P. Campíns-Falcó

Column-switching techniques for high-performance liquid chromatography of drugs in biological samples

In recent years, an increasing number of publications have demonstrated the potential of column-switching techniques for the chromatographic separation, determination and preparative isolation of analytes from biological matrices. Column-switching systems greatly facilitate drug analysis, by on-line sample clean-up and trace enrichment, or by improving the analytical separative process. In this paper, the main applications of column-switching techniques to drug analysis in biological samples, are reviewed.

Evaluation and elimination of the blank bias error using the H-point standard addition method : application to spectrophotometric determinations using absorbent blank

Abstract The basis of the H-point standard addition method in the cases where the analyte determination requires the use of an absorbent reagent is considered. The method evaluates and eliminates the blank bias error present in such procedures by using the absorbance increment at two selected wavelengths as the analytical signal of the calibration graphs. Three different determinations were tested: the determination of proteins with the biuret method, thorium with thoron and of magnesium with Titan Yellow. These procedures are better described by the proposed method than by the traditional approach using absorbance values against reagent blank.

Automatic in-tube SPME and fast liquid chromatography: A cost-effective method for the estimation of dibuthyl and di-2-ethylhexyl phthalates in environmental water samples

A 80-cm length commercially available capillary coated with 95% polydimethylsiloxane and 5% polydiphenylsiloxane (TBR-5) was employed to carry out on-line extraction and preconcentration of dibuthyl phthalate (DBP) and di-2-ethylhexyl phthalate (DEHP) in the chromatographic system. The coated capillary was placed between the sample injection loop and the injection needle of an autosampler. Variables affecting the automatic in-tube solid-phase microextraction (SPME) were optimized. A Genesis C(18) (5 cm x 4.6 mm i.d., 4 microm particle size) was employed as analytical column. The achieved limits of detection by use of diode array detection were 1 and 2.5 microg L(-1), respectively. The proposed conditions have been applied to determine those compounds at low ppb levels (< or =250 microg L(-1)) in aqueous samples. No matrix effect was found, and recoveries between 85 and 115% were obtained. The precision of the method was good, and the achieved intra- and inter-day variation coefficients were between 5 and 20%. The analysis time per sample was 20 min and any off-line pre-treatment of the samples was needed. The taken sample volume was 100 microL. Data on the application of the described method to the analysis of different water samples are presented.

New micromethod combining miniaturized matrix solid-phase dispersion and in-tube in-valve solid-phase microextraction for estimating polycyclic aromatic hydrocarbons in bivalves.

Miniaturized matrix solid-phase dispersion (MSPD) was developed for the extraction of common polycyclic aromatic hydrocarbons (PAHs) from bivalve samples (100mg, dry weight). Additional clean-up and analyte enrichment was accomplished by in-tube solid-phase microextraction (SPME). For this purpose the extracts collected after MSPD were diluted with water and injected into a capillary column coated with the extractive phase. This capillary column was connected to the analytical column by means of a switching valve. Separation and quantification of the PAHs were carried out using a monolithic LC column and fluorescence detection. Since the in-tube SPME device allowed the processing of large volumes of the extracts (2.0 mL) excellent sensitivity was achieved, thus making solvent evaporation operations unnecessary. The overall recoveries ranged from 10% to 28% for the studied compounds. The relative standard deviation (RSD) ranged from 2% to 10% for intra-day variation (n=3), and the limits of detection (LODs) were < or =0.6 ng/g (dry weight). The proposed procedure was very simple and rapid (total analysis time was approximately 20 min), and the consumption of organic solvents and extractive phases was drastically reduced. The reliability of the proposed MSPD/in-tube SPME method was tested by analysing several bivalves (mussels and tellins) as well as a standard reference material (SRM).

Analysis of 18 perfluorinated compounds in river waters: Comparison of high performance liquid chromatography–tandem mass spectrometry, ultra-high-performance liquid chromatography–tandem mass spectrometry and capillary liquid chromatography–mass spectrometry ☆

In this work, the performance of ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) and capillary liquid chromatography-mass spectrometry (CLC-MS) has been studied for the analysis of eighteen perfluorinated compounds in water samples. UHPLC-MS/MS and CLC-MS analysis were carried out using a Zorbax C-18 column (50 mm × 2.1 mm, 1.8 μm) and a Zorbax SB-C18 column (150 mm × 0.5 mm, 3.5 μm), respectively, in gradient elution mode with a mobile phase of ammonium formate and methanol. Both techniques were compared with conventional LC-MS/MS in terms of speed, sensitivity, selectivity and resolution. Water samples were extracted by solid phase extraction (SPE). Mean absolute recoveries at two concentration levels, 6 and 60 ng L⁻¹, calculated from the eighteen compounds were: 77 ± 12(s) and 82 ± 12(s), respectively. Relative standard deviation (RSD) was lower than 16% for each perfluorinated compound. The results obtained showed that UHPLC-MS/MS and CLC-MS allow the determination of perfluorinated compounds in water samples with satisfactory sensitivity and resolution and reduced analysis time. Therefore, both techniques were employed for the analysis of water samples to determine the target analytes at low concentration levels and with lower analysis times than using HPLC-MS/MS. Similar statistical values were obtained from both techniques at 95% confidence level. Several compounds have been detected, but perfluorooctanoic acid (PFOA) (171 ng L⁻¹ and 148 ng L⁻¹ by UHPLC-MS/MS and CLC-MS, respectively) showed the highest concentration.

Urine polyamines determination using dansyl chloride derivatization in solid-phase extraction cartridges and HPLC

The derivatization of biogenic amines such as putrescine, cadaverine, spermidine and spermine with dansyl chloride in solid phase extraction cartridges is described. Different types of filling materials were tested in order to have the highest retention of the different analytes. The best results were obtained by using C18 cartridges. The optimal conditions were: amine solution buffered at pH 12, 2 mM dansyl chloride (acetone-bicarbonate solution 20 mM (pH 9-9.5), 2 + 3 v/v) as reagent concentration, room temperature and 30 min reaction time. The developed procedure was applied to the determination of these polyamines in urine samples from healthy controls and cancer patients using HPLC with 1,7-diaminoheptane as internal standard. The concentrations ranged from 0.5 to 5 micrograms mL-1 and the detection limits were 10 ng mL-1 for all polyamines. By concentrating the urine extracts, the detection limits were improved down to 2 ng mL-1. The accuracy and the precision of the method were tested. The proposed dansylation method is advantageous with respect to solution dansylation. It improves the total analysis time, avoids high temperatures that can affect the thermal stability of the derivatives and could make possible the automation of the procedure.

Peptide Metal–Organic Frameworks for Enantioselective Separation of Chiral Drugs

We report the use of a chiral Cu(II) 3D metal-organic framework (MOF) based on the tripeptide Gly-l-His-Gly (GHG) for the enantioselective separation of metamphetamine and ephedrine. Monte Carlo simulations suggest that chiral recognition is linked to preferential binding of one of the enantiomers as a result of either stronger or additional H-bonds with the framework that lead to energetically more stable diastereomeric adducts. Solid-phase extraction of a racemic mixture by using Cu(GHG) as the extractive phase permits isolating >50% of the (+)-ephedrine enantiomer as target compound in only 4 min. To our knowledge, this represents the first example of a MOF capable of separating chiral polar drugs.

Preconcentration of emerging contaminants in environmental water samples by using silica supported Fe3O4 magnetic nanoparticles for improving mass detection in capillary liquid chromatography

A magnetic material based on Fe(3)O(4) magnetic nanoparticles incorporated in a silica matrix by using a sol-gel procedure has been used to extract and preconcentrate emerging contaminants such as acetylsalicylic acid, acetaminophen, diclofenac and ibuprofen from environmental water samples prior to the analysis with Capillary LC-MS. The use of the proposed silica supported Fe(3)O(4) magnetic nanoparticles enables surfactant free extracts for the analysis with MS detection without interferences in the ionisation step. Under the optimum conditions, we demonstrated the reusability of the magnetic sorbent material during 20 uses without loss in the extraction efficiency. In addition, no cleanup was necessary. The preconcentration factor was 100 and the detection limits were between 50 and 150 ng/L. The proposed procedure has been applied to the analysis of water samples obtaining recoveries between 80 and 110% and RSD values lower than 12%. Concentrations of the target analytes over the range 1.7 and 0.1 μg/L have been found in different water samples.

Miniaturized matrix solid phase dispersion procedure and solid phase microextraction for the analysis of organochlorinated pesticides and polybrominated diphenylethers in biota samples by gas chromatography electron capture detection

This work has developed a miniaturized method based on matrix solid phase dispersion (MSPD) using C18 as dispersant and acetonitrile-water as eluting solvent for the analysis of legislated organochlorinated pesticides (OCPs) and polybrominated diphenylethers (PBDEs) in biota samples by GC with electron capture (GC-ECD). The method has compared Florisil-acidic Silica and C18 as dispersant for samples as well as different solvents. Recovery studies showed that the combination of C18-Florisil was better when using low amount of samples (0.1 g) and with low volumes of acetonitrile-water (2.6 mL). The use of SPME for extracting the analytes from the solvent mixture before the injection resulted in detection limits between 0.3 and 7.0 microg kg(-1) (expressed as wet mass). The miniaturized procedure was easier, faster, less time consuming than the conventional procedure and reduces the amounts of sample, dispersant and solvent volume by approximately 10 times. The proposed procedure was applied to analyse several biota samples from different parts of the Comunidad Valenciana.

Generalized H-point standard additions method for analyte determinations in unknown samples

The generalized H-point standard additions method (GHPSAM) is proposed. The method locates the linear spectral behaviour for the unknown global interference, if it exists, in all the spectral regions measured. The linear spectral interval for the interference is located from second derivative data. After this, the method validates the linearity supposition and estimates the analyte concentration free from bias error. So, only the data measured from the method are enough to obtain the unbiased analyte concentration in the presence of unknown interferences. Mixtures of phenol and 4-chloro-2-nitrophenol, 2,4-dichlorophenol and 2,4,6-trichlorophenol have been tested in order to check the validity of the method. The method also has been applied to the determination of amphetamine in urine samples. It works well in a lot of spectral situations, except in the cases when the global interference and the analyte are linear in the same spectral region.