knowledge of their pKa in hydro-organic solutions is useful. The base pKa decreased with acetonitrile concentration and significant shifts from the aqueous pKa (up to -0.8) were found using 60% acetonitrile. The CE application was confirmed to be very suitable for fast and accurate pKa measurement in aqueous organic solutions.

Acetonitriles↗

PubMed2003

Chromatographic classification and comparison of commercially available reversed-phase liquid chromatographic columns using principal component analysis.

A total of 135 commercially available alkyl, cyano, phenyl, perfluorinated, polar embedded, enhanced polar selectivity (i.e., polar/hydrophilic endcapped), "Aqua type" and a variety of novel phases including some non-silica based stationary phases have been characterised in terms of their surface coverage, hydrophobic selectivity, shape selectivity, hydrogen bonding capacity and ion-exchange capacity at pH 2.7 and 7.6. Principal component analysis has been used to provide a simple graphical comparison of the differences/similarities between columns in the entire database and differing subsets such as "Aqua type"/enhanced polar selectivity phases. The PCA has been correlated to the phase's ability to analyse a range of hydrophilic bases.

Chromatography, Liquid↗

PubMed2002

Overloading study of bases using polymeric RP-HPLC columns as an aid to rationalization of overloading on silica-ODS phases.

The separation of ionized bases by reversed-phase liquid chromatography with alkyl silica columns often leads to severely tailed bands that are highly detrimental. Band shape and its dependence on sample mass are notably different when mobile-phase pH is changed, and this behavior has not been previously explained. Ionized silanols present in the stationary phase have been credited with a role in determining peak shape. In the present study, separations on two different polymer columns were compared with those previously obtained on alkyl silica phases. Because silanols are absent from polymer columns, this comparison enabled us to assess the role of silanols in separations on alkyl silica phases and to offer an explanation of why band shape changes with sample size and mobile-phase pH for both polymer and silica-based phases.

Journal Article↗