Coupling of chiral and achiral stationary phases in supercritical fluid chromatography: Evaluating and improving retention prediction.

Isomers and stereoisomers are always challenging to separate. Column coupling may provide improved chromatographic selectivity, necessary for the separation of the compounds with similar chemical and structural properties. The relatively low viscosity of supercritical fluids, used as mobile phases allows for the coupling of several columns in series in supercritical fluid chromatography (SFC), without exceeding the pressure limits of the system. The aim of this study is to propose reliable prediction of the retention behaviour of analytes on a coupled column system, based on a limited number of initial analyses. The chiral compounds atenolol, ephedrine, propranolol, mianserin, labetalol and nadolol, besides the diastereomers quinine and quinidine, and the structural isomers of aminophenol and aminocresol were used as model analytes. The retention behaviour of the analytes was determined on the individual chiral columns Lux Cellulose-1, Lux Cellulose-2, Lux Cellulose-3, Lux Cellulose-4, Lux Amylose-2 and the achiral columns Luna NH2, Luna Silica, Synergi RP and FluoroSep RP. The mobile phase was composed of CO2 mixed with 20% (v/v) MeOH, which contained 0.1% (v/v) trifluoroacetic acid and 0.1% (v/v) isopropylamine. The retention factors of the analytes on coupled stationary phases were predicted, and subsequently compared to the experimentally obtained ones. Relative deviations of predicted and experimental retention factors were in range from 0.00% to 51.91%. Flow rate and back pressure of the screening conditions were adjusted to improve prediction precision on four column combinations, with varying success rates. The average relative deviations of retention factors were reduced to 2.84% - 6.59% by adjusting flow rate, and to 2.30% - 8.57% by adjusting back pressure. The most successful approach, flow rate adjustment, was then applied to select a column combination providing improved resolution of the structurally similar components of silymarin extract.

In-line molecularly imprinted polymer solid phase extraction-capillary electrophoresis coupled with tandem mass spectrometry for the determination of patulin in apple-based food.

We present a simple and sensitive method for the determination of patulin at µg·kg-1 level in apple-based products. Our method relies on the application of an in-line molecularly imprinted polymer solid-phase extraction microcartridge in capillary electrophoresis coupled with mass spectrometry. Capillary zone electrophoresis method has been developed and parameters affecting the in-line process have been carefully optimized. Validation parameters were assessed for patulin, giving LOQ of 1 µg·kg-1 and linearity range 1-100 µg·kg-1 with R2 ≥ 0.997. The LOQ was below the maximum content of patulin requested by the European Union in this type of products. The precision of the peak area and the migration time were less than 14.9 and 1.6%, respectively. Patulin has been analyzed in the presence of 5-hydroxymethylfurfural, which is the main interference in this kind of matrix. The method was applied to assay patulin content in various apple-based products.