Characterisation and determination of fullerenes: a critical review

A prominent sector of nanotechnology is occupied by a class of carbon-based nanoparticles known as fullerenes. Fullerene particle size and shape impact in how easily these particles are transported into and throughout the environment and living tissues. Currently, there is a lack of adequate methodology for their size and shape characterisation, identification and quantitative detection in environmental and biological samples. The most commonly used methods for their size measurements (aggregation, size distribution, shape, etc.), the effect of sampling and sample treatment on these characteristics and the analytical methods proposed for their determination in complex matrices are discussed in this review. For the characterisation and analysis of fullerenes in real samples, different analytical techniques including microscopy, spectroscopy, flow field-flow fractionation, electrophoresis, light scattering, liquid chromatography and mass spectrometry have been reported. The existing limitations and knowledge gaps in the use of these techniques are discussed and the necessity to hyphenate complementary ones for the accurate characterisation, identification and quantitation of these nanoparticles is highlighted

Non-aqueous capillary electrophoresis separation of fullerenes and C60 fullerene derivatives

As the interest in the use of fullerene compounds in biomedical and cosmetic applications increases, so too does the need to develop methods for their determination and quantitation in such complex matrices. In this work we studied the behavior of C60 and C70 fullerenes in non-aqueous capillary electrophoresis, as well as two C60 fullerene derivatives not previously reported by any electrophoretic method, N-methyl-fulleropyrrolidine (C60 pyrr) and (1,2-methanofullerene C60)-61-carboxylic acid (C60-COOH). . The separation was performed using fused-silica capillaries with an I.D. of 50 µm and tetraalkylammonium salts, namely tetra-n-decylammonium bromide (TDAB, 200 mM) and tetraethylammonium bromide (TEAB, 40 mM), in a solvent mixture containing 6% methanol and 10% acetic acid in acetonitrile:chlorobenzene (1:1 v/v) as the background electrolyte (BGE). Detection limits, based on a signal-to-noise ratio of 3:1, were calculated and values between 1 and 3.7 mg/L were obtained. Good run-to-run and day-to-day precisions on concentration were achieved with RDSs lower than 15%. For the first time, an electrophoretic technique (NACE) has been applied for the analysis of C60 fullerene in a commercial cosmetic cream. A standard addition method was used for quantitation and the result was compared with that obtained by analyzing the same cream by LC-MS

Solid-phase extraction using molecularly imprinted polymer for selective extraction of natural and syntetic estrogens from aqueous samples

A method is proposed for the clean-up and preconcentration of natural and synthetic estrogens from aqueous samples employing molecularly imprinted polymer (MIP) as selective sorbent for solid-phase extraction (SPE). The selectivity of the MIP was checked toward several selected natural and synthetic estrogens such as estrone (E1), 17β-estradiol (β-E2), 17α-estradiol (α-E2), estriol (E3), 17α-ethinylestradiol (EE2), dienestrol (DIES) and diethylstilbestrol (DES). Ultrahigh pressure liquid chromatography (UHPLC) coupled to a TSQ triple quadrupole mass spectrometry (QqQ) was used for analysis of target analytes. The chromatographic separation of the selected compounds was performed in less than 2 min under isocratic conditions. The method was applied to the analysis of estrogens in spiked river and tap water samples. High recoveries (>82%) for estrone, 17β-estradiol, 17α-estradiol, estriol and 17α-ethinylestradiol were obtained. Lower but still satisfactory recoveries (>48%) were achieved for dienestrol and diethylstilbestrol. The method was validated and found to be linear in the range 50-500 ng L-1 with correlation coefficients (R2) greater than 0.995 and repeatability relative standard deviation (RSD) below 8% in all cases. For analysis of 100-ml sample, the method detection limits (LOD) ranged from 4.5 to 9.8 ng L-1 and the limit of quantitation (LOQ) from 14.9 to 32.6 ng L-1. To demonstrate the potential of the MIP obtained, a comparison with commercially available C18 SPE was performed. Molecularly imprinted SPE showed higher recoveries than commercially available C18 SPE for most of the compounds. These results showed the suitability of the MIP-SPE method for the selective extraction of a class of structurally related compounds such as natural and synthetic estrogens

In-line preconcentration capillary zone electrophoresis for the analysis of haloacetic acids in water

Two in-line enrichment procedures (large volume sample stacking (LVSS) and field amplified sample injection (FASI)) have been evaluated for the capillary zone electrophoresis (CZE) analysis of haloacetic acids (HAAs) in drinking water. For LVSS, separation on normal polarity by using 20 mM acetic acid-ammonium acetate (pH 5.5) containing 20% acetonitrile as BGE was required. For FASI, the optimum conditions were 25 s hydrodynamic injection (3.5 kPa) of a water plug followed by 25 s electrokinetic injection (-10 kV) of the sample, and 200 mM formic acid-ammonium formate buffer at pH 3.0 as BGE. For both FASI and LVSS methods, linear calibration curves (r2>0.992), limit of detection (LOD) on standards prepared in Milli-Q water (49.1-200 µg/L for LVSS and 4.2-48 µg/L for FASI), and both run-to-run and day-to-day precisions (RSD values up to 15.8% for concentration) were established. Due to the higher sensitive enhancement (up to 310-fold) achieved with FASI-CZE this method was selected for the analysis of HAAs in drinking water. However, for an optimal FASI application sample salinity was removed by solid phase extraction (SPE) using Oasis WAX cartridges. With SPE-FASI-CZE, method detection limits in the range 0.05-0.8 µg/L were obtained, with recoveries, in general, higher than 90% (around 65% for monochloroacetic and monobromoacetic acids). The applicability of the SPE-FASI-CZE method was evaluated by analyzing a drinking tap water from Barcelona where seven HAAs were found at concentration levels between 3-13 µg/L

Analysis of UV ink photoinitiators in packaged food by fast liquid chromatography at subambient temperature coupled to tandem mass spectrometry

A fast method of liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) was developed for the analysis of eleven UV ink photoinitiators in packaged food. Chromatographic separation was achieved in a pentafluorophenylpropyl (PFPP) column at 5ºC and acetonitrile:25 mM formic acid-ammonium formate (pH 2.7) in gradient elution. To reduce sample treatment, a QuEChERS (quick, easy, cheap, effective, rugged and safe) method for the extraction and clean-up of UV photoinitiators in packaged foods was evaluated. Triple quadrupole working in H-SRM on Q1 mode was used for both quantitation and confirmation purposes and the most intense and selective transitions were chosen. Quality parameters of the developed QuEChERS LC-MS/MS method were established and applied for the analysis of photoinitiators in food packaged at ng kg-1 levels

5-Hydroxymethylfurfural content in foodstuffs determined by micellar electrokinetic chromatography

Micellar electrokinetic chromatography (MEKC) has been applied for the determination of 5-hydroxymethylfurfural in several foodstuffs. A 75 mM phosphate buffer solution at pH 8.0 containing 100 mM sodium dodecylsulfate was used as background electrolyte (BGE), and the separation was performed by applying +25 kV in a 50 µm I.D. uncoated fused-silica capillary. Good linearity over the range 2.5-250 mg kg-1 (r2 ≥ 0.999) and run-to-run and day-to-day precisions at low and medium concentration levels were obtained. Sample limit of detection (0.7 mg kg-1) and limit of quantification (2.5 mg kg-1) were established by preparing the standards in blank matrix. The procedure was validated by comparing the results with those obtained with liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Levels of HMF in 45 different foodstuffs such as breakfast cereals, toasts, honey, orange juice, apple juice, jam, coffee, chocolate and biscuits were determined

Strategies for the multi-residue analysis of 100 pesticides by liquid chromatography-triple quadrupole mass spectrometry

Analytical strategies for screening, quantitation and confirmation of a group of 100 pesticides in fruit and vegetable samples by LC-MS and LC-MS/MS were developed. The pesticides studied belong to different chemical families of herbicides, insecticides and fungicides. A selection of some degradation products was also included. Chromatographic separation was performed using a Zorbax Eclipse XDB-C8 column (150 mm × 4.6 mm and 5 µm particle size), and gradient elution with acetonitrile-water (both with 0.1% formic acid) as mobile phase. LC-MS/MS using highly-selective selected reaction monitoring (H-SRM) acquisition mode monitoring two transitions for each compound showed to be the most sensitive methodology. Quantitation was carried out using matrix-matched standard calibration and good linearity of response was demonstrated (r > 0.998). Limits of detection (by acquiring two transitions and with the ion-ratio requirements) ranged between 0.01 and 20 µg/kg were obtained. So, in general, the sensitivity achieved meets the maximum residue levels (MRLs) established by the European Union regulation for food monitoring programs. Pesticide confirmation was carried out following European Union guidelines. In order to prevent false-positives, further confirmatory strategies were proposed. LC-MS in highly-selective selected ion monitoring (H-SIM) mode with accurate mass measurement was used to obtain an orthogonal criterion (exact mass) for confirmation. Accurate mass measurements were always bellow 0.9 mDa for almost all pesticides studied (similar to those described with TOF instruments). A user reversed energy ramp (RER) product ion scan spectra library was generated by means of a data dependent analysis for routine library searching of pesticides. The combination of LC-MS/MS in H-SRM mode and the generation of the RER product ion scan spectra and library search were then used to achieve further confirmation on pesticide analysis. The LC-MS and LC-MS/MS strategies developed were successfully applied for the analysis and confirmation of pesticides in different types of fruit and vegetables samples, and examples of the screening, quantitation and confirmation of pesticides in these samples are shown in this work

Chloride-attachment atmospheric pressure photoionisation for the determination of short-chain chlorinated paraffins by gas chromatography-high-resolution mass spectrometry

In this work, a new gas chromatography-high-resolution mass spectrometry (GC-HRMS) method based on atmospheric pressure photoionisation (APPI) has been developed for the accurate determination of short-chain chlorinated paraffins (SCCPs) as a reliable alternative to the established methods. To the best of our knowledge, this is the first time these compounds has been analysed by GC-MS using atmospheric pressure photoionisation (APPI). Efficient ionisation of SCCPs was achieved using the new GC-APPI source by the formation of [MþCl]e adduct ions in negative ion mode using dopant-assisted APPI with a mixture of acetone/CCl4 (3:1, v/v). Operating at a resolution of 70,000 FWHM (full width at half maximum) and monitoring the [MþCl]e adduct ions for each congener group, a selective determination of the SCCPs was achieved, avoiding isobaric interferences between homologue groups with different carbon chain length and chlorination degree. Moreover, the GC-APPI-HRMS response of each congener group was mainly influenced by its concentration and did not depend on the number of chlorine atoms in the molecule as occurs with the GC-MS methods based on the electron-capture negative ionisation (ECNI). Thus, the contribution of the different carbon and chlorine homologue groups in the SCCP mixtures was determined by the internal normalization method, and the quantification was performed independently of the chlorine content of the SCCP standard mixture employed. The developed GC-APPI-HRMS method offers some interesting advantages over the existing methods, particularly the possibility to quantify individual SCCP congener groups, the use of a simple calibration method for quantification, and an important timesaving in the data processing, especially over ECNI-based traditional methods. The GC-APPI-HRMS method allowed the determination of SCCPs at low concentration levels in fish samples with lowmethod limits of detection (17e34 pg g1 wet weight for total SCCPs), good precision (RSD < 7%) and trueness (relative error < 8%) and can be proposed as a reliable alternative of the established methods for the determination of these pollutants in environmental sample

New branches in the degradation pathway of monochlorocatechols by Aspergillus nidulans: a metabolomics analysis

A collective view of the degradation of monochlorocatechols in fungi is yet to be attained, though these compounds are recognised as key degradation intermediates of numerous chlorinated aromatic hydrocarbons, including monochlorophenols. In the present contribution we have analysed the degradation pathways of monochlorophenols in Aspergillus nidulans using essentially metabolomics. Degradation intermediates herein identified included those commonly reported (e.g. 3-chloro-cis,cis-muconate) but also compounds never reported before in fungi revealing for 4-chlorocatechol and for 3- chlorocatechol unknown degradation paths yielding 3-chlorodienelactone and catechol, respectively. A different 3-chlorocatechol degradation path led to accumulation of 2- chloromuconates (a potential dead-end), notwithstanding preliminary evidence of chloromuconolactones and protoanemonin simultaneous formation. In addition, some transformation intermediates, of which sulfate conjugates of monochlorophenols/ chlorocatechols were the most common, were also identified. This study provides critical information for understanding the role of fungi in the degradation of chlorinated aromatic hydrocarbons; furthering their utility in the development of innovative bioremediation strategies

Preventing false negatives with high-resolution mass spectrometry: the benzophenone case

Benzophenone (BP) is one of the many contaminants reported as present in foodstuff due to its migration from food packaging materials. Liquid chromatography tandem mass spectrometry (LC-MS/MS) is acknowledged in the literature as the method of choice for this analysis. However, cases have been reported where the use of this methodology was not enough to unambiguously confirm the presence of a contaminant. In previous work performed by the authors, the unequivocal identification of BP in packaged foods was not possible even when monitoring two m/z transitions, since ion ratio errors higher than 20% were obtained. In order to overcome this analytical problem a fast, sensitive and selective liquid chromatography-high resolution-mass spectrometry (LC-HRMS) methodology has been developed and applied to the analysis of BP in packaged foods. A direct comparison between liquid chromatography high resolution mass spectrometry (LC-HRMS) and LC-MS/MS data indicated better selectivity when working with LC-HRMS at a resolving power of 50,000 FWHM than when monitoring two m/z transitions by LC-MS/MS. The resolving power used enabled the detection and identification of Harman as the compound impeding the confirmation of BP by LC-MS/MS. Similar quantitative results were obtained by an Orbitrap mass analyser (Exactive ¿) and a triple quadrupole mass analyser (TSQ Quantum Ultra AM ¿)