Method for simultaneous measurement of norepinephrine, 3-methoxy-4-hydroxyphenylglycol and 3,4-dihydroxyphenylglycol by liquid chromatography with electrochemical detection: application in rat cerebral cortex and plasma after lithium chloride treatment.
An assay was developed to quantify norepinephrine (NE) and its metabolites (MHPG and DHPG) by high-performance liquid chromatography with electrochemical detection method (HPLC-ECD) in brain tissue and plasma of rats treated by LiCl. Separation on C(18) column was obtained by a mobile phase consisting of 4.5% methanol in buffer (0.1 M sodium acetate, 0.2 M citric acid) containing 0.2 mM ethylenediaminetetraacetic acid disodium salt (EDTA Na(2)) and 0.4 mM sodium octylsulfate, operated at a flow rate of 0.8 ml/min. A potential of +0.78 V was applied across the working and reference electrodes of the detector. The precision was in the range 2.88-4.35% for NE, 5.94-11.0% for MHPG and 1.97-4.40% for DHPG. Accuracy was 98.8-99.3% for NE, 97.4-100% for MHPG and 96.1-101% for DHPG. The limit of detection was 0.6 ng/ml for NE, 0.5 ng/ml for MHPG and 0.2 ng/ml for DHPG. The linearity is over the range 20-60 ng/ml for NE, 7-23 ng/ml for MHPG and 6-20 ng/ml for DHPG. The assay has been applied successfully to measure simultaneously cortex and plasmas concentrations of these three catecholamines in rats.
Sastre
Elisabeth
E
Laboratoire de Chimie Analytique, Faculté de Pharmacie, 27 Boulevard Jean Moulin, 13385 Cedex 5, Marseille, France. elisabeth.sastre@pharmacie.univ-mrs.fr
Nicolay
Alain
A
Bruguerolle
Bernard
B
Portugal
Henri
H
D000818:Animals; D002540:Cerebral Cortex / Q000737:chemistry*; D005260:Female; D007700:Kinetics; D018021:Lithium Chloride / Q000008:administration & dosage*; D008734:Methoxyhydroxyphenylglycol / Q000031:analogs & derivatives* / Q000032:analysis* / Q000097:blood; D009638:Norepinephrine / Q000032:analysis* / Q000097:blood; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D008734:Methoxyhydroxyphenylglycol; D018021:Lithium Chloride; C010117:3,4-dihydroxyphenylglycol; D009638:Norepinephrine