Roadmap of cocoa quality and authenticity control in the industry: a review of conventional and alternative methods

[EN] Cocoa (Theobroma cacao L.) and its derivatives are appreciated for their aroma, color, and healthy properties, and are commodities of high economic value worldwide. Wide ranges of conventional methods have been used for years to guarantee cocoa quality. Recently, however, demand for global cocoa and the requirements of sensory, functional, and safety cocoa attributes have changed. On the one hand, society and health authorities are increasingly demanding new more accurate quality control tests, including not only the analysis of physicochemical and sensory parameters, but also determinations of functional compounds and contaminants (some of which come in trace quantities). On the other hand, increased production forces industries to seek quality control techniques based on fast, nondestructive online methods. Finally, an increase in global cocoa demand and a consequent rise in prices can lead to future cases of fraud. For this reason, new analytes, technologies, and ways to analyze data are being researched, developed, and implemented into research or quality laboratories to control cocoa quality and authenticity. The main advances made in destructive techniques focus on developing new and more sensitive methods such as chromatographic analysis to detect metabolites and contaminants in trace quantities. These methods are used to assess cocoa quality; study new functional properties; control cocoa authenticity; or detect frequent emerging frauds. Regarding nondestructive methods, spectroscopy is the most explored technique, which is conducted within the near infrared range, and also within the medium infrared range to a lesser extent. It is applied mainly in the postharvest stage of cocoa beans to analyze different biochemical parameters or to assess the authenticity of cocoa and its derivatives.The authors wish to acknowledge the financial assistance provided by the Spanish Government and European Regional Development Fund (Project RTC-2016-5241-2). Maribel Quelal Vásconez thanks the Ministry Higher Education, Science, Technology, and Innovation (SENESCYT) of the Republic of Ecuador for her PhD grant.Quelal-Vásconez, MA.; Lerma-García, MJ.; Pérez-Esteve, É.; Talens Oliag, P.; Barat Baviera, JM. (2020). Roadmap of cocoa quality and authenticity control in the industry: a review of conventional and alternative methods. Comprehensive Reviews in Food Science and Food Safety. 19(2):448-478. https://doi.org/10.1111/1541-4337.12522S448478192Abdullahi, G., Muhamad, R., Dzolkhifli, O., & Sinniah, U. R. (2018). Analysis of quality retentions in cocoa beans exposed to solar heat treatment in cardboard solar heater box. 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Food forensics

The food market nowadays accounts for huge incomes and therefore it is an easy target for falsification. This recalls the urgency for reliable and powerful diagnostic techniques, in order to develop analytical protocols for identification of frauds. MS-based strategies of analysis are definitely suitable for this task and have become in the last years of paramount importance in the field of food forensics. Sophisticated techniques have been developed that request short times of analysis and allow the identification of specific parameters, useful as classification markers. The wide range of techniques available [i.e. isotopic analysis, inductively coupled plasma - mass spectrometry (ICP-MS), hyphenated systems, stand-alone systems] allow to address a wide range of analytical questions pertaining to food authentication and traceability

The Quality Control of Puerariae Lobatae Radix and Puerariae Thomsonii Radix

Puerariae Lobatae Radix (PLR) and Puerariae Thomsonii Radix (PTR) are traditional Chinese medicines used interchangeably in clinical practice, even though they possess significantly different chemical profiles. The aim of this thesis was to differentiate PLR from PTR using various analytical instruments coupled with chemometrics. Morphological results illustrate PLR possessed distinct macroscopic and microscopic features as compared to PTR. UPLC results reveal isoflavonoids were the major chemical constituents in both species, with the content of puerarin in PLR significantly greater than in PTR. PLS-DA models demonstrate both UPLC and HPTLC chromatographic fingerprints were effective in differentiating PLR from PTR. PLSR coupled with Raman spectra was able to predict the TPC and antioxidant capacities of PLR and PTR. The pharmacological results illustrate PLR possessed significantly greater anti-diabetic, cytoprotective and anti-cancer activities as compared to PTR. In summary, the results reveal the chemical fingerprints coupled with chemometrics was effective in differentiating PLR from PTR, and PLR was morphologically, chemically and pharmacologically different from PTR. This thesis provided further insight into the comprehensive nature of the quality control of two similar species and recommends changes to their descriptions in the pharmacopoeias. This will ultimately improve the quality, safety and efficacy of herbal products

NMR Metabolomics of Foods – Investigating the Influence of Origin on Sea Buckthorn Berries, Brassica Oilseeds and Honey

The origin of foods plays an important role in their metabolome (the set of compounds present as products of metabolic events). The compositions of food plants are inevitably determined by a number of inherent and external factors – most importantly by the genotype (species, subspecies, cultivar, variety) and the prevailing conditions and weather parameters at each growth environment. The declaration of food origin can be defined and protected by law. The constantly increasing consumer awareness towards food origin, authenticity and quality has set the need for efficient tools for their verification. Metabolomics based on nuclear magnetic resonance (NMR) spectroscopy is increasingly being applied in analysing food composition and quality and in detecting food frauds and adulterations. The aim of the current work was to determine the influence of origin-related variables in food composition and quality by using 1H NMR metabolomics. The model foods – sea buckthorn (Hippophaë rhamnoides) berries, oilseeds of Brassica spp. and varietal honey – represent different foods with special sensory, nutritional, bioactive, commercial and national significance. The sea buckthorn berry metabolites were investigated in respect to the genotype (subspecies, cultivar) and geographical origin, with special emphasis on the influence of northern latitudes and related conditions. In the oilseeds, the inter-species variation and the influence of environmental and developmental stage on the seed composition was investigated. NMR profiling was applied in characterising the marker compounds for different honey types for botanical authentication. Multivariate analysis methods such as principal component (PCA) and discriminant analyses (PLS-DA, OPLS-DA) were applied in every sub-study to determine the key metabolites and origin-related factors characterising the food samples. The sea buckthorn subspecies were mainly distinguished by the relatively high content of ethyl-β-D-glucopyranoside (ssp. rhamnoides) and malic acid and vitamin C (ssp. sinensis). The northern latitude and respective conditions (the length of growth season, temperature, radiation and precipitation) was shown to alter the chemical composition of berries of the same genetic origin. In subarctic latitudes, the berries formed more ascorbic acid while the levels of ethyl glucose remained relatively low. The berries of cultivar 'Tytti' contained more ethyl glucoside while the berries of 'Terhi' contained more quinic acid in comparison. Calculated from the start of the growing season until harvest, the effective temperature sum (degree days) and the radiation sum correlated positively with ethyl glucoside that accumulated up to six-fold in overripe berries in southern Finland. The sea buckthorn berries (ssp. sinensis) grown at over 2000 m altitude contained typically more ascorbic and malic acids. The seeds of turnip rape was characterised by a relatively higher sucrose and polyunsaturated fatty acid content over oilseed rape that had a higher content of sinapine and oil in general. Growth conditions with reduced temperature added to the level of unsaturation in the oilseed lipids and delayed the seed development. The varietal honeys were classified with the aid of NMR profiling, as the typical sugar composition and other botanical markers were characterised. Also, previously unreported markers were designated for dandelion honeys. The correlations between complex food metabolomes and the origin-related variables were easily accomplished with NMR metabolomics. Especially, the effect of northern conditions on the growth place-dependent compositional flexibility (phenotypic plasticity) of the plant foods was deemed considerable. The results of this thesis can be further used to determine food quality, origin and authenticity and as an aid in plant breeding operations.Alkuperällä on suuri vaikutus elintarvikkeen metabolomiin eli aineenvaihduntatuotteiden kokonaisuuteen. Erityisesti kasviperäisten elintarvikkeiden koostumukseen vaikuttavat lukuisat sisäiset ja ulkoiset alkuperään liittyvät tekijät, kuten perimä (laji, alalaji, lajike) ja kasvupaikalle tyypilliset ympäristö- ja sääolosuhteet. Elintarvikkeen alkuperä voidaan määritellä ja suojata lainsäädännöllisin perustein. Kuluttajien kasvanut kiinnostus ja tietämys elintarvikkeiden alkuperää, aitoutta ja laatua kohtaan on lisännyt tehokkaiden ja luotettavien laadunvarmistusmenetelmien tarvetta. Varsinkin ydinmagneettista resonanssispektroskopiaan (NMR) perustuvaa metabolomiikkatutkimusta hyödynnetään yhä useammin elintarvikkeiden koostumuksen, laadun ja aitouden analysoinnissa. Tämän tutkimuksen tarkoituksena oli selvittää alkuperän vaikutusta tyrnimarjojen (Hippophaë rhamnoides), rypsin- ja rapsinsiementen (Brassica spp.) sekä lajihunajan koostumukseen 1H-NMR-metabolomiikan avulla. Nämä elintarvikkeet ovat kansallisesti ja kaupallisesti arvokkaita ja mielenkiintoisia niille tyypillisten aistittavien, ravitsemuksellisten ja bioaktiivisten ominaisuuksien ansiosta. Tyrnimarjojen koostumusta vertailtiin eri alalajien (ssp. rhamnoides ja ssp. sinensis) ja lajikkeiden ('Terhi' ja 'Tytti') sekä kasvupaikkojen (Suomi, Kiina, Kanada) välillä. Tavoitteena oli erityisesti selvittää, miten erityisesti pohjoisille leveysasteille tyypilliset olosuhteet vaikuttavat marjojen aineenvaihduntatuotteisiin. Öljysiementen kohdalla tutkittiin myös miten lajikohtainen perimä sekä kasvupaikan/-olosuhteiden ja siemenen kehittymisvaihe vaikuttavat siementen kemialliseen koostumukseen ja laatuun. Hunajien tapauksessa NMR-metabolomiikkaa hyödynnettiin kasvialkuperäkohtaisten sormenjälkiyhdisteiden tunnistamiseen ja kotimaisten lajihunajien kasvialkuperän varmentamiseen. Kaikissa osatutkimuksissa sovellettiin pääkomponentti-(PCA) ja diskriminanttianalyysiin (PLS-DA, OPLS-DA) perustuvia monimuuttujamenetelmiä tärkeimpien näyteryhmiä erottavien ja määrittävien yhdisteiden ja taustatekijöiden selvittämiseksi. Tyrnin alalajit erottuivat pääasiassa suhteellisesti korkean etyyli-β-D-glukopyranosidin (ssp. rhamnoides) sekä omenahappo- ja C-vitamiini-pitoisuuden (ssp. sinensis) perusteella. Pohjoisen leveysasteen ja sille tyypillisten olosuhteiden (kasvukauden pituus, lämpötila, säteily, sademäärä) todettiin muokanneen samaa geneettistä alkuperää olevien marjojen kemiallista koostumusta. Subarktisilla leveyksillä tyrnimarjaan muodostui enemmän askorbiinihappoa ja etyyliglukosidin määrä oli alhainen. 'Tytti'-lajikkeen marjat sisälsivät enemmän etyyliglukosidia, kun taas 'Terhi' sisälsi vastaavasti enemmän kviinihappoa. Kasvukauden tehoisa lämpösumma ja säteilysumma korreloivat positiivisesti etyyliglukosidin kanssa, jota kertyi ylikypsiin marjoihin Etelä-Suomessa jopa kuusinkertainen määrä kypsiin verrattuna. Yli 2000 metrin korkeudessa kasvaneissa tyrnimarjoissa (ssp. sinensis) oli tyypillisesti korkeampi omena- ja askorbiinihappopitoisuus. Suhteellisesti korkeampi sakkaroosipitoisuus ja monityydyttymättömien rasvahappojen osuus oli tyypillisempää rypsille, kun taas rapsi erottui rypsistä korkeamman öljy- ja sinapiinipitoisuuden perusteella. Kylmempi kasvupaikka lisäsi monityydyttymättömien rasvahappojen osuutta öljysiemenissä ja hidasti siemenen kehittymistä. NMR-profiloinnin avulla lajihunajat pystyttiin luokittelemaan kullekin hunajalle ominaisen sokerikoostumuksen ja muiden kasvialkuperästä kertovien merkkiyhdisteiden perusteella. Voikukkahunajalle tunnistettiin myös aiemmin raportoimattomia merkkiyhdisteitä. NMR-metabolomiikan avulla pystyttiin helposti selvittämään monimutkaisten aineenvaihduntatuotteiden kokonaisuuksien ja elintarvikkeen alkuperään liittyvien muuttujien välisiä yhteyksiä. Varsinkin pohjoisten kasvuolosuhteiden vaikutus kasviperäisten elintarvikkeiden koostumukselliseen vaihteluun oli huomattava. Väitöskirjan tuloksia voidaan hyödyntää elintarvikkeiden laadun, alkuperän ja aitouden varmistamisessa sekä kasvinjalostuksen apuna.Siirretty Doriast

Kinetics of environmental biocomplexity : experiments, quantum chemistry and machine learning

Tese (doutorado) — Universidade de Brasília, Instituto de Química, Programa de Pós-Graduação em Química, 2022.Micro poluentes de preocupação emergente têm imposto um grande desafio tecnológico: pesticidas, drogas e outras substâncias antropogênicas são cada vez mais encontrados em ambientes aquáticos e atmosféricos e até mesmo no abastecimento de água, estando relacionados a efeitos adversos sobre a biota e a saúde humana. Superar esse desafio requer a compreensão do comportamento dessas espécies no meio ambiente e o desenvolvimento de tecnologias que permitam minimizar sua disseminação. Alternativas viáveis aplicadas nesta tese incluem o uso de processos de oxidação baseado em radicais utilizando tanto o método experimental – através do método cinético de competição – quanto os protocolos teóricos – um conjunto de cálculos cinéticos, quânticos e aprendizado de máquina. Em um primeiro estudo, os mecanismos, cinéticas e uma avaliação da toxicidade da degradação do picloram – pesticida amplamente utilizado no mundo – iniciados por radicais OH indicam que: i) duas vias favoráveis ocorrem por adição ao anel de piridina, ii) picloram e a maioria dos produtos de degradação são estimados como prejudiciais; no entanto, ii) esses compostos podem sofrer fotólise pela luz solar. No entanto, o método cinético da competição e a descrição da química quântica fazem da degradação uma empreendimento formidável, considerando os custos de equipamentos instrumentais ad hoc e esforços computacionais dedicados. Para superar os exigentes procedimentos convencionais, desenvolvemos uma aplicação web gratuita e de fácil acesso (www.pysirc.com.br) baseada no aprendizado de máquina holístico combinado com modelos de impressões digitais moleculares que permitem a compilação de parâmetros cinéticos e interpretação mecanicista de ataques de oxidação baseado em radicais de acordo com os princípios da OCDE. Algoritmos de aprendizagem de máquina foram implementados, e todos os modelos forneceram alto desempenho de ajuste para a degradação baseado em radical no ambiente aquático e atmosférico. Os modelos foram interpretados utilizando o método SHAP (Explicações Aditivas de SHapley): os resultados mostraram que o modelo desenvolvido fez a previsão com base em uma compreensão razoável de como grupos de retirada/doação de elétrons interferem na reatividade dos radicais. Argumentamos que nossos modelos e interface web podem estimular e expandir a aplicação e interpretação de pesquisas cinéticas sobre contaminantes em unidades de tratamento de água e ar com base em tecnologias oxidativas avançadas.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES); Fundação de Apoio à Pesquisa do Distrito Federal (FAP/DF) e Fundação de Amparo à Pesquisa do Estado de Goiás (FAPEG).Micro-pollutants of emerging concern have imposed a major technological challenge: pesticides, drugs and other anthropogenic substances are increasingly found in aquatic and atmospheric environments and even in water supplies, being related to adverse effects on biota and human health. Overcoming this challenge requires understanding the behavior of these species in the environment and the development of technologies that allows for minimizing their dissemination. Viable alternatives applied in this thesis include the use of radical-based oxidation processes using both experimental – via the competition kinetics method – and theoretical protocols – blend of kinetic, quantum chemistry and machine learning calculations. In a first study, the mechanisms, kinetics, and an evaluation of the toxicity of picloram degradation – a pesticide widely used in the world - initiated by OH radicals indicate that: i) two favorable pathways occur by addition to the pyridine ring, ii) picloram and the majority of degradation products are estimated as harmful; however, ii) these compounds can suffer photolysis by sunlight. However, the competition kinetic method and the quantum chemistry description make the degradation analyses a formidable enterprise, considering the costs of ad hoc instrumental equipment’s and dedicated computational efforts. To overcome the demanding conventional procedures, we developed a free and user-friendly web application (www.pysirc.com.br) based on holistic machine learning combined with molecular fingerprints models that permits compilation of kinetic parameters and mechanistic interpretation of radical-based oxidation attacks according to the OECD principles. Machine learning algorithms were implemented, and all models provided high goodness-of-fit for radical-based degradation in aquatic and atmospheric environment. The models were interpreted using the SHAP (SHapley Additive exPlanations) method: the results showed that the model developed made the prediction based on a reasonable understanding of how electron-withdrawing/donating groups interfere in the reactivity of the radicals. We argue that our models and web interface can stimulate and expand the application and interpretation of kinetic research on contaminants in water and air treatment units based on advanced oxidative technologies

Application of Infrared Spectroscopy and Chemometrics to the Cocoa Industry for Fast Composition Analysis and Fraud Detection

Tesis por compendio[ES] El cacao es un producto de alto valor, no únicamente por sus características sensoriales, sino porque también presenta un alto contenido en antioxidantes y alcaloides estimulantes con efectos saludables. Debido a la alta demanda, la industria del cacao en polvo tiene el desafío de asegurar la calidad de grandes volúmenes de producción de una manera rápida y precisa, evitando la presencia de contaminantes o adulterantes en la materia prima, ofreciendo productos donde se preserven las propiedades saludables. La espectroscopia del infrarrojo cercano (NIR) es una tecnología rápida y no destructiva útil en el análisis de productos alimentarios. La presente tesis doctoral se centra en evaluar el potencial uso del NIR como una herramienta de control de calidad con el fin de poder resolver problemas que se presentan en la industria del cacao en polvo. Los problemas a resolver incluyen la detección de materiales no deseados o adulterantes en el cacao en polvo, y la monitorización rápida y precisa del contenido de flavanoles y metilxantinas del cacao en polvo durante el proceso de alcalinización. El primer capítulo evalúa la viabilidad del NIR, en combinación con análisis quimiométricos, en la detección de la presencia de materiales no deseados o adulterantes como son cascarilla de cacao o harina de algarroba. Para ello, diferentes muestras de cacao en polvo natural y con diferentes niveles de alcalinización (suave, medio y fuerte) fueron mezcladas con distintas proporciones de cascarilla de cacao (con cacao natural) o harina de algarroba (con cacao natural y alcalinizado). Los resultados obtenidos indican que el NIR, combinado con modelos estadísticos tales como el análisis discriminante por mínimos cuadrados parciales (PLS-DA) y la regresión parcial de mínimos cuadrados (PLS), es un método rápido y eficaz para identificar cualitativa y cuantitativamente materiales no deseados o adulterantes como la cascarilla y la algarroba en cacao en polvo, independientemente del grado de alcalinización o el nivel de tostado de la harina de algarroba. En el segundo capítulo, el análisis composicional del cacao en polvo se orientó al control de los cambios producidos en el contenido de flavanoles y metilxantinas debidos al proceso de alcalinización al que se somete el caco en polvo. Se determinó el contenido de catequina, epicatequina, cafeína y teobromina mediante cromatografía líquida de alta resolución (HPLC), correlacionándose los contenidos obtenidos para cada uno de estos compuestos con las determinaciones NIR. Se obtuvieron buenos modelos para la predicción de los compuestos mediante regresión PLS con valores superiores a 3 para la relación entre el rendimiento y la desviación (RDP), lo cual demuestra que los modelos obtenidos pueden ser utilizados para la rápida y fiable predicción del contenido de flavanoles y metilxantinas en cacaos naturales y con diferentes niveles de alcalinización.[CA] El cacau és un producte d'alt valor, no sols per les seues característiques sensorials, sinó perquè també presenta un elevat contingut en antioxidants i alcaloids estimulants amb efectes saludables. A conseqüència a l'alta demanda, l'industria del cacau en pols té el desafiament d'assegurar la qualitat de grans volums de producció d'una manera ràpida i precisa, evitant la presència de contaminants o adulterants en la matèria cosina, oferint productes a on se preserven les propietats saludables. L'espectroscòpia de l'infrarroig proper (NIR) és una tecnologia ràpida i no destructiva útil en l'anàlisi de productes alimentaris. La present tesis doctoral se centra en avaluar el potencial ús del NIR com una eina de control de qualitat amb l'objectiu de poder resoldre problemes que es presenten en l'industria del cacau en pols. Els problemes a resoldre inclouen la detecció de materials no desitjats o adulterants en el cacau en pols, i la monitorització ràpida i precisa del contingut de flavanols i metilxantines del cacau en pols durant el procés d'alcalinització. El primer capítol avalua la viabilitat del NIR, en combinació amb anàlisis quimiométrics, en la detecció de la presència de materials no desitjats o adulterants com són pellofa de cacau o farina de garrofa. Per a això, diferents mostres de cacau en pols natural i amb diferents nivells d'alcalinització (suau, mig i fort) foren barrejades en distintes proporcions de pellofa de cacau (en cacau natural) o farina de garrofa (en cacau natural i alcalinisat). Els resultats obtinguts per a NIR, combinats amb models estadístics com l'anàlisi discriminant per mínims quadrats parcials (PLS-DA) i la regressió parcial de mínims quadrats (PLS), és un mètode ràpid i eficaç per identificar materials no desitjats o adulterants com la pellofa de cacau o la farina de garrofa, amb independència del grau d'alcalinització del cacau o de torrat de la farina de garrofa. En el segon capítol, l'anàlisi composicional del cacau en pols s'orientà al control dels canvis produïts en el contingut de flavanols i metilxantines a causa del procés d'alcalinització al que se sotmet el cacau en pols. Es va determinar el contingut de catequina, epicatequina, cafeïna i teobromina mitjançant cromatografia líquida d'alta resolució (HPLC), i es van correlacionar els continguts obtinguts per a cadascun d'estos composts amb les determinacions NIR. Es van obtindré bons models per a la predicció dels composts mitjançant regressió PLS amb valors superiors a 3 per a la relació entre el rendiment i la desviació (RDP), la qual cosa demostra que els models obtinguts poden ser emprats per a la ràpida i fiable predicció del contingut de flavanols i metilxantines en cacaus naturals o amb diferents nivells d'alcalinització.[EN] Cocoa is a product of high value, not only because of its sensory characteristics, but also because it has a high content of antioxidants and stimulating alkaloids with health effects. Due to the high demand, the cocoa powder industry has the challenge of ensuring the quality of large volumes of production in a fast and accurate way, avoiding the presence of contaminants or adulterants in the raw material, offering products where the healthy properties are preserved. The near infrared spectroscopy (NIR) is a rapid and non-destructive technology useful in the analysis of food products. The present doctoral thesis focuses on evaluating the potential use of NIR as a quality control tool in order to solve problems that arise in the cocoa industry powdered. The problems to solve include the detection of unwanted materials or adulterants in the cocoa powder, and the rapid and accurate monitorization of the flavanols and methylxanthines content in the cocoa powder during the alkalization process. The first chapter evaluates the viability of the NIR, in combination with chemometric analysis, in the detection of presence of unwanted materials or adulterants such as cocoa shell or carob flour. For this, different samples of natural cocoa powder and with different levels of alkalization (light, medium and strong) were mixed with different proportions of cocoa shell (with natural cocoa) or carob flour (with natural and alkalized cocoa). The results obtained indicate that the NIR combined with statistical models such as the partial least squares discriminant analysis (PLS-DA) and the partial least squares regression (PLS), is a fast and efficient method to identify qualitative and quantitative unwanted materials or adulterants such as shell and carob in cocoa powder, regardless of the degree of alkalization or level of roasting of carob flour. In the second chapter, the compositional analysis of cocoa powder was oriented to the control of the changes produced in the content of flavanols and methylxanthines due to the process of alkalization to which the cocoa powder is subjected. The content of catechin, epicatechin, caffeine and theobromine were determined by high performance liquid chromatography (HPLC), correlating the contents obtained for each of these compounds with the NIR determinations. Good models were obtained for the prediction of compounds by regression PLS with values above 3 for the ratio of performance to deviation (RDP), which shows that the obtained models can be used for the quick and reliable prediction of flavanol content and methylxanthines in natural cocoas and with different alkalization levels.This Doctoral Thesis has been carried out thanks to a doctoral studies scholarship granted by the Ministry of Higher Education, Science, Technology and Innovation (SENESCYT) of the Republic of EcuadorQuelal Vásconez, MA. (2019). Application of Infrared Spectroscopy and Chemometrics to the Cocoa Industry for Fast Composition Analysis and Fraud Detection [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/135258TESISCompendi