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The analysis of amino acids presents significant challenges to contemporary analytical separations. The present paper investigates the possibility of retention prediction in hydrophilic interaction chromatography (HILIC) gradient elution based on the analytical solution of the fundamental equation of the multilinear gradient elution derived for reversed-phase systems. A simple linear dependence of the logarithm of the solute retention (ln k) upon the volume fraction of organic modifier (φ) in a binary aqueous-organic mobile is adopted. Utility of the developed methodology was tested on the separation of a mixture of 21 amino acids carried out with 14 different gradient elution programs (from simple linear to multilinear and curved shaped) using ternary eluents in which a mixture of methanol and water (1:1, v/v) was the strong eluting member and acetonitrile was the weak solvent. Starting from at least two gradient runs, the prediction of solute retention obtained under all the rest gradients was excellent, even when curved gradient profiles were used. Development of such methodologies can be of great interest for a wide range of applications., (Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The use of turbulent flow chromatography (TFC) as a method for the rapid metabonomic LC-MS analysis of plasma as an alternative to solvent-based protein precipitation has been investigated. This comparison has shown that TFC can be effectively used in this application with the benefit that off-line sample handling is significantly reduced. However, analysis of the data obtained via TFC for human plasma reveals substantial differences in the overall metabolite profiles compared with methanol-precipitated HPLC-MS. This seems in part at least to be related to greatly reduced amounts of phospholipids (ca. 10 fold reduction) for the turbulent flow methodology compared with protein-precipitated samples. The significance of these differences with respect to metabolite profiles as a result of the sample preparation method used are discussed.

Hydrophilic interaction chromatography (HILIC) is a relatively recently introduced mode of liquid-phase separations. Recently, HILIC has been used for coupling to MS in metabonomic/metabolomic studies to provide a complementary tool to the widely used reversed-phase (RP) chromatographic separations. The combination of HILIC with MS detection covers a number of polar metabolites that are typically nonretained in RPLC-mode separations and thus enlarging the number of detected analytes. This way of metabolite profiling thus provides more comprehensive metabolome coverage than using RP chromatography alone. This review describes the applications and the utility of HILIC-MS in metabolomic/metabonomic studies and highlights certain characteristic examples in the life and plant-food sciences.

The determination of widely used anaesthetic and analgesic drugs in biological fluids is of major clinical importance. Typical methods used for sample preparation employ liquid-liquid extraction protocols which are complex, costly, not handy and not amenable to automation. In the present communication, we report the development of a methodology that employs headspace solid-phase microextraction (HS-SPME) for the determination of four anaesthetic (lidocaine, midazolam, diazepam and ketamine) and three analgesic drugs (fentanyl, remifentanyl and codeine) in human urine. Important parameters controlling SPME were studied: selection of SPME fibre, type and amount of salt added, preheating and extraction time, extraction temperature, sample volume and desorption time. GC with nitrogen phosphorus detection (GC-NPD) facilitates sensitive and selective detection of the anaesthetics. The developed method renders an efficient tool for the precise and sensitive determination of the anaesthetics and analgesics in human urine (RSDs ranged from 7.7 to 12.6%, whereas LODs ranged from 0.01 to 1.5 ng/mL). The method was applied to the determination of the anaesthetics and analgesics in human urine from patients that had undergone coronary by-pass surgery operations. The proposed protocol can function as an attractive alternative for clinical acute intoxications and medico-legal cases.

A new capillary electrophoretic method for the chiral separation of four major Cinchona alkaloids (quinine/quinidine and cinchonine/cinchonidine) was developed using heptakis-(2,6-di-O-methyl)-beta-cyclodextrin as the chiral selector. The inner walls of the separation capillary were modified with a thin polyacrylamide layer, which substantially reduced the electroosmotic flow and improved the chiral resolution and the reproducibility of the migration time of the analytes. Various operation parameters were optimised, including the pH, the capillary temperature, the concentration of the background electrolyte, and the concentration of the chiral selector. Baseline separation of the two diastereomer pairs was achieved in 12 minutes in ammonium acetate background electrolyte pH 5.0 with addition of cyclodextrin in a concentration of 3 mM or higher.

As shown in the past years, SPE based on molecularly imprinted polymers (MIPs) may provide significant enhancement of selectivity in sample preparation and analyte preconcentration. The objective of this work was the fabrication of MIPs for the specific adsorption of rutin and quercetin. The two flavonoids were used as the template molecules for the preparation of MIP phases in a self-assembly (noncovalent) approach. The produced MIPs were validated with regard to the imprinting efficiency as media for LC and SPE. The retention behavior of several flavonoid compounds was studied using as stationary phases imprinted, control nonimprinted polymers, and commercial silica-based materials. MIPs were applied as materials for the selective SPE and preconcentration of the flavonoids from white and red wine, orange juice, and tea. The collected fractions were analyzed by high-pressure LC. MIP-SPE facilitated specific analyte isolation and effective sample clean-up. The results show that molecularly imprinted SPE can be a useful tool for the simple, selective, and cost-effective pretreatment of samples containing natural antioxidants.