Search results for "Isocratic elution"

Description of the Retention and Peak Profile for Chromolith Columns in Isocratic and Gradient Elution Using Mobile Phase Composition and Flow Rate a…

2014

The effect of the modifier concentration and flow rate on the chromatographic performance of a second generation Chromolith® RP-18e column, under isocratic and gradient elution with acetonitrile-water mixtures, was examined using four sulphonamides as probe compounds. The acetonitrile concentration was varied between 5 and 55% (v/v), and the flow rate between 0.1 and 5.0 mL/min, keeping the other factors constant. The changes in both retention and peak profile were modelled, and used to build simple plots, where the logarithm of the retention factor was represented against the modifier concentration (in gradient elution, against the initial modifier concentration), and the half-widths or wi…

Serial versus parallel columns using isocratic elution: a comparison of multi-column approaches in mono-dimensional liquid chromatography.

2015

Abstract When a new separation problem is faced with high-performance liquid chromatography (HPLC), the analysis is addressed conventionally with a single column, trying to find out a single experimental condition aimed to resolve all compounds. However, in practice, the system selectivity may be insufficient to achieve full resolution. When a separation fails, the usual practice consists of introducing drastic changes in the chromatographic system (e.g. use of another column, solvent or pH). An alternative solution is taking benefit of the combined separation capability of two or more columns, which can be attained in multiple ways, such as diverse modalities of two-dimensional HPLC, or mo…

An isocratic high-pressure liquid chromatographic purification method for radioactively labeled deoxyribonucleoside triphosphates.

1976

Abstract A method is described for the rapid purification of radioactively labeled deoxyribonucleoside tri­phosphates from their spontaneously emerging hydrolysis products deoxyribonucleoside diphosphate, deoxyribonucleoside monophosphate, and deoxyribonucleoside. The separations which are finished within 3 min or less are carried out on a 0.1X5 cm column filled with LiChrosorb-NH2 , using isocratic elution with 0.025 м potassium phosphate, pH 6 .8 , in a high-pressure liquid chromatograph at room temperature and a flow rate of 30 ml · h-1(flow velocity 63.7 cm·min-1).

Determination of hydroxytyrosol and tyrosol by liquid chromatography for the quality control of cosmetic products based on olive extracts

2014

An analytical method for the simultaneous determination of hydroxytyrosol and tyrosol in different types of olive extract raw materials and cosmetic cream samples has been developed. The determination was performed by liquid chromatography with UV spectrophotometric detection. Different chromatographic parameters, such as mobile phase pH and composition, oven temperature and different sample preparation variables were studied. The best chromatographic separation was obtained under the following conditions: C18 column set at 35°C and isocratic elution of a mixture ethanol: 1% acetic acid solution at pH 5 (5:95, v/v) as mobile phase pumped at 1 mL min(-1). The detection wavelength was set at …

Approaches to model the retention and peak profile in linear gradient reversed-phase liquid chromatography.

2012

Abstract The optimisation of the experimental conditions in gradient reversed-phase liquid chromatography requires reliable algorithms for the description of the retention and peak profile. As in isocratic elution, the linear relationship between the logarithm of the retention factor and the solvent contents is only acceptable in relatively small concentration ranges of modifier. However, more complex models may not allow an analytical integration of the general equation for gradient elution. Alternative approaches for modelling the retention in linear gradient elution are here proposed. Those based on the quadratic logarithmic model and a model proposed for normal liquid chromatography yie…

Effect of sodium dodecyl sulphate and Brij-35 on the analysis of sulphonamides in physiological samples using direct injection and acetonitrile gradi…

2016

Micellar liquid chromatography (MLC) is a reversed-phase (RP) mode, which often does not require gradient elution, since the peaks of mixtures of compounds within a large range of polarities appear more evenly distributed in the chromatograms of isocratic elution, giving rise to a “gradient effect”. However, the use of a gradient of organic solvent may still be convenient to shorten the total analysis time. This work compares the separation of 15 sulphonamides using conventional hydro-organic RPLC and MLC with the surfactants sodium dodecyl sulphate (SDS) and Brij-35, and both isocratic and gradient elution with acetonitrile. The observed behaviour is rationalized attending to the interacti…

Modeling of Retention in Reversed Phase Liquid Chromatography

2015

Half-width plots, a simple tool to predict peak shape, reveal column kinetics and characterise chromatographic columns in liquid chromatography: Stat…

2013

Peak profiles in chromatography are characterised by their height, position, width and asymmetry; the two latter depend on the values of the left and right peak half-widths. Simple correlations have been found between the peak half-widths and the retention times. The representation of such correlations has been called half-width plots. For isocratic elution, the plots are parabolic, although often, the parabolas can be approximated to straight-lines. The plots can be obtained with the half-widths/retention time data for a set of solutes experiencing the same kinetics, eluted with a mobile phase at fixed or varying composition. When the analysed solutes experience different resistance to mas…

Use of triacylglycerol profiles established by high performance liquid chromatography with ultraviolet–visible detection to predict the botanical ori…

2011

A method for the determination of triacylglycerols (TAGs) in vegetable oils from different botanical origins by HPLC with UV–vis detection has been developed. Using a core-shell particle packed column (C18, 2.6 micron), TAG separation was optimized in terms of mobile phase composition and column temperature. Using isocratic elution with acetonitrile/n-pentanol at 10 °C, excellent efficiency with good resolution between most of the TAG peak pairs, within a total analysis time of 15 min, was achieved. Using mass spectrometry detection, a total of 15 peaks, which were common to oils of six different botanical origins (corn, extra virgin olive, grapeseed, hazelnut, peanut and soybean) were iden…

Enhancement in the computation of gradient retention times in liquid chromatography using root-finding methods.

2019

Abstract Gradient elution may provide adequate separations within acceptably short times in a single run, by gradually increasing the elution speed. Similarly to isocratic elution, chromatograms can be predicted under any experimental condition, through strategies based on retention models. The most usual approach implies solving an integral equation (i.e., the fundamental equation of gradient elution), which has an analytical solution only for certain combinations of retention model and gradient programme. This limitation can be overcome by using numerical integration, which is a universal approach although at the cost of longer computation times. In this work, several alternatives to impr…