Determination of Some Beta Adrenergic Agonists Using Different Spectroscopic and Electrochemical Techniques A Thesis Presented by Marwa Mohamed Abdelmonem Sakr B. Pharm. Sci. (2002) Faculty of Pharmacy, Helwan University M. Pharm. Sci. (2010) Faculty of Pharmacy and Biotechnology, German University in Cairo Submitted for The Degree of Doctor of Philosophy in Pharmaceutical Sciences (Pharmaceutical Chemistry) Supervised by Professor Dr. Samir Mohamed El-Moghazy Professor of Pharmaceutical Chemistry Faculty of Pharmacy Cairo University Professor Dr.Hala Sultan Al-Easa Professor of Organic Chemistry Faculty of Arts and Sciences Qatar University Dr.Marwa Ahmed Fouad Associate Professor of Pharmaceutical Chemistry Faculty of Pharmacy Cairo University Dr.Rasha Sayed Hanafi Associate Professor of Pharmaceutical Chemistry Faculty of Pharmacy and Biotechnology German University in Cairo Faculty of Pharmacy Cairo University 2018 Abstract CHAPTER ONE I. INTRODUCTION In this section, a detailed description of three fluoroquinolones and their types as well as their pharmacological mode of action is defined.. CHAPTER TWO II. LITTERATURE SURVEY It delineates the chemical and physical characteristics, pharmacological actions as well as the techniques of analysis that were previously reported for the determination of norfloxacin, levofloxacin and lomefloxacin. CHAPTER THREE III. EXPERIMENTAL, RESULTS, DISCUSSIONS AND CONCLUSIVE REMARKS III.I. Spectrophotometry III.I.A. Spectrophotometry Experimental It encompasses the materials, reagents, instruments, software used as well as the experimental techniques applied. III.I.B. Spectrophotometry Results and Discussions The interaction of the three fluoroquinlones with lanthanides based on the antenna effect theory was verified and spectrofluorimetrically. III.I.C. Spectrophotometry Conclusive Remarks It entitles a general outline about results obtained in the spectrofluorimetry section. III.II. Spectrofluorimetry III.II.A. Spectrofluorimetry Experimental It includes the materials, reagents, instruments, software used as well as the experimental techniques instigated. III.II.B. Spectrofluorimetry Results and Discussions This section provides a comprehensive enlightenment about the obtained chemometric determination of the spectrophotometrically analyzed complexes. III.II.C. Spectrofluorimetry Conclusive Remarks It provides a general conclusion on the results obtained in the spectrophotometric section. III.III. Potentiometry III.III.A. Potentiometry experimental It presents the materials, reagents and instruments as well as the experimental techniques applied for the determination of norfloxacin, levofloxacin and lomefloxacin III.III.B. Potentiometry Results and discussion Electrodes were prepared and investigated for norfloxacin, levofloxacin and lomefloxacin based on the incorporation of the Tetraphenyl borate (TPB) as an ion- exchanger in PVC matrix using DOP as plasticizer III.III.C. Potentiometry Conclusive Remarks This section ends with a conclusion on the obtained potentiometric results. The thesis ends with 416 references that were of great help to the authoress. INTRODUCTION Adrenergic drug agonists Adrenergic drugs are crucial sympathomimetic agents that serve as analogues of epinephrine (adrenaline), norepinephrine (noradrenaline) and dopamine hormones secreted by the adrenal gland. They resemble these hormones not only in terms of pharmacological actions but also due to their possession of the vital catecholamine functional group. The term adrenergic literally means closely associated with adrenaline (epinephrine) and/or noradrenaline (norepinephrine) . The principle role of the stated hormones is to adapt the body to stressful conditions . When circulating in the blood, their half-life lasts for few minutes, after which they are metabolized either by catechol-O-methyl transferases or by monoamine oxidase .In other words, the pharmacological impacts exhibited by adrenergic drug agonists contributes to the effectual activation of adrenergic receptors. Adrenergic receptors are divided into two categories α and β (α1, α2, β1, and β2 receptors) by which adrenergic drugs exert their pivotal effects. α –receptors are mainly activated by norepinephrine while β receptors are primarily activated by epinephrine, however epinephrine may also activate α receptors. Adrenergic drug agonists are divided into three classes: direct acting, indirect acting, and dual acting . I.II. Classification of β – Adrenergic drug agonists I.II.A. β1-Receptor activating drugs Due to their prevalence in the heart muscle and kidney, β1- are largely associated with the conducting system (e.g., pacemaker) and the ventricular musculature. They are also abundant in sweat and salivary glands. Epinephrine or norepinephrine cause excitatory responses in these tissues. Activation of β1 receptors in the heart has a positive inotropic effect leading to an increase in the force of contraction, thus improving cardiac performance. I.II.B. β2 –Receptor activating drugs β2 Receptors are mostly present in bronchial smooth muscle and blood vessels of skeletal muscle. They can also be found in arterioles, heart, lung, uterus, and liver. Therapeutic applications of β2 stimulation are restricted to the lungs and the uterus. The key role of β2-adrenergic drug agonists, is to cause smooth muscle relaxation in the lung promoting bronchodilation , thus relieving or preventing asthma attacks. Accordingly, these drugs serve as suitable candidates for the treatment of bronchial asthma, chronic bronchitis, emphysema and in cases of premature labor for relaxing uterine smooth muscle. An example of a β2-receptor activating drug is hexoprenaline Trivalent Lanthanides and Lanthanide-catecholamine complexes Owing to their extraordinarily high sensitivity and selectivity , as reported by literature, lanthanide III metals were known to play fundamental roles as luminescent probes in many pharmaceutical and bioanalytical applications. Lanthanide-sensitized luminescence present wide Stokes shift , narrow emission bands along with extended luminescence lifetimes . Owing to their strong emission peaks (<10 nm full width at half-maximum) in the visible and near-infrared region under UV excitation , lanthanide complexes’ luminescence has gained a lot of attention in the past decade . Attributable to their photo-transfiguration properties, these complexes are also pertinent in many other diverse fields . Aim of work Applying a multivariate approach, a chemometric-assisted spectrofluorimetric and spectrophotometric techniques were executed for the determination of Dobutamine (DOB), Hexoprenaline (HEX) and Isoprenaline (ISO) catecholamine drugs in their pure and pharmaceutical dosage forms. Predicated on the existent “antenna” interaction between the catecholamine drugs and the designated lanthanide metals, nine new chemosensors were modeled for the analysis. Lanthanum, Samarium and Terbium metals were selected for the attainment of the designated models through their complexation reactions with DOB, HEX and ISO. Four continuous factors were screened namely, reaction time (RT), metal volume (MV), temperature (T) and pH by aid of Plackett-Burman Design. Optimization for the screened factors was carried out by means of Box-Behnken Design and Central Composite Design for spectrofluorimetric and spectrophotometric techniques, respectively. The present work also aimed to offer three new ion-selective electrodes of the plastic membrane type for the determination of dobutamine hydrochloride (DOB), hexoprenaline sulphate (HEX) and isoprenaline hydrochloride (ISO) in pure and pharmaceutical dosage forms. These electrodes depend mainly on the incorporation of the ion- exchangers of the above mentioned drugs with phosphomolybdic acid (PMA) in a PVC matrix. Electrodes’ performance characteristics were tested by investigating the effects of membrane composition, soaking, temperature and pH. Selectivity of the three electrodes towards their respective ions was also examined in presence of various cations and sugars. Review Spectrofluorimetric Analysis Spectral characteristics Using the spectrofluorimetric technique, maximum fluorescence intensity was detected for the formed complexes. Ln complexes were produced at λex/λem= 312 /455 for DOB-Ln, 310/455 for HEX-Ln and 315/455 for ISO-Ln. For Sm, complexes were obtained at λex/λem= 312 /450 for DOB-Sm, 310/450 for HEX-Sm and 315/450 for ISO-Sm. Tb complexes were produced at λex/λem= 312 /550 for DOB-Tb, 310/550 for HEX-Tb and 315/550 for ISO-Tb. Thus, signifying the antenna effect phenomena through interaction between the catecholamine ligands and the three lanthanides [396]. In order to avoid direct excitation of lanthanide metals, it is worth noting that excitation peaks were carefully chosen at these given wavelengths. Figs. 6-8 demonstrates the excitation and emission spectra for the three blank catecholamines and their spectra after complexation with Ln3+, Sm3+, Tb3+, respectively. Experimental Designs Screening step using Plackett-Burman Design (PBD) In the current work, four chief factors were studied by using a Plackett-Burman Design (PBD) with the aim of constructing the design and screening a set of experiments [87].The design included two levels for each factor with their upper (+) and lower (-) limits being adjusted. Examination of the four factors, namely reaction time (RT), metal volume (MV), temperature (T) and pH for DOB, HEX and ISO lanthanide complexes was carried out and the resultant responses were measured