Nuria Cortés-Francisco

Targeted analysis with benchtop quadrupole-orbitrap hybrid mass spectrometer: application to determination of synthetic hormones in animal urine.

Sensitive and unequivocal determination of analytes/contaminants in complex matrices is a challenge in the field of food safety control. In this study, various acquisition modes (Full MS/AIF, Full MS+tMS/MS, Full MS/dd MS/MS and tSIM/ddMS/MS) and parameters of a quadrupole-orbitrap hybrid mass spectrometer (Q Exactive) were studied in detail. One of the main conclusions has been that, reducing the scan range for Full MS (using the quadrupole) and targeted modes give higher signal-to-noise (S/N) ratios and thereby better detection limits for analytes in matrix. The use of Q Exactive in a complex case, for the confirmatory analysis of hormones in animal urine is presented. A targeted SIM data dependent MS/MS (tSIM/ddMS/MS) acquisition method for determination of eight synthetic hormones (trenbolone, 17α ethinylestradiol, zeranol, stanozolol, dienestrol, diethylstilbestrol, hexestrol, taleranol) and a naturally occurring hormone (zearalenone) in animal urine were optimized to have sensitive precursors from targeted SIM mode and trigger MS/MS scans over the entire chromatograph peak. The method was validated according to EC/657/2002. CCα (decision limit) for the analytes ranged between 0.11 μg L(-1) and 0.69 μg L(-1) and CCβ (detection capability) ranged between 0.29 μg L(-1) and 0.90 μg L(-1).

Liquid chromatography coupled to tandem mass spectrometry and high resolution mass spectrometry as analytical tools to characterize multi-class cytostatic compounds

Cytostatic compounds used in the treatment of cancer have emerged as a new generation of water contaminants due to the continuous amounts administered to patients and to the fact that a variable proportion is excreted unchanged. In this study, we have evaluated the performance of liquid-chromatography-tandem mass spectrometry (LC-MS/MS) and high resolution mass spectrometry using an Orbitrap analyzer (LC-HRMS) for the multiresidue determination of multi-class cytostatic compounds. In a first step, ionization conditions were tested in positive electrospray mode and optimum fragmentation patterns were determined. For LC-MS/MS, two selected reaction monitoring (SRM) transitions were optimized and for LC-HRMS, the molecular ion with 5 ppm error and two product ions were defined. Following, the chromatographic conditions were optimized considering that compounds analyzed have a very different chemical structure and chromatographic behavior. The best performance was obtained with a Luna C18 column, which permitted the separation of the 26 compounds in 15 min. Finally, the performance of LC-MS/MS and LC-HRMS was compared in terms of linearity, sensitivity, intra and inter-day precision and overall robustness. While LC-MS/MS provided good identification capabilities due to selective SRM transitions, LC-Orbitrap proved to be 100 times more sensitive. This study provides a comprehensive overview on the MS conditions to determine the outmost used cytostatic compounds and provides a spectral library to be used for the identification of these compounds in water or biological matrices.

Determination of lipophilic marine toxins in mussels. Quantification and confirmation criteria using high resolution mass spectrometry

A multitoxin method has been developed for quantification and confirmation of lipophilic marine biotoxins in mussels by liquid chromatography coupled to high resolution mass spectrometry (HRMS), using an Orbitrap-Exactive HCD mass spectrometer. Okadaic acid (OA), yessotoxin, azaspiracid-1, gymnodimine, 13-desmethyl spirolide C, pectenotoxin-2 and Brevetoxin B were analyzed as representative compounds of each lipophilic toxin group. HRMS identification and confirmation criteria were established. Fragment and isotope ions and ion ratios were studied and evaluated for confirmation purpose. In depth characterization of full scan and fragmentation spectrum of the main toxins were carried out. Accuracy (trueness and precision), linearity, calibration curve check, limit of quantification (LOQ) and specificity were the parameters established for the method validation. The validation was performed at 0.5 times the current European Union permitted levels. The method performed very well for the parameters investigated. The trueness, expressed as recovery, ranged from 80% to 94%, the precision, expressed as intralaboratory reproducibility, ranged from 5% to 22% and the LOQs range from 0.9 to 4.8pg on column. Uncertainty of the method was also estimated for OA, using a certified reference material. A top-down approach considering two main contributions: those arising from the trueness studies and those coming from the precisions determination, was used. An overall expanded uncertainty of 38% was obtained.

Molecular Characterization of Dissolved Organic Matter through a Desalination Process by High Resolution Mass Spectrometry

The effect of different water treatments such as ultrafiltration (UF) and reverse osmosis (RO) on dissolved organic matter (DOM) is still unknown. Electrospray ionization Fourier transform orbitrap mass spectrometry has been used to provide valuable information of marine DOM evolution through a desalination process on a molecular scale. In the present manuscript, the characterization of four real composite water samples from a desalination pilot plant installed in the coast of Barcelona (Spain) has been carried out. The sampling was performed on each point of the pilot plant: raw seawater (RSW), UF effluent, brine RO and permeate RO. The mass spectra of the different samples show several thousand peaks, however for the present screening study, only the mass range m/z 200-500 and the main signals in this mass range (relative intensities ≥1%) have been considered. The analysis of RSW and UF samples reveal that there is little effect on DOM by the UF pilot. However, when the water is treated on the RO an important change on DOM has been observed. The recurring periodical patterns found in RSW and UF are lost in Permeate RO sample. Compounds with more aliphatic character, with higher H/C ratio (H/Cav 1.72) are present in the Permeate and some of them have been tentatively identified as fatty acids.

Comparison of triple quadrupole mass spectrometry and Orbitrap high-resolution mass spectrometry in ultrahigh performance liquid chromatography for the determination of veterinary drugs in sewage: benefits and drawbacks

This paper presents a comparison of triple quadrupole tandem mass spectrometry (MS/MS) and Orbitrap high-resolution mass spectrometry (HRMS) combined to ultrahigh performance liquid chromatography for the determination of glucocorticoids and polyether ionophores in sewage, in order to show the major benefits and drawbacks for each mass spectrometry analyser. Overall, HRMS measurements have enhanced performance in terms of confirmatory capabilities than MS/MS measurements. Moreover, similar limits of quantification, limits of detection, linear range and repeatability for glucocorticoids with both the MS/MS and HRMS methods were compared, but in the case of polyether ionophores, slightly better limits of detection and limits of quantification were obtained with the HRMS method because of the high sensitivity obtained when diagnostic ions are used for quantification instead of selected reaction monitoring transitions for these compounds. The two methods have been applied to the analysis of several influent and effluent sewage samples from sewage treatment plants located in the Tarragona region (Catalonia, Spain), showing an excellent correlation between the two methods.

New method for the analysis of lipophilic marine biotoxins in fresh and canned bivalves by liquid chromatography coupled to high resolution mass spectrometry: a quick, easy, cheap, efficient, rugged, safe approach.

A new method for the analysis of lipophilic marine biotoxins (okadaic acid, dinophysistoxins, azaspiracids, pectenotoxins, yessotoxins, spirolids) in fresh and canned bivalves has been developed. A QuEChERS methodology is applied; i.e. the analytes are extracted with acetonitrile and clean-up of the extracts is performed by dispersive solid phase extraction with C18. The extracts are analyzed by ultra-high performance liquid chromatography coupled to a hybrid quadrupole-Orbitrap mass spectrometer, operating in tandem mass spectrometry mode, with resolution set at 70,000 (m/z 200, FWHM). Separation of the analytes, which takes about 10min, is carried out in gradient elution mode with a BEH C18 column and mobile phases based on 6.7mM ammonia aqueous solution and acetonitrile mixtures. For each analyte the molecular ion and 1 or 2 product ions are acquired, with a mass accuracy better than 5ppm. The quantification is performed using surrogate matrix matched standards, with eprinomectin as internal standard. The high-throughput method, which has been successfully validated, fulfills the requirements of European Union legislation, and has been implemented as a routine method in a public health laboratory.

High-field FT-ICR mass spectrometry and NMR spectroscopy to characterize DOM removal through a nanofiltration pilot plant

Ultrahigh resolution Fourier transform ion cyclotron mass spectrometry and nuclear magnetic resonance spectroscopy were combined to evaluate the molecular changes of dissolved organic matter (DOM) through an ultrafiltration-nanofiltration (UF-NF) pilot plant, using two dissimilar NF membranes tested in parallel. The sampling was performed on seven key locations within the pilot plant: pretreated water, UF effluent, UF effluent after addition of reagents, permeate NF 1, permeate NF 2, brine NF 1 and brine NF 2, during two sampling campaigns. The study showed that there is no significant change in the nature of DOM at molecular level, when the water was treated with UF and/or with the addition of sodium metabisulfite and antiscaling agents. However, enormous decrease of DOM concentration was observed when the water was treated on the NF membranes. The NF process preferentially removed compounds with higher oxygen and nitrogen content (more hydrophilic compounds), whereas molecules with longer pure aliphatic chains and less content of oxygen were the ones capable of passing through the membranes. Moreover, slight molecular selectivity between the two NF membranes was also observed.

Ultrahigh resolution mass spectrometry and accurate mass measurements for high‐throughput food lipids profiling

In the present study, accurate mass measurements by ultrahigh resolution mass spectrometry with Orbitrap-Exactive working at resolving power R: 100,000 (m/z 200, full width at half maximum) with an accuracy better than 2 ppm in all the mass range (m/z 200 to 2000) were used to show a detailed molecular composition of diverse edible oils and fats. Flow injection was used to introduce samples into the mass spectrometer, obtaining a complete analysis of each sample in less than 10 min, including blanks. Meticulous choice of organic solvents and optimization of the ion source and Orbitrap mass analyzer parameters were carried out, in order to achieve reproducible mass spectra giving reliable elemental compositions of the lipid samples and to prevent carry over. More than 200 elemental compositions attributable to diacylglycerols, triacylglycerols (TAGs), and their oxidation products have been found in the spectra of food lipids from different origin. Several compounds with very close molecular mass could only be resolved through ultrahigh resolution, allowing detailed and robust TAG profiling with a high characterization potential.

Insight into virgin olive oil secoiridoids characterization by high-resolution mass spectrometry and accurate mass measurements.

With the aim to enhance characterization of virgin olive oil (VOO), high resolution mass spectrometry (HRMS) and high resolution tandem mass spectrometry (HRMS/MS), in positive and negative electrospray ionization (ESI) modes, coupled to fused-core reverse phase chromatography, were applied to distinct VOO phenolic extracts after the optimization of chromatographic conditions, ESI and fragmentation parameters. HRMS, but also HRMS/MS resulted fundamental to progress in secoiridoids structural elucidation. The former revealed that the secoiridoid composition of VOO was far more complicated than previously reported, while the latter helped clarify product ion elemental composition allowing new fragmentations, in addition to those reported in the literature, to be put forward. In particular, for the first time, different product ions with the same nominal mass were unequivocally identified in the spectra of secoiridoid compounds, confirming the greater capacity of HRMS/MS to clarify structure than low-resolution MS. Furthermore, and differing from previous studies, the multiple isomers of the main VOO secoiridoids could be differentiated on the basis of their HR product ion spectra in positive mode.

Fragmentation studies for the structural characterization of marine dissolved organic matter

AbstractHigh-resolution tandem mass spectrometry by collision-induced dissociation with a linear ion trap-Orbitrap has been performed on marine dissolved organic matter (DOM). Product ion spectra of selected precursor ions (m/z 359–375) have been acquired to obtain structural information, after method development. To evaluate the performance of the method, the Suwannee River fulvic acid (SRFA) reference standard was also analyzed. By reconstructing the individual product ion spectrum of marine DOM, several fragments were assigned to the different precursor ions indicating the presence of carboxyl, hydroxyl, lactones, quinones, esters, and structures more similar to lignin-degraded molecules. On the basis of these findings, coastal marine DOM molecules, although structurally homogeneous, might be more rich in diversity of functional groups than previously described. Graphical abstractReconstructed product ion spectrum to assemble the DOM structure