New Analytical Methodologies based on Chromatography-Atmospheric Pressure Ionization-Mass Spectrometry for the Determination of Halogenated Organic Contaminants

Abstract

[eng] The environment sustainability is being threatened by the continuous release of pollutants that can negatively affect not only environmental compartments but also wildlife and human beings. Among these pollutants, halogenated persistent organic pollutants and new emerging contaminants have cause great concern due to their toxicity, persistence, bioaccumulation and biomagnification capacity and/or their high mobility in the environment. Moreover, their hazardous effects are manifested even at trace levels, thus requiring very selective and sensitive analytical methodologies to face their detection in environmental samples. In this sense, atmospheric pressure ionization (API) sources such as atmospheric pressure chemical ionization (APCI) and atmospheric pressure photoionization (APPI) for both liquid-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS) could offer great advantages to overcome the limitations observed in the determination of these group of substances. In the present Thesis, the feasibility of these ionization sources, especially APPI, has been evaluated to develop sensitive and selective LC-API-MS and GC-API-MS methodologies to monitor relevant halogenated contaminants in environmental samples. API sources have been thoroughly tested to achieve an efficient ionization of neutral per- and polyfluoroalkyl substances (nPFAS). The ionization behavior of these compounds was assessed through optimization of the mobile phase composition, the addition of additives or dopants as well as critical ion source working parameters. These studies have led to highly selective and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) and gas chromatography-high- resolution mass spectrometry (GC-HRMS) methodologies (up to fg injected on column). Furthermore, an efficient solid-phase extraction and a fast and in-situ preconcentration headspace solid-phase microextraction procedures have been developed to analyze nPFAS in river water samples avoiding analyte losses observed during evaporation steps. Additionally, the fragmentation pathways of ions generated for these compounds in API sources has been tentatively proposed using the combined information of in-source fragmentation, tandem mass spectrometry and high-resolution mass spectrometry. These fragmentation pathways aim to provide useful information for the development of target, suspect and non-targeted analysis strategies for the identification of known and new families of nPFAS in complex environmental samples. Furthermore, in this Thesis, the novel GC-APPI-HRMS (Orbitrap) system is proposed to face the main limitations that have been observed in the currently used analytical determinations of relevant chlorinated contaminants such as dechlorane plus (DP) and analogs, polychlorinated naphthalenes (PCNs), polychlorinated dibenzo-ρ-dioxins and dibenzofurans (PCDD/Fs), dioxin-like polychlorinated biphenyls (dl- PCBs), and short-chain chlorinated paraffins (SCCPs). The soft ionization of GC-APPI has been used to promote molecular or quasi-molecular ions as well as characteristic cluster ions such as [M‒Cl+O]‒, allowing the development of sensitive and selective methods. The use of dopant vapors has been thoroughly investigated to detect the critical parameters that allow maximizing the ionization efficiency of the analytes. Additionally, anion-attachment ionization strategies have been also studied to reduce the in-source fragmentation and to improve sensitivity and selectivity for SCCPs. Furthermore, multidimensional separation strategies (using novel stationary phases and/or ion mobility separation) have been also evaluated to improve the separation of those compounds that often coelute (PCNs or SCCPs).The GC- APPI-HRMS methods developed in this Thesis have shown a great detection capability (up to low fg injected on column) and a high selectivity due to both the exact mass measurements (Orbitrap) and the soft ionization provided by the GC-APPI source. Moreover, they have demonstrated a good performance to determine these compounds in marine sediments, fly ashes, gull eggs, or fishes among other complex environmental samples.

[spa] La incesante emisión de contaminantes supone una amenaza tanto para el medio ambiente como para los seres vivos. Entre los contaminantes que han despertado un mayor interés ambiental destacan los compuestos orgánicos halogenados debido a su alta toxicidad a bajas concentraciones, persistencia, capacidad de bioacumulación y para ser transportados a largas distancias. La monitorización de estos contaminantes a bajos niveles de concentración en muestras ambientales requiere de metodologías analíticas selectivas y con una alta capacidad de detección. Así, en esta Tesis se ha evaluado la capacidad de las fuentes de ionización química (APCI) y la fotoionización a presión atmosférica (APPI) para ionizar eficientemente estas familias de contaminantes y así proponer métodos basados en cromatografía de líquidos y de gases acopladas a la espectrometría de masas en tándem y/o de alta resolución (LC-MS/MS y GC-HRMS). Los estudios desarrollados durante esta Tesis han permitido establecer condiciones de trabajo que reducen la fragmentación en la fuente y aumentan la respuesta de los iones generados. Para ello, se han evaluado la composición de la fase móvil, la adición de aditivos que influyen en la ionización de los analitos y diversas condiciones de trabajo, permitiendo establecer tendencias en la ionización de estos contaminantes, haciendo especial énfasis en la novedosa fuente de ionización del sistema GC-APPI-HRMS. Además, se ha estudiado la fragmentación MS/MS de estos compuestos, establecido rutas de fragmentación e identificado/caracterizado los iones observados. Estos estudios han permitido desarrollar métodos que ofrecen una solución a algunas de las limitaciones observadas en las metodologías existentes, mejorando la selectividad y la capacidad de detección. Además, los estudios de ionización y el uso de la MS/MS y de la HRMS han permitido desarrollar estrategias de análisis no dirigido para facilitar la identificación en muestras complejas de compuestos similares a los estudiados en esta Tesis.