An improved approach for the determination of plasma-free catecholamines by HPLC-ED: Application to normal and hypertensive patients |
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Authors: | C Lucarelli P Betto G Ricciarello M Giambenedetti F Sciarra C Tosti Croce P L Mottironi |
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Institution: | (1) Instituto Superiore di Sanità, V. Ie Regina Elena, 299, 00161 Roma, Italy;(2) Va Clinica Medica, Università di Roma, 00161 Roma, Italy |
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Abstract: | Summary A dual-step procedure for the rapid, quantitative isolation of free catecholamines (norepinephrine, epinephrine and dopamine)
from plasma, using a little column of CM-Sephadex and alumina adsorption, is described. Sensitive high performance liquid
chromatography is also discussed, employing an amperometric detector for the quantitative determination. The recovery of the
three catecholamines, and of N-methyldopamine used as the internal standard, was about 70–80%; the detection limits were 2pg
for norepinephrine, 3pg for epinephrine and 3pg for dopamine. The combination of the rather specific and easy to handle two-step
sample clean-up procedure, the high resolving power of the chromatography and the high sensitivity of electrochemical detection
provided a simple method for the determination of free catecholamines in plasma samples of normal and essential hypertensive
subjects under different conditions (supine position for 45 min, standing for 5 and 10 min). It was found that a significant
increase in epinephrine levels (P<0.01) occurred in hypertensive patients under the three conditions studied. |
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Keywords: | Column liquid chromatography Plasma catecholamines Dual-step prepurification process Electrochemical detection Essential hypertension Physical stress |
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