Isolation of melatonin by immunoaffinity chromatography.
A single-step, highly specific and easy-to-use method was developed for isolation and purification of melatonin from complex biological matrices. Polyclonal antibodies highly specific against melatonin (with cross-reactivities with related compounds below 0.02%, except for 6-hydroxymelatonin) were raised, characterised by enzyme-linked immunosorbent assay (ELISA) and used for preparation of immunoaffinity gel. Melatonin recovery by the immunoaffinity method was approximately 95%, allowing single-step processing of samples prior to electrospray HPLC-MS analysis (with detection limit 10 fmol). The method was successfully used for determining melatonin in human serum and turned out to be better than the non-specific solid-phase extraction published earlier.
Rolcík
Jakub
J
Laboratory of Growth Regulators, Palacký University and Institute of Experimental Botany ASCR, Slechtitelů 11, 783 71, Olomouc, Czech Republic. rolcikj@aix.upol.cz
Lenobel
René
R
Siglerová
Vĕra
V
Strnad
Miroslav
M
D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D008550:Melatonin / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization
D008550:Melatonin