A systematic investigation of algorithm impact in preparative chromatography with experimental verifications.

Computer-assisted optimization of chromatographic separations requires finding the numerical solution of the Equilibrium-Dispersive (ED) mass balance equation. Furthermore, the competitive adsorption isotherms needed for optimization are often estimated numerically using the inverse method that also solves the ED equations. This means that the accuracy of the estimated adsorption isotherm parameters explicitly depends on the numerical accuracy of the algorithm that is used to solve the ED equations.

Potential of adsorption isotherm measurements for closer elucidating of binding in chiral liquid chromatographic phase systems.

The human body is a chiral environment and many drugs are chiral and interact differently depending on the type of enantiomer. Therefore, the interest in analytical and preparative separations of enantiomers has steadily increased over the years. LC is today the most important technique in analytical laboratories worldwide.

A quest for the optimal additive in chiral preparative chromatography.

Traditionally, the choice of acid/base additives used in chiral preparative chromatography has not been considered very important. However, it was recently demonstrated that strongly adsorbing additives can result in the most unexpected enantiomer band shapes in modern chiral preparative chromatographic systems. In the present study we demonstrate that, depending on the choice of additive, it is actually possible to obtain the following four binary band-shape compositions when a racemic mixture is injected: (i) anti-Langmuir/anti-Langmuir, (ii) anti-Langmuir/Langmuir, (iii) Langmuir/Langmuir and (iv) Langmuir/anti-Langmuir.

Approach for reliable evaluation of drug proteins interactions using surface plasmon resonance technology.

The surface plasmon resonance (SPR) biosensor was recently introduced to the analytical biochemical society for measuring small drug-protein interactions. However, the technique has many times been used without specifying the type of enantiomeric form of the chiral drug measured and/or with using a too narrow drug concentration range resulting in biased values of binding coefficients and sometimes even assumptions about single-site bindings although the binding in reality comprises a multisite interaction. In this study we will give guidelines for reliable experimental and methodological approaches to avoid these pitfalls.

Adsorption behaviour of a quinidine carbamate-based chiral stationary phase: role of the additive.

In this study, we incorporate the additive properties into the theoretical model of a general preparative chromatographic system; this is normally not done and this limits a proper process optimization. As a model phase system, we used the adsorption of 9H-fluoren-9-ylmethoxycarbonyl-allylglycine (Fmoc-allylglycine) enantiomers on a quinidine carbamate-based chiral stationary phase (anion exchanger) together with a methanol-glacial acetic acid-ammonium acetate eluent. The inverse method was used to measure the competitive adsorption isotherms of both the Fmoc-allylglycine enantiomers as well as the non-detectable additive acetic acid.

Development of the tracer-pulse method for adsorption studies of analyte mixtures in liquid chromatography utilizing mass spectrometric detection.

The tracer-pulse method provides the real adsorption data points directly from simple, straightforward calculations and is therefore a superior method for multicomponent adsorption isotherm determination in HPLC. Only one important problem has restricted its use so far: the tracer peaks are invisible using any conventional detection principle. We present a solution to this problem with an approach with a firm base in analytical chemistry, utilizing stable isotopes and mass spectrometric detection.

Biotechnological approach to the synthesis of 9alpha-hydroxylated steroids.

The steroid 9alpha-hydroxylase gene has been cloned from Mycobacterium smegmatis into Escherichia coli BL21. Progesterone added to bioreactors was subjected to in vivo transformation into 9alpha-hydroxyprogesterone. In 7 days, 43.

Tuneable peak deformations in chiral liquid chromatography.

Modern chiral stationary phases are often combined with eluents comprising a mixture of organic solvents and polar additives. The latter may cause extreme deformations of the eluted enantiomer bands in both analytical and preparative separations. In this work, we give a theoretical background for these deformations.

Validation of the tracer-pulse method for multicomponent liquid chromatography, a classical paradox revisited.

The tracer-pulse method was extended and validated for the determination of multicomponent adsorption isotherms in liquid chromatography. Competitive adsorption isotherms can be determined for any number of solutes, up to the column resolution limit. The basic principle is to equilibrate the column with an eluent containing a mixture of the solutes and then measure the migration velocity of each of them through the column.

Invisible analyte peak deformations in single-component liquid chromatography.

It is well known that if a small excess of solute is injected into a chromatographic system equilibrated with an eluent containing the same solute, a single so-called perturbation peak will appear in the chromatogram. It was recently shown (Samuelsson, J.; Forssén, P.

Analytical characterization of chiral drug-protein interactions: comparison between the optical biosensor (surface plasmon resonance) assay and the HPLC perturbation method.

Two modern, fundamentally different methods were used for a detailed investigation of enantioselective drug-protein interactions, a surface plasmon resonance (SPR)-based Biacore 2000 biosensor assay and the previously validated HPLC perturbation method (HPLC-PM). This is the first time SPR has been used for this purpose. The fundamental features of the two methods were investigated, and the consequences for operation and data evaluation were addressed.

Accurate and rapid estimation of adsorption isotherms in liquid chromatography using the inverse method on plateaus.

The inverse method (IM) is an attractive approach for estimating adsorption isotherm parameters in liquid chromatography (LC), mainly due to its experimental simplicity and low sample consumption. This article presents a new experimental approach, the inverse method on plateaus (IMP), which uses elution profiles on concentration plateaus together with IM. This approach enabled us to obtain very accurate adsorption isotherms that agreed well with those estimated by means of frontal analysis over the entire concentration range under consideration.