Development of methods for the determination of veterinary drugs in food matrices by liquid chromatography – mass spectrometry

The possible presence of veterinary drug residues and other contaminants in edible tissues and even food products is one of the key issues for food safety which arouses great public concern. The main objective of this thesis was the development of sensitive, selective, robust and effective analytical methods for the determination of veterinary drugs in food of animal origin using liquid chromatography hyphenated with mass spectrometric techniques. Η πιθανή παρουσία υπολειμμάτων κτηνιατρικών φαρμάκων και άλλων ρυπαντών σε βρώσιμους ιστούς και στα προϊόντα διατροφής είναι ένα από τα βασικά θέματα για την ασφάλεια των τροφίμων που προκαλεί μεγάλη ανησυχία στην κοινή γνώμη. Ο κύριος στόχος της παρούσας διπλωματικής εργασίας ήταν η ανάπτυξη ευαίσθητων, εκλεκτικών, ανθεκτικών και αποτελεσματικών αναλυτικών μεθόδων για τον προσδιορισμό κτηνιατρικών φαρμάκων σε τρόφιμα ζωικής προέλευσης με τη χρήση υγροχρωματογραφίας συζευγμένης με τεχνικές φασματομετρίας μάζας

Simultaneous determination of 148 pharmaceuticals and illicit drugs in sewage sludge based on ultrasound-assisted extraction and liquid chromatography–tandem mass spectrometry

This paper describes the development and validation of a new method for the simultaneous determination of 148 substances in sewage sludge. The selected compounds belong to different classes of pharmaceuticals, including antibiotics, analgesic and/or anti-inflammatory drugs, antiepileptics, benzodiazepines, antipsychotics, and antidepressants, among others, and illicit drugs, including opiates, opioids, cocaine, amphetamines, cannabinoids, and their metabolites. As far as the authors are aware, this is the first method in the peer-reviewed literature covering such a large number of target drugs for determination in a complex matrix like sewage sludge. The method presented herein combines ultrasoundassisted extraction (USE) and liquid chromatography coupled to tandem mass spectrometry. Good analytical performance was achieved, with limit-of-detection values below 10 ng g−1 d.w. for 91 % of the analytes and absolute recovery in the range 50–110 % for more than 77 % of the studied compounds. A combination of methanol and acidified water, also containing EDTA, proved to be the optimum solvent mixture to perform the extractions. An extra solid-phaseextraction clean-up step was not required, substantially reducing sample-preparation time and solvent consumption. Finally, the developed method was applied to the analysis of different sewage-sludge samples from five wastewater treatment plants of Santorini Island (Greece). Out of the 148 target compounds, 36 were detected. Several compounds, including acetylsalicylic acid, citalopram, and ciprofloxacin among others, had maximum concentrations above 100 ng g−1 d.w

Thorough investigation of the phenolic profile of reputable Greek honey varieties:varietal discrimination and floral markers identification using liquid chromatography–high-resolution mass spectrometry

Honey is a highly consumed commodity due to its potential health benefits upon certain consumption, resulting in a high market price. This fact indicates the need to protect honey from fraudulent acts by delivering comprehensive analytical methodologies. In this study, targeted, suspect and non-targeted metabolomic workflows were applied to identify botanical origin markers of Greek honey. Blossom honey samples (n = 62) and the unifloral fir (n = 10), oak (n = 24), pine (n = 39) and thyme (n = 34) honeys were analyzed using an ultra-high-performance liquid chromatography hybrid quadrupole time-of-flight mass spectrometry (UHPLC-q-TOF-MS) system. Several potential authenticity markers were revealed from the application of different metabolomic workflows. In detail, based on quantitative targeted analysis, three blossom honey markers were found, namely, galangin, pinocembrin and chrysin, while gallic acid concentration was found to be significantly higher in oak honey. Using suspect screening workflow, 12 additional bioactive compounds were identified and semi-quantified, achieving comprehensive metabolomic honey characterization. Lastly, by combining non-targeted screening with advanced chemometrics, it was possible to discriminate thyme from blossom honey and develop binary discriminatory models with high predictive power. In conclusion, a holistic approach to assessing the botanical origin of Greek honey is presented, highlighting the complementarity of the three applied metabolomic approaches

Ανάπτυξη μεθόδων προσδιορισμού κτηνιατρικών καταλοίπων σε τρόφιμα ζωικής προέλευσης με υγροχρωματογραφία – φασματομετρία μαζών

The possible presence of veterinary drug residues and other contaminants in edible tissues and even food products is one of the key issues for food safety which arouses great public concern. The main objective of this thesis was the development of sensitive, selective, robust and effective analytical methods for the determination of veterinary drugs in food of animal origin using liquid chromatography hyphenated with mass spectrometric techniques.Initially, an extended review of the veterinary drug classes and the existing methodologies for their determination is presented. The experimental section of the thesis is constituted of four parts: (1) Multi-residue determination of seventeen sulfonamides and five tetracyclines in fish tissue using a multi-stage LC-ESI-MS/MS approach based on advanced mass spectometric techniques (Chapter 3), (2) Qualitative multi-residue screening methods for 143 veterinary drugs and pharmaceuticals in milk and fish tissue using Liquid Chromatography Quadrupole-Time-Of-Flight Mass Spectrometry (Chapter 4), (3) Multi-residue determination of 115 veterinary drugs and pharmaceutical residues in milk powder, butter, fish tissue and eggs using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) (Chapter 5) and Multiresidue / Multiclass Determination Of 76 Veterinary Drugs And Pharmaceuticals In Bovine Muscle Tissue By Hydrophilic Interaction Liquid Chromatography-Tandem Mass Spectrometry (HILIC-MS/MS) (Chapter 6).It is our strong belief that these studies will constitute a step forward in multiresidue veterinary drug analysis, providing rapid and reliable analytical results.Η πιθανή παρουσία υπολειμμάτων κτηνιατρικών φαρμάκων και άλλων ρυπαντών σε βρώσιμους ιστούς και στα προϊόντα διατροφής είναι ένα από τα βασικά θέματα για την ασφάλεια των τροφίμων που προκαλεί μεγάλη ανησυχία στην κοινή γνώμη. Ο κύριος στόχος της παρούσας διπλωματικής εργασίας ήταν η ανάπτυξη ευαίσθητων, εκλεκτικών, ανθεκτικών και αποτελεσματικών αναλυτικών μεθόδων για τον προσδιορισμό κτηνιατρικών φαρμάκων σε τρόφιμα ζωικής προέλευσης με τη χρήση υγροχρωματογραφίας συζευγμένης με τεχνικές φασματομετρίας μάζας. Αρχικά παρουσιάζεται μια εκτεταμένη ανασκόπηση των κατηγοριών των κτηνιατρικών φαρμάκων και των υπάρχοντων μεθόδων για τον προσδιορισμό τους. Το πειραματικό μέρος της διατριβής αποτελείται από τέσσερα μέρη: (1) Ανάπτυξη ταχέων μεθόδων διαλογής (screening) για τον προσδιορισμό μη στοχευμένων-ενώσεων σε ζωικούς ιστούς με χρήση προηγμένων τεχνικών σάρωσης διαδοχικής φασματομετρίας μαζών συζευγμένης με υγροχρωματογραφία (Κεφάλαιο 3), (2) Ανάπτυξη πολυ - υπολειμματικής μεθόδου ταυτόχρονου προσδιορισμού 143 κτηνιατρικών φαρμάκων και φαρμακευτικών προϊόντων σε γάλα και ιστό ψαριού με LC-QTOF-MS (Κεφάλαιο 4), (3) Ανάπτυξη πολυ - υπολειμματικής μεθόδου ταυτόχρονου προσδιορισμού 115 κτηνιατρικών φαρμάκων και φαρμακευτικών καταλοίπων σε σκόνη γάλακτος, βούτυρο, ιστό ψαριού και αυγό χρησιμοποιώντας υγροχρωματογραφία υψηλής απόδοσης συζευγμένης με διαδοχική φασματομετρία μαζών (Κεφάλαιο 5) και Ανάπτυξη πολυ - υπολειμματικής μεθόδου προσδιορισμού 76 κτηνιατρικών καταλοίπων και φαρμακευτικών ουσιών σε ζωικούς ιστούς με υγροχρωματογραφία υδρόφιλων αλληλεπιδράσεων συζευγμένης με διαδοχική φασματομετρία μαζών (HILIC-MS/MS, Κεφάλαιο 6).Πεποίθησή μας είναι ότι οι μελέτες αυτές θα αποτελέσουν ένα βήμα προς τα εμπρός στην πολυ-υπολειμματική ανάλυση κτηνιατρικών φαρμάκων παρέχοντας γρήγορα και αξιόπιστα αναλυτικά αποτελέσματα

Quality and Authenticity Control of Fruit Juices-A Review

Food fraud, being the act of intentional adulteration of food for financial advantage, has vexed the consumers and the food industry throughout history. According to the European Committee on the Environment, Public Health and Food Safety, fruit juices are included in the top 10 food products that are most at risk of food fraud. Therefore, reliable, efficient, sensitive and cost-effective analytical methodologies need to be developed continuously to guarantee fruit juice quality and safety. This review covers the latest advances in the past ten years concerning the targeted and non-targeted methodologies that have been developed to assure fruit juice authenticity and to preclude adulteration. Emphasis is placed on the use of hyphenated techniques and on the constantly-growing role of MS-based metabolomics in fruit juice quality control area

Exploring Human Metabolome after Wine Intake—A Review

Wine has a rich history dating back to 2200 BC, originally recognized for its medicinal properties. Today, with the aid of advanced technologies like metabolomics and sophisticated analytical techniques, we have gained remarkable insights into the molecular-level changes induced by wine consumption in the human organism. This review embarks on a comprehensive exploration of the alterations in human metabolome associated with wine consumption. A great number of 51 studies from the last 25 years were reviewed; these studies systematically investigated shifts in metabolic profiles within blood, urine, and feces samples, encompassing both short-term and long-term studies of the consumption of wine and wine derivatives. Significant metabolic alterations were observed in a wide variety of metabolites belonging to different compound classes, such as phenolic compounds, lipids, organic acids, and amino acids, among others. Within these classes, both endogenous metabolites as well as diet-related metabolites that exhibited up-regulation or down-regulation following wine consumption were included. The up-regulation of short-chain fatty acids and the down-regulation of sphingomyelins after wine intake, as well as the up-regulation of gut microbial fermentation metabolites like vanillic and syringic acid are some of the most important findings reported in the reviewed literature. Our results confirm the intact passage of certain wine compounds, such as tartaric acid and other wine acids, to the human organism. In an era where the health effects of wine consumption are of growing interest, this review offers a holistic perspective on the metabolic underpinnings of this centuries-old tradition

Authentication of Greek PDO Kalamata Table Olives: A Novel Non-Target High Resolution Mass Spectrometric Approach

Food science continually requires the development of novel analytical methods to prevent fraudulent actions and guarantee food authenticity. Greek table olives, one of the most emblematic and valuable Greek national products, are often subjected to economically motivated fraud. In this work, a novel ultra-high-performance liquid chromatography–quadrupole time of flight tandem mass spectrometry (UHPLC-QTOF-MS) analytical method was developed to detect the mislabeling of Greek PDO Kalamata table olives, and thereby establish their authenticity. A non-targeted screening workflow was applied, coupled to advanced chemometric techniques such as Principal Component Analysis (PCA) and Partial Least Square Discriminant Analysis (PLS-DA) in order to fingerprint and accurately discriminate PDO Greek Kalamata olives from Kalamata (or Kalamon) type olives from Egypt and Chile. The method performance was evaluated using a target set of phenolic compounds and several validation parameters were calculated. Overall, 65 table olive samples from Greece, Egypt, and Chile were analyzed and processed for the model development and its accuracy was validated. The robustness of the chemometric model was tested using 11 Greek Kalamon olive samples that were produced during the following crop year, 2018, and they were successfully classified as Greek Kalamon olives from Kalamata. Twenty-six characteristic authenticity markers were indicated to be responsible for the discrimination of Kalamon olives of different geographical origins

Development and Validation of Pesticide Residues Determination Method in Fruits and Vegetables through Liquid and Gas Chromatography Tandem Mass Spectrometry (LC-MS/MS and GC-MS/MS) Employing Modified QuEChERS Method and a Centrifugal Vacuum Concentrator

Pesticides are used for controlling organisms, weeds and animals, causing damage to plants. Although the use of pesticides is a prerequisite for producing safe food, their accumulation makes their rapid determination necessary to avoid negative impacts on human health. The aim of this study was to develop reliable and robust analytical methods for the determination of pesticide residues in fruits and vegetables, validated according to SANTE/12682/2019 guidance. Five different categories of fruits and vegetables were selected (apple, orange, onion, lettuce, tomato). The sample preparation was based on QuEChERS methodology, slightly modified in the clean-up step, and appropriate d-SPE reagents were selected for each commodity. A Multi-Tube Vortexer was used for better agitation. In the final step, the extract was split in two: one part was acidified and injected in LC-MS/MS and the other part was evaporated in a centrifugal vacuum concentrator, and reconstituted and injected in GC-MS/MS. With the centrifugal vacuum concentrator used instead of nitrogen stream, more pesticides were determined, while sensitivity and repeatability increased. Validation results satisfied the SANTE/12682/2019 guidelines for approximately 220 analytes for each commodity. LOQ was set at 0.010 mg/kg for all analytes. Successful external quality assessment (proficiency testing) proved that the methods are fit for purpose