Extraction system and determination of PAH in seawater and it implementation in submarine vehicles

Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. Luis Aarón García Ortega Curso 2015/2016 Tutor: Daura Vega Moreno Trabajo Fin de Título para la obtención del título de Graduado en Ciencias del Mar. Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. -2 - Sistemas de extracción y determinación de hidrocarburos en agua de mar y su acoplamiento en vehículos submarinos Datos del estudiante Nombre: Luis Aarón Apellidos: García Ortega Titulación: Grado en Ciencias del mar. Datos del tutor, Departamento y Empresa Tutor: Daura Vega Moreno Departamento: Departamento de Química Empresa: Universidad de Las Palmas de Gran Canaria (ULPGC) Firma del estudiante Firma del tutor Fecha: 25.05.2016 Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 3 - Table of contents 1. Introduction 5 2. Material and methods 9 2.1 Elaboration of the standards 9 2.2 Preparation of the samples 9 2.3 Instrumentation 9 2.4 Chromatographic Analysis 10 2.5 Preparation of extraction system and it optimization 11 2.6 Summary of the procedure of extraction and determination 12 3. Results 13 3.1 Preselection of solid sorbent (SPE) 13 3.2 Optimization of the extraction system 15 3.3 Limit of detection and reproducibility 16 3.4 Applications 17 4. Discussion 19 5. Conclusion 20 6. Bibliography 20 Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 4 - Abstract Policyclyc aromatic hydrocarbons (PAHs) are a potential risk for human health and marine biota in general that make necessary the monitorization of them. A miniaturized extraction system capable to extract PAHs from seawater was developed and optimized with the objective of implement it in an oceanographic buoy in the future. An analytical method was optimized by high performance liquid chromatography (HPLC) for the determination of extracted PAHs by the extraction system. The analytical method was validated and applicated to real samples of differents points of Gran Canaria. The method has enough sensitivity to detect and quantify concentrations below the concentrations established in the legislation. In some places where samples were taken some compounds exceed the legislation while other compounds follow it. The results obtained in this work and included in this TFT will be sent for it publication in the “Analytica Chimica Acta” magazine, the first quartil in the area of analytical chemistry. Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 5 - 1. Introduction Polycyclic Aromatic Hydrocarbons (PAH) are compounds formed by two or more aromatic rings in different arrangements [1]. They are considered ubiquitous compounds due to their persistence in the environment and they have several degrees of toxicity and persistence [2]. It has demonstrated that some of these compounds have carcinogenic, mutagenic and/ or teratogenic properties and constitute a potential risk for human health [3][4]. For this reason it is very important to control and monitorize them. To facilitate this labor the United States Environmental Protection Agency (USEPA) has elaborated a list in which 16 PAHs of most interest due to their toxicity and dangerous were listed. The European Commission has regulated the monitorization of these compounds through the Water framework directive (2000/60/CE). This directive has been modified and this modification can be found in the directive (2008/105/CE) which regulates the environment quality norms for dangerous priority substances and other contaminants [5]. In this directive are listed 5 PAHs as dangerous priority substances. These are benzo[a]pyrene, benzo[b]fluorenthene, benzo[g,h,i]perylene, benzo[k]fluoranthene, indene[1,2,3cd]pyrene. The two main properties of PAHs are their hydrophobic character and their low vapor pressure. These two properties vary with the molecular weight of the compound being more apolar compounds of high molecular weight and more volatile compounds of low molecular weight [1]. These properties control the presence and distribution of PAHs in the marine environment. When hydrophobic character increase the trend of compounds to adsorb onto particles also increase [3]. Each compound presents a characteristic absorption spectrum. This property facilitates the identification of each compound. The wavelength of the absorption increases with the number of aromatic rings of the compound [6]. Many PAHs have fluorescence and this permit determine them with more sensitivity and selectivity [2]. They can be produced and released to the environment by natural or anthropogenic sources. PAHs present several origins, although the two main are petrogenic and pyrogenic origin. Pyrogenic PAHs present high molecular weight than petrogenic PAHs [7][8]. Several ratios are been used in function of analyzed compounds to determine their origin [3][9][10]. The 16 PAHs of the USEPA can be classified as low molecular weight PAHs (2 to 3 rings) that include naphtalene, acenaphtylene, fluorene, acenaphtene, phenantrene and anthracene. As high molecular weight PAHs (4 to 6 rings) the classify includes fluoranthene, pyrene, benzo[a]anthracene, chrysene, benzo[b] fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, indene[1,2,3-c,d]pyrene, benzo[g,h,i]perylene and dibenz[a,h]anthracene. Some of compounds used in this study are included in the figure 1. Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 6 - Figure 1: Some PAHs included in the USEPA list and used in this study. In the marine environment the main inputs of PAHs are atmospheric, rivers, ports and waste waters ways [2][10]. The concentration of these compounds is higher in coast water than in the open ocean. The distribution of PAHs depends to the properties cited previously [3]. A positive correlation between the different PAHs of high molecular weight and the amount of total organic carbon (TOC) in sediment samples has been observed [11]. This fact suggests an enrichment of these compounds in the sediment zone and due to the dynamic of these contaminants this zone can become a contaminant reservoir. However, low molecular weight PAHs are more abundant in superficial waters due to inputs by different ways, especially in the superficial microlayer where the concentration of PAHs are higher than the waters below [12]. The half-life period of low molecular weight PAHs is reduced in the water column due to the degradation. This cause the limitation of low molecular weight PAHs transport to deep waters [3]. Process and fates of some PAHs are shown in the figure 2. The accumulation of these contaminants is facilitated due to their hydrophobic character. For this reason this property is related with the bioacummulation, bioconcentration and biomagnification factors in organisms. High molecular weight PAHs are considered genotoxic due to they can be bioactivated after the biotransformation and generate electrophilic derivate which can react with the DNA chain and produce damage in it [1][13]. The adsorption of these compounds to Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 7 - multiple surfaces increase the bioavailable in the marine environment and producing the introduction of PAHs in the food chain [9][11]. Figure 2: Process and fate of petrogenic PAHs in this case. Extracted and modified from [14]. In the marine environment the main exposure route is through the gills or the aliment. The concentration of contaminants in organism depends to the metabolism of it [1][11]. Invertebrates lack of an efficient mechanism to remove PAHs from their organism producing bioaccumulation into them. Some of them such as bivalves mollusks are used to monitorize these contaminants in the marine environment in projects such as the Biological effects of environmental pollution in marine coastal systems: European programme (BIOMAR) [9]. Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 8 - The most used analytical techniques for determination of these compounds are the Gas Chromatography (GC) or the High Performance Liquid Cromatography (HPLC) [15]. By HPLC several isomers of different PAHs can be separated and needless a previous clean-up for analysis [16][17]. Different methods of detection can be used with these two techniques. For HPLC the most used are UV-visible and fluorescence detection. The fluorescence method provides higher selectivity and sensitivity than the UV-visible method [6]. An extraction and preconcentration step previous to the analysis is needed due to the very low concentration of PAHs in seawater. In the case of seawater the salinity and the presence of fulvic and humic acid affect the adsorption of these compounds to the particulate phase. For this reason seawater should be prefiltered before the preconcentration step [6]. There are several methods of preconcentration such as liquid-liquid extraction (LLE), solid phase extraction (SPE), supercritical fluid extraction (SFE) and (SPE/SFE). The SPE obtains better results than the others [16]. Currently this technique is usually used due to it higher advantages such as the use of small volumes of organic solvent, diminish the time of the preconcentration and the high available of sorbents in the market that help to the optimization of the process and the reduce of costs [16]. For this study the SPE method has been selected for the extraction and preconcentration step but with a modification and a miniaturization. For the optimization of the method is necessary the study of the conditions of the extraction such as the composition of the fill, the volume of the sample and the amount of solid sorbent used [16]. The extraction and monitorization of these compounds is one of the objectives of this study to medium term and the optimization of the extraction is a critical step for carry out it. Currently, several experiences exist for monitorization of PAHs using optic sensors coupled to AUV but not for take samples in situ for it posterior analysis in laboratories. The Monterey Bay Aquarium Research Institute (MBARI) deployed an AUV to characterize the deep plume of oil spill during the disaster of the Deepwater Horizont to improve the understanding of this phenomenon. As a result a persistent plume was obtained and related with the origin of the spill [18][20]. Recently, fluorescence sensors are beginning to be used, for example in (S. Jocis and F. Pierce) some of them are tested for couple anyone to a Bluefin spray glider [18]. Other systems used are the autosamplers. These systems consist in a silicon rubber surface coated by a semipermeable membrane by which contaminants present affinity. An application of this sampler can be seen in (L. Suberg et al. 2014) [21]. Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 9 - The main objective of this work is the extraction of PAHs from seawater by active systems for it posterior analysis by HPLC. The extraction has been realized by the optimization of a miniaturized SPE system as a basis for future studies that allow the automatization and coupled to a submarine vehicle or an oceanographic buoy. 2. Material and methods 2.1 Elaboration of the standards The compounds used as standards were acenaphthylene (Acen), fluorene (Flren), phenantrene (Phe), anthracene (Anth), fluoranthene (Flran), benzo(a)anthracene7,12-dione (B(a)an), chrysene (Chry) y benzo(b)fluoranthene (B(b)flu) purchased in Sigma-Aldrich® and benzo(a)pyrene (B(a)pyr) from Supelco®. These compounds were diluted in acetonitrile in concentrations among 700 and 1000 ppm attending to their maximum solubility value in the dissolvent. An individual secondary standard of 2ppm was prepared for each compound with methanol. The simultaneous analysis for all compounds was carried out with a tertiary standard of 0.5 ppm in methanol. The reagents used as methanol and acetonitrile were of Panreac® HPLC gradient quality. 2.2 Preparation of samples The seawater that was used for the optimization was collected directly from a water intake in the installations of the Plataforma Oceánica de Canarias (PLOCAN) with a previous filtered step. Seawater was collected every day during the period of the optimization and the validation of the method. Seawater was filtered with a glass fiber filter Whatman® GF/C (porosity of 1.2 μm) before the spike of the sample for the optimization and the extraction method. The spike was realized to 5, 3, 1 and 0.1 ppb of concentration in the seawater. These standards were made with Mili-Q water as solvent. 2.3 Instrumentation The active extraction system includes a peristaltic pump of ISMATEC® trade mark, model: ISM 846, with connectors of SKALAR® trade mark, model: 3091 for each channel of the pump. The miniaturized SPE extraction system was connected to a neoprene tube of 4.8 mm of diameter as is detailed in the section 2.5. Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 16 - Figure 11: Recoveries for the optimization of the weight. 3.3 Limit of detection and reproducibility. The limit of detection was realized two times with artificial seawater. The reproducibility was carried out 7 times with prefiltered seawater and with a spike of a known concentration of 1 ppb. For the reproducibility and the limit of detection the procedure detailed in the section 2.2 was carried out. The concentration of compounds detected in the artificial seawater (control) was calculated for the limit of detection and the limit of quantification. This value was multiplied by 3 for the limit of detection and by 10 for the limit of quantification. Finally, units were converted to ppb (µg/L). The reproducibility was calculated with a mean of tests. Results are shown in the table 2. Compound % RSD LOD (µg/L) LOQ (µg/L) Acen 3,07 2,64 8,81 Flren 5,20 0,07 0,23 Phe 7,25 0,12 0,40 Anth 10,23 0,12 0,42 Flran 3,76 0,02 0,06 B(a)an 11,90 0,06 0,18 Chry 12,84 0,02 0,07 B(b)flu 3,84 0,02 0,05 B(a)pyr 6,06 0,04 0,12 Table 2: The table shows the limit of detection (LOD), limit of quantification (LOQ) and the relative standar deviation (% RSD). 0 20 40 60 80 100 120 140 160 180 Recovery(%) Recoveries per weight 0,1 g 0,2 g 0,3 g Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 17 - The %RSD is below of 10% in most cases with a little differences for chrysene, benzo(b)fluoranthene and anthracene due to their apolar character that make these compounds have low affinity for seawater and tend to adsorb into any surface of the instrumentation. For the limit of detection and the limit of quantification the only compound that presents a limitation in the detection is the acenaphtylene. 3.4 Applications The samples for applications were taken in the places cited in table 2 and shown in figure 12. The seawater was stored in a glass recipient and conserved about 4ºC until it posterior analysis. The analysis was realized two times following the procedure detailed in the section 2.6 but without addition of PAHs through the spike. Site of sample Latitude and Longitude A. Seawater intake of Taliarte (Telde) 27º59’27,49” N 15º22´02,57” O B. Playa de alcaravaneras (Las Palmas de Gran Canaria) 28º07´36,77” N 15º25´31,19” O C. Puerto de las nieves (Agaete) 28º06´01,55” N 15º42´45,89” O D. Muelle deportivo (Las Palmas de Gran Canaria) 28º07´36,77” N 15º25´31,19” O E. Muelle de Taliarte (Telde) 27º59´22,81” N 15º22´07,22” O Table 2: Sites of sample. The results of the applications are shown in the figure 13. In general, low molecular weight PAHs were detected in higher concentrations than the others. High molecular weight PAHs were detected at low concentrations, this is due to their hydrophobic character that makes these compounds adsorb into the particles and accumulate in sediments. As it is shown in figure 13 the Puerto de Las Nieves (Agaete) present the highest concentration for all compounds that were detected except for anthracene. This can be due to the release of these compounds by boats of this port and by maintenance labors of the ferry of Fred Olsen Company. The quiet of these waters facilitate accumulation of these compounds in the port waters. The Muelle Deportivo presents a similar pattern but with lower concentrations than Puerto de Las Nieves (Agaete). For other places, compounds were detected at low concentrations except for anthracene that present a high concentration for all the places. Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 18 - The best concentration for this compound was the seawater intake in the installations of PLOCAN. Figure 12: Localization of samples sites. Figure 13: Aplications of the extraction system. The compounds represented are: 1Fluorene, 2-Phenantrene, 3-Anthracene, 4-Fluoranthene, 5-Benzo(a)anthracene, 6-Chrysene, 7-Benzo(b)fluoranthene. 0,00 1,00 2,00 3,00 4,00 5,00 6,00 7,00 0 1 2 3 4 5 6 7 Concentration (ppb) Compound Aplications Seawater intake Muelle deportivo Playa alcaravaneras Puerto de Las Nieves Muelle Taliarte Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 19 - 4. Discussion A new miniaturized and portable extraction system of contaminants from seawater was developed. It has been validated for the detection of PAH in seawater for different points of Gran Canaria coast. Recoveries are not high (below of 50% in some cases) for the most apolar compounds. This is due to their hydrophobicity and the adsorption of them onto the surface of the instrumentation. Even so a detectable levels below the concentration established in the legislation were observed. For the most polar compounds it can be observed values of recovery up to 120% in some cases. This is due to the used seawater for the optimization that was contaminated with PAHs. Therefore this contamination makes that the known concentration of the added spike was exceed even subtracting the control to obtained recoveries. This is due to a residual concentration below of the limit of detection. For minimize a residual retentions of analytes in the extraction system, the variability in function of the composition of the tubes will be study in the future. In the results a general trend between the recovery and the molecular weight was observed for each compound. High molecular weight PAHs were recoveries at lower concentrations than low molecular weight PAHs. This is due to the hydrophobic character of the first that make these PAHs adsorb onto any surface of the instrumental used in the process. The limit of detection and the limit of quantification are in the range of ppb. These limits are especially low for high molecular PAHs which are a potential risk for human health. Some of these compounds are considered as dangerous priority substances in the legislation so that method is suitable to make analysis in accordance with the concentrations of the directive 2008/105/CE [5]. This guarantees that the method is capable to detect and quantify concentrations below of the concentrations established in the legislation. The directive established the maximum authorized concentrations for anthracene, fluoranthene, benzo(a)pyrene which are 0,4, 1 and 0,1 ppb respectively. For benzo(b)fluoranthene the sum of it concentration and the concentration of it isomer benzo(k)fluoranthene should not exceed 0,03 ppb per year. The values obtained for fluoranthene in the Puerto de Las Nieves (Agate) follow the legislation. However, the concentration of anthracene exceeds the concentration established in the legislation for all the places. The benzo(a)pyrene was not detected in any sample. The benzo(b)fluoranthene was detected only in the Muelle Deportivo and in the intake water of PLOCAN at Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 20 - concentrations of 0,05 and 0,06 ppb. It cannot be determined if this compound follow the legislation due to there is only one data and is necessary the data of a year to calculate the mean of concentrations for this purpose. 5. Conclusion The developed method for extraction of contaminants from seawater has been validated and applicate with successful to determination of PAH in coastal waters of Gran Canaria. This extraction system is miniaturized and portable and will be implemented in oceanographic buoy in the future. The method has been optimized and validated through the determination of PAH by high performance liquid chromatography (HPLC). The method detected and quantified at concentrations below of the concentrations established in the legislation. This method was used for the quantification of PAHs in real samples around of Gran Canaria. With the developed and validated method, it is working in the isolation of the peristaltic pump for couple it into an oceanographic buoy SeaMon-HC of small size and installable in ports. 6. Bibliography [1] M. A. Perugini, M P. Visciano , A. Giammarino , M. Manera , W. Di Nardo , “Polycyclic aromatic hydrocarbons in marine organisms from the Adriatic Sea , Italy,” Chemosphere, vol. 66, pp. 1904–1910, 2007. [2] H. I. Abdel-shafy and M. S. M. Mansour, “REVIEW A review on polycyclic aromatic hydrocarbons : Source , environmental impact , effect on human health and remediation,” Egypt. J. Pet., 2015. [3] T. Chizhova, K. Hayakawa, P. Tishchenko, H. Nakase, and Y. Koudryashova, “Distribution of PAHs in the northwestern part of the Japan Sea,” Deep. Res. Part II Top. Stud. Oceanogr., vol. 86–87, pp. 19–24, 2013. [4] E. M. Basova and V. M. Ivanov, “Modern state of high-performance liquid chromatography of polycyclic aromatic hydrocarbons,” Moscow Univ. Chem. Bull., vol. 66, no. 3, pp. 133–143, 2011. [5] U. Europea, Directiva 2008/105/CE. Relativa a las normas de calidad ambiental en el ámbito de la política de aguas., vol. 348. 2008, pp. 84–97. [6] I. Limam • M. R. Driss, “Off-line solid-phase extraction procedure for the determination of polycyclic aromatic hydrocarbons from aqueous matrices,” Int. J. Environ. Sci. Technol., vol. 10, pp. 973–982, 2013. Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 21 - [7] M. Qiao, C. Wang, S. Huang, D. Wang, and Z. Wang, “Composition, sources, and potential toxicological significance of PAHs in the surface sediments of the Meiliang Bay, Taihu Lake, China,” Environ. Int., vol. 32, no. 1, pp. 28–33, 2006. [8] H. H. Soclo, P. Garrigues, and M. Ewald, “Origin of polycyclic aromatic hydrocarbons (PAHs) in coastal marine sediments: Case studies in Cotonou (Benin) and Aquitaine (France) Areas,” Mar. Pollut. Bull., vol. 40, no. 5, pp. 387–396, 2000. [9] A. Valavanidis, T. Vlachogianni, S. Triantafillaki, M. Dassenakis, F. Androutsos, and M. Scoullos, “Polycyclic aromatic hydrocarbons in surface seawater and in indigenous mussels ( Mytilus galloprovincialis ) from coastal areas of the Saronikos Gulf ( Greece ),” Estuar. Coast. Shelf Sci., vol. 79, pp. 733–739, 2008. [10] M. R. Garcia, N. Mirlean, P. R. Baisch, and E. B. Caram??o, “Assessment of polycyclic aromatic hydrocarbon influx and sediment contamination in an urbanized estuary,” Environ. Monit. Assess., vol. 168, no. 1–4, pp. 269–276, 2010. [11] R. J. Law, V. J. Dawes, R. J. Woodhead, and P. Matthiessen, “Polycyclic aromatic hydrocarbons (PAH) in seawater around England and Wales,” Mar. Pollut. Bull., vol. 34, no. 5, pp. 306–322, 1997. [12] G. WITT, “Polycyclic aromatic hydrocarbons in water and Sediment of the Baltic Sea,” Mar. Pollut. Bull., vol. 31, pp. 237–248, 1995. [13] R. van der Oost, J. Beyer, and N. P. E. Vermeulen, “Fish bioaccumulation and biomarkers in en v ironmental risk assessment : a re v iew,” Environ. Toxicol. Pharmacol., vol. 13, pp. 57–149, 2003. [14] R. M. Harrison, Pollution: Causes, Effects and Control, vol. 518. 2001. [15] R. . Gimeno, A. F. . Altelaar, R. . Marcé, and F. Borrull, “Determination of polycyclic aromatic hydrocarbons and polycylic aromatic sulfur heterocycles by high-performance liquid chromatography with fluorescence and atmospheric pressure chemical ionization mass spectrometry detection in seawater and sediment samp,” J. Chromatogr. A, vol. 958, no. 1, pp. 141–148, 2002. [16] R. . Marcé and F. Borrull, “Solid-phase extraction of polycyclic aromatic compounds,” J. Chromatogr. A, vol. 885, no. 1–2, pp. 273–290, 2000. [17] S. a. Wise, L. C. Sander, and M. M. Schantz, “Analytical Methods for Determination of Polycyclic Aromatic Hydrocarbons (PAHs) — A Historical Perspective on the 16 U.S. EPA Priority Pollutant PAHs,” Polycycl. Aromat. Compd., vol. 35, no. 2–4, pp. 187–247, 2015. [18] S. Jocis and F. Pierce, “Spectral Fluorescence Characteristics of Commercially Available Hydrocarbon Sensors United States Coast Guard Academy Faculty Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 22 - Advisor : Dr . Anni Vuorenkoski,” 2014. [19] Y. Li, S. Yoshida, Y. Chondo, H. Nassar, N. Tang, Y. Araki, A. Toriba, T. Kameda, and K. Hayakawa, “On-Line Concentration and Fluorescence Determination HPLC for Polycyclic Aromatic Hydrocarbons in Seawater Samples and Its Application to Japan Sea,” Chem. Pharm. Bull., vol. 60, no. April, pp. 531–535, 2012. [20] The Ocean Portal Team, Gulf Oil Spill, Smithsonian National Museum of Natural History, (2016), http://ocean.si.edu/gulf-oil-spill [21] L. Suberg, R. B. Wynn, J. Van Der Kooij, L. Fernand, S. Fielding, D. Guihen, D. Gillespie, M. Johnson, K. C. Gkikopoulou, I. J. Allan, B. Vrana, P. I. Miller, D. Smeed, and A. R. Jones, “Assessing the potential of autonomous submarine gliders for ecosystem monitoring across multiple trophic levels ( plankton to cetaceans ) and pollutants in shallow shelf seas,” Methods Oceanogr., vol. 10, no. July, pp. 70–89, 2014. Luis Aarón García Ortega Extraction system and determination of PAH in seawater and it implementation in submarine vehicles. - 23 - Memoria del TFG  Descripción detallada de las actividades desarrolladas durante la realización del TFT Las actividades consistieron en búsqueda bibliográfica para completar el contenido del TFT. Se realizaron presentaciones en power point con lo que se había avanzado y se expusieron en Inglés para por un lado desenvolverme con el inglés y por otro lado para que la tutora académica evaluara que es lo había hecho hasta la fecha y corregirme algunas cosas. A parte se realizó la redacción del TFT, búsqueda de figuras, realización de la presentación definitiva, traducción del TFT, etc.  Formación recibida (cursos, programas informáticos, etc.) No se recibió ningún tipo de formación adicional reglada, si se formó sobre el manejo del cromatógrafo líquido de alta resolución (HPLC) y del software de control para el procesado de los datos obtenidos.  Nivel de integración e implicación dentro del departamento y relaciones con el personal. La integración fue óptima en todo momento y la realización de labores se llevó a cabo en los tiempos estipulados. La relación con la tutora académica fue comunicativa así como su disposición fue total en todo momento.  Aspectos positivos y negativos más significativos relacionados con el desarrollo del TFT Como aspectos positivos cabe destacar el aprendizaje de habilidades como la escritura, la elaboración de un artículo, el procesado de datos, la mejora en el inglés redactado así como el contenido sobre el que versa el TFT. Como aspecto negativo destaco quizás que a lo largo de la carrera podrían hacer algo más de hincapié en la redacción de este tipo de artículos y en el uso del inglés en distintos ámbitos.  Valoración personal del aprendizaje conseguido a lo largo del TFT. Considero la valoración final como satisfactoria por lo enriquecedor de la experiencia así como por las habilidades y el conocimiento adquirido. La motivación fue mucha y la satisfacción bastante buena en general.