Pressurized capillary electrochromatographic assay of trimethoprim impurities using 1 microm particle-based columns

One micrometre silica particles, derivatized with C18, were electrokinetically packed into a 75-microm-i.d. capillary. The resulting column was evaluated for the separation of trimethoprim (TMP) and its impurities using pressurized capillary electrochromatography (pCEC), starting from a capillary liquid chromatographic (CLC) separation. These samples require gradient elution when separated by high performance liquid chromatography (HPLC), but with the new columns isocratic elution suffices for their separation by CLC or pCEC. Only 70,000 theoretical plates/m for impurity C were achieved using CLC mode at relative low pressure (78 bar) although very small particles were utilized. When a voltage above 2 kV (50 V/cm) was applied, unknown peaks appeared, which was assumed due to an electrophoretic effect with the unknown peaks resolving as a result of the applied voltage. In order to minimize these unfavorable contributions, only a low voltage was applied, still leading to higher separation performances and shorter separation times than in CLC. The optimal analyzing conditions in pCEC included a pressure of 78 bar, an applied voltage of 1 kV, and a mobile phase consisting of 80 mM sodium perchlorate (pH 3.1)/methanol (60/40, v/v). These conditions were used to separate and quantify four major impurities in TMP within 22 min. The obtained calibration curves were linear (r>0.9980) in concentration ranges between 0.005 and 0.1 mg/mL for impurities A and C; 0.02 and 0.10 mg/mL for impurity F; and 0.01 and 0.10 mg/mL for impurity H. The detection limits (S/N=3) for impurities A, C, F, and H were 0.52, 0.84, 3.18, and 2.41 microg/mL, respectively. The calibration curves were successfully applied to analyze spiked bulk samples, with mean recoveries ranging from 92% to 110%. The developed method can therefore be considered simple, rapid, and repeatable.     

Modelling the quality of enantiomeric separations using Mutual Information as an alternative variable selection technique

This paper uses Mutual Information as an alternative variable selection method for quantitative structure-property relationships data. To evaluate the performance of this criterion, the enantioselectivity of 67 molecules, in three different chiral stationary phases, is modelled. Partial Least Squares together with three commonly used variable selection techniques was evaluated and then compared with the results obtained when using Mutual Information together with Support Vector Machines. The results show not only that variable selection is a necessary step in quantitative structure-property relationship modelling, but also that Mutual Information associated with Support Vector Machines is a valuable alternative to Partial Least Squares together with correlation between the explanatory and the response variables or Genetic Algorithms. This study also demonstrates that by producing models that use a rather small set of variables the interpretation can be also be improved.     

Enantiomeric impurity determination in capillary electrophoresis using a highly-sulfated cyclodextrins-based method

Capillary electrophoresis (CE), using highly-sulfated cyclodextrins as chiral selectors, has been applied to determine the chiral purity of pharmaceutical compounds. A chiral separation strategy, developed earlier for racaemic mixtures, was applied on four basic drugs (propranolol, atenolol, chlorpheniramine and tryptophan methylester). The aim was to develop validated separation methods which allow determination of 0.1% impurity levels of the unwanted enantiomers (distomer) in the presence of 99.9% of the active compound (eutomer). The linearity, quantification limits for the trace enantiomers and the precision of the measurements were determined. In a second part, impurity separations have been simulated in order to evaluate the required resolution when assaying impurities. It is shown that a baseline resolution of 1.5, generally accepted for racaemic mixtures, does not always allow good impurity determinations. Two alternative methods to solve this problem have been proposed.     

Interface, Surface Tension and Reentrant Pinning Transition in the 2D Ising Model

We develop a new way to look at the high-temperature representation of the Ising model up to the critical temperature and obtain a number of interesting consequences. In the two-dimensional case, it is possible to use these tools to prove results on phase-separation lines in the whole phase-coexistence regime, by way of a duality transformation. We illustrate the power of these techniques by studying an Ising model with a boundary magnetic field, in which a reentrant pinning transition takes place; more precisely we show that the typical configurations of the model can be described, at the macroscopic level, by interfaces which are solutions of the corresponding thermodynamic variational problem; this variational problem is solved explicitly. There exist values of the boundary magnetic field and temperatures 0<T ₁<T ₂<T _c such that the interface is not pinned for T<T ₁ or T>T ₂, but is pinned for T ₁<T<T ₂; we can also find values of the boundary magnetic field and temperatures 0<T ₁<T ₂<T ₃<T _c such that for T<T ₁ or T ₂<T<T ₃ the interface is pinned, while for T ₁<T<T ₂ or T>T ₃ it is not pinned. An important property of the surface tension which is used in this paper is the sharp triangle inequality about which we report some new results. The techniques used in this work are robust and can be used in a variety of different situations. Received: 28 January 1998 / Accepted: 22 December 1998     

Shotgun Proteomics of Isolated Urinary Extracellular Vesicles for Investigating Respiratory Impedance in Healthy Preschoolers

Urine proteomic applications in children suggested their potential in discriminating between healthy subjects from those with respiratory diseases. The aim of the current study was to combine protein fractionation, by urinary extracellular vesicle isolation, and proteomics analysis in order to establish whether different patterns of respiratory impedance in healthy preschoolers can be characterized from a protein fingerprint. Twenty-one 3–5-yr-old healthy children, representative of 66 recruited subjects, were selected: 12 late preterm (LP) and 9 full-term (T) born. Children underwent measurement of respiratory impedance through Forced Oscillation Technique (FOT) and no significant differences between LP and T were found. Unbiased clustering, based on proteomic signatures, stratified three groups of children (A, B, C) with significantly different patterns of respiratory impedance, which was slightly worse in group A than in groups B and C. Six proteins (Tripeptidyl peptidase I (TPP1), Cubilin (CUBN), SerpinA4, SerpinF1, Thy-1 membrane glycoprotein (THY1) and Angiopoietin-related protein 2 (ANGPTL2)) were identified in order to type the membership of subjects to the three groups. The differential levels of the six proteins in groups A, B and C suggest that proteomic-based profiles of urinary fractionated exosomes could represent a link between respiratory impedance and underlying biological profiles in healthy preschool children.     

A simple cladding process to apply monolithic silica rods in high performance liquid chromatography

A simple cladding procedure was developed to apply monolithic silica rods in chromatography. This was used to evaluate the performances of new monolithic silica phases synthesised using a triblock copolymer as a phase separation inducer. The cladding procedure is found efficient after several tests, including scanning electron microscopy and mercury porosimetry, and the obtained columns present a relatively good efficiency in adsorption chromatography (H min is about 50 μm) as compared to other lab-scale developed columns.     

Kulinkovich-type reactions of thioamides: similar to those of carboxylic amides?

The behaviour of thioamides under Kulinkovich-type conditions is compared with the known reactivity of carboxylic amides. Dramatic differences are disclosed.