Glucuronide and sulfate catecholamine conjugates in rat and human plasma

Abstract

We have developed a method to selectively estimate free, glucuronidated and sulfated catecholamines (epinephrine [E], norepinephrine [NE], dopamine [DA]) in a single plasma sample. The method incorporates the first step of the catecholamine radioenzymatic assay and the selective enzymatic hydrolysis of conjugates by glucuronidase or sulfatase preparations. The method has been applied to rat and human plasma with a view to determine the relative importance of either conjugate (sulfate or glucuronide) toward free catecholamines. No previous reports were available for the concentration of either conjugate in rat plasma or the level of glucuronide conjugate in human plasma. Both sulfate and glucuronide conjugate of the three catecholamines were found in rat and human plasma, at different levels. Sulfate conjugates predominated in man and glucuronides in rat. In human, hand immersion in ice water for three minutes, which increased free catecholamine levels in the first minutes of the test, elicited too a delayed increase of glucuronide levels at the 30th minute (except for DA glucuronide which was already elevated at the third minute). As to the sulfates, only E sulfate was increased at the 10th minute. Our results suggest that glucuronidation may be an important pathway for catecholamine metabolism in man at rest or under sympathetic stimulation. In rat, our data point out the influence of blood sampling conditions (dietary, catheterization, decapitation) on the studied compounds and suggest that the nearest conditions from basal state may be fulfilled in sucrose-fed catheterized rats. The predominance of glucuronides in rat plasma agrees with previous metabolic reports.