Prediction of the resolution of capillary columns in different conditions of inlet pressure and temperature

A procedure previously described for the prediction of the plate height of capillary columns operated at different inlet pressure of the carrier gas and at various column temperatures by using few retention data measured under isobaric conditions was modified and improved in order to permit the prediction of the retention times and of the peak widths at various heights. It is therefore possible to calculate the ratio, delta, between the peak width at different heights and the peak width at half height, whose value is used to predict the resolution at different height of two closely eluting peaks. It was found that the delta values do not depend on temperature and inlet pressure and are a characteristic of the used column; they can therefore be used in order to calculate the resolution in any temperature and inlet pressure condition. The method was used to predict the retention time, the peak width and the resolution of polar and non-polar compounds (alkanes, alkenes, chloroalkanes, alcohols, ketones) on capillary columns of different length and polarity by using as the starting data retention and width values measured in three isobaric runs only.     

The use of predicted boiling points for the identification of halobenzenes and substituted phenols in capillary gas chromatography

Quantitative correlations between physicochemical parameters and structure of various solutes and their gas chromatographic behavior were investigated in order to predict the retention values. The identification of unknown samples in gas chromatographic routine analysis of environmental samples is enhanced by the use of these correlations in conjuction with other chromatographic methods and with mass spectrometry. The boiling points of many compounds are easily found in literature and therefore their correlation with retention values obtained in isothermal or programmed temperature analysis permit restriction of the choice of names of unknown compound to within a narrow range. The correlation between the boiling points and the retention times of chloro- and bromo-benzenes and of some chloro- and nitro- substituted phenols was investigated on non-polar capillary columns and permitted the tentative identification of many compounds belonging to these homologous series.     

Prediction of retention values in linear temperature programming of narrow and mega-bore capillary columns

Retention times in linear temperature programmed gas chromatography on narrow bore and megabore capillary columns have been calculated from experimental retention times measured at three isothermal temperatures: an iterative procedure performed by BASIC programs was used to obtain the values of the constants enabling the calculation of the programmed temperature retention times. Different methods of calculation have been compared.     

Comparison of mathemical methods for the calculation of retention indices at high temperature in gas chromatography

The retention indices of three homologous series (2-alkanones, 1-alkanols, cycloalksanones) have been determined at high temperature by the application of two new adaptation methods: A multiparametric least-squares regressions iterative method based on the dertermination of the adjusted retention times and a cubic interpolation directly using the uncorrected retention times without dead time correction. The two methods were applied to two types of columns. The first group includes eight packed columns (seven OV polymethylphenylsiloxane and Apolane-87 stationary phases), while the second includes five glass capillary columns (four methyl-silicons with different film thicknesses and Apolane-87 stationary phases). The retention indices obtained with a multiparametric and a cubic interpolation methods were compared with each other and with those calculated by Grobler's, Guardino's, Kaiser's and Kovàts' methods. The influence of coating, film thickness, and temperature on them was investigated.     

A general strategy for performing temperature-programming in high performance liquid chromatography--prediction of segmented temperature gradients

In the present work it is shown that the linear elution strength (LES) model which was adapted from temperature-programming gas chromatography (GC) can also be employed to predict retention times for segmented-temperature gradients based on temperature-gradient input data in liquid chromatography (LC) with high accuracy. The LES model assumes that retention times for isothermal separations can be predicted based on two temperature gradients and is employed to calculate the retention factor of an analyte when changing the start temperature of the temperature gradient. In this study it was investigated whether this approach can also be employed in LC. It was shown that this approximation cannot be transferred to temperature-programmed LC where a temperature range from 60°C up to 180°C is investigated. Major relative errors up to 169.6% were observed for isothermal retention factor predictions. In order to predict retention times for temperature gradients with different start temperatures in LC, another relationship is required to describe the influence of temperature on retention. Therefore, retention times for isothermal separations based on isothermal input runs were predicted using a plot of the natural logarithm of the retention factor vs. the inverse temperature and a plot of the natural logarithm of the retention factor vs. temperature. It could be shown that a plot of lnk vs. T yields more reliable isothermal/isocratic retention time predictions than a plot of lnk vs. 1/T which is usually employed. Hence, in order to predict retention times for temperature-gradients with different start temperatures in LC, two temperature gradient and two isothermal measurements have been employed. In this case, retention times can be predicted with a maximal relative error of 5.5% (average relative error: 2.9%). In comparison, if the start temperature of the simulated temperature gradient is equal to the start temperature of the input data, only two temperature-gradient measurements are required. Under these conditions, retention times can be predicted with a maximal relative error of 4.3% (average relative error: 2.2%). As an example, the systematic method development for an isothermal as well as a temperature gradient separation of selected sulfonamides by means of the adapted LES model is demonstrated using a pure water mobile phase. Both methods are compared and it is shown that the temperature-gradient separation provides some advantages over the isothermal separation in terms of limits of detection and analysis time.     

Comparing columns for gas chromatography with the two-parameter model for retention prediction

The retention times of selected compounds in temperature programmed gas chromatography were predicted using a two-parameter model, on the basis of thermodynamic data obtained from isothermal runs on seven capillary columns, primarily substituted with 5% diphenylsiloxane. The scope for using thermodynamic data obtained from isothermal runs on one column to optimize separation on a different column or a different instrument setup was investigated. Additionally, the predictive utility of thermodynamic data obtained using a DB-5 column that had been in use for three years was compared to that of a new column of the same model. It was found that satisfactory separation could be achieved on one capillary column or instrument setup on the basis of thermodynamic data obtained using a different column or instrument set-up.     

Evaluation of the Linear-Elution-Strength Model for the Prediction of Retention and Selectivity in Isothermal Gas Chromatography

Linear-elution strength theory and temperature-programmed gas chromatography is evaluated as a rapid method for predicting isothermal retention factors and column selectivity. Retention times for a wide range of compounds are determined at the program rates of 3 and 12 °C/min for the temperature range 60 to 160 °C on three open-tubular columns (DB-1701, DB-210 and EC-Wax) and used to predict isothermal retention factors for each column over the temperature range 60 to 140 °C. The temperature-program predicted isothermal retention factors are compared with experimental values using linear regression and the solvation parameter model. It is shown that isothermal retention factors predicted by the linear-elution-strength model only approximately represents the experimental data. The model fails to predict the slight curvature that exists in most plots of the experimental retention factor (log k) as a function of temperature. In addition, regression of the temperature-program predicted isothermal retention factors against the experimental values indicates that the slopes and intercepts deviate significantly from their target values of one and zero, respectively, in a manner which is temperature dependent. The temperature-program predicted isothermal retention factors result in system constants for the solvation parameter model that are different to those obtained from the experimental retention factors. These results are interpreted as indicating that linear-elution-strength theory predicts retention factors that fail to accurately model stationary phase interactions over a wide temperature range. It is concluded that temperature-program methods using linear-elution-strength theory are unsuitable for constructing system maps for isothermal separations.     

Temperature programmed retention indices: Calculation from isothermal data. Part 1: Theory

Direct conversion of isothermal to temperature programmed indices is not possible. In this work it is shown that linear temperature programmed retention indices can only be calculated from isothermal retention data if the temperature dependence of both the distribution coefficients and the column dead time are taken into account. Procedures are described which allow calculation of retention temperatures and from these, accurate programmed retention indices. Within certain limits the initial oven temperature and programming rate can be chosen freely. The prerequisite for this calculation is the availability of reliable isothermal retention data (retention times, retention factors, relative retention times, or retention indices) at two different temperatures for one column. The use of compiled isothermal retention indices at two different temperatures for the calculation of retention temperatures and thus temperature programmed indices is demonstrated. For the column for which programmed retention indices have to be determined, the isothermal retention times of the n-alkanes and the column dead time as a function of temperature have to be known in addition to the compiled data for a given stationary phase. Once the programmed retention indices have been calculated for a given column the concept allows the calculation of temperature programmed indices for columns with different specifications. The characteristics which can be varied are: column length, column inner diameter, phase-ratio, initial oven temperature, and programming rate.     

Determination of thermodynamic characteristics from retention data in GC

Summary Retention data have been measured for model substances of different polarities under isothermal conditions at several temperatures using capillary columns coated with Carbowax-20M and SP-2100 stationary phases. By using the k capacity ratios obtained for various temperatures the thermodynamic characteristics of the partition process were determined. For homologous series linear relationships can be established between thermodynamic characteristics and carbon number. Thermodynamic characteristics can be used both to compare and evaluate column quality and to predict retention indices for different temperatures and linear programmed temperature conditions.Dedicated to Professor J. F. K. Huber on the occasion of his 60th birthday.     

Prediction of the separation number of capillary columns in programmed temperature gas chromatographic analysis

The efficiency of capillary columns in programmed temperature analysis can be evaluated by calculation of the separation number (“Trennzahl”). A procedure for the prediction of this parameter at various initial temperatures, carrier gas pressures and heating rates, by using as the starting data the retention times and the peak widths obtained in some isobaric and isothermal runs is described. An equation is proposed that permits to obtain the values of the peak width at half height in any isothermal and linearly programmed temperature gas chromatographic run and therefore to calculate the separation number value. The effect on this parameter of the column polarity was investigated by using polar and non-polar compounds (n-alkanes and 1-alcohols).     

Prediction of retention times and peak widths in temperature-programmed gas chromatography using the finite element method

Optimization of separations in gas chromatography is often a time-consuming task. However, computer simulations of chromatographic experiments may greatly reduce the time required. In this study, the finite element method was used to predict the retention times and peak widths of three analytes eluting from each of four columns during chromatographic separations with two temperature programs. The data acquired were displayed in predicted chromatograms that were then compared to experimentally acquired chromatograms. The differences between the predicted and measured retention times were typically less than 0.1%, although the experimental peak widths were typically 10% larger than expected from the idealized calculations. Input data for the retention and peak dispersion calculations were obtained from isothermal experiments, and converted to thermodynamic parameters.     

Retention models for programmed gas chromatography

The models proposed by many authors for the prediction of retention times and temperatures, peak widths, retention indices and separation numbers in programmed temperature and pressure gas chromatography by starting from preliminary measurements of the retention in isothermal and isobaric conditions are reviewed. Several articles showing the correlation between retention data and thermodynamic parameters and the determination of the optimum programming rate are reported. The columns of different polarity used for the experimental measurement and the main equations, mathematical models and calculation procedures are listed. An empirical approach was used in the early models, followed by the application of thermodynamic considerations, iterative calculation procedures and statistical methods, based on increased computing power now available. Multiple column arrangements, simultaneous temperature and pressure programming, applications of two-dimensional and fast chromatography are summarised.     

Extrapolation and interpolation of gas chromatographic retention times

Summary An empirical method of extrapolating and interpolating gas chromatographic retention times obtained at three equally spaced isothermal temperatures is described. The accuracy of the method was evaluated from retention time data obtained using packed glass columns. A procedure for constructing retention time tables for homologous series and the derivation of an equation for calculating retention times as a function of temperature is also presented.     

Analysis of the equations for the temperature dependence of the retention index. I. Relation between equations.

The relation between the parameters of several equations for the retention index temperature dependence was established, taking the hyperbola deduced from the retention theory as starting point. The transformation factors depend only on methylene contributions to the thermodynamic functions of solution and temperature. Their evolution with the mean temperature of the range was illustrated for SE-30 and Carbowax-20M glass capillary columns. On this basis the post-run standardisation of dI/dT values at a reference mean temperature is possible. Examples and statistical correlation between series of parameters from different equations for perfumery solutes were shown.     

Use of Retention Data as the First Step in the Identification of Cyclic Organic Peroxides in Temperature-Programmed Gas Chromatography

Temperature-programmed retention indices for eleven cyclic organic peroxides were determined by gas chromatography on slightly polar 5% biphenyl 95% dimethylpolysiloxane columns (DB-5 and Rtx-5MS) at three heating rates (5, 10, and 20° min⁻¹) from 60 to 300°C, using different chromatographs. Cyclic organic diperoxides and triperoxides had nearly constant retention indices when different heating rates and a short isothermal hold time (5 min) before the programmed increase in temperature were used. The usefulness of temperature-programmed retention indices was shown by using the data to predict the retention times and structures of unknown diperoxides or triperoxides derived from ketones. This is the first step in the identification of unknown cyclic organic peroxides, a process would otherwise require the availability of reference compounds. Revised: 7 and 17 November 2005     

An homologous series of benzodiazepine retention index standards for gas chromatography

An homologous series of benzodiazepine retention index standards (the R-series) has been synthesized and the gas chromatographic behavior of the series investigated on NB-54 and NB-1701 capillary columns. The compounds were stable, exhibited symmetrical peak shapes, and fairly linear retention behavior was observed on both columns. The series can be coinjected with every sample to enable the high precision analysis of toxicological samples; screening for 20 benzodiazepine drugs was possible in 23 minutes (including cooling). The R-series method was compared with a retention index method based on a series of benzodiazepine drugs as standards and with a method employing relative retention times. The precision of the R-series method was found to be generally better than that of the two other methods in both long- and short-term studies.     

Modelling the gas chromatographic separation of multicomponent samples on a system of three open-tubular columns coupled in series

A procedure was developed for modelling the gas chromatographic separation on a system of columns coupled in series. This procedure can be used for computer based optimization of lengths and order of serially coupled columns at isothermal conditions. The sample component retention factors and column resistances needed for the model can easily be measured on each individual column. The proposed procedure was verified by analyzing a 52 component hydrocarbon mixture on three columns of different polarity coupled in series by various column orders. Good agreement between experimental and calculated retention data was found. The procedure is also well suited for optimization of the chromatographic selectivity by coupling columns with different selectivity in series.Dedicated to Professor Dr. h.c. mult. J.F.K. Huber on the occasion of his 70th birthday     

The impact of column inner diameter on chromatographic performance in temperature gradient liquid chromatography

The impact of column inner diameter on chromatographic performance in temperature gradient liquid chromatography has been investigated in the present study. Columns with inner diameters of 0.32, 0.53, 3.2 and 4.6 mm were compared with respect to retention and efficiency characteristics using temperature gradients from 30 to 90 degrees C with temperature ramps of 1, 5, 10 and 20 degrees C min(-1). The columns were all of 15 cm length and were packed with 3 microm Hypersil ODS particles. Alkylbenzenes served as model compounds, and the mobile phase consisted of acetonitrile-water (50:50, v/v). The study revealed that the column ID is not a critical limiting factor when performing temperature programming in LC, at least for columns narrower than 4.6 mm inner diameter in the temperature interval 30-90 degrees C. The retention times for all components on all columns were highly comparable, with similar peak profiles without any signs of peak splitting. The use of mobile phase pre-heating when using the larger bore columns was avoided by starting the temperature gradients close to ambient. However, the relative apparent efficiency was inversely proportional to column inner diameter, making the capillary columns generally more functional towards temperature gradients than the larger bore columns with respect to chromatographic efficiency. In addition, the capillary columns possessed higher robustness towards temperature programming than the conventional columns.     

Band acceleration device for enhanced selectivity with tandem-column gas chromatography

An electrically heated and air cooled metal sheath surrounding the first 50 cm of the second column in a series-coupled, capillary-column ensemble of a non-polar and a polar column is used to obtain enhanced isothermal separation of component pairs that are separated by the first column in the ensemble but co-elute from the ensemble by virtue of the different selectivity of the two columns. As the first of the two components passes into the second column, a current pulsed through the metal sheath rapidly heats the first 50 cm of the second column thus accelerating the band for the first component. Ensemble retention-time shifts of several seconds are easily obtained. The device is then rapidly cooled to quiescent oven temperature by a flow of pressured air through the space between the metal sheath and the fused silica capillary column and an additional flow through a larger, co-axial plastic tube. Both heating and cooling require only a few seconds. If substantial cooling of the device occurs before the band for the second component enters the device, the band experiences less thermally-induced acceleration with the result that the separation of the two targeted components is enhanced in the ensemble chromatogram with no significant change in the pattern of peaks for the other mixture components. If the device is cooled to a temperature below oven temperature before the arrival of the band for the second component, this band will be slowed, and further enhancement of separation is achieved in the ensemble chromatogram. A band trajectory model, based on retention factor versus temperature data for the two components in the two columns, is used to predict peak separation and to aid in the selection of temperature-pulse initiation times.