TY - JOUR
T1 - Determination of free and total (free plus protein-bound) melatonin in plasma and cerebrospinal fluid by high-performance liquid chromatography with fluorescence detection
AU - Rizzo, Vittoria
AU - Porta, Camillo
AU - Moroni, Mauro
AU - Scoglio, Enrico
AU - Moratti, Remigio
PY - 2002/7/5
Y1 - 2002/7/5
N2 - A simple, sensitive and accurate method for the estimation of free and total (free plus protein-bound) melatonin (MLT) in human plasma and cerebrospinal fluid (CSF) is described. Via Chem-Elut cartridges, free and total MLT (the latter obtained after a deproteinization step) were quantified in dichloromethane-extracted samples and analyzed in one chromatographic run by high-performance liquid chromatography (HPLC) with fluorimetric detection. The column used was an Extrasil ODS-2 (3 μm, 150×4.6 mm I.D.), while the mobile phase consisted of 75 mM sodium acetate-acetonitrile (72:28, v/v) (pH 5.0). Repeatability and reproducibility of the method were 3.24 and 9.4%, respectively. The recovery of melatonin from plasma and CSF was 99.9±4.0% for non-deproteinized samples and 93.2±4.8% for deproteinized samples. The detection limit of the assay was 0.5 pg/ml. In human plasma, the mean±SD concentrations in the darkness period were 23.18±7.44 pg/ml for free melatonin and 82.5±36.48 pg/ml for total melatonin, while the lowest concentrations detected during daytime were 2.23±2.22 and 7.40±5.68 pg/ml, respectively. Detection of MLT in CSF was 5.01±2.31 and 28.55±6.95 pg/ml for the free and total fraction, respectively.
AB - A simple, sensitive and accurate method for the estimation of free and total (free plus protein-bound) melatonin (MLT) in human plasma and cerebrospinal fluid (CSF) is described. Via Chem-Elut cartridges, free and total MLT (the latter obtained after a deproteinization step) were quantified in dichloromethane-extracted samples and analyzed in one chromatographic run by high-performance liquid chromatography (HPLC) with fluorimetric detection. The column used was an Extrasil ODS-2 (3 μm, 150×4.6 mm I.D.), while the mobile phase consisted of 75 mM sodium acetate-acetonitrile (72:28, v/v) (pH 5.0). Repeatability and reproducibility of the method were 3.24 and 9.4%, respectively. The recovery of melatonin from plasma and CSF was 99.9±4.0% for non-deproteinized samples and 93.2±4.8% for deproteinized samples. The detection limit of the assay was 0.5 pg/ml. In human plasma, the mean±SD concentrations in the darkness period were 23.18±7.44 pg/ml for free melatonin and 82.5±36.48 pg/ml for total melatonin, while the lowest concentrations detected during daytime were 2.23±2.22 and 7.40±5.68 pg/ml, respectively. Detection of MLT in CSF was 5.01±2.31 and 28.55±6.95 pg/ml for the free and total fraction, respectively.
KW - Melatonin
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U2 - 10.1016/S1570-0232(02)00168-X
DO - 10.1016/S1570-0232(02)00168-X
M3 - Article
C2 - 12052718
AN - SCOPUS:0037025020
VL - 774
SP - 17
EP - 24
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
SN - 1570-0232
IS - 1
ER -