by Perpetua E. Tan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1996. / Includes bibliographical references (leaves 149-157). / Abstract --- p.9 / Acknowledgments --- p.12 / List of abbreviations --- p.13 / List of Tables --- p.16 / List of Figures --- p.19 / Chapter CHAPTER 1 --- INTRODUCTION --- p.21 / Chapter CHAPTER 2 --- LITERATURE REVIEWS CATECHOLAMINES: NORADRENALINE AND ADRENALINE --- p.25 / Chapter 2.1 --- History --- p.25 / Chapter 2.2 --- Origin of plasma catecholamines --- p.25 / Chapter 2.3 --- Kinetics of entry and removal --- p.28 / Chapter 2.4 --- Levels present in plasma --- p.30 / Chapter 2.5 --- Some factors affecting plasma CA levels --- p.31 / Chapter 2.5.1 --- Effects of age --- p.31 / Chapter 2.5.2 --- Postural change --- p.32 / Chapter 2.5.3 --- Exercise --- p.32 / Chapter 2.5.4 --- Temperature change --- p.32 / Chapter 2.5.5 --- Stress --- p.33 / Chapter 2.5.6 --- Pregnancy --- p.34 / Chapter 2.5.7 --- Disease --- p.35 / Chapter 2.6 --- Actions in the body --- p.35 / Chapter 2.6.1 --- Plasma endogenous catecholamines --- p.35 / Chapter 2.6.2 --- Plasma exogenous catecholamines and medicine --- p.36 / Chapter 2.6.2.1 --- Clinical uses --- p.36 / Chapter 2.6.2.2 --- Effects --- p.37 / Chapter 2.6.2.3 --- Side effects --- p.38 / Chapter 2.7 --- Binding of catecholamines in plasma --- p.38 / Chapter 2.8 --- Measurement of catecholamines in plasma --- p.38 / Chapter 2.8.1 --- Chemistry --- p.38 / Chapter 2.8.2 --- Extraction and purification --- p.39 / Chapter 2.8.3 --- Biological methods --- p.40 / Chapter 2.8.4 --- Colorimetry --- p.41 / Chapter 2.8.5 --- Radioimmunoassay and radioenzymatic assay --- p.41 / Chapter 2.8.6 --- Enzyme-linked immunoassay --- p.42 / Chapter 2.8.7 --- Gas chromatography --- p.42 / Chapter 2.8.8 --- Liquid chromatography --- p.42 / Chapter 2.8.8.1 --- Fluorometry --- p.43 / Chapter 2.8.8.2 --- Electrochemical detection --- p.43 / Chapter 2.9 --- Plasma protein binding of basic drugs --- p.44 / Chapter 2.9.1 --- Binding to albumin --- p.45 / Chapter 2.9.2 --- Binding to alpha-1-acid-glycoprotein --- p.45 / Chapter 2.9.3 --- Binding to other proteins --- p.45 / Chapter 2.9.4 --- Factors affecting drug binding --- p.46 / Chapter 2.9.4.1 --- Pregnancy --- p.46 / Chapter 2.9.4.2 --- Age --- p.46 / Chapter 2.9.4.3 --- Disease states --- p.46 / Chapter 2.9.5 --- Separation procedures to reveal and follow drug protein binding --- p.47 / Chapter 2.9.5.1 --- Equilibrium dialysis --- p.47 / Chapter 2.9.5.2 --- Ultrafiltration --- p.48 / Chapter 2.9.5.3 --- Ultracentrifugation --- p.48 / Chapter 2.9.5.4 --- Gel Filtration --- p.48 / Chapter CHAPTER 3 --- ANALYTICAL TECHNIQUE : PLASMA CATECHOLAMINE ANALYSIS --- p.49 / Chapter 3.1 --- HPLC determination with coulometric detection of catecholamines --- p.49 / Chapter 3.1.1 --- Introduction --- p.49 / Chapter 3.1.2 --- Basic equipment --- p.49 / Chapter 3.1.3 --- Mobile phase preparation --- p.50 / Chapter 3.1.3.1 --- Reagent A (Citrate-acetate-EDTA buffer) --- p.50 / Chapter 3.1.3.2 --- Reagent B (ion pairing reagent) --- p.50 / Chapter 3.1.3.3 --- Mobile phase mixture --- p.50 / Chapter 3.1.4 --- Detector settings --- p.51 / Chapter 3.1.5 --- Sample collection and storage --- p.51 / Chapter 3.2 --- Reagents and solutions --- p.52 / Chapter 3.2.1 --- Acid-washed alumina --- p.52 / Chapter 3.2.2 --- Tris buffer solution --- p.53 / Chapter 3.2.3 --- Washing solution --- p.53 / Chapter 3.2.4 --- Acetic acid solution --- p.53 / Chapter 3.2.5 --- EDTA-HC1 solution --- p.53 / Chapter 3.2.6 --- Citric acid solution --- p.53 / Chapter 3.2.7 --- Stock solutions --- p.54 / Chapter 3.2.7.1 --- Catecholamine standards --- p.54 / Chapter 3.2.7.2 --- Dihydroxybenzylamine (Internal) standard --- p.54 / Chapter 3.2.8 --- Stripped fresh frozen plasma --- p.54 / Chapter 3.2.9 --- Sorensen's phosphate buffer containing 0.6% NaCl --- p.55 / Chapter 3.2.10 --- Control standards --- p.55 / Chapter 3.3 --- Voltammogram of catecholamines and internal standard used --- p.55 / Chapter 3.4 --- Maintenance of the HPLC-Coulometric detector system --- p.56 / Chapter 3.5 --- Optimization of the extraction method --- p.58 / Chapter 3.5.1 --- Amount of alumina for adsorption of CA --- p.58 / Chapter 3.5.2 --- pH of tris buffer for maximum uptake of CA onto alumina --- p.58 / Chapter 3.5.3 --- Optimum time for maximum uptake of CA onto alumina --- p.59 / Chapter 3.5.4 --- Optimum time for maximum desorption of CA into acid solution --- p.59 / Chapter 3.5.5 --- Optimum volume of acid solution for maximum desorption of CA --- p.60 / Chapter 3.6 --- Validation of the method --- p.60 / Chapter 3.6.1 --- Linearity --- p.60 / Chapter 3.6.2 --- Recovery --- p.61 / Chapter 3.6.3 --- Reproducibility --- p.62 / Chapter 3.6.4 --- Stability --- p.62 / Chapter 3.7 --- Results --- p.63 / Chapter 3.8 --- Discussion --- p.79 / Chapter CHAPTER 4 --- CLINICAL APPLICATIONS OF THE CATECHOLAMINE ASSAY --- p.84 / Chapter 4.1 --- Introduction --- p.84 / Chapter 4.1.1 --- Applications of catecholamines assay in clinical science --- p.84 / Chapter 4.2 --- : PLASMA CATECHOLAMINES AFTER INDUCTION OF ANAESTHESIA AT CAESARIAN SECTION --- p.84 / Chapter 4.2.1 --- Introduction --- p.84 / Chapter 4.2.2 --- Patients and methods --- p.86 / Chapter 4.2.3 --- Blood sampling and storage --- p.87 / Chapter 4.2.4 --- Statistics used --- p.87 / Chapter 4.2.5 --- Results --- p.88 / Chapter 4.2.6 --- Discussion --- p.99 / Chapter 4.3 --- EPINEPHRINE INFILTRATION IN SINUS SURGERY --- p.101 / Chapter 4.3.1 --- Introduction --- p.101 / Chapter 4.3.2 --- Patients and methods --- p.102 / Chapter 4.3.3 --- Blood sampling and storage --- p.103 / Chapter 4.3.4 --- Results --- p.104 / Chapter 4.3.5 --- Discussion --- p.108 / Chapter CHAPTER 5 --- ANALYTICAL TECHNIQUE: PLASMA PROTEIN BINDING OF CATECHOLAMINES --- p.110 / Chapter 5.1 --- Equilibrium dialysis for protein binding of drugs --- p.110 / Chapter 5.1.1 --- Introduction --- p.110 / Chapter 5.1.2 --- Dialyzing apparatus --- p.110 / Chapter 5.1.3 --- Sample collection and storage --- p.111 / Chapter 5.1.4 --- Reagents and solutions --- p.111 / Chapter 5.1.4.1 --- Ascorbic acid --- p.111 / Chapter 5.1.4.2 --- Glutathione --- p.111 / Chapter 5.1.4.3 --- Sodium metabisulfite --- p.111 / Chapter 5.1.4.4 --- Dialysis buffer --- p.111 / Chapter 5.1.5 --- Dialysis membrane --- p.112 / Chapter 5.1.6 --- Equilibrium dialysis --- p.112 / Chapter 5.2 --- Optimization of the binding parameters --- p.113 / Chapter 5.2.1 --- Types of preservatives for stability of catecholamines during dialysis --- p.113 / Chapter 5.2.2 --- Dialysis buffer --- p.114 / Chapter 5.2.3 --- Dialysis time and volume of sample --- p.114 / Chapter 5.2.4 --- Dialysis membrane --- p.115 / Chapter 5.2.5 --- Catecholamines concentration for dialysis --- p.114 / Chapter 5.3 --- Total protein analysis- Lowry Method --- p.115 / Chapter 5.3.1 --- Reagents and solutions --- p.116 / Chapter 5.3.1.1 --- Reagent A (Alkaline copper reagent) --- p.116 / Chapter 5.3.1.2 --- Reagent B (Folin-Ciocalteus phenol reagent with water) --- p.116 / Chapter 5.3.2 --- Stock standard and controls --- p.116 / Chapter 5.3.2.1 --- Human serum albumin standard --- p.116 / Chapter 5.3.2.2 --- Controls --- p.116 / Chapter 5.3.3 --- Procedure --- p.116 / Chapter 5.4 --- Results --- p.117 / Chapter 5.5 --- Discussion --- p.126 / Chapter CHAPTER 6 --- CONCLUSIONS --- p.130 / APPENDIX --- p.134 / CHEMICALS AND REAGENTS --- p.146 / REFERENCES --- p.149