The contribution of the pressure drop to analyte retention in coupled column supercritical fluid chromatography of lipids

Summary This paper reports the qualitative and quantitative effects of the column pressure drop on the retention of lipid components in a serially coupled capillary column SFC system. The contribution of the pressure drop consists of two components, the density effect and the flow effect. The magnitude of the flow effect,i. e. the change in retention which results from changes in the flow-rate when column pressures are changed, is determined by the difference in single column analyte k values. The effect will be positive compared with the uncorrected retention values when the column with largest k value is closest to the injector. With the columns in reversed order, the effect will be negative. The contribution from the density effect always resulted in larger coupled column k values and was in most instances of more significance than the flow effect component. Values calculated with and without pressure drop correction have been compared and it has been shown that for most of the eighteen model lipid compounds investigated, the deviations from the experimental retention factors were smaller when pressure drop corrections were made.     

Usage of nonporous polymeric particles for reversed-phase high-performance liquid chromatography of oxidized and deamidated forms of recombinant human growth hormone

In this work, a C(18) reversed-phase column with nonporous polymeric 2.5- micro m particles is utilized to initially test the analysis of oxidized and deamidated human growth hormone (hGH). Phosphate buffer (pH 7.5) with 24% 1-propanol was used for elution. This quick method (analysis time is 20 min) gave a selectivity, as judged by the number of detected peaks, and resolution of hGH variants that is better than many methods in which porous silica particle columns are used. Only mixtures of oxidized and deamidated hGH are analyzed, and no characterization of the peaks is performed. The results indicate that C(18) nonporous polymeric column material is a promising alternative for the chromatographic separation of several hGH variants.     

Solid-phase extraction and gas chromatographic-mass spectrometric identification of degradation products from enhanced environmentally degradable polyethylene

A solid-phase extraction (SPE) method using unbonded silica (Si) and silica bonded with octadecyl (C₁₈) or aminopropyl (NH₂) groups was developed to separate into five fractions the highly complex mixture of low-molecular-mass degradation products formed from degradable polymers. Application of the method to polyethylene modified with starch and/or a pro-oxidant system, degraded for 30 weeks in water at 95°C, enabled the identification by GC-MS of over three times as many products as when the sample was prepared by liquid-liquid extraction. Over 60 degradation products were identified in each sample; mainly dicarboxylic acids, monocarboxylic acids and n-alkanes. In addition, several lactones, aldehydes and alcohols were detected.     

The roles of template complexation and ligand binding conditions on recognition in bupivacaine molecularly imprinted polymers

A model for the molecular basis for ligand recognition in bupivacaine imprinted methacrylic acid-ethylene glycol dimethacrylate co-polymers has been developed based upon a series of (1)H-NMR studies in conjunction with HPLC and radioligand binding analyses. (1)H-NMR studies indicated that functional monomer-template complexes survive the polymerisation process, at least up until the stage of gelation. Polymers were synthesised and characterised by surface area analysis (BET), FT-IR and SEM. A combination of zonal and frontal chromatographic studies in aqueous and non-polar media indicate that selectivity arises from a combination of hydrophobic and electrostatic interactions. However, in the concentration regime employed for LC-based studies, ligand recognition in aqueous media was shown to be predominantly non-specific and hydrophobic in character. Radioligand binding studies, in lower ligand binding concentration regimes, permitted closer examination of the higher affinity binding sites. It was shown that the presence of a polar modifier in a non-polar solvent, or an organic modifier in water, produced enhanced selectivity. Variable temperature studies showed that the temperature of binding influences selectivity as well as the apparent number of sites available and that this effect is different in organic and aqueous environments. This indicates that the system studied is more complex in character than is generally appreciated. A comparison of the techniques employed here indicates that although chromatographic studies provide a valuable first-round screen for polymer-ligand selectivities, the level of detail obtainable using radioligand binding studies (lower concentrations and true equilibrium binding) makes them superior for detailed evaluations of molecularly imprinted polymers.     

The spin turning in ferromagnetic Gd studied by positive muons

We have studied the muon precession frequency in a ferromagnetic single crystal of Gd metal. The overall features of our findings are compatible with earlier results on polycrystalline material. In the temperature region between 245 and 220 K where the Gd magnetization starts to turn away from the c-axis, we observe an increase in the muon depolarization rate, and a complex precession signal which can be separated into two frequency components meaning that spin turning does not occur simultaneously in different parts of the sample (domains). From these more detailed data follows that previously obtained values forB _fc andB _dip can not both be correct. Two explanations for our new result are possible: EitherB _fc undergoes a change around 230 K which is directly coupled to the spin turning angle, or the value of the dipolar field contribution used in the earlier evaluation is too low. This imposes some uncertainty as to the value of the angle at the onset of spin turning derived fromSR frequencies.     

Double photoionisation spectra of small molecules and a new empirical rule for double ionisation energies

Complete double photoelectron spectra are presented for 18 small molecules where the location of charges in the cations and dications is relatively clearly defined. The data demonstrate the importance of a coulombic repulsion contribution to the double ionisation energies. Examination of data for a wide range of molecules leads to a new empirical rule to calculate double ionisation energies from the molecules’ single ionisation energies and maximum dimensions. Where single and double ionisation energies are known the rule allows the deduction of plausible intercharge distances.     

Optimisation of chiral separation of omeprazole and one of its metabolites on immobilized α1-acid glycoprotein using chemometricsglycoprotein using chemometrics

Summary A strategy for the optimisation of direct chiral separation of omeprazole and a metabolite, hydroxi-omeprazole, in reversed phase liquid chromatography is described. A factorial design was used, where mobile phase pH, concentration of a mobile phase modifier, ionic strength and column temperature were tested as the variables and enantioselective retention, column efficiency and asymmetry factor as the responses. The experimental results were evaluated with multivariate analyses, which demonstrated that the column temperature and content of mobile phase acetonitrile were by far the most important variables. The enantiomers of omeprazole and one of its metabolites were baseline resolved within 15 minutes. The optimised chromatographic system was used for a separation of the enantiomers of omeprazole and its main metabolite in a patient plasma sample.