ΠΟΙΟΤΙΚΗ ΚΑΙ ΠΟΣΟΤΙΚΗ ΧΗΜΙΚΗ ΑΝΑΛΥΣΗ ΕΠΙΛΕΓΜΕΝΩΝ ΔΕΙΓΜΑΤΩΝ ΑΙΘΕΡΙΩΝ ΕΛΑΙΩΝ ΛΕΒΑΝΤΑΣ ΕΛΛΗΝΙΚΗΣ ΠΡΟΕΛΕΥΣΕΩΣ: ΑΞΙΟΛΟΓΗΣΗ ΒΑΣΕΙ ΤΩΝ ΠΡΟΔΙΑΓΡΑΦΩΝ ΤΗΣ ΕΥΤΩΠΑΪΚΗΣ ΦΑΡΜΑΚΟΠΟΙΙΑΣ

Στην παρούσα πτυχιακή εργασία πραγματοποιήθηκε ποιοτική ανάλυση και προσδιορισμός της ποσοστιαίας σύστασης επιλεγμένων δειγμάτων αιθέριων ελαίων λεβάντας ελληνικής προελεύσεως. Στη συνέχεια έγινε σύγκριση και αξιολόγηση των ποσοτικών αποτελεσμάτων με τις προδιαγραφές που ορίζει η Ευρωπαϊκή Φαρμακοποιία για τη φαρμακευτική χρήση του αιθέριου ελαίου της λεβάντας. Για όλες τις μετρήσεις που πραγματοποιήθηκαν σε αυτή την εργασία, χρησιμοποιήθηκε αεριοχρωματογράφος Hewlett-Packard 6890, συνδεδεμένος με σύστημα φασματομέτρου μαζών Hewlett-Packard 5973 που λειτουργεί με ιοντισμό με ηλεκτροψεκασμό (ESI) και με ενέργεια ιοντισμού 70 eV. Η ταυτοποίηση των πτητικών συστατικών επιτεύχθηκε με τη χρήση τη βιβλιοθήκης δεδομένων Wiley275, βιβλιογραφικών δεδομένων και με επιμέρους μελέτη των θραυσμάτων της φασματομετρίας μαζών. Ελέγχοντας τις απαιτήσεις της Ευρωπαϊκής Φαρμακοποιίας και συγκρίνοντάς τις με τα δεδομένα από τις αναλύσεις που πραγματοποιήθηκαν, παρατηρείται πως τα περισσότερα από τα υπό εξέταση δείγματα αιθέριων ελαίων λεβάντας ικανοποιούν σε μεγάλο βαθμό τις προδιαγραφές της, με εξαίρεση κάποια δείγματα, τα οποία αποκλίνουν σε κάποια σημεία από τις επιθυμητές τιμές.In this thesis, a qualitative analysis and determination of the percentage composition of selected samples of Greek lavender essential oils was conducted, in order to compare and evaluate the quantitative results with the standards set by the European Pharmacopoeia for pharmaceutical use of the essential oil of lavender. For all measurements made in this paper, we used gas-chromatographer Hewlett-Packard 6890, connected with mass spectrometer Hewlett-Packard 5973, that works by using electrospray ionisation (ESI) and ionization energy 70 eV. The identification of the volatile components was achieved by using the library Wiley275, bibliographical data and individual study of the fragments of mass spectrometry. After evaluating the standards of the European Pharmacopoeia and comparing them to data from the analysis carried out, it is observed that most of the test samples of essential lavender oil satisfy the standards, with the exception of some samples, which differ in some respects from the desired values

Disposition of two highly permeable drugs in the stomach and the upper small intestine of healthy adults after a standard high-calorie, high-fat meal

Objectives: To quantify the presence of two model highly permeable drugs, paracetamol and danazol, in the upper gastrointestinal lumen under conditions simulating the situation after disintegration of immediate release dosage forms administered in bioavailability/bioequivalence studies in the fed state. To understand the drug transfer process from the antral contents through the upper small intestine using the luminal drug data. Methods: 8 healthy male adult volunteers participated in a randomized, single dose, two-phase, crossover study. After evaluating the impact of homogenization on meal’s viscosity and particle size, the meal, containing phenol red as non-absorbable marker, was administered to the antrum via the gastric lumen of a naso-gastro-intestinal tube. The drugs were administered in solution form (Phase I) and in suspension form (Phase II) with a glass of tap water to the antrum of the stomach, 30 min after the initiation of meal administration. Samples were aspirated from the antrum and the upper small intestine up to 4 hours post drug administration. Results: Apparent concentrations in the aqueous contents of the antrum were higher than apparent concentrations in micellar contents of the upper small intestine for paracetamol; the opposite was observed for danazol. Based on total drug amount per volume data in contents of the upper gastrointestinal lumen, the transfer of paracetamol (aqueous solution or suspension) and danazol (aqueous suspension) through the upper small intestine could be described as an apparent first-order process. Transfer of a long-chain triglyceride solution of danazol was highly variable. Conclusions: Concentrations in the aqueous/micellar phase of luminal contents and values of parameters controlling the transfer from bulk gastric contents through the upper small intestine after a high-calorie, high-fat meal, were reported for the first time for highly permeable drugs. Data are expected to enhance the development of biorelevant in vitro and physiologically based biopharmaceutics modelling methodologies

THE IMPACT OF FOOD INTAKE ON THE LUMINAL ENVIRONMENT AND PERFORMANCE OF ORAL DRUG PRODUCTS WITH A VIEW TO IN VITRO AND IN SILICO SIMULATIONS: A PEARRL REVIEW

Objective: Using the type of meal and dosing conditions suggested by regulatory agencies as a basis, this review has two specific objectives. First, to summarize our understanding on the impact of food intake on luminal environment and drug product performance. Second, to summarize the usefulness and limitations of available in vitro and in silico methodologies for the evaluation of drug products performance after food intake. Key findings: Characterization of the luminal environment and studies evaluating product performance in the lumen, under conditions suggested by regulatory agencies for simulating the fed state, are limited. Various in vitro methodologies have been proposed for evaluating drug product performance in the fed state but systematic validation is lacking. Physiologically based pharmacokinetic (PBPK) modelling approaches require the use of in vitro biorelevant data and, to date, have been used primarily for investigating the mechanisms via which an already observed food effect is mediated. Conclusion: Better understanding of the impact of changes induced by the meal administration conditions suggested by regulatory agencies on the luminal fate of the drug product is needed. Relevant information will be useful for optimizing the in vitro test methods, and increasing the usefulness of PBPK modelling methodologies

In vitro simulation of the environment in the upper gastrointestinal lumen after drug administration in the fed state using the TIM-1 system and comparison with luminal data in adults

We evaluated the environment in TIM-1 luminal compartments using paracetamol and danazol solutions and suspensions and the fed state configuration. Data were compared with recently published data in healthy adults. TIM-1 Experiments were performed with a 3-fold downscale. Volumes of secretions in gastric and duodenal compartments adequately reflected the luminal data in adults up to 3h post drug dosing. pH values in duodenal and jejunal compartments adequately reflected average pH values in adults. In gastric compartment pH values where initially higher than average values in adults and reached baseline levels earlier than in adults. The environment in the TIM-1 gastric compartment and jejunal compartment adequately reflected the average total paracetamol and danazol amounts per volume of contents in the adult stomach and upper small intestine, respectively. Total bile acids concentrations in the micellar phase of contents in duodenal and jejunal compartments overestimated micellar concentrations in the upper small intestine of adults. Adjustments in gastric emptying / acid secretion rates and bile acids identities in the duodenal and jejunal compartments, and application of dynamic bile acids secretion rates are expected to further improve the relevance of luminal conditions in TIM-1 compartments with those in adults

Impact of regional differences along the gastrointestinal tract of healthy adults on oral drug absorption : an UNGAP review

Oral administration is the most common route of drug delivery. The absorption of a drug from the gut into the bloodstream involves disintegration of the solid dosage form, dissolution of the active pharmaceutical ingredient and its transport across the gut wall. The efficiency of these processes is determined by highly complex and dynamic interplay between the gastrointestinal tract, the dosage form and the API. The European Network on Understanding Gastrointestinal Absorption-related Processes (UNGAP)aims to improve our understanding of intestinal drug absorption by creating a multidisciplinary Network of researchers from academia and industry engaging in scientific discussions. As part of the basis for the UNGAP project, this review aims to summarize the current knowledge on anatomy and physiology of the human gastrointestinal tract with emphasis on human studies for the evaluation of the regional drug absorption and the prediction of oral dosage form performance. A range of factors and methods will be considered, including imaging methods, intraluminal sampling and, models for predicting segmental/regional absorption. In addition, in vitro and in silico methods to evaluate regional drug absorption will be discussed. This will provide the basis for further work on improving predictions for the in vivo behavior of drug products in the gastrointestinal tract

Impact of gastrointestinal tract variability on oral drug absorption and pharmacokinetics : an UNGAP review

The absorption of oral drugs is frequently plagued by significant variability with potentially serious therapeutic consequences. The source of variability can be traced back to interindividual variability in physiology, differences in special populations (age- and disease-dependent), drug and formulation properties, or food-drug interactions. Clinical evidence for the impact of some of these factors on drug pharmacokinetic variability is mounting: e.g. gastric pH and emptying time, small intestinal fluid properties, differences in pediatrics and the elderly, and surgical changes in gastrointestinal anatomy. However, the link of colonic factors variability (transit time, fluid composition, microbiome), sex differences (male vs. female) and gut-related diseases (chronic constipation, anorexia and cachexia) to drug absorption variability has not been firmly established yet. At the same time, a way to decrease oral drug pharmacokinetic variability is provided by the pharmaceutical industry: clinical evidence suggests that formulation approaches employed during drug development can decrease the variability in oral exposure. This review outlines the main drivers of oral drug exposure variability and potential approaches to overcome them, while highlighting existing knowledge gaps and guiding future studies in this area

Drug disposition and characteristics of contents in the upper gastrointestinal lumen of healthy adults after a standard high-calorie high fat meal – Implications for the in vitro drug testing conditions

Στόχοι Να ποσοτικοποιηθεί η παρουσία στον ανώτερο γαστρεντερικό αυλό και να κατανοηθεί η διαδικασία της γαστρεντερικής μεταφοράς φαρμάκων με υψηλή εντερική διαπερατότητα, σε συνθήκες που προσομοιώνουν την κατάσταση μετά την αποσάθρωση φαρμακευτικών προϊόντων άμεσης αποδέσμευσης, στις μελέτες βιοδιαθεσιμότητας/βιοϊσοδυναμίας κατά την περίοδο πέψης. Να εκτιμηθούν τα φυσικοχημικά χαρακτηριστικά των περιεχομένων του στομάχου και του ανώτερου λεπτού εντέρου και των αντιστοίχων υδατικών και μικκυλιακών φάσεων, μετά από τη χορήγηση του γεύματος που χρησιμοποιείται για να επάγει την περίοδο πέψης στις μελέτες βιοδιαθεσιμότητας/βιοϊσοδυναμίας (πρότυπο γεύμα). Να αξιολογηθεί η ανάγκη τροποποιήσεων σε in vitro μεθοδολογίες που χρησιμοποιούνται σήμερα για την εκτίμηση της ενδοαυλικής συμπεριφοράς per os χορηγούμενων προϊόντων στη διάρκεια της πέψης και συγκεκριμένα: (α) να αξιολογηθεί η επάρκεια της προσομοίωσης των ενδοαυλικών χαρακτηριστικών με τα υγρά μέσα που χρησιμοποιούνται σήμερα στις in vitro δοκιμασίες εκτίμησης της ενδοαυλικής συμπεριφοράς per os χορηγούμενων φαρμάκων, και (β) να εκτιμηθεί η επάρκεια της in vitro προσομοίωσης του ενδοαυλικού περιβάλλοντος με τη χρήση της διάταξης ΤΙΜ-1. Μέθοδοι Οκτώ υγιείς ενήλικοι άνδρες εθελοντές συμμετείχαν σε μια τυχαιοποιημένη, απλής δόσης, δύο φάσεων, διασταυρωτή μελέτη. Το πρότυπο γεύμα, που περιείχε τον μη απορροφούμενο δείκτη ερυθρό της φαινόλης, χορηγήθηκε στο άντρο μέσω ενός ρινο-γαστρο-δωδεκαδακτυλικού σωλήνα. Τα φάρμακα, παρακεταμόλη και δαναζόλη, χορηγήθηκαν υπό μορφή διαλυμάτων (Φάση Ι) και εναιωρημάτων (Φάση ΙΙ) με ένα ποτήρι νερό, 30 min μετά την έναρξη χορήγησης του γεύματος. Δείγματα αναρροφήθηκαν από το άντρο και το ανώτερο λεπτό έντερο για τις επόμενες τέσσερις ώρες. Τα δείγματα αναλύθηκαν για το περιεχόμενο τους σε φάρμακα, ερυθρό της φαινόλης, pH, ρυθμιστική χωρητικότητα, ιξώδες, ωσμωτικότητα, και παρουσία διαλυτοποιητικών παραγόντων. Στα πειράματα με τη διάταξη ΤΙΜ-1 προσομοιώθηκε το πρωτόκολλο της κλινικής μελέτης. Αποτελέσματα Οι φαινόμενες συγκεντρώσεις στην υδατική φάση των γαστρικών περιεχομένων ήταν υψηλότερες από αυτές στη μικκυλιακή φάση των περιεχομένων του ανώτερου λεπτού εντέρου για την παρακεταμόλη. Το αντίθετο παρατηρήθηκε για τη δαναζόλη. Η γαστρεντερική μεταφορά της παρακεταμόλης (υδατικό διάλυμα ή εναιώρημα) και της δαναζόλης (υδατικό εναιώρημα) θα μπορούσε να περιγραφεί ως μια φαινόμενη πρωτοταξική διαδικασία. Η μεταφορά του λιπιδικού διαλύματος δαναζόλης παρουσίασε πολύ υψηλή διατομική μεταβλητότητα. Το ιξώδες των περιεχομένων του ανώτερου γαστρεντερικού αυλού ήταν πολύ υψηλότερο από τις τιμές που έχουν αναφερθεί για τη διαπεπτική περίοδο. Προτάθηκαν νέα μέσα για την προσομοίωση των ενδογαστρικών περιεχομένων και της υδατικής φάσης των ενδογαστρικών περιεχομένων. Η διάταξη ΤΙΜ-1 προσομοίωσε ικανοποιητικά την παρουσία της παρακεταμόλης (διάλυμα και εναιώρημα) και της δαναζόλης (εναιώρημα) στον ανώτερο γαστρεντερικό αυλό, όχι όμως τις εκκρίσεις στο στόμαχο, την πέψη των λιπών και τα επίπεδα χολικών οξέων στο δωδεκαδάκτυλο. Συμπεράσματα Οι συγκεντρώσεις στα περιεχόμενα και στην υδατική/μικκυλιακή φάση των ενδοαυλικών περιεχομένων και οι τιμές των παραμέτρων που διέπουν την κινητική της γαστρεντερικής μεταφοράς των φαρμάκων μετά τη χορήγηση του προτύπου γεύματος περιεγράφηκαν για πρώτη φορά. Αντιπροσωπευτικές τιμές ιξώδους για τα περιεχόμενα του στομάχου και του ανωτέρου λεπτού εντέρου, καθώς και η σύσταση μέσων για την Επιπέδου ΙΙ προσομοίωση της υδατικής φάσης των γαστρικών περιεχομένων, μετά το πρότυπο γεύμα, προτάθηκαν για πρώτη φορά. Τα μέσα προσομοίωσης των ενδογαστρικών περιεχομένων στην περίοδο πέψης θα μπορούσαν να απλοποιηθούν, ενώ η σύσταση του μέσου FeSSIF-V2 επιβεβαιώθηκε ως αντιπροσωπευτική της μικυλλιακής φάσης των περιεχομένων του ανώτερου λεπτού εντέρου. Η διάταξη ΤΙΜ-1 ίσως αποτελέσει χρήσιμο εργαλείο για πολλές κατηγορίες φαρμάκων στο μέλλον εφόσον υπάρξουν βελτιστοποιήσεις στις συνθήκες που επικρατούν στο γαστρικό και στο νηστιδικό διαμέρισμα.Objectives To quantify the presence in the upper gastrointestinal lumen and understand the gastrointestinal transfer process of highly permeable drugs under conditions simulating the situation after disintegration of immediate release dosage forms administered in bioavailability/bioequivalence studies in the fed state. To measure the physicochemical characteristics of gastric contents and contents of the upper small intestine and their respective aqueous and micellar phases, after administration of the meal used to induce fed state conditions in bioequivalence/bioavailability studies (standard meal). To evaluate the need for adjustments in in vitro methodologies used to date to assess intraluminal behavior of per os administered drug products in the fed state and specifically: (a) to evaluate the sufficiency of simulation of intraluminal characteristics with liquid media used to date in in vitro testing of intraluminal behavior of per os administered drugs, and (b) to evaluate the competency of the in vitro simulation of the intraluminal environment using the TIM-1 model. Methods Eight healthy male adult volunteers participated in a randomized, single dose, two-phase, crossover study. The standard meal, containing the non-absorbable marker phenol red, was administered to the antrum via a naso-gastro-intestinal tube. The drugs, paracetamol and danazol, were administered in solution form (Phase I) and in suspension form (Phase II) with a glass of tap water, 30 min after initiation of the meal. Samples were aspirated from the antrum and the upper small intestine over the next four hours. Samples were analyzed for drug content, phenol red content, pH, buffer capacity, viscosity, osmolality, and presence of solubilizing agents. The clinical study protocol was mimicked in TIM-1 experiments. Results Apparent concentrations in the aqueous phase of antral contents were higher than apparent concentrations in the micellar phase of upper small intestinal contents for paracetamol. The opposite was observed for danazol. The gastrointestinal transfer of paracetamol (aqueous solution or suspension) and danazol (aqueous suspension) could be described as an apparent first-order process. The transfer of the long-chain triglyceride solution of danazol presented high intersubject variability. The viscosity of contents from the upper gastrointestinal lumen in the fed state was much higher than values reported in the fasted state. New biorelevant media for the simulation of intragastric contents and the aqueous phase of intragastric contents were suggested. The TIM-1 model simulated adequately the presence of paracetamol (solution or suspension) and danazol (suspension) in the upper gastrointestinal lumen, but not the gastric secretions, the digestion of lipids, and the bile acid levels. Conclusions Concentrations in the aqueous/micellar phase of luminal contents and values of parameters controlling drug transfer after the standard meal were reported for the first time. Representative viscosity values in the stomach and the upper small intestine and Level II composition of the aqueous phase of gastric contents, after the standard meal, were proposed for the first time. Biorelevant media simulating the intragastric conditions after ingestion of a standard meal could be simplified whereas FeSSIF-V2 composition was confirmed to be representative of the micellar phase of contents of the upper small intestine. The TIM-1 model could be a useful tool in drug development in the future after optimization of conditions in the gastric and jejunal compartments

The impact of food intake on the luminal environment and performance of oral drug products with a view to in vitro and in silico simulations: a PEARRL review

Objectives: Using the type of meal and dosing conditions suggested by regulatory agencies as a basis, this review has two specific objectives: first, to summarize our understanding on the impact of food intake on luminal environment and drug product performance and second, to summarize the usefulness and limitations of available in vitro and in silico methodologies for the evaluation of drug product performance after food intake. Key findings: Characterization of the luminal environment and studies evaluating product performance in the lumen, under conditions suggested by regulatory agencies for simulating the fed state, are limited. Various in vitro methodologies have been proposed for evaluating drug product performance in the fed state, but systematic validation is lacking. Physiologically based pharmacokinetic (PBPK) modelling approaches require the use of in vitro biorelevant data and, to date, have been used primarily for investigating the mechanisms via which an already observed food effect is mediated. Summary: Better understanding of the impact of changes induced by the meal administration conditions suggested by regulatory agencies on the luminal fate of the drug product is needed. Relevant information will be useful for optimizing the in vitro test methods and increasing the usefulness of PBPK modelling methodologies

In Vitro Simulation of the Environment in the Upper Gastrointestinal Lumen After Drug Administration in the Fed State Using the TIM-1 System and Comparison With Luminal Data in Adults

We evaluated the environment in TIM-1 luminal compartments using paracetamol and danazol solutions and suspensions and the fed state configuration. Data were compared with recently published data in healthy adults. TIM-1 experiments were performed with a 3-fold downscale. Volumes of secretions in gastric and duodenal compartments adequately reflected the luminal data in adults up to 3 h post drug dosing. pH values in duodenal and jejunal compartments adequately reflected average pH values in adults. In gastric compartment pH values where initially higher than average values in adults and reached baseline levels earlier than in adults. The environment in the TIM-1 gastric compartment and jejunal compartment adequately reflected the average total paracetamol and danazol amounts per volume of contents in the adult stomach and upper small intestine, respectively. Total bile acids concentrations in the micellar phase of contents in duodenal and jejunal compartments overestimated micellar concentrations in the upper small intestine of adults. Adjustments in gastric emptying/acid secretion rates and bile acids identities in the duodenal and jejunal compartments, and application of dynamic bile acids secretion rates are expected to further improve the relevance of luminal conditions in TIM-1 compartments with those in adults. © 2021 American Pharmacists Associatio

Oral biopharmaceutics tools: recent progress from partnership through the Pharmaceutical Education and Research with Regulatory Links collaboration

Objectives: To summarise key contributions of the Pharmaceutical Education and Research with Regulatory Links (PEARRL) project (2016–2020) to the optimisation of existing and the development of new biopharmaceutics tools for evaluating the in vivo performance of oral drug products during the development of new drugs and at the regulatory level. Key findings: Optimised biopharmaceutics tools: Based on new clinical data, the composition of biorelevant media for simulating the fed state conditions in the stomach was simplified. Strategies on how to incorporate biorelevant in vitro data of bio-enabling drug products into physiologically based pharmacokinetic (PBPK) modelling were proposed. Novel in vitro biopharmaceutics tools: Small-scale two-stage biphasic dissolution and dissolution-permeation setups were developed to facilitate understanding of the supersaturation effects and precipitation risks of orally administered drugs. A porcine fasted state simulated intestinal fluid was developed to improve predictions and interpretation of preclinical results using in vitro dissolution studies. Based on new clinical data, recommendations on the design of in vitro methodologies for evaluating the GI drug transfer process in the fed state were suggested. The optimized design of in vivo studies for investigating food effects: A food effect study protocol in the pig model was established which successfully predicted the food-dependent bioavailability of two model compounds. The effect of simulated infant fed state conditions in healthy adults on the oral absorption of model drugs was evaluated versus the fasted state and the fed state conditions, as defined by regulatory agencies for adults. Using PBPK modelling, the extrapolated fasted and infant fed conditions data appeared to be more useful to describe early drug exposure in infants, while extrapolation of data collected under fed state conditions, as defined by regulators for adults, failed to capture in vivo infant drug absorption. Summary: Substantial progress has been made in developing an advanced suite of biopharmaceutics tools for streamlining drug formulation screening and supporting regulatory applications. These advances in biopharmaceutics were achieved through networking opportunities and research collaborations provided under the H2020 funded PEARRL project