An overview on the standardization of chromatographic methods for screening analysis in toxicology by means of retention indices and secondary standards

Summary A review is presented on the methods of standardization of HPLC data as used in systematic toxicological analysis. In straight-phase HPLC, the best results were obtained with a series of selected drugs as retention standards. In reversed-phase HPLC, various retention index systems were introduced. However, these systems alone cannot compensate large differences in selectivities of nominally identical, but commercially different reversed-phase column packings. Much better results were achieved with selected drugs as retention index markers. The practical applicability of such a standardized HPLC system is demonstrated.Part I see [1]Dedicated to Prof. Dr. Dr. Marika Geldmacher-von Mallinckrodt in recognition of her pioneering contributions in clinical and forensic toxicology     

一种提高色谱指纹谱保留时间重现性的新方法

通过色谱热力学分析发现，在相同的分析条件下，即使采用不同的液相色谱系统或不同的色谱柱，组分的保留时间存在简单的线性关系，应用该线性关系可提高不同反相C18柱间保留时间重现性，经过实际样品在不同操作条件下的验证，表明该方法是正确而可行的。     

Reduction of silanophilic interactions in liquid chromatography with the use of ionic liquids

A suppression of silanophilic interactions by the selected ionic liquids added to the mobile phase in thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) is reported. Acetonitrile was used as the eluent, alone or with various concentrations of water and phosphoric buffer pH 3. Selectivity of the normal (NP) and the reversed (RP) stationary phase material was examined using a series of proton-acceptor basic drugs analytes. The ionic liquids studied appeared to significantly affect analyte retention in NP-TLC, RP-TLC and RP-HPLC systems tested. Consequently, the increased separation selectivity was attained. Due to ionic liquid additives to eluent even analytes could be chromatographed, which were not eluted from the silica-based stationary phase materials with 100% of acetonitrile in the mobile phase. Addition of ionic liquid already in very small concentration (0.5%, v/v) could reduce the amount of acetonitrile used during the optimization of basic analytes separations in TLC and HPLC systems. Moreover, the influence of temperature on the separation of basic analytes was demonstrated and considered in practical HPLC method development.     

High-performance liquid chromatography and supercritical fluid chromatography of monosaccharides and polyols using light-scattering detection Chemometric studies of the retentions

Twelve sugars and polyols were analyzed using high-performance liquid chromatographic (HPLC) and supercritical fluid chromatographic (SFC) systems with silica and bonded silica stationary phases with the help of an evaporative light-scattering detector. The separation capacities of the two techniques are discussed. The retention data were studied using different chemometric methods (automatic classification, factor analysis). They clearly show that SFC and HPLC have the same retention process for these compounds and that is the sum of only two mathematically independent physico-chemical phenomena.     

Collection of analytical data for benzodiazepines and benzophenones

On April 1st, 1986, amendments were made to Schedule 2 to the Misuse of Drugs Act 1971 such that 33 benzodiazepines became controlled as Class C drugs in the U.K. An analytical database has been prepared to aid the detection and identification of controlled benzodiazepines. Chromatographic properties have been measured including gas chromatography (GC) retention index values, high-performance liquid chromatography (HPLC) capacity factors and thin-layer chromatography (TLC) RF x 100 values. UV spectroscopic and mass spectrometric (MS) data have also been recorded. Analytical data (GC, TLC and MS) are also presented for benzophenones which were produced by acid hydrolysis of the parent benzodiazepines.     

Overpressured layer chromatography in comparison with thin-layer and high-performance liquid chromatography for the determination of coumarins with reference to the composition of the mobile phase

The retention behaviour of fifteen closely related coumarins in normal-phase overpressured layer chromatography (OPLC) was studied with the aim of comparing the retentions with those in normal-phase thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) when optimization of the mobile phase was carried out according to the PRISMA system. The mobile phase optimization was carried out on TLC plates in unsaturated chambers. The resulting mobile phases were transposed to off-line, non-equilibrated OPLC and further to HPLC. The retention in TLC was measured at 37 selectivity points and in OPLC and HPLC at 13 points. Capacity factors (k′) and separation factors () were calculated in order to study the retention behaviour in the different systems. Two- and three-dimensional evaluations of k′ against selectivity points showed similar retention behaviours for the coumarins in TLC, OPLC and HPLC. The values for TLC, OPLC and HPLC showed similar patterns in the three-dimensional evaluations. The retention behaviour at different solvent strengths was also examined. According to quadratic regression, k′ showed a dependence on the change in solvent strength. OPLC, which can be considered as a “planar column” technique, and TLC are closely related methods, whereas HPLC shows a different behaviour in the elution process with regard to solvent strength.     

Use of SE-30 as a stationary phase for the gas-liquid chromatography of drugs.

The dimethyl silicone elastomer SE-30 has been chosen as the preferred liquid phase for the gas-liquid chromatographic analysis of drugs, and retention index data have been compiled for 480 drugs and commonly occurring chemicals such as plasticisers. The inter-laboratory variation in measurement of retention indices has been measured for three drugs in eleven laboratories and the standard deviations were between 20 and 15 retention index units.     

High-performance liquid chromatography systems for the separation of local anaesthetic drugs with applicability to the analysis of illicit cocaine samples

Two high-performance liquid chromatography (HPLC) systems are presented which are suitable for the separation of local anaesthetic drugs. An octadecyl-silica column is used with aqueous methanolic eluents (15 and 50% respectively) containing n-hexylamine-orthophosphoric acid buffers and data for 36 compounds are given. The HPLC systems have application for the identification of unknown drugs in this class while the first eluent (15% methanol, pH 2.5) is particularly useful for the examination of illicit cocaine samples because it gives good separation of common adulterants and impurities.     

Quantitative structure chromatography relationships in reversed-phase high performance liquid chromatography: Prediction of retention behaviour using theoretically derived molecular properties

Summary The use of theoretically calculated molecular properties as predictors for retention in reversed-phase HPLC has been explored. HPLC retention times have been measured for a series of 47 substituted aromatic molecules in three solvent mixtures and steric and electronic properties of these compounds have been derived using semi-empirical molecular orbital and empirical theoretical methods. A subset of the experimental data (a training set) was used to derive property-retention time relationships and the remaining data were then used to test the predictive capability of the methods.Good retention time prediction was possible using derived regression equations for individual solvents and after including solvent parameters it was possible to predict retention for all solvents using a single equation. This method showed that the most useful properties were calculated log P and the calculated dipole moment of the solutes, and the calculated solvent polarisability. In addition, 90% of the data were used to train an artificial neural network and the remaining 10% of the data used to test the network; excellent prediction was obtained, the neural network approach being as successful as the regression analysis.     

Selection of orthogonal reversed-phase HPLC systems by univariate and auto-associative multivariate regression trees

In order to select chromatographic starting conditions to be optimized during further method development of the separation of a given mixture, so-called generic orthogonal chromatographic systems could be explored in parallel. In this paper the use of univariate and multivariate regression trees (MRT) was studied to define the most orthogonal subset from a given set of chromatographic systems. Two data sets were considered, which contain the retention data of 68 structurally diversive drugs on sets of 32 and 38 chromatographic systems, respectively. For both the univariate and multivariate approaches no other data but the measured retention factors are needed to build the decision trees. Since multivariate regression trees are used in an unsupervised way, they are called auto-associative multivariate regression trees (AAMRT). For all decision trees used, a variable importance list of the predictor variables can be derived. It was concluded that based on these ranked lists, both for univariate and multivariate regression trees, a selection of the most orthogonal systems from a given set of systems can be obtained in a user-friendly and fast way.     

Oil-in-Water Microemulsion High Performance Liquid Chromatographic Analysis of Pharmaceuticals

Efficient and novel oil-in-water microemulsion HPLC separations of a range of solutes have been achieved using isocratic elution and a conventional reversed-phase HPLC column. A single set of isocratic oil-in-water microemulsion HPLC conditions are shown to be successful for the resolution of a wide range of basic, neutral and acidic drugs and excipients. A robust separation method was developed for the quantitative analysis of Naproxen in a tablet formulation. The method offered a similar run time and improved chromatography when compared to conventional HPLC modes, demonstrating its potential for routine use. In developing the separations the effect on the chromatography of varying the operating parameters was studied. While generally the method was robust to changes in some variables, others markedly changed the separation selectivity, solute retention, peak-peak resolution or resulted in poor chromatographic performance. The method was found to be compatible with very low UV working detection wavelengths. As a final stage to the experimental programme, a microemulsion method was developed with the capability of analysing a range of water-insoluble drugs.     

Retention behaviour of selected flavonoid compounds in NP-HPLC

Chromatographia - HPLC retention data for a number of flavonoid compounds have been evaluated on normal phase silica and cyano colums according to the Soczewinski-Snyder (S.-S.) model. On the...     

Determination of poly(ethylene glycol)s by both normal-phase and reversed-phase modes of high-performance liquid chromatography

A normal-phase HPLC system using an amino column has been developed to characterise oligomers of poly(ethylene glycol)s (PEGs) of average M_r 400 to 2000 with derivatisation by dinitrobenzoate. Normal-phase HPLC with gradient elution using ternary solvents of hexane, dichloromethane and methanol has produced a baseline resolution for oligomers of PEG 400, 600 and 1000, while PEG 1000 and 2000 were analysed by using binary solvents of acetonitrile and water. Mixtures of PEGs have been determined by these HPLC systems. PEG 400 in a textile finish has also been determined with satisfactory recovery. It has been found that the hydroxyl group of solvents in normal-phase HPLC plays an important role in resolution and retention of PEG oligomers. Derivatisation efficiency for PEGs by dinitrobenzoyl chloride and quantitative determination of derivatised PEGs by HPLC have been studied. A reversed-phase (RP) mode of HPLC was examined for determination of PEG 400 oligomers. The normal-phase system provided greater resolution for oligomers of PEGs.