Biomedical subjects

Salwa K Poole

Publications and source records attributed to Salwa K Poole.

5 recordsLinked to original sources

PubMed2004

Automated robotic liquid handling/laser-based nephelometry system for high throughput measurement of kinetic aqueous solubility.

The ability to rapidly and consistently measure aqueous solubility in a preclinical environment is critical to the successful identification of promising discovery compounds. The advantage of an early solubility screen is timely attrition of compounds likely to fail due to poor absorption or low bioavailability before more costly screens are performed. However, due to the large number of compounds and limited sample amounts, thermodynamic solubility measurements are not feasible at this stage. A kinetic solubility measurement is an alternative to thermodynamic measurements at the discovery stage that provides a rank listing of solubility values with minimal sample requirements. A kinetic solubility measurement is attractive from an automation vantage because it features rapid data acquisition and is amenable to multi-well formats. We describe the use of a robotic liquid/plate handler coupled to nephelometry detection for the measurement of kinetic solubility. We highlight the liquid handling validation, serial dilution parameters, and a comparison to the previous method. Experiments to further enhance throughput, or increase confidence in the automation steps, are described and the effects of these experiments are presented. In our integrated nephelometry method, we observe rapid liquid handling with an error of less than 10%, after a series of validation studies, and a sample throughput up to 1800 compounds per week. We compare the nephelometry method with our semi-thermodynamic flow-injection analysis (FIA) method, and find a 75% bin agreement between the methods.

Kinetics↗

PubMed2004

Determination of acid dissociation constants by capillary electrophoresis.

Capillary electrophoresis affords a simple, automated approach for the measurement of pKa values in the range 2-11 at a throughput of less than 1 h per sample per instrument. Agreement with literature values is usually within 0.20 log units with a precision better than 0.07 log units. The attractive features of capillary electrophoresis for pKa measurements are: (1) conventional instrumentation with a high level of automation are suitable for all measurements; (2) because it is a separation method samples need not be of high purity; (3) samples of low water solubility with suitable chromophores are easily handled (detection limits in the microM range); (4) sample consumption per measurement is in the microgram range; and (5) since only mobilities are measured, exact knowledge of concentrations is not needed. The general approach can be extended to pKa measurements in aqueous-organic solvent mixtures and non-aqueous solvents with suitable calibration. The widespread use of absorbance detection in capillary electrophoresis means that the sample must have a suitable chromophore for detection. The main source of controllable error is the accuracy of buffer standardization and their stability in use, and uncontrollable error, the retentive interactions of the sample with the column wall. The latter seems to be a rare problem in practice for typical operating conditions.

Acids↗

PubMed2003

Separation methods for estimating octanol-water partition coefficients.

Separation methods for the indirect estimation of the octanol-water partition coefficient (logP) are reviewed with an emphasis on high throughput methods with a wide application range. The solvation parameter model is used to identify suitable separation systems for estimating logP in an efficient manner that negates the need for empirical trial and error experiments. With a few exceptions, systems based on reversed-phase chromatography employing chemically bonded phases are shown to be unsuitable for estimating logP for compounds of diverse structure. This is because the fundamental properties responsible for chromatographic retention tend to be different to those responsible for partition between octanol and water, especially the contribution from hydrogen bonding interactions. On the other hand, retention in several micellar and microemulsion electrokinetic chromatography systems is shown to be highly correlated with the octanol-water partition coefficient. These systems are suitable for the rapid, high throughput determination of logP for neutral, weakly acidic, and weakly basic compounds. For compounds with a permanent charge, electrophoretic migration and electrostatic interactions with the stationary phase results in inaccurate estimation of partition coefficients. The experimental determination of solute descriptors offers an alternative approach for estimating logP, and other biopartitioning properties. A distinct advantage of this approach is that once the solute descriptors are known, solute properties can be estimated for any distribution or transport system for which a solvation parameter model has been established.

Chromatography, Liquid↗

PubMed2003

Estimation of octanol-water partition coefficients for neutral and weakly acidic compounds by microemulsion electrokinetic chromatography using dynamically coated capillary columns.

Microemulsion electrokinetic chromatography (MEEKC) using dynamically coated capillary columns is shown to be suitable for estimating the octanol-water partition coefficient (log P) for neutral and weakly acidic compounds at pH 3. The solvation parameter model is used to demonstrate that the retention properties of sodium dodecyl sulfate (1.4% w/v), n-butanol (8% v/v) and n-heptane (1.2% v/v) microemulsion are strongly correlated with the octanol-water partition system. For compounds of varied structure and log P values from 0.3 to 5.15, the correlation model is able to estimate log P to better than 0.25 log units. The dynamically coated columns consisting of a bilayer of poly(vinylsulfonate) adsorbed on top of polybrene provide a suitable electroosmotic flow at pH 3 without interfering in the retention properties of the microemulsion. For automated measurements the microemulsion run buffer should be replenished after 10 runs to maintain a stable cycle time and the coated columns replaced after 40-70 runs, depending on sample properties.

Acids↗

PubMed2002

Column selectivity from the perspective of the solvation parameter model.

The solvation parameter model is a useful tool for delineating the contribution of defined intermolecular interactions to retention of neutral molecules in separation systems based on a solute equilibrium between a gas, liquid or fluid mobile phase and a liquid or solid stationary phase. The free energy for this process is decomposed into contributions for cavity formation and the set up of intermolecular interactions identified as dispersion, electron lone pair, dipole-type and hydrogen bonding. The relative contribution of these interactions is indicated by a series of system constants determined by the difference of the defined interaction in the two phases. The interpretation of these system constants as a function of experimental factors that affect retention in the chromatographic system provides the connection between relative retention (selectivity) and the control variables for the separation system. To aid in the understanding of these processes we perform an analysis of system constants for gas chromatography, liquid chromatography, supercritical fluid chromatography and micellar electrokinetic chromatography as a function of different experimental variables as a step towards gaining a theoretical understanding of selectivity optimization for method development.

Chromatography, Gas↗