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Catecholamine measurements As described before 15 , UPLC-MS/MS was used to analyze urinary levels of free biogenic monoamines (including catecholamines), and their metabolites were measured by 5 L of urine samples from frozen biobanked (-80C) random samples were thawed on ice and mixed with 50 L of deuterated internal standards (epinephrine-d6, norepinephrine-d6, dopamine-d4, metanephrine-d3, normetanephrine-d3, 4-hydroxy-3-methoxymandelic acid d3, 4-hydroxy-3-methoxyphenyl-d3-acetic-d2 acid, 5-hydroxyindole-4,6,7-d3-3-acetic-d2 acid) and 195 L of 0.2% formic acid in a 2 mL amber UPLC sample vial and the samples were analyzed on a Xevo TQ-S micro quadrupole mass spectrometer with an ESI ionization source, interfaced with Waters Acquity Class-H UPLC equipped with a quaternary pump system (Waters, MA)

Hanna, Department of Medicine, Nineveh college of medicine Biochemistry, Mosul, IRAQ Email: [email protected] INTRODUCTION Cigarette-smoking is a well-known risk factor for atherosclerosis development and its complications including cerebral and cardiovascular diseases(CVD)1,2 through vascular endothelial damage3 that possibly occurs through oxygen free radicals production as superoxide radicals, hydrogen peroxide and hydroxyl radicals.4,5 Several enzymes capable of producing oxygen free radicals including xanthine oxidase, NADPH oxidase, myeloperoxidase, and endotoxin.4,5 As cigarette smoke contains superoxide and reactive nitrogen species that readily react with various biomolecules,1,6-9 it has been hypothesized that some of the adverse effects of smoking may result from oxidative damage to endothelial cells, which results in nitric oxide (NO) shortage.1,10,11 (NO) shortage regulates vascular tone that accelerates insufficiency of coronary artery and vasoconstriction in many different tissues.3,12 Therefore imbalance between oxidants and antioxidants may play an important role in the susceptible smoker.13,14 In addition cigarette smokers have increased inflammatory responses that further enhance their oxidative stress.8,9 Since in humans, uric acid is the most abundant aqueous antioxidant, accounting for up to 60% of serum free radical scavenging capacity15 and is an important intracellular free radical scavenger during metabolic stress including smoking,16,17 therefore , measurement of its serum level reflects the antioxidant capacity.15 The aim of this study is to demonstrate the possible effect of smoking on serum uric acid concentration